Bacillus

J Am Mosq Control Assoc, 1988 Mar, 4(1), 51 - 6
Mosquito host range and field activity of Bacillus sphaericus isolate 2297 (serotype 25); Lacey LA et al.; The 2297 isolate (serotype 25) of Bacillus sphaericus was bioassayed in the laboratory against 8 species of mosquitoes from 3 subfamilies . The most susceptible species were in the genus Culex and the least susceptible were the Aedes spp . and Toxorhynchites r . rutilus . Primary powders of the 2297 and 2362 (serotype 5a5b) isolates were evaluated in the field in natural and simulated habitats against Culex spp . The larvicidal activity of the two isolates was similar, with longer residual activity observed for both preparations in shaded shallow clear water . Larvicidal activity was curtailed in organically enriched and unshaded habitats . Isolate 2297 provided effective control for at least 1 week in an organically enriched habitat and for over 5 weeks in clear water in a shaded habitat when applied at the rate of 0.25 kg/ha.

Plasmid, 1988 Mar, 19(2), 169 - 73
Cloning and partial characterization of three small cryptic plasmids from Bacillus thuringiensis; Mahillon J et al.; The strain H1.1 of Bacillus thuringiensis var . thuringiensis harbors three small cryptic plasmids: pGI1, pGI2, and pGI3 (8.2, 9.2, and 10.6 kb, respectively) . Two of these plasmids (i.e., pGI2 and pGI3) were successfully cloned in their entirety into the vector pBR322, whereas only overlapping DNA fragments covering pGI1 were obtained in Escherichia coli . A curing-hybridization technique was used to obtain isolates of B . thuringiensis missing one or another small cryptic plasmid . These derivatives were examined for any change in a phenotypic trait, but no specific function could be assigned to one of these plasmids . Hybridization and restriction mapping data revealed that the transposon Tn4430 accounts for 45% of the pGI2 plasmid DNA.

Biofizika, 1988 Mar-Apr, 33(2), 293 - 6
{Liberation of non-penetrating spin labels from erythrocytes and liposomes after treatment with bacterial protein}; Komarov AM et al.; Liberation of non-penetrating spin labels from erythrocytes and liposomes was studied at their interaction with delta-endotoxin of Bacillus thuringiensis israelensis . The yield of non-penetrating spin labels was shown to be a highly sensitive indicator of the disturbance of the membrane barrier function.

Bioorg Khim, 1988 Mar, 14(3), 352 - 8
{Antigenic polysaccharides of bacteria . 31 . The structure of the O-specific polysaccharide chain of Pseudomonas aurantiaca IMB 31 lipopolysaccharide}; Knirel' IuA et al.; The O-specific polysaccharide chain of the Pseudomonas aurantiaca IMV 31 lipopolysaccharide contains N-acetyl-L-fucosamine (FucNAc) and di-N-acetyl-D-bacillosamine (2,4-diacetamido-2,4,6-trideoxyglucose, Bac(NAc)2) in the ratio 2:1 . On the basis of methylation, solvolysis with anhydrous hydrogen fluoride, and computer-assisted analysis of 13C-NMR spectrum, it was concluded that the trisaccharide repeating unit of the polysaccharide possesses the following structure: structure: ----3)-beta-D-Bac(NAc)2-(1----3)-alpha-L-FucNAc-(1----3)-alpha-L- FucNAc-(1----.

Biochem Biophys Res Commun, 1988 Feb 29, 151(1), 25 - 31
Nucleotide sequence of the maltohexaose-producing amylase gene from an alkalophilic Bacillus sp . #707 and structural similarity to liquefying type alpha-amylases; Tsukamoto A et al.; The nucleotide sequence of the gene for maltohexaose-producing amylase from an alkalophilic Bacillus sp . #707 was determined . Starting at an ATG initiation codon, an open reading frame was composed of 1554 bp (518 amino acids) . The NH2-terminal portion encoded a 33 amino acid-long signal peptide . The deduced amino acid sequence of the extracellular mature enzyme was more than 60% homologous to those of the liquefying type alpha-amylases but not to those of the saccharifying type alpha-amylases . The sequence of its signal peptide was completely different from those of other alpha-amylases.

Biochem Biophys Res Commun, 1988 Feb 29, 151(1), 48 - 52
Glucose metabolism via the Embden-Meyerhof pathway is not involved in ATP production during spore germination of bacillus megaterium QM B1551 . A study with a mutant lacking hexokinase; Sano K et al.; In order to investigate contributions by glucose metabolism via the Embden-Meyerhof pathway and that via the direct oxidation route to gluconate to initial ATP production during spore germination, respiratory activity and RNA synthesis were compared between the mutant lacking hexokinase and the parent spores of Bacillus megaterium QM B1551 . We found that time courses of those metabolic events were almost identical between those spores, thus clearly indicating that NADH formed by a spore-specific enzyme glucose dehydrogenase (EC 1.1.1.47) is solely responsible for aerobic production of ATP at this stage.

Am J Ophthalmol, 1988 Feb 15, 105(2), 195 - 7
Bacillus keratitis associated with contaminated contact lens care systems; Donzis PB et al.; We examined two soft contact lens wearers who developed keratitis associated with Bacillus contamination of their contact lens care systems . Patient 1 developed a corneal ulcer caused by B . subtilis, and Patient 2 demonstrated multiple, diffuse, punctate corneal epithelial opacities associated with B . cereus contamination in the contact lens and lens case compartment . The contact lens cases of both patients contained Bacillus spores that survived multiple heat disinfection treatments . Three different contact lens chemical disinfection systems used for the minimum recommended time failed to kill the Bacillus organisms.

Biochim Biophys Acta, 1988 Feb 10, 952(3), 282 - 9
Purification and characterization of NADP+-linked isocitrate dehydrogenase from an alkalophilic Bacillus; Shikata S et al.; We have succeeded in purifying to homogeneity a very labile NADP+-linked isocitrate dehydrogenase (isocitrate: NADP+ oxidoreductase (decarboxylating), EC 1.1.1.42) from a strain of alkalophilic Bacillus, by a simple method, with an overall yield over 76% of the original activity . The molecular weight on Sephadex G-200 was around 90,000; and that by electrophoresis on SDS-polyacrylamide gels was about 44,000 . The sedimentation coefficient (s020,w) and isoelectric point of the enzyme were determined to be 3.22 S and pH 4.7, respectively . The enzyme required Mn2+ for the reaction and for stability . The optimum pH for the reaction was in the range 7.8-8.4 at 30 degrees C; the optimum temperature at pH 8.0 was 75 degrees C; the activation energy of the reaction was 6.2 kcal/mol . The Km values for threo-Ds-isocitrate, DL-isocitrate, and NADP+ were 5.4 microM, 9.9 microM, and 7.3 microM, respectively . This enzyme was inhibited by NADPH, glyceraldehyde 3-phosphate, 3-phosphoglycerate, phosphoenol pyruvate, cis-aconitate, alpha-ketoglutarate, and oxaloacetate . In addition, it was subject to a concerted inhibition by a combination of glyoxylate and oxaloacetate, and also to a cumulative inhibition by nucleoside triphosphates.

Biochemistry, 1988 Feb 9, 27(3), 1041 - 9
Asymmetry of tyrosyl-tRNA synthetase in solution; Ward WH et al.; The tyrosyl-tRNA synthetase from Bacillus stearothermophilus crystallizes as a symmetrical dimer with each subunit having a complete active site . The enzyme-substrate complexes, however, are known to be asymmetrical in solution because the enzyme exhibits half-of-the-sites activity by binding tightly only 1 mol of tyrosine or 1 mol of tyrosyl adenylate per mole of dimer . Evidence is now presented that the unligated enzyme is also asymmetrical in solution . Symmetry was investigated by construction of heterodimers containing one full-length subunit and one truncated subunit, allowing the introduction of different mutations into each monomer . Each dimer is active at only one site, but the site used is randomly distributed between the subunits . Each heterodimer thus consists of two equal populations, one activating tyrosine at a full-length subunit and the other at the truncated subunit . No detectable interconversion is found between active and inactive sites over several minutes either in the absence of substrates or when the enzyme is turning over in the steady state . Kinetic evidence implies that wild-type enzyme is inherently asymmetrical even in the absence of substrate.

J Mol Biol, 1988 Feb 5, 199(3), 525 - 37
Crystal structure of neutral protease from Bacillus cereus refined at 3.0 A resolution and comparison with the homologous but more thermostable enzyme thermolysin; Pauptit RA et al.; Neutral protease from Bacillus cereus exhibits a 73% amino acid sequence homology to thermolysin, for which an accurate crystal structure exists . The B . cereus enzyme is, however, markedly less thermostable . The neutral protease was crystallized and diffraction data to 3.0 A resolution were recorded by oscillation photography . The crystal structure was solved by molecular replacement methods using thermolysin as a trial molecule . The solution was improved by rigid-body refinement and model rebuilding into electron density omit-maps . The atomic co-ordinates were refined to R = 21.7% at 3.0 A resolution . Comparison of the resultant model with the thermolysin structure shows that the two enzymes are very similar with a root-mean-square deviation between equivalent C alpha-atoms of 0.88 A . The gamma-turn found in thermolysin is transformed into a beta-turn in the neutral protease by the insertion of a glycine residue . There appear to be no contributions to the enhanced thermostability of thermolysin from additional salt bridges, whereas contributions in the form of extra hydrogen bonding interactions could be important . Other factors that may affect thermostability include the two glycine to alanine exchanges and perturbations in the environment of the double calcium site.

Pediatr Dermatol, 1988 Feb, 5(1), 1 - 9
Dermatologic manifestations and update of cat scratch disease; Margileth AM; Cat scratch disease is a relatively common cause of chronic (three weeks or longer) lymphadenopathy, with 80% of cases occurring in children and adolescents . This self-limited infection caused by a small, gram-negative, pleomorphic bacillus has been identified in ocular granuloma, skin inoculation lesions, and lymph node specimens . Dermatologic manifestations observed prospectively in 908 patients included primary cat scratch inoculation papules, pustules or rarely, vesicles . Occasionally, enanthematous mucous membrane (oral, ocular) primary inoculation lesions were observed . About 5% of patients have generalized macular, maculopapular, morbilliform, and rarely petechial, usually nonpruritic exanthem . Rarely, erythema nodosum or multiforme and ecchymoses with petechial rashes are seen . Thrombocytopenic purpura is extremely uncommon . Unusual manifestations such as the oculoglandular syndrome of Parinaud, encephalopathy, or severe systemic disease occur in about 10% of patients . Management consists of symptomatic treatment and occasional aspiration of a suppurative node . The disease usually resolves spontaneously in two to four months.

J Biol Response Mod, 1988 Feb, 7(1), 65 - 76
Endogenous production of cytotoxic factors in serum of BCG-primed mice by monophosphoryl lipid A, a detoxified form of endotoxin; Bennett JA et al.; Monophosphoryl lipid A (MPL), a detoxified form of endotoxin, was evaluated for its ability to elicit cytotoxic factors in the serum of mice pretreated with BCG . BDF1 mice were given a priming dose of bacillus Calmette-Guerin (BCG) intravenously (i.v.) . Two weeks later, these mice were challenged with MPL i.v . and their serum was tested for cytotoxicity against Lewis lung carcinoma cells growing in culture . A well-tolerated dose of MPL induced substantial serum cytotoxic activity comparable to that found after a toxic dose of endotoxin . The effective dose of MPL for eliciting serum cytotoxicity was 20 times less than the toxic dose of MPL in BCG-primed mice . No serum cytotoxicity was induced by MPL without prior treatment with BCG or in mice exposed to BCG for less than 10 days . The i.v . route of administration was superior to intraperitoneal, intrapleural, or subcutaneous routes for both BCG priming and induction of serum activity with MPL . Serum manifested TNF-like activity in that it was heat-stable, not species-specific, more effective against tumor than normal cell lines, and more effective against a TNF-sensitive than a TNF-resistant cell line . We conclude that MPL is an effective, well-tolerated biological response modifier that triggers production of cytotoxic factors in serum of mice with an activated reticuloendothelial system.

Biophys Chem, 1988 Feb, 29(1-2), 17 - 29
Approaching the molecular structure of ribosomes; Yonath A et al.; Fifteen forms of three-dimensional crystals and three forms of two-dimensional sheets from ribosomal particles have been grown . In all cases only biologically active particles could be crystallized, the crystalline material retaining its integrity and biological activity for months . Cryastallographic data have been collected from crystals of 50 S ribosomal subunits, using synchrotron radiation, at temperatures between 19 and -180 degree C . Although at around 0 degrees C in the synchrotron X-ray beam the crystals rapidly lose their high-resolution reflections, at cryo-temperatures hardly any radiation damage occurs over long periods, and a complete set of diffraction data to about 6 A resolution could be collected from a single crystal . Heavy-atom clusters were used for soaking as well as for specific binding to the surface of the ribosomal subunits prior to crystallization . The 50 S ribosomal subunits from a mutant of Bacillus stearothermophilus which lacks the ribosomal protein BL11 crystallize isomorphously with the native form . Models of the entire 70 S ribosome and of the 50 S subunit have been reconstructed from two-dimensional sheets at 47 and 30 A, respectively . These models demonstrate the overall shape of the particles, the contact areas between large and small subunits, the space where protein biosynthesis may take place and a tunnel through the 50 S subunit which could provide a path for the nascent polypeptide chain.

Gan To Kagaku Ryoho, 1988 Feb, 15(2), 297 - 300
{Prevention of immunodeficiency induced by cancer chemotherapy with BCG}; Miyata T et al.; MFC (MMC, 5-FU and cytosine-arabinoside) therapy applied with the liver organism Bacillus Calmette-Guerin (BCG) for the treatment of postoperative patients with cancer of the digestive organs presenting at stage II and III . Immunological parameters included skin reaction by purified protein derivative (PPD), lymphatic blastogenesis test using phytohemagglutinin (PHA) and lymphatic subsets . The frequency of MFC therapy was significantly higher in the MFC plus BCG group than in the MFC group (p less than 0.001) . At the completion of MFC therapy, both of PHA blastogenesis rate, OKT4/OKT8 and OKT3 were all within the normal limits . The PPD skin reaction was positive (18.2 +/- 4.0mm) as before the start of MFC therapy . These results suggest that BCG immunotherapy may potentiate the effect of chemotherapy.

J Urol, 1988 Feb, 139(2), 410 - 4
Role of fibronectin in intravesical BCG therapy for superficial bladder cancer; Ratliff TL et al.; Intravesical bacillus Calmette-Guerin (BCG) has been demonstrated to be effective both for prophylaxis and treatment of superficial bladder cancer . In order to identify the progression of events that result in BCG-mediated antitumor activity, studies were performed to evaluate the mechanism of binding of BCG within the bladder . Histological and quantitative studies in a mouse model revealed that BCG attached to the bladder wall only in areas of urothelial damage . Preliminary in vitro data showed that BCG attached to surfaces coated with extracellular matrix proteins . Further studies were then performed using purified extracellular matrix proteins to identify the proteins responsible for attachment . BCG were observed to attach to surfaces coated only with purified fibronectin (FN) but not to other purified proteins including laminin, collagen or fibrinogen . The attachment of BCG to purified FN in vitro was dose dependent and was inhibited by anti-FN antibodies . Moreover, BCG attachment in vivo to bladders with damaged urothelial surfaces was inhibited more than 95% by anti-FN antibodies, but binding was not affected by anti-laminin antibodies or preimmune serum . A survey of commercially available BCG vaccines (Pasteur, Tice, Glaxo, Connaught) showed that only Glaxo BCG did not attach to FN-coated surfaces . Glaxo BCG also was shown to express inferior antitumor activity suggesting that the absence of FN binding by Glaxo may have been associated with the absence of antitumor activity of the vaccine.

J Gen Microbiol, 1988 Feb, 134 ( Pt 2), 531 - 8
The secreted antigens of Mycobacterium tuberculosis and their relationship to those recognized by the available antibodies; Abou-Zeid C et al.; Proteins secreted by strains of Mycobacterium tuberculosis during short-term, zinc-sufficient batch culture were identified in order to define antigens likely to be relevant to the pathogenesis of human disease . {35S}Methionine-labelled proteins in supernatants of 4-7 d cultures were separated by PAGE under both denaturing and non-denaturing conditions, and the position of labelled material was determined . Secreted protein patterns of M . tuberculosis were quite similar to those of Bacillus Calmette-Guerin (BCG) but differed by the absence of the 46 kDa dimeric protein specific to BCG and by the presence in large amounts of a 23 kDa protein which, when denatured, gave 13 kDa subunits . This 13 kDa subunit protein constituted up to 20% of secreted proteins in classical strains of M . tuberculosis of phage type B but was not detected in phage type I strains from South India . This may be relevant to the different pathogenicity of these strains . Western blot analysis showed that antigens defined in supernatants of short-term (3 d) cultures of M . tuberculosis constituted a small subset of those seen in supernatants of organisms cultured for longer periods . One of the secreted proteins has the interesting property of binding to fibronectin . The available monoclonal antibodies and antisera have been used to identify lines on immunoblots corresponding to the secreted/released antigens of M . tuberculosis . The present findings suggest that there are major secreted antigens to which antibodies do not yet appear to have been produced experimentally.

Diagn Microbiol Infect Dis, 1988 Feb, 9(2), 123 - 5
False positive Legionella pneumophila direct immunofluorescent monoclonal antibody test caused by Bacillus cereus spores; Flournoy DJ et al.; Direct immunofluorescent monoclonal antibody stain testing for Legionella pneumophila in Oklahoma lake water yielded an unknown bacillus with fluorescence intensity equal to that of L . pneumophila stock strains . The organism in question was identified as Bacillus cereus, a ubiquitous bacterium . When B . cereus cultures were studied, fluorescence was seen in spores but not in vegetative cells . Since a positive immunofluorescent monoclonal antibody test (alone) might be considered by some individuals as unequivocal to very good evidence for the presence of L . pneumophila, this finding emphasizes the importance of confirming positive stain results with cultures whenever possible.

Genetika, 1988 Feb, 24(2), 210 - 5
{Cloning of the restriction-modification genes of Bacillus centrosporus in Escherichia coli}; Povilenis PI et al.; A genomic library of Bacillus centrosporus was obtained using pBR327 as a vector . The total plasmid DNA of the library was cleaved by the BcnI restriction endonuclease and then transformed in Escherichia coli RR1 . Two clones possessing restriction and DNA modification profiles of BcnI were identified among the transformants . Their respective plasmids were 13.3 and 9.05 kbp in size . Restriction mapping of both plasmids showed each of them to contain two sites for HindIII and one for both Eco31I and Eco47III, located at the same distance . This was assumed to be the location region of the BcnI restriction-modification genes . Confirmation of the assumption was obtained by deletion mapping of the recombinant plasmids . Special features concerning cloning of the restriction-modification genes are discussed on the basis of the results obtained.

Ital J Neurol Sci, 1988 Feb, 9(1), 31 - 4
Neurotoxicity of antituberculous drugs in a patient without active tuberculosis; Leppert D et al.; A 56-year old patient presented 3 months after initiation of an antituberculous regimen with Isoniacid (INH, 5 mg/kg daily), Ethambutol (20 mg/kg daily) and Rifampicin (675 mg daily) a mild sensory polyneuropathy and a bilateral retrobulbar neuritis which progressed to a severe optic atrophy . Multiple hyperintense foci were detected with NMR-imaging in the cerebral white matter suggestive of demyelination . INH and Ethambutol are known for their neurotoxic effects but suggestion was made that neurologic signs may not be due to drug neurotoxicity but could be induced by immunological processes initiated by the tubercle bacillus . In the reported patient the suspected tuberculosis of the urogenital tract was never proved histologically . Most likely his neurological symptoms were therefore cause by the administration of INH and Ethambutol . Patients with a low serum zinc level and a slow acetylation of INH are reported to be at special risk; both factors were present in our patient.

FEBS Lett, 1988 Jan 18, 227(1), 61 - 5
Differential transcription of the bla regulatory region during induction of beta-lactamase in Bacillus licheniformis; Salerno AJ et al.; Induction of beta-lactamase (blaP) in Bacillus licheniformis involves the regulatory genes blaI (repressor), blaR1 (coinducer) and R2 (function unknown) . Transcription of the bla genes during induction was followed by Northern hybridization . In the first 30 min 2.3-kb transcripts encoding blaI and blaR1 were present . Subsequently, blaP mRNA and short transcripts encoding only blaI accumulated and reached a peak at 1 h . All bla transcripts turn over rapidly . Active repressor is not required for the burst of blaI-blaR1 mRNA . The production of blaI-blaR1 mRNA, and thus of BlaR1, is probably controlled both at initiation of transcription and at a later step in its synthesis and degradation.

J Immunol, 1988 Jan 15, 140(2), 494 - 500
Endotoxin-induced interferon-gamma production in culture cells derived from BCG-infected C3H/HeJ mice; Matsumura H et al.; The cultured cells prepared from the spleens and peritoneal exudate cells of the C3H/HeJ strain of mice produce very little or no interferon (IFN) by stimulation of bacterial lipopolysaccharide (LPS) . However, the cells taken from LPS-non-responder C3H/HeJ mice which had been infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG) prior to the experiment were capable of producing IFN in culture in the presence of LPS . The peritoneal exudate cells of BCG-primed C3H/HeJ mice were separated into adherent cell and nonadherent cell populations by their adhesiveness to plastic culture dishes . IFN production required the presence of both these cell populations in the same culture, and the IFN activities produced were mainly IFN-gamma . The cultures with nonadherent cells and fixed adherent cells still produced IFN, but the cell cultures reconstituted with the BCG-primed cell population and unprimed cell population produce little if any IFN-gamma . Moreover, when both of the populations were cultured in Marbrook culture vessels separated by a membrane filter, the cultures produced very little or no IFN-gamma . These results indicate that there is a mechanism of IFN-gamma induction by LPS which requires the direct contact between adherent cells and nonadherent cells without the participation of any soluble factor(s) from the adherent cells . The producer cells were mainly in the nonadherent cell population . Previous treatment of nonadherent cells with anti-Thy-1.2 antibody, anti-Lyt-1.1 antibody, anti-L3T4 antibody, or anti-asialo-GM1 antibody and complement diminished the ability of the cells for LPS-induced IFN production with the help of adherent cells . Therefore, it is concluded that both T cells (presumably L3T4+T cells) and asialo-GM1+ natural killer cells in the BCG-primed C3H/HeJ cell cultures produced IFN-gamma in the presence of LPS, and the production was supported by the function of macrophages.

Biochem Biophys Res Commun, 1988 Jan 15, 150(1), 185 - 91
Polyamine metabolism in an obligately alkalophilic Bacillus alcalophilus that grows at pH 11.0; Chen KY et al.; Bacillus alcalophilus, an obligately alkalophilic bacterium that grows at pH 11.0, has an intracellular pH of 9.5 or less . Unlike all other living organisms, polyamines (putrescine, spermidine and spermine) in B . alcalophilus, if present, will be largely unprotonated . HPLC analysis indicated that spermidine is the major polyamine in B . alcalophilus, accounting for more than 90% of total polyamines, and the level of spermidine varies during growth . Ornithine decarboxylase activity was not detectable in B . alcalophilus under all conditions examined . When {3H}arginine was added to the culture medium, the radioactivity can be recovered from polyamine pool; the distribution is 3% for putrescine, 94% for spermidine, and 3% for spermine, suggesting the presence of arginine pathway for polyamine biosynthesis . The polyamine transport system in B . alcalphilus appears to be Na+-dependent and is highly sensitive to the inhibition of gramicidin S and valinomycin.

Biochim Biophys Acta, 1988 Jan 13, 937(1), 195 - 203
Interrelationships between tyrocidine and gramicidin A' in their interaction with phospholipids in model membranes; Aranda FJ et al.; (1) The interaction of tyrocidine with different lipids is studied in model membranes and the results are compared to the gramicinid-lipid interaction . (2) The tyrocidine-dielaidoylphosphatidylethanolamine interaction gives rise to a population of phospholipids with a lower gel to liquid-crystalline transition temperature and to an abolition of the bilayer to HII phase transition, resulting in a macroscopic organization with dynamic and structural properties different from those of the pure lipid . (3) Tyrocidine has a strong fluidizing effect on the acyl chains of phosphatidylcholines, manifested by a decrease in enthalpy of the main thermotropic transition . (4) No evidence of a gramicidin A'-like lipid-structure modulating activity was found . However, tyrocidine inhibits the formation by gramicidin of an HII phase in dioleoylphosphatidylcholine model membranes . Instead, a cubic type of lipid organization is observed . (5) Tyrocidine greatly perturbs the barrier properties of dioleoylphosphatidylcholine model membrane . (6) Gramicidin A' reverses the effect of tyrocidine on membrane permeability by forming a complex in the model membrane with an apparent 1:1 stoichiometry . (7) The results suggest that both peptide antibiotics, which are produced by Bacillus brevis ATC 8185 prior to sporulation, show antagonism in their effect on membrane structure similar to their effect on superhelical DNA (Bogh, A . and Ristow, H . (1986) Eur . J . Biochem . 160, 587-591 . The possible underlying basic mechanism is indicated.

J Mol Biol, 1988 Jan 5, 199(1), 183 - 93
Tertiary structure of Bacillus thermoproteolyticus {4Fe-4S} ferredoxin . Evolutionary implications for bacterial ferredoxins; Fukuyama K et al.; The structure of a low-potential {4Fe-4S} ferredoxin from Bacillus thermoproteolyticus has been solved using anomalous scattering data from iron atoms in the diffraction data of native crystals and refined partially to a crystallographic R-factor of 0.33, with 2.3 A (1 A = 0.1 nm) resolution data . The least-squares refinement based on the Bijvoet differences has determined that the four iron atoms in the cluster are an equal distance, approximately 2.8 A, apart . The NH .. . S hydrogen bonds between polypeptide nitrogen atoms, and both cysteine and inorganic sulfur atoms, are present, as in ferrodoxin from Peptococcus aerogenes . The polypeptide chain of the B . thermoproteolyticus ferredoxin has a fold closely similar to that of 2{4Fe-4S} ferredoxin from P . aerogenes . The structural correspondence indicates strongly that both types of ferredoxin evolved from a common ancestor . The second cluster-binding region in P . aerogenes ferredoxin corresponds to the alpha-helix in B . thermoproteolyticus ferredoxin . The secondary-structure predictions strongly suggest that the alpha-helix is generally present in the monocluster-type ferredoxins . The conformational change to alpha-helix, insertions of a loop and a protrusion, as well as the absence of the second cluster in B . thermoproteolyticus ferredoxin, result in the lack of 2-fold symmetry present in P . aerogenes ferredoxin . So, the track of gene duplication is no longer detectable in the tertiary structure alone . The evolutionary events that may have occurred in the ferredoxins with the {4Fe-4S} cluster are discussed.

J Biol Chem, 1988 Jan 5, 263(1), 561 - 7
Amino acid sequence and entomocidal activity of the P2 crystal protein . An insect toxin from Bacillus thuringiensis var . kurstaki; Donovan WP et al.; The gene encoding the 66-kDa entomocidal protein (P2 protein or mosquito factor) from Bacillus thuringiensis var . kurstaki has been isolated by the use of a 62-mer oligonucleotide probe that encoded 21 amino acids of the P2 protein NH2 terminus . The DNA sequence of the gene, designated cryBI, was unique from the published sequences of other B . thuringiensis genes . However, the amino acid sequence of the P2 protein, as deduced from the DNA sequence of the cryBI gene, was found to contain a sequence of 100 amino acids having 37% homology to a group of B . thuringiensis entomocidal proteins, the P1 proteins . Late stationary phase Bacillus megaterium cells harboring the cloned B . thuringiensis cryBI gene contained large aggregates of the P2 protein, and the cells were highly toxic to both lepidopteran and dipteran larvae . In contrast, Escherichia coli cells harboring the cloned cryBI gene contained very low levels of the P2 protein . DNA blot hybridization experiments showed that certain B . thuringiensis strains contained at least one cryBI-related DNA sequence in addition to the cryBI gene itself.

Eur J Biochem, 1988 Jan 4, 170(3), 507 - 14
Nucleotide sequence and linkage map position of the genes for ribosomal proteins L14 and S8 in the maize chloroplast genome; Markmann-Mulisch U et al.; The nucleotide sequence of a 1287-base-pair segment of the maize (Zea mays) chloroplast DNA, encoding chloroplast ribosomal proteins L14, S8 and the C-terminal part of L16, has been determined using the dideoxy-chain-termination method . These data from a monocot plant are compared to the corresponding data from a dicot and a lower plant and from two bacteria . The deduced amino acid sequence of maize chloroplast L14 shows 80%, 81%, 51% and 52% and that of S8 shows 75%, 58%, 39% and 38% sequence identity, respectively, to the corresponding sequences of Nicotiana tabacum, Marchantia polymorpha, Bacillus stearothermophilus and Escherichia coli . The starting map coordinates of rpL14 and rpS8 in the physical map of the maize chloroplast DNA {Larrinua, I . M., Muskavitch, K . M . T., Gubbins, E . J . and Bogorad, L . (1983) Plant Mol . Biol . 2, 129-140} are 31.330 and 31.841 . The gene order is rpL16-spacer-rpL14-spacer-rpS8 . Shine-Dalgarno sequences (GGA and AGGAGG) and computer-derived stem-loop structures of dyad symmetry are present in the spacers and the 3' downstream region of rpS8, respectively, but a chloroplast promoter-like sequence could not be detected suggesting that the latter might be located further upstream in this ribosomal protein gene cluster in maize chloroplast DNA.

Antonie Van Leeuwenhoek, 1988, 54(3), 257 - 65
Affinity purification of a 65-kilodalton parasporal protein from Bacillus thuringiensis PG-14 that shows mosquitocidal activity; Yu YM et al.; By using antibody-mediated affinity chromatography, a highly mosquito larvicidal but nonhemolytic fraction was obtained from alkali-solubilized, silkworm (Bombyx mori) larval gut juice-treated parasporal inclusions of Bacillus thuringiensis strain PG-14 (serotype 8a:8b) . This fraction contained a 65-kDa protein only but not a 25-kDa protein, the main component in the flow through fraction unbound to the affinity column . The 25-kDa protein purified from the unbound fraction by CM-cellulose chromatography demonstrated a high hemolytic activity against sheep red blood cells but very low mosquito larvicidal activity.

Arch Microbiol, 1988, 149(6), 485 - 91
Localized insertion of new S-layer during growth of Bacillus stearothermophilus strains; Gruber K et al.; Bacillus stearothermophilus strains PV 72 and ATCC 12980 carry a crystalline surface layer (S-layer) with hexagonal (p6) and oblique (p2) symmetry, respectively . Sites of insertions of new subunits into the regular lattice during cell growth have been determined by the indirect fluorescent antibody technique and the protein A/colloidal gold technique . During S-layer growth on both bacillus strains the following common features were noted: 1 . shedding of intact S-layer or turnover of individual subunits was not seen; 2 . new S-layer was deposited in helically-arranged bands over the cylindrical surface of the cell at a pitch angle related to the orientation of the lattice vectors of the crystalline array; 3 . little or no S-layer was inserted into pre-existing S-layer at the poles, and 4 . septal regions and, subsequently, newly formed cell poles were covered with new S-layer protein.

Jikken Dobutsu, 1988 Jan, 37(1), 67 - 72
Enzyme-linked immunosorbent assay for detection of serum antibody to CAR bacillus; Shoji Y et al.; An enzyme-linked immunosorbent assay (ELISA) for detection of CAR bacillus antibody in rat sera was developed by Ganaway et al., in 1985 although the ELISA method was not described in detail . We investigated antigen preparation and test procedures of the ELISA using two strains of CAR bacillus which we isolated from a mouse (CB-M) and a rat (CB-R) . Allantoic fluids containing 2.4 X 10(8)/ml of CB-M and 2.0 X 10(8)/ml of CB-R were washed with sterile phosphate buffered saline (PBS), resuspended in a 1/5 volume of sterile carbonate buffer (pH 9.8) and sonicated . Then 1/40 and 1/80 dilutions of CB-M and CB-R lysates in PBS, respectively, were used for antigen solutions of ELISA . Briefly, antibodies in sera are reacted with antigens coated on the surface of microtiter plates . The amount of horse radish peroxidase labeled protein-A or anti-rat IgG bound to the antigen-antibody complexes is measured on the spectro photometer at wave length of 492 nm . A total of 180 mouse and 205 rat sera were tested against both antigens . The optical density (OD) values of 140 mouse and 161 rat sera obtained from SPF mice and rats free from CAR bacillus infection were on the average 0.005 and 0.019, respectively . On the other hand, OD values of the sera collected from CB-M or CB-R infected animals ranged from 0.20 to 1.52 . According to these results, the cut-off OD value for positive reaction was set at 0.1.(ABSTRACT TRUNCATED AT 250 WORDS)

J Reprod Med, 1988 Jan, 33(1), 41 - 5
Bacillus Calmette-Guérin immunotherapy in ovarian cancer; Pattillo RA et al.; Forty-six advanced ovarian cancer patients treated with conventional modalities with the addition of bacillus Calmette-Guerin (BCG) immunotherapy showed prolonged survival when compared to controls not given BCG . Although the data suggest enhancement of survival with the addition of BCG to conventional treatment, the fact remains that disease recurrence ultimately claims the lives of most of these patients . Nonetheless, patients are surviving longer in the face of advanced disease.

Ann Fr Anesth Reanim, 1988, 7(1), 76 - 80
{Surface changes of intravenous catheters after antineoplastic chemotherapy}; Ranchere JY et al.; During long-term venous catheter implantation, septic and thrombotic complications are quite frequent . In the case reported, the failure of systemic and local antibiotic therapy during repeated septicaemia due to Bacillus cereus at the time of intensive chemotherapy led to a scanning electron microscopy study of the used silicone catheter . There were marked changes of the inner surface with a lot of cellular remains, in contrast with the usual non thrombogenic property of the silicone . An in vitro study was carried out with antitumour agents . Duration of exposure and drug concentration were identical to those used in in vivo perfusions . There were marked changes of the inner surface, which could lead to important modifications of the properties of the silicone . The damage depended on the drug . Silicone was slightly sensitive to vicristin and carmustin, but highly sensitive to cisplatin and doxorubicin . The compatibility of catheter material with the drugs used, especially for oncologic chemotherapy, must be tested systematically.

