Bacillus

Nahrung, 1989, 33(9), 839 - 44
Survival and growth of Bacillus cereus in Egyptian bread and its effect on bread staling; Rizk IR et al.; Bread was made from dough previously inoculated with spores of three Bacillus cereus strains at different inoculation levels . Counts of survivors and bread staling were determined in bread during storage at 25 degrees C . The total aerobic bacteria, sporeformer and B . cereus counts in seven different wheat flours were 3.0 x 10(6) to 9.1 x 10(7), 3.2 x 10(5) to 1.5 x 10(6) and 50-100 colony forming units (CFU/g) . About one in a thousand bacteria were found to survive baking . On the other hand, no multiplication took place, however, until the bread had been stored for more than 3 days . Also, survival and multiplication of B . cereus spores was shown to depend on the kind of strains and type of bread . The addition of B . cereus strains to the dough gave bread with higher freshness than the control sample . This was quite noticeable in inoculated pan bread during storage . It is possible to conclude from the results that B . cereus survive the baking process particularly if original contamination is high . Also increasing the freshness of the bread does not necessarily mean better ingredient and processing conditions.

Microbios, 1989, 60(242), 45 - 51
Membrane-bound succinate dehydrogenase of Bacillus pumilus strain 5: effects of modulators of monoelectron transfer; Eze MO et al.; The membrane-bound succinate dehydrogenase (SDH; EC 1.3.99.1) of Bacillus pumilus strain 5 was investigated as succinate:ferricyanide oxidoreductase activity at 27 degrees C . A Km of 8.3 x 10(-3) M was obtained, and the Vmax was 1.8 x 10(-6) mole succinate dehydrogenated min-1 mg-1 membrane protein, at a substrate (succinate) concentration below 40 x 10(-3) M . Above this succinate concentration the Km was 102 x 10(-3) M and the Vmax was 3.7 x 10(-6) mole succinate min-1 mg-1 membrane protein . Para-benzoquinone or 2,4-dinitrophenylhydrazine, in micromolar amounts inhibited the enzyme by serving as an electron sink . Hydroxyl radical (OH.) scavengers, mannitol and benzoate, activated the enzyme, while superoxide dismutase (SOD) had no effect on the enzyme . Thus, the mechanism of electron transfer from succinate to Fe(CN)3-(6) through SDH does not involve superoxide (O2-) as a rate-limiting intermediate.

Cytobios, 1989, 58(233), 85 - 91
Ultrastructural effects of macrotetrolides of Streptomyces griseus LKS-1 in tissues of Culex pipiens larvae; Zizka Z et al.; The isolate of macrotetrolides produced by Streptomyces griseus strain LKS-1 was tested in its effect on the ultrastructure of larvae of Culex pipiens autogenicus . Changes were mainly in mitochondria where the cristae were destroyed and the outer membrane inflated . The endoplasmic reticulum was vacuolized and subsequently the nuclear membranes were seriously affected . Microvilli of the midgut epithelial cells and the surface membrane of these cells were unaltered and there were no changes in the arrangement of cells in the tissues . The effect of macrotetrolides on insect tissues is analogous to the effect of secondary metabolites of fungi such as beauvericin, destruxin E, cyclosporin or tolypin, and differs from the effects of bacterial endotoxins of Bacillus thuringiensis or B . sphaericus.

Prog Clin Biol Res, 1989, 310, 237 - 52
Intravesical therapy comparing BCG, adriamycin, and thiotepa in 200 patients with superficial bladder cancer: a randomized prospective study; Martinez-Pineiro JA et al.; This report presents the second interim analysis of data from a randomized prospective trial that compares the prophylactic effect of 15 intravesical instillations of 50 mg Doxorubicin (ADM), 50 mg of Thiotepa (TTPA), or 150 mg of Bacillus Calmette-Guerin (BCG) against recurrence and progression of superficial transitional cell bladder cancer . Of 202 enrolled patients, 176 patients are currently evaluable after a mean follow-up of 3 years (range, 3-97 months) . The number of patients with recurrence was significantly lower in the BCG group (9/67) than in the ADM group (23/53, p = 0.002) or the TTPA group (20/56, p = 0.003) . The overall recurrence index per 100 patient-months was also significantly lower for the BCG group (BCG vs . ADM, p = 0.07; BCG vs . TTPA, p = 0.001; TTPA vs . ADM, not significant) . BCG was superior in preventing recurrence and progression of high risk tumors (T1, G2-3, multiple growth, and tumors associated with carcinoma in situ) . The recurrence in this group of high risk tumors was for ADM treated patients 12/17, for TTPA treated patients 10/17 and for BCG treated patients 5/23 (BCG vs . ADM, p = 0.002; BCG vs . TTPA, p = 0.016; TTPA vs . ADM, not significant) . Toxicity of intravesical BCG was higher than that of the other drugs, but not limiting the treatment . Bladder irritability occurred in 42% of the patients, granulomatous cystitis in 16.4%, and bladder contraction in 1.5% of the patients . Two patients of the ADM group (2/53 = 3.8%) underwent radical cystectomy for local urothelial progression . One patient (1.9%) in the same group died of distant metastases . The preliminary results suggest that BCG is significantly superior to the chemotherapeutic agents ADM and TTPA when used as an adjuvant intravesical therapy in superficial bladder cancer.

Microbiol Immunol, 1989, 33(7), 527 - 38
Characterization and deposition of the proteins in the outermost layer of Bacillus megaterium spore; Takubo Y et al.; It was proved that three spore coat proteins of 48, 36, and 22 kDa (P48, P36, and P22) were the components of the outermost layer (OL) of Bacillus megaterium ATCC 12872 spore by analysis of the isolated OL . And it was indicated that these proteins were deposited not by disulfide bond, but by ionic and/or hydrophobic bonds on the spore . Among them, P36 and P22 were expected to be located on the very surface of the spore by immunological analysis . In the OL deficient mutant of B . megaterium ATCC 12872, MAE05, whose spore was lacking in these OL proteins and galactosamine-6-phosphate polymer, both P36 and P22 were present in the mother cell cytoplasm and deposited on the forespores, but they disappeared with the lysis of mother cells . An OL protein-releasing factor having proteolytic activity was detected in the culture supernatant at the late sporulating stage of both the wild-type and the mutant strains . But the factor could not act on the proteins of the mature spores and the forespores at t10 (tn indicates n hr after the end of exponential growth) of the wild-type strain . Moreover, P36 and P22 were found in the spores of a revertant of MAE05 which could form galactosamine-6-phosphate polymer, suggesting that this sugar polymer played the role in protecting the OL proteins against the protease-like substance after the deposition.

Acta Leprol, 1989, 7 Suppl 1, 51 - 4
Comparative studies of antigenic glycolipids of mycobacteria related to the leprosy bacillus; Minnikin DE et al.; The leprosy bacillus, Mycobacterium leprae, is a member of a small group of mycobacteria comprising the species Mycobacterium bovis, Mycobacterium marinum, Mycobacterium kansasii, Mycobacterium tuberculosis, Mycobacterium ulcerans and related taxa . This relationship is based on the similarity of the characteristic lipid types in the cell envelope . Mycobacterium leprae produces a phenolic glycolipid antigen which is species specific . This communication reports a comparison of the specificity of the lipid antigens of other members of this group of mycobacteria . Mycobacterium kansasii, in accordance with previous studies, produces phenolic glycolipid and trehalose-based lipooligosaccharide antigens which do not cross react with antisera raised against other mycobacteria . The phenolic glycolipid and an uncharacterised polar glycolipid, with the properties of a lipooligosaccharide, from Mycobacterium marinum are also shown to be specific antigens . An acylated trehalose glycolipid antigen from Mycobacterium tuberculosis H37Rv reacts strongly with antisera raised against the same strain and sera from eight out of ten tuberculosis patients . The phenolic glycolipid antigen, isolated only from Mycobacterium tuberculosis "Canetti" variants, did not react with antisera raised against the type strain, Mycobacterium tuberculosis H37Rv, although it had been shown previously to react with sera from tuberculosis patients . It is apparent that there are populations of the tubercle bacillus which differ in the lipid antigens expressed on their cell surface.

Zentralbl Mikrobiol, 1989, 144(2), 105 - 9
Indolinone derivatives as potential antimicrobial agents; Singh SP et al.; 1-substituted aminomethyl-3-cyclohexylthiosemicarbazone-2-indolinones (I) were tested for their antibacterial activity against Bacillus pumilis, Bacillus brevis and Bacillus megaterium and antifungal activity against Aspergillus flavus, Aspergillus fumigatus and Aspergillus niger . The majority of the compounds were found to exhibit promising antibacterial and antifungal activities . These compounds were also screened for their antiviral action against tobacco mosaic virus in Nicotiana glutinosa plants in vivo as well as in vitro . Most of the compounds had shown significant antiviral activities both in vivo and in vitro.

Bull World Health Organ, 1989, 67(2), 171 - 80
Reduction of enteric infectious disease in rural China by providing deep-well tap water; Wang ZS et al.; Enteric infectious disease (EID), defined here as bacillary dysentery, viral hepatitis A, El Tor cholera, or acute watery diarrhoea, is an important public health problem in most developing countries . This study assessed the impact on EID of providing deep-well tap water (DWTW) through household taps in rural China . For this purpose, we compared the incidence of EID in six study villages (population, 10,290) in Qidong County that had DWTW with that in six control villages (population 9397) that had only surface water . Both the bacterial counts and chemical properties of the DWTW met established hygiene standards for drinking water . The incidence of EID in the study region was 38.6% lower than in the control region; however, the introduction of DWTW supplies did not significantly affect the incidence of bacillary dysentery . These results indicate that the construction and use of DWTW systems with household taps is associated with decreased incidences of El Tor cholera, viral hepatitis A, and acute watery diarrhoea . Since high construction costs have led many authorities to question the value of DWTW, we carried out a cost-benefit analysis of the programme . The cost of constructing a DWTW system averaged US $36,000 at 1983 prices, or US $10.50 per capita . The combined capital and operating costs of a DWTW system were US $1.46 per capita per annum over its 20-year estimated life . The benefits derived from reductions in cost of illness and savings in time to fetch water were 2.2 times the costs at present values Capital outlays were recouped in a 3.6-year payback period and the provision of DWTW proved highly beneficial in both economic and social terms.

Izv Akad Nauk SSSR Biol, 1989 Jan-Feb, (1), 88 - 94
{Isolation, purification and identification of protective substances from a culture broth of germinating Bacillus cereus spores}; Pronin SV et al.; A fraction increasing the resistance of resting spores to UV-irradiation and high temperature has been isolated from the culture medium at the stage of B . cereus st . 96 spore initiation . Amino acid analysis, gas chromatography, electrophoresis, and TLC of the products of acidic and alkaline hydrolysis of the isolated fraction demonstrated that the active component of the fraction was the lipoteichoic acid.

FEMS Microbiol Lett, 1989 Jan 1, 48(1), 13 - 7
Energy-dependent uptake of urea by Bacillus megaterium; Jahns T et al.; Evidence for the existence of an energy-dependent urea uptake system in Bacillus megaterium DSM 90 was obtained by studying the uptake of 14C-urea . In vivo urea uptake and in vitro urease activity differed significantly with respect to temperature- and pH-dependence, kinetic parameters and response towards metabolic inhibitors . Highest uptake activities were observed during exponential growth, and a rapid decrease in urea uptake occurred when cells entered the stationary growth phase and started to sporulate . Significant differences in the uptake rates were observed during growth with different nitrogen sources, suggesting that the formation of the system is under nitrogen control.

Appl Environ Microbiol, 1989 Jan, 55(1), 212 - 8
In vitro metabolism of 2,2'-diaminopimelic acid from gram-positive and gram-negative bacterial cells by ruminal protozoa and bacteria; Denholm AM et al.; Bacillus megaterium GW1 and Escherichia coli W7-M5 were specifically radiolabeled with 2,2'-diamino{G-3H}pimelic acid {( 3H}DAP) as models of gram-positive and gram-negative bacteria, respectively . These radiolabeled bacterial mutants were incubated alone (control) and with mixed ruminal bacteria or protozoa, and the metabolic processes, rates, and patterns of radiolabeled products released from them were studied . Control incubations revealed an inherent difference between the two substrates; gram-positive supernatants consistently contained 5% radioactivity, whereas even at 0 h, those from the gram-negative mutant released 22% . Incubations with ruminal microorganisms showed that the two mutants were metabolized differently and that protozoa were the major effectors of their metabolism . Protozoa exhibited differential rates of engulfment (150 B . megaterium GW1 and 4,290 E . coli W7-M5 organisms per protozoan per h), and they extensively degraded {3H}DAP-labeled B . megaterium GW1 at rates up to nine times greater than those of ruminal bacteria . By contrast, {3H}DAP-labeled E . coli W7-M5 degradation by either ruminal bacteria or ruminal protozoa was more limited . These fundamental differences in the metabolism of the two mutants, especially by ruminal protozoa, were reflected in the patterns and rates of radiolabeled metabolites produced; many were rapidly released from {3H}DAP-labeled B . megaterium GW1, whereas few were slowly released from {3H}DAP-labeled E . coli W7-M5 . Most radiolabeled products derived from {3H}DAP-labeled B . megaterium GW1 were peptides of bacterial peptidoglycan origin . The ruminal metabolism of DAP-containing gram-positive and gram-negative bacteria, even with the same peptidoglycan chemotype, is thus likely to be profoundly different.(ABSTRACT TRUNCATED AT 250 WORDS)

Br J Ophthalmol, 1989 Jan, 73(1), 25 - 8
Bacillus cereus panophthalmitis associated with intraocular gas bubble; al-Hemidan A et al.; It has become increasingly apparent that Bacillus cereus can cause a severe and devastating form of endophthalmitis following penetrating trauma by a metallic object . B . cereus is an uncommon aetiological agent in non-clostridial gas-forming infections . The patient studied in this single case report showed evidence of intraocular gas mimicking gas gangrene infection . The physiology of non-clostridial bacteria producing gas from anaerobic metabolic conditions is reviewed . Further intraocular and systemic complications which may be avoided by accurate and early diagnosis and the use of recommended treatment with antibiotics such as clindamycin.

J Pharm Biomed Anal, 1989, 7(7), 859 - 64
Flow microcalorimetric assay of antibiotics--II . Neomycin sulphate and its combinations with polymyxin B sulphate and zinc bacitracin on interaction with Bacillus pumilus (NCTC 8241); Joslin Kjeldsen N et al.; A flow microcalorimetric assay for Neomycin has been developed which is monitored through interaction of the antibiotic with Bacillus pumilus as the test organism . The assay has better reproducibility (relative standard deviation 2.3%) and is more sensitive than conventional microbiological bioassay (0.5-2 micrograms ml-1) . The effects of combinations with zinc bacitracin, with polymyxin B sulphate, and with both zinc bacitracin and polymyxin B sulphate (both in equimolar proportions), and in those proportions present in the commercial preparation TrisepR (ICI, Macclesfield, UK) have also been investigated . Synergy was observed for the combinations of Neomycin with the other two antibiotics in binary mixtures at the relative proportions found in TrisepR . The addition of all three antibiotics at the levels used in TrisepR did not show synergy . However, addition of all three antibiotics at equimolar concentrations did show synergy . It is suggested that microcalorimetry may be useful in in vitro experiments for exploring the relative proportions required for maximal effect in antibiotic combinations.

Medicina (B Aires), 1989, 49(4), 357 - 9
{Bacteroides distasonis meningitis}; Santoianni JE et al.; A 50 year old woman while undergoing severe treatment for rheumatoid arthritis, developed anaerobic meningitis . The cerebrospinal fluid (CSF) sample was transported and cultivated aerobically and anaerobically . After 48 h at 37 degrees C the anaerobically incubated plate, the enriched fluid thioglycollate medium and the anaerobic culture medium yielded luxuriant growth of an anaerobic Gram negative bacillum . The biochemical and antimicrobial susceptibility patterns were consistent with those for Bacteroides distasonis . Most of the strains of the 5 species included in the Bacteroides fragilis group (B . fragilis, B . vulgatus, B . ovatus, B . thetaiotaomicron and B . distasonis) are resistant to penicillins, cephalosporins of first generation and aminoglycosides . Anaerobic polyresistant flora from an intraabdominal focus (chronic cholecystitis) might have been selected by treatment with gentamicin and cephalotin, and proliferated into meningeal dissemination . It is important that CSF from immunocompromised patients with acute or chronic pulmonary, intraabdominal or cranium-facial infectious processes be transported and cultured in aerobic and anaerobic conditions . These patients must be treated with an initial therapeutic scheme that includes an effective antibiotic for the anaerobic microorganism that may be involved.

Yi Chuan Xue Bao, 1989, 16(5), 389 - 98
{New cloning vectors constructed in Bacillus}; Qiao M et al.; Two new plasmids pNQ216 (4.1kp) and pNQ402 (2.8 kb) were constructed by combining the replication origin of the plasmid pNK289, a cryptic plasmid resides in B . pumilus 289, and the cat-86 gene from plasmid pPL601 . These two plasmids can be maintained steadily in both B . subtilis and B . pumilus . The penetrance of these plasmids on LB medium with Cm (20 micrograms/ml) is 30% higher than that of pPL600 . Thus both of these plasmids can be used as new cloning vectors in Bacillus.