J Trauma, 1988 Jan, 28(1 Suppl), S222 - 5
Effects of phospholipase C, a tissue thromboplastin inhibitor, on pulmonary microembolism after missile injury of the limb; Jansson IG et al.; Tissue thromboplastin probably plays an important role in the development of post-traumatic pulmonary microembolism . Infusion of purified human tissue thromboplastin in animals resulted in an intravascular coagulation and respiratory insufficiency . This could be inhibited by previous infusion of phospholipase C (PLC) from Bacillus cereus . We have studied the effects of PLC infusion on the course of post-traumatic pulmonary microembolism, induced by a high-energy (c . 700 J) missile trauma to the hind legs of pigs . The trauma resulted in a major muscular injury and an indirect femoral fracture . Untreated pigs developed intrapulmonary microemboli . The degree of microembolism in the lungs was measured quantitatively by external detection over the right lung of radiolabeled platelets and fibrin . Infusion of 80 micrograms PLC/kg/hour resulted in an accumulation of blood PLC associated with toxic reaction leading to increasing tachycardia and circulatory collapse after 10 hours . PLC infusion of 20 micrograms/kg/hour did not inhibit the pulmonary microembolism . A PLC-dose in between, viz . 40-50 micrograms/kg/hour, proved to efficiently inhibit most of the microembolism during the infusion period . Cessation of PLC infusion after 24 hours was accompanied by a later increase in pulmonary trapping of platelets and fibrin and decreases in paO2 . Concomitantly there were opacities seen on chest X-rays . The results show that tissue thromboplastin is an important etiologic factor in post-traumatic pulmonary microembolism and that inhibition with phospholipase C can be of value in the prophylaxis of the syndrome.

Drugs, 1988, 35 Suppl 2, 185 - 9
Cefotaxime treatment of gram-negative enteric meningitis in infants and children; Jacobs RF; 18 infants and children (1 week to 3 months of age) were treated with cefotaxime 200 mg/kg/day for Gram-negative enteric bacillary meningitis . 17 of these patients (94.4%) survived, with a complication rate of 23.5% (4/17 patients) . The follow-up cerebrospinal fluid cultures at 24 hours were sterile in all patients . Cefotaxime was safe and effective in treating Gram-negative enteric bacillary meningitis in infants and children and should be considered as a potential drug of choice in Gram-negative neonatal meningitis due to susceptible organisms.

Antonie Van Leeuwenhoek, 1988, 54(2), 95 - 111
Instability of protease production in a rel+/rel- -pair of Bacillus licheniformis and associated morphological and physiological characteristics; Bulthuis BA et al.; A naturally occurring relaxed/protease-producing (A-type) versus stringent/not protease-producing (B-type) pair of an industrial Bacillus licheniformis has been characterized; either of the two types can convert into the other . Both types can sporulate, grow anaerobically, grow at 56 degrees C and reduce nitrate; morphologically, they can easily be distinguished by cell- and colony-shape . They differ in the ability to use 12 substrates, as determined in API-tests . The two types are remarkably different in their content of extrachromosomal elements (A-type: 2; B-type: 4); furthermore, they differ in their rel-status (A-type: relaxed; B-type: stringent) . We propose that the differences in the ability of the two types to use different substrates probably are due to integration/extrusion of the extrachromosomal elements in and out of the chromosome, distorting or restoring a number of genes, together with induction of certain catabolic genes that are under control of the rel-system.

Biull Eksp Biol Med, 1988 Jan, 105(1), 27 - 30
{Ammonia release and binding processes and the activity of acid proteinase and glycolysis enzymes in the dynamics of experimental toxic adrenal encephalopathy}; Kovalenko VM et al.; The content of ammonium, glutamine, glutamate, aspartate and GABA, glutamine synthetase activity, acid proteinase, hexonase, phosphohexoisomerase and dehydrogenase glucose-6-phosphate were studied in dog brain homogenates after individual injections of Bacillus coli endotoxin (10 micrograms/kg) and adrenaline (75 micrograms/kg) into veins and their combined injections into the carotid artery . Isolated injections of endotoxin and adrenaline were shown to cause transient metabolic compensatory changes . Combined injections caused stable progressing brain metabolic disorders . It is suggested that neurochemical changes influence endogenous development of toxic adrenal encephalopathy.

Acta Stomatol Croat, 1988, 22(3), 169 - 75
{Scanning electron microscope study of early bacterial penetration of human enamel in initial caries}; Stanicic T et al.; Bacterial penetration of enamel during initial manifestations of the carious process was studied in intact buccal enamel of 8 impacted third permanent molars . After cleaning them from organic plaque, each buccal plane was cut into five segments, one of them serving as a control specimen and the other four being fixed into slots on partial prostheses of our volunteers . The specimens were left in oral cavity for 7, 14, 21 and 28 days, where they were exposed to the action of cariogenic factors . After removal of the organic plaque, the specimens were broken in two and the bacterial penetration into enamel was observed on the fractured cross-section using scanning electron microscope . In 4 specimens from the group orally exposed during a 7-day period, individual coccoid or bacilliform bacteria were found to have penetrated 5-10 microns deep . The number of bacteria and the depth of their penetration into enamel increased with the duration of oral exposition . Among the specimens orally exposed during 28 days, bacteria were observed to be present both individually and in colonies, penetrating to the depth of 60-90 microns . These bacteria could quite easily communicate with saliva and plaque via the pore system, which allowed them to produce metabolites, including lactic acids . This, in turn, allowed them to penetrate through the enamel, thus altering both the course and rate of the carious process progression.

Toxicon, 1988, 26(12), 1177 - 85
Kinetics of hemolysis induced by a toxin from Bacillus thuringiensis israelensis; Weinstein SA et al.; The kinetics of hemolysis resulting from the action on rabbit erythrocytes of a highly purified cytolytic toxin (26,000 mol . wt) isolated from a spore-crystal mixture of Bacillus thuringiensis israelensis was studied . Course of hemolysis, as determined by release of hemoglobin, yielded sigmoid curves whose maximum slopes were taken as a measure of the rate of lysis . Hemolysis occurred without an induction period, and the rate of lysis was a linear function of toxin concentration . Rate of hemolysis as a function of temperature yielded an Arrhenius constant of 9300 calories per mole . The toxin was active between pH 4.5 and 8.0 . Lysis was strongly inhibited by Cu2+, Fe2+ and Zn2+ in concentrations as low as 0.025 M . Phosphatidylserine, phosphatidylinositol, phosphatidylcholine and sphingomyelin inhibited lysis, whereas phosphatidylethanolamine, cerebroside, cholesterol and major integral erythrocyte membrane proteins caused little or no inhibition . Inhibition of lysis by sucrose indicates that hemolysis is of the colloid-osmotic type.

Eur Urol, 1988, 15(1-2), 146 - 9
Spontaneous regression of pulmonary metastases and long-term survival of a patient with metastatic renal cell carcinoma, after immunostimulation with bacillus Calmette-Guérin and extirpation of brain and contralateral lung metastases; Martinez-Pineiro L et al.; A 52-year-old patient underwent left radical nephrectomy for renal cell carcinoma and received adjuvant chemotherapy postoperatively with adriamycin and 5-fluoruracil . Twenty months afterwards he developed cerebral and multiple bilateral pulmonary metastases . The cerebral metastasis was excised and nonspecific immunostimulation with bacillus Calmette-Guerin was initiated . Two years later the other secondaries had scarcely grown and extirpation of the two right pulmonary metastases was undertaken . Few months afterwards the two left pulmonary nodules disappeared . The patient continues free of disease more than 5 years after nephrectomy and 16 months after regression of the lung metastases.

Dev Comp Immunol, 1988 Fall, 12(4), 707 - 17
Age related occurrence of natural agglutinins in the erisilkworm, Philosamia ricini; el Moataz Bellah M et al.; Natural agglutinins against human, guinea pig, and rat erythrocytes (RBC) and bacteria Bacillus thuringiensis were readily detected in hemolymph of final instar larvae with resulting titers of 0-9 (log2) . Titers were independent of sex and season but varied conspicuously and reproducibly with age . Moreover, response of hemagglutinins to heating varied with age being heat-labile in younger larvae, yet totally resistant to heating for 30 min at 70 degrees C in older ones . Bacterial agglutinins were uniformly resistant at all larval ages . The study thus reveals that the amounts and physico-chemical properties of P . ricini agglutinins change with larval development . Therefore, larval age should be taken into close consideration before reporting on the occurrence and properties of agglutinins in insects.

Ann Chir Main, 1988, 7(2), 166 - 75
{Various aspects of tuberculosis of the hand: apropos of a series of 45 cases}; Benkeddache Y et al.; Localisation of tuberculosis in the hand is a rare entity . We report here on 45 cases representing 7% of 650 cases of tuberculosis of the musculoskeletal system treated in our clinic during the past 16 years . Localisation in a joint or tendon sheath is classic but some others such as osteitis (8 cases) or soft tissue lesions (3 cases) have been seen, so the diagnosis must be proved bacteriologically by demonstrating the presence of the tubercle bacillus or, more often, histologically . A prolonged clinical course before the patient seeks medical advice is common and the clinical manifestations are numerous . Some patients were treated with a classical one year regimen . More recent cases have been treated with a short six month regiment . A comparison of the effectiveness of the two regimes is made.

Rev Chir Orthop Reparatrice Appar Mot, 1988, 74(3), 223 - 31
{Total hip arthroplasty in the presence of sepsis}; Vidal J et al.; Sixty-seven cases of total arthroplasty of the hip in the presence of sepsis performed between 1974 and 1986 were reviewed . These prostheses were inserted in 57 cases to replace septic prostheses . Forty-six were infected total hip prostheses and 11 were infected head and neck replacement femoral prostheses . In 10 cases arthroplasty was performed for subacute septic arthritis, usually secondary to trauma . In seven cases, a simple revision of the prosthesis by excision and lavage of infected tissues was performed . In 51 cases, a total hip replacement arthroplasty was performed in one stage . When one of the components was not loose it was preserved; in 17 cases the femoral cement was retained and in six the acetabular cup was retained . In eight cases, the prosthesis was removed, excision of the tissues was made and a total hip arthroplasty inserted after a minimum interval of six months . As regards infection, success was achieved in 69 per cent of cases rising to 85 per cent after secondary removal of the prosthesis . In relation to function, a good or fair result was achieved in 72 per cent of cases . The results were favourable in 81 per cent of cases with complete exchange of prostheses and in 75 per cent with preservation of the femoral cement . Preservation of the acetabular cup was only successful in 40 per cent of cases . Accessory factors in the prognosis were the addition of a bone graft, which became incorporated in 29 cases out of 30 and the nature of the organism which was of no significance, except that staphylococcus epidermidis and Koch's bacillus proved to be more easy to treat.

Cancer Invest, 1988, 6(3), 337 - 49
Active immunotherapy of human melanoma exploiting the immunopotentiating effects of cyclophosphamide; Berd D et al.; Malignant tumors may escape rejection by the immune system because they induce a state of immunological tolerance mediated by tumor antigen-specific suppressor T cells . In animal systems, cyclophosphamide can reverse the tolerance and thereby facilitate immunologically mediated tumor destruction . We have applied these concepts to the immunotherapy of human malignant melanoma . Forty-three patients with metastatic disease were treated with a whole cell vaccine 3 days after intravenous administration of cyclophosphamide, 300 mg/m2 . The vaccine consisted of cryopreserved, irradiated autologous melanoma cells, obtained from metastatic masses by dissociation with collagenase and DNAse, mixed with bacillus Calmette-Guerin (BCG) and injected intradermally . The cyclophosphamide (CY) + vaccine combination was repeated every 28 days . Delayed-type hypersensitivity (DTH) was tested by injecting 1 x 10(6) melanoma cells intradermally and measuring the diameter of induration at 48 h . Most patients had minimal pretreatment DTH responses to melanoma cells (mean +/- SE, mm = 2.4 +/- 0.5) . After two vaccine treatments, the responses increased significantly (mean increase +/- SE = 12.1 +/- 1.6 p less than .001) and that level of response was maintained after 4, 6, and 8 treatments . The patients were also skin-tested with a mixture of the enzymes used to dissociate the tumors . No patients exhibited DTH to collagenase + DNAse prior to vaccine injection, but every patient developed DTH to the mixture following two treatments (mean, mm = 26.4 +/- 3.9) . Although extracting viable cells from tumor tissue without the use of enzymes proved difficult, we were able to test DTH to mechanically dissociated tumor cells in 23 patients . After two vaccine treatments, there was a significant increase in DTH to enzyme-free autologous melanoma cells (mean DTH +/- SE, mm: 5.4 +/- 1.0, p less than .01) . Whereas 5 of 23 patients had positive DTH responses (5 mm induration or greater) before treatment, 11 of 23 were positive after two treatments . Further significant increases in DTH enzyme-free cells were observed after 6 and 8 treatments . Thus, it appears that patients receiving CY + vaccine developed DTH to tumor-associated antigens as well as to residual collagenase and DNAse on the cell surface . Thirty-three patients could be evaluated for antitumor effects of cyclophosphamide + vaccine . There were 3 complete remissions, 1 partial remission, and 2 minor responses . Two complete responders remain alive and free of disease after 57 and 12 months, respectively, and the third died after 39 months . The partial remission consisted of 75% regression of a pulmonary metastasis.(ABSTRACT TRUNCATED AT 400 WORDS)

Folia Microbiol (Praha), 1988, 33(6), 433 - 9
Inhibitory effect of 1-methyldodecyldimethylamine oxide and N,N-bis(dodecyldimethyl)-1,2-ethanediammonium dibromide on the spores of Bacillus cereus; Cupkova V et al.; 1-Methyldodecyldimethylamine oxide (MDDO) and N,N'-bis(dodecyldimethyl)-1,2-ethanediammonium dibromide (BDED) exhibit a significant affinity for the surface of Bacillus cereus spores and adsorb very rapidly to the cells; they have a pronounced inhibitory effect on spore outgrowth . In order to alter the affinity of the spore surface for these inhibitors, the spores were pretreated with sodium dodecyl sulfate (SDS), and with an electronegative (Tween 80) and electropositive (histone) compound . In SDS-pretreated spores the inhibitory effect of MDDO and BDED was abolished to a considerable extent . Whereas the development of intact spores was inhibited already after germination, in SDS-pretreated spores the postgermination development continued but was not completed . In Tween 80-pretreated spores the addition of BDED led only to a retardation of outgrowth and division; BDED added only during the division stage interrupted further development completely . Histone-pretreated spores stopped their development instantaneously after the addition of BDED at any phase of the postgermination development . The possible mechanisms of the interaction of the compounds used with spore surface or rather with the state of its structures are discussed.

Microbiol Immunol, 1988, 32(10), 991 - 8
Immunoelectron microscopic studies on spore coat proteins of Bacillus megaterium; Imagawa M et al.; An immunochemical staining technique for the spore coat proteins of Bacillus megaterium ATCC 12872 was developed using colloidal gold as a second antibody . For reducing the non-specific immunogold binding and increasing the specific binding, the affinity-purified IgG was used as a first antibody . In sporulating cells at t10, gold particles were found not only in the spore coat but also in the mother cell cytoplasm, suggesting that some coat proteins were synthesized in the cytoplasm . Use of the specific affinity-purified antibody to 48K-protein demonstrated that this protein was one of the components of the outer coat.

Biol Cell, 1988, 64(1), 3 - 11
Comparison between the molecular characteristics and the potential activity of X and Y nucleolar organizers from various Drosophila melanogaster laboratory lines; Rosenberg-Bourgin M et al.; The molecular characteristics of nucleolar organizers from X and Y chromosomes of different Drosophila melanogaster lines have previously been studied (17) . By analysis of appropriate genetic crosses we show in the present study that the X and Y chromosomes of these lines can confer different degrees of resistance on an inhibitor of ribosomal RNA synthesis (beta exotoxin or thuringiensin) present in the thermostable supernatant of Bacillus thuringiensis cultures . None of the lines studied gives rise to any particular phenotype under normal culture conditions; variations in the degree of supernatant resistance of these lines provide a relative measure of what can be called the potential activity of the nucleolar organizers of the different X and/or Y chromosomes . The potential activity of the Y nucleolar organizers is found to be generally higher than that of the X organizers . This result can be correlated with the fact that the number of uninterrupted ribosomal units is much greater on the Y chromosomes than on the Xs . Significant variations in potential activity have been shown to occur among the X as also among the Y nucleolar organizers . Comparison between the molecular characteristics of the nucleolar organizers and their level of activity shows that among the different ribosomal units, only those devoid of insertion interfere with the level of activity . However, some of our results could also indicate that not all the uninterrupted units have the same level of activity; this level could be related to the size of the nontranscribed spacer of the ribosomal units.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacotherapy, 1988, 8(6), 334 - 50
Aminoglycosides: current role in antimicrobial therapy; Cunha BA; Aminoglycosides remain the cornerstone of antibiotic therapy for nosocomial, gram-negative bacillary infections despite the recent introduction of broad-spectrum beta-lactam antibiotics and quinolones with antipseudomonal activity . Initially, aminoglycosides were used as antiaerobic gram-negative antimicrobial therapy . Currently, they have a key role in many types of infections, such as gram-negative urosepsis and in febrile granulocytopenic patients, because of their established antipseudomonal activity . Empiric treatment of febrile episodes in granulocytopenic cancer patients with an aminoglycoside, in combination with an anti-pseudomonal beta-lactam, accounts for much of the aminoglycoside use . Amikacin is emerging as one of the most effective aminoglycosides on the basis of resistance rates, pharmacokinetic factors likely to affect clinical efficacy, safety, and overall cost of therapy.

J Hyg Epidemiol Microbiol Immunol, 1988, 32(4), 419 - 23
Mycobacterial species responsible for tuberculosis and mycobacteriosis cases in the Czech Socialist Republic, 1981-1985; Kubin M et al.; This study is based on the analysis of nation-wide notification data on the bacteriologically proved mycobacterial infections registered in the national Information System on Bacillary Tuberculosis . Over the 5-year period under study, as many as 2,226 M . tuberculosis isolations were on average reported annually in the country (i.e . 21.6 per 100,000 population), and of these 62.5% were isolations in males, 63.5% were repeated isolations and 87% were isolations from sputum specimens; identification of human of M . bovis infection was reported in 13 patients annually only, mostly in those of higher age categories . The annual average of notified M . kansasii isolations was 138 (1.3 per 100,000 population), repeated isolations of the agent were reported in 66.6% of patients, mostly males 45-54 years old, and 81% of all reports came from an endemic area in the North-Moravian Region; notification of positive M . avium-intracellulare isolations occurred in 24 persons annually, in 48% of these repeatedly, M . xenopi isolation was reported in 87 and M . fortuitum/chelonei in 12 cases annually.

Crit Rev Biotechnol, 1988, 8(3), 217 - 24
Engineering thermostability in subtilisin BPN' by in vitro mutagenesis; Rollence ML et al.; A procedure has been developed for the isolation and identification of mutants of the bacterial serine protease, subtilisin, which exhibit enhanced thermostability . The cloned subtilisin BPN' gene from Bacillus amyloliquefaciens was treated with a variety of chemical mutagens to introduce random mutations in the coding sequence . Strains containing the cloned, mutagenized subtilisin gene which produced subtilisin with enhanced thermostability were selected by a simple plate assay procedure, which screens for esterase activity on nitrocellulose filters after preincubation at elevated temperatures . The identification and characterization of eight different stabilizing mutations are described . Several mutants containing various combinations of these stabilizing mutations were constructed by oligonucleotide-directed mutagenesis . Combining independent, stabilizing mutations in the same subtilisin molecule has resulted in an approximate multiplicative decrease in the rate of thermal inactivation . In this way, a variant of subtilisin has been constructed which is about 12-fold more stable than wild-type subtilisin, with no radical changes in the tertiary protein structure but rather minor, independent alterations in amino acid sequence . The ultimate goal in these studies is to be able to accurately predict where stabilizing changes can be made in a protein.

Microbiol Immunol, 1988, 32(9), 973 - 9
Isolation and characterization of outermost layer deficient mutant spores of Bacillus megaterium; Takubo Y et al.; Outermost layer deficient mutant spores of Bacillus megaterium ATCC 12872 were isolated by Urografin density gradient centrifugation after mutagenesis with ethyl methanesulfonate . Although the composition of the cortex peptidoglycan was the same as that of the parent spores, three major proteins (48, 36, and 22 K daltons) were missing, suggesting that these proteins are components of the outermost layer . All mutant spores were also found to have very hydrophobic surface by 'salt aggregation test,' which would facilitate selection of such mutants.

Zentralbl Mikrobiol, 1988, 143(5), 383 - 7
Protection of cadmium toxicity to B . cereus, E . coli and A . niger by Na+, Mg++ and Ca++ ions; Naidu CK et al.; 10ppm of cadmium apparently seem to extend the lag phase of Bacillus cereus and Escherichia coli but in reality both 10 and 100ppm decreased the growth with differing intensity . 1.0ppm of cadmium decreased the growth of Aspergillus niger, and it failed to grow at 500ppm . However, addition of calcium, magnesium and sodium gave increasing protection against cadmium toxicity for the growth of B . cereus, E . coli an A . niger . It is inferred that environment pollution by cadmium is related to the cationic availability in the environment.

Microbiol Immunol, 1988, 32(5), 529 - 33
Immunological detection of 22K protein in sporulating cells of Bacillus megaterium ATCC 12872; Nishihara T et al.; The synthesis and deposition of 22,000-dalton (22K) spore coat protein were examined immunochemically on the sporulating cells of Bacillus megaterium ATCC 12872 using the antibody to purified 22K spore coat protein . This antibody cross-reacted with 44K and 25K proteins in immunoblot analysis of dormant spore coat proteins . Immunoblot analysis on the sporulating cells showed that 22K protein was detected from t8 in forespore coat protein fractions . Sandwich enzyme immunoassay revealed that 22K protein in the spore coat protein fraction appeared at t6 and reached a plateau at t9, and 22K protein in the mother cell cytoplasmic fraction was detected at only t7 and t8 at a very low level.

Mol Cell Biochem, 1988 Jan, 79(1), 63 - 71
Characterization of the protein expressed in Escherichia coli by a recombinant plasmid containing the Bacillus megaterium cytochrome P-450BM-3 gene; Narhi LO et al.; In two previous reports (Narhi LO, Fulco AJ, J . Biol . Chem . 261: 7160-7169, 1986; Ibid., 262: 6683-6690, 1987) we described the characterization of a catalytically self-sufficient 119,000-dalton P-450 cytochrome that was induced by barbiturates in Bacillus megaterium . In the presence of NADPH and O2, this polypeptide (cytochrome P-450BM-3) catalyzed the hydroxylation of long-chain fatty acids without the aid of any other protein . The gene encoding this unique monooxygenase was cloned into Escherichia coli and the clone harboring the recombinant plasmid produced a protein that behaved electrophoretically and immunochemically like the B . megaterium enzyme (Wen LP, Fulco AJ, J . Biol . Chem . 262: 6676-6682, 1987) . We have now compared authentic P-450BM-3 from B . megaterium and putative P-450BM-3 isolated from transformed E . coli and have found them to be indistinguishable with respect to chromatographic and electrophoretic behavior, reaction with specific antibody, prosthetic group (heme, FAD and FMN) analyses, spectra, enzymology, limited trypsin proteolysis and partial amino acid sequencing . We thus conclude that the P-450 cytochrome expressed by the transformed E . coli is essentially identical to native P-450BM-3 induced by barbiturates in B . megaterium . The evidence furthermore suggests that the primary amino acid sequence of this complex protein is alone sufficient to direct the proper integration of the three prosthetic groups and to specify folding of the polypeptide into the correct tertiary structure.

Retina, 1988, 8(3), 182 - 4
Bacillus Calmette-Guerin (BCG) endophthalmitis; Lester H et al.; A patient who had been treated for metastatic bladder carcinoma with bacillus Calmette-Guerin (BCG) vaccine subsequently developed bilateral visual loss secondary to an infiltrative retinitis and vitreitis . Although assumed to have candida endophthalmitis, vitrectomy demonstrated mycobacterium bovis . The patient was placed on systemic anti-tuberculous therapy, and there was noted to be a reduction in retinal lesions prior to his death . This is the first report in the literature documenting endogenous endophthalmitis after BCG usage.

Braz J Med Biol Res, 1988, 21(3), 523 - 5
Murine delayed type hypersensitivity is suppressed by Ascaris suum extract; Macedo MS et al.; We studied the suppressive effect of an Ascaris suum extract on delayed-type hypersensitivity (DTH) reactions to ovalbumin (OVA) and to Bacillus Calmette-Guerin (BCG) . The ability of mice to develop DTH reactions to both antigens was suppressed when an immunizing dose of the antigen was given subcutaneously together with the Ascaris extract . Partial or complete suppression of the response to OVA was obtained by the use of 400 or 1000 micrograms of Ascaris extract, respectively . The response to BCG, on the other hand, was totally suppressed by 400 micrograms of extract.

Urol Res, 1988, 16(5), 351 - 5
BCG induced killer cell activity; Koga S et al.; To investigate the mechanism of Bacillus de Calmette Guerin (BCG) bladder instillation therapy, the killer cell activity induced in peripheral blood mononuclear cells (PBMNCs) after BCG instillation was examined . Significant cytotoxic activity against natural killer (NK) cell resistant target tumor cells was detected after 3 days of instillation . To characterize this BCG induced cytotoxic activity further, human PBMNCs were cultured with BCG in vitro . From 24 h maximum cytotoxicity was obtained and continued for 3 days, then decreased slightly . Neither a DNA synthesis inhibitor Cytosine-arabinoside (Ara-C) nor a cytotoxic T cell (CTL) generation inhibitor Cyclosporine A inhibited this killer cell activation . Monoclonal antibody treatment revealed that both precursor and effector cells are Leu1-, 3a-, 7+, 11b+ . The recognition specificity from cold target competition experiments was selective . Taken together NK type precursor was activated with BCG into NK type effector which has wider spectrum of target cells than usual NK cell.

Trans R Soc Trop Med Hyg, 1988, 82(2), 321 - 3
Isolation, purification and quantification of phenolic glycolipid-1 from human leprosy skin tissues; Venkatesan K et al.; Phenolic glycolipid-I, a marker lipid of Mycobacterium leprae, was isolated from skin biopsies obtained from untreated lepromatous leprosy patients by silicic acid and florisil column chromatography and purified by thin layer chromatography . Tissues with varying bacillary loads were analysed for their phenolic glycolipid content . A good correlation was observed between the bacillary population of the tissues and the phenolic glycolipid content.

Mikrobiologiia, 1988 Jan-Feb, 57(1), 107 - 13
{Potential use of the monosaccharide composition of bacteria for their identification}; Pomazanov VV et al.; The monosaccharide composition of cell hydrolysates can be used as a criterion for the chemical differentiation of gram-positive bacteria . The monosaccharide composition of six bacterial species belonging to the genus Bacillus has been determined using gas chromatography, mass spectrometry and computers . The qualitative composition was similar, glucose, galactose, ribose and glucosamine being the main components in all of the species . Some Bacillus species differed in their minor components . Although the monosaccharide composition appeared to be homogeneous, bacteria can be identified in terms of their carbohydrate profile using computers . To this end, the monosaccharide composition of bacterial cells is represented as a two-dimensional data file including the qualitative composition of components and the quantity of each component.

Immunopharmacol Immunotoxicol, 1988, 10(4), 579 - 96
Effect of BCG upon functional and phenotypic immune markers in rats bearing the Dunning R3327 MAT-LyLu prostatic adenocarcinoma; Rubenstein M et al.; Rats bearing (or not bearing) the Dunning R3327 MAT-LyLu prostatic adenocarcinoma were treated with Bacillus Calmette-Guerin (BCG) and evaluated for immune competence using functional and phenotypic markers . Tumor presence significantly depressed total T and helper T cell representation along with the helper/suppressor T cell ratio . Functional immunity, measured by phytohemmagglutinin (PHA) induced blastogenesis, was also significantly depressed . When BCG was administered to non-tumor bearing animals, it had no effect upon T cell subset distributions but significantly reduced PHA induced blastogenesis . BCG similarly administered to tumor bearing animals did not alter the depressed helper/suppressor T cell ratio found in tumor bearing rats, but did significantly elevate PHA induced blastogenesis . However, these elevated levels of functional immunity in BCG treated tumor-bearing rats remained significantly below normal . These data demonstrate a poor correlation between functional and phenotypic assessments of immune capability.

C R Seances Soc Biol Fil, 1988, 182(2), 181 - 5
{Use of acetamide by Bacillus gordonae . II . Research on acetamidase and the taxonomic value of spontaneous mutation permitting the acquisition of this enzyme}; Pichinoty F; The wild strain Q1 had no acetamidase . The mutant Q1Ac synthesized an inducible acetamidase which was catabolite repressible by glucose . The mutation described is a character that has a high taxonomic value . It constitutes a new example of acquisitive evolution.

C R Seances Soc Biol Fil, 1988, 182(2), 177 - 80
{Use of acetamide by Bacillus gordonae . I . Isolation and characterization of mutants able to use this compound as a source of carbon and energy}; Pichinoty F; All known strains of Bacillus gordonae can acquire, by spontaneous mutation, the ability to grow at the expense of acetamide as a source of carbon, nitrogen and energy . The isolation and characters of these mutants are described . Their frequency is high (10(-3)-10(-2) per cell).

J Gen Microbiol, 1988 Jan, 134 ( Pt 1), 97 - 105
Molecular cloning and nucleotide sequence of the cyclomaltodextrin glucanotransferase gene from the alkalophilic Bacillus sp . strain no . 38-2; Kaneko T et al.; The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from the alkalophilic Bacillus sp . strain no . 38-2 was cloned in Escherichia coli using pBR322 . A plasmid, pCS8, was isolated from a transformant producing CGTase and the cloned CGTase gene was found to be in a 5.3 kb DNA fragment . The nucleotide sequence of a 2.5 kb segment encoding the CGTase was determined . This segment showed an open reading frame which would encode a polypeptide of 712 amino acids . The pCS8 CGTase had the same enzymic properties as those of the extracellular CGTase produced by the alkalophilic Bacillus sp . strain no . 38-2 . The nucleotide and amino acid sequences of this CGTase gene and gene product, respectively, have strong homology with those of the Bacillus macerans CGTase.

Cancer Immunol Immunother, 1988, 26(1), 43 - 7
The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives . III . Eradication of disseminated murine chronic leukemia cells by utilizing MDP hapten-reactive helper T-cell activity; Shima J et al.; A previous paper has demonstrated that enhanced tumor-specific immunity could be induced by priming mice with Bacillus Calmette Guerin (BCG) and subsequently immunizing them with syngeneic tumor cells modified with BCG-cross-reactive muramyl dipeptide (MDP) hapten . The present study establishes a tumor-specific immunotherapy protocol for a murine chronic leukemia based on the above T-T cell collaboration between antitumor effector T cells and anti-MDP hapten helper T cells induced by BCG priming . BALB/c mice which had been primed to BCG were injected intravenously (i.v.) with viable, syngeneic BCL1 leukemia cells . One week later, these mice were immunized intraperitoneally (i.p.) with unmodified or MDP hapten-modified, 10,000 R X-irradiated BCL1 cells, followed by 4 booster immunizations at 5-day intervals . The administration of unmodified BCL1 tumor cells into BCG-primed mice failed to prevent them from tumor death due to the persistent growth of preinjected BCL1 cells . In contrast, the immunization of BCG-primed, BCL1 leukemia-cell-bearing mice with MDP-modified BCL1 cells resulted in a high growth inhibition of leukemia cells and protection of these mice from death by leukemia . It was also revealed that potent tumor-specific, T-cell-mediated immunity was generated in mice which survived in this immunotherapy model . Thus, these results indicate that administration of MDP hapten-modified, syngeneic leukemia cells into leukemia-bearing mice which have been primed with BCG results in potent tumor-specific, T-cell-mediated immunity attributable to preventing the growth of disseminated leukemic cells.

Appl Environ Microbiol, 1988 Jan, 54(1), 277 - 9
Structural relatedness between mosquitocidal endotoxins of Bacillus thuringiensis subsp . israelensis; Garduno F et al.; A mosquitocidal toxin gene, cloned from Bacillus thuringiensis subsp . israelensis, was introduced into mutant crystal-negative B . thuringiensis subsp . israelensis cells . Partial toxicity to mosquitos was restored . The 58-kilodalton cloned gene product is a minor protein component of B . thuringiensis subsp . israelensis crystals and is structurally related to a major, 135-kilodalton crystal toxin.

Mol Biother, 1988, 1(1), 30 - 6
Active specific immunotherapy with an autologous tumor cell vaccine in patients with resected non-small cell lung cancer; Schulof RS et al.; Eighteen postoperative patients with non-small cell lung cancer were actively immunized with a vaccine that included autologous cryopreserved irradiated tumor cells admixed with bacillus Calmette-Guerin . Patients received three weekly intradermal immunizations beginning 1-3 months after surgery (15 patients) or after completion of postoperative radiotherapy (3 patients) . There was marked heterogeneity in the relative proportion of tumor cells versus host infiltrating cells within individual vaccines (range of percent tumor cells 7-75%) . Five patients exhibited positive delayed cutaneous skin test reactivity (DCR) to autologous irradiated tumor cells prior to immunization, whereas 8 of 13 converted from skin test negative to positive . There were no correlations between DCR reactivity and in vitro lymphoproliferative responses to autologous tumor cells or to clinical outcomes, i.e., freedom from relapse . Possible explanations for the heterogeneity of the lung cancer vaccine and approaches for improving its immunogenicity are discussed.

J Basic Microbiol, 1988, 28(1-2), 11 - 6
Netropsin increases formation of mRNA coding for a neutral metalloproteinase in Bacillus megaterium; Chaloupka J et al.; The anticancer drug netropsin increases the synthesis of an exocellular metalloproteinase during exponential growth as well as in the stationary phase of a sporulating strain of Bacillus megaterium . Its effect is due to a stimulation of the synthesis of the mRNA coding for the proteinase, determined as a residual synthesis of the enzyme in the presence of actinomycin D . The half-life of the proteinase mRNA (5-6 min at 35 degrees C) is not affected by netropsin . Netropsin relieves partially the repression of the proteinase mRNA caused by amino acids, whereas the repression brought about by an increased temperature is almost unaffected by the drug.

Mol Microbiol, 1988 Jan, 2(1), 153 - 7
Differential specificity of two insecticidal toxins from Bacillus thuringiensis var . aizawai; Knowles BH et al.; Bacillus thuringiensis var . aizawai HD-249 produces more than one protein of 130-135 kD in its insecticidal crystal delta-endotoxin . We describe an indirect method of assessing the relative contribution to toxicity of two of these protoxins using monospecific antibodies directed against their active proteolytic products . Our results show that one toxin is active against Spodoptera frugiperda but not Choristoneura fumiferana cells in vitro, while the other lyses C . fumiferana but not S . frugiperda cells . There is no indication of synergism between these toxins in vitro.