Ann Ig, 1989 Jan-Apr, 1(1-2), 267 - 93
{Trends in tuberculosis in the Ferrara region 1984-1987}; Gregorio P et al.; The Authors investigate some epidemiologic aspects of the patients affected by Tuberculosis in Ferrara's area, during 1984-1987 . The patients with Tuberculosis and submitted to this study were singled out using documents from: Chair and Department of Respiratory Diseases, University of Ferrara School of Medicine, Tresigallo Hospital; Pneumology and Occupational Medicine Service, USL 31, Ferrara; TBC Assistance Bureau, INPS, Ferrara . A total of 563 "cases" of Tuberculosis were recognized; of them, 138 had a relapse of a previous disease, 27 had multiple and contemporary diseases, and 18 had different types of Tuberculosis in various periods of the study . The analysis of the data shows that Tuberculosis is still present in our area, in some cases with clinical features of aggressiveness . In relation to sex and age, a significant increase of Tuberculosis in females during 1986-87, and a significant amount of young patients, were observed . Moreover, a significant rise of extrapulmonary forms, particularly in females, was stressed as well as the onset of primary Tuberculosis during 1987 . The very important problem of bacillar spread, and its implication with a report of Tuberculosis in food-stuff sellers and health-staff, are finally discussed . The Authors' conclusion's the following: a more active epidemiologic surveillance of Tuberculosis is still required.

Appl Biochem Biotechnol, 1989 Jan-Aug, 20-21, 421 - 36
On the mechanism of growth of cells (Bacillus amyloliquefaciens) in the mixed aqueous two-phase system; Alam S et al.; The growth of Bacillus amyloliquefaciens in the aqueous two-phase system, made up of polyethylene glycol, dextran, and water, was investigated . Generally, Bacillus partitions in the dextran phase, but the magnitude of the separation depends largely on the overall composition of polymers in the phase system . The kinetics of growth of Bacillus amyloliquefaciens was studied in the polyethylene glycol-rich continuous phase, dextran-rich dispersed phase, and in the mixed phase . From the kinetic data it appears that increasing the overall polymer composition causes the cells to adsorp at the interface . On the other hand, partition measurements indicate that increasing polymer concentrations make the cell partitioning more one-sided . This anomaly is explained by studying the interfacial adsorption of cells via dynamic surface tension measurements.

J Basic Microbiol, 1989, 29(1), 55 - 60
Functional half-life of the alpha-amylase mRNA of Bacillus licheniformis; Tonkova A et al.; Some aspects of the regulation of alpha-amylase synthesis in Bacillus licheniformis CCM 2205 were investigated . The effect of actinomycin D and chloramphenicol was studied at the level of RNA transcription and translation . alpha-amylase synthesis in Bacillus licheniformis CCM 2205 was practically not altered during the first 20 min after the addition of actinomycin D, although RNA synthesis was almost completely blocked . In contrast to RNA polymerase inhibitor, chloramphenicol stopped immediately the synthesis of alpha-amylase . By using the least squares method the mean half-life of alpha-amylase mRNA was calculated to range from 7.5 to 8.4 min . the mean half-life of cell protein mRNA was determined to range from 2.6 to 3.8 min . Having in mind the immediate effect of chloramphenicol on the alpha-amylase synthesis, it can be concluded that de novo protein synthesis is required in the case of actinomycin D resistant residual synthesis.

Eur J Immunol, 1989 Jan, 19(1), 43 - 7
Efficient mapping and characterization of a T cell epitope by the simultaneous synthesis of multiple peptides; Van der Zee R et al.; Prediction, identification and analysis of T cell epitopes in protein antigens has become a central theme in fundamental and applied immunology . However, while for the characterization of linear B cell epitopes the so-called Pepscan procedure was found to be extremely effective, no such technique has so far been available for T cell studies . Recently, we described the identification and localization of a T cell epitope in a mycobacterial 65-kDa shock protein in the model of adjuvant arthritis . This was done by molecular cloning and conventional solid-phase synthesis techniques . We now show that the delineation of such a T cell epitope and its further characterization can be accomplished in a much more rapid and efficient manner by a modification of the existing Pepscan technique . We show for the first time that several hundreds of peptides, simultaneously synthesized in an automated way on activated polyethylene rods, can be easily recovered from these rods in adequate quantities, enabling a systematic analysis of T cell epitopes . Synthesis of sequentially overlapping peptides along the 65-kDa protein revealed that the adjuvant arthritis T cell clones are fully stimulated by peptides that comprise a minimal sequence of seven residues, corresponding to positions 180-186 in the sequence of the 65-kDa protein of M . bovis Bacillus Calmette Guerin (BCG) . Detailed examination of the epitope by peptides containing a single amino acid substitution showed that, apart from one conservative replacement (Glu----Asp), the requirement for the native residue at all positions in peptide 180-186 was absolute for full T cell stimulation . Their indispensability was confirmed with deletion and insertion peptides . It is concluded that the occurrence of indifferent or spacer residues in a minimal stimulatory sequence, as observed by others, is not a general feature of T cell epitopes.

J Bacteriol, 1989 Jan, 171(1), 375 - 82
A single gene directs synthesis of a precursor protein with beta- and alpha-amylase activities in Bacillus polymyxa; Uozumi N et al.; The Bacillus polymyxa amylase gene comprises 3,588 nucleotides . The mature amylase comprises 1,161 amino acids with a molecular weight of 127,314 . The gene appeared to be divided into two portions by the direct-repeat sequence located at almost the middle of the gene . The 5' region upstream of the direct-repeat sequence was shown to be responsible for the synthesis of beta-amylase . The 3' region downstream of the direct-repeat sequence contained four sequences homologous with those in other alpha-amylases, such as Taka-amylase A . The 48-kilodalton (kDa) amylase isolated from B . polymyxa was proven to have alpha-amylase activity . The amino acid sequences of the peptides generated from the 48-kDa amylase showed complete agreement with the predicted amino acid sequence of the C-terminal portion . The B . polymyxa amylase gene was therefore concluded to contain in-phase beta- and alpha-amylase-coding sequences in the 5' and 3' regions, respectively . A precursor protein, a 130-kDa amylase, directed by a plasmid, pYN520, carrying the entire amylase gene, had both beta- and alpha-amylase activities . This represents the first report of a single protein precursor in procaryotes that gives rise to two enzymes.

Int J Food Microbiol, 1988 Dec 31, 7(4), 349 - 52
The inhibition of the growth of Bacillus cereus in liver sausage; Asplund K et al.; The growth of Bacillus cereus is a problem in liver sausage especially when the sausages are stored at high temperatures . Even concentrations of greater than 10(6)/g have been detected . In this study we found that when combining glucono-delta-lactone, sodium erythorbate and citric acid with sodium nitrite and salt the growth of B . cereus could be delayed or totally inhibited.

J Mol Biol, 1988 Dec 20, 204(4), 1041 - 3
Crystallization of a ternary complex of lactate dehydrogenase from Bacillus stearothermophilus; Wigley DB et al.; Bacillus stearothermophilus lactate dehydrogenase was purified from an overexpressing Escherichia coli cell line . The enzyme has been crystallized in several different forms . All of these crystal forms were grown in the presence of NADH, sodium oxamate and fructose 1,6-bisphosphate . Three crystal forms have been characterized, an orthorhombic P2(1)2(1)2 (type III, a = 86 A, b = 105 A, c = 136 A) and two monoclinic P21 forms (type IV, a = 85 A, b = 118 A, c = 136 A, beta = 96 degrees; type V, a = 112 A, b = 85 A, c = 136 A, beta = 91 degrees) . Precession photographs from these crystal forms are very alike, suggesting the molecular packing to be similar in all three forms . The P21 type IV crystals diffract to beyond 2 A spacing and are stable to irradiation with X-rays . A complete medium-resolution (4.7 A) dataset has been collected from a single crystal using synchrotron radiation . Rotation function studies with these data show the two tetramers of the asymmetric unit to be in very similar orientations . Higher-resolution data are being collected.

J Mol Biol, 1988 Dec 20, 204(4), 973 - 94
Crystal structure of the complex of phosphofructokinase from Escherichia coli with its reaction products; Shirakihara Y et al.; The crystal structure of Escherichia coli phosphofructokinase complexed with its reaction products fructose 1,6-bisphosphate (Fru1,6P) and ADP/Mg2+, and the allosteric activator ADP/Mg2+, has been determined at 2.4 A resolution . The structure was solved by molecular replacement using the known structure of Bacillus stearothermophilus phosphofructokinase, and has been refined to a crystallographic R-factor of 0.165 for all data . The crystallization mixture contained the substrate fructose 6-phosphate, but the electron density maps showed clearly the presence of the product fructose 1,6-bisphosphate, presumably formed by the enzyme reaction with contaminating ATP . The crystal consists of tetrameric molecules with subunits in two different conformations despite their chemical identity . The magnesium ion in the "closed" subunit bridges the phosphate groups of the two products . In the "open" subunit, the products are about 1.5 A further apart, with the Mg2+ bound only to ADP . These two conformations probably represent two successive stages along the reaction pathway, in which the closure of the subunit is required to bring the substrates sufficiently close to react . This conformational change within the subunit is distinct from the quaternary structure change seen previously in the inactive T-state conformation . It is probably not involved in the co-operativity or allosteric control of the enzyme, since the co-operative product fructose 1,6-bisphosphate is not moved, nor are the subunit interfaces changed . The structure of the enzyme is similar to that of B . stearothermophilus phosphofructokinase, and confirms the location of the sites for the two reaction products (or substrates), and of the effector site binding the activator ADP/Mg2+ . However, this structure gives a clearer picture of the active site, and of the interactions between the enzyme and its reaction products.

Science, 1988 Dec 16, 242(4885), 1541 - 4
A specific, highly active malate dehydrogenase by redesign of a lactate dehydrogenase framework; Wilks HM et al.; Three variations to the structure of the nicotinamide adenine dinucleotide (NAD)-dependent L-lactate dehydrogenase from Bacillus stearothermophilus were made to try to change the substrate specificity from lactate to malate: Asp197----Asn, Thr246----Gly, and Gln102----Arg) . Each modification shifts the specificity from lactate to malate, although only the last (Gln102----Arg) provides an effective and highly specific catalyst for the new substrate . This synthetic enzyme has a ratio of catalytic rate (kcat) to Michaelis constant (Km) for oxaloacetate of 4.2 x 10(6)M-1 s-1, equal to that of native lactate dehydrogenase for its natural substrate, pyruvate, and a maximum velocity (250 s-1), which is double that reported for a natural malate dehydrogenase from B . stearothermophilus.

Biochim Biophys Acta, 1988 Dec 16, 963(3), 423 - 8
Chemical cross-linking and its effect on fatty acid synthetase activity in intact chloroplasts from Euglena gracilis; Worsham LM et al.; Intact chloroplasts were isolated from Euglena gracilis variety bacillaris, aliquots were exposed to several different chemical cross-linking reagents . The reagents penetrated the triple membrane of Euglena chloroplasts . This was shown by gradient acrylamide gel electrophoresis under denaturing conditions . The activity of the nonaggregated fatty acid synthetase of Euglena was located within the chloroplast stroma, and the effects of dimethylsuberimidate cross-linking on the activity of the enzyme system were examined . The acyl-carrier protein concentration in the chloroplast was measured at about 0.24 mM.

Gene, 1988 Dec 15, 73(1), 209 - 14
Chloramphenicol acetyltransferase specified by cat-86: relationship between the gene and the protein; Laredo J et al.; Gene cat-86 is chloramphenicol (Cm)-inducible and specifies Cm acetyltransferase, CAT-86 . The gene was previously cloned from the DNA of a strain of Bacillus pumilus . In the present study we report the construction of a constitutively expressed version of cat-86 that permits high-level expression of the gene on a plasmid in B . subtilis . A method is described that allows very rapid purification of CAT-86 protein to homogeneity . The sequence of 13 N-terminal amino acids of purified CAT-86, as well as the 26.6-kDa size of the subunit protein, agree with predictions made based on the nucleotide sequence of the gene . The Mr of the native enzyme suggests that CAT-86 is a trimer consisting of three identical protein subunits . Our studies demonstrate that cat-86 provides a convenient system for analyzing relationships between a gene and a multimeric enzyme in the B . subtilis background.

Biochemistry, 1988 Dec 13, 27(25), 9056 - 62
Gene cloning and sequence determination of leucine dehydrogenase from Bacillus stearothermophilus and structural comparison with other NAD(P)+-dependent dehydrogenases; Nagata S et al.; The gene for leucine dehydrogenase (EC 1.4.1.9) from Bacillus stearothermophilus was cloned and expressed in Escherichia coli . The selection for the cloned gene was based upon activity staining of the replica printed E . coli cells . A transformant showing high leucine dehydrogenase activity was found to carry an about 9 kilobase pair plasmid, which contained 4.6 kilobase pairs of B . stearothermophilus DNA . The nucleotide sequence including the 1287 base pair coding region of the leucine dehydrogenase gene was determined by the dideoxy chain termination method . The translated amino acid sequence was confirmed by automated Edman degradation of several peptide fragments produced from the purified enzyme by trypsin digestion . The polypeptide contained 429 amino acid residues corresponding to the subunit (Mr 49,000) of the hexameric enzyme . Comparison of the amino acid sequence of leucine dehydrogenase with those of other pyridine nucleotide dependent oxidoreductases registered in a protein data bank revealed significant sequence similarity, particularly between leucine and glutamate dehydrogenases, in the regions containing the coenzyme binding domain and certain specific residues with catalytic importance.

Asepsis, 1989 1st Quarter, 11(1), 14 - 8
Immunization of health care workers: new concepts . Forum; Keys TE et al.; During the course of their work activities, health care workers often come into contact with patients who have communicable diseases . While barrier precautions and isolation procedures are useful in preventing transmission of nosocomial pathogens, immunizations available for some infectious diseases are a primary line of defense . It is important for health care workers not only to understand the method of transmission of these diseases, but to receive vaccines for those diseases that they may be at risk of acquiring . Each author was asked to discuss the hepatitis B vaccines which are currently available, from the standpoint of acceptability, side effects and cost . They were also questioned about the circumstances under which health care workers should be immunized against influenza, and about Bacille Calmette-Guerin (BCG) vaccine's current role in the prevention of tuberculosis . Finally, we asked the authors to address vaccines of interest to health care workers which are currently in developmental stages, and are likely to become available within the next several years.

Hansenol Int, 1988 Dec, 13(2), 34 - 6
{Current status of surgery in the comprehensive treatment of Hansen's disease}; Virmond M et al.; In the last decades the neural component of Hansen's disease has achieved its place of prime importance among other manifestations of the disease . Provided that there is a close relationship between neural involvement and deformities and that hitherto antileprosy drugs are able only to kill and prevent bacillary growth and not able to interrupt the immunological features of the disease, we can expect a significant load of patients with some degree of disability, including those in regular treatment . Surgery plays an important role in control programmes since it has not just the single aim to restore lost function but also to prevent further damage and to improve patient's self-confidence.

Jpn J Genet, 1988 Dec, 63(6), 537 - 41
Tripeptide, Arg-Gly-Asp, inhibits the transfection of protein-linked DNA of bacteriophage M2; Kobayashi H et al.; The effect of tripeptide, Arg-Gly-Asp (RGD), on the transfection activity of Bacillus phage M2 DNA was examined . The transfection activity decreased when M2 DNA was preincubated with RGD before it was added to competent cells.

J Gen Microbiol, 1988 Dec, 134 ( Pt 12), 3269 - 76
Cloning and nucleotide sequence of senN, a novel 'Bacillus natto' (B . subtilis) gene that regulates expression of extracellular protein genes; Wong SL et al.; A new 'Bacillus natto' gene, senN, that regulates the expression of several extracellular proteins in B . subtilis has been cloned and sequenced . senN codes for a small, highly basic protein with an amino acid sequence different from the products coded by the regulatory genes sacQ, sacV, prtR and hpr . SenN stimulates gene expression at the transcriptional level . A putative homologous locus has been detected in the B . subtilis chromosome by Southern blotting.

J Gen Microbiol, 1988 Dec, 134 ( Pt 12), 3179 - 85
The metabolism of aromatic ring fission products by Bacillus stearothermophilus strain IC3; Adams D et al.; Bacillus stearothermophilus IC3 degraded the meta cleavage product of catechol, 2-hydroxymuconic semialdehyde, to pyruvate and acetaldehyde via the 4-oxalocrotonate pathway . The pathway was identical to those previously delineated in several mesophilic organisms . However, all the enzymes showed activity at 55 degrees C and other properties (substrate specificities and effects of metal ions) also differed from those displayed by the mesophilic enzymes . All enzymes of this meta cleavage pathway, except the 2-hydroxy-6-oxohepta-2,4-dienoate hydrolase and 4-hydroxy-2-oxovalerate aldolase activities, were induced by growth on phenol.