Vet Pathol, 1988 Jan, 25(1), 72 - 6
Cilia-associated respiratory (CAR) bacillus infection of obese mice; Griffith JW et al.; Cilia-associated respiratory (CAR) bacillus was identified in respiratory tract lesions of obese mice dying of chronic respiratory disease . Neither Mycoplasma pulmonis nor pathogenic bacteria were isolated from cultures of the lesions at necropsy, but there was serologic and histologic evidence of respiratory virus infection . Cranial-ventral areas of lung were firm and demarcated from unaffected lung at gross examination, and representative tissue sank in water . Microscopically, there was suppurative bronchopneumonia with extensive peribronchiole lymphocyte and plasma cell proliferation . The affected bronchiole epithelium was covered with a sheet of slightly basophilic, filamentous, gram negative bacteria . Bronchioles with lesser amounts of lymphocyte accumulations contained lesser amounts of filamentous bacteria . Bronchioles without filamentous bacteria lining the respiratory epithelium lacked peribronchiole lymphocyte accumulations . There was a high correlation between CAR bacillus-positive serology and the identification of diagnostic histologic lesions . CAR bacillus was readily stained using immunohistochemical methods, and the ultrastructural features were similar to that described in rat infections.

Arkh Patol, 1988, 50(10), 63 - 7
{The morphology of mesenteric lymph nodes in Whipple's disease}; Zolotarevskii VB et al.; A patient aged 45 suffering from Whipple's disease is described . The disease has been diagnosed on the basis of morphologic examination of a biopsy specimen of the small intestine mesenteric lymph node . The patient had suffered from the disease for 14 years; during the last 6 months he developed symptoms of the malabsorption syndrome with diarrhea and steatorrhea . The lymph nodes contained numerous macrophages with PAS-positive glycoprotein granules in the cytoplasm, small cavities filled with lipids; electron microscopy has revealed bacillus-like bodies in the macrophages and outside the cells . After the disease has been diagnosed, the patient has been administered a course of tetracycline therapy, that resulted in an improvement of his status.

Microbiol Immunol, 1988, 32(9), 877 - 85
Primary role of NADH formed by glucose dehydrogenase in ATP provision at the early stage of spore germination in Bacillus megaterium QM B1551; Sano K et al.; Metabolic events involved in energy metabolism were studied in order to evaluate the ATP-forming ability of Bacillus megaterium QM B1551 spores at the very early stage of germination . When heat-activated spores were germinated on glucose as a sole substrate, its oxidation into gluconate (catalyzed by glucose dehydrogenase, EC 1.1.1.47), the accompanying NADH formation, oxygen uptake, and RNA synthesis were initiated immediately after germination, even when anaerobic breakdown of 3-phosphoglycerate (an ATP source for spores) and the subsequent glucose metabolism via the phosphorylating pathway were impaired by potassium fluoride (KF) . In contrast, fructose metabolism and the accompanying metabolic events did not begin until a few minutes after triggering of germination, and those events were entirely abolished by KF, indicating that fructose metabolism is initiated exclusively via its phosphorylation by the ATP derived from endogenous 3-phosphoglycerate . Thus those results provided further evidence for our previous proposal (Otani et al (1987) Microbiol . Immunol . 31: 967-974; Sano et al (1988) Biochem . Biophys . Res . Commun . 151: 48-52) that the first molecules of ATP in germinating spores can be efficiently generated via aerobic oxidation of NADH, which is formed by glucose dehydrogenase . Fluorescence monitoring of NADH in germinating spores also supported this conclusion.

Acta Leprol, 1988 Jan-Mar, 6(1), 7 - 16
{Bacterial sampling using lymph node puncture-aspiration . A study of 50 patients with leprosy}; Latapi-Contreras F et al.; Fine needle aspiration of lymph nodes was performed in 50 leprosy patients and was compared with usual techniques of bacillary smears taken from nasal mucosa, ear lobule and cutaneous lesion we found that the former was more sensible (30%) regarding bacteriologic and morphologic (34%) indices; moreover this proceeding was more sensible (50%) en patients with type II leprosy reaction . After 6 month-multi-therapy schedule in 3 patients, the morphologic index decreases 2 points (SFG) and bacteriologic index 1 + (mean value) . It is concluded that fine needle aspiration of lymph nodes is a useful method, because of its simplicity and low traumatic effects and its sensibility to follow-up treatment and reactional phases.

Antonie Van Leeuwenhoek, 1988, 54(1), 37 - 45
Effect of growth temperature and media composition on the fatty acid composition of Bacillus stearothermophilus AN 002; Bezbaruah RL et al.; The influence of growth temperature, media composition and cell age on the chemical composition of Bacillus stearothermophilus strain AN 002 has been determined . The total cellular protein decreased and the free amino acid content increased with growth temperature, in both exponential and stationary growth phase . The protein and free amino acid contents of cells were higher in the stationary phase than in the exponential phase, irrespective of growth temperature and media composition . The RNA content was only reduced in cells grown at 55 degrees C . No significant variations were observed in the DNA and carbohydrate contents with respect to growth temperature and cell age . The total lipid and fatty acid compositions on the other hand varied as a function of growth temperature, cell age and media composition . Differences in the relative concentrations of even, odd and branched chain fatty acids were noticed . No variation was observed in the antiiso and unsaturated fatty acids with respect to growth temperature . The unique variations in the fatty acid composition and total lipids at the growth temperature of 50 degrees C and their variations in the stationary growth phase seem to be characteristic for B . stearothermophilus AN 002.

Zh Mikrobiol Epidemiol Immunobiol, 1988 Jan, (1), 51 - 5
{Method of determining proteolytic activity by using a conjugate of bovine serum albumin with peroxidase}; Zherdev AV et al.; The authors propose a method for determination of proteolytic activity, based on the hydrolysis of peroxidase-labeled molecules of bovine serum albumin immobilized on the surface of polystyrene microassay plates with the subsequent determination of peroxidase activity on the carrier or in the solution . The optimum conditions for the sorption of the labeled substrate have been established . The method permits the determination of bacillary alkaline protease at a concentration of 01 . microgram/ml within 45 minutes . The determination of four proteases has demonstrated that this method shows good correlation with the routine one (r = 0.98), but is more sensitive and less time- and labor-consuming.

Eur J Biochem, 1987 Dec 30, 170(1-2), 253 - 8
Tyrocidine-induced modulation of the DNA conformation in Bacillus brevis; Bohg A et al.; Using the {3H}trimethylpsoralen photobinding method {Sinden, R.R., Carlson, J.O . & Pettijohn, D.E . (1980) Cell 21, 773-783}, a decrease in unrestrained torsional tension of DNA was detected in Bacillus brevis cells when they had entered the sporulation phase . This decrease in superhelicity was found in cells which synthesized the peptide antibiotic tyrocidine and which were stimulated to sporulate . Fluctuations in superhelicity probably reflect a highly complicated picture of tension-relaxing and tension-inducing activities . Addition of tyrocidine to vegetative cells reduced by one-half the torsional tension from DNA, whereas ethidium bromide relaxes DNA completely . Cross-links between DNA and tyrocidine were introduced with ultraviolet light in vitro and in vivo indicating that the modulation of the DNA conformation in the cell may in fact be due to a DNA-tyrocidine interaction . In a growing B . brevis culture exogenous {3H}tyrocidine could only be photobound to DNA after the cells had entered the sporulation phase . Our results could mean that the peptide antibiotic tyrocidine is active in B . brevis on the DNA level as one regulatory factor controlling DNA functions.

Biochemistry, 1987 Dec 29, 26(26), 8524 - 8
Investigation of transition-state stabilization by residues histidine-45 and threonine-40 in the tyrosyl-tRNA synthetase; Leatherbarrow RJ et al.; We have analyzed various mutations involving residues Thr-40 and His-45 in the tyrosyl-tRNA synthetase of Bacillus stearothermophilus . The utilization of binding energy in catalysis of tyrosyl adenylate formation from tyrosine and ATP was determined from the free energy profiles for the mutant enzymes . Our results confirm that the side chains of Thr-40 and His-45 provide a binding site for the pyrophosphoryl portion of the transition state of this reaction and for pyrophosphate in the reverse reaction . Deletion of these side chains destabilizes the transition-state by 4.9 and 4.1 kcal mol-1, respectively, consistent with a charged hydrogen-bonding interaction . To examine the role of His-45 further, we constructed the potentially conservative mutations His----Gln-45 and His----Asn-45 . Both mutant enzymes are debilitated compared with the native enzyme . The His----Gln-45 enzyme is more active than enzyme in which the complete side chain is deleted (His----Ala-45), and so in this location glutamine is a semiconservative replacement . In contrast, the His----Asn-45 mutation is significantly worse than simple deletion of the side chain, indicating that asparagine at this position causes active destabilization of the transition state compared to His----Ala-45 . The amide-NH2 of glutamine may be considered stereochemically equivalent to the epsilon-NH of histidine whereas the amide-NH2 of asparagine is comparable to the delta-NH of histidine . The results suggest that the epsilon-NH rather than the delta-NH group of His-45 is involved in the transition-state stabilization.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochim Biophys Acta, 1987 Dec 17, 894(3), 484 - 98
Purification, properties, and cellular localization of Euglena ferredoxin-NADP reductase; Spano AJ et al.; Ferredoxin-NADP reductase from Euglena gracilis Klebs var . Bacillaris Cori purified to apparent homogeneity, yields a typical 36 kDa and an unusual 15 kDa polypeptide on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, exhibits a typical flavoprotein spectrum, contains FAD, and catalyzes NADPH-dependent iodonitrotetrazolium-violet diaphorase, NADPH-specific ferredoxin-dependent cytochrome-c-550 reductase and NADPH-NAD transhydrogenase activities . Rabbit antibody to the purified FNR blocks these activities specifically and also blocks the iodonitrotetrazolium-violet diaphorase activity of Euglena chloroplast completely . The low iodonitrotetrazolium-violet diaphorase activity in the plastidless mutant, W10BSmL, is mitochondrial and is not specifically blocked by the ferredoxin-NADP reductase antibody . Dark-grown non-dividing (resting) wild-type Euglena cells show a 4-fold increase in ferredoxin-NADP reductase activity during greening at 970 lx . Half of the low ferredoxin-NADP reductase activity in dark-grown cells is initially soluble, but by the end of chloroplast development nearly all of the enzyme is membrane-bound . The binding of ferredoxin-NADP reductase on exposure to light correlates with the extent of thylakoid membrane formation . Immunoblots of wild-type extracts during greening indicate that the 15 kDa polypeptide increases in the same manner as the extent of reductase binding to thylakoid membranes.

Biochem J, 1987 Dec 15, 248(3), 657 - 62
beta-lactamase I from Bacillus cereus . Structure and site-directed mutagenesis; Madgwick PJ et al.; The sequence of the gene for beta-lactamase I from Bacillus cereus 569/H has been redetermined . Oligonucleotide-directed mutagenesis has been carried out, and the effects of the changes on the ampicillin-resistance of Escherichia coli TG1 expressing the mutant genes have been studied . Lysine-73, close to the active-site serine-70 and a highly-conserved residue, has been converted into arginine . This change had a large effect on activity, but did not abolish it . An even larger effect was found in the mutant in which glutamate-166 had been converted into glutamine; this had little or no activity . On the other hand, the conversion of glutamate-168 into aspartate gave fully active enzyme . Glutamate-166 is an invariant residue, but glutamate-168 is not . Alanine-123 has been replaced by cysteine, to give active enzyme; this change forms part of the plan to introduce a disulphide bond into the enzyme.

J Biol Chem, 1987 Dec 15, 262(35), 16778 - 85
Kinetics and mechanism of inactivation of the RTEM-2 beta-lactamase by phenylpropynal . Identification of the characteristic chromophore; Grace ME et al.; beta-Lactamases of all three classes, A, B, and C, are inactivated by phenylpropynal and p-nitrophenylpropynal . The inactivation of RTEM-2 beta-lactamase and of Bacillus cereus beta-lactamase I is accelerated in the presence of A type substrates such as dicloxacillin, quinacillin, and cefoxitin, which are thought to expand or loosen the conformation of these enzymes . In the presence and absence of cefoxitin the inactivation of the RTEM-2 beta-lactamase is first and second order, respectively, in phenylpropynal concentration . The additional phenylpropynal molecule in the latter case may serve the same function as cefoxitin, viz . catalyze access to sensitive functional groups . Correlation of the loss of activity of the RTEM-2 enzyme with the extent of modification suggests that the modification of any one of about four kinetically equivalent groups leads to inactivation . Modification of all of the above mentioned enzymes leads to formation of a characteristic chromophore of unusual stability to nucleophiles, which absorbs maximally between 315 and 320 nm . A consideration of the properties of model compounds demonstrated that the protein-bound chromophore is that of a 1-phenyl-3-imino-1-propen-1-ammonium ion (Formula: see text), formed by reaction of phenylpropynal with two enzymic amine groups, and thus cross-linking the enzyme intramolecularly . Phenylpropynal may be a convenient general reagent for rapid and stable intramolecular cross-linking of proteins through lysine.

J S Afr Vet Assoc, 1987 Dec, 58(4), 183 - 6
The use of a diffusion test for the detection of antibiotics in the tissues of slaughter stock; Lloyd DN et al.; The Brilliant Black Reduction Test Kit (BR Test), which is widely used to detect antimicrobial residues in milk, was adapted to detect residues in the meat and tissues of slaughter stock . The adaptation consisted of placing kidney and muscle tissue samples into 2.5ml diffusion cups containing 0.4ml media plus Bacillus stearothermophilus spores and brilliant black indicator . A preliminary trial undertaken to test the lower limits of sensitivity of the adapted BR Test to a number of the more common antibiotics used in food animals, was followed by a survey involving 943 pigs slaughtered at one abattoir . Samples were tested from 87 suppliers of which 11 regularly marketed pigs with detectable antimicrobial residues . Most of these pigs came from large pig producing units . Three suppliers marketed pigs with suspicious reactions . No residues could be detected in pigs from the remaining 73 suppliers . The BR Test was found to be a quick, inexpensive, practical screening test which could be utilized for the routine detection of antimicrobial residues in slaughter stock at all South African abattoirs.

J Am Mosq Control Assoc, 1987 Dec, 3(4), 583 - 8
Floating bait formulations increase effectiveness of Bacillus thuringiensis var . israelensis against Anopheles larvae; Aly C et al.; The development and screening of floating-type bait formulations designed to improve the activity of bacterial toxins against larval Anopheles is described . Floating and spreading abilities of carrier particles (wheat flour) were compared using corn oil, lecithin, and two products yielding surface films on water (Arosurf and Liparol) . Mixtures containing 1 or 5% Arosurf showed the best spreading abilities on a water surface, but strongly inhibited the ingestion of wheat flour by Anopheles albimanus larvae . Corn oil and lecithin improved spreading satisfactorily at a concentration of 5% and inhibited larval feeding by only 6-25% . To select a suitable concentration of active ingredient in formulations, Bacillus thuringiensis var . israelensis (B.t.i.) primary powder in concentrations ranging from 0.01 to 0.5% was mixed with wheat flour/corn oil mixtures and tested in quantities exceeding the gut volumes of treated larvae . Complete mortality was obtained with concentrations of 0.1% (Anopheles stephensi), 0.2% (An . albimanus), or 0.3% (An . quadrimaculatus) B.t.i . When in 175-liter containers the activity of formulations (5% corn oil, 0.2% B.t.i.) and of toxin suspensions was compared by conventional dosage-mortality regression, formulations were more active by a factor of 68 against An . stephensi, 39 against An . albimanus and 67 against An . quadrimaculatus.

J Am Mosq Control Assoc, 1987 Dec, 3(4), 579 - 82
Efficacy of Bacillus sphaericus and Bacillus thuringiensis var . israelensis for control of Culex pipiens and floodwater Aedes larvae in Iowa; Berry WJ et al.; Granular and flowable concentrate formulations of Bacillus thuringiensis var . israelensis provided a 90-100% reduction in Aedes vexans and Culex spp . larvae in natural larval habitats . A briquet formulation of B . thuringiensis was less effective, providing a 12-76% reduction . No residual activity occurred in sites treated with B . thuringiensis . Granular formulations of Bacillus sphaericus (2.78-8.42 kg/ha) caused a 100% reduction in Culex pipiens larvae in natural sites and artificial pools . Bacillus sphaericus also controlled (84-98% reduction) a mixed population of Aedes trivittatus and Cx . pipiens in subplots of a retention pond . In field sites, B . sphaericus continued to control Cx . pipiens larvae for 96 hr.

Cell Mol Neurobiol, 1987 Dec, 7(4), 413 - 24
Vasopressin stimulates the phosphorylation of an 83,000 Mr protein in the superior cervical ganglion; Cahill AL et al.; 1 . 32P-Labeled proteins from the superior cervical ganglion of the rat were separated by two-dimensional gel electrophoresis and visualized by autoradiography . 2 . The most heavily labeled phosphoprotein in the ganglion had a relative molecular weight of 83,000 and a pI of 4.5 . Phosphorylation of this protein was increased by phorbol 12,13-dibutyrate, an activator of the Ca2+/phospholipid-dependent protein kinase, protein kinase C . This protein appears to be similar or identical to a specific protein kinase C substrate that has been described in other tissues (Blackshear, P . J., et al., J . Biol . Chem . 261:1459-1469, 1986) . 3 . Phosphorylation of this protein was also increased by treatment of the ganglion with phospholipase C (Bacillus cereus) but was not increased by 8-bromo-cyclic AMP or by nicotinic agonists . Vasopressin increased the hydrolysis of inositol-containing phospholipids in the ganglion and also increased the labeling of the 83,000 Mr protein . Thus, vasopressin appears to activate protein kinase C in the ganglion . 4 . Muscarine, which also increased phospholipid metabolism in the ganglion, did not increase the phosphorylation of the 83,000 Mr protein . Muscarine and vasopressin stimulate phospholipid metabolism in different structures within the ganglion (Horwitz, J., et al., J . Pharmacol . Exp . Ther . 237:312-317, 1986) . Muscarine may increase phospholipid metabolism in structures that do not contain significant amounts of the 83,000 Mr protein.

Biol Reprod, 1987 Dec, 37(5), 1170 - 8
Sperm-macrophage interaction in the mouse: a quantitative assay in vitro using 111indium oxine-labeled sperm; Olive DL et al.; The role of reproductive tract macrophages in contraception and reproductive failure has become widely recognized . However, in vitro analysis of sperm phagocytosis by macrophages has relied upon a semi-quantitative method of sperm counting that is of limited accuracy and reproducibility . We have developed an assay using murine sperm labeled with 111indium oxine, and results indicate the labeling to be rapid and efficient . Incorporation of 111indium into sperm increased the dose and sperm concentration and reached 90% maximal uptake after 15 min incubation, with maximal uptake occurring at 30 min . No decrease in sperm motility was noted with levels of oxine in excess of those required for significant labeling . Maximal labeling efficiency occurred in phosphate-buffered saline (PBS), with Dulbecco's modified Eagle's medium (DMEM) + 10% adult bovine serum (ABS) producing significantly less uptake . Label dissociation was detectable in PBS at room temperature, but at 37 degrees C in DMEM + 10% ABS, loss of label occurred at a rate of 23.5%/h . Addition of labeled sperm to murine macrophage monolayers under optimal conditions resulted in uptake of 111indium by macrophages, while free label was unincorporated . Results indicated assay specificity for macrophage-limited uptake, with insignificant label uptake by nonphagocytic murine fibroblasts and better sensitivity than sperm counting . Macrophages from Bacillus Calmette-Guerin (BCG)-infected mice resulted in a decrease in sperm uptake . Female macrophages showed greater capacity for sperm uptake than those of the male mouse . These initial studies demonstrated the utility of this model system in enhancing the understanding of sperm-macrophage interaction in the female reproductive tract.

J Environ Sci Health B, 1987 Dec, 22(6), 691 - 720
Influence of surfactants and polymeric adjuvants on physicochemical properties, droplet size spectra and deposition of fenitrothion and Bacillus thuringiensis formulations under laboratory conditions; Sundaram A et al.; The effect of two surfactants and two polymeric adjuvants on droplet size spectra and deposition patterns of nine spray formulations was investigated following atomization in a laboratory chamber using a spinning disc atomizer that can produce a narrow droplet size spectrum . Spray droplets were sampled using Kromekote cards and deposit recoveries were examined on glass plates . Physicochemical properties studied were: viscosity-shear rate relationship, surface tension, volatility, pH, conductance, electrophoretic mobility and zeta potential . Formulations containing low surfactant concentrations provided Newtonian liquids with low viscosities . These liquids atomized into small droplets and provided low recoveries of spray deposits on sampling units . However, formulations containing polymeric adjuvants, and a high concentration of a non-ionic surfactant provided pseudoplastic liquids with high viscosities . These formulations resulted in large droplets with high recoveries of spray deposits on sampling units . Among the physicochemical properties studied, viscosity, surface tension, volatility and electrophoretic mobility, played important roles on liquid atomization and droplet deposition.

J Biol Response Mod, 1987 Dec, 6(6), 610 - 24
Active specific immunotherapy of renal cell carcinoma patients: a prospective randomized study of hormono-immuno-versus hormonotherapy . Preliminary report of immunological and clinical aspects; Adler A et al.; Early results of a prospective, randomized trial of active, specific immunotherapy adjunctive to nephrectomy in all stages of RCC are presented . Forty-three patients with median followup of 30 m, who were randomly allocated to either immuno-hormonotherapy arm (IMT), or hormonotherapy alone (HT), are evaluated in terms of progression-free interval (PFI) and overall survival by life table method . Immunotherapy consisted of autologous irradiated tumor cells (AITC), admixed with bacillus Calmette-Guerin (Glaxo) administered by the intradermal and endolymphatic route . Clinical results of this study show only a trend for advantage of the experimental (IMT) arm over the control (HT) arm, this trend did not reach statistical significance level: prolongation of disease free period in stages I-III with localized disease (p less than 0.1) and prolongation of survival in patients with metastatic disease (p less than 0.07) . A correlation was established between induction of cutaneous delayed hypersensitivity (DTH) to AITC and prolonged PFI and survival: patients with positive DTH had a significantly better course of disease than those who could not be converted to positivity after repeated immunizations . Positive in vitro leukocyte migration inhibition against autologous tumor preparations correlates well with positive in vivo cutaneous DTH . Some immunological aspects of active immunization with autologous tumor cells are discussed.

Aust N Z J Med, 1987 Dec, 17(6), 568 - 73
The serological response to the phenolic glycolipid of Mycobacterium leprae in Australian and Nepali leprosy patients; Britton WJ et al.; Antibodies to the species-specific phenolic glycolipid (PGL-1) of Mycobacterium leprae and a crude M . leprae sonicate were measured by ELISA in sera from newly diagnosed and treated leprosy patients from Sydney and Nepal . IgM anti-PGL-1 antibodies were present in 88-90% of untreated patients at the lepromatous pole of the clinical spectrum and 35-55% of those at the tuberculoid pole . In treated patients with either form of the disease, IgM anti-PGL-1 antibodies were within the normal range or minimally elevated . In contrast, high levels of IgG anti-PGL-1 antibodies were detected in both treated and untreated patients . Neither IgM nor IgG anti-PGL-1 antibodies were elevated in sera from Mantoux negative controls and only one out of 15 sera from patients with untreated tuberculosis contained significant amounts of antibody . Comparison of the data from the anti-PGL-1 assay with the antibody response to a crude M . leprae sonicate revealed that the latter assay yielded more variable results and discriminated less well between lepromatous and tuberculoid subjects and between untreated patients and those on therapy . Thus the IgM anti-PGL-1 response signifies the presence of active disease, particularly in multi-bacillary cases, and has the potential to be used not only to monitor the response of these patients to therapy, but also to detect subclinical leprosy in high-risk groups such as the relatives of patients with lepromatous disease.

Urology, 1987 Dec, 30(6), 520 - 8
Superficial bladder cancer treated by intravesical bacillus Calmette-Guerin or adriamycin: multicenter study interim report; Khanna OP et al.; One hundred sixteen patients with superficial bladder cancers (Stages Ta, T1, and TIS) were evaluated and treated with either intravesical bacillus Calmette-Guerin {Tice strain} (BCG) or doxorubicin hydrochloride (Adriamycin {ADR}), in a multicenter study . One hundred nine of these patients currently have follow-up . Of these, 54 were completely resected and 55 incompletely resected . For complete resections, based on recurrence rates per 100 patient months, both BCG (0.22) and ADR (0.91) worked well, although BCG had a slightly lower recurrence rate . However, for incomplete resections, BCG (0.20) had a markedly lower recurrence rate than ADR (2.52) . Eighteen patients failed initial treatment, with either BCG or ADR . All have been placed on long-term therapy schedules . Of the 12 failures who currently have follow-up, 11 (92%) have either partially or completely responded with additional intravesical therapy . No patients in this study have yet required cystectomies.

Arch Biochem Biophys, 1987 Dec, 259(2), 466 - 72
Phosphorylase-mediated mobilization of the amino group of adenine in Bacillus cereus; Mura U et al.; Mobilization of the ribose moiety of purine nucleosides as well as of the amino group of adenine may be realized in Bacillus cereus by the concerted action of three enzymes: adenosine phosphorylase, adenosine deaminase, and purine nucleoside phosphorylase . In this pathway, ribose-1-phosphate and inorganic phosphate act catalytically, being continuously regenerated by purine nucleoside phosphorylase and adenosine phosphorylase, respectively . As a result of such a metabolic pathway, adenine is quantitatively converted into hypoxanthine, thus overcoming the lack of adenase in B . cereus.

Scand J Immunol, 1987 Dec, 26(6), 683 - 90
In vivo activated lymphoid cells (IVALC) affect the cloning efficiency of human T lymphocytes reactive to a soluble antigen, purified protein derivative; Sia DY et al.; Peripheral blood mononuclear cells (PBMC) of normal individuals were found to contain a proportion (4-9%) of in vivo activated lymphoid cells (IVALC) . These IVALC were characterized by their expression of interleukin 2 (IL-2) receptors, and by the ability to proliferate in the presence of exogenous IL-2 . There was a good correlation between the proportion of IVALC in different cell populations and the level of cell proliferation to IL-2 . It was found that IVALC isolated from autologous PBMC of Bacillus Calmette-Guerin (BCG)-immunized individuals contained no significant proportion of purified protein derivative (PPD)-reactive lymphocytes . The addition of IVALC markedly enhanced proliferative responses of the autologous T4+T8-IL-2 receptor-negative cell cultures to antigen stimulation . An increased proportion of activated (IL-2 receptor-positive) lymphocytes was generated in PBMC as compared to autologous T4+T8-IL-2 receptor negative cell cultures after stimulation with PPD . Limiting dilution analysis showed that IL-2 responsive IVALC through expansion markedly affected the cloning efficiency of antigen-proliferating T cells of autologous PPD-stimulated PBMC cultures . Only 1 out of every 11-25 blast cells generated in the PBMC cultures could establish itself as a growing colony based on determinations in six BCG-positive individuals . By using a T4+T8- population depleted of IVALC to generate PPD-reactive lymphocytes, a three- to four-fold increase in the cloning efficiency of antigen-specific cells was obtained.

Mol Gen Mikrobiol Virusol, 1987 Dec, (12), 16 - 20
{Isolation and characteristics of a plasmid from the thermotolerant strain of Bacillus licheniformis}; Gaidenko TA et al.; A 8.3 kb cryptic plasmid was isolated from the thermotolerant strain of Bacillus licheniformis 28KA and designated pLT83 . The replicative (rep) region was localized on the plasmid map . The pLT83 plasmid labelled in vitro with an antibiotic resistance determinant is able to replicate in B . subtilis cells . The pLT83 plasmid replicates stably in B . licheniformis strain at higher temperatures (37-60 degrees C) than in B . subtilis cells (37-50 degrees C) . The plasmid and its derivatives may be used as vectors for gene cloning in B . subtilis and B . licheniformis cells.

Appl Environ Microbiol, 1987 Dec, 53(12), 2808 - 14
Use of oligonucleotide probes to study the relatedness of delta-endotoxin genes among Bacillus thuringiensis subspecies and strains; Prefontaine G et al.; Fifteen Bacillus thuringiensis strains representing 13 serotypes were screened with five oligodeoxyribonucleotide probes specific for certain regions of two published sequences and one unpublished sequence of B . thuringiensis delta-endotoxin genes . Of the 15 cultures, 14 hybridized with at least one probe; the B . thuringiensis subsp . thompsoni strain alone did not hybridize . Two B . thuringiensis subsp . kurstaki strains of commercial interest, HD-1 and NRD-12, were found to be so closely related as to be indistinguishable with this technique; the same situation was found with strains from B . thuringiensis subspp . dendrolimus and sotto . Five strains were identified as probably containing only one endotoxin gene . A probe specific for the gene from the B . thuringiensis subsp . kurstaki HD-73 strain hybridized to only 3 of the 15 cultures tested . The hybridization data suggest that the DNA sequences coding for the C-terminal region of the endotoxin protein are as well conserved as those coding for the N-terminal toxic portion.

Am J Pathol, 1987 Dec, 129(3), 567 - 77
Acquisition of peroxidase activity by rat alveolar macrophages during pulmonary inflammation; Shellito J et al.; The authors investigated the ability of rat alveolar macrophages to acquire peroxidase activity in the course of pulmonary inflammation . Granulomatous pulmonary inflammation was induced in bacille Calmette-Guerin (BCG)-immunized rats by intravenous injection of BCG in mineral oil . In contrast to normal alveolar macrophages, which are peroxidase-negative, alveolar macrophages lavaged from the BCG-treated rats showed significant peroxidase activity in large cytoplasmic inclusions compatible with internalized exogenous material . Alveolar macrophage uptake of intact peroxidase-positive neutrophils was also observed . Maximal numbers of peroxidase-positive alveolar macrophages were observed after the initial influx of neutrophils into the lungs, and peroxidase activity could be demonstrated in cell-free lavage fluid during the acute phase of lung injury . Normal alveolar macrophages acquired peroxidase activity after incubation with peritoneal exudate neutrophils, with purified soluble human myeloperoxidase, and with opsonized erythrocytes . It is concluded that alveolar macrophages acquire peroxidase activity from multiple sources during pulmonary inflammation . Internalization of peroxidase by the alveolar macrophage may serve to clear a potentially toxic enzyme(s) from the alveolar space and contribute to the resolution of pulmonary inflammation.

J Bacteriol, 1987 Dec, 169(12), 5761 - 5
Bioenergetic properties of alkalophilic Bacillus sp . strain C-59 on an alkaline medium containing K2CO3; Kitada M et al.; Alkalophilic Bacillus sp . strain C-59 could grow well on an alkaline medium containing K2CO3, as well as Na2CO3, but did not grow on K+-depleted medium . Right-side-out membrane vesicles, energized in the absence of Na+, however, could not take up {14C}methylamine actively, while vesicles equilibrated with 10 mM NaCl actively took up {14C}methylamine . The uptake of {14C}serine was also stimulated by the addition of Na+, and the imposition of a sodium gradient caused transient uptake . These results indicated that an Na+/H+ antiporter was involved in pH homeostasis and generation of an electrochemical sodium gradient in strain C-59 even though a growth requirement for Na+ was not evident . The efflux of 22Na+ from 22Na+-loaded vesicles was more rapid at pH 9.5 than at pH 7 in the presence of an electron donor . On the other hand, vesicles at pH 7 showed more rapid efflux than at pH 9.5 when the antiporter was energized by a valinomycin-mediated K+ diffusion potential (inside negative).

Biochemistry, 1987 Nov 17, 26(23), 7409 - 18
Enzymatic hydrolysis of short-chain lecithin/long-chain phospholipid unilamellar vesicles: sensitivity of phospholipases to matrix phase state; Gabriel NE et al.; Short-chain lecithin/long-chain phospholipid unilamellar vesicles (SLUVs), unlike pure long-chain lecithin vesicles, are excellent substrates for water-soluble phospholipases . Hemolysis assays show that greater than 99.5% of the short-chain lecithin is partitioned in the bilayer . In these binary component vesicles, the short-chain species is the preferred substrate, while the long-chain phospholipid can be treated as an inhibitor (phospholipase C) or poor substrate (phospholipase A2) . For phospholipase C Bacillus cereus, apparent Km and Vmax values show that bilayer-solubilized diheptanoylphosphatidylcholine (diheptanoyl-PC) is nearly as good a substrate as pure micellar diheptanoyl-PC, although the extent of short-chain lecithin hydrolysis depends on the phase state of the long-chain lipid . For phospholipase A2 Naja naja naja, both Km and Vmax values show a greater range: in a gel-state matrix, diheptanoyl-PC is hydrolyzed with micellelike kinetic parameters; in a liquid-crystalline matrix, the short-chain lecithin becomes comparable to the long-chain component . Both enzymes also show an anomalous increase in specific activity toward diheptanoyl-PC around the phase transition temperature of the long-chain phospholipid . Since the short-chain lecithin does not exhibit a phase transition, this must reflect fluctuations in head-group area or vertical motions of the short-chain lecithin caused by surrounding long-chain lecithin molecules . These results are discussed in terms of a specific model for SLUV hydrolysis and a general explanation for the "interfacial activation" observed with water-soluble phospholipases.

J Biol Chem, 1987 Nov 15, 262(32), 15323 - 6
X-ray crystallographic studies of the alanine-specific racemase from Bacillus stearothermophilus . Overproduction, crystallization, and preliminary characterization; Neidhart DJ et al.; To facilitate large-scale purification and crystallographic study, we have subcloned the gene for the alanine racemase of Bacillus stearothermophilus from pICR401 (Inagaki, K., Tanizawa, K., Badet, B., Walsh, C . T., Tanaka, H., and Soda, K . (1986) Biochemistry 25, 3268-3274) and overproduced the enzyme in Escherichia coli W3110 lacIq using the tac promoter of PKK223-3 . This system yields alanine racemase as 6% of the bacterial cytosolic protein . Purification by a modification of the procedure of Inagake et al . yielded 75 mg of homogeneous alanine racemase from 30 g of cells (wet weight) . Large, well-formed crystals of alanine racemase have been grown from polyethylene glycol 8000 using vapor diffusion . These crystals have unit cell dimensions a = 85.3 A, b = 110.0 A, and c = 89.9 A . The crystals belong to space group P2(1), with beta fortuitously equal to 90 degrees within experimental error; however, they are frequently twinned by second order pseudomerohedry with twin fraction (the ratio of the volume of the smaller twin domain to the total volume of the crystal) ranging from about 0 to 0.5 . Fortunately, for crystals with low twin fraction, computational methods have been developed for the analysis and correction of simple twinning (Fisher, R . G., and Sweet, R . M . (1980) Acta Crystallogr . A36, 755-760) . The crystals contain two alpha 2 dimers of alanine racemase in the asymmetric unit . We have identified several potentially useful heavy atom derivatives in low resolution screening experiments and are proceeding with high resolution data collection.

Anal Biochem, 1987 Nov 15, 167(1), 113 - 7
The growth of ordered two-dimensional sheets of ribosomal particles from salt-alcohol mixtures; Arad T et al.; A procedure for the in vitro growth of well-ordered two-dimensional sheets from ribosomal particles using salts and salt-alcohol mixtures has been developed . Employing this procedure, ordered two-dimensional sheets of the wild type as well as of mutated 50 S ribosomal subunits from Bacillus stearothermophilus can readily be obtained . These sheets, stained with uranyl acetate or gold-thioglucose, are suitable for three-dimensional image reconstruction . They consist of relatively small unit cells with dimensions of 160 +/- 15 and 365 +/- 20 A . Diffraction patterns of electron micrographs of these sheets contain features to 25 A resolution.

Biochem J, 1987 Nov 15, 248(1), 181 - 8
An X-ray-crystallographic study of beta-lactamase II from Bacillus cereus at 0.35 nm resolution; Sutton BJ et al.; Crystals of beta-lactamase II (EC 3.5.2.6., 'penicillinase') from Bacillus cereus were grown with Cd(II) in place of the natural Zn(II) cofactor and stabilized by cross-linking with glutaraldehyde . Their space group is C2, the cell dimensions are a = 5.44 nm, b = 6.38 nm, c = 7.09 nm and beta = 93.6 degrees, and there is one molecule in the asymmetric unit . Diffraction data were collected from cross-linked crystals of the Cd(II)-enzyme, the apoenzyme and six heavy-atom derivatives . The electron-density map calculated at 0.35 nm resolution reveals the essential Cd(II) ion surrounded by three histidine residues and one cysteine residue . The position of a glutamic acid residue, modification of which destroys activity {Little, Emanuel, Gagnon & Waley (1986) Biochem . J . 233, 465-469}, suggests the probable location of the active site of the enzyme . Two minor Cd(II) sites not essential for activity were also located . The structure of the apoenzyme at this resolution appears to differ from that of the Cd(II)-enzyme only in the orientation of two of the histidine residues and the cysteine residue that surround the metal ion.

Biochem J, 1987 Nov 15, 248(1), 197 - 201
Analysis of the molecular basis of insecticidal specificity of Bacillus thuringiensis crystal delta-endotoxin; Haider MZ et al.; The mechanism of action and receptor binding of a dual-specificity Bacillus thuringiensis var . aizawai ICl delta-endotoxin was studied using insect cell culture . The native protoxin was labelled with 125I, proteolytically activated and the affinity of the resulting preparations for insect cell-membrane proteins was studied by blotting . The active preparations obtained by various treatments had characteristic specificity associated with unique polypeptides, and showed affinity for different membrane proteins . The lepidopteran-specific preparation (trypsin-treated protoxin containing 58 and 55 kDa polypeptides) bound to two membrane proteins in the lepidopteran cells but none in the dipteran cells . The dipteran-specific preparation (protoxin treated sequentially with trypsin and Aedes aegypti gut proteases, containing a 53 kDa polypeptide) bound to a 90 kDa membrane protein in the dipteran (A . aegypti) cells but bound to none in the lepidopteran cells or Drosophila melanogaster cells . The toxicity of trypsin-activated delta-endotoxin was completely inhibited by preincubation with D-glucose, suggesting a role for this carbohydrate in toxin-receptor interaction . The toxicity was also decreased by osmotic protectants to an extent proportional to their viscometric radius . These results support a proposal that initial interaction of toxin with a unique receptor determines the specificity of the toxin, following which cell death occurs by a mechanism of colloid osmotic lysis.

Biochim Biophys Acta, 1987 Nov 13, 904(2), 301 - 8
Na+ modulates the K+ permeability and the membrane potential of alkalophilic Bacillus; Matsukura H et al.; In the absence of Na+ in the medium, the membrane potential of obligately alkalophilic Bacillus cells was found to be decreased by the addition of K+ to the medium, whereas K+ addition in the presence of Na+ had no effect . Rb+ showed essentially the same effect as K+ . The decreased membrane potential was quickly restored by lowering the K+ concentration in the medium or by adding Na+ or Li+ to the medium . Thus, in the absence of Na+, the membrane potential of alkalophilic Bacillus seems to be affected by the concentration difference of K+ between inside and outside of the cell, and Na+ or Li+ in the medium suppresses the K+ effect . An exchange between extracellular Rb+ and intracellular K+ was observed in the absence of Na+ . However, the exchange was greatly suppressed by the addition of Na+ or Li+ to the medium, indicating that Na+ in the medium modulates the K+ permeability of the alkalophilic Bacillus cell membrane . The K+-induced decrease in the membrane potential of alkalophilic Bacillus in the absence of Na+ is accounted for by the increased K+-permeability of the cell membrane.

Biochim Biophys Acta, 1987 Nov 5, 916(1), 145 - 8
The engineering of a more thermally stable lactate dehydrogenase by reduction of the area of a water-accessible hydrophobic surface; Wigley DB et al.; A site-directed mutant of Bacillus stearothermophilus lactate dehydrogenase (lactate:NAD+ oxidoreductase, EC 1.1.1.27) has been engineered in which the conserved hydrophobic residue isoleucine-250 has been replaced by the more hydrophilic residue asparagine . This isoleucine forms a large part of a water-accessible, hydrophobic surface in the active site of the apo-enzyme which is covered by the B-face of the nicotinamide ring when coenzymes are bound . Reduction in the area of this hydrophobic surface results in the mutant tetramer being more thermally stable than the wild-type enzyme.

Nature, 1987 Nov 5-11, 330(6143), 86 - 8
Prediction of electrostatic effects of engineering of protein charges; Sternberg MJ et al.; Accurate prediction of electrostatic effects on catalytic activity is an essential component of protein design . Site-directed mutagenesis of charged groups in subtilisin of Bacillus amyloliquefaciens has provided experimental measurements of electrostatic interactions which may be used to test such theoretical methods . The pKa of the histidine of the active site has been perturbed by +0.08 to -1.0 units by modifying one or two residues . Electrostatic effects in proteins can be modelled by the algorithm of Warwicker and Watson, which uses classical electrostatics and considers both the charge position and the shape of the molecule . Here we report that the algorithm can model several pKa shifts in subtilisin to fair accuracy.

J Clin Microbiol, 1987 Nov, 25(11), 2225 - 6
Nosocomial septicemia with CDC group IV c-2, an unusual gram-negative Bacillus; Crowe HM et al.; A 55-year-old man with severe peripheral vascular disease developed nosocomial septicemia which was caused by the gram-negative bacterium CDC group IV c-2, presumably from a plantar abscess on the left foot . Recovery followed amputation of the infected extremity and antibiotic therapy . This is the first reported case of nosocomial acquisition of this organism.

J Antibiot (Tokyo), 1987 Nov, 40(11), 1506 - 14
New peptide antibiotics LI-F03, F04, F05, F07, and F08, produced by Bacillus polymyxa . I . Isolation and characterization; Kurusu K et al.; A strain of Bacillus polymyxa produced a new peptide antibiotic complex, named LI-F, composed of more than ten components . The components, antibiotics LI-F03, F04, F05, F07, and F08 were isolated from the complex by reversed phase HPLC . They are active against fungi, yeasts, and Gram-positive bacteria . The fast atom bombardment mass spectra revealed that the individual isolated antibiotics are still mixture of two homologous components, being very difficult to separate from each other.

Arch Biochem Biophys, 1987 Nov 1, 258(2), 332 - 41
Permeability of ammonia and amines in Rhodobacter sphaeroides and Bacillus firmus; Ritchie RJ et al.; Permeabilities of uncharged ammonia (NH3), methylamine (CH3NH2), and ethylamine (CH3CH2NH2) in the gram-negative phototrophic bacterium Rhodobacter sphaeroides were measured directly in cells grown heterotrophically under aerobic conditions . The permeability of NH3 was 2.55 +/- 0.73 microns s-1 (n = 20), but the permeabilities of CH3NH2 (MA) and CH3CH2NH2 (EA) were higher, PMA = 17.8 +/- 2.8 microns s-1 (n = 50), PEA = 24.7 +/- 3.9 microns s-1 (n = 44) . The relative permeabilities of amines were also determined from their effect on the pH gradient across the cell membrane at alkaline external pH . In aerobically grown R . sphaeroides, both techniques indicated that the permeability of CH3CH2NH2 was about 30% greater than that of CH3NH2 but that the permeability of NH3 was only about 1/5 that of CH3NH2 . The relative permeabilities of NH3 (A) and CH3NH2 were different in R . sphaeroides cells grown under three different physiological conditions: (a) cells grown aerobically with ammonium sulfate (PA/PMA about 0.20), (b) cells grown anaerobically with ammonium sulfate as their nitrogen source (PA/PMA about 0.29), and (c) diazotrophic cells (PA/PMA about 0.38) . NH3 was also found to be only about 1/3 as permeable as CH3NH2 in the alkalophilic gram-positive bacterium Bacillus firmus . The findings that permeability properties of NH3 and CH3NH2 are very different in different bacteria and vary according to the conditions under which the organism is grown need to be taken into account in the interpretation of experiments where {14C}methylamine is used as an ammonia analog.

Laryngoscope, 1987 Nov, 97(11), 1303 - 6
Tuberculous otitis media: a clinical record; Yaniv E; The clinical picture of tuberculous otitis media has changed since previously documented . In our series of 31 patients, it was found that severe conductive hearing loss, abundant pale granulations, and denuded malleus handle are constant findings and, in our opinion, are significant clinical features of the pathology . The disease can also manifest itself as an acute mastoiditis . As regards to investigations, bacteriology is considered as being unreliable . This is attributed to secondary organisms interfering with the growth of the tubercle bacillus, as well as the fastidious nature of the bacillus itself . We regard histology as the most reliable means of attaining a definitive diagnosis . Treatment was with a four drug antituberculous regime administered over 6 months . Streptomycin was excluded in all but one case due to its ototoxicity . We believe TB otitis media to be secondary to an established chronic otitis media in the majority of cases.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1987 Nov, 20(4), 327 - 38
{Extracellular thermostable alpha-amylase from Bacillus stearothermophilus Q8}; Lin HY et al.; The alpha-amylase of Bacillus stearothermophilus Q8, previously isolated in our laboratory, was purified by ammonium sulfate precipitation and CM-sephadex chromatography . The activity the of partial purified alpha-amylase was to be protected by bovine serum albumin Ca2+ and Mg2+ . The enzyme showed 100% activity at pH 9.0; 98%, at pH 8.0 and 41%; at pH 10.0 . It expressed optimal reaction temperature at 90 degrees C, 81% of the activities remained at 100 degrees C . After 15 min incubation at 100 degrees C with the addition of 10 mM Ca2+, the enzyme only retained 67% activity . The enzyme, however, retained 10% of the maximal activity after 2 h incubation at 90 degrees C, in the absence of substrate and with the addition of Ca2+ . Of cations, Na+ at 0.1 and 1 mM, Mn2+ at 0.1 mM showed stimulatory effect; of anions OH-Cl-I-HCO3-NO2-N3- at 10 mM showed stimulatory effect . Addition of urea and KMnO4 resulted in the loss of enzyme activities; however, lower concentration of SDS and Tween 80 afforded protection of the enzyme activities . Galactose and maltose were non-inhibitory for the enzyme activities, while, fructose, mannose, xylose and lactose were slightly inhibitory . The relative hydrolysis sequence of polysaccharides were amylose greater than soluble starch = corn starch greater than glycogen.

J Comp Pathol, 1987 Nov, 97(6), 677 - 85
The serum amyloid A stimulating factor (SAASF) in the hamster; Hol PR et al.; The acute phase SAA response was studied in hamsters . An SAA-stimulating factor (SAASF) was detected in the early acute phase blood plasma of hamsters which were subcutaneously injected with casein-LPS . The latter is routinely used in our laboratory for amyloid induction in hamsters . Acute (4 h) inflammatory exudates (greater than 80 per cent polymorphonuclear leukocytes) were produced by intraperitoneal injection with either casein-LPS, latex or Freund's incomplete adjuvant . Chronic inflammatory exudate macrophages (greater than 98 per cent) were elicited by intraperitoneal injection with Bacillus Calmette Guerin (BCG) . Cells were stimulated in vitro with latex . SAASF was detected in the supernates and lysates of the acute exudate cells but not in those of the chronic peritoneal exudate macrophages . Lymphocyte activating factor (LAF), however, was evidently present in the latter samples, indicating that SAASF and LAF (IL-1) are functionally different substances in hamsters.

J Gen Microbiol, 1987 Nov, 133 ( Pt 11), 3093 - 7
Genetic mapping of the bacitracin synthetase gene(s) in Bacillus licheniformis; Podlesek Z et al.; The map position of a mutation in the bacitracin synthetase gene(s) in Bacillus licheniformis ATCC 10716 was determined by transduction with phage SP-15 . Results indicate that it is linked to the lys and trp loci and is distinct from the known sporulation loci on the chromosome of Bacillus licheniformis . The defect(s) of the enzyme complex were analysed in terms of its ability to bind covalently 14C-labelled amino acid precursors of the bacitracin molecule.

Biochem Int, 1987 Nov, 15(5), 887 - 95
Reconstitution and crystallisation experiments with isolated split proteins from Bacillus stearothermophilus ribosomes; Gewitz HS et al.; Six proteins (B-L1, B-L6, B-L10, B-L11, B-L12 and B-L16) were removed from 50S ribosomal subunits of Bacillus stearothermophilus by treatment with ethanol and ammonium chloride . The proteins were isolated in a pure form, and one of them (B-L6) was crystallized . Five of the six proteins (in various combinations) were added back to the core particles, resulting in 50S subunits lacking one protein . The biological activities of these ribosomal particles as determined in the poly(U)-system varied over a wide range, depending on the protein which was omitted . The particles lacking one protein provide useful tools for heavy-atom derivation necessary for our crystallographic studies on the 50S subunits of Bacillus stearothermophilus.

Carbohydr Res, 1987 Nov 1, 168(2), 285 - 94
Formation of 6-O-alpha-maltosylcyclomalto-oligosaccharides by transfer action of three debranching enzymes; Yoshimura Y et al.; O-Maltosylcyclomaltohexaoses (G2-cG6) were formed in yields of 24.3 and 23.2 mmol from 40 mmol of alpha-maltosyl fluoride (alpha-G2F) and 90 mmol of cyclomaltohexaose (cG6) by the transfer action of pullulanase from Aerobacter aerogenes (A-pullulanase) and isoamylase from Pseudomonas amyloderamosa, respectively . These yields were three times that given by pullulanase from Bacillus acidopullulyticus (B-pullulanase) . The yields of O-maltosylcyclomalto-oligosaccharides were changed according to the origin of the enzymes and the kind of cyclomalto-oligosaccharide (cG6, cG7, or cG8) used as the acceptor . By the reaction with 40 mmol of alpha-G2F and 90 mmol of cG6, 20 mmol of alpha-G2F and 30 mmol of cG7, or 40 mmol of alpha-G2F and 90 mmol of cG8, the amounts of O-maltosylcyclomalto-oligosaccharides produced and the transfer ratios of alpha-G2F to the acceptors were as follows . By A-pullulanase, 24.3 mmol of G2-cG6 was produced in a 60.8% transfer ratio, whereas the yields of G2-cG7 and G2-cG8 were 1.7 mmol (8.5%) and 8.4 mmol (21.0%), respectively . The yields of G2-cG6, G2-cG7, and G2-cG8 by B-pullulanase were 8.8 mmol (22.0%), 1.2 mmol (6.0%), and 11.7 mmol (29.3%), respectively . In the case of isoamylase, G2-cG7 (9.2 mmol, 46.0%) and G2-cG8 (20.9 mmol, 52.3%) were produced, as much as for G2-cG6 (23.2 mmol, 58.0%) . It was suggested that the difference in the amounts of G2-cG6 produced by these three debranching enzymes is based on the difference in the mode of action on the alpha-G2F used as the substrate, either a transfer action or a hydrolytic action.

Carbohydr Res, 1987 Nov 1, 168(2), 211 - 8
Isolation and structure determination of a diacetamidodideoxyuronic acid-containing glycan chain from the S-layer glycoprotein of Bacillus stearothermophilus NRS 2004/3a; Messner P et al.; A glycan isolated from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a was shown by 1H- and 13C-n.m.r . spectroscopy to have the tetrasaccharide repeating-unit ----4)-beta-ManpA2,3(NAc)2-(1----3)-alpha-GlcpNAc-(1----4)-beta- ManpA2,3(NAc)-(1----6)-alpha-Glcp(1----.

Ann Gastroenterol Hepatol (Paris), 1987 Nov, 23(6), 347 - 9
{Segmental tuberculosis of the colon manifested by rectal stenosis}; Rodier B et al.; Colonic tuberculosis is currently exceptionally rare in western countries . This interesting report reminds of its existence and emphasizes the diagnostic problems with Crohn's disease in the past . The barium enema enables to show the main aspects of colonic tuberculosis, i.e . variable forms of stenosis . The contribution of colonoscopy is essential in defining the lesions and allowing multiple biopsies . These biopsies may show the presence of granulomas with caseous necrosis, and possibly the Koch Bacillus on direct examination or in culture . The conditions of prescription of the medical treatment are similar to that of other tuberculous locations . Surgery is indicated in the presence of complications, when there is a doubt the malignant nature of the lesion, finally at the stage of cicatricial stenosis.

Mikrobiologiia, 1987 Nov-Dec, 56(6), 947 - 50
{Fibrinolytic activity of natural variants of Bacillus mesentericus}; Imshenetskii AA et al.; The natural variability of the ability to synthesize proteinases by Bacillus mesentericus 64 was studied . The population of this strain was shown to be heterogeneous . Three types of variants (S, M and P) differed in the morphology of their colonies and in the culture characteristics from the typical colonies of the parent strain . The caseinolytic activity of the M variant was three times as high as that of the parent strain, and it also had an elevated fibrinolytic activity and a high rate of blood thrombolysis in experiments in vitro . The rate of proteinase synthesis correlated with the morphological types of sporogenic bacteria.

J Appl Bacteriol, 1987 Nov, 63(5), 449 - 53
Effect of the method of preparation of bacille Calmette-Guérin (BCG) vaccine on the properties of four daughter strains; Abou-Zeid C et al.; Samples of Bacille Calmette-Guerin (BCG) vaccines from four collaborating production laboratories, each of which had prepared vaccine from four different daughter strains of BCG, had previously been monitored for changes in colony morphology and the present study was undertaken to determine whether the changes observed were reflected in the patterns of protein secretion and lipid content . In the samples examined there was evidence for a correlation between colony morphology and the presence or absence of mycoside B . As the components of BCG that determine virulence and protective immunity are unknown, care must be taken to ensure constancy of the strains during the manufacture of vaccines.

J Chir (Paris), 1987 Nov, 124(11), 612 - 4
{Tuberculosis of the hip: diagnostic problems . Apropos of 44 cases}; Meziane A et al.; Diagnostic difficulties in 44 cases of tuberculosis of hip are emphasized and the serious nature of sequelae stressed . A definite diagnosis (cytology positive for Kochs bacillus, positive biopsy) was possible in only 18 cases, presumption of the nature of the affection in the other cases depending on clinical, radiologic and biologic arguments and failure to respond to immobilization one month.

Appl Environ Microbiol, 1987 Nov, 53(11), 2680 - 2
Evaluation of the Minitek system for characterization of Bacillus species; Sullivan NM et al.; Minitek (BBL Microbiology Systems, Cockeysville, Md.) substrate disks were evaluated as alternatives to conventional tests for the characterization of Bacillus species . Results were compared for 10 reference isolates and 87 isolates from food sources . The overall agreement of results between the Minitek and conventional tests was 92% for reference strains and 86% for food isolates.

Infect Immun, 1987 Nov, 55(11), 2860 - 3
Most Mycobacterium leprae carbohydrate-reactive monoclonal antibodies are directed to lipoarabinomannan; Gaylord H et al.; Each of more than 30 monoclonal antibodies that had been raised against Mycobacterium leprae and previously classified as reactive with carbohydrate was shown to be directed against lipoarabinomannan, a prominent, highly pervasive, myo-inositol-phosphate-containing, cross-reactive antigen within the leprosy bacillus . Some of the antibodies preferentially bound to the lipopolysaccharide of M . leprae rather than to that of Mycobacterium tuberculosis, suggesting the presence of distinguishing structural features . The presence of alkali-labile inositol 1-phosphate in the lipopolysaccharide from M . tuberculosis and its apparent absence from the M . leprae product may account for the difference.

Mikrobiologiia, 1987 Nov-Dec, 56(6), 956 - 62
{Effect of initiated spores on the resistance of nongerminated resting forms of Bacillus cereus remaining in the suspension to the action of damaging agents}; Pronin SV; The elevated resistance of a Bacillus cereus spore suspension against the action of UV was found to depend on the quantity of resting forms initiated in the suspension prior to an irradiation . The resistance against UV increased 80-50 times if 60-90% of spores were initiated in the suspension as compared to that of the original resting forms . When suspensions containing 40% of non-germinated B . cereus spores were kept at 4 degrees C for 14 days, the latter became 10 and 14 times more resistant to elevated temperature (90 degrees C) and chloramine (2.5%), respectively, as compared to control intact spores . The higher resistance of non-germinated spores against the action of physical and chemical damaging agents was registered within the entire period of experiments (over three months) . This phenomenon was not observed if ca . 100% of spores were initiated in a suspension . The resistance of initiated spores against the action of UV was 40 times lower than that of B . cereus resting forms.

Can J Microbiol, 1987 Nov, 33(11), 982 - 9
Binding of the Bacillus sphaericus mosquito larvicidal toxin to cultured insect cells; Davidson EW et al.; Using both fluorescent labelled toxin and antibody--secondary antibody techniques, the Bacillus sphaericus toxin was found to bind strongly to susceptible Culex quinquefasciatus cells, but far less strongly to cells of insensitive insects . An insensitive clone of the C . quinquefasciatus cell line was discovered which bound toxin efficiently . The toxin was bound in the cold to sensitive cells and these cells could be rescued from cytotoxicity for ca . 15 min after warming, by which time toxin appeared to be internalized . Binding was saturable . This toxin is apparently internalized by receptor-mediated endocytosis, probably involving a glycoprotein receptor containing N-acetyl-D-glucosamine . Evidence for toxin binding to lipids was not found . Antibody appeared to detect internalized toxin, and high concentrations of sugars inhibited cytotoxicity; these results along with evidence from a recent ultrastructural study suggest that this toxin may form pores in the cell membrane.

Prostaglandins, 1987 Nov, 34(5), 633 - 42
Increased arachidonic acid metabolites from cells in culture after treatment with the phosphatidylcholine-hydrolyzing phospholipase C from Bacillus cereus; Levine L et al.; Treatment of rat liver cells (the C-9 cell line), porcine aorta endothelial cells, bovine aorta smooth muscle cells, bovine aorta endothelial cells, mouse fibroblasts and rat keratinocytes with highly purified, crystallized Bacillus cereus phospholipase C, which hydrolyzes phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine but has little or no effect on phosphatidylinositol, phosphatidylglycerol, cardiolipin, sphingomyelin, lysophosphatidylcholine or lysophosphatidylethanolamine, increased metabolism of arachidonic acid . Hydrolysis of phosphatidylcholine (and/or phosphatidylethanolamine) by a phosphatidylcholine (or phosphatidylethanolamine)-hydrolyzing phospholipase C appears to contribute to liberation of substrate for arachidonic acid metabolism.

J Infect, 1987 Nov, 15(3), 207 - 12
Toxigenic Bacillus cereus as a cause of wound infections in the tropics; Dryden MS et al.; A bacteriological survey was conducted on clinically infected traumatic wounds in members of an Operation Raleigh expedition, who were working in the Costa Rican rain forest . Bacillus cereus was isolated from the wounds of 14 of 18 patients, usually in pure and heavy growth . Most of the isolates were strongly toxigenic by in vivo pathogenicity tests . The organism was also isolated from the nose in 15 cases and the throat in five cases . The findings indicate that B . cereus was the principle pathogen in this series of traumatic wounds.

J Leukoc Biol, 1987 Nov, 42(5), 498 - 503
1 alpha,25 Dihydroxyvitamin D3 and mononuclear phagocytes: enhancement of mouse macrophage and human monocyte hydrogen peroxide production without alteration of tumor cytolysis; Gluck WL et al.; 1 alpha,25 dihydroxyvitamin D3 (1,25 D3) is known to interact with hematopoietic cells . The purpose of this study was to determine the effect of 1,25 D3 on hydrogen peroxide (H2O2) production and tumor cell killing by mouse peritoneal macrophages and human blood monocytes . Enhanced monocyte and macrophages phorbol myristate acetate (PMA)-stimulated H2O2 production was observed at concentrations of 0.13 to 130 nM 1,25 D3 and and was maximal at 1.3 nM . At concentrations of 100 U/ml, gamma interferon (IFN-gamma) alone had a similar effect but, in combination with 1,25 D3, there was no cooperative effect . At concentrations ranging from 0.13 to 130 nM, 1,25 D3 failed to augment tumor cell lysis by macrophages from peptone-injected normal or bacillus Calmette-Guerin (BCG)-infected mice, or by blood monocytes from normal humans . Our results indicate that 1,25 D3 can activate the monocyte and macrophage for H2O2 secretion without concomitant activation for tumor cell killing.

J Urol, 1987 Nov, 138(5), 1158 - 61
5-year experience with intravesical therapy of carcinoma in situ: an inquiry into the risks of "conservative" management; Stanisic TH et al.; Recent reports of disease progression in patients being treated with intravesical agents for carcinoma in situ of the bladder led us to examine our 5-year experience with 26 consecutive patients with carcinoma in situ treated with intravesical therapy for this lesion . Nine patients with isolated carcinoma in situ and 17 with carcinoma in situ associated with papillary lesions were treated intravesically with a variety of agents in a closely monitored program for a mean of 22 months . All patients wished to exhaust conservative options before accepting cystectomy . Of 26 patients treated 9 (35 per cent) have no evidence of disease with a functional bladder after 27 months, 13 (50 per cent) are treatment failures and 4 (15 per cent) maintain an equivocal status to date . Successful treatment results with each drug were 6 of 24 thiotepa, 0 of 7 mitomycin, 0 of 6 doxorubicin and 3 of 8 bacillus Calmette-Guerin . Treatment failure was associated with disease progression to muscle invasion, adjacent organ invasion or metastases in 7 patients (27 per cent) . Significant reduction in bladder capacity after prolonged therapy occurred in 3 patients . Our results suggest that persistent, intensive, "conservative" intravesical therapy in the face of recurrent or persistent disease is associated with substantial patient risk.

Cancer Res, 1987 Nov 1, 47(21), 5590 - 4
Synthesis of nitrite and nitrate in murine macrophage cell lines; Stuehr DJ et al.; Synthesis of nitrite (NO2-) and nitrate (NO3-) was studied in the macrophage cell lines RAW 264.7, WEHI-3, PU5-1.8, J774A.1, and P388D1 and compared to the synthesis by thioglycolate-elicited peritoneal macrophages from C3H/He and C3H/HeJ mice . Treatment with Escherichia coli lipopolysaccharide (LPS) induced NO2-/NO3- synthesis by all the cell lines except P388D1, which remained unresponsive at the highest LPS concentration (50 micrograms/ml) . Recombinant murine gamma-interferon induced NO2-/NO3- synthesis in only two cell lines (PU5-1.8 and RAW 264.7), although it activated synthesis by C3H/He and C3H/HeJ macrophages . Dual signal treatments consisting of lymphokines or gamma-interferon plus LPS stimulated NO2-/NO3- synthesis by all five cell lines and each line showed enhanced synthesis as compared to that induced by any single stimulus . Heat-killed Bacillus Calmette-Guerin and purified mycobacterial protein derivative stimulated NO2-/NO3- synthesis in three of five cell lines, while dextran sulfate, zymosan, and the synthetic adjuvant muramyl dipeptide were ineffective . Nitrite represented 50-75% of the total NO2-/NO3- produced in all cases . The kinetics of LPS-induced NO2-/NO3- synthesis in J774A.1 and C3H/He macrophages were identical; a 6-h lag phase was followed by a 24- to 48-h period in which NO2- and NO3- were in a ratio of approximately 3:2 at all time points.

J Infect Dis, 1987 Nov, 156(5), 763 - 9
Mycobacterial carbohydrate antigens for serological testing of patients with leprosy; Levis WR et al.; To determine whether quantitation of antibodies to mycobacterial carbohydrate determinants would be valuable in serodiagnosis and monitoring of leprosy, we tested serum IgM antibody to Mycobacterium leprae phenolic glycolipid I and IgM and IgG antibodies to Mycobacterium tuberculosis and M . leprae lipoarabinomannan (LAM) by enzyme-linked immunosorbent assay . Seventy-one percent of patients with paucibacillary disease and 85.5% of patients with multibacillary disease were positive for at least one of the three antibodies . The 15% of antibody-negative patients with multibacillary disease were mostly long-term-treated patients, with inactive disease by biopsy . There was excellent agreement between M . tuberculosis LAM and M . leprae LAM in detection of antibodies . Bacillary index and levels of both IgG and IgM antibodies to LAM were positively correlated when all patients were analyzed . When patients with a history of erythema nodosum leprosum (ENL) were analyzed separately, there was no correlation between IgM or IgG antibody to LAM and bacillary index, a result suggesting a possible role for LAM in the pathogenesis of ENL.

Transplantation, 1987 Nov, 44(5), 621 - 32
Specific tolerance and immunocompetence in haploidentical, but not in completely allogeneic, canine chimeras treated with methotrexate and cyclosporine; Deeg HJ et al.; Recipient dogs were conditioned with 9.2 Gy of total-body irradiation followed by the infusion of bone marrow and peripheral blood leukocytes from a DLA-haploidentical littermate (N = 10) or a completely allogeneic unrelated donor (n = 9) . Graft-vs.-host disease (GVHD) prophylaxis consisted of methotrexate (MTX) and cyclosporine (CsA) . Postgrafting all dogs were complete lymphohemopoietic chimeras . Lymphocytes of haploidentical chimeras without GVHD were unresponsive to stimulation by host lymphocytes cryopreserved pregrafting . Lymphocytes of haploidentical chimeras with GVHD proliferated in response to host cells, albeit less than donor cells pregrafting . In completely allogeneic chimeras, neither lymphocytes from dogs with GVHD, nor those from dogs without the disease showed responses to host lymphocytes . In addition, cells from haploidentical chimeras obtained early after transplantation nonspecifically suppressed donor cell proliferation . Later on, lymphocytes from dogs without GVHD showed specific suppression of donor cells, while lymphocytes from chimeras with GVHD continued to show nonspecific suppression . Cells from completely allogeneic chimeras both with and without GVHD never suppressed donor cells specifically . Both specific and nonspecific suppressor cells were enriched by nylon wool adherence, expressed T cell markers, and were not affected by the addition of indomethacin . Even after removing nylon wool-adherent cells, however, chimera cells were unresponsive to stimulation by host cells . By one year after transplant, chimera lymphocytes no longer showed suppression . In cell-mediated lympholysis assays, lymphocytes from all chimeras, regardless of GVHD, failed to generate cytotoxic cells against host cell targets . However, while haploidentical chimeras showed cytotoxicity against third-party targets, completely allogeneic chimeras did not . This deficiency was not overcome by the addition of mixed leukocyte culture supernatant or donor lymphocytes . All chimeras had basically normal antibody responses to keyhole limpet hemocyanin and phage X174 . However, while haploidentical chimeras had normal responses to bacillus Calnette-Guerin (BCG) sensitization and rejected DLA-incompatible skin grafts within the normal time frame, completely allogeneic chimeras were not sensitized by BCG and showed delayed skin graft rejection . Histopathological studies revealed slow thymic reconstitution in all chimeras, particularly in the presence of GVHD . However, while healthy haploidentical chimeras eventually showed thymic histology normal for age, completely allogeneic chimeras did not.(ABSTRACT TRUNCATED AT 400 WORDS)

Mol Microbiol, 1987 Nov, 1(3), 283 - 91
The use of DNA probes identifying restriction-fragment-length polymorphisms to examine the Mycobacterium avium complex; McFadden JJ et al.; DNA probes were used to identify restriction-fragment-length polymorphisms (RFLPs) in DNA samples, demonstrating that the Mycobacterium avium complex could be clearly divided into M . avium and Mycobacterium intracellulare strains . Less than 2% DNA base substitution was found between M . avium strains, whereas the M . intracellulare strains had greater than 15% base substitution . The Johne's disease bacillus, Mycobacterium paratuberculosis (American type strain), was found to be distinguishable from the M . avium complex serotypes examined . Strain 18 was found to be identical to M . avium . The rat leprosy bacillus, Mycobacterium lepraemurium, was found to be very closely related, but not identical, to M . avium.

Mol Gen Mikrobiol Virusol, 1987 Nov, (11), 23 - 7
{The role of plasmids in the regulation of delta-endotoxin synthesis in Bacillus thuringiensis H14}; Stepanova TV et al.; The role of plasmids in regulation of delta-endotoxin synthesis by Bacillus thuringiensis H14 was studied . The derivatives of strain Is-1 H14 containing a 4Md plasmid integrated into the chromosome synthesize small crystals and are not toxic for the gnat larvae . The transceptional transfer into this strain of a plasmid coding for crystal synthesis from the strain 69-6 serotype H5 results in restoration of insecticidal activity to the level of the parental strain Is-1 . Transcipients activity is increased 10-15 fold in case of 4Md plasmid excision from the chromosome and autonomous functioning . Evidently, 4Md plasmid from the strain Is-1 as well as a plasmid coding for crystal synthesis from the strain 69-6 contains the regulatory elements participating in the expression of crystalline protein genes localized on other plasmids . The existence of two cellular regulatory groups is supposed to result in the significant increase in crystalline protein synthesis.

Biochem Int, 1987 Nov, 15(5), 1057 - 67
Nucleotide sequence and linkage map position of the secX gene in maize chloroplast and evidence that it encodes a protein belonging to the 50S ribosomal subunit; Markmann-Mulisch U et al.; The nucleotide sequence of the segment of maize chloroplast DNA lying between the map coordinate positions 32.59 and 32.98 Kb and containing the secX gene has been determined . The derived amino acid sequence of maize chloroplast secX is 95%, 87% and 62% identical to the corresponding derived amino acid sequences from two plant chloroplasts and Escherichia coli, respectively . It is also 70% identical to the experimentally determined amino acid sequence of a protein isolated from Bacillus stearothermophilus ribosomes . Separation of the 50S ribosomal subunit proteins of E . coli by reversed phase HPLC gave a peak which contained pure secX protein, as determined by N-terminal amino acid sequencing . Spinach chloroplast 50S subunit proteins separated by HPLC also gave a peak corresponding to pure secX protein . From these results we conclude that the secX gene in E . coli and in plant chloroplasts encodes a small (37-38 amino acid residues) ribosomal protein belonging to the 50S subunit . The same conclusion has been reached recently by A . Wada with respect to E . coli secX . In agreement with Wada, we name the secX protein L36 . Its chloroplast gene is designated rpL36.