Mol Gen Mikrobiol Virusol, 1988 Dec, (12), 29 - 33
{Variability of Bacillus thuringiensis bacteriophages with C2-morphology}; Kuzin AI et al.; Biological as well as physicochemical properties of Bacillus thuringiensis bacteriophages "17" and "7/13" having C2-morphology and isolated from factory phagolysates were studied . The bacteriophages are identical in the lytic spectrum++, morphology, size, GC-content, have the same buoyant density . The physical map for restriction endonucleases EcoRI, HindIII, SalGI and MvaI has been constructed of the bacteriophages DNA . Heteroduplex analysis has revealed the nonhomologous region of the deletion-insertion type as a 0.8 Md loop . The bacteriophages "17" and "7/13" are concluded to be closely related but not identical.

Gastroenterol Jpn, 1988 Dec, 23(6), 646 - 51
Enzymatic determination of serum 12 alpha-hydroxy bile acid concentration with 12 alpha-hydroxysteroid dehydrogenase; Tamasawa N et al.; A simple colorimetric enzymatic assay for determination of serum 12 alpha-hydroxy bile acids was developed using 12 alpha-hydroxysteroid dehydrogenase (HSD) . The enzymes were extracted from Bacillus sphaericus . The principle of the method is as follows: 12 alpha-hydroxy bile acids are converted to 12-oxo bile acids using 12 alpha-HSD with the conocomitant reduction of NAD to NADH, and then the hydrogen of the generated NADH is transferred by diaphorase to NTB to yield diformazan . Finally, the color of resultant diformazan was measured . The specificity and precision of this assay method were satisfactory . A linear relationship was noted between the amount of 12 alpha-hydroxy bile acids and the degree of absorbance in the range of 6.7 to 215 microM . The fasting values for serum 12 alpha-hydroxy bile acid in 10 patients with liver diseases ranged widely from 7.6 to 91.1 microM, and values obtained with this assay agreed closely with those obtained by gas-liquid chromatography (r = 0.94, p less than 0.001) . The assay is convenient, rapid, and specific for the measurement of 12 alpha-hydroxy bile acid concentrations in the serum of patients with liver diseases.

J Am Vet Med Assoc, 1988 Dec 1, 193(11), 1425 - 8
Clinical and clinicopathologic findings in two foals infected with Bacillus piliformis; Humber KA et al.; Bacillus piliformis infection (Tyzzer's disease) in foals is rarely observed clinically because of the peracute course of the disease . Clinical and clinicopathologic findings as well as information on therapeutic attempts in two foals are described . Clinicopathologic abnormalities common to both cases included leukopenia, hyperfibrinogenemia, metabolic acidosis, and hypoglycemia . Treatment was unsuccessful in both cases.

Chest, 1988 Dec, 94(6), 1296 - 8
Miliary tuberculosis due to intravesical bacillus Calmette-Guerin therapy; Gupta RC et al.; While receiving treatment for bladder carcinoma with intravesical BCG, a 78-year-old man developed a clinical illness and roentgenographic manifestation of miliary tuberculosis . The transbronchial lung biopsy demonstrated granulomas with giant cells . Treatment with antituberculosis therapy resulted in complete resolution of the illness . The pathogenesis of this complication was considered to be due to pulmonary infection by BCG from the bladder source and differs from previously reported cases of interstitial pulmonary infiltrates which more likely represent a hypersensitivity reaction to BCG.

Pediatrics, 1988 Dec, 82(6), 909 - 13
Bacillus species isolates from cerebrospinal fluid in patients without shunts; Feder HM Jr et al.; Of 849 CSF cultures done at Hartford Hospital, nine were positive for nonanthrax Bacillus species . Differentiation of true nonanthrax Bacillus species infection from contamination requires careful consideration of the clinical findings, the clinical course, and the laboratory data . In seven patients the nonanthrax Bacillus species represented contamination . In two patients the nonanthrax Bacillus species represented true infection . In one of these infected patients, nonanthrax Bacillus species complicated a cranial gun shot wound . Bacillus cereus meningitis developed in the second patient, a premature infant, following sepsis from a contaminated IV catheter . Nonanthrax Bacillus species, especially B cereus, can be resistant to penicillins and cephalosporins when nonanthrax Bacillus species infections are being treated, susceptibility testing should always be performed.

J Bacteriol, 1988 Dec, 170(12), 5908 - 12
Role of menaquinone in inactivation and activation of the Bacillus cereus forespore respiratory system; Escamilla JE et al.; The respiratory systems of the Bacillus cereus mother cell, forespore, and dormant and germinated spore were studied . The results indicated that the electron transfer capacity during sporulation, dormancy, and germination is related to the menaquinone levels in the membrane . During the maturation stages of sporulation (stages III to VI), forespore NADH oxidase activity underwent inactivation concomitant with a sevenfold decrease in the content of menaquinone and without major changes in the content of cytochromes and segment transfer activities . During the same period, NADH oxidase and menaquinone levels in the mother cell compartment steadily decreased to about 50% at the end of stage VI . Dormant spore membranes contained high levels of NADH dehydrogenase and cytochromes, but in the presence of NADH, they exhibited very low levels of O2 uptake and cytochrome reduction . Addition of menadione to dormant spore membranes restored NADH-dependent respiration and cytochrome reduction . During early germination, NADH-dependent respiration and cytochrome reduction were restored simultaneously with a fourfold increase in the menaquinone content; during germination, no significant changes in cytochrome levels or segment electron transfer activities of the respiratory system took place.

Infect Immun, 1988 Dec, 56(12), 3196 - 200
H-2-linked control of in vitro gamma interferon production in response to a 32-kilodalton antigen (P32) of Mycobacterium bovis bacillus Calmette-Guérin; Huygen K et al.; A 32-kilodalton protein antigen (P32) was previously purified to homogeneity from culture filtrate of Mycobacterium bovis BCG (J . De Bruyn, K . Huygen, R . Bosmans, M . Fauville, R . Lippens, J . P . Van Vooren, P . Falmagne, H . G . Wiker, M . Harboe, and M . Turneer, Microb . Pathog . 2:351-366, 1987) . Spleen cells from BCG-sensitized mice produce significant amounts of gamma interferon (IFN-gamma) in response to this P32 protein . The amount of secreted IFN-gamma is influenced by mouse genotype, with C57BL/6 (H-2b), C57BL/10 (H-2b), and 129/Sv (H-2b) mice producing about four times more than BALB/c (H-2d), CBF1 (H-2d/b), and DBA/2 (H-2d) mice do . Analysis of seven recombinant inbred strains derived from the BALB/c x C57BL/6 cross and of congenic mice differing in major histocompatibility complex-coding chromosome 17 fragments indicates a probable H-2-linked control of this IFN-gamma induction, with H-2b cells producing high titers and H-2d cells producing low titers in response to the P32 antigen.

Mol Gen Genet, 1988 Dec, 215(1), 181 - 3
Expression of a cloned beta-glucanase gene from Bacillus amyloliquefaciens in an Escherichia coli relA strain after plasmid amplification; Hecker M et al.; Amino acid starvation of cells of the Escherichia coli relA strain, CP79, which cannot accumulate guanosine tetraphosphate (ppGpp) in response to amino acid limitation, increased the pEG1 plasmid content about 5- to 7-fold in comparison with exponentially growing cells (pEG1:pBR322 with an insertion of Bacillus amyloliquefaciens DNA coding for beta-glucanase) . In contrast, no pEG1 amplification occurred in E . coli CP78, the stringently controlled counterpart, after amino acid starvation . In order to verify these results, the plasmid DNA content was monitored by measuring the expression of pEG1-encoded beta-glucanase from B . amyloliquefaciens both before and after plasmid amplification . When amino acid starved CP79 cells were given an additional dose of amino acids, a more than 10-fold increase in pEG1-encoded beta-glucanase activity (per cell mass) was measured . This increase in enzyme activity correlates with pEG1 amplification during amino acid limitation . Under comparable conditions the activity of beta-glucanase was not increased in strain CP78, which did not amplify the plasmid . We suggest that the replication of pEG1 in amino acid starved E . coli cells is somehow under negative control by ppGpp . Moreover, we found the Bacillus beta-glucanase in E . coli relA cells to be excreted into the growth medium after starvation and overexpression.

J Biochem (Tokyo), 1988 Dec, 104(6), 985 - 8
Solubilization and properties of UDP-D-glucose:N-acetylglucosaminyl pyrophosphorylundecaprenol glucosyltransferase from Bacillus coagulans AHU 1366 membranes; Kumita K et al.; The glucosyltransferase which catalyzes the conversion of GlcNAc-PP-undecaprenol into Glc(beta 1----4)GlcNAc-PP-undecaprenol in the presence of UDP-glucose was solubilized from Bacillus coagulans AHU 1366 membranes by treatment with 0.1% Triton X-100 and partially purified by means of column chromatography on Sephacryl S-300 and DEAE-Sephacel . The final preparation was virtually free from other enzymes involved in the de novo synthesis of teichoic acid . The enzyme had a pH optimum of 6.6-8.0 and a Km value for UDP-glucose of 21 microM . The enzyme required 40 mM MgCl2, 0.6 M KCl, and 0.1% Nonidet P-40 for full activity.

Cancer Metastasis Rev, 1988 Dec, 7(4), 289 - 309
Helper strategy in tumor immunology: expansion of helper lymphocytes and utilization of helper lymphokines for experimental and clinical immunotherapy; Forni G et al.; Two main kinds of immune strategy are possible against neoplasia . The first potentiates a selected effector arm . In vitro culture with exogenous interleukin-2 (IL-2) increases the activity of natural killer cells and leads to the expansion of T cytotoxic lymphocytes . Systemic reinfusion of both of these cells with high doses of IL-2 mediates the regression of a variety of murine and human tumors . In an alternative strategy, a few regulatory lymphocytes turn on immune reactivity by triggering a cascade of interconnected effector functions . The efficacy of this strategy rests on the repertoire of effector mechanisms moved to action . An effective immunoregulatory maneuver is the addition of helper determinants on the surface of tumor cells . Its power can be further increased by the pre-induction of helper T lymphocytes specific to the helper determinants . This approach can be achieved in mice by coupling muramyl dipeptides to tumor cells, along with eliciting T lymphocytes specifically reactive to Bacillus Calmette-Guerin . Noncytotoxic T helper lymphocytes produce factors which recruit nonspecific (macrophages) as well as specific (cytolytic T lymphocytes) anti-tumor attacking cells . In this way protection can be afforded against primary tumors and metastases, as well as leukemia cells . As the activity of helper lymphocytes rests mostly on lymphokine release, the use of molecularly defined lymphokines mimicking T-helper functions has also been attempted . In a few experimental models, the association of low doses of IL-2 with non-reactive lymphocytes from tumor-bearing mice promotes an effective anti-tumor reaction in the host . Moreover, the combination of distinct lymphokines can also build a molecularly defined helper system able to activate in sequence non-specific and specific anti-tumor reactions in vivo . Trials intended to evaluate the clinical impact of these helper approaches in the management of human tumors are being started or are already under way.

J Am Mosq Control Assoc, 1988 Dec, 4(4), 470 - 8
An evaluation of Gambusia affinis and Bacillus thuringiensis var . israelensis as mosquito control agents in California wild rice fields; Kramer VL et al.; The mosquito control potential of the mosquitofish and Bacillus thuringiensis var . israelensis (Bti) were evaluated in experimental wild rice fields in Lake County, California . Fields were assigned one of six treatment: control, 1.1 kg/ha G . affinis, 3.4 kg/ha G . affinis, Bti only (6 kg/ha Vectobac granules), 1.1 kg/ha G . affinis plus Bti and 3.4 kg/ha G . affinis plus Bti . Gambusia affinis, at both release rates, significantly reduced the mosquito population at densities exceeding 100 fish/minnow trap . Treatments with Bti significantly reduced larval population; however, the populations in the fields without fish rebounded to pretreatment levels within two weeks . In fields stocked with G . affinis and treated with Bti, populations remained low after Bti treatment . Nontarget populations of arthropods were significantly lower in fields stocked with G . affinis than in fields without fish on one or more sampling dates.

J Am Mosq Control Assoc, 1988 Dec, 4(4), 448 - 52
Efficacy and longevity of Bacillus sphaericus 2362 formulations for control of mosquito larvae in dairy wastewater lagoons; Mulla MS et al.; Bacillus sphaericus strain 2362 was evaluated for the control of Culex larvae in dairy wastewater lagoons . Both initial and long-term efficacy were studied . Two primary powder preparations of ABG-6184 yielded mediocre and short-term control at the rates of 0.25 and 0.5 lb/acre (0.26 and 0.56 kg/ha), while level of control and persistence greatly increased as the dosages were increased to 1.0, 2.0 and 4.0 lb/acre (1.12, 2.24 and 4.48 kg/ha) . The 1.0 and 2.0 lb/acre rates yielded almost 100% control for 4 weeks and the 4.0 lb/acre rate yielded control (99%) for 49 days or longer . A flowable concentrate preparation (BSP-2) yielded complete initial and persistent control of larvae for 14-21 days at 2.0, 4.0 and 5.0 lb/acre (2.24, 4.49 and 5.6 kg/ha) . Granular formulations of B . sphaericus 2362 (ABG-6185) were also evaluated at 2.5, 5.0, 7.5, 10.0 and 20.0 lb/acre (2.8, 5.6, 8.4, 11.2 and 22.4 kg/ha) of the granules . Some of the formulations were more active than the others, yielding excellent initial and persistent control (80% +) for 14-21 days with one treatment.

Biochim Biophys Acta, 1988 Nov 23, 957(2), 178 - 84
Reactivity of the thiol groups of Escherichia coli phosphofructo-1-kinase; Banas T et al.; Modification of Escherichia coli phosphofructo-1-kinase (6-phosphofructokinase; EC 2.7.1.11) with several thiol modifying reagents led to a pseudo-first-order loss of activity that was associated with the modification of a single cysteine residue, identified as the cysteine at position 119 in the protein sequence . This cysteine was protected from reaction with vinyl pyridine, bromopyruvate, and dithionitrobenzoic acid by the substrate, fructose-6-P . In the crystal structure of the highly homologous phosphofructokinase from Bacillus stearothermophilus, cysteine 119 is sufficiently distant from the catalytic site to exclude a direct steric inhibition of the binding of substrate as a mechanism of inactivation for the modification . Thus, the inhibition is unlikely to be a direct one but to be the result of interference with the conformational change that is associated with fructose-6-P binding . A second thiol, position 283, was shown to be protected from reaction when the enzyme was in its native conformation . In contrast to the previously published sequence for the E . coli enzyme six cysteines as opposed to seven have been found both in enzyme from strain LE392 and in enzyme produced by a plasmid that was derived from pLC 16-4 . The position in question, 75, was identified as phenylalanine.

Mikrobiologiia, 1988 Nov-Dec, 57(6), 992 - 5
{Growth and spore formation in Bacillus thuringiensis at high substrate concentrations}; Sakharova ZV et al.; The work was aimed at studying the effect exerted by elevated concentrations of glucose, yeast extract and acetate on the growth of Bacillus thuringiensis subsp . galleriae, strain 69-6, and on the formation of spores and crystals by it . Glucose concentrations from 30 to 100 g per litre did not prevent spore formation . Yeast extract inhibited spore formation to a greater extent and stopped it almost completely at a concentration of 20 g per litre . Acetate at a concentration of 1.0 to 10 g per litre delayed spore formation and produced a less action on crystal formation, so that those processes were uncoupled in time.

Mikrobiologiia, 1988 Nov-Dec, 57(6), 1024 - 30
{Immunochemical characteristics of a lipopolysaccharide from Pseudomonas aurantiaca}; Zdorovenko GM et al.; A lipopolysaccharide was isolated from Pseudomonas aurantiaca IMB 31 by extraction with aqueous phenol and purified by ultracentrifugation . The lipopolysaccharide was confined to the phenol phase . Fucosamine (2-amino-2,6-dideoxygalactose) (36%) and bacillosamine (2,4-diamino-3,4,6-trideoxyglucose) (23%) were identified as hypothetic components of the O-chain in the carbohydrate moiety of the macromolecule using the techniques of paper chromatography, gas-liquid chromatography and ion-exchange chromatography on an amino acid analyser . Rhamnose, glucose, galactose, glucosamine and galactosamine were detected as hypothetical components of the core in the lipopolysaccharide composition, as well as 2-keto-3-deoxyoctonic acid, heptose, alpha-alanine and phosphorus, usual components of the core in Pseudomonas . The following predominant fatty acids were identified in the composition of lipid A using the techniques of gas-liquid chromatography with standard compounds and gas-liquid mass spectrometry: 3-OH C10:0 (14.4%), C12:0 (30.5%), 2-OH C12:0 (14.9%), 3-OH C12:0 (17.4%), C16:0 (9.9%) . The serological relationship between P . aurantiaca strains was studied, and their phylogenetic relationship with P . fluorescens is discussed.

Equine Vet J, 1988 Nov, 20(6), 444 - 7
BCG emulsion immunotherapy of equine sarcoid; Vanselow BA et al.; Of 61 horses with sarcoids treated with intralesional injection of a double emulsion incorporating inactivated bacillus Calmette Guerin organisms, 36 (59 per cent) showed complete regression and 11 (18 per cent) showed partial regression . The majority of cases required only one treatment . Not all sarcoids were responsive to this therapy; those not responding were usually large or on horses with multiple sarcoids.