Appl Environ Microbiol, 1987 Nov, 53(11), 2656 - 9
Molecular cloning of an endoglucanase gene from an alkalophilic Bacillus sp . and its expression in Escherichia coli; Kim JM et al.; One of the cellulase genes from alkalophilic Bacillus sp . strain N-4 was cloned in pBR322 . A recombinant plasmid, pYBC107, expressing carboxymethyl cellulase (CMCase) was isolated, and the size of the cloned HindIII fragment was found to be 5.5 kilobases . The restriction map of pYBC107 showed a different pattern from those of pNKI and pNKII (N . Sashihara, T . Kudo, and K . Horikoshi, J . Bacteriol . 158:503-506, 1984) . When the HindIII fragment from pYBC107 was subcloned into pYEJ001, there was a 3.8-fold increase in CMCase activity over that observed with pYBC107 . Plasmid pYBC108 constructed by treatment of pYBC107 with HindIII and EcoRI expressed the CMCase activity, although to a limited extent . To verify the originality of cloned pYBC107 from Bacillus sp., we analyzed the restriction digest by Southern blotting.

Appl Environ Microbiol, 1987 Nov, 53(11), 2650 - 5
The glycoprotein toxin of Bacillus thuringiensis subsp . israelensis indicates a lectinlike receptor in the larval mosquito gut; Muthukumar G et al.; The mosquito-active protein crystals produced by Bacillus thuringiensis subsp . israelensis contain covalently attached aminosugars which are critical for their larvicidal activity . The 50% lethal concentrations toward Aedes aegypti larvae were increased up to 10-fold by mild periodate treatment, up to 40-fold by forming the protein crystals in the presence of tunicamycin, and up to 7-fold by the presence during the mosquito bioassays of N-acetylglucosamine or its trimer, triacetylchitotriose . Periodate-treated crystals and crystals formed in the presence of tunicamycin had greatly reduced binding capacities for wheat germ agglutinin, an N-acetylglucosamine-specific lectin . These results suggest that the B . thuringiensis subsp . israelensis glycoprotein toxin binds to a lectinlike receptor in the larval mosquito gut . Furthermore, the distinct lectin-binding patterns exhibited by diptera-active versus lepidoptera-active B . thuringiensis crystals suggest that host specificity for the microbial insecticides is determined, in part, by the carbohydrate portion of their glycoprotein crystals.

Biochemistry, 1987 Oct 20, 26(21), 6621 - 6
Solid-state 15N NMR of oriented lipid bilayer bound gramicidin A'; Nicholson LK et al.; Highly oriented samples of lipid and gramicidin A' (8:1 molar ratio) have been prepared with the samples extensively hydrated (approximately 70% water v/w) . These preparations have been shown to be completely in a bilayer phase with a transition temperature of 28 degrees C, and evidence is presented indicating that the gramicidin is in the channel conformation . An estimate of the disorder in the alignment of the bilayers parallel with the glass plates used to align the bilayers can be made from the asymmetry of the nuclear magnetic resonances (NMR) . Such an analysis indicates a maximal range of disorder of +/- 3 degrees . Uniformly 15N-labeled gramicidin has been biosynthesized by Bacillus brevis grown in a media containing 15N-labeled Escherichia coli cells as the only nitrogen source . When prepared with labeled gramicidin, the oriented samples result in high-resolution 15N NMR spectra showing 12 resonances for the 20 nitrogen sites of the polypeptide . The frequency of the three major multiple resonance peaks has been interpreted to yield the approximate orientation of the N-H bonds in the peptide linkages with respect to the magnetic field . These bond orientations are only partially consistent with the extant structural models of gramicidin.

Am J Ophthalmol, 1987 Oct 15, 104(4), 325 - 33
Microbial contamination of contact lens care systems; Donzis PB et al.; We examined the contact lens care systems of 100 asymptomatic patients who used hard or soft contact lenses for correction of refractive errors for the presence of bacteria, fungi, Acanthamoeba, and endotoxin . Of 100 patients, 52 had contaminated contact lens care systems . Sixteen of 126 bottles (13%) of commercial contact lens care solutions were contaminated . Contaminated commercial solutions were opened and used for a longer period of time than uncontaminated solutions . Contamination was not found in bottles of preserved commercial solutions that were opened and used for less than 21 days . All 12 bottles of homemade saline were contaminated with bacteria, and Acanthamoeba was isolated from two of these bottles . Pseudomonas was found in the care systems of 12 patients . Bacillus species, which form spores resistant to heat, were found in the care systems of seven patients . Endotoxin, which is also resistant to heat, was detected in nine of 35 care systems (26%) tested . Potential pathogens were isolated from the conjunctiva of six patients.

J Biol Chem, 1987 Oct 15, 262(29), 14343 - 51
The primary structure of rat ribosomal protein S12 . The relationship of rat S12 to other ribosomal proteins and a correlation of the amino acid sequences of rat and yeast ribosomal proteins; Lin A et al.; The covalent structure of the rat 40 S ribosomal subunit protein S12 was determined from the sequence of amino acids in tryptic, chymotryptic, thermolytic, and cyanogen bromide peptides and inferred from the sequence of nucleotides in a recombinant cDNA . Rat ribosomal protein S12 contains 129 amino acids and has a molecular weight of 14,120 . The amino acid sequences of a number of ribosomal proteins appear to be related to rat S12 . These include spinach chloroplast L7, Escherichia coli S5, Nicotiana tabacum chloroplast S18, and Bacillus stearothermophilus S12, and perhaps others . In addition, there are two sequences, 26 and 18 amino acids in length, in rat S12 that may be related to segments of the same number of residues in ribosomal proteins from a number of species . These, and other results, reinforce the suggestion that ribosomal proteins form an extended family.

Biochem Biophys Res Commun, 1987 Oct 14, 148(1), 515 - 20
Synthesis of 4R- and 4S-tritium labeled NADPH for the determination of the coenzyme stereospecificity of NADPH: protochlorophyllide oxidoreductase; Valera V et al.; A rapid and easy method for the production of both the 4R and 4S tritium labeled isomers of either NADH or NADPH has been developed . The method requires the use of only a single labeled compound (D-{1(-3)H} glucose), and two enzymes (glucose dehydrogenase from Bacillus sp . and alcohol dehydrogenase from Thermoanaerobium brockii) which are specific for the pro S and pro R hydrogens, respectively, of either NADH or NADPH . The 4R and 4S tritium labeled isomers of NADPH have been used to determine that NADPH:protochlorophyllide oxidoreductase from etiolated wheat was specific for the pro S hydrogen of NADPH.

Biochem Biophys Res Commun, 1987 Oct 14, 148(1), 15 - 23
Rational construction of a 2-hydroxyacid dehydrogenase with new substrate specificity; Clarke AR et al.; Using site-directed mutagenesis on the lactate dehydrogenase gene from Bacillus stearothermophilus, three amino acid substitutions have been made at sites in the enzyme which we suggest in part determine specificity toward different hydroxyacids (R-CHOH-COOH) . To change the preferred substrates from the pyruvate/lactate pair (R = -CH3) to the oxaloacetate/malate pair (R = -CH2-COO-), the volume of the active site was increased (thr 246----gly), an acid was neutralized (asp-197----asn) and a base was introduced (gln-102 - greater than arg) . The wild type enzyme has a catalytic specificity for pyruvate over oxaloacetate of 1000 whereas the triple mutant has a specificity for oxaloacetate over pyruvate of 500 . Despite the severity and extent of these active site alterations, the malate dehydrogenase so produced retains a reasonably fast catalytic rate constant (20 s-1 for oxaloacetate reduction) and is still allosterically controlled by fructose-1,6-bisphosphate.

J Antibiot (Tokyo), 1987 Oct, 40(10), 1431 - 9
Metabolic products of microorganisms . 239 . Bacimethrin isolated from Streptomyces albus identification, derivatives, synthesis and biological properties; Drautz H et al.; Bacimethrin (1), known as a thiamine antagonist produced by Bacillus megatherium, was isolated from Streptomyces albus and has been further characterized by NMR spectra and acetylation . A new easy three step synthesis for 1 is described . The biological activity of 1, and its mode of action were discussed . There are indications that bacimethrin inhibits the phosphorylation of 4-amino-5-hydroxymethyl-2-methylpyrimidine (Pyr-OH) during thiamine biosynthesis.

Arch Intern Med, 1987 Oct, 147(10), 1815 - 6
Regression of melanoma nodules in a patient treated with ranitidine; Smith T et al.; Human malignant melanoma may regress spontaneously or with immunotherapy, such as Calmette-Guerin bacillus, interferon alfa, interleukin-2, and interleukin-2 plus lymphokine-activated killer cells . Histamine type 2 receptor antagonists can modulate immune function by inhibiting suppressor T-cell induction and activity, and melanoma regressions have been reported after the use of cimetidine with coumarin or interferon alfa . This article describes the complete regression of melanoma nodules in a patient treated with ranitidine hydrochloride, another histamine type 2-receptor antagonist . Ranitidine and cimetidine should be considered to be possibly active immunotherapeutic agents in the design and evaluation of clinical trials.

J Urol, 1987 Oct, 138(4), 766 - 70
The outcome of conservative treatment of carcinoma in situ of the bladder; Prout GR Jr et al.; We treated 52 patients with carcinoma in situ by transurethral resection, thiotepa and other intravesical chemotherapeutic agents . All patients underwent standard initial and subsequent evaluative procedures and the average followup was 62 months . Half of the patients had a history of stage Ta and/or T1 transitional cell carcinoma . The remainder had carcinoma in situ when first diagnosed (10 had carcinoma in situ only) . Of 12 patients treated by transurethral resection alone 1 reached 60 months without radical cystectomy or disease progression . There were 18 patients who had a complete response following chemotherapy, 11 had a partial response (positive cytology) and 11 failed (persistent carcinoma in situ) . Patients with a history of transitional cell carcinoma had a statistically significantly greater probability of achieving a complete response . Despite other types of treatments only 2 of 22 patients (partial response and failure) achieved a lasting complete response . Persistent partial response and failure resulted in progressive transitional cell carcinoma (stage T2 or greater, prostatic involvement and metastases) and only 1 of these survived for more than 5 years without cystectomy . None of our patients received bacillus Calmette-Guerin because it was not available during the time most of the patients were treated . While the lives and bladders in some patients may be spared by its use, failure to achieve a complete response indicates impending disaster and cystectomy should be considered seriously.

J Clin Oncol, 1987 Oct, 5(10), 1523 - 33
Chemotherapy versus chemoimmunotherapy (CAF v CAFVP v CMF each +/- MER) for metastatic carcinoma of the breast: a CALGB study . Cancer and Leukemia Group B; Aisner J et al.; Three combination chemotherapy regimens each with or without the methanol-extracted residue of bacillus Calmette-Guerin (BCG) (MER) were compared for efficacy . After stratification for disease-free interval and dominant sites of disease, patients were randomized to either CMF (cyclophosphamide {CYC}, 100 mg/m2 orally, days 1 through 14; methotrexate {MTX}, 40 mg intravenously {IV}, days 1 and 8; 5-fluorouracil {5-FU}, 500 mg/m2 IV, days 1 and 8), or CAF (CYC, 100 mg/m2 orally, days 1 through 14; doxorubicin {DOX}, 25 mg/m2 IV, days 1 and 8; 5-FU, 500 mg/m2 IV, days 1 and 8), or CAFVP (CAF as above plus vincristine {VCR}, 1.0 mg/m2 IV, days 1 and 8; and prednisone {PRED}, 40 mg/m2 orally, days 1 through 14) . Nonspecific immunotherapy with MER was administered in five sites at 100 micrograms or at the lowest tenfold dilution that produced a 1-cm indurated lesion . A total of 432 patients were entered, but 37 were disqualified, leaving 395 evaluable for treatment results and toxicities . One hundred thirty-five evaluable patients were randomized to chemoimmunotherapy until October 28, 1978 . One hundred twenty-six evaluable patients were randomized to chemotherapy alone in the same time period . For the entire study, a total of 260 evaluable patients were randomized to chemotherapy . Chemoimmunotherapy patients were compared with the initial 126 chemotherapy patients . Chemotherapy regimens were compared among all 260 patients . Patient characteristics were similar between regimens and between chemotherapy and chemoimmunotherapy treatment groups . For patients on chemotherapy plus MER, there was no significant differences between the regimens for response frequencies: 43%, 41%, and 32%, respectively for CMF, CAF, and CAFVP . The comparable chemotherapy alone group had 36%, 58%, and 63% response, respectively . The response rates, adjusted for chemotherapy regimen, were 52% and 38% (P = .02) for chemotherapy and chemoimmunotherapy, respectively . MER was associated with painful ulcers and fevers . Thus, MER produced toxicity without response or survival benefit and further randomization after October 28, 1978 was to chemotherapy alone . For 260 evaluable patients on chemotherapy alone, the complete (CR) and partial responses (PR) were 37%, 55%, and 58%, respectively for CMF, CAF, and CAFVP . These response rates for CAF and CAFVP were significantly better than CMF (P = .01 and P less than .01, respectively) . These comparisons were consistent within subgroupings such as dominant sites of disease.(ABSTRACT TRUNCATED AT 400 WORDS)

J Dairy Sci, 1987 Oct, 70(10), 2032 - 9
Preparation and characterization of immobilized beta-lactamase for destruction of penicillin in milk; Lee MZ et al.; beta-Lactamase I (Bacillus cereus) was covalently bound to cyanogen bromide-activated, crosslinked agarose . An initial 5.00 mg of soluble beta-lactamase were used in the immobilization reaction for each preparation, and average coupling yield was 80.5% . Of the enzyme immobilized on the matrix, an average 53.4% remained active . To minimize diffusional effects on immobilized enzyme activity, reaction mixtures were rotated at 250 rpm throughout the study . The shape of the pH activity curve of the immobilized enzyme was identical to that of the soluble enzyme; both exhibited optimum pH around 7.0 . In general, only 2-fold differences in Michaelis constant and maximum volume were observed between native and immobilized enzyme when penicillin G was used as the substrate . However, the Michaelis constant of the immobilized enzyme increased up to 22-fold that of the native enzyme when cephaloridine was used as the substrate . The immobilized enzyme exhibited enhanced stability in the acidic pH region in contrast to the native enzyme, which had superior stability in the alkaline pH region . The heat stability of the immobilized enzyme was about twice that of native enzyme after heat treatment at 60 degrees C for 30 min . Approximately a 10% increase of storage stability on immobilization of beta-lactamase was observed when stored at room temperature (23 +/- 1 degree C) for up to 6 d in the absence of antimicrobial agents . Little loss of activity (less than 2%) was noted after repeated use of the immobilized enzyme up to seven times each in 10.0 ml of skim milk containing .5 U/ml penicillin G.(ABSTRACT TRUNCATED AT 250 WORDS)

J Clin Lab Immunol, 1987 Oct, 24(2), 81 - 5
Reduced in vitro tuberculin reactivity of lymphocytes from patients with tuberculosis; Onwubalili JK; 29 patients with newly diagnosed tuberculosis, and matched healthy controls, were studied on the basis of 3H-thymidine incorporation of peripheral blood mononuclear cells (PBMC) induced in vitro by staphylococcal enterotoxin A plus tetrahydrophorbyl acetate (SEA+TPA) or tuberculin purified protein derivative (PPD) . PBMC from patients had normal SEA+TPA-induced, but depressed spontaneous (p less than 0.01) and PPD-induced (p less than 0.01) 3H-thymidine uptake . Their peak responses tended to occur later (p = 0.007) and with larger doses of the antigen (p = 0.02) . The group of patients with low in vitro PBMC responses to PPD were not clinically distinguishable with respect to the extent of pulmonary tuberculosis, cutaneous reactivity to PPD, nutritional status, bacillary content of sputum or time to sputum sterilization during treatment . Evidence for a plasma factor blocking lymphocyte proliferation was not found in 15 patients tested . Chemotherapy was associated with restoration of normal PPD-induced 3H-thymidine uptake, concomitant with clinical improvement and recovery of nutritional abnormalities.

J Biochem (Tokyo), 1987 Oct, 102(4), 803 - 11
Properties of bovine erythrocyte acetylcholinesterase solubilized by phosphatidylinositol-specific phospholipase C1; Taguchi R et al.; The properties of acetylcholinesterase solubilized from bovine erythrocyte membrane by phosphatidylinositol (PI)-specific phospholipase C of Bacillus thuringiensis or with a detergent, Lubrol-PX, were studied . The activity of Lubrol-PX-solubilized acetylcholinesterase was broadly distributed in the fractions having Ve/Vo = 1.0-2.0 in gel filtration on a Sepharose 6B column . The intermediary fractions (Ve/Vo = 1.3-1.7) were collected as "the middle active Sepharose 6B eluate" and characterized on the basis of enzymology and protein chemistry . When this eluate was treated with PI-specific phospholipase C, the major activity peak was obtained in the later fractions with Ve/Vo = 1.75-2.0 on the same column chromatography . Lubrol-solubilized and phospholipase C-treated acetylcholinesterase preparations were different in the thermostability, the elution profiles of chromatography on Mono Q, butyl-Toyopearl and phenyl-Sepharose columns, and the affinity to phospholipid micelles . On treatment with PI-specific phospholipase C, Lubrol-solubilized acetylcholinesterase became more thermostable . The phospholipase C-treated enzyme was eluted at lower NaCl concentration from the Mono Q column than the Lubrol-solubilized enzyme . The most important difference was observed in the hydrophobicity of these two enzyme preparations . The Lubrol-solubilized enzyme shows high affinity to phospholipid micelles and hydrophobic adsorbents such as butyl-Toyopearl and phenyl-Sepharose . However, this hydrophobicity was lost when acetylcholinesterase was solubilized from bovine erythrocyte membrane by PI-specific phospholipase C . The presence of myo-inositol was confirmed in the purified preparation of acetylcholinesterase by gas chromatography (GC)-mass spectrometry (MS).(ABSTRACT TRUNCATED AT 250 WORDS)

Jikken Dobutsu, 1987 Oct, 36(4), 387 - 93
Naturally occurring CAR bacillus infection in a laboratory rat colony and epizootiological observations; Itoh T et al.; An epizootic of chronic respiratory disease was found in a rat colony . Lungs of the symptomatic rats showed histopathologically severe peribronchial lymphoid cuffing . Filamentous bacteria were detected on the border of the tracheal and bronchial epithelium by light and electron microscopy . These bacteria did not grown on artificial media but propagated in embryonated chicken eggs . The disease was thus diagnosed as cilia-associated respiratory (CAR) bacillus infection . Epizootiological observations of the natural and experimentally induced cases revealed that the disease was highly contagious, slowly progressive and intractable . Contact infection may play a major role in the transmission of this disease.

J Gen Microbiol, 1987 Oct, 133 ( Pt 10), 2921 - 31
Cloning and expression in Escherichia coli of an insecticidal crystal protein gene from Bacillus thuringiensis var . aizawai HD-133; Chak KF et al.; Using a gene probe derived from the cloned var . sotto insecticidal crystal protein (ICP) gene, we have cloned a Bacillus thuringiensis var . aizawai HD-133 ICP gene in Escherichia coli . The gene encodes a polypeptide that is toxic to Lepidoptera in vivo and in vitro . The protein is expressed at a level sufficient to produce phase-bright inclusions in recombinant E . coli strains, and these inclusions can be partially purified using discontinuous sucrose density gradients . Immunoblotting shows that the inclusions contain a 135 kDa polypeptide which reacts strongly with antiserum raised against the B . thuringiensis var . kurstaki HD-1 P1 polypeptide.

Lab Anim, 1987 Oct, 21(4), 356 - 9
Serodiagnosis of cilia-associated respiratory bacillus infection by the indirect immunofluorescence assay technique; Matsushita S et al.; Antibody to cilia-associated respiratory (CAR) bacillus was detected by the indirect immunofluorescence assay (IFA) technique using tracheal sections of infected mice as antigen in serum samples collected from rats infected naturally and experimentally . Nine of 23 cases of natural infection were positive in IFA antibody, with titres ranging from 1:10 to 1:80, and all these antibody-positive cases were also histologically positive . The remaining 14 cases were negative in both IFA antibody and histological diagnosis, even though some of them were infected with Sendai virus and Mycoplasma pulmonis . In the experimental infection, serum samples collected from 18 rats on days 4, 7, 14, 21, 28 and 56 post-inoculation (PI) (three rats for each point) and examined for IFA antibody revealed that seroconversion occurred in one rat on day 14 PI and in three rats on day 21 PI . Antibody titres of 1:80 to 1:160 remained to the termination of the experiment . The IFA technique was useful for the diagnosis of CAR bacillus infection except in the early stage of the infection.

Eur J Biochem, 1987 Oct 1, 168(1), 153 - 9
Phenylalanine dehydrogenase of Bacillus badius . Purification, characterization and gene cloning; Asano Y et al.; Phenylalanine dehydrogenase produced by Bacillus badius IAM 11059 was purified from the crude extract of B . badius to homogeneity, as judged by disc gel electrophoresis . The enzyme has an isoelectric point of 3.5 and a relative molecular mass, Mr, of 310,000-360,000 . The enzyme is composed of identical subunits with an Mr 41,000-42,000 . The substrate specificity of the enzyme in the oxidative deamination reaction was high for L-phenylalanine, but rather low in the reductive amination reaction, with phenylpyruvate, p-hydroxyphenylpyruvate, and 2-oxohexanoate . The gene for the enzyme was cloned into Escherichia coli with plasmid pBR322 as a vector . The enzyme was expressed in high level in E . coli . The enzyme produced by E . coli transformant was purified to homogeneity and shown to be identical to that of B . badius IAM 11,059 with respect to the specific activity, Mr, subunit structure and amino acid composition.

Surgery, 1987 Oct, 102(4), 622 - 7
Recurrence patterns in a prospective study of patients with stage II breast cancer treated with endocrine-chemotherapy; Crowe JP Jr et al.; Local-regional versus distant recurrence patterns were investigated for 311 patients with stage II node-positive breast cancer who were part of an endocrine-chemotherapy adjuvant breast cancer trial . After mastectomy patients were randomized to receive either cytoxan, methotrexate, and 5-fluorouracil (CMF) (1 year) or CMF with tamoxifen (1 year) with or without bacillus Calmette-Guerin (BCG) . With a median follow-up of 92.1 months, 55.3% of the patients had recurrences . The first site of recurrence was local-regional for 31.4% of patients and distant for 68.6% . This pattern of first recurrence was not associated with treatment groups, menopausal status, race, estrogen receptor value, number of positive lymph nodes, or tumour diameter . Although patients with a first local-regional recurrence had a better overall prognosis compared with those with a first distant recurrence, 52.2% of those patients with an initial local-regional recurrence developed a distant recurrence within 12 months . Among patients who had a recurrence, 48.3% had a local-regional recurrence at some time during their follow-up . Conclusions from this study are (1) patterns of recurrence were not affected by the addition of antiestrogen therapy to chemotherapy; (2) for the variables tested, including number of positive nodes and tumor diameter, no association with recurrence patterns was found; and (3) most patients (52.2%) with a first local-regional recurrence will develop a distant recurrence within 1 year.

Microbiol Sci, 1987 Oct, 4(10), 292 - 5
The mycoplasma genome: Part 2; Christiansen C; This second part of a review of the mycoplasma genome describes protein synthesis, known genes for proteins and the phylogeny of mycoplasmas . The investigation of protein synthesis has led to the very surprising observation that the opal codon is used to code for tryptophan in mycoplasmas . Phylogenetically, the mycoplasmas have been found to descend from Bacillus and related G-positive rods.

Rev Argent Microbiol, 1987 Oct-Dec, 19(4), 148 - 52
{Incidence of Bacillus cereus in powdered dehydrated food}; Iacona VA et al.; Bacillus cereus incidence on dehydrated powdered foods on sale in supermarkets of Santa Fe city was studied . Two hundred and fifty samples of five different foods: desserts, soups, mousses, pre-cooked "polenta" and mashed potatoes, were examined . Toxinogenic activity of strains confirmed as B . cereus by means of the test of lethality in rats, was analyzed . The ratio between contaminated samples and total analyzed samples was always greater than 6% (Table 1) . Besides, none of the analyzed foods exceeded acceptability maximum limit (10(5) UFC/g), established by I.C.M.S.F . (Table 2) . It was checked in all cases that no simple lineal correlation existed between B . cereus and total aerobic bacteria enumerations . On the other hand, the percentage of strains with lethal effects was in all cases greater than 42.8% (Table 3).

Mol Gen Mikrobiol Virusol, 1987 Oct, (10), 27 - 30
{Plasmid transformation of Bacillus cereus protoplasts}; Kovtunenko LV et al.; The process of polyethyleneglycol-induced plasmid transformation of Bacillus cereus protoplasts was studied . Plasmid transfer into Bacillus cereus strains was demonstrated with the frequencies 1.3.10(1)-1.6.10(2) transformants per 1 mkg of plasmid DNA . The plasmids transferred are stably inherited by Bacillus cereus cells causing tetracycline resistance (pBC16) or kanamycin resistance (pUB110 and pBD64) . The proposed method can be used for construction of Bacillus cereus strains having the plasmid determined characteristics.

Lipids, 1987 Oct, 22(10), 698 - 703
Lipid composition of Bacillus megaterium spores and spore membranes; Nikolopoulou M et al.; Bacillus megaterium QM B1551 spore lipids were extracted by an improved technique, and the phospholipid and fatty acid compositions were determined . Phospholipids accounted for 65% of the total fatty acids; the neutral lipid fraction contained 15% and the remaining fatty acids were in the interphase, aqueous phase and pellet from the lipid extraction . Each phospholipid had similar fatty acid compositions as did the delipidated pellet . However, the aqueous phase and, to some extent, the interphase had unique fatty acid compositions . Also, fatty acids were found acylated to proteins, which was observed by electrophoresis of delipidated proteins from spores grown in {1-14C}palmitate . Therefore, spores contain unique non-phosphatide fatty acid components that can now be analyzed.

Cell Struct Funct, 1987 Oct, 12(5), 453 - 61
Light-independent and dependent phases of proplastid development in Euglena gracilis W3BUL; Osafune T et al.; Cells of Euglena gracilis Klebs var . bacillaris Cori mutant W3BUL grown in darkness on Hutner's pH 3.5 medium without agitation accumulate wax ester . These cells have undeveloped proplastid remnants characteristic of this mutant . If these cells are transferred to an inorganic medium and bubbled with 2-3% CO2 in air, the wax disappears and the proplastid expands and develops in darkness to form prolamellar bodies and membrane vessicles within 96 h . No further development takes place in darkness, but if these cultures are illuminated at 96 h formation of prothylakoids is observed . Thus the wax ester accumulated during growth can be used subsequently to support proplastid development up to the prolamellar body stage, but the formation of prothylakoids is strictly light-dependent . Development in this system takes place at a slower rate than in cells grown with shaking and lacking wax which are transferred to resting medium . As previously shown, all of proplastid development requires light under these conditions . It is suggested that the oxygen-requiring utilization of wax in darkness can provide energy and metabolites for a part of proplastid development but the later steps in these cells, or the entire development in cells lacking wax is supported by paramylum degradation which is strictly light-dependent . However, a specific light reaction required for prothylakoid organization is not ruled out.

J Urol, 1987 Oct, 138(4), 867 - 70
Monocyte cytolytic factor in promoting monocyte-mediated lysis of bladder cancer cells by bacillus Calmette-Guerin; Nakamura K et al.; The role of monocytes in cell-mediated cytolysis of bladder cancer cells was investigated . Human peripheral monocytes released a cytolytic factor which lysed T24 bladder cancer cells and a number of human tumor cells, but not normal lymphocytes or fibroblasts . After incubation of monocytes with bacillus Calmette-Guerin for 48 hr . in vitro, cytolysis of T24 cells was increased up to 56.7 +/- 4.1% . Treatment of monocytes with actinomycin D (an inhibitor of RNA transcription) reduced release of cytolytic factor from 27.3 +/- 5.7% cytolysis to 4.5 +/- 1.4% (p less than 0.05) . The response to mitomycin C was different between lymphokines and monocyte cytolytic factor . The mouse monoclonal antibody against human recombinant tumor necrosis factor did not neutralize monocyte cytolytic factor . These results show that this monocyte cytolytic factor is distinct from lymphokines and tumor necrosis factor . The evidence that bacillus Calmette-Guerin increases release of monocyte cytolytic factor may be associated with anti-tumor activity of bacillus Calmette-Guerin in intravesical therapy for treatment of bladder cancer.

J Bacteriol, 1987 Oct, 169(10), 4692 - 5
Role of glutamate dehydrogenase in ammonia assimilation in nitrogen-fixing Bacillus macerans; Kanamori K et al.; Pathways of ammonia assimilation into glutamic acid in Bacillus macerans were investigated by measurements of the specific activities of glutamate dehydrogenase (GDH), glutamine synthetase, and glutamate synthase . In ammonia-rich medium, GDH was the predominant pathway of ammonia assimilation . In nitrogen-fixing cells in which the intracellular NH4+ concentration was 1.4 +/- 0.5 mM, the activity of GDH with a Km of 2.2 mM for NH4+ was found to be severalfold higher than that of glutamate synthase . The result suggests that GDH plays a significant role in the assimilation of NH4+ in N2-fixing B . macerans.

Mol Gen Mikrobiol Virusol, 1987 Oct, (10), 23 - 7
{Homologous and heterologous transcription of the Cry+-plasmid in Bacillus thuringiensis}; Smirnova TA et al.; The possibility of homologous and heterologous transception of Cry+ plasmids in Bacillus thuringiensis is demonstrated . Cry+ plasmids from crystal bearing strain of Bacillus thuringiensis were transferred into acrystalline strain belonging to H5 serotype by mutual incubation . The donor strain was previously marked by the transmissive plasmid pAM beta 1 coding for erythromycin and lincomycin resistance . The transcipients having acquired the ability to synthesize delta-endotoxin were referred to H5 serotype due to their phenotype . By analogous method Cry+ plasmid was transferred from Bacillus thuringiensis to Bacillus cereus . Bacillus cereus strain GP7 was used as a recipient strain resistant to tetracycline . The presence of delta-endotoxin in transcipients was confirmed by bioprobes and immunoenzyme assay . To prove the transfer of Cry+ plasmid the plasmid profiles of the parent strains and transcipients have been analyzed . The formation of cellular contacts during mutual incubation of Bacillus thuringiensis and Bacillus cereus strains was demonstrated by electron microscopic study of ultrafine cuts.

Mol Gen Genet, 1987 Oct, 209(3), 575 - 9
Thermostability and superhelicity of plasmid DNA in Bacillus stearothermophilus; Soutschek-Bauer E et al.; The thermostability of the staphylococcal plasmids pC194 and pUB110 and their antibiotic-resistance determinants was examined upon transfer to Bacillus stearothermophilus CU21 . Plasmid pGS13, a pUB110 derivative carrying the chloramphenicol acetyltransferase (CAT) gene of pC194, could be maintained up to the maximum growth temperature (68 degrees C) by selection for chloramphenicol resistance . In the absence of selective pressure, pGS13 was lost at temperatures above 60 degrees C . Segregational instability of pGS13 was accompanied by a progressive loss of negative superhelicity at elevated temperatures . Thermostable mutants of pGS13 were isolated by screening for expression of the antibiotic-resistance determinants after growth under non-selective conditions . These mutants were found to contain an insertion of a 1.7 kb DNA sequence derived from the cryptic B . stearothermophilus plasmid pBS02 . Increased thermostability correlated with preservation of plasmid superhelicity at elevated temperatures.

Biochem Biophys Res Commun, 1987 Sep 30, 147(3), 1219 - 25
The membrane-anchor of Paramecium temperature-specific surface antigens is a glycosylinositol phospholipid; Capdeville Y et al.; The temperature-specific G surface antigen of Paramecium primaurelia strain 156 was biosynthetically labeled by {3H}myristic acid in its membrane-bound form, but not in its soluble form . It could be cleaved by a phosphatidylinositol-specific phospholipase C from Trypanosoma brucei or from Bacillus cereus which released its soluble form with the unmasking of a particular glycosidic immunodeterminant called the crossreacting determinant . The Paramecium enzyme, capable of converting its membrane-bound form into the soluble one, was inhibited by a sulphydril reagent in the same way as the trypanosomal lipase . From this evidence we propose that the Paramecium temperature-specific surface antigens are anchored in the plasma membrane via a glycophospholipid, and that an endogenous phospholipase C may be involved in the antigenic variation process.

Biochemistry, 1987 Sep 22, 26(19), 6038 - 43
Structure-activity relationships in engineered proteins: characterization of disruptive deletions in the alpha-ammonium group binding site of tyrosyl-tRNA synthetase; Lowe DM et al.; Residues Asp-78 and Gln-173 of the tyrosyl-tRNA synthetase of Bacillus stearothermophilus form part of the binding site for tyrosine by making hydrogen bonds with the alpha-ammonium group . Asp-38 is close enough to the group to make an important electrostatic contribution . Unlike other residues in the active site that have been studied by site-directed mutagenesis, Asp-38, Asp-78, and Gln-173 are part of hydrogen-bonded networks . Each of these residues has been mutated to an alanine, and the resultant mutants have been studied by kinetics to construct the difference energy diagrams for the formation of tyrosyl adenylate . In each example, the binding of tyrosine is weakened by about 2.5 kcal mol-1 . But, unlike previous mutants, the dissociation of the second substrate, in this case ATP, is also seriously affected, being weakened by some 2 kcal mol-1 for TyrTS(Ala-78) and TyrTS(Ala-173) . The energy of the transition state for the formation of tyrosyl adenylate is raised by 7.8 kcal mol-1 for the former and 4.5 kcal mol-1 for the latter mutant . Addition of these mutants to linear free energy plots constructed for the nondisruptive mutants in the accompanying study {Fersht, A . R., Leatherbarrow, R . J., & Wells, T . N . C . (1987) Biochemistry (preceding paper in this issue)} reveals large deviations of the data for TyrTS(Ala-38) and TyrTS(Ala-78) from the regression line . These thus belong to a different class of mutations from previous nondisruptive examples . This observation combined with the structural evidence and difference energy diagrams strongly suggests that the mutations Asp----Ala-38 and Asp----Ala-78 are disruptive in nature.

J Immunol, 1987 Sep 15, 139(6), 2032 - 7
Importance of L3T4+ and Lyt-2+ cells in the immunologic control of infection with Mycobacterium bovis strain bacillus Calmette-Guérin in mice . Assessment by elimination of T cell subsets in vivo; Pedrazzini T et al.; The course of infection after injection of small doses of bacillus Calmette-Guerin (BCG) was studied in mice which were depleted in vivo of T cell subsets by administration of either anti-L3T4 or anti-Lyt-2 mAb . The results presented herein strongly suggest that the L3T4+ subpopulation play a pivotal role in the immunologic control of BCG infection because the depletion of L3T4+ cells led to a dramatic increase in the number of viable bacteria . Depletion of Lyt-2+ cells had no significant effect on the course of infection . These results were confirmed by using adoptive transfer experiments which showed that protective immunity was mediated by L3T4+ cells generated in the spleen as a result of infection . Moreover, T cells capable of controlling the recurrence of BCG multiplication from residual bacteria remaining in organs after the recovery from infection were shown to belong to the L3T4+ subpopulation.

J Immunol, 1987 Sep 15, 139(6), 1885 - 90
Modulation of mouse peritoneal macrophage Ia and human peritoneal macrophage HLA-DR expression by alpha 2-macroglobulin "fast" forms; Hoffman MR et al.; alpha 2-Macroglobulin (alpha 2M) is converted from its native form into electrophoretically "fast" forms by reaction with proteinases or with methylamine . The "fast" forms both bind to specific receptors on macrophages (MP) . We have previously shown that alpha 2M "fast" forms modulate effector functions of murine peritoneal MP . In the present study, alpha 2M "fast" forms antagonized the increase in MP HLA-DR and Ia expression induced in vitro by interferon-gamma (IFN-gamma) . This effect was observed with human peritoneal MP, as well as MP from peptone-injected and bacillus Calmette-Guerin-infected mice of three strains . alpha 2M-trypsin, which had been reacted with aprotinin and alpha 2M-methylamine, both of which lack proteolytic activity, also antagonized interferon-induced Ia expression . alpha 2M "fast" forms also reduced the ability of MP to serve as accessory cells for lectin-induced lymphocyte proliferation . alpha 2M "fast" form is an immune modulator of human and murine MP function, probably through a specific receptor-mediated mechanism.

Biochim Biophys Acta, 1987 Sep 11, 925(3), 356 - 61
Properties of gamma-glutamyl arylamidase activity of the heavy subunit of gamma-glutamyl arylamidase from Bacillus sp . strain No . 12; Hwang SY et al.; gamma-Glutamyl arylamidase of Bacillus sp . strain No . 12, composed of two heavy (Mr 56,000) and two light (Mr 46,000) subunits, was dissociated and inactivated by mild SDS treatment . The activity was restored in the isolated heavy subunit but not in the light subunit when SDS was removed by dialysis . The restored activity of the heavy subunit was similar to that of the native enzyme with regard to substrate specificity and inhibition and activation by alpha- and gamma-glutamyl compounds, free amino acids, peptides, enzyme inhibitors, and anti-native enzyme antibody.

Ann Emerg Med, 1987 Sep, 16(9), 1016 - 22
Bacteriology of the freshwater environment: implications for clinical therapy; Auerbach PS et al.; Water and animal tissue samples were obtained from sources in Tennessee, California, and Florida . Purified bacterial colonies were isolated and organisms identified . Fifty-eight isolates were recovered . Twenty-seven Gram-negative isolates were identified . Gram-positive organisms were of the coryneform group or Bacillus species . Antibiotic susceptibility testing showed that Aeromonas species were relatively resistant to a wide variety of antimicrobials, which included trimethoprim, cefazolin, and ampicillin . Antibiotics effective against more than 90% of Gram-negative isolates included ciprofloxacin, imipenem, ceftazidime, and trimethoprim-sulfamethoxazole . Freshwater Gram-positive organisms did not display any unexpected susceptibility features . Recommendation for treatment are based on sensitivity in culture and the potentially serious nature of infections caused by Aeromonas species.

Cancer, 1987 Sep 1, 60(5), 1009 - 16
The practical use of tumor marker determination in bladder washing specimens . Assessing the urothelium of patients with superficial bladder cancer; Orihuela E et al.; Bladder washing specimens from 81 patients with recurrent multifocal superficial bladder cancer were evaluated for DNA profile by flow cytometry and cell-surface blood group (ABH) antigen reactivity by a modified specific erythrocyte adherence test . The study was conducted in a prospective, blind, nonrandomized fashion . Fifteen patients were treated with transurethral resection (TUR) alone and 66 with TUR and intravesical administration of bacillus Calmette-Guerin weekly for 6 weeks (TUR + BCG) . Among the patients treated with TUR only, there was a notably greater rate of tumor recurrence and progression in patients with unfavorable tumor markers (aneuploidy and ABH-negative reactivity) than in those with favorable markers (diploidy and ABH-positive reactivity) . The difference was less striking in patients treated with BCG, which reduced the frequency of recurrence or progression at 30 months from 87% to 44% and from 60% to 23%, respectively . This favorable effect of BCG was virtually confined to patients with initially favorable markers and to those whose initially unfavorable markers became favorable during BCG administration . Aneuploidy and negative ABH are phenotypic expressions of undifferentiation that can forecast the potential of the urothelium to form new tumors and predict invasion . To an extent, these markers are independent of the grade and stage of the disease . BCG can induce prognostically favorable conversion of the markers expression . Lack of such conversion indicates lack of response to BCG and should be regarded as evidence of persistent disease even if conventional methods do not reveal it . Therefore, sequential determination of markers is useful in monitoring patients with superficial bladder cancer treated with intravesical BCG.

J Am Mosq Control Assoc, 1987 Sep, 3(3), 485 - 8
Efficacy of Arosurf MSF and formulations of Bacillus thuringiensis var . israelensis against Anopheles albimanus: laboratory bioassay; Perich MJ et al.; The efficacy of Arosurf MSF alone and in combination with three preparations of Bacillus thuringiensis var . israelensis (B.t.i.) against Anopheles albimanus larvae, pupae and eggs was determined by bioassay . Arosurf MSF alone was effective against the egg, 4th larval instar and pupal stages . All Arosurf MSF and B.t.i . combined formulations produced over 90% mortality of all larvae and pupae, 48 hr posttreatment . Egg eclosion was reduced to approximately 25% with all formulations containing Arosurf MSF.

Mol Microbiol, 1987 Sep, 1(2), 187 - 94
Identification, cloning and sequence analysis of the Bacillus sphaericus 1593 41.9 kD larvicidal toxin gene; Hindley J et al.; A number of strains of the widespread aerobic soil bacterium, Bacillus sphaericus, possess crystalline inclusions of a toxin lethal to a variety of insect (larvae) which are vectors of major tropical diseases . Partial amino acid sequence data from one strain, B . sphaericus 2362 have permitted us to design oligonucleotide probes for identifying the toxin gene in the closely related B . sphaericus 1593 . The gene was found to be contained within an EcoRI-HindIII fragment and was cloned in its entirety in the bacterial plasmid pUC12 . The DNA sequence was determined together with the upstream and downstream controlling elements, and a sequence of 370 amino acids was deduced for the toxin protein . This is the first reported sequence of a B . sphaericus toxin gene and will facilitate further work in characterizing the genes from other strains of different virulence and host range . The data do not support the suggestion that the toxin is derived by proteolysis of a protoxin precursor.

Mikrobiologiia, 1987 Sep-Oct, 56(5), 816 - 8
{Spontaneous bacteriophage induction in Bacillus thuringiensis}; Besaeva SG et al.; The production of temperate bacteriophages was studied in the process of batch cultivation of three Bacillus thuringiensis lysogenic strains . Phage titres were determined using an indicator culture (IPM-1148) . The growth of bacteriophages was induced when thermoactivated spores germinated . Some cells (1.10(-3)-2.10(-3)) underwent lysis without their division . The subsequent lytic cycles occurred in the actively growing culture . Phage titres ceased to rise before the exponential growth phase was over.

J Appl Bacteriol, 1987 Sep, 63(3), 207 - 15
Relation of the heat resistance of bacterial spores to chemical composition and structure . II . Relation to cortex and structure; Mallidis CG et al.; The relation between the amount of cortex, measured as total hexosamine, as diaminopimelic acid and as muramic lactam, and the heat resistance of spores of five different strains of Bacillus stearothermophilus was studied . Electron micrographs of thin sections of the spores were made to relate the structure of the spores to chemical and thermal characteristics . It was found that the amount of the cortex was significantly related to heat resistance of the spores . Strains with more electron-dense and better organized cortices were found to express higher heat resistance.

Infect Dis Clin North Am, 1987 Sep, 1(3), 575 - 90
Cat scratch disease; Moriarty RA et al.; In summary, CSD is a relatively common cause of localized lymphadenopathy, with 80 per cent of cases occurring in children . This self-limited infection is caused by a small pleomorphic bacillus that has been identified in ocular granuloma, skin, and lymph node specimens . Unusual manifestations of the disease such as the oculoglandular disease of Parinaud, encephalopathy, or severe systemic disease occur in about ten per cent of patients . Management consists of symptomatic treatment and occasionally aspiration of a node that suppurates . The disease usually resolves spontaneously in 2 to 4 months.

Rev Infect Dis, 1987 Sep-Oct, 9(5), 884 - 90
Dysgonic fermenter 2 septicemia; Hicklin H et al.; Dysgonic fermenter 2 (DF-2) is a slow-growing gram-negative bacillus causing a zoonotic infection that is acquired through dog bites or other contact with dogs . Splenectomized patients and those with alcoholic liver disease are most susceptible to DF-2 infection . The clinical picture can be one of fulminant septicemia and disseminated intravascular coagulation in the splenectomized patient; the presentation is milder in the alcoholic patient . The overall mortality from DF-2 septicemia among the 41 cases reported in the literature is 27% . The organism is sensitive to penicillin, resistant to aminoglycosides, and not easily grown on common media . It appears to be serum-sensitive in tests with normal human serum . Penicillin prophylaxis of dog bite wounds is especially important in high-risk patients . DF-2 infection should be considered when any splenectomized patient develops fulminant septicemia, disseminated intravascular coagulation, and peripheral gangrene . Examination of a gram stain of the peripheral blood or buffy coat is of value in such cases.

Biokhimiia, 1987 Sep, 52(9), 1454 - 60
{Determination of inhibition sites during hydrolysis of polydeoxyribonucleotides by exonucleases III from Bacillus amyloliquefaciens and Escherichia coli}; Chikaev NA et al.; The influence of the primary structure of polydeoxyribonucleotides on the rate of hydrolysis with exonuclease III from Bacillus amyloliquefaciens and Escherichia coli was investigated . The substrates used were synthetic oligodeoxyribonucleotides and pBR 322 DNA fragments labeled with 32P at the 5'-termini of one of the chains . According to the data from polyacrylamide gel electrophoresis performed under denaturing conditions, the hydrolysis of these substrates by unsaturating concentrations of B . amyloliquefaciens and E . coli exonuclease III proceeds with several reproducible "stops" . The decrease of the reaction rate was shown to take place just before the pyrimidine blocks in the digested DNA chain.

Arch Intern Med, 1987 Sep, 147(9), 1642 - 4
Gentamicin resistance among gram-negative bacillary blood isolates in a hospital with long-term use of gentamicin; Mylotte JM; Between 1977 and 1985, gentamicin was the only formulary aminoglycoside at the Buffalo Veterans Administration Medical Center . During this time, there was a significant increase in the amount of gentamicin purchased . Amikacin represented 11% or less of the total aminoglycoside purchased in the same period, but purchases of this agent also significantly increased . Because of this long-term use of gentamicin, a retrospective analysis of gentamicin resistance among gram-negative bacillary blood isolates was performed . The results of this review revealed no significant change in the overall incidence of gram-negative bacteremia; approximately 75% of these bacteremic episodes were hospital acquired . The mean yearly gentamicin-resistance rate of gram-negative blood isolates was 13.2% (range, 6% to 18%) with no significant change in the rate for the period reviewed . However, for certain strains there were fluctuations in the percentage of resistance from year to year, suggesting that clusters of infections due to these organisms had occurred . Bacteremic infection due to resistant organisms was a major contributor to the overall level of gentamicin resistance among blood isolates . Amikacin resistance among gram-negative blood isolates was rare . In conclusion, despite the predominant use of gentamicin there was no change in the gentamicin resistance rate among gram-negative bacillary blood isolates during a nine-year period . The rate of gentamicin resistance among blood isolates appeared to be related to outbreaks/clusters of infections due to resistant strains rather than the frequency of use of gentamicin.

J Infect Dis, 1987 Sep, 156(3), 456 - 62
Antibiotic therapy, endotoxin concentration in cerebrospinal fluid, and brain edema in experimental Escherichia coli meningitis in rabbits; Tauber MG et al.; We investigated the effect of cefotaxime and chloramphenicol on endotoxin concentrations in cerebrospinal fluid (CSF) and on the development of brain edema in rabbits with Escherichia coli meningitis . Both antibiotics were similarly effective in reducing bacterial titers . Cefotaxime, but not chloramphenicol, induced a marked increase of endotoxin in CSF, from log10 1.5 +/- 0.8 to log10 2.8 +/- 0.7 ng/ml (P less than .01) . This result was associated with an increase in brain water content (405 +/- 12 g of water/100 g of dry weight compared with 389 +/- 8 g in untreated controls; P less than .01), whereas in animals treated with chloramphenicol, brain water content was identical to controls . The cefotaxime-induced increase in endotoxin concentration and brain edema were both neutralized by polymyxin B, which binds to the lipid A moiety of endotoxin, or by a monoclonal antibody to lipid A . These results indicate that treating gram-negative bacillary meningitis with selected antibiotics induces increased endotoxin concentrations in CSF that are associated with brain edema.

Microbiol Sci, 1987 Sep, 4(9), 274 - 6
Bacillus larval toxin crystal protein; Klier A et al.; During sporulation some bacteria produce parasporal inclusions which are toxic for insect larvae . The proteins responsible for this toxicity have been characterized and their genes have been cloned . Using genetic engineering methods, it is now possible to establish new bacterial strains and to introduce these genes into plant genomes.

Mol Pharmacol, 1987 Sep, 32(3), 437 - 42
Effect of auranofin and other gold complexes on the activity of phospholipase C; Snyder RM et al.; Auranofin (AF) is an orally active chrysotherapeutic agent used for the treatment of rheumatoid arthritis, a self-perpetuating inflammatory disease . Because of reports suggesting that AF and other gold complexes can, under certain circumstances, exacerbate rheumatoid inflammatory lesions in humans and adjuvant arthritic rats and that phospholipase C (PLC) and phospholipase A2 activities are increased in rheumatoid patients, the effects of AF and a related gold complex on in situ mammalian and purified Bacillus cereus PLC were examined . Results of our studies show that 1) AF and triethylphosphine gold chloride (TEPG), an AF analog, stimulated PLC activity in the sonicate of RAW 264.7 macrophages; 2) AF and TEPG stimulated B . cereus PLC activity in a concentration-dependent manner, but the pattern of stimulation and concentrations of drugs required to stimulate the purified enzyme differ from those seen with the macrophage PLC; 3) metals (cobalt and zinc) and sulfhydryl reagents (N-ethylmaleimide, iodoacetic acid, and glutathione), tested at the same concentrations of AF that enhanced PLC activity, had no effect on the enzyme . These data suggest that stimulation of PLC may be a generic phenomenon since two divergent PLCs are affected by gold complexes . Additionally, these studies may provide one potential explanation for rheumatoid lesion flares seen in patients and animals on chrysotherapy.

Am J Med, 1987 Sep, 83(3), 499 - 502
Appropriateness of antibiotic therapy in long-term care facilities; Jones SR et al.; The objective of this study was to examine the appropriateness of antibiotic therapy in nursing homes . Information was abstracted from infection control reports and patients' charts for a three-month period at two nursing homes in Portland, Oregon . A panel of two board-certified infectious disease specialists and one hospital pharmacist reviewed the information and rated the appropriateness of each prescription using a previously developed scale . Among the 120 infections, treatment was rated as appropriate in 49 percent, as inappropriate in 42 percent, and as unjustified in 9 percent . Cephalosporins received the lowest percent of appropriate ratings (27 percent) . The primary reason for an inappropriate rating was that a more effective drug was recommended for empiric therapy of gram-negative bacillary infections . These bacteria are often resistant to oral antibiotics . There were no significant differences in appropriateness by type of organism, infection site, or clinical outcome . Physician education and the development of systems and guidelines for optimal management in this population are appropriate actions for the future.

J Bacteriol, 1987 Sep, 169(9), 4110 - 8
Expression of a cloned Bacillus thuringiensis crystal protein gene in Escherichia coli; Schnepf HE et al.; The expression in Escherichia coli of a cloned crystal protein gene from Bacillus thuringiensis was investigated through the use of fusions of the crystal protein gene promoter to beta-galactosidase and catechol oxidase genes . Analysis of deletion and insertion derivatives of the crystal protein gene promoter showed that a region of B . thuringiensis DNA located between 87 and 258 base pairs upstream from the transcription initiation site caused reduced transcription from this promoter . Insertion of Tn5 145 base pairs upstream from the transcription initiation site resulted in overproduction of the crystal protein . S1 nuclease mapping experiments failed to detect transcription from an outwardly directed promoter in Tn5, indicating that the overproduction resulted from the disruption or repositioning of the transcription-suppressing region.

Int J Lepr Other Mycobact Dis, 1987 Sep, 55(3), 481 - 93
Type 1 reactions in leprosy--heterogeneity in T-cell functions related to the background leprosy type; Laal S et al.; Nineteen each of paucibacillary borderline tuberculoid (BT) and multibacillary borderline borderline (BB)/borderline lepromatous (BL) leprosy patients undergoing type 1 reactions were compared with nonreactional stable patients of the appropriate leprosy type . In the BT reactional group, both phytohemagglutinin-induced and, more importantly, antigen-induced lymphoproliferation was reduced in 80%-90% of the patients . On the other hand, leukocyte migration inhibition was reduced in 40% and remained unchanged in the others . Suppressor-cell activity as evaluated by a costimulant assay was also reduced in a majority of the reactional BT individuals . In contrast, the bacilliferous BB and BL patients in reaction showed significant general improvement in leukocyte migration inhibition (p less than 0.001) and antigen-induced lymphoproliferation (p less than 0.05) as compared to the expected hyporesponsive/anergic uncomplicated BB-BL patients . Suppressor-cell activity also recovered during the reactional phase . However, no significant differences were observed in either of the reactional or stable leprosy types in the numbers of total T cells (OKT3+) and their subsets as defined by OKT4+ (helper/inducer) and OKT8+ (suppressor/cytotoxic) functional phenotypes . Moreover, during type 1 reactions the 48-hr delayed-type hypersensitivity (DTH) responses after intradermal injection of Mycobacterium leprae antigens continued to reflect the background leprosy type rather than the functional perturbations in the circulating T cells . Only a marginal increase in DTH was observed in some BT reactional individuals . No consistent pattern in the above in vitro T-cell-related responses was discernable in the same individuals 4-6 months after subsidence of reactions . The clinical entity of type 1 reactions encompassing paucibacillary and multibacillary leprosy shows a heterogeneity/dichotomy in T-cell responses which may reflect different immunological mechanisms underlying the reactional state.

J Bacteriol, 1987 Sep, 169(9), 4399 - 402
Nucleotide sequence of the beta-cyclodextrin glucanotransferase gene of alkalophilic Bacillus sp . strain 1011 and similarity of its amino acid sequence to those of alpha-amylases; Kimura K et al.; The nucleotide sequence of the gene for cyclodextrin glucanotransferase of alkalophilic Bacillus sp . strain 1011 was determined . The deduced amino acid sequence at the NH2-terminal side of the enzyme showed a high homology with the sequences of alpha-amylase in the three regions which constitutes the active centers of alpha-amylases.

J Bacteriol, 1987 Sep, 169(9), 4342 - 8
High-affinity potassium uptake system in Bacillus acidocaldarius showing immunological cross-reactivity with the Kdp system from Escherichia coli; Bakker EP et al.; During growth with low levels of K+, Bacillus acidocaldarius expressed a high-affinity K+ uptake system . The following observations indicate that this system strongly resembles the Kdp-ATPase of Escherichia coli: (i) its high affinity for K+ (Km of 20 microM or below); (ii) its poor transport of Rb+; (iii) the enhanced ATPase activity of membranes derived from cells grown with low levels of K+ (this activity was stimulated by K+ and inhibited by vanadate); (iv) the expression of an extra protein with a molecular weight of 70,000 in cells grown with low levels of K+; and (v) the immunological cross-reactivity of this 70,000-molecular-weight protein with antibodies against the catalytic subunit B of the E . coli Kdp system . Antibodies against the complete E . coli Kdp system, which immunoprecipitated the whole E . coli KdpABC complex, almost exclusively precipitated the 70,000-molecular-weight protein from detergent-solubilized B . acidocaldarius membranes . The possibility that the B . acidocaldarius Kdp system consists of a single, KdpB-type subunit is discussed.

J Am Mosq Control Assoc, 1987 Sep, 3(3), 407 - 11
Dose-mortality responses of crawfish and mosquitoes to selected pesticides; Holck AR et al.; A study was conducted to determine the toxicities (LC50S) of several pesticides on the commercially important red swamp crawfish, Procambarus clarkii, and 3 mosquito species common in Louisiana ricelands--Anopheles quadrimaculatus, Culex salinarius and Psorophora columbiae . Pesticides tested in laboratory bioassays included Bacillus sphaericus, B . thuringiensis var . israelensis, bendiocarb, glyphosate, isostearyl alcohol, malathion, propoxur, resmethrin synergized with piperonyl butoxide (PBO) and thiobencarb . Isostearyl alcohol was the least toxic compound to crawfish, with a LC50 of greater than 10,000 ppm, while resmethrin + PBO (1:3 ratio) was the most toxic with a LC50 of 0.00082 ppm . The herbicides glyphosate and thiobencarb were the least toxic compounds for the mosquito species tested, while B . t . var . israelensis and resmethrin + PBO were the most toxic.

Mol Gen Genet, 1987 Sep, 209(2), 396 - 8
Expression of the larvicidal gene of Bacillus sphaericus 1593M in the cyanobacterium Anacystis nidulans R2; Tandeau de Marsac N et al.; A 3.6 kb HindIII DNA fragment from Bacillus sphaericus 1593M was cloned and expressed in Escherichia coli and B . subtilis using pHV33 as shuttle vector and in the cyanobacterium Anacystis nidulans R2 with pUC303 as shuttle vector . The level of toxin activity of the respective recombinant plasmids pGsp04 and pGsp12 against Culex mosquito larvae was found to be the same in Escherichia coli and in the cyanobacterium.

Arch Biochem Biophys, 1987 Sep, 257(2), 357 - 62
Enzymatic conversion of the antibiotic metronidazole to an analog of thiamine; Alston TA et al.; We propose that adverse effects of the antibiotic metronidazole may be due, wholly or in part, to its conversion to a thiamine analog and consequent vitamin B1 antagonism . Consistent with this hypothesis, the drug is accepted as a substrate for the thiaminase (EC 2.5.1.2) elaborated as an exoenzyme by the human gut flora constituent Bacillus thiaminolyticus and is also a substrate for the intracellular thiaminase of the mollusk Venus mercenaria . The product, identified as the 1-{(4-amino-2-methyl-5-pyrimidinyl)methyl}-3-(2-hydroxyethyl)-2-methyl-4 - nitroimidazolium cation, is a close structural analog of thiamine and is an effective inhibitor of thiamine pyrophosphokinase in vitro . Due to its susceptibility to nucleophilic attack, the analog is unstable, releasing inorganic nitrite under mild conditions . Enzymatic alkylation reactions such as that effected by thiaminase may have general pharmacological significance as a route of increasing the electrophilicity and/or reduction potential of drugs which are heterocyclic weak bases.

Biochimie, 1987 Sep, 69(9), 1001 - 6
Three-dimensional image reconstruction from ordered arrays of 70S ribosomes; Arad T et al.; A better understanding of the molecular mechanism of protein biosynthesis still awaits a reliable model for the ribosomal particle . We describe here the application of a diffraction technique, namely three-dimensional image reconstruction from two-dimensional sheets of 70S ribosomes from Bacillus stearothermophilus at 47 A resolution . The three-dimensional model obtained by these studies shows clearly the two subunits, the contact points between them, an empty space large enough to accommodate the components of protein biosynthesis, the location of regions rich in RNA and a possible binding site for mRNA . The tunnel within the 50S particle which may provide the path taken by the nascent polypeptide chain in partially resolved.

J Bacteriol, 1987 Sep, 169(9), 4092 - 8
Molecular sieving through S layers of Bacillus stearothermophilus strains; Sara M et al.; The permeability properties and the exclusion limits of the crystalline surface layers (S layers) of two selected strains of Bacillus stearothermophilus were investigated . Measurements were performed of passive solute uptake into the intracellular space of native or glutaraldehyde-treated sacculi . Native sacculi were prepared from whole cells by extracting the cytoplasmic membrane with Triton X-100 under conditions which preserved the integrity of the S layer and the peptidoglycan-containing layer . The permeability barrier was found to consist of three adjacent layers, namely, the S layer, the peptidoglycan-containing layer, and an incomplete S layer attached to the inner face of the peptidoglycan-containing layer . In glutaraldehyde-treated sacculi the peptidoglycan was digested after stabilizing the S-layer lattice by chemical cross-linking . The solutes selected for the uptake measurements were mannose, proteins, and dextrans of increasing molecular weights . The S layers of both strains allowed free passage for molecules with a molecular weight of up to 30,000 and showed sharp exclusion limits between molecular weights of 30,000 and 45,000, suggesting a limiting pore diameter of about 4.5 nm.

J Bacteriol, 1987 Sep, 169(9), 4061 - 7
Cloning of the gene for the larvicidal toxin of Bacillus sphaericus 2362: evidence for a family of related sequences; Baumann P et al.; During sporulation, Bacillus sphaericus 2362 produces a parasporal crystalline protein which is toxic for the larvae of a number of mosquito species . Using the Escherichia coli cloning vector lambda gt11, in which gene products of the inserts may be fused to beta-galactosidase, we isolated 29 bacteriophages which produced peptides-reacting with antiserum to crystal protein . On the basis of restriction enzyme analyses of the recombinants and Ouchterlony immunodiffusion experiments with induced lysogens as a source of antigens, the recombinants were assigned to three groups, designated A, B, and C . Group A consisted of three clones which appeared to express all or part of the B . sphaericus toxin gene from their own promoters and one clone producing a beta-galactosidase-toxin fusion protein . The host cells of two induced recombinant lysogens of this group were toxic to larvae of Culex pipiens . A cell suspension containing 174 ng (dry weight) of the more toxic recombinant per ml killed 50% of the larvae . Both recombinants formed peptides with molecular sizes of 27, 43, and 63 kilodaltons (kDa) . The antigenically related 27- and 43-kDa peptides were distinct from the 63-kDa peptide, which resembled crystals from sporulating cells of B . sphaericus in which antigenically distinct 43- and 63-kDa proteins are derived from a 125-kDa precursor . A 3.5-kilobase HindIII fragment from recombinants having toxic activity against larvae was subcloned into pGEM-3-blue . E . coli cells harboring this fragment were toxic to mosquito larvae and produced peptides of 27, 43, and 63 kDa . The distribution of the A gene among strains of B . sphaericus of different toxicities suggested that it is the sole or principal gene encoding the larvicidal crystal protein . The two recombinants of group B and the 23 of group C were all beta-galactosidase fusion proteins, suggesting that in E . coli these genes were not readily expressed from their own promoters . The distribution of these two genes in different strains of B . sphaericus suggested that they do not have a role in the toxicity of this species to mosquito larvae.

FEBS Lett, 1987 Aug 31, 221(1), 179 - 83
Repressor gene, blaI, for Bacillus licheniformis 749 beta-lactamase; Nicholls NJ et al.; The repressor gene, blaI, for the beta-lactamase of Bacillus licheniformis 749 was functional when cloned in Escherichia coli, but addition of a beta-lactam did not lead to induction . One plasmid contained fragments from the inducible strain (source of repressor), the other carried fragments from the blaI- mutant 749/C (target) . blaI lies just 5' to the promoter for the structural gene, blaP, and the target is the promoter region between the two genes . Interaction with both promoters seemed necessary for full repression . BlaI is a hydrophilic protein (Mr 15036) with the some structural similarities to repressors from Gram-negative bacteria.

Biochim Biophys Acta, 1987 Aug 21, 914(3), 294 - 8
A strong carboxylate-arginine interaction is important in substrate orientation and recognition in lactate dehydrogenase; Hart KW et al.; Using site-directed mutagenesis, Arginine-171 at the substrate-binding site of Bacillus stearothermophilus, lactate dehydrogenase has been replaced by lysine . In the closely homologous eukaryotic lactate dehydrogenase, this residue binds the carboxylate group of the substrate by forming a planar bifurcated bond . The mutation diminishes the binding energy of pyruvate, alpha-ketobutyrate and alpha-ketovalerate (measured by kcat/Km) by the same amount (about 6 kcal/mol) . For each additional methylene group on the substrate, there is a loss of about 1.5 kcal/mol of binding energy in both mutant and wild-type enzymes . From these parallel trends in the two forms of enzyme, we infer that the mode of productive substrate binding is identical in each, the only difference being the loss of a strong carboxylate-guanidinium interaction in the mutant . In contrast to this simple pattern in kcat/Km, the Km alone increases with substrate-size in the wild-type enzyme, but decreases in the mutant . These results can be most simply explained by the occurrence of relatively tight unproductive enzyme-substrate complexes in the mutant enzyme as the substrate alkyl chain is extended . This does not occur in the wild-type enzyme, because the strong orienting effect of Arg-171 maximizes the frequency of substrates binding in the correct alignment.

Eur J Biochem, 1987 Aug 17, 167(1), 123 - 4
Crystallization of and X-ray investigations on glucose dehydrogenase from Bacillus megaterium; Pal GP et al.; The tetrameric glucose dehydrogenase from Bacillus megaterium M1286 belongs to the 'short' family of dehydrogenases with 262 amino acids per subunit (Mr approximately 30,000), and does not require Zn2+ for enzymatic action . It was crystallized as complex with its coenzyme NAD from a 1-2% protein solution by the batch method using ammonium sulfate as precipitant at pH 6.5 . Crystals appeared within two days as clusters of large plates with maximum dimensions of 2 mm, which diffract X-rays to a resolution of at least 0.2 nm . The space group is orthorhombic P2(1)2(1)2(1), unit-cell dimensions are a = 15.03 nm, b = 10.42 nm and c = 6.74 nm . Assuming one molecule (approximately 120 kDa) per asymmetric unit the VM value is 0.0022 nm3/Da and the solvent content of the crystals is 45% based on a partial specific volume for the protein of 0.723 ml/g . The crystallization was further improved by using the microdialysis technique where instead of clusters, single crystals appeared within 7 days.

J Am Vet Med Assoc, 1987 Aug 15, 191(4), 431 - 4
Bacillus piliformis infection (Tyzzer's disease) in a calf; Webb DM et al.; A 1-week-old Jersey bull calf with a history of diarrhea, weakness, and lethargy was submitted for necropsy . Principal macroscopic findings were enteritis and multifocal necrotizing hepatitis . Histologically and ultrastructurally, organisms with characteristics of Bacillus piliformis were associated with the foci of necrosis in the liver.

J Biol Chem, 1987 Aug 15, 262(23), 11038 - 45
Ammonia assimilation in Bacillus polymyxa . 15N NMR and enzymatic studies; Kanamori K et al.; Pathways of ammonia assimilation into glutamic acid and alanine in Bacillus polymyxa were investigated by 15N NMR spectroscopy in combination with measurements of the specific activities of glutamate dehydrogenase, glutamine synthetase, glutamate synthetase, alanine dehydrogenase, and glutamic-alanine transaminase . Ammonia was found to be assimilated into glutamic acid predominantly by NADPH-dependent glutamate dehydrogenase with a Km of 2.9 mM for NH4+ not only in ammonia-grown cells but also in nitrate-grown and nitrogen-fixing cells in which the intracellular NH4+ concentrations were 11.2, 1.04, and 1.5 mM, respectively . In ammonia-grown cells, the specific activity of alanine dehydrogenase was higher than that of glutamic-alanine transaminase, but the glutamate dehydrogenase/glutamic-alanine transaminase pathway was found to be the major pathway of 15NH4+ assimilation into {15N}alanine . The in vitro specific activities of glutamate dehydrogenase and glutamine synthetase, which represent the rates of synthesis of glutamic acid and glutamine, respectively, in the presence of enzyme-saturating concentrations of substrates and coenzymes are compared with the in vivo rates of biosynthesis of {15N}glutamic acid and {alpha,gamma-15N}glutamine observed by NMR, and implications of the results for factors limiting the rates of their biosynthesis in ammonia- and nitrate-grown cells are discussed.

Biochem J, 1987 Aug 15, 246(1), 83 - 8
The inhibition of glucokinase and glycerokinase from Bacillus stearothermophilus by the triazine dye Procion Blue MX-3G; Goward CR et al.; Glucokinase from Bacillus stearothermophilus was irreversibly inactivated by the reactive dichlorotriazinyl dye Procion Blue MX-3G at pH 8.0 . The enzyme was protected from inactivation by the substrate MgATP . Kinetic data implied that the dye occupied the MgATP-binding site . The apparent Km values for MgATP and D-glucose were found to be 70 microM and 210 microM respectively, and the Kd of the pure reactive dye was 16 microM; 1 mol of the pure reactive dye bound to 1 mol of glucokinase subunit . The dye was shown to have potential as an affinity probe for glucokinase . Glycerokinase from the same bacterium was inactivated by Procion Blue MX-3G at high concentrations (5 mM), but only after a period of increased enzyme activity . Kinetic data indicated that the dye preferentially attacked the glycerol-binding site . The apparent Km values for MgATP and glycerol were found to be 38 microM and 13 microM respectively, and 4 mol of reactive dye could be bound to 1 mol of glycerokinase subunit . This was surprising in view of the MgATP-dependent elution of glycerokinase from immobilized Procion Blue MX-3G.