J Gen Microbiol, 1988 Nov, 134 ( Pt 11), 3001 - 10
Functional relationships between L- and D-alanine, inosine and NH4Cl during germination of spores of Bacillus cereus T; Preston RA et al.; The results of a physiological study of the interaction between NH4Cl, inosine, and the stereoisomers of alanine during germination of spores of Bacillus cereus T are presented . Detailed kinetics for the germination of unheated spores in moderate concentrations of L-alanine (in the absence of auto-inhibition due to alanine racemase) are established, as is the specificity of the stimulatory effect of NH4Cl in relation to other salts, amines, and germinants . The results suggest that NH4Cl and inosine affect an early step in germination closely related to the function of an L-alanine receptor.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1988 Nov, 187(1), 87 - 9
{The contamination of a hospital with Bacillus cereus without an indication of cases of infection}; Nikodemusz I et al.; A hygienic investigation of a hospital revealed B . cereus in 60.5% (66 out of 109) of swab specimens taken from walls, apparatuses, airing devices and bedclothes . These findings were not associated with evidence of human infection caused by the organism within the hospital . In a similar investigation three weeks later, only 22.5% of the environmental specimens were positive for B . cereus . The hospital contamination with B . cereus has thus been virtually temporary and epidemiologically silent . Considering the potential pathogenicity of the organism however, its presence in the hospital environment cannot be approved of.

Mol Gen Genet, 1988 Nov, 214(3), 365 - 72
Isolation and characterization of EG2158, a new strain of Bacillus thuringiensis toxic to coleopteran larvae, and nucleotide sequence of the toxin gene; Donovan WP et al.; A novel strain of Bacillus thuringiensis was isolated from soybean grain dust from Kansas and found to be toxic to larvae of Leptinotarsa decemlineata (Colorado potato beetle) . The strain (EG2158) synthesized two parasporal crystals: a rhomboid crystal composed of a 73116 dalton protein of approximately 30 kDa . Plasmid transfer and gene cloning experiments demonstrated that the 73 kDa protein was encoded on an 88 MDa plasmid and that the protein was toxic to the larvae of Colorado potato beetle (CPB) . The sequence of the 73 kDa protein, as deduced from the sequence of its gene (cryC), was found to have regions of similarity with several B . thuringiensis crystal proteins: the lepidopteran-toxic P1 proteins of var . kurstaki and berliner, the lepidopteran- and dipteran-toxic P2 (or CRYB1) protein of var . kurstaki, and the dipteran-toxic 130 kDa protein of var . israelensis . While B . megaterium cells harboring the cryC gene from EG2158 synthesized significant amounts of the 73 kDa CRYC protein, Escherichia coli cells did not . The cryC-containing B . megaterium cells produced rhomboid crystals that were toxic to CPB larvae.

Mol Microbiol, 1988 Nov, 2(6), 727 - 33
Germination-specific cortex-lytic enzyme is activated during triggering of Bacillus megaterium KM spore germination; Foster SJ et al.; Antibodies were raised against purified germination-specific cortex-lytic enzyme (GSLE) from spores of Bacillus megaterium KM which neutralized the ability of GSLE to germinate permeabilized spores . Western blotting of dormant spore and vegetative cell fractions separated by SDS-PAGE demonstrated that GSLE is spore-specific and that greater than 90% of the GSLE is associated with the dormant spore cortex peptidoglycan as a phosphorylated 63kD pro-form, which could only be visualized after lysozyme digestion of the peptidoglycan . During germination, the 63kD pro-form of GSLE is processed to release the active enzyme, which had an apparent molecular weight of 30kD . Inhibitor studies demonstrated that GSLE activation occurs as part of the commitment reaction and thus represents the first-identified enzymatic event to occur during germination triggering . Proteins that cross-react with anti-GSLE sera are present in spore fractions of other species.

Microbiol Sci, 1988 Nov, 5(11), 333 - 4, 339
Molecular genetic approaches to the pathogenesis of bacillary dysentery; Yoshikawa M et al.; Bacillary dysentery is an invasive infectious disease of the human colon . At least three genetic loci on the chromosome and a huge plasmid have been implicated in its pathogenesis . Results obtained from molecular genetic studies, mainly of the genes, virG, virF and kcpA are discussed.

Prikl Biokhim Mikrobiol, 1988 Nov-Dec, 24(6), 784 - 8
{Use of starch-containing media for cultivation of Bacillus mesentericus}; Kovalenko EA et al.; When cultivating Bacillus mesentericus to produce proteinases it is advisable to use more available and cheap carbon sources--native maize meal or potato starch--instead of maltose; the products of their complete hydrolysis inhibit the biosynthesis of enzymes.

Gene Anal Tech, 1988 Nov-Dec, 5(6), 116 - 24
Cloning of the BamHI methyl transferase gene from Bacillus amyloliquefaciens; Connaughton JF et al.; We wish to report the initial characterization of a recombinant clone containing the BamHI methylase gene . Genomic chromosomal DNA purified from Bacillus amyloliquefaciens was partially cleaved with HindIII, fractionated by size, and cloned into pSP64 . Plasmid DNA from this library was challenged with BamHI endonuclease and transformed into Escherichia coli HB101 . A recombinant plasmid pBamM6.5 and a subclone pBamM2.5 were shown to contain the BamHI methylase gene based on three independent observations . Both plasmids were found to be resistant to BamHI endonuclease cleavage, and chromosomal DNA isolated from E.coli HB101 cells harboring either of the plasmids pBamM6.5 or pBamM2.5 was resistant to cleavage by BamHI endonuclease . In addition, DNA isolated from lambda phage passaged through E.coli HB101 containing either plasmid was also resistant to BamHI cleavage . Expression of the BamHI methylase gene is dependent on orientation in pSP64 . In these clones preliminary evidence indicates that methylase gene expression may be under the direction of the plasmid encoded LacZ promoter.

J Exp Med, 1988 Nov 1, 168(5), 1947 - 52
The recombinant 65-kD heat shock protein of Mycobacterium bovis Bacillus Calmette-Guerin/M . tuberculosis is a target molecule for CD4+ cytotoxic T lymphocytes that lyse human monocytes; Ottenhoff TH et al.; Since little is known about Tc cells in the human immune response to intracellular parasites, we have studied the role of Tc cells in response to M . bovis Bacillus Calmette-Guerin (BCG) . Donors whose PBMC responded to BCG, purified protein derivative (PPD), and the recombinant 65-kD heat shock protein (HSP) of BCG generated BCG/PPD-specific CD4+ effector T lymphocytes that lysed PPD as well as recombinant 65-kD-pulsed monocytes . Nonpulsed or irrelevant antigen-pulsed target cells were lysed to a much lower but still significant extent . PPD-stimulated effector lymphocytes of a recombinant 65-kD nonresponder lysed PPD but not recombinant 65-kD-pulsed monocytes . Recombinant 65-kD-educated effector lymphocytes lysed both recombinant 65-kD- and PPD-pulsed monocytes . In addition, these effector cells efficiently lysed nonpulsed target cells . These results demonstrate that in recombinant 65-kD responders, the recombinant 65-kD HSP of BCG is an immunodominant target as well as a triggering molecule for BCG/PPD-specific CD4+ cytotoxic T cells that lyse autologous monocytes . The implications of these findings with respect to the role of the 65-kD HSP in autoimmunity are discussed.

Proc Natl Acad Sci U S A, 1988 Nov, 85(21), 7844 - 8
Specificity of Bacillus thuringiensis delta-endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts; Hofmann C et al.; Binding studies were performed with two 125I-labeled Bacillus thuringiensis delta-endotoxins on brush border membrane vesicles prepared from the larval midgut of the tobacco hornworm Manduca sexta or the cabbage butterfly Pieris brassicae . One delta-endotoxin, Bt2-protoxin, is a 130-kDa recombinant crystalline protein from B . thuringiensis subsp . berliner . It kills larvae of both insect species . The active Bt2-toxin is a 60-kDa proteolytic fragment of the Bt2-protoxin . It binds saturably and with high affinity to brush border membrane vesicles from the midgut of both species . The other delta-endotoxin, Bt4412-protoxin, is a 136-kDa crystalline protein from B . thuringiensis subsp . thuringiensis, which is highly toxic for P . brassicae, but not for M . sexta larvae . Bt4412-toxin, obtained after proteolytic activation of Bt4412-protoxin, shows high-affinity saturable binding to P . brassicae vesicles but not to M . sexta vesicles . The correlation between toxicity and specific binding is further strengthened by competition studies . Other B . thuringiensis delta-endotoxins active against M . sexta compete for binding of 125I-labeled Bt2-toxin to M . sexta vesicles, whereas toxins active against dipteran or coleopteran larvae do not compete . Bt2-toxin and Bt4412-toxin bind to different sites on P . brassicae vesicles.

Curr Genet, 1988 Nov, 14(5), 493 - 500
Initiation of rrn transcription in chloroplasts of Euglena gracilis bacillaris; McGarvey P et al.; The site of initiation of chloroplast rRNA synthesis was determined by S1-mapping and by sequencing primary rRNA transcripts specifically labeled at their 5'-end . Transcription initiates at a single site 53 nucleotides upstream of the 5'-end of the mature 16S rRNA under all growth conditions examined . The initiation site is within a DNA sequence that is highly homologous to and probably derived from a tRNA gene-region located elsewhere in the chloroplast genome . A nearly identical sequence (102 of 103 nucleotides) is present near the replication origin . The near identity of the two sequences suggests a common mode for control of transcription of the rRNA genes and initiation of chloroplast DNA replication . The related sequence in the tRNA gene-region does not appear to serve as a transcript initiation site.

Mikrobiologiia, 1988 Nov-Dec, 57(6), 1017 - 23
{The cell wall of Bacillus brevis producing gramicidin S, a cyclodecapeptide antibiotic}; Ostrovskii DN et al.; Gramicidin S is tritiated in Bacillus brevis G.-B . intact cells by activated tritium atoms (which is indicative of its surface localisation) . The cell wall protein is tritiated very weakly, which shows that it is screened, apparently, by gramicidin adsorbed on its surface . The cell wall protein is not a glycoprotein and its interaction with exogenous gramicidin S causes cell aggregation . As follows from the Rf value after the chromatographic separation of B . brevis lipids, the reaction of staining, and the data of H-NMR spectroscopy for the fraction of phospholipids, the main membrane phospholipid is an anionic acetylated phosphatidyl ethanolamine.

Am Rev Respir Dis, 1988 Nov, 138(5), 1124 - 8
Use of allophycocyanin allows quantitative description by flow cytometry of alveolar macrophage surface antigens present in low numbers of cells; Fuchs HJ et al.; Autofluorescence has limited quantitative description by flow cytometry of certain alveolar macrophage surface antigens . Autofluorescence is most significant when macrophages are excited by blue light and emission is monitored in the green-yellow light range . Because of recent advances in flow cytometry and in the development of fluorescent labeling compounds, such as allophycocyanin, autofluorescence can be minimized by the use of red excitation and emission wavelengths . We have developed a three-step labeling technique that discretely identifies the minor subpopulation of normal rat alveolar macrophages that express Class II major histocompatibility (Ia) antigens . Using this technique, we observed that alveolar macrophage Ia antigen expression increases at least fourfold after intravenous administration of bacillus Calmette-Guerin to previously primed rats . Additionally, we observed that in vitro treatment of alveolar macrophages by exposure to gamma interferon results in an increase in Ia antigen expression . This new method is significant because flow cytometry is a powerful tool with which to analyze quickly, accurately, and reproducibly alveolar macrophage surface antigens.

Am J Trop Med Hyg, 1988 Nov, 39(5), 427 - 33
Common surface epitope of Bartonella bacilliformis and Chlamydia psittaci; Knobloch J et al.; A serosurvey revealed intense cross-reactivity between Bartonella bacilliformis and Chlamydia psittaci . One of the cross-reacting Bartonella antigens was identified as lipopolysaccharide which reacted with Bartonella as well as with Chlamydia serum antibodies . A monoclonal Bartonella antibody bound to Bartonella lipopolysaccharide as well as to the surfaces of Bartonella bacilliformis and Chlamydia psittaci . It was thus demonstrated that Chlamydia psittaci carries a surface epitope identical to an epitope of Bartonella lipopolysaccharide . The lipopolysaccharide was preliminarily characterized by polyacrylamide gel electrophoresis and by a lectin-binding assay . The lipopolysaccharides of Bartonella bacilliformis and Chlamydia psittaci are not identical.

J Mol Biol, 1988 Oct 20, 203(4), 1097 - 118
Coenzyme-induced conformational changes in glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus; Skarzynski T et al.; The structure of apo-glyceraldehyde-3-phosphate dehydrogenase (GAPDHase) from Bacillus stearothermophilus has been refined using a restrained least-squares method . The final crystallographic R-factor is 0.177 for all 53,315 reflections between 7.0 and 2.5 A . The resulting model has been analysed with respect to lattice interactions, molecular symmetry, temperature factors and solvent structure showing that, apart from local deviations due to intermolecular contact, the molecule exhibits a very high degree of local 222 symmetry . Analysis of differences between the structure of apo-GAPDHase and the previously refined holo-GAPDHase at 1.8 A resolution reveals details of conformational change in the enzyme induced by cofactor binding . The change, which was previously described as a rigid-body rotation of the coenzyme-binding domain with respect to the catalytic domain, is of more complex nature and involves relative shifts of several structural elements in the coenzyme-binding domain and some small changes in the catalytic domain . A possible mechanism of this conformational change is proposed based on the comparison of the refined structures and model-building studies . According to this mechanism, the adenosine moiety of NAD can initially bind to the protein in the apo-enzyme conformation . Several attractive interactions resulting from the initial binding of the coenzyme trigger conformational changes in the molecule of GAPDHase that: (1) create the productive nicotinamide-moiety binding site; (2) improve enzyme-coenzyme interactions at the adenosine moiety; (3) modify the active site to optimize the positioning of catalytic residues and ion-binding sites . Implications of the proposed mechanism for existing experimental data on binding of NAD analogues to GAPDHase are discussed.

Nature, 1988 Oct 20, 335(6192), 740 - 3
Stabilization of protein structure by interaction of alpha-helix dipole with a charged side chain; Sali D et al.; The alpha-helix in proteins has a dipole moment resulting from the alignment of dipoles of the peptide bond which can perturb the pKas of ionizing groups . One of the two histidine residues (His18) in barnase, the small ribonuclease from Bacillus amyloliquefaciens, is located at the negatively charged end (C-terminal) of an alpha-helix . From NMR titrations of wild-type and engineered mutants we find that the pKa of His18 is 7.9 in wild-type enzyme, 1.6 units above the value in the urea-denatured enzyme and in model peptides . This implies that there is a favourable interaction between the protonated form of His18 and the alpha-helix that should stabilize the native structure at neutral pH by 2.1 kcal mol-1 . Denaturation at various values of pH of wild-type and muant enzymes engineered at position 18 shows that this is so . The increase in stability of the enzyme as the pH changes from 8.5 to 6.3 is attributable to this interaction, and the pH-stability curve fits pKa values for His18 in native and urea-denatured enzymes that are consistent with the NMR data.

J Immunol, 1988 Oct 15, 141(8), 2729 - 33
Gene isolation with human T lymphocyte probes . Isolation of a gene that expresses an epitope recognized by T cells specific for Mycobacterium bovis BCG and pathogenic mycobacteria; Mustafa AS et al.; We have used human CD4+ T lymphocyte clones as primary probes to identify and isolate lambda gt11 rDNA clones that express epitopes recognized by T cells . The method that we describe here permits a direct survey of T cell epitope coding sequences in genomic DNA or cDNA libraries . A lambda gt11 library of Mycobacterium leprae DNA was screened with M . leprae-reactive human T cell clones as probes, allowing the isolation of a M . leprae DNA clone encoding the unidentified Ag . This DNA clone differs in restriction maps from those previously identified by antibody probes and encodes an epitope that is unique to vaccine strains of Mycobacterium bovis bacillus Calmette-Guerin and pathogenic mycobacteria . This method is generally applicable and should expedite the study of Ag and epitopes important to the T cell response in infections and in autoimmune diseases.

Biochem Biophys Res Commun, 1988 Oct 14, 156(1), 537 - 42
Site-directed mutagenesis at the regulatory site of fructose 6-phosphate-1-kinase from Bacillus stearothermophilus; Valdez BC et al.; We have mutated Arg-25, Asp-59 and Arg-211 to alanine; and Asp-59 also to methionine, in fructose 6-phosphate-1-kinase from B . stearothermophilus (designated as RA25, DA59, RA211 and DM59 respectively) . All four mutants did not change the affinity of the enzyme for ATP . RA25 has half the affinity for fructose 6-phosphate and exhibits sigmoidicity with respect to this substrate (Hill # = 2.0) . DA59 has the same affinity for phosphoenolpyruvate (PEP) as the wild type whereas DM59 has 3-fold the affinity for this modulator and the inhibition is reversed by GDP . RA25 and RA211 are 100-fold less sensitive to PEP inhibition which is not relieved by GDP . It is concluded that Arg-25 and Arg-211, but not Asp-59, are involved in the direct binding of PEP and GDP.

J Biol Chem, 1988 Oct 5, 263(28), 14368 - 73
The solubilization of tetrameric alkaline phosphatase from human liver and its conversion into various forms by phosphatidylinositol phospholipase C or proteolysis; Hawrylak K et al.; When membrane-bound human liver alkaline phosphatase was treated with a phosphatidylinositol (PI) phospholipase C obtained from Bacillus cereus, or with the proteases ficin and bromelain, the enzyme released was dimeric . Butanol extraction of the plasma membranes at pH 7.6 yielded a water-soluble, aggregated form that PI phospholipase C could also convert to dimers . When the membrane-bound enzyme was solubilized with a non-ionic detergent (Nonidet P-40), it had the Mr of a tetramer; this, too, was convertible to dimers with PI phospholipase C or a protease . Butanol extraction of whole liver tissue at pH 6.6 and subsequent purification yielded a dimeric enzyme on electrophoresis under nondenaturing conditions, whereas butanol extraction at pH values of 7.6 or above and subsequent purification by immunoaffinity chromatography yielded an enzyme with a native Mr twice that of the dimeric form . This high molecular weight form showed a single Coomassie-stained band (Mr = 83,000) on electrophoresis under denaturing conditions in sodium dodecyl sulfate, as did its PI phospholipase C cleaved product; this Mr was the same as that obtained with the enzyme purified from whole liver using butanol extraction at pH 6.6 . These results are highly suggestive of the presence of a butanol-activated endogenous enzyme activity (possibly a phospholipase) that is optimally active at an acidic pH . Inhibition of this activity by maintaining an alkaline pH during extraction and purification results in a tetrameric enzyme . Alkaline phosphatase, whether released by phosphatidylinositol (PI) phospholipase C or protease treatment of intact plasma membranes, or purified in a dimeric form, would not adsorb to a hydrophobic medium . PI phospholipase C treatment of alkaline phosphatase solubilized from plasma membranes by either detergent or butanol at pH 7.6 yielded a dimeric enzyme that did not absorb to the hydrophobic medium, whereas the untreated preparations did . This adsorbed activity was readily released by detergent . Likewise, alkaline phosphatase solubilized from plasma membranes by butanol extraction at pH 7.6 would incorporate into phosphatidylcholine liposomes, whereas the enzyme released from the membranes by PI phospholipase C would not incorporate . The dimeric enzyme purified from a butanol extract of whole liver tissue carried out at pH 6.6 did not incorporate . We conclude that PI phospholipase C converts a hydrophobic tetramer of alkaline phosphatase into hydrophilic dimers through removal of the 1,2-diacylglycerol moiety of phosphatidylinositol . Based on these and others' findings, we devised a model of alkaline phosphatase's conversion into its various forms.

Int J Food Microbiol, 1988 Oct, 7(2), 109 - 14
The evaluation of the recovery capacity of media for heat-treated Bacillus stearothermophilus spore strips; Brown GD et al.; Six media were assessed for their capacity to recover spores of Bacillus stearothermophilus from three commercially available types of spore strip heated for up to 5 min at 121 degrees C in steam . All media recovered similar numbers of spores from unheated strips; however, there was considerable difference in survivor counts when the media were used to recover heated spores . Media containing starch consistently recovered the highest number of spores . The greatest difference in recovery capacity between the media was observed when spore strips were heated for 5 min at 121 degrees C.

Indian J Lepr, 1988 Oct, 60(4), 581 - 8
Pattern of relapses in pauci-bacillary leprosy patients treated with M.D.T . (W.H.O . 1982); Reddy PK et al.; Out of 92 Pauci-bacillary leprosy patients treated with MDT (WHO 1982), two patients developed indisputable clinical signs of relapse during 10th and 26th month of observation period . Two more patients developed reversal reaction during 8th and 12th month of observation period, which we presume to be early manifestation of relapse . Addition of a more bactericidal drug, rifampicin, appear to have a bearing on the incidence of relapse, though not on it's incubation period . No change of classification was noticed at the time of relapse . Incidence of relapse in female patients was much higher than in male patients.

Rev Argent Microbiol, 1988 Oct-Dec, 20(4), 163 - 70
Evaluation of circulating immune complexes in leprosy; Ramos GS et al.; Circulating immune complexes (CIC) were evaluated in leprosy by 4 methods: the 125I-C1q binding assay (C1q), the platelet aggregation test (PAT), the 3.5% polyethylene glycol (PEG) precipitation test and the 2.5% PEG precipitation assay . Serum samples belonged to lepromatous leprosy bacilloscopy positive (LL+), lepromatous leprosy bacilloscopy negative (LL-), tuberculoid (TT) and first grade contact group (Co) . Studies performed by the 3 first methods showed higher CIC levels in LL+ group (p less than 0.01) and lower values in the 3 others, all of them when compared to normals . On the contrary, the 2.5% PEG precipitation test gave less discriminative results giving only p less than 0.01 in LL+ . CIC values obtained in the contact group showed significant results compared to normals but similar to LL- and TT groups . The C1q binding assay and the PAT were the most discriminative methods giving r = 0.90; C1q versus 3.5% PEG, r = 0.36; C1q vs 2.5% PEG, r = 0.14 . The PAT compared to 3.5% PEG, r = 0.48 and PAT vs . 2.5% PEG, r = 0.24 . Therefore it may be concluded as follows: a) The radioiodinated C1q binding assay and the PAT are recommended for the study of CIC in leprosy; b) The 2.5% PEG precipitation assay offers less sensitivity since it gave similar value in LL-, TT, Co and controls; c) CIC levels observed in LL+ patients may be induced by the antigenic overload demonstrated by the positive bacilloscopy; d) The contacts have CIC levels significantly different from the normal population possibly caused by a subclinical infection.

Jikken Dobutsu, 1988 Oct, 37(4), 447 - 53
Pathogenicities of two CAR bacillus strains in mice and rats; Shoji Y et al.; Two strains of CAR bacillus from a mouse (CB-M) and a rat (CB-R) which were passaged 11th in embryonated chicken eggs via the allantoic route were inoculated intranasally in ICR mice and Wistar rats . The histopathological changes and the localization of the CAR bacillus in the tracheas and lungs of these animals were investigated microscopically 2, 4 and 8 weeks postinoculation (PI) . Histopathological changes similar to those in natural cases of CAR bacillus infection, showing severe peribronchial lymphoid cuffing, were first recognized 4 weeks PI . CAR bacillus was also found on the cilia of the respiratory epithelium . These histopathological changes were more remarkable in mice inoculated with CB-M and in rats inoculated with CB-R.

Mol Gen Genet, 1988 Oct, 214(2), 343 - 7
Correlation of physical maps and some genetic functions in the genomes of the kappa-theta phage family of Bacillus licheniformis; Doskocil J et al.; Natural recombinant genomes between several, phenotypically distinct forms of phages kappa and theta were isolated and characterized by DNA restriction fragment mapping and electron microscopic heteroduplex analysis . The phenotypes of the recombinants were correlated with the physical maps of the genomes, and several genetic functions were therefore defined and mapped . All genes necessary for the assembly of infectious virus particles map in a contiguous tract of DNA comprising about 20 kb, or nearly one third of the genome length . No DNA homology occurs within these domains of the two genomes, so that homologous recombination does not take place here and phenotypic mixing of the phages is eo ipso excluded . Other regions of heterology contain regulatory genes responsible for the lytic or temperature character of the phages, and for exclusion of phage kappa by 9.

Appl Environ Microbiol, 1988 Oct, 54(10), 2515 - 20
Heat shock affects permeability and resistance of Bacillus stearothermophilus spores; Beaman TC et al.; Heat shock of dormant spores of Bacillus stearothermophilus ATCC 7953 at 100 or 80 degrees C for short times, the so-called activation or breaking of dormancy, was investigated by separating the resulting spores by buoyant density centrifugation into a band at 1.240 g/ml that was distinct from another band at 1.340 g/ml, the same density as the original spores . The proportion of spores at 1.240 g/ml became larger when the original dormant spores were heated for a longer period of time, but integument-stripped dormant spores were quickly and completely converted to spores with a band at 1.240 g/ml . The spores with bands at both 1.240 and 1.340 g/ml were germinable faster than the original dormant spores and thus were considered to be activated . The spores with a band at 1.240 g/ml, which were considered to be fully activated, were apparently permeabilized, with a resulting complete depletion of dipicolinic acid, partial depletion of minerals, susceptibility to lysozyme action, permeation of the gradient medium, changed structural appearance in electron micrographs of thin-sectioned spores, and partly decreased heat resistance (D100 = 453 min) compared with the original dormant spores (D100 = 760 min) . However, the fully activated spores with a band at 1.240 g/ml, although devoid of dipicolinic acid, still were much more resistant than germinated spores or vegetative cells (D100 = 0.1 min) . The spores with a band at 1.340 g/ml, which were considered to be partly activated, showed no evidence of permeabilization and were much more heat resistant (D100 = 1,960 min) than the original dormant spores.(ABSTRACT TRUNCATED AT 250 WORDS)

Kidney Int, 1988 Oct, 34(4), 467 - 73
Lupus nephropathy in New Zealand F1 hybrid mice treated by (-)15-deoxyspergualin; Okubo M et al.; The effect of (-)15-deoxyspergualin (15-dsp), a new immunosuppressant which was originally separated from the culture filtrate of a strain of Bacillus laterosporus, was evaluated in this study . Various doses of 15-dsp were subcutaneously administered to New Zealand black/white F1 hybrid mice (B/WF1) four times a week starting at 14 weeks of age, just prior to the onset of nephropathy . The life span of the treated animals, studied at 0.6 to 6.0 mg/kg body weights, compared with the control mice was significantly prolonged by 15-dsp treatment (percent survival of the treated mice at 50 to 70 weeks of age was significantly higher than that of the control mice, except that of the 0.6 mg group at 60 wks of age, P less than 0.05 by Fisher's exact test) . In the 6.0 mg group of mice, complete suppression of spontaneously progressive splenomegaly with decreased total spleen cells was observed at 24 through 36 weeks of age compared with the same-aged control group of mice (P less than 0.01) . Absolute numbers of L3T4+ splenocytes determined by flow cytometry, as well as L3T4+/Lyt2+ ratio, were decreased, while in vitro interleukin 2 production by splenocytes induced with staphylococcal enterotoxin A was significantly enhanced . Serum IgG anti-ds DNA antibody levels, measured by radioimmunoassay in the treated mice, were significantly lower at 24 through 36 weeks of age than those in the control mice (P less than 0.01), and the incidence of significant proteinuria (greater than or equal to 100 mg/dl) in the 15-dsp group was lower at both 32 and 36 weeks of age (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Gynecol Oncol, 1988 Oct, 31(2), 321 - 6
Disseminated granulomatous disease (BCGosis) following chemoimmunotherapy for ovarian carcinoma; Kelleher MB et al.; The use of BCG (bacille Calmette Guerin) in conjunction with cytotoxic chemotherapeutic agents has been advocated in patients with ovarian carcinoma . We describe a patient with stage III, grade I, endometrioid carcinoma of the ovary treated with cisplatin, doxorubicin, cyclophosphamide, and BCG . Following one course of therapy she presented with an elevated temperature, purpuric skin rash, abnormal liver function tests and hematological indices, and multiple organ failure resulting in sepsis and death . At autopsy, disseminated noncaseating granulomas were found in the lungs, hilar lymph nodes, liver, and spleen . Metastatic carcinoma was not present in these organs . This report describes the rapid onset of a disseminated BCG infection (BCGosis) in a patient with ovarian carcinoma receiving chemoimmunotherapy . Clinical recognition of BCGosis in immunocompromised patients is difficult but should be considered in the differential diagnosis of patients presenting with unexplained febrile illness, functional abnormalities in multiple organ systems, and a history of immunotherapy with BCG . Appropriate specimen collection is emphasized.

Eur J Biochem, 1988 Oct 1, 176(3), 559 - 65
The function of beta-N-acetyl-D-glucosaminyl monophosphorylundecaprenol in biosynthesis of lipoteichoic acids in a group of Bacillus strains; Shimada A et al.; Membrane preparations, obtained from Bacillus strains which have N-acetylglucosamine-linked lipoteichoic acids in their membranes, were shown to catalyze the transfer of N-{14C}acetylglucosamine (GlcNAc) from beta-{14C}GlcNAc-P-undecaprenol to endogenous polymer . In this reaction, alpha-GlcNAc-P-undecaprenol or alpha-GlcNAc-PP-undecaprenol could not substitute for beta-GlcNAc-P-undecaprenol as the N-acetylglucosamine donor . This enzyme was most active at pH 6.0 and in the presence of 40 mM MgCl2 . The apparent Km for beta-GlcNAc-P-undecaprenol was 2 microM . The radioactive polymer products, solubilized by hot phenol treatment, coincided with lipoteichoic acids in chromatographic behavior . Hydrogen fluoride treatment of the polymer products gave a major fragment identical with GlcNAc(alpha 1----2)glycerol, which corresponded to the dephosphorylated repeating units of the lipoteichoic acids in the examined strains . Thus it is concluded that beta-GlcNAc-P-undecaprenol serves as the donor of N-acetylglucosamine in the biosynthesis of lipoteichoic acids in a group of Bacillus strains.

Vet Microbiol, 1988 Oct, 18(2), 191 - 6
Identification of Mycobacterium bovis isolates using a monoclonal antibody; Corner LA et al.; A rapid immunoperoxidase slide assay for the identification of Mycobacterium bovis culture isolates is described . The monoclonal antibody used in this assay is specific for the M . tuberculosis complex of organisms . All M . bovis isolates tested, including 151 separate field isolates of M . bovis were positive as were 11 out of 12 M . tuberculosis strains and 4 out of 6 Bacillus Calmette Guerin (BCG) strains . One strain each of M . africanum and M . microti was negative . This assay provides a considerable improvement in both time and expense over the conventional methods of biochemical typing of M . bovis.

J Bacteriol, 1988 Oct, 170(10), 4942 - 5
Integration and mapping of Bacillus megaterium genes which code for small, acid-soluble spore proteins and their protease; Sussman MD et al.; Four genes (ssp genes) coding for small, acid-soluble spore proteins of Bacillus megaterium and the gene for the protease that cleaves them during germination were cloned in the integratable plasmid pJH101 . Each plasmid was integrated into the B . megaterium chromosome by a Campbell-type mechanism, allowing mapping of all five genes . The gene for the small, acid-soluble spore protein-specific protease (gpr) mapped near rib, and the sspA gene mapped between argA and hisA . The three other genes of the spp gene family (sspB, -D, and -F) all mapped near metC/D, with the order: sspF-sspD-metC/D-hemA-argO-sspB . While neither gpr nor sspF has been mapped in B . subtilis, the positions of the sspA, -B, and -D loci are similar in B . megaterium and B . subtilis, suggesting that the members of this multigene family have not recently undergone significant movement on the chromosome . It appears that more gene rearrangement has occurred in the flanking genes than has occurred in the ssp family of genes producing the small, acid-soluble spore proteins.

Indian J Lepr, 1988 Oct, 60(4), 499 - 505
A double-blind controlled clinical trial to assess the role of anti-histamines in the treatment of multi-bacillary leprosy; Thomas A et al.; A double blind controlled clinical trial to assess the role of anti-histamines as a supplement in the treatment of leprosy was conducted in multi-bacillary cases of leprosy . In all, 120 patients with lepromatous or borderline leprosy were randomly allocated to a regimen of clofazimine and dapsone for 12 months with or without a supplement of pheniramine maleate for the first 3 months . During the 12-month period, 92% of the patients who received the supplement and 86% of the patients who had not received it had moderate or marked clinical improvement . The BI values decreased from 4.1 to 3.4 and 4.2 to 3.3, respectively . The results over the 12-month period showed that the addition of the antihistamine had not enhanced the efficacy of the regimen as evidenced by clinical and bacteriological findings.

Mol Gen Genet, 1988 Oct, 214(2), 241 - 8
Cloning, sequencing and expression of a Bacillus bacteriolytic enzyme in Escherichia coli; Potvin C et al.; Several hundred bacterial isolates were screened for bacteriolytic activity by growing them on agar medium containing autoclaved, lyophilized Micrococcus lysodeikticus cells as the substrate . A Bacillus sp . producing the largest lytic zone was selected . A genomic bank of this selected bacterium was constructed in the multi-functional vector pTZ18R, with partial SauIIIA DNA fragments inserted at the SalI restriction site . Screening of 800 colonies of this bank for cell lysis gave 5 recombinants exhibiting lytic activity, as detected by analysis of extracts of sonicated Escherichia coli cells on denaturing polyacrylamide gels containing autoclaved, lyophilized M . lysodeikticus cells as the substrate . One clone (pBH2500), expressed in E . coli strain NM522, was found to code for a lytic enzyme corresponding, in molecular weight, to the 27 kDa Bacillus sp . hydrolase . This clone with an insertion of 2.5 kb was then subcloned as a 929 bp EcoRI-SauIIIA fragment in pTZ18R (pBH929) and showed higher cell lytic activity . A unique open reading frame for a protein of 251 amino acids, followed by a putative terminator sequence, was found after a consensus ribosome binding site . A putative leader sequence was identified in the first 37 amino acids . One truncated subclone (pBH703), corresponding to 196 out of 251 residues from the protein N-terminal end, still possessed lytic activity.