Nucleic Acids Res, 1987 Aug 11, 15(15), 6049 - 62
Purification and DNA binding properties of the blaI gene product, repressor for the beta-lactamase gene, blaP, of Bacillus licheniformis; Grossman MJ et al.; The location of the repressor gene, blaI, for the beta-lactamase gene blaP of Bacillus licheniformis 749, on the 5' side of blaP, was confirmed by sequencing the bla region of the constitutive mutant 749/C . An amber stop codon, likely to result in a nonfunctional truncated repressor, was found at codon 32 of the 128 codon blaI open reading frame (ORF) located 5' to blaP . In order to study the DNA binding activity of the repressor, the structural gene for blaI, from strain 749, with its ribosome binding site was expressed using a two plasmid T7 RNA polymerase/promotor system (S . Tabor and C . C . Richardson . Proc . Natl . Acad . Sci . 82, 1074-1078 (1985) . Heat induction of this system in Escherichia coli K38 resulted in the production of BlaI as 5-10% of the soluble cell protein . Repressor protein was then purified by ammonium sulfate fractionation and cation exchange chromatography . The sequence of the N-terminal 28 amino acid residues was determined and was as predicted from the DNA . Binding of BlaI to DNA was detected by the slower migration of protein DNA complexes during polyacrylamide gel electrophoresis . BlaI was shown to selectively bind DNA fragments carrying the promoter regions of blaI and blaP.

FEBS Lett, 1987 Aug 10, 220(1), 43 - 6
The complete amino acid sequence of the ribosomal 'A' protein (L12) from Bacillus stearothermophilus; Garland WG et al.; The complete amino acid sequence of the ribosomal 'A' protein (Bst L12) has been determined from Bacillus stearothermophilus . The protein contains 122 amino acids and has a composition of Asp4, Ans3, Thr6, Glu20, Gln2, Pro3, Gly9, Ala23, Val13, Met2, Ile11, Leu8, Phe2, Lys15, Arg1 and a molecular mass of 12737 Da.

J Biol Chem, 1987 Aug 5, 262(22), 10672 - 7
Identification of highly reactive cysteinyl and methionyl residues of rabbit muscle phosphofructokinase; Latshaw SP et al.; The reactivity of the 16 thiol groups of rabbit skeletal muscle phosphofructokinase has been studied extensively over the past 20 years . Several of these thiols show high reactivity with a variety of reagents, display differential reactivity in the presence of allosteric ligands and substrates, and appear to be important to function because their modification changes activity and regulatory properties . In the present study, the location in the primary structure of several highly reactive thiol groups has been established by reaction with {14C}iodoacetate . In the course of these studies, 2 methionyl residues that are located at or near proposed ligand-binding sites are readily carboxymethylated by iodoacetate . In addition to confirming the presence of the most reactive thiol group at sequence position 88, a thiol protected from reaction by the presence of fructose-6-P and cyclic AMP has been found at position 169 . Cysteine 169 is close to a residue important to the binding of fructose-6-P in the homologous structure from Bacillus stearothermophilis phosphofructokinase . The modification of Cys-169 brings about extensive, but not total, loss of activity . Another cysteine, at position 232, was found to be highly reactive also . Substrate provided partial protection against carboxymethylation at this position . Carboxymethylation of enzyme restricted to methionines 74 and 173 brought about no changes in the total activity or in the ATP inhibition profile of the enzyme . This is significant since position 74 was projected on the basis of the homologous procaryotic structure to be important in the binding of nucleotide to the allosteric site.

Eur J Biochem, 1987 Aug 3, 166(3), 539 - 45
Structure and expression of genes coding for xylan-degrading enzymes of Bacillus pumilus; Moriyama H et al.; The complete nucleotide sequence of the beta-xylosidase gene (xynB) of Bacillus pumilus IPO and its flanking regions was established . A 1617-bp open reading frame for beta-xylosidase, a homodimer enzyme, was observed . The amino acid sequence of the N-terminal region and the molecular mass 62607 Da) of the beta-xylosidase subunit, deduced from the DNA sequence, agreed with the result obtained with the purified enzyme . The Shine-Dalgarno sequence was found 8 bp upstream of the initiation codon, ATG . The xylanase gene (xynA) of the same strain was 4.6 kbp downstream of the 3' end of xynB, and its DNA sequence was reported in our previous paper {Fukusaki, E., Panbangred, W., Shinmyo, A . & Okada, H . (1984) FEBS Lett . 171, 197-201} . The results of the Northern hybridization suggested that the mRNA of xynA and xynB were produced separately . The 5' and 3' ends of the xynA and xynB gene were mapped with nuclease S1 . The '-10' regions for promoter sequences of both genes were similar to the consensus sequence for B . subtilis RNA polymerases, the '-35' regions were different from all the known promoters for B . subtilis RNA polymerases.

Med Hypotheses, 1987 Aug, 23(4), 393 - 9
The effects of the delayed type immune response upon the host; Rivarola AJ; A consistent accumulation of experimental evidence, most of it published long ago, clearly shows that a constant consequence of the immune response elicited by the Koch bacillus is that the individual becomes susceptible to be injured or killed by the tuberculous antigen, which is totally innocuous to the virgin subject . This fact led Rich to conclude that throughout the natural history of tuberculosis, as soon as the immune reaction is activated, the tuberculo-protein poured out by the germ mass will exert its detrimental capabilities manifested as the constitutional symptoms of the disease . The pathogenicity of the specific antigen in the tuberculous-type immune reaction may be extrapolated to other states that elicit the same response, including cancer and tissue transplantation . Unexplained phenomena such as the general symptoms of many neoplasias, as well as the systemic manifestations of graft rejection, may then be attributed to the shedding of antigen from the foreign cell mass into the sensitized host.

J Urol, 1987 Aug, 138(2), 299 - 301
Intravesical bacillus Calmette-Guerin in the treatment of superficial transitional cell carcinoma of the bladder; Pansadoro V et al.; Pasteur strain bacillus Calmette-Guerin was used to treat 145 patients with superficial transitional cell carcinoma of the bladder: 47 had established residual disease (therapeutic group) and 130 received prophylactic/adjuvant therapy (including 32 who had a complete response in the therapeutic group and then were placed into the prophylactic group) . Among the patients in the therapeutic group a complete response rate of 68 per cent (32 of 47 patients, 95 per cent confidence limits 55 to 81 per cent) was achieved . Of those in the prophylactic/adjuvant group 85 per cent (111 of 130 patients, 95 per cent confidence limits 73 to 91 per cent) remain free of disease . The median followup for the therapeutic group was 17 months (range 3 to 49 months) . In the prophylactic/adjuvant therapy group the followup was greater than 3 years in 7 per cent, 2 to 3 years in 23 per cent, 1 to 2 years in 29 per cent and up to 1 year in 41 per cent (median 18 months) . Our study confirms that Pasteur strain bacillus Calmette-Guerin is safe and efficacious in the treatment and prevention of recurrent superficial bladder carcinoma.

Angew Parasitol, 1987 Aug, 28(3), 173 - 6
Insecticidal activity of Bacillus thuringiensis subspecies against Menopon gallinae (Mallophaga: Menoponidae); Lonc E et al.; 10 Bacillus thuringiensis strains were used to determine the susceptibility of Menopon gallinae (L) . In order to identify B . thuringiensis from dead insects, previously exposed to bacteria, the streptomycin-resistant mutants of strains were also applied . The experiments were conducted in a group of 20 specimens, in three replications . The absorbtion at 750 nm directly corresponded to the concentration of 23 X 10(6) spores/ml in each assay . The biting lice were exposed on agar plates, inoculated by 0.2 ml of B . thuringiensis preparations dipping for 2, 4 and 8 s or spraying with 0.5 ml of adequate strain . Laboratory trials showed that M . gallinae was susceptible to toxic activity of all tested strains . While the susceptibility to various serotypes varied, depending on the time and mode of infection, 3 subspecies: kurstaki, finitimus and kenyae were usually the most active . The least active strains were the subsp . galleriae and aizawi . The microbiological examination of dead mallophaga treated with B . thuringiensis proved the presence of this bacterium in smears taken from the insects . A comparison of morphological as well as biochemical properties of the antibiotics sensitive and the antibiotics-resistant mutants revealed no difference.

Protein Eng, 1987 Aug-Sep, 1(4), 319 - 25
Improvement in the alkaline stability of subtilisin using an efficient random mutagenesis and screening procedure; Cunningham BC et al.; An efficient random mutagenesis procedure coupled to a replica plate screen facilitated the isolation of mutant subtilisins from Bacillus amyloliquefaciens that had altered autolytic stability under alkaline conditions . Out of about 4000 clones screened, approximately 70 produced subtilisins with reduced stability (negatives) . Two clones produced a more stable subtilisin (positives) and were identified as having a single mutation, either Ile107Val or Lys213Arg (the wild-type amino acid is followed by the codon position and the mutant amino acid) . One of the negative mutants, Met50Val, was at a site where other homologous subtilisins contained a Phe . When the Met50Phe mutation was introduced into the B . amyloliquefaciens gene, the mutant subtilisin was more alkaline stable . The double mutant (Ile107Val/Lys213Arg) was more stable than the isolated single mutant parents . The triple mutant (Met50Phe/Ile107Val/Lys213Arg) was even more stable than Ile107Val/Lys213Arg (up to two times the autolytic half-time of wild-type at pH 12) . These studies demonstrate the feasibility for improving the alkaline stability of proteins by random mutagenesis and identifying potential sites where substitutions from homologous proteins can improve alkaline stability.

Biotechnol Appl Biochem, 1987 Aug, 9(4), 323 - 4
Thermally modified azocasein--a new insoluble substrate for the determination of proteolytic activity; Safarik I; An insoluble chromogenic substrate for the determination of proteolytic activity was prepared by heating azocasein in a thin layer at 200 degrees C for 4 h in a hot-air thermostat . The activity of an extracellular bacterial proteinase produced by Bacillus sp . was determined with this new substrate.

Scand J Immunol, 1987 Aug, 26(2), 149 - 59
Immunopurification of radiolabelled antigens of Mycobacterium leprae and Mycobacterium bovis (bacillus Calmette-Guerin) with monoclonal antibodies; Britton WJ et al.; Radiolabelled sonicate of Mycobacterium leprae when examined by SDS-PAGE and two-dimensional gel electrophoresis (2-DE) contained fewer antigens than the comparable sonicate from M . Bovis (bacillus Calmette-Guerin) (BCG) . A solid-phase immunopurification assay with anti-M . leprae monoclonal antibodies (MoAb) was used to characterize four of these antigens . Three of the MoAb were M . leprae-specific and with them antigens with apparent molecular weights (Mr) of 12,000 (12K), 18K, and 35K were isolated . On 2-DE, the heavily labelled 12K antigen was heterogeneous with a range in pI of 4.8-5.2 . The 35K antigen, which was identified by a conformational determinant, and the 18K antigen were also acidic proteins with pI of 5.4 and 5.1 . The fourth antigen was purified from both M . leprae and BCG sonicates and had an Mr of 70K and a pI of 5.1 . MoAb reacting with the cell wall protein of M . leprae resulted in separation of multiple bands ranging in Mr from 12K to 65K, rather than the dominant 65K protein seen in immunoblots . A similar pattern was obtained with MoAb that reacted with two cell wall polysaccharide antigens, and these antibodies may have co-precipitated the radiolabelled cell wall proteins . Immunoprecipitates of the M . leprae sonicate with human lepromatous leprosy sera, when analysed by 2-DE, were also found to contain the dominant 12K band and the 35K band . Furthermore, half the radiolabelled BCG antigens were precipitated by the same sera.

Proc Natl Acad Sci U S A, 1987 Aug, 84(15), 5167 - 71
Recruitment of substrate-specificity properties from one enzyme into a related one by protein engineering; Wells JA et al.; The Bacillus licheniformis and Bacillus amyloliquefaciens subtilisins differ by 31% in protein sequence and by factors of greater than 60 in catalytic efficiency, kcat/Km, toward various substrates . Despite large differences in sequence and substrate specificity for these serine proteases, only two amino acid substitutions (residues 156 and 217) occur within 4 A (contact distance) of modeled substrates, and a third substitution (residue 169) is within 7 A . The three B . licheniformis substitutions (Ser-156/Ala-169/Leu-217) were introduced into the wild-type B . amyloliquefaciens subtilisin (Glu-156/Gly-169/Tyr-217) by site-directed mutagenesis . The substrate specificity of the triple mutant approaches that of B . licheniformis enzyme when assayed with seven different substrates that vary in charge, size, and hydrophobicity . Thus, specificity properties of distantly related and functionally divergent enzymes can be exchanged by limited amino acid replacements, in this case representing less than 4% of the sequence differences.

J Urol, 1987 Aug, 138(2), 295 - 8
Single course versus maintenance bacillus Calmette-Guerin therapy for superficial bladder tumors: a prospective, randomized trial; Hudson MA et al.; A total of 42 patients with recurrent superficial bladder tumors or carcinoma in situ entered a prospective, randomized trial to compare the efficacy of bacillus Calmette-Guerin therapy with and without quarterly maintenance instillations of bacillus Calmette-Guerin . Maintenance therapy did not reduce further bladder tumor recurrence rates or the interval to recurrence in patients who responded to the initial course of therapy . However, prolongation of toxicity was observed with maintenance bacillus Calmette-Guerin therapy.

Biochimie, 1987 Aug, 69(8), 797 - 802
Baciphelacin: a new eukaryotic translation inhibitor; Carrasco L; Baciphelacin an antibiotic produced by Bacillus thiaminolyticus was a potent inhibitor of protein synthesis in HeLa cells and other mammalian cell lines . It had no effect on DNA or RNA synthesis . Concentrations of baciphelacin around 10(-7) M inhibited protein synthesis by 50% in intact cells . The antibiotic had no effect on protein synthesis in Saccharomyces cerevisiae or Escherichia coli, but inhibited the protozoan Trypanosoma brucei . In vitro protein synthesis in a rabbit reticulocyte cell-free system was blocked by baciphelacin . However, translation of globin mRNA in a wheat cell-free system was not affected by this antibiotic . Baciphelacin had no activity against a number of cell-free systems used to measure different steps of translation, including binding of substrates to the ribosome, peptide bond formation and polyphenylalanine synthesis . Therefore, it is assumed that it affects the initiation of translation or the charging of tRNA . Finally, the inhibition of protein synthesis by compounds structurally related to baciphelacin was tested and their effects compared to baciphelacin.

Bioorg Khim, 1987 Aug, 13(8), 1023 - 30
{Stepwise synthesis of oligonucleotides . XXXIV . Preparative synthesis of trinucleoside diphosphates and longer oligoribonucleotides using immobilized ribonucleases}; Zhenodarova SM et al.; Immobilized guanyl-specific ribonucleases from Aspergillus clavatus (C2), A . oryzae (T1), and Bacillus intermedius 7P (Bi) have been used for preparative synthesis of ten trinucleoside diphosphates, three tetra- and one pentanucleotide having the only guanylic acid residue at the 5'-end . The nucleotide sequence of the oligonucleotide synthesised determined the choice of the ribonuclease.

J R Soc Med, 1987 Aug, 80(8), 480 - 1
Pathogenic role of Bacillus cereus in wound infections in the tropics; Dryden MS; A bacteriological survey was undertaken on clinically infected traumatic wounds amongst a group of young and fit Operation Raleigh members, who were living and working in a remote area of Costa Rican rain forest . All infected wounds were swabbed before treatment and, where possible, at intervals during treatment . Swabs were also obtained from the nose and throat of each patient . All swabs were stored by desiccation in sterile silica gel for culture at a later date . Culture revealed a high rate of isolation of Bacillus cereus from the wounds . The organism was commonly isolated in pure and heavy growth . Contamination by B . cereus was considered and excluded experimentally . Preliminary toxological studies have shown that the majority of the isolates produce a necrotic exotoxin, in keeping with the clinical findings . These results suggest that B . cereus caused significant sepsis in this series of traumatic wounds.

Cancer, 1987 Aug 1, 60(3 Suppl), 635 - 44
Biologic response modifiers in genitourinary neoplasia; Droller MJ; With the exception of testis cancer, the variety of genitourinary cancers have not been found to be consistently responsive to chemotherapeutic agents or regimens for other than anecdotal short duration . This has generated keen interest in the possibility that biologic response modifiers might either directly or through manipulation of immune response mechanisms successfully prevent tumor progression in these systems . In recent years, those substances that have attracted the greatest attention have included interferon (and, more recently, recombinant gamma interferon), bacillus Calmette-Guerin (BCG), tumor necrosis factor, prostaglandin synthetase inhibitors, and interleukin-2 . Results with each of these agents in the variety of genitourinary cancers have been both promising and disappointing . A number of mechanisms have been suggested to underlie the actions of each of these substances, and the successful or unsuccessful recruitment of these mechanisms, in the context of the particular intrinsic behavior of the cancers being treated, have been suggested as reasons for the treatment results that have been seen . Therapeutic efficacy has been described in the treatment of renal cell cancer by both systemic interferon and interleukin-2 . Successful treatments have been reported in approximately 20% of patients treated with each substance, but generally, these results have been of short duration . Topical BCG has been used with great success to treat superficial transitional cell bladder cancer . In these instances, the generation of tumor necrosis factor has been suggested as possibly accounting for the 70% success rate both in therapy and prophylaxis that has been seen . Leukocyte-derived interferon and, more recently, recombinant gamma interferon, were found in initial trials to generate a 20% response rate in renal cell carcinoma patients . Enthusiasm for these agents, however, has been tempered more recently both by a failure to reproduce these results with any substantial duration as well as by the significant side effects that have been seen . Clearly, these agents continue to be intriguing both because of their intellectual appeal through the mechanisms by which they may be effective, as well as by the absence of any definitive therapy for the cancers they are being used to treat . An understanding of the complex host/tumor cell interaction that may ultimately determine therapeutic efficacy for each of these agents is undoubtedly critical if the role of these substances in the treatment of genitourinary cancer is to be successfully implemented, either alone or in combination with other treatment modalities.

Biochim Biophys Acta, 1987 Jul 31, 920(2), 155 - 60
Bacillus cereus phospholipase C: carboxylic acid ester specificity and stereoselectivity; Snyder WR; Thiophosphate analogs of phosphatidylcholine have been synthesized with varying structural complexity . These analogs have been used in a continuous spectrophotometric assay for phospholipase C (Bacillus cereus) to estimate the minimal structural requirements associated with the non-polar portion of the substrate phospholipid . The analogs were of three types containing zero, one or two carboxylic acid ester functionalities . The analogs with one or two ester groups acted as substrates for phospholipase C, while those without an ester functionality were not hydrolyzed . The rac-phosphatidylcholine analog with two ester functionalities gave biphasic time-course results, and was subsequently resolved into enantiomers by selective hydrolysis with a sterospecific phospholipase A2 (Crotalus atrox) . The enantiomer with R absolute configuration was rapidly hydrolyzed by the phospholipase C while the enantiomer with the S configuration was slowly hydrolyzed after a long induction period . The results suggest that the B . cereus phospholipase C is specific for an ester functionality and is stereoselective for the R absolute configuration at glycerol C-2.

J Biol Chem, 1987 Jul 25, 262(21), 10346 - 54
Novel phenylalanine dehydrogenases from Sporosarcina ureae and Bacillus sphaericus . Purification and characterization; Asano Y et al.; NAD+-dependent phenylalanine dehydrogenases were purified 1,500- and 1,600-fold, and crystallized from Sporosarcina ureae SCRC-R04 and Bacillus sphaericus SCRC-R79a, respectively . The purified enzymes were homogeneous as judged by disc gel electrophoresis . The enzyme from S . ureae has a molecular weight of 305,000, while that of B . sphaericus has a molecular weight of 340,000 . Each is probably composed of eight subunits identical in molecular weight . The S . ureae enzyme showed a high substrate specificity in the oxidative deamination reaction acting on L-phenylalanine, while that of B . sphaericus acted on L-phenylalanine and L-tyrosine . The enzymes had lower substrate specificities in the reductive amination reaction acting on alpha-keto acids . The Sporosarcina enzyme acted on phenylpyruvate, alpha-ketocaproate, alpha-keto-gamma-methylthiobutyrate and rho-hydroxyphenylpyruvate . The Bacillus enzyme acted on rho-hydroxyphenylpyruvate, phenylpyruvate, and alpha-keto-gamma-methylthiobutyrate . The enzyme from B . sphaericus catalyzes The enzyme from B . sphaericus catalyzes the transfer of pro-S (B) hydrogen from NADH.

Science, 1987 Jul 24, 237(4813), 394 - 9
Engineering enzyme specificity by "substrate-assisted catalysis"; Carter P et al.; A novel approach to engineering enzyme specificity is presented in which a catalytic group from an enzyme is first removed by site-directed mutagenesis causing inactivation . Activity is then partially restored by substrates containing the missing catalytic functional group . Replacement of the catalytic His with Ala in the Bacillus amyloliquefaciens subtilisin gene (the mutant is designated His64Ala) by site-directed mutagenesis reduces the catalytic efficiency (kcat/Km) by a factor of a million when assayed with N-succinyl-L-Phe-L-Ala-L-Ala-L-Phe-p-nitroanilide (sFAAF-pNA) . Model building studies showed that a His side chain at the P2 position of a substrate bound at the active site of subtilisin could be virtually superimposed on the catalytic His side chain of this serine protease . Accordingly, the His64Ala mutant hydrolyzes a His P2 substrate (sFAHF-pNA) up to 400 times faster than a homologous Ala P2 or Gln P2 substrate (sFAAF-pNA or sFAQF-pNA) at pH 8.0 . In contrast, the wild-type enzyme hydrolyzes these three substrates with similar catalytic efficiencies . Additional data from substrate-dependent pH profiles and hydrolysis of large polypeptides indicate that the His64Ala mutant enzyme can recover partially the function of the lost catalytic histidine from a His P2 side chain on the substrate . Such "substrate-assisted catalysis" provides a new basis for engineering enzymes with very narrow and potentially useful substrate specificities . These studies also suggest a possible functional intermediate in the evolution of the catalytic triad of serine proteases.

Clin Chim Acta, 1987 Jul 15, 166(2-3), 163 - 9
A simple enzymatic fluorimetric method for the determination of branched-chain L-amino acids in microlitre volumes of plasma; Gleeson M et al.; Existing methods for the estimation of the branched-chain amino acids (BCAA) normally require sophisticated, expensive instrumentation . An alternative is an enzymatic spectrophotometric method but this requires relatively large volumes of blood and is rather time-consuming . A simple enzymatic fluorimetric method for the measurement of the BCAA in microliter samples of plasma is described here . The method is based on the oxidative deamination of L-leucine, L-isoleucine and L-valine by leucine dehydrogenase from Bacillus species . The NADH which is formed in stoichiometric quantities is estimated fluorimetrically . In the presence of the ketone-trapping agent hydrazine the reaction goes to completion in an alkaline incubation medium at 37 degrees C . By this method the combined BCAA can be measured routinely in 20 microliter sample of plasma . The test exhibits acceptable precision and reproducibility.

Biochem Biophys Res Commun, 1987 Jul 15, 146(1), 346 - 53
The importance of arginine 171 in substrate binding by Bacillus stearothermophilus lactate dehydrogenase; Hart KW et al.; A variant of lactate dehydrogenase from Bacillus stearothermophilus has been engineered by site-directed mutagenesis in which an active-site arginine residue at position 171 in the protein sequence is replaced by lysine . Replacement of this arginine by lysine has no effect on co-enzyme binding, a relatively small effect on the rate of turnover of the enzyme, but causes a 2000-fold increase in the Michaelis constant for pyruvate, a 6000-fold increase in the dissociation constant for oxamate and results in a Michaelis constant for lactate which is too high to measure . The decrease in binding energy for these carboxylate-containing substrates caused by this mutation is very large, around 5.5 kcal.mol-1 and in part, is explained by the small increase in the distance of a lysine-substrate carboxylate interaction at this site and the absence of the additional hydrogen bond from a two-point arginine-carboxylate interaction . Consistent with this last observation, the ability of this mutant enzyme to stabilize an NAD+-sulphite compound in its active site (an alternative enzyme-substrate complex which does not involve bifurcated bonding to arginine) is only reduced 14-fold.

Biochem Biophys Res Commun, 1987 Jul 15, 146(1), 173 - 8
Synthesis of P1,P4-di(adenosine 5'-) tetraphosphate by leucyl-tRNA synthetase, coupled with ATP regeneration; Kitabatake S et al.; A simple and practical procedure for the synthesis of P1,P4-di(adenosine 5'-) tetraphosphate from ATP by the catalysis of leucyl-tRNA synthetase from Bacillus stearothermophilus is described . Km for leucine was 6.7 microM and for ATP was 3.3 mM . The reaction yielded not only diadenosine tetraphosphate, but various byproducts such as P1,P3-(diadenosine 5'-) triphosphate, ADP and AMP . By coupling the reaction with an ATP regeneration system by acetate kinase and adenylate kinase with acetylphosphate as a phosphate donor, diadenosine tetraphosphate was prepared as a sole product at a high yield (96%).

J Immunol, 1987 Jul 15, 139(2), 518 - 25
Induction of nitrite/nitrate synthesis in murine macrophages by BCG infection, lymphokines, or interferon-gamma; Stuehr DJ et al.; Macrophage synthesis of nitrite and nitrate after activation by BCG infection or by treatment in vitro with both T cell-derived (lymphokines (LK) or recombinant murine interferon-gamma (IFN-gamma} and bacterial (lipopolysaccharide (LPS) and heat-killed bacillus Calmette-Guerin (hk BCG} agents was studied by using macrophages from C3H/He and C3H/HeJ mice . Spleen and peritoneal macrophages isolated from BCG-infected donors that were producing nitrate continued to synthesize nitrite and nitrate in culture . LPS treatment in vitro (25 or 50 micrograms/ml) additionally increased this nitrite/nitrate synthesis . Thioglycolate-elicited macrophages from non-infected C3H/HeJ mice treated with LK also produced nitrite/nitrate, and concurrent LPS (0.1 to 50 micrograms/ml) treatment resulted in enhanced synthesis . Recombinant IFN-gamma also stimulated nitrite/nitrate synthesis by C3H/He and CeH/HeJ macrophages as did LPS (C3H/He only) and hk BCG . When given concurrently with either LPS or hk BCG, IFN-gamma enhanced C3H/He and C3H/HeJ macrophage nitrite/nitrate synthesis over that produced by macrophages treated with either LPS or hk BCG alone . Macrophages activated in vitro exhibited a 4 to 12 hr lag time before engaging in nitrite/nitrate synthesis, which then proceeded for 36 to 42 hr at linear rates . Daily medium renewal did not alter the synthesis kinetics but increased the total amount of nitrite/nitrate produced . Nitrate and nitrite were stable under the conditions of culture and when added did not influence additional macrophage synthesis . Taken together, these results indicate that T cell lymphokines and IFN-gamma are powerful modulators of macrophage nitrite/nitrate synthesis during BCG infection and in vitro, and nitrite/nitrate synthesis appears to be common property of both primed and fully activated macrophage populations.

Biochem J, 1987 Jul 15, 245(2), 467 - 72
Characterization of structural component of cell walls of alkalophilic strain of Bacillus sp . C-125 . Preparation of poly(gamma-L-glutamate) from cell wall component; Aono R; The cell wall of an alkalophilic strain of Bacillus sp . C-125 is composed of A1 gamma-peptidoglycan, a teichuronic acid and an unknown acidic polymer composed of glutamic acid and glucuronic acid, of which the molar ratio is approx . 4-5:1 . Poly(gamma-L-glutamate) was prepared from the acidic polymer by removal of almost all of the glucuronic residues with trifluoromethanesulphonic acid treatment and purified chromatographically . The Mr of the polyglutamate preparation was estimated to be 14,000 by gel chromatography, or 43,000 on the basis of the content of N-terminal acid residues . The acidic polymer found in the cell wall of the organism was concluded to be a polyglutamate substituted with (oligo)glucuronic acid residues or a complex composed of two kinds of polymers (polyglutamate and polyglucuronate).

Biochemistry, 1987 Jul 14, 26(14), 4248 - 58
Kinetic and structural characterization of reversibly inactivated beta-lactamase; Fink AL et al.; The reversible inhibition of beta-lactamase I from Bacillus cereus by cloxacillin, methicillin, and nafcillin has been systematically investigated . For these substrates the enzymatic reaction involves partitioning of the substrate between turnover and inhibition . Typically, concentrations of several hundred millimolar are necessary for complete inactivation . The completely inactivated enzyme could be formed by incubation at temperatures above 20 degrees C, where inhibition competes more effectively with turnover, and then stabilized by dropping the temperature to 0 degrees C or lower . The inactivated enzyme was rapidly separated from unreacted substrate and product at low temperature by centrifugal gel filtration or ion exchange and examined by far-UV circular dichroism for evidence of a conformational change . At pH 7 the inactivated enzyme had a secondary structure essentially identical with that of the native enzyme . The fluorescence emission spectrum of the inactivated enzyme (at pH 7) was also identical with that of the native enzyme . However, the inactivated enzyme was found to be considerably more sensitive to thermal denaturation, to acid-induced conformational isomerization, and to trypsinolysis than the native enzyme . We conclude from the circular dichroism results that the structure of the reversibly inactivated enzyme cannot be significantly different from that of the native enzyme . Therefore, previous findings that have been interpreted as indicating a major conformational change must be reevaluated . From examination of the low-resolution crystallographic structure of the enzyme we propose that the most likely cause of the inactivation is an alternate conformational state of the acyl-enzyme intermediate involving movement of one or more of the alpha-helices forming part of the active site . Such a structural effect could leave the secondary structure unchanged but have significant effects on the tertiary structure, catalysis, mobility, and susceptibility to trypsin and denaturation . We propose that the underlying physical reason why certain beta-lactam substrates bring about this "substrate-induced deactivation", or suicide inactivation, of the enzyme is due to the presence of the alternative acyl-enzyme conformation of similar free energy to the productive one, in which one (or more) essential catalytic group is no longer optimally oriented for catalyzing deacylation . Thus for substrates with a slow rate of deacylation (less than or equal to 100 s-1) the conformational transition can compete effectively on the time scale of the turnover reaction.

Nucleic Acids Res, 1987 Jul 10, 15(13), 5251 - 9
Nucleotide sequence of Bacillus phage Nf terminal protein gene; Leavitt MC et al.; The nucleotide sequence of Bacillus phage Nf gene E has been determined . Gene E codes for phage terminal protein which is the primer necessary for the initiation of DNA replication . The deduced amino acid sequence of Nf terminal protein is approximately 66% homologous with the terminal proteins of Bacillus phages PZA and luminal diameter 29, and shows similar hydropathy and secondary structure predictions . A serine which has been identified as the residue which covalently links the protein to the 5' end of the genome in luminal diameter 29, is conserved in all three phages . The hydropathic and secondary structural environment of this serine is similar in these phage terminal proteins and also similar to the linking serine of adenovirus terminal protein.

Jikken Dobutsu, 1987 Jul, 36(3), 239 - 44
Inactivation of Bacillus piliformis spores by heat and certain chemical disinfectants; Itoh T et al.; The inactivation of Tyzzer's organism (Bacillus piliformis) spore isolated from rats by heat and various chemical disinfectants was studied . The spores were from B . piliformis-infected rat liver tissues . The spore suspension (10(4) 50% of rat liver lesion producing dose with prednisolone treatment/ml) was treated with heart or disinfectants . Inactivation of the spores was examined in experimentally infected rats . Rats were inoculated perorally with a treated spore suspension and injected subcutaneously with prednisolone . On the sixth day after inoculation, rats were examined grossly for liver lesions . Spores were inactivated at 80 degrees C for 15 min but not at 60 degrees C for 30 min . Spores were inactivated by 0.4% peracetic acid, 0.015% sodium hypochrolite, 1% iodophol, 5% phenol . Alcide and 0.37% formaldehyde solution, but not by 0.037% formaldehyde solution, 70% ethanol, 0.3% benzethonium chloride solution, 3% cresol and soap solution, or 4% chlorhexidine digluconate . These findings suggest that B . piliformis spores are relatively sensitive to heat and certain chemical disinfectants.

Aust Vet J, 1987 Jul, 64(7), 207 - 10
Natural establishment of thiaminase activity in the alimentary tract of newborn lambs and effects on thiamine status and growth rates; Thomas KW et al.; Thiaminase activity was detected in the faeces of lambs at 2 to 5 days of age . Levels of activity increased for 10 days and then declined over the next 3 to 4 weeks . Decreased erythrocyte transketolase activity indicated thiamine insufficiency in lambs with high thiaminase activity . Mean growth rates were 17% less in lambs with high thiaminase activity than in lambs with zero or low thiaminase activity . Bacillus thiaminolyticus was the only organism isolated which produced thiaminase . Treatment of newborn lambs with intramuscular injections of sulphadoxine did not prevent them from excreting thiaminase in their faeces . It is proposed that oral thiamine supplementation of lambs at 2 to 3 weeks of age may be the most appropriate prevention and treatment for subclinical thiamine deficiency of the cause described.

Ann Inst Pasteur Microbiol, 1987 Jul-Aug, 138(4), 471 - 84
Ultrastructural midgut events in Culicidae larvae fed with Bacillus sphaericus 2297 spore/crystal complex; Charles JF; Ingestion of Bacillus sphaericus 2297 spore/crystal complex by Culicidae larvae Anopheles stephensi, Culex pipiens subsp . pipiens and Aedes aegypti was rapidly followed by a dissolution of the protein crystalline inclusions inside the anterior stomach of the three species . During the first day of intoxication, B . sphaericus spores germinated within the midgut lumen, and were in a vegetative stage between 36-48 h after ingestion when the larvae began to die . Ultrastructural observations focused on larval midgut showed alterations which differed according to the mosquito species, being localized mainly in the gastric caeca and posterior stomach . With the bacterial concentration used, neither general cell swelling nor complete breakdown of the midgut epithelium was recorded before larval death . In A . stephensi larval midgut epithelium large low-electron-density areas appeared, rough endoplasmic reticula formed numerous concentrical structures and mitochondria swelled . Large vacuoles (of unknown origin) appeared early on in the C . pipiens midgut cells, and rough endoplasmic reticula broke into small vesicles . Midgut epithelial cells of A . aegypti showed mitochondria swelling except in the anterior stomach, and a vacuolisation of smooth reticula: these aspects remained unchanged until the larvae died.

Ann Inst Pasteur Microbiol, 1987 Jul-Aug, 138(4), 457 - 70
Similarities between crystal protein subunits of Bacillus thuringiensis strain 1884 serotype H14 and strain PG14 serotype H8a,8b, and their relationship with mosquitocidal activity; Thiery I; Bacillus thuringiensis subsp . israelensis strain 1884 and B . thuringiensis subsp . morrisoni strain PG14 have the same toxicity toward mosquito larvae . Their protein crystal contents were compared either on native electrophoretic systems or on gel filtration . Analysis of electroeluted native proteins in SDS-PAGE showed that the 28-Kd protein alone, without the 68- or 130-Kd protein, was not larvicidal; however, toxicity was recorded as soon as 28- and 68-Kd proteins were associated . After gel filtration of 1884 soluble crystal, the 68-Kd protein alone, without 28 or 130 Kd, was not toxic; toxicity was also recorded when 28- and 68-Kd proteins and higher molecular weight proteins were associated . A comparable pattern was observed with PG14 soluble crystals.