Biochem J, 1988 Oct 1, 255(1), 365 - 8
Probable detection of kil peptide derived from colicin E1 plasmid in the envelope fraction of Escherichia coli HB101 carrying pEAP31; Aono R; Escherichia coli carrying plasmid pEAP31 produces extracellularly alkalophilic Bacillus penicillinase encoded on the plasmid . The extracellular production has been suggested to be caused by activation of dormant colicin E1 kil gene . Two peptides that could be respectively precursor and mature products of colicin E1 kil gene were detected on an SDS/polyacrylamide gel . One of the peptides (Mr 4800), which was probably a precursor peptide, was detected in the inner-membrane fraction from the organism when envelope proteins were subjected to differential solubilization . The other (Mr 3500), which was a mature peptide, was detected in the outer-membrane fraction of the organism . The mature peptide was only detected in the envelope of cells releasing the penicillinase transiently accumulated in the periplasm into the culture medium.

J Bacteriol, 1988 Oct, 170(10), 4732 - 8
Molecular characterization of a gene encoding a 72-kilodalton mosquito-toxic crystal protein from Bacillus thuringiensis subsp . israelensis; Donovan WP et al.; A gene encoding a 72,357-dalton (Da) crystal protein of Bacillus thuringiensis var . israelensis was isolated from a native 75-MDa plasmid by the use of a gene-specific oligonucleotide probe . Bacillus megaterium cells harboring the cloned gene (cryD) produced significant amounts of the 72-kDa protein (CryD), and the cells were highly toxic to mosquito larvae . In contrast, cryD-containing Escherichia coli cells did not produce detectable levels of the 72-kDa CryD protein . The sequence of the CryD protein, as deduced from the sequence of the cryD gene, was found to contain regions of homology with two previously described B . thuringiensis crystal proteins: a 73-kDa coleopteran-toxic protein and a 66-kDa lepidopteran- and dipteran-toxic protein of B . thuringiensis subsp . kurstaki . A second gene encoding the B . thuringiensis subsp . israelensis 28-kDa crystal protein was located approximately 1.5 kilobases upstream from and in the opposite orientation to the cryD gene.

J Bacteriol, 1988 Oct, 170(10), 4669 - 74
Organization of the biosynthesis genes for the peptide antibiotic gramicidin S; Krause M et al.; A recombinant bacteriophage containing the intact Bacillus brevis gene for gramicidin S synthetase 1, grsA, and the 5' end of the gramicidin S synthetase 2 gene, grsB, was identified by screening an EMBL3 library with anti-GrsA antibodies . This clone, EMBL315, has a 14-kilobase (kb) insert that hybridizes to the previously isolated 3.9-kb fragment of the grsB gene, which encodes the 155-kilodalton ornithine-activating domain of gramicidin S synthetase 2 . Deletion and subcloning experiments with the 14-kb insert located the grsA structural gene and its putative promoter on a 4.5-kb PvuII fragment which encoded the full-length 120-kilodalton protein in Escherichia coli . In addition, hybridization analysis revealed that the 5' end of the grsB gene is located approximately 3 kb from the grsA structural gene . Furthermore, these studies indicated that grsA and grsB are transcribed in opposite orientations.

Biochemistry, 1988 Sep 20, 27(19), 7390 - 4
Assignment of histidine resonances in the 1H NMR (500 MHz) spectrum of subtilisin BPN' using site-directed mutagenesis; Bycroft M et al.; A spin-echo pulse sequence has been used to resolve the six histidine C-2H protons in the 500-MHz NMR spectrum of subtilisin BPN' . Five of these residues have been substituted by site-directed mutagenesis, and this has enabled a complete assignment of these protons to be obtained . Analysis of the pH titration curves of these signals has provided microscopic pKas for the six histidines in this enzyme . The pKas of the histidine residues in subtilisin BPN' have been compared with the values obtained for the histidines in the homologous enzyme from Bacillus licheniformis (subtilisin Carlsberg) . Four of the five conserved histidines titrate with essentially identical pKa's in the two enzymes . It therefore appears that the assignments made for these residues in subtilisin BPN' can be transferred to subtilisin Carlsberg . On the basis of these assignments, the one histidine that titrates with a substantially different pKa in the two enzymes can be assigned to histidine-238 . This difference in pKa has been attributed to a Trp to Lys substitution at position 241 in subtilisin Carlsberg.

Ann Intern Med, 1988 Sep 15, 109(6), 449 - 55
Cutaneous vascular lesions and disseminated cat-scratch disease in patients with the acquired immunodeficiency syndrome (AIDS) and AIDS-related complex; Koehler JE et al.; Cutaneous lesions develop frequently in patients infected with human immunodeficiency virus (HIV) . We describe the clinical features of four patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex who developed angiomatous nodules involving skin and bone, 2 of whom were scratched by a cat . Some of these lesions were clinically indistinguishable from Kaposi sarcoma . When examined with Warthin-Starry staining and electron microscopy, these nodules were noted to contain numerous clumps of a bacterium . Immunoperoxidase staining with an antiserum raised against the cat-scratch disease bacillus stained these organisms in all patients . Cat-scratch disease is usually a self-limited infection, but complicated or prolonged infections have been described in both normal and immunocompromised hosts . In our patients infected with HIV, manifestations of systemic cat-scratch disease included angiomatous nodules, severe systemic symptoms of fever, chills, night sweats and weight loss, elevated erythrocyte sedimentation rate, and decreased hematocrit . Cutaneous lesions involved the face, trunk, and extremities and numbered 2 to greater than 60; osseous lesions involved the fibula, radius, femur, and tibia, and were present in two of four patients . Treatment with x-ray therapy, intralesional vinblastine, penicillin, dicloxacillin, cephradine, and nafcillin had no effect on any lesions; however, treatment with erythromycin, doxycycline, or antimycobacterial antibiotics resulted in complete and rapid resolution of the cutaneous and osseous lesions, and the accompanying signs and symptoms of systemic infection . In patients with AIDS or AIDS-related complex, angiomatous nodules should be carefully evaluated for the presence of this organism, which can be treated and cured with antibiotic agents.

J Immunol, 1988 Sep 15, 141(6), 1813 - 8
Recognition by cytotoxic T lymphocytes of Qa-2 antigens . Sensitivity of Qa-2 molecules to phosphatidylinositol-specific phospholipase C; Mann D et al.; Con A splenic lymphoblasts were incubated with phosphatidyl-inositol specific phospholipase C (PIPLC) derived from Bacillus thuringiensis and subsequently analyzed for Qa-2 Ag with the Qa-2 reactive mAb Qa-m2 . This treatment completely removed Qa-2 detectable Ag on lymphoblasts from H-2d animals, indicating that these molecules are likely anchored to the cell membrane through phosphatidyl inositol (PI) . Although exposure of lymphoblasts from H-2b mice to PIPLC greatly reduced Qa-2 expression, a subpopulation of cells retained a limited quantity of the Ag . Bulk cultured anti-Qa-2 CTL generated against the Qa-2 region from H-2b haplotype mice lysed Qa-2+ targets from B6.K2 (H-2b) and BALB/cJ (H-2d) animals . Pretreatment of these lymphoblast targets with PIPLC completely abolished lysis of the BALB/cJ target cells, whereas lysis of B6 targets was reduced only slightly . Anti-Qa-2 CTL clones tested against PIPLC-treated B6 target cells revealed two patterns of reactivity . One group of clones was unaffected in its ability to lyse PIPLC-pretreated targets and cross-reacted on Q6d/Ld molecules expressed on transfected L cells . A second group was unable to lyse PIPLC-pretreated lymphoblasts and cross-reacted on Q7d/Ld targets . These data suggest that H-2b-derived lymphoblasts express two different types of Qa-2 molecules with respect to PIPLC sensitivity; one type is sensitive to PIPLC and cross-reactive with Q7d, the other type is resistant to PIPLC and cross-reactive with Q6d . In contrast, H-2d lymphoblasts express only the PIPLC-sensitive type of molecules . It was also noted that bulk cultured anti-Qa-2 CTL more readily lysed H-2b target cells expressing a smaller quantity of PIPLC-resistant Ag than H-2d targets expressing a larger amount of PIPLC-sensitive Ag . Further, anti-Qa-2 CTL clones readily lysed PIPLC-treated target cells expressing very low levels of serologically detectable Qa-2 . This suggests that recognition of class I molecules anchored to the membrane via a PIPLC-resistant linkage may more readily activate CTL for expression of lytic activity than molecules anchored through PI.

J Clin Oncol, 1988 Sep, 6(9), 1450 - 5
Bacillus Calmette-Guérin therapy alters the progression of superficial bladder cancer; Herr HW et al.; The effectiveness of BCG in preventing disease progression in patients with superficial bladder cancer is evaluated . Long-term follow-up of high-risk patients treated in a previously reported randomized control trial of intravesical plus percutaneous BCG shows that progression occurred in 41/43 (95%) of control and 23/43 (53%) of BCG-treated patients . Muscle invasive and/or metastatic disease occurred with equal frequency in the two groups, but was significantly delayed by BCG treatment (P = .012) . Cystectomies were required in 18/43 (42%) control and 11/43 (26%) BCG-treated patients . Median time to cystectomy was 8 months for control v 24 months for BCG-treated patients . Based on initial treatment, survival was improved by BCG therapy (P = .032) (median follow-up 6 years) . These results suggest that in high-risk patients intravesical BCG can delay disease progression, prolong the period of bladder preservation, and increase overall survival.

J Urol, 1988 Sep, 140(3), 647 - 50
Tumor-specific immunotherapy of murine bladder cancer with butanol-extracted antigens and ethylchlorformate polymerized tumor protein; Rochester MG et al.; Successful treatment of superficial bladder cancer using nonspecific immunotherapy with Bacillus Calmette-Guerin (BCG) has been well documented . Investigation of two potential tumor-specific immunotherapeutic agents using a murine transitional-cell carcinoma model (MBT-2) is reported . The survival of mice immunized with tumor proteins obtained by treating tumor cells with either 1-butanol or ethylchlorformate was compared to the survival of animals immunized with BCG . Long-term immunity conferred by each of these agents was also assessed . Significant protection by both agents was noted in all treatment groups compared to controls . Long-term immunity was also found to result from treatment with both investigational agents as well as with BCG . Butanol-extracted antigens and ethylchlorformate polymerized tumor protein may be useful as immunotherapeutic alternatives to BCG.

J Urol, 1988 Sep, 140(3), 498 - 500
Prophylactic maltose tetrapalmitate and bacillus Calmette-Guerin immunotherapy of recurrent superficial bladder tumors: preliminary report; Ibrahiem EH et al.; We divided randomly into 3 groups 47 patients with recurrent superficial transitional cell carcinoma of the bladder: group 1-15 controls who underwent transurethral resection only, group 2-17 patients who underwent transurethral resection and bacillus Calmette-Guerin therapy, and group 3-15 patients treated by transurethral resection and maltose tetrapalmitate . Mean followup was 22.93 months for the controls, 28.0 months for group 2 and 24.4 months for group 3 . The recurrence rate per 100 patient-months was 11.34 in the controls, 7.4 in group 2 and 7.19 in group 3, and the recurrence index per month was 0.113, 0.070 and 0.072, respectively . The recurrence rate and recurrence index per month were significantly decreased in the treated groups compared to the controls (p less than 0.005) . There was no significant difference between the bacillus Calmette-Guerin and maltose tetrapalmitate groups . Invasive carcinoma developed in 60 per cent of the controls, 29.4 per cent of group 2 and 20 per cent of group 3 . Invasive carcinoma required cystectomy or definitive radiotherapy . Bacillus Calmette-Guerin caused irritation of the bladder mucosa, while maltose tetrapalmitate did not have any side effects.

J Bacteriol, 1988 Sep, 170(9), 4113 - 8
Nutrient-stimulated methylation of a membrane protein in Bacillus licheniformis; Bernlohr RW et al.; When nitrogen-starved vegetative cells of Bacillus licheniformis A5 were presented with a good nitrogen source in the presence of chloramphenicol and methyl-labeled methionine, a 40-kilodalton (kDa) protein was found to be reversibly methylated, with a half-life of approximately 10 to 15 min . The 40-kDa protein was strongly methylated in response to the addition of ammonia, glutamine, or sodium glutamate nitrogen sources that produce generation times of less than or equal to 90 min) but was very poorly methylated in the absence of a nitrogen source or in the presence of potassium glutamate or histidine (generation times of greater than 150 min) . The methylated protein was found to be membrane associated, but the methylation reaction did not appear to be related to chemotaxis, because the spectrum of nutrients that promoted methylation was different from that which prompted a chemotactic response . In addition, the methyl residue on the 40-kDa protein was found to be alkali stable . Approximately 180 to 640 molecules of the methylated protein were found per cell . The characteristics of this methylated protein were consistent with the hypothesis that the reversible methylation of the protein functions in nutrient sensing to regulate growth, cell division, and the initiation of sporulation.

Mikrobiologiia, 1988 Sep-Oct, 57(5), 734 - 9
{Regulation of alpha-amylase biosynthesis in Bacillus diastaticus mutants with various levels of enzyme synthesis}; Murygina VP; The regulation of alpha-amylase biosynthesis was studied in Bacillus diastaticus mutants with different levels of the enzyme synthesis (by two orders of magnitude) . The enzyme biosynthesis was shown to be regulated by induction and catabolite repression . Maltose, starch and methyl-alpha,D-glucoside (which cannot be metabolised) induced the synthesis while glucose and fructose acted as catabolite repressors.

Mol Biol (Mosk), 1988 Sep-Oct, 22(5), 1257 - 64
{Mutations in the alpha-amylase gene of Bacillus amyloliquefaciens, leading to a decrease in the temperature of protein inactivation}; Smirnova NA et al.; Alpha-amylase genes of Bacillus amyloliquefaciens, coding proteins with reduced thermostability, had been obtained as a result of hydroxylamine mutagenesis . Temperature, pH and starch concentration dependences of two mutant alpha-amylases were investigated . The synthesis of the alpha-amylases by several B . subtilis strains with different levels of extracellular proteases was also studied . The mutation containing fragments were localized and the structures of the mutations were determined . It was found that the decrease of thermostability of mutant No 141 was due to Asp to Asn change at the position No 194 of the mature protein, and for mutant No 191--due to Glu to Lys change at the position No 185.

J Gen Microbiol, 1988 Sep, 134 ( Pt 9), 2551 - 8
Comparative toxicity of Bacillus thuringiensis var . israelensis crystal proteins in vivo and in vitro; Chilcott CN et al.; Bacillus thuringiensis var . israelensis crystal proteins were purified by FPLC on a Mono Q column to yield 130, 65, 28, 53, 30-35 and 25 kDa proteins . All the purified proteins killed Aedes aegypti larvae after citrate precipitation, but the 65 kDa protein was the most toxic . A precipitated mixture of 27 and 130 kDa proteins was almost as toxic as solubilized crystals . In assays against a range of insect cell lines, the activated form (25 kDa) of the 27 kDa protein was generally cytotoxic with the lowest LC50 values in vitro . By contrast, the activated forms of the 130 kDa and 65 kDa protoxins (53 kDa and 30-35 kDa proteins, respectively) were much more specific than the 25 kDa protein in their action on dipteran cells, and each showed a unique toxicity profile which, in the case of the 130 kDa preparation, was restricted to Anopheles and Culex cell lines.

Int J Epidemiol, 1988 Sep, 17(3), 618 - 24
Secular trends of infectious disease mortality in The Netherlands, 1911-1978: quantitative estimates of changes coinciding with the introduction of antibiotics; Mackenbach JP et al.; Secular trends of mortality from 21 infectious diseases in the Netherlands were studied by inspection of age/sex-standardized mortality curves and by log-linear regression analysis . An attempt was made to obtain quantitative estimates for changes coinciding with the introduction of antibiotics . Two possible types of effect were considered: a sharp reduction of mortality at the moment of the introduction of antibiotics, and a longer lasting (acceleration of) mortality decline after the introduction . Changes resembling the first type of effect were possibly present for many infectious diseases, but were difficult to measure exactly, due to late effects on mortality of World War II . Changes resembling the second type of effect were present in 16 infectious diseases and were sometimes quite large . For example, estimated differences in per cent per annum mortality change were 10% or larger for puerperal fever, scarlet fever, rheumatic fever, erysipelas, otitis media, tuberculosis, and bacillary dysentery . No acceleration of mortality decline after the introduction of antibiotics was present in mortality from 'all other diseases' . Although the exact contribution of antibiotics to the observed changes cannot be inferred from this time trend analysis, the quantitative estimates of the changes show that even a partial contribution would represent a substantial effect of antibiotics on mortality from infectious diseases in the Netherlands.