Food Addit Contam, 1987 Jul-Sep, 4(3), 297 - 307
Excretion of oxytetracycline in eggs after medication of laying hens; Roudaut B et al.; The kinetics of oxytetracycline elimination into eggs were determined separately for albumen and yolk after oral administration through either drinking water (0.1-0.25 and 0.5 g/l for 5 days) or feed (300 and 600 ppm for 7 days) or after intramuscular injections (3 X 15 mg/kg body weight and 3 X 30 mg/kg body weight), 24 hours apart . Residues were assayed by a microbiological agar diffusion method, with Bacillus cereus as test-organism . The detection threshold was 0.07 micrograms/g for albumen and 0.2 micrograms/g for yolk . In all cases, the elimination period lasted longer for the yolk; it varied between 0 and 10 days after treatment was discontinued, according to administration routes and dosages . The conditions of oxytetracycline utilization in laying hens are discussed . The oral route only might be used to adhere to the proposals presented by the Joint FAO/WHO Expert Committee on Food Additives.

Am J Trop Med Hyg, 1987 Jul, 37(1), 174 - 9
The ultrastructure of actinomycetoma grains caused by Streptomyces somaliensis; Nasher M et al.; This study compares the in vitro ultrastructure of Streptomyces somaliensis with the grain formation seen in vivo . Cultured forms were cocco-bacillary with thick cell walls and septa . Individual cells stained with PAS.TCH and Alcian blue . Grains were largely composed of an electron-dense fibrillar matrix surrounding clear areas, some of which contained organisms . The ultrastructural appearances of the latter were not significantly different from those seen in vitro although the cell wall was less well defined and did not stain with Alcian blue, and there were more intracellular lipid droplets . Host cells surrounding the mass were incorporated into the structure of the grain, a process associated with cell death . The grains of S . somaliensis appear to be derived from both host and actinomycete elements . Compared to other mycetoma agents the mechanism of grain formation by this organism is distinctive.

J Bacteriol, 1987 Jul, 169(7), 3281 - 8
Structural disulfide bonds in the Bacillus thuringiensis subsp . israelensis protein crystal; Couche GA et al.; We examined disulfide bonds in mosquito larvicidal crystals produced by Bacillus thuringiensis subsp . israelensis . Intact crystals contained 2.01 X 10(-8) mol of free sulfhydryls and 3.24 X 10(-8) mol of disulfides per mg of protein . Reduced samples of alkali-solubilized crystals resolved into several proteins, the most prominent having apparent molecular sizes of 28, 70, 135, and 140 kilodaltons (kDa) . Nonreduced samples contained two new proteins of 52 and 26 kDa . When reduced, both the 52- and 26-kDa proteins were converted to 28-kDa proteins . Furthermore, both bands reacted with antiserum prepared against reduced 28-kDa protein . Approximately 50% of the crystal proteins could be solubilized without disulfide cleavage . These proteins were 70 kDa or smaller . Solubilization of the 135- and 140-kDa proteins required disulfide cleavage . Incubation of crystals at pH 12.0 for 2 h cleaved 40% of the disulfide bonds and solubilized 83% of the crystal protein . Alkali-stable disulfides were present in both the soluble and insoluble portions . The insoluble pellet contained 12 to 14 disulfides per 100 kDa of protein and was devoid of sulfhydryl groups . Alkali-solubilized proteins contained both intrachain and interchain disulfide bonds . Despite their structural significance, it is unlikely that disulfide bonds are involved in the formation or release of the larvicidal toxin.

Indian J Lepr, 1987 Jul-Sep, 59(3), 300 - 8
A survey on proportion of multibacillary cases of leprosy in Himachal Pradesh; Dharmshaktu NS et al.; The present village survey indicates that though the percentage of Multi-bacillary cases remains high (50%) even after deducting the cases fit for discharge yet there is no evidence of high proportion of MB cases among newly detected cases . The percentage of MB cases among newly detected Leprosy cases is 16.7, but when old and new cases are put together and cases fit for discharge are deducted-the percentage of MB cases increases to 50 . This high percentage of MB case is due to prolonged irregular treatment of old cases that are still clinically active, even after 10-30 years of treatment . The State level, district level/Leprosy Control Unit level data also indicated high percentage of MB cases which was mainly due to underdetection of cases particularly of Pauci-bacillary type, non-discharge of Multi-bacillary cases fit for discharge, prolonged irregular treatment of remaining MB cases that are active and due to various other contributory factors.

Mikrobiyol Bul, 1987 Jul, 21(3), 223 - 31
{The isolation, classification and some physiological properties of a protease-producing bacterium}; Cihangir N et al.; A proteolytic bacterium was isolated from specimen collected from soil . The protease producing bacterium showed optimal growth in media with neutral pH and when incubated at 30 degrees C aerobically . According to the classification results, the gram (+), sporulating bacillus was a Bacillus genus member . Also some physiological properties of the isolated bacterium was determined.

Mol Microbiol, 1987 Jul, 1(1), 59 - 66
Characterization of the toxicity and cytopathic specificity of a cloned Bacillus thuringiensis crystal protein using insect cell culture; Haider MZ et al.; An insecticidal protein gene from Bacillus thuringiensis var . aizawai was cloned in Escherichia coli . The cloned gene expressed at a high level and the synthesized protein appeared as an insoluble, phase-bright inclusion in the cytoplasm . These inclusions were isolated by density gradient centrifugation, the isolated protein was activated in vitro by different proteolytic regimes and the toxicity of the resulting preparations was studied using insect cells grown in tissue culture . The inclusions consisted of a 130 kDa polypeptide which was processed to a protease-resistant 55 kDa protein by tryptic digestion . This preparation lysed lepidopteran (Choristoneura fumiferana) CF1 cells but not dipteran (Aedes albopictus) cells . When the crystal protein was activated by sequential treatment, first with trypsin and then with Aedes aegypti gut proteases, the resulting 53 kDa polypeptide was now toxic only to the dipteran cells and not to the lepidopteran cells . Thus the dual specificity of this var . aizawai toxin results from differential proteolytic processing of a single protoxin . The trypsin-activated preparation was weakly active against Spodoptera frugiperda cells . Membrane binding studies of the trypsin-activated toxin revealed a 68 kDa protein in the lepidopteran cell membranes, which may be the receptor for this toxin.

Rev Infect Dis, 1987 Jul-Aug, 9(4), 693 - 703
Mixed bacterial meningitis; Downs NJ et al.; Two recent cases of mixed bacterial meningitis at the Kansas City Veterans Administration Medical Center were studied . A review of the literature suggests that 1% of all cases of meningitis are caused by more than one bacterial species . Before 1950 such cases occurred predominantly in children and were caused by combinations of bacteria commonly associated with meningitis . Since 1950 a largely adult population has been affected by mixed bacterial meningitis, with a higher incidence of gram-negative bacillary organisms cultured from the cerebrospinal fluid . Common predisposing factors in this older group of patients include infection at contiguous foci, tumors in close proximity to the central nervous system, or fistulous communications with the central nervous system . Mortality was 26% for cases occurring before 1950 and 63% for those occurring after 1950 . Failure to recognize one of the organisms present in the cerebrospinal fluid may result in the initiation of inadequate therapy in as many as 67% of cases . Empiric broad-spectrum antimicrobial therapy is indicated in symptomatic patients predisposed to mixed bacterial meningitis until culture results become available.

Appl Environ Microbiol, 1987 Jul, 53(7), 1701 - 4
Degradation of the framework of the Chlamydomonas cell wall by proteases present in a commercially available alpha-amylase preparation; Imam SH et al.; A commercially available alpha-amylase derived from Bacillus licheniformis contained an enzymatic activity able to degrade the inner portion or framework of the cell wall of Chlamydomonas reinhardtii . Both the wall-degrading activity and the contaminating protease were destroyed by heating the alpha-amylase preparation at 90 degrees C for 30 min . Since the alpha-amylase activity was uneffected by heat treatment, we conclude that it was not the alpha-amylase but the contaminating protease in the preparation that was responsible for the cell wall-degrading activity.

Am J Pathol, 1987 Jul, 128(1), 171 - 80
Endogenous peroxidase activity as a marker of macrophage renewal during BCG-induced inflammation in the rat lung; Warnock ML et al.; To determine whether the cytochemical localization of peroxidase activity could be used as a marker of monocyte influx into the lung during an inflammatory response, the authors studied the peroxidase phenotypes of lavaged alveolar macrophages from rats with bacille Calmette-Guerin (BCG)-induced pulmonary inflammation . Rats were immunized subcutaneously and 2 weeks later intravenously with BCG . During the early phase of pulmonary inflammation, an increase was observed in the numbers of alveolar macrophages with no peroxidase activity in the endoplasmic reticulum . These cells appeared to reflect monocyte influx into the injured lung . The later stages of inflammation were characterized by increased numbers of alveolar macrophages with peroxidase-positive endoplasmic reticulum, probably due to activation of enzymatic activity in situ . During the early phase, peroxidase activity was also observed within macrophage cytoplasmic inclusions, probably representing both primary monocyte lysosomes and internalized myeloperoxidase from inflammatory neutrophils . Serial observations indicated that the peroxidase-positive cytoplasmic inclusions became negative with time . It is concluded that inflammation-induced modulation of peroxidase activity in the endoplasmic reticulum and in cytoplasmic inclusions makes the alveolar macrophage peroxidase phenotype no more than a rough marker of monocyte influx into the inflamed lung.

Infect Immun, 1987 Jul, 55(7), 1707 - 14
Mycobacterium bovis BCG-induced protection against cutaneous and systemic Leishmania major infections of mice; Fortier AH et al.; We examined the protective effects of Mycobacterium bovis bacillus Calmette-Guerin (BCG) administration on Leishmania major infections of BALB/c and P/J mice . There were two treatment protocols . In the first, the footpads of naive animals were inoculated with mixtures of L . major and BCG (viable or heat killed) or the soluble mycobacterial antigen, purified protein derivative . Viable BCG, but not heat-killed BCG or purified protein derivative, inoculated with L . major amastigotes into the footpads of naive BALB/c or P/J mice protected these animals from the metastatic spread of parasites to the viscera and from ensuing lethal systemic infection . This treatment also induced cures of the cutaneous lesions of P/J mice but not of BALB/c mice . In the second protocol, we induced an immune response to BCG before inoculation of L . major . BCG given intraperitoneally 10 days before infection of footpads with leishmania offered protection against the metastatic spread of amastigotes in both P/J and BALB/c mice, regardless of intralesional treatment, and modulated the severity of cutaneous infection by 30 to 50% . Inoculation of a mixture of viable BCG and L . major amastigotes into BCG-immune mice completely protected both BALB/c and P/J strains from cutaneous disease; we recovered no parasites from the inoculated footpads of these animals . Furthermore, each of the nonspecifically protected mice of both the BALB/c and P/J strains developed immunity to rechallenge with viable L . major . Injection of amastigotes at a site remote from the original lesion, the contralateral footpad, resulted in the complete clearance of parasites in the inoculum with no evidence of either cutaneous or systemic disease over an extended observation period.

Mikrobiologiia, 1987 Jul-Aug, 56(4), 558 - 63
{The role of molecular oxygen and energy metabolism in the onset of Bacillus cereus spore germination}; Pronin SV; Bacillus cereus 96 spore germination was shown to depend on the content of molecular oxygen in the growth medium . When oxygen was removed from the medium, the spores germinated 50 min later as compared with this process under aerobic conditions . Likewise, spore initiation was delayed by 50 min in a growth medium containing oxygen in quantities optimal for respiration if 100mM KCN was added to it . The spores did not germinate when they had been treated simultaneously with glycolysis and respiration inhibitors.

Anal Biochem, 1987 Jul, 164(1), 199 - 206
A continuous spectrophotometric assay for the Bacillus cereus phospholipase C using a thiophosphate substrate analog: evaluation of assay conditions and chromogenic agents; Snyder WR; A thiophosphate analog of dioctanoylphosphatidylcholine has been used as the substrate in a continuous spectrophotometric assay for the Bacillus cereus phospholipase C . The reaction has been monitored at 412 nm using 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and at 324 nm using 4,4'-dithiopuridine (DTP) as the respective thiol-reactive chromogenic agents . An optimum pH 6.0 was determined for the phospholipase C-catalyzed reaction which was independent of the chromogen utilized . Although the reaction rates observed when DTP was used were increased over those seen with DTNB, the rates were insensitive to changes in the concentration of the chromogen normally used for the assay . The initial velocities were shown to be linearly dependent upon the amount of enzyme added over at least a 20-fold enzyme concentration range . The dependency of the initial velocity on the concentration of substrate showed a discontinuity at {S} = 40 microM when either DTP or DTNB was used . This was consistent with a value of 56 microM estimated for the substrate critical micelle concentration by an independent measurement . While the substrate data measured using DTP could not be fit to existing equations based on Michaelis-Menten kinetics, the data obtained using DTNB as the chromogen conformed with the model proposed by Wells for enzymes acting upon micelle-forming substrates (M . A . Wells (1974, Biochemistry 13, 2248-2257) . This allowed for the estimation of monomer and micelle Michaelis-Menten parameters for the B . cereus phospholipase C-catalyzed reaction with a thiophosphate analog substrate.

J Bacteriol, 1987 Jul, 169(7), 3088 - 93
Cloning and nucleotide sequencing of genes for a second type of small, acid-soluble spore proteins of Bacillus cereus, Bacillus stearothermophilus, and "Thermoactinomyces thalpophilus"; Sun DX et al.; The nucleotide sequences of the single genes coding for the B-type small, acid-soluble spore proteins (SASP) of Bacillus cereus, B . stearothermophilus, and "Thermoactinomyces thalpophilus" were determined, and the amino acid sequences of all B-type SASP were compared . While this type of SASP showed significant sequence conservation around the two spore protease cleavage sites, alignment of these sequences required the introduction of gaps, and even then only 19 of the residues were conserved exactly in all five proteins . However, all five B-type SASP did contain a large (27 to 35-residue), rather well-conserved amino acid sequence repeat, and four of the five proteins had well-conserved regions of 14 to 17 amino acids which appeared three times.

Z Naturforsch {C}, 1987 Jul-Aug, 42(7-8), 907 - 15
Reversible pH-induced dissociation of glucose dehydrogenase from Bacillus megaterium . II . Kinetics and mechanism; Maurer E et al.; Glucose dehydrogenase from Bacillus megaterium exists as a stable, active tetramer at pH 6.5 . By shifting the pH to 9, the enzyme is, completely and reversibly, dissociated into four inactive protomers . Kinetics and mechanism of this pH-induced dissociation have been studied, at various enzyme concentrations, by ultraviolet absorption, circular dichroism, normal and stopped-flow fluorescence as well as by light scattering and activity measurements . Dissociation of the fully active tetramer proceeds via three distinct kinetic steps: (1) fast conformational rearrangement of the tetramer, without any loss of activity (t1/2 0.0075 sec); (2) slow isomerization to a tetramer with lower specific activity (t1/2 27 sec); (3) subsequent dissociation of this rearranged tetramer into inactive monomers (t1/2 114 sec) with still intact native secondary structure . All three processes follow first-order kinetics . Both rate and extent of the dissociation are reduced, with a concomitant shift to higher reaction orders, by increasing the NaCl concentration in the buffer . This suggests the establishment of a dissociation/association equilibrium, due to the concentration-dependent stabilization of the tetrameric enzyme state by NaCl.

Mol Gen Genet, 1987 Jul, 208(3), 384 - 9
Cloning and expression of 130-kd mosquito-larvicidal delta-endotoxin gene of Bacillus thuringiensis var . Israelensis in Escherichia coli; Angsuthanasombat C et al.; Five recombinant E . coli clones exhibiting toxicity to Aedes aegypti larvae were obtained from a library of 800 clones containing XbaI DNA fragments of 110 kb plasmid from B . thuringiensis var . israelensis . All the five clones (pMU 14/258/303/388/679) had the same 3.8-kb insert and encoded a major protein of 130 kDa which was highly toxic to A . aegypti larvae . Three clones (pMU 258/303/388) transcribed the 130 kD a gene in the same direction as that of lac Z promoter of pUC12 vector whereas the transcription of the other two (pMU 14/679) was in the opposite direction . A 1.9-kb fragment of the 3.8 kb insert coded for a protein of 65 kDa . Partial DNA sequence of the 3.8 kb insert, corresponding to the 5'-terminal of the 130 kDa gene, revealed a continuous reading frame, a Shine-Dalgarno sequence and a tentative 5'-regulatory region . These results demonstrated that the 3.8 kb insert is a minimal DNA fragment containing a regulatory region plus the coding sequence of the 130 kDa protein that is highly toxic to mosquito larvae.

Appl Environ Microbiol, 1987 Jul, 53(7), 1525 - 30
Transfer of chromosomal genes and plasmids in Bacillus thuringiensis; Aronson AI et al.; A low frequency of chromosomal gene transfer from Bacillus thuringiensis to Bacillus cereus was detected by cell mating, with a tryptophan marker being the most frequently transferred gene among four that were tested . The process was resistant to DNase and was not mediated by cell filtrates . Among several B . thuringiensis subspecies tested, transfer was best with a derivative of B . thuringiensis subsp . kurstaki HD1, which lost several plasmids . All of the B . cereus recombinants contained at least one plasmid from the donor B . thuringiensis; frequently, it was a plasmid that encoded a protoxin gene . In matings with B . thuringiensis subsp . kurstaki HD1, a 29-megadalton plasmid that contained a ca . 2.5-kilobase region of homology with the chromosome was always transferred . No detectable transfer of chromosomal genes was found in B . thuringiensis subsp . kurstaki HD1 strains lacking this plasmid, suggesting that there may be chromosome mobilization.

Mol Microbiol, 1987 Jul, 1(1), 86 - 91
Molecular cloning and characterization of the beta-amylase gene from Bacillus circulans; Siggens KW; A gene encoding the beta-amylase of Bacillus circulans was isolated from a lambda library and sequenced . The structural gene consists of a 1725 bp open reading frame encoding a polypeptide with a predicted molecular wt of 62830 Daltons . Two active forms of the enzyme were found when the gene was expressed in E . coli . The larger 60 kD form was approximately 3 kD larger than the mature beta-amylase secreted from B . circulans, suggesting that processing of this protein is different between the two species . The smaller 49 kD form is also present at a low level in B . circulans and may result from proteolytic cleavage . The enzyme has a temperature optimum of 50 degrees C . Two other genes, one encoding an alpha-amylase and one a pullulanase, were also isolated from the lambda library.

Am J Med, 1987 Jul, 83(1), 165 - 70
Periodic acid-Schiff-negative granulomatous lymphadenopathy in patients with Whipple's disease . Localization of the Whipple bacillus to noncaseating granulomas by electron microscopy; Wilcox GM et al.; The diagnosis of Whipple's disease in a 58-year-old man was based on the finding of periodic acid-Schiff (PAS)-positive foamy macrophages on duodenal biopsy and demonstration of the typical bacilliform bodies by electron microscopy . The patient also had generalized peripheral lymphadenopathy with lymph node biopsy showing PAS-negative noncaseating granulomas . Electron microscopic examination of the lymph node specimen demonstrated a small number of typical bacilliform bodies with localization specifically to the granulomas in the lymph node . This finding of bacilliform bodies within PAS-negative noncaseating granulomas has not been reported previously . Localization of the Whipple bacillus specifically to noncaseating granulomas suggests that some patients with the disease may manifest a delayed hypersensitivity reaction to the bacillus.

Biochemistry, 1987 Jun 30, 26(13), 4131 - 8
Effects of engineering complementary charged residues into the hydrophobic subunit interface of tyrosyl-tRNA synthetase . Appendix: Kinetic analysis of dimeric enzymes that reversibly dissociate into inactive subunits; Ward WH et al.; Wild-type tyrosyl-tRNA synthetase (TyrTS) from Bacillus stearothermophilus is a symmetrical dimer . Four different heterodimeric enzymes have been produced by site-directed mutagenesis at the subunit interface so that the monomers are linked by a potential salt bridge in a hydrophobic environment . The two Phe-164 residues of wild-type TyrTS are on the axis of symmetry and interact in a hydrophobic region of the subunit interface . Mutation of Phe-164 to aspartate or glutamate in full-length TyrTS and to lysine or arginine in an active truncated enzyme (delta TyrTS) induces reversible dissociation of the enzyme into inactive monomers . Mixing mutants in equimolar amounts produces four different heterodimers: TyrTS(Asp-164)-delta TyrTS(Lys-164); TyrTS(Asp-164)-delta TyrTS(Arg-164); TyrTS(Glu-164)-delta TyrTS(Lys-164); TyrTS(Glu-164)-delta TyrTS(Arg-164) . A general method is derived for analyzing the kinetics of dimeric enzymes that reversibly dissociate into inactive subunits . Application to mutants of TyrTS allows estimation of dissociation constants (Kd values) for the dimers . At pH 7.8, the heterodimers have Kd values of 6-14 microM, whereas for homodimers Kd = 120-4000 microM . These values decrease to about 30 microM for homodimers of TyrTS(Asp-164), TyrTS(Glu-164), and delta TyrTS(Lys-164) when the pH favors uncharged forms of the side chains at position 164 . Each of the four salt bridges engineered into the hydrophobic subunit interface of TyrTS appears, therefore, to be weak . These engineered salt bridges may be compared with naturally occurring ones . In the latter, there are complementary interactions between the charges in the salt bridge with polar groups in the protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Med, 1987 Jun 26, 82(6B), 40 - 6
Norfloxacin for prevention of bacterial infections in granulocytopenic patients; Winston DJ et al.; The efficacy and safety of norfloxacin were compared with those of placebo, vancomycin-polymyxin, and trimethoprim-sulfamethoxazole (TMP/SMX) for prophylaxis of bacterial infections in granulocytopenic patients . The study results showed that norfloxacin treatment, which was well tolerated and not associated with any serious systemic adverse effects, prevented acquisition of gram-negative bacillary organisms . Fewer norfloxacin-treated patients (38 of 108 patients, or 35 percent) experienced microbiologically documented infections compared with patients receiving placebo (27 of 40 patients, or 68 percent), vancomycin-polymyxin (16 of 30 patients, or 53 percent), or TMP/SMX (14 of 28 patients, or 50 percent) . Gram-negative bacteremia developed in five of 108 norfloxacin-treated patients (5 percent), compared with 17 of 40 placebo-treated patients (43 percent), five of 30 treated with vancomycin-polymyxin (17 percent), and one of 28 patients treated with TMP/SMX (4 percent) . The incidence of gram-positive bacteremia was similar in all study groups and was not affected by norfloxacin or any other oral prophylactic antibiotics . These results suggest that norfloxacin is both safe and effective for the prevention of serious gram-negative bacillary infections in granulocytopenic patients . More effective prophylaxis of gram-positive bacterial infections, however, is needed.

Biochim Biophys Acta, 1987 Jun 22, 924(3), 467 - 72
Gluconate metabolism in germinated spores of Bacillus megaterium QM B1551: primary roles of gluconokinase and the pentose cycle; Otani M et al.; The metabolic pathway of gluconate, a major product of glucose metabolism during spore germination, was investigated in Bacillus megaterium QM B1551 . Compared to the parent, mutant spores lacking gluconokinase could not metabolize gluconate, whereas the revertant simultaneously restored the enzyme activity and the ability to metabolize it, indicating that gluconokinase was solely responsible for the onset of gluconate metabolism . To identify a further metabolic route for gluconate, we determined 14C yields in acetate and CO2 formed from {14C}gluconate, and found that experimental ratios of 14CO2/{14C}acetate obtained from {2-14C}gluconate and {3,4-14C}gluconate were not compatible with the ratios predicted from the Entner-Doudoroff pathway . In contrast, when CO2 release caused by recycling (approx . 30%) was corrected, the ratios almost agreed with those from the pentose cycle . Comparison of specific radioactivities in acetate also supported the conclusion that gluconate was metabolized via the pentose cycle, subsequently metabolized via the Embden-Meyerhof pathway, and finally degraded to acetate and CO2 without a contribution by the Krebs cycle.

Biochemistry, 1987 Jun 16, 26(12), 3417 - 25
1-Aminocyclopropanephosphonate: time-dependent inactivation of 1-aminocyclopropanecarboxylate deaminase and Bacillus stearothermophilus alanine racemase by slow dissociation behavior; Erion MD et al.; 1-Aminocyclopropanephosphonate (ACPP) was synthesized, and its effects on the pyridoxal 5'-phosphate linked enzymes 1-aminocyclopropanecarboxylate (ACPC) deaminase from Pseudomonas sp . ACPC and alanine racemase from Bacillus stearothermophilus were studied . ACPP was found to be a potent inhibitor of both enzymes with Km/Ki ratios of 500 and 2000, respectively . Inhibition for both enzymes was characterized by slow-binding (second-order rate constants less than 150 M-1 s-1) slow-dissociating behavior . Analysis of the pre-steady-state kinetics revealed a kinetically detectable intermediate E.I complex in the inhibition mechanism for the racemase but not for the deaminase . The one-step deaminase inhibition (Formula: see text) mechanism had an association rate constant (k1) of 100 M-1 s-1, a value 10(6)-fold slower than diffusion, suggesting either a slow alignment of the inhibitor at the enzyme active site or, more likely, the same mechanism as followed by racemase but with an E.I to E.I conversion rate (k3) that is sufficiently fast on the steady-state time scale so as to hinder detection of the initial weakly associated E.I intermediate . The E to E.I transition for the deaminase was further monitored by ultraviolet-visible and circular dichroism (CD) spectroscopies and found to exhibit a time-dependent shift in the visible absorption spectrum lambda max from 418 nm for the native enzyme to 333 nm at steady state, again consistent with a rapid E to E.I and slow E.I to E.I behavior . A rate constant for the absorbance shift of 150 M-1 s-1 was consistent with the k1 calculated in the inhibition studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochemistry, 1987 Jun 16, 26(12), 3443 - 6
Amino acid sequence at the citrate allosteric site of rabbit muscle phosphofructokinase; Kemp RG et al.; Previously, this laboratory has demonstrated {Colombo, G., & Kemp, R . G . (1976) Biochemistry 15, 1774-1780} that under appropriate conditions the citrate inhibitory binding site of rabbit skeletal muscle phosphofructokinase can be covalently modified by using pyridoxal phosphate and sodium borohydride . In the current study, phosphofructokinase was modified by {3H}pyridoxal phosphate and sodium borohydride with or without the addition of citrate to protect the ligand binding site . The modified proteins were digested with trypsin, and the peptides were separated by high-pressure liquid chromatography . A comparison of the tryptic chromatographic profiles showed that while the label was broadly distributed among nine peaks in the elution profile of the enzyme modified in the presence of the protective ligand, a single peptide contained 70% of the total radioactivity of the enzyme modified in the absence of citrate . This peptide was presumed to contain at least part of the citrate inhibitory site of the enzyme . The sequence of the peptide was determined and shown to match with positions 528-536 of phosphofructokinase with the modified residue being Lys-529 . A comparison of the sequence with that of procaryotic phosphofructokinase indicated that a homologous residue in the enzyme from Bacillus stearothermophilis is critical to an allosteric site . A second peptide that was the most abundant labeled peptide in the digest of the enzyme modified in the presence of citrate was found to be identical with the second most abundant peptide of the digest from the unprotected enzyme . This peptide corresponded to residues 681-692 with the lysine at position 684 being the site of phosphopyridoxylation.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Biochem, 1987 Jun 15, 165(3), 547 - 52
DNA-binding properties and primary structure of HB protein from Bacillus globigii; Imber R et al.; The binding of Bacillus globigii HB protein to synthetic deoxyoligonucleotides of different length and sequence has been studied by polyacrylamide gel electrophoresis . Without detectable sequence specificity the protein binds to single-stranded and double-stranded DNA . Under the conditions employed, binding of HB protein to deoxyoligonucleotides with six or less nucleotides per strand cannot be detected while eight or more nucleotide units per strand of single-stranded DNA or base pairs of double-stranded DNA are sufficient for binding . The complete amino acid sequence of HB protein has been determined by manual Edman degradation of tryptic peptides . Like most DNA-binding proteins of its class, HB protein does not contain cysteine, tyrosine or tryptophan residues . The primary structure of HB protein shows 84% homology with the sequence of the related DNA-binding protein II from Bacillus stearothermophilus.

Biochem J, 1987 Jun 15, 244(3), 585 - 90
Purification and some properties of a novel heat-stable cis-toluene dihydrodiol dehydrogenase; Simpson HD et al.; cis-Toluene dihydrodiol dehydrogenase was purified 200-fold from cells of a thermotolerant Bacillus species grown with toluene as the sole source of carbon and energy . The purified enzyme preparation was remarkably heat-stable and exhibited a half-life of 100 min at 80 degrees C, the temperature optimum . The activation energy of the reaction was 36 kJ.mol-1 . Isoelectric focusing indicated that the pI of the native enzyme was 6.4 and that of the denatured enzyme 6.5 . Although the pH optimum was 9.8, the enzyme was most stable at pH 8 . The Mr of the enzyme was approx . 172,000 as determined by gel filtration and 166,000 by polyacrylamide-gel electrophoresis . The enzyme was composed of six apparently identical subunits with Mr values of 29,500 . Kinetic analysis revealed that the Km for cis-toluene dihydrodiol was 92 microM and for NAD+ was 80 microM . The apparent Km values for cis-benzene dihydrodiol and cis-naphthalene dihydrodiol were 330 microM and 51 microM respectively . The enzyme was inhibited by mercurials but was unaffected by metal-ion chelators . Steady-state kinetics and product-inhibition patterns suggested that the enzyme mechanism was ordered Bi Bi.

Eur J Biochem, 1987 Jun 15, 165(3), 581 - 6
Amino acid sequence of the L-lactate dehydrogenase of Bacillus caldotenax deduced from the nucleotide sequence of the cloned gene; Barstow DA et al.; The Bacillus caldotenax L-lactate dehydrogenase gene (lct) has been cloned into Escherichia coli, using the Bacillus stearothermophilus lct gene as a hybridisation probe, and its complete nucleotide sequence determined . The lct structural gene consists of an open reading frame of 951 base pairs commencing with an ATG start codon and followed by a TAA stop codon . Upstream of the gene are putative transcriptional promoter -35 and -10 regions; a ribosome binding site with a predicted delta G of -66.9 kJ/mol is also present six base pairs upstream of the ATG start codon . The B . caldotenax lct gene is highly homologous to the B . stearothermophilus lct gene displaying a DNA sequence homology of 89.7% . Examination of the DNA sequence 3' of the lct gene revealed the presence of two further open reading frames . This suggests that the lct gene may be the first gene of an operon . The deduced amino acid sequence of the L-lactate dehydrogenase (LDH) from B . caldotenax predicted a protein of 317 amino acid residues; comparison with the B . stearothermophilus enzyme revealed only 30 amino acid differences between the two enzymes; thus the enzymes are 90.4% homologous . These amino acid differences must account for the different thermostabilities of the two enzymes . The B . caldotenax lct gene was efficiently expressed in E . coli and the original lct-containing plasmid construct isolated (pKD1) induced the synthesis of LDH at a level of 4.5% of the E . coli soluble cell protein whilst a SmaI subfragment of this clone, (pKD2) produced LDH at a level of 6.9% of the E . coli soluble cell protein . LDH isolated from E . coli cells had the same thermal stability properties as LDH isolated from B . caldotenax cells.

Eur J Biochem, 1987 Jun 15, 165(3), 565 - 70
Isolation and some properties of the site-specific endonuclease and methylase Bme2161 from Bacillus megaterium 216; Matvienko NI et al.; The site-specific endonuclease Bme2161 was isolated as a homogeneous preparation by chromatography on phosphocellulose, hydroxyapatite and heparin-agarose . The molecular mass of the enzyme, determined by gel filtration and by electrophoresis under denaturing conditions, was found to be 60 kDa and 30 kDa respectively . These data indicate that the native enzyme consists of two identical subunits . The enzyme recognized the decreases pentanucleotide sequence 5'-GGACC-3' X 3'-CCTGG-5' and cleaves the sequence as indicated by arrows . The increases optimal concentration for endonuclease reaction is 6-7 mM Mg2+ . The endonuclease relaxes its specificity in the presence of glycerol or dimethyl sulfoxide at low Mg2+ concentration (1-3 mM) . Methylase Bme2161, which protects DNA against endonuclease Bme2161 action by DNA methylation, was isolated from the same bacterial strain.

Nature, 1987 Jun 11-17, 327(6122), 532 - 5
Introduction of foreign DNA into mycobacteria using a shuttle phasmid; Jacobs WR Jr et al.; Mycobacteria are major pathogens of man and animals . There are approximately 10 million cases of tuberculosis world wide with an annual mortality of three million people . Leprosy, caused by Mycobacterium leprae, afflicts over ten million people, primarily in developing countries . M . tuberculosis and mycobacteria of the M . avium-intracellulare-scrofulaceum (MAIS) group are major opportunistic pathogens of patients with acquired immune deficiency syndrome (AIDS) . M . paratuberculosis is the cause of Johne's disease in cattle . Yet, BCG (Bacille Calmette-Guerin), an avirulent strain of M . bovis, is the most widely used human vaccine in the world, having been administered to about 2.5 X 10(9) people since 1948 (ref . 4) . BCG was highly protective against tuberculosis in England, but has been found not to be effective in preventing pulmonary tuberculosis in adults in Southern India . We have initiated studies to develop the methodology for efficient gene transfer in mycobacteria . We have constructed recombinant shuttle phasmids which are chimaeras containing mycobacteriophage DNA into which an E . coli cosmid is inserted . They can replicate in E . coli as plasmids and in mycobacteria as phages, and transfer DNA across both genera . These shuttle vectors permit for the first time the introduction of foreign DNA by infection into M . smegmatis and BCG . By introducing and ultimately expressing genes for protective antigens for a variety of pathogens, it may be possible to develop cultivatable mycobacteria into useful multivaccine vehicles.

J Mol Biol, 1987 Jun 5, 195(3), 755 - 7
Preliminary X-ray diffraction analysis of crystals of Bacillus thuringiensis toxin, a cell membrane disrupting protein; McPherson A et al.; Crystals suitable for high resolution X-ray diffraction analysis have been reproducibly grown of the 24,000 Mr protein insect toxin from Bacillus thuringiensis . This protein, which demonstrates substantial insecticidal activity by inserting into phospholipid membranes, crystallizes as long square needles from polyethylene glycol 4000 at neutral pH . The crystals are of space group P4(1) and have cell dimensions of a = b = 33 A and c = 235 A, which suggests to us a predominantly helical motif for the protein's structure.

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