J Am Mosq Control Assoc, 1988 Sep, 4(3), 252 - 5
Efficacy of two formulations of Bacillus thuringiensis var . israelensis (H-14) against Aedes vexans and safety to non-target macroinvertebrates; Gharib AH et al.; An experimental Sandoz formulation of Bacillus thuringiensis var . israelensis (SAN 402 SC 98) was several times more effective than Abbott ABG 6188 against larvae of Aedes aegypti and Ae . vexans in the laboratory . Field applications of SAN 402 SC 98 at 0.25 liter/ha and ABG 6188 at 1.00 liter/ha resulted in more than 97% control of Ae . vexans larvae after 48 hours, with residual activity of 24 hours or less . The amphipod, Hyallela azteca, and 4 species of water beetles were apparently unaffected by 100 ppm SAN 402 SC 98, which is 1,449 times the 24 hour LC50 for third instar Ae . vexans larvae.

Appl Environ Microbiol, 1988 Sep, 54(9), 2320 - 1
Toxicity of Bacillus sphaericus crystal toxin to adult mosquitoes; Stray JE et al.; Adult Culex quinquefasciatus mosquitoes were killed by alkaline-solubilized Bacillus sphaericus toxin when it was introduced by enema into the midgut of the insect but not when it was administered orally . Adult Aedes aegypti mosquitoes were not affected by the toxin.

J Dairy Sci, 1988 Sep, 71(9), 2384 - 7
Effect of disc moistening method on Bacillus stearothermophilus disc assay zone diameters; Ryan JJ et al.; Effect of disc moistening method on the Bacillus stearothermophilus disc assay zone size was studied . Pasteurized homogenized milk was subdivided and three levels of US Pharmacopeial reference standard penicillin G were added . Disc assay filter paper discs were moistened using the standard capillary action procedure and a method incorporating the use of a 90-microliters micropipetter . Amount of milk sample absorbed by discs prepared by the capillary action method was correlated with disc mass . Variation in amount of milk absorbed by discs was significantly greater when discs were prepared by capillary action . Zone diameter means and standard errors across all penicillin G concentrations tested were not significantly different for the two methods . The 90-microliters micropipetter method is recommended as an alternative to the capillary action method.

Oral Surg Oral Med Oral Pathol, 1988 Sep, 66(3), 287 - 9
Metronidazole in the management of anaerobic neck infection in acute leukemia; Barrett AP; A case involving a rapidly progressive neck infection in a severely neutropenic patient with acute leukemia is reported . Resolution followed the addition of metronidazole to an existing standard piperacillin/gentamicin combination, which was used primarily to cover potential gram-negative bacillary pathogens . This indicated the importance of considering extension of anaerobic cover in such infections.

Biochem J, 1988 Sep 1, 254(2), 437 - 42
Models of proteolysis of oligomeric enzymes and their applications to the trypsinolysis of citrate synthases; Else AJ et al.; A simple statistical approach was used to generate predictive models of the proteolysis of multisubunit enzymes in order to correlate the loss of enzyme activity with the loss of native subunit . The models were applied to the trypsinolysis of the citrate synthases of pig heart, Bacillus megaterium and Escherichia coli . With the dimeric citrate synthases (pig heart and B . megaterium) trypsinolysis of one of the subunits appears to destroy the activity of the whole enzymic molecule . The hexameric E . coli citrate synthase behaves like a trimer of dimeric units, each of the dimers behaving similarly to the B . megaterium and pig heart enzymes . Palmitoyl-CoA is required for the trypsinolysis of pig heart citrate synthase, and at relatively high concentrations of this compound trypsinolysis of one subunit leaves the other subunit fully active . Palmitoyl-CoA is not required for the trypsinolysis of the other citrate synthases, and high concentrations of this metabolite do not affect the correlation of proteolysis with inactivation of these enzymes.

J Immunol, 1988 Sep 1, 141(5), 1701 - 8
Human phagocytic cell responses to Mycobacterium leprae and Mycobacterium bovis Bacillus Calmette-Guérin . An in vitro comparison of leprosy vaccine components; Holzer TJ et al.; Components of current vaccines for Hansen's disease include Mycobacterium bovis Bacillus Calmette-Guerin (BCG) and killed Mycobacterium leprae . BCG infections in humans are rare and most often occur in immune-compromised individuals . M . leprae on the other hand, although not causing clinical disease in most exposed individuals, is capable of infecting and replicating within mononuclear phagocytes . Lymphocytes from patients with the lepromatous form of Hansen's disease exhibit defective lymphokine production when challenged in vitro with M . leprae . This may result in inefficient mononuclear phagocyte activation for oxidative killing . To study the ability of normal phagocytes to ingest and respond oxidatively to BCG and M . leprae, we measured phagocytic cell O2- release and fluorescent oxidative product formation and visually confirmed the ingestion of the organisms . BCG stimulated a vigorous O2- generation in neutrophils and monocytes and flow cytometric oxidative product generation by neutrophils occurred in the majority of cells . M . leprae, stimulated a weak but significant O2- release requiring a high concentration of organisms and long exposure . By flow cytometric analysis, most neutrophils were able to respond to both organisms with the generation of fluorescent oxidative products . Neutrophil oxidative responses to M . leprae were substantially less than responses seen from neutrophils exposed to BCG . By microscopic examination of neutrophils phagocytizing FITC-labeled bacteria, it was shown that both M . leprae and BCG were slowly ingested but that more BCG appeared to be associated with the cell membrane of more of the cells . When phagocytic cells were incubated with BCG and M . leprae for 30 min and subsequently examined by electron microscopy, few organisms were seen in either neutrophils or monocytes . This suggests that BCG are easily recognized and slowly ingested by normal phagocytic cells, the majority of which respond with a strong oxidative burst . M . leprae appeared to only weakly stimulate phagocyte oxidative responses and were also slowly phagocytized.

J Biochem (Tokyo), 1988 Sep, 104(3), 416 - 20
Specificity of alkaline elastase Bacillus on the oxidized insulin A- and B-chains; Tsai YC et al.; The substrate specificity of alkaline elastase Bacillus from alkalophilic Bacillus sp . Ya-B was investigated using oxidized insulin A- and B-chains . Under time-limited cleavage, the initial cleavage site of the enzyme on the oxidized insulin A-chain and B-chain was at the leucine13-tyrosine14 bond and the leucine15-tyrosine16 bond, respectively . When the cleavage was completed, three major cleavage sites and three minor cleavage sites on the A-chain, and five major cleavage sites and four minor cleavage sites on the B-chain were found . However, most of the peptides produced after complete hydrolysis of the A- or B-chain by the enzyme were composed of four to six amino acid residues . The results suggest that this enzyme cleaves the oxidized insulin A- and B-chains in a block-cutting manner.

Mol Gen Genet, 1988 Sep, 214(1), 55 - 61
Synthesis and processing of Escherichia coli TEM-beta-lactamase and Bacillus licheniformis alpha-amylase in E . coli: the role of signal peptidase I; van Dijl JM et al.; A mutant of Escherichia coli, in which signal peptidase I synthesis can be regulated, was constructed . The mutant was used to study the effects of signal peptidase I limitation on the synthesis and efficiency of processing of two proteins: the periplasmic E . coli TEM-beta-lactamase and Bacillus licheniformis alpha-amylase, which also accumulates in the periplasm of E . coli . Signal peptidase I limitation resulted in reduced rates of processing of pre-beta-lactamase and in strong inhibition of synthesis of alpha-amylase . The data suggest that beta-lactamase is processed post-translationally and that an intimate relationship exists between the synthesis and processing of alpha-amylase.

Circ Shock, 1988 Sep, 26(1), 1 - 14
Lethal shock in partially hepatectomized rats administered tumor necrosis serum; Fukushima H et al.; A minute dose of bacterial endotoxin is known to cause lethal shock in BCG (Bacillus Calmette Guerin)-sensitized mice and rats . To gain insight into the mechanism of this hypersensitivity to endotoxin, serum (tumor necrosis serum: TNS) was prepared from BCG-sensitized Lewis rats following endotoxin challenge and injected intravenously into Lewis rats 2 days after their partial hepatectomy . TNS injection caused lethal shock in Hpx (partially hepatectomized) rats but not in normally fed, fasted, or sham-operated rats . Hpx rats survived injection of serum prepared by either BCG sensitization or endotoxin challenge alone . Biochemical and histological examination of the Hpx rats injected with TNS indicated that profound hypoglycemia, hepatic and renal injury, and dysfunction of the coagulation system accompanied by hemorrhage were involved in the lethal shock . These experiments also suggested that serum component(s), probably monokine(s) derived from activated macrophages, might participate in the endotoxin shock in BCG-sensitized rats.

Rev Infect Dis, 1988 Sep-Oct, 10(5), 922 - 9
Actinobacillus actinomycetemcomitans prosthetic valve endocarditis; Grace CJ et al.; Actinobacillus actinomycetemcomitans, a fastidious gram-negative bacillus, has been reported as the cause of prosthetic valve endocarditis in 11 patients . Two additional patients are reported and the literature is reviewed . All cases occurred greater than 1 year after implantation of the prosthesis . Six of the 13 patients had had recent dental work or had poor dentition . Three patients had received endocarditis prophylaxis . Ten of 13 were cured with antibiotics alone . Only one patient suffered from congestive heart failure, and only one had documented evidence of major systemic emboli during antimicrobial therapy . Valve replacement was necessary in only two during antimicrobial therapy . A actinomycetemcomitans should be considered as a possible etiologic agent in late prosthetic valve endocarditis, particularly when blood cultures are initially negative . A regimen of a beta-lactam antibiotic in combination with an aminoglycoside is recommended for 4-6 weeks . The excellent in vitro activity of the third-generation cephalosporins and rifampin promise new therapeutic options.

Am Rev Respir Dis, 1988 Sep, 138(3), 572 - 7
Cellular influx and activation increase macrophage cytotoxicity and interleukin-1 elaboration during pulmonary inflammation in rats; Fuchs HJ et al.; This study was undertaken to investigate the effects of pulmonary inflammation induced by bacillus Calmette-Guerin (BCG) on the density distribution of lavaged alveolar macrophages . We sought to determine macrophage cytotoxicity and interleukin-1 elaboration in density-defined subpopulations of macrophages during tissue inflammation . At all time points after intravenously administered BCG, lavaged alveolar macrophages contained increased percentages of higher density cells . Alveolar macrophage cytotoxicity against the rat sarcoma cell line XC increased maximally 2 to 6 days after intravenous administration of BCG before declining on Day 13 . Macrophage interleukin-1 elaboration increased maximally 14 days after administration of BCG before declining on Day 23 . Additionally, macrophage cytotoxicity and interleukin-1 elaboration were increased above normal in cells from each of five density fractions . We conclude that a subpopulation of higher density macrophages, probably recently derived from blood monocytes, accumulates in inflammatory sites . Cellular activation increases the cytotoxicity and interleukin-1 elaboration by macrophages in all density-defined subpopulations and obscures the relationship between cellular density and function that is present in normal animals.

Mol Gen Genet, 1988 Sep, 214(1), 42 - 7
Specificity of action on mosquito larvae of Bacillus thuringiensis israelensis toxins encoded by two different genes; Delecluse A et al.; A 135 kDa protein gene and two open reading frames (ORF1 and ORF2) have been cloned from a large plasmid of Bacillus thuringiensis israelensis (Bourgouin et al . 1986) . The Escherichia coli recombinant clones containing these genes were highly toxic to larvae of Aedes aegypti, Anopheles stephensi and Culex pipiens . From subcloning experiments it was deduced that the 135 kDa polypeptide alone was responsible for the toxic activity on both A . aegypti and An . stephensi larvae . In contrast, the presence of two polypeptides, the 135 kDa protein and the ORF1 product was required for toxicity to C . pipiens larvae . The minimal toxic fragment of the 135 kDa polypeptide has been delineated . The results indicate that a polypeptide of about 65 kDa, corresponding to an amino-terminal part of the 135 kDa protein is sufficient for toxicity . Sequence comparisons indicate that the ORF1 product may correspond to an N-terminal part of a rearranged 130 kDa protein.

J Gen Microbiol, 1988 Sep, 134 ( Pt 9), 2559 - 65
Protein phosphorylation in Bacillus thuringiensis during growth and delta-endotoxin production; Watson GM et al.; At least 14 phosphopolypeptides in which the phosphate groups were present as mono-esters were detected by pulse labelling of Bacillus thuringiensis subsp . kurstaki HD-1-Dipel with {32P}orthophosphate at different stages of growth and differentiation . Marked changes in the profile of phosphopolypeptides were observed primarily during the late exponential phase of growth . Several phosphopolypeptides co-purified with the endotoxin crystal of this subspecies and the phosphoamino acid residue of the most abundant (Mr 25,000) phosphopolypeptide was identified as phosphothreonine . Comparison of the phosphopolypeptides in endotoxin crystals from several subspecies suggested that Mr 25,000 species might be a common component.

Mol Cell Biol, 1988 Sep, 8(9), 3636 - 46
Structure and regulation of a nuclear gene in Saccharomyces cerevisiae that specifies MRP7, a protein of the large subunit of the mitochondrial ribosome; Fearon K et al.; The gene for MRP7, a 40-kilodalton protein of the large subunit of the yeast mitochondrial ribosome, was identified in a lambda gt11 expression library by immunological screening with a monoclonal antibody to MRP7 . An intact copy of MRP7 was then isolated from a yeast genomic library by colony hybridization . Gene disruption showed that MRP7 protein was essential for ribosomal function . Sequencing of MRP7 revealed a coding region for a basic (pI 10.6), 43.2-kilodalton protein containing 371 amino acid residues . Amino acid residues 28 to 112 of the deduced MRP7 sequence aligned with the 84 residues of the Escherichia coli ribosomal protein L27, but no significant similarity was detected between the carboxy-terminal 259 amino acids of MRP7 and other protein sequences in existing computer data bases . Within the aligned region, there was 49% amino acid identity between MRP7 and L27, compared with the 57% identity observed between L27 and its homolog in Bacillus stearothermophilus . The steady-state levels of the MRP7 protein and its mRNA were monitored in response to catabolite repression and to increased dosage of the MRP7 gene . The response to catabolite repression was characterized by a ninefold change in the level of the protein and little, if any, change in the level of the mRNA . In cells carrying the MRP7 gene on a high-copy-number plasmid, the mRNA was increased 20-fold, but there was no significant increase in MRP7 protein . Furthermore, MRP7 mRNA and protein accumulated at normal levels in {rho0} cells, which are devoid of 21S rRNA, indicating that the protein is relatively stable in the absence of ribosome assembly . Together, these results suggest that MRP7 is regulated posttranscriptionally, probably at the level of protein synthesis rather than protein turnover.

Proc Natl Acad Sci U S A, 1988 Sep, 85(18), 6987 - 91
Lysogeny and transformation in mycobacteria: stable expression of foreign genes; Snapper SB et al.; Requisite to a detailed understanding of the molecular basis of bacterial pathogenesis is a genetic system that allows for the transfer, mutation, and expression of specific genes . Because of the continuing importance of tuberculosis and leprosy worldwide, we initiated studies to develop a genetic system in mycobacteria and here report the use of two complementary strategies to introduce and express selectable genetic markers . First, an Escherichia coli cosmid was inserted into the temperate mycobacteriophage L1, generating shuttle phasmids replicating as plasmids in E . coli and phage capable of lysogenizing the mycobacterial host . These temperate shuttle phasmids form turbid plaques on Mycobacterium smegmatis and, upon lysogenization, confer resistance to superinfection and integrate within the mycobacterial chromosome . When an L1 shuttle phasmid containing a cloned gene conferring kanamycin resistance in E . coli was introduced into M . smegmatis, stable kanamycin-resistant colonies--i.e., lysogens--were obtained . Second, to develop a plasmid transformation system in mycobacteria, M . fortuitum/E . coli hybrid plasmids containing mycobacterial and E . coli replicons and a kanamycin-resistance gene were constructed . When introduced into M . smegmatis or BCG (Mycobacterium tuberculosis typus bovinus var . Bacille-Calmette-Guerin) by electroporation, these shuttle plasmids conferred stable kanamycin resistance upon transformants . These systems should facilitate genetic analyses of mycobacterial pathogenesis and the development of recombinant mycobacterial vaccines.

Mol Biol (Mosk), 1988 Sep-Oct, 22(5), 1265 - 71
{Two-codon mutagenesis of alpha-amylase gene of Bacillus amyloliquefaciens}; Smirnova NA et al.; The oligonucleotide encoding Bam HI recognition site having the structure pCGGGATC had been inserted into the recognition sites MspI of the B . amyloliquefaciens alpha-amylase gene, which was cloned in pTG29B plasmid . The alpha-amylase gene had no BamHI sites before mutagenesis . The set of pNSBamHI plasmids with BamHI site at four different positions was obtained . It was shown that all the mutant alpha-amylases possess different specific activities . One of the mutant proteins possesses reduced thermostability . The mutant alpha-amylases can be used for further experiments on protein-engineering of liquefying-type alpha-amylases.

Antibiot Khimioter, 1988 Sep, 33(9), 661 - 5
{Effect of salts on the lytic activity of gramicidin S and its derivatives}; Bulgakova VG et al.; Potassium and sodium chlorides, sulfates, acetates and phosphates activated the lytic action of gramicidin S and its derivatives on protoplasts of M . lysodeikticus . The derivatives used were positively charged and neutral by the free amino groups in the ornithine moieties . The salts had no effect on lysis of the bacillar protoplasts by gramicidin S and its positively charged derivatives . The lytic effect of the neutral derivative on the bacillar protoplasts markedly increased in the presence of the salts, activation of the lysis by the phosphates being more pronounced than that by the other salts . Increased membrane activity of gramicidin S in the presence of the salts was not connected with association of the substance molecules in solution . Probably it was due to increased destruction of the membranes at the account of activated detergent effect of the antibiotic and its derivatives.

J Chromatogr, 1988 Aug 31, 448(1), 109 - 16
Modified general affinity adsorbent for large-scale purification of penicillinases; Kiss L et al.; N-Acetyl-D-(-)-penicillamine as a stable second-generation biospecific affinity ligand has previously been suggested for purification of Bacillus cereus 569/H beta-lactamase I . A complex spacer arm is coupled with the matrix by using epichlorohydrin and phloroglucinol doubly activated with divinyl sulphone in the meta position . Coupling of D-(-)-penicillamine ligand resulted in an active affigel . However, we found that two affinity ligands in close proximity prevents simultaneous binding of two penicillinase molecules, therefore one ligand is superfluous . Our results show that: (1) shortening the spacer arm by direct activation of the matrix with divinyl sulphone is satisfactory to produce the affinity material with N-acetyl-D-(-)-penicillamine; (2) incorporation of 15 mumol of N-acetyl-D-(-)-penicillamine per ml of wet Sepharose 4B satisfies the maximum binding capacity requirements of the affigel (about half of the originally incorporated amount of ligand); (3) our simplified affinity adsorbent is generally applicable for large-scale purification of penicillinases to homogeneity from various bacterial sources by the convenient batch method without prior concentration of these enzymes; (4) reacetylation for four/five times can regenerate the original binding capacity of the affigel.

J Biol Chem, 1988 Aug 25, 263(24), 11800 - 1
Crystallization and preliminary X-ray diffraction studies of a toxic crystal protein from a subspecies of Bacillus thuringiensis; Garfield JL et al.; The toxic crystal protein (Mr 64,000) from a subspecies of the bacterium Bacillus thuringiensis has been solubilized and recrystallized yielding diffraction quality crystals . Crystals are obtained by a change in pH and ionic strength using Na2CO3 . They can also be obtained by a change in ionic strength only using NaBr as the precipitant . The space group of both forms is C222(1) with a = 133, b = 116, c = 104 A and one molecule/asymmetric unit . Still photographs show reflections to 3.0-A resolution.

J Biol Chem, 1988 Aug 25, 263(24), 11814 - 9
Analogs of diaminopimelic acid as inhibitors of meso-diaminopimelate dehydrogenase and LL-diaminopimelate epimerase; Lam LK et al.; Analogs 1-8 of diaminopimelic acid (DAP) were synthesized and tested for inhibition of purified meso-DAP D-dehydrogenase from Bacillus sphaericus and of LL-DAP epimerase from Escherichia coli . The dehydrogenase was assayed by monitoring NADPH formation spectrophotometrically at 340 nm . N-Hydroxy DAP 4, N-amino DAP 5, and 4-methylene DAP 6 are substrates of the dehydrogenase with relative rates exceeding those of the meso isomers of the thia analogs 1ab, 2ab, and 3ab . DAP epimerase was assayed by coupling the epimerization of LL-DAP to DL-DAP (Km = 0.26 mM) with the dehydrogenase-catalyzed oxidation of DL-DAP by NADP . Lanthionine isomers 1ab and 1c were stronger inhibitors of the epimerase (Ki = 0.18 mM, Ki' = 0.67 mM, and Ki = 0.42 mM, respectively) than the corresponding meso-sulfoxide 2ab or the meso-sulfone 3ab . Other isomers of 2 and 3, as well as compounds 7 and 8, showed no epimerase inhibition . N-Hydroxy DAP 4 was the most potent competitive inhibitor (Ki = 0.0056 mM) of the epimerase, whereas N-amino DAP 5 is weaker (Ki = 2.9 mM) and 4-methylene DAP 6 is a noncompetitive inhibitor (Ki' = 0.95 mM) . Although none of the analogs tested showed time-dependent inactivation of either enzyme, compounds 4, 5, 6, and 7 display substantial antibacterial activities . Possible mechanisms of epimerase inhibition and significance of the DAP pathway as a target for antibiotics are discussed.

J Mol Biol, 1988 Aug 20, 202(4), 913 - 5
Barnase and barstar . Expression of its cloned inhibitor permits expression of a cloned ribonuclease; Hartley RW; Barnase is the extracellular ribonuclease of Bacillus amyloliquefaciens and barstar its specific intracellular inhibitor . The gene for barstar has now been cloned and sequenced . When the wild-type gene for barnase is reconstructed from its previously cloned parts on the same plasmid as the barstar gene, the lethal effect of its expression is suppressed . A plasmid has been devised which directs the secretion of 100 mg per active barnase liter by Escherichia coli and another which provides large (500 to 1000 mg/l) yields of barstar . The structure of these plasmids and the derived 89 amino acid sequence of barstar are reported.

J Immunol, 1988 Aug 15, 141(4), 1083 - 90
High strength binding of P815 mastocytoma cells is not necessary for their lysis by macrophages which have been primed and triggered in vitro; Lu CY et al.; We have examined the hypothesis that binding of P815 mastocytoma cells is a necessary step in lysis of these cells by macrophages which are both "primed" and "triggered" in vitro, Macrophages "primed" by conditioned media containing IFN-gamma, or by rIFN-gamma have an increased ability to bind P815 . However, adding either heat-killed Listeria or endotoxin to trigger the primed macrophages has opposite effects on lysis and binding of P815 . Lysis is increased . Binding is dramatically decreased . This is true when centrifugal forces of 200 x g, 400 x g, and 800 x g are used to disrupt P815-macrophage binding . Although 100% of P815 cells bound by cytotoxic macrophages are lysed, a large additional population of unbound P815 is also lysed . Detailed kinetic studies indicate that macrophages do not rapidly bind and lyse several cycles of P815 . There is an initial lag period of 4 to 6 h before P815 lysis can be detected, and completion of lytic events then occurs within 12 to 14 h . Lysis of P815 bound to cytotoxic macrophages is slightly slower than lysis of the total population of bound and unbound P815 . In contrast, D3.1, a cloned CD4+ T cell line, is tightly bound to macrophages but not lysed efficiently . When macrophages are simultaneously confronted with P815 and macrophage-bound D3.1, only the former are lysed . Altogether, the data indicate that P815-macrophage binding, as operationally defined by our assay, is not a necessary step for lysis . These results, by use of macrophages primed and triggered in vitro, are in contrast to previously reported experiments examining P815 binding and lysis by macrophages activated in vivo by infection with bacillus Calmette-Guerin.

FEBS Lett, 1988 Aug 15, 236(1), 251 - 5
Different phosphorylated forms of an insulin-sensitive glycosylphosphatidylinositol from rat hepatocytes; Merida I et al.; Labeling with {3H}galactose was employed to isolate a glycosylphosphatidylinositol from rat hepatocytes which might be involved in the action of insulin . The polar head group of this glycosylphosphatidylinositol was generated by phosphodiesterase hydrolysis with a phosphatidylinositol-specific phospholipase C from Bacillus cereus . By Dowex AG1 x 8 chromatography the polar head group could be separated into three radioactive peaks eluting at 100 mM (peak I), 200 mM (peak II) and 500 mM (peak III) ammonium formate, respectively . Peak III was the most active as an inhibitor of the cAMP-dependent protein kinase . Treatment of peak III with alkaline phosphatase markedly reduced its activity on cAMP-dependent protein kinase . When peaks I, II or III were treated with alkaline phosphatase and analyzed again by Dowex AG1 x 8 chromatography, the radioactivity eluted with the aqueous fraction . The above results indicate that the polar head group of the insulin-sensitive glycosylphosphatidylinositol from rat hepatocytes exists in three different phosphorylated forms and that the biological activity of this molecule depends on its phosphorylation state.

Am J Infect Control, 1988 Aug, 16(4), 147 - 51
A creative method for determining the immunization status of a community hospital medical staff; Scheckler WE; During 1985 and 1986, 191 of the 212 (90%) medical staff members in active private practice, who were surveyed responded to a one-page report regarding the status of their immunity and immunization to important infectious illnesses . Response rates were improved by including the survey form with the hospital privilege-renewal form that was required to be returned to the medical staff office . Of the respondents 96% reported prior history of chickenpox and 95% reported prior immunization to tetanus; of the 42 who reported prior bacille Calmette-Guerin (BCG) vaccination, only 17 reported a positive tuberculin skin test . An additional 32 medical staff members had a history of a positive tuberculin skin test with no prior BCG vaccination . Only 2 (1%) reported prior history of hepatitis B, and only 15 (8%) reported hepatitis B vaccination begun or completed at the time of the survey . This survey allows targeting of infection control activities, such as implementation of the hepatitis B vaccine promotion program and serologic testing of those with a negative history of chickenpox, to be developed for the medical staff . It allows the hospital infection control team instant access to data that can be simply obtained and updated regularly when information is needed about the immunization status of selected medical staff members during an outbreak situation.

Appl Environ Microbiol, 1988 Aug, 54(8), 2134 - 5
Resistance of some common fungi to gamma irradiation; Saleh YG et al.; Ten species of fungi representing the genera Alternaria, Aspergillus, Caldosporium, Curvularia, Fusarium, and Penicillium were examined for their relative resistance to gamma irradiation from a 137Cs source . Inactivation doses for dematiaceous fungi in agar medium ranged from 0.6 to greater than 1.7 megarads, whereas those for moniliaceous fungi were less than 0.3 megarad . D10 values (the dose required to reduce the inoculum by 1 log) for Curvularia geniculata (greater than 0.29 megarad) exceeded those for control spores of Bacillus pumilus (0.15 megarad).

J Dairy Sci, 1988 Aug, 71(8), 2292 - 6
Persistence of residues in milk following antibiotic treatment of dairy cattle; Seymour EH et al.; A study was conducted to determine the persistence of antibiotic residues in milk beyond the recommended withdrawal period . Composite milk samples (n = 122) were collected from 58 lactating dairy cows in the university herd receiving antibiotic treatments for any reason but only when administered as a single drug . Samples were obtained 72 h posttreatment and sampling continued every 24 h until result for antibiotic residue was negative by the Bacillus stearothermophilus disc assay . The antibiotic (n = 7) administered accounted for significant variation in drug persistence; however, route of administration, case number (for cows treated for more than one episode), number of days treated, animal's body weight, lactation number, and daily milk production did not affect drug persistence . Chi-square analysis indicated that 21% of milk samples were positive for residues beyond the recommended withholding period . Milk samples from cows treated with cephapirin and penicillin were the only samples that exceeded recommended withdrawal times . Often doses administered exceeded label directions.

Biochimie, 1988 Aug, 70(8), 1013 - 8
Mechanisms of Mycobacterium leprae-specific T-cell deficiency in lepromatous leprosy; Bach MA et al.; Patients suffering from lepromatous leprosy fail to develop an efficient cell-mediated immunity towards Mycobacterium leprae, the causative agent . The mechanism of such a specific T-cell tolerance to the bacillus remains a key question in the pathophysiology of leprosy . Macrophages do not show any intrinsic defect in phagocytizing and killing M . leprae or in presenting antigen to helper T-cells . On the other hand, M . leprae-reactive helper T-cells do persist in lepromatous patients, but their activation appears to prevented by active suppressor mechanisms, involving both suppressor T-cells and macrophages . The target of this specific suppression could be the interleukin 2-producing T-cell subset . A better molecular definition of M . leprae antigens, both by monoclonal antibodies and T-cell clones, should open new perspectives for further analysis of the regulation of immune responses to M . leprae.

Pediatr Infect Dis J, 1988 Aug, 7(8), 578 - 81
Booster effect of tuberculin testing in healthy 6-year-old school children vaccinated with Bacillus Calmette-Guérin at birth in Santiago, Chile; Sepulveda RL et al.; In order to determine whether tuberculin testing caused a booster effect in children vaccinated with Bacillus Calmette-Guerin (BCG) at birth, we studied forty 6-year-olds by repeat tuberculin testing 2 weeks later on the contralateral forearm . All children were healthy and had no known exposure to tuberculosis . None of the children had a history of mycobacteriosis other than tuberculosis . The mean induration was 2.3 +/- 1.8 mm for the first tuberculin reaction and 7.6 +/- 3.3 mm for the second tuberculin reaction (P less than 0.005) . Four children had positive reactions (greater than or equal to 10 mm) to the first purified protein derivative test; 18 children were positive upon retesting . Eleven of these latter children had increases of at least 6 mm from reactions less than 10 mm to greater than or equal to 10 mm . The size of the BCG scar was significantly correlated to the size of both the first and second purified protein derivative reactions (P less than 0.01), suggesting that the increased reactivity upon retesting was a consequence of sensitization induced by BCG vaccination 6 years earlier . All children remained healthy after this study was completed . Retesting of tuberculin reactivity within 2 weeks in BCG-vaccinated children with reactions less than 10 mm will produce reactions greater than 10 mm in some healthy children who may not require antituberculosis treatment.

J Clin Microbiol, 1988 Aug, 26(8), 1571 - 4
Plasmid, serotypic, and enterotoxin analysis of Bacillus cereus in an outbreak setting; DeBuono BA et al.; Bacillus cereus is a recognized agent of food-borne disease . In this report we describe an outbreak of B . cereus gastroenteritis associated with consumption of beef stew among patients and staff at a Rhode Island nursing home . The beef had been improperly stored after preparation . The predominant symptoms of the illness were cramps and diarrhea; it lasted an average of 16 h . No deaths occurred . The organism was recovered from 10 of 23 stools collected from ill patients and 1 of 21 stools collected from controls (P = 0.0044, Fisher's two-tailed exact test) . All isolates had the same biotype and serotype, newly designated H.26; all elaborated the diarrheal B . cereus enterotoxin when tested in rabbits by the vascular permeability reaction; and all had identical plasmid profiles, which differed from those of B . cereus strains selected randomly from other outbreaks . Plasmid analysis may prove to be a useful new tool in investigating outbreaks of B . cereus food poisoning.

Proc Natl Acad Sci U S A, 1988 Aug, 85(15), 5678 - 82
Induction of protective immunity against Schistosoma mansoni by vaccination with schistosome paramyosin (Sm97), a nonsurface parasite antigen; Pearce EJ et al.; Paramyosin (Sm97), a 97-kDa myofibrillar protein identified by the unusually monospecific antibody response induced by intradermal vaccination of mice with a complex soluble worm antigen preparation (SWAP) of adult Schistosoma mansoni administered with bacillus Calmette-Guerin (BCG), was purified and tested for its capacity to protect mice against challenge infection . When administered intradermally with BCG at total doses of only 4-40 micrograms per mouse, both the native molecule and a recombinant expression product containing approximately 50% of the whole protein were found to confer significant resistance (26-33%) against challenge infection, while 2 mg of unfractionated SWAP was required to induce similar levels of protection . In addition, paramyosin was shown to stimulate T lymphocytes from vaccinated mice to produce lymphokines {e.g., gamma interferon (IFN-gamma)} that activate macrophages to kill schistosomula . Neither schistosome myosin nor a heterologous paramyosin from a different invertebrate genus were protective, indicating a requirement for specific epitopes in the immunization . That the protection induced by paramyosin involves a T-cell-mediated mechanism was supported by the failure of anti-paramyosin antibodies to passively transfer significant resistance to infection to recipient mice . Lymphocytes from mice vaccinated with paramyosin were found to produce IFN-gamma in response to living schistosomula, suggesting that during challenge infection of vaccinated hosts, paramyosin (a nonsurface antigen) may elicit a protective T-cell response as a consequence of its release from migrating parasite larvae . Paramyosin-depleted SWAP was also found to be protective as well as stimulatory for T lymphocytes from SWAP-vaccinated mice, indicating that other antigens in this preparation may have immunoprophylactic potential . In summary, these results (i) suggest that the induction of T-cell-dependent cell-mediated immunity against soluble nonsurface antigens may be an effective strategy for immunization against multicellular parasites and (ii) in the case of schistosomes, identify paramyosin as a candidate vaccine immunogen in this category.

Am J Trop Med Hyg, 1988 Aug, 39(2), 173 - 8
Analysis and preparation of Bartonella bacilliformis antigens; Knobloch J; Twenty-four antigens of Bartonella bacilliformis, a bacterium which causes bartonellosis in residents of high altitude valleys of the Andes, were identified by immunoblot and immunoprecipitation using rabbit anti-Bartonella sera as well as sera of patients . The antigens were designated according to their relative molecular mass which ranged from 16 to 160 kDa . Twelve antigens were detected by antibodies in sera of bartonellosis patients using immunoblot, of which six antigens were detected by immunoprecipitation . Antigens 25, 46, 65, 75, 99, and 160 were identified as probable cell wall antigens . Antigens 50, 65, and 75 detected long-persisting antibodies . Crude Bartonella antigen applied to ELISA reacted with anti-Chlamydia psittaci antibody as well as with antibody of unknown identity in human sera, whereas immunoblot and immunoprecipitation with Triton soluble antigens revealed Bartonella-specific results . Seven Bartonella antigens were prepared by high performance liquid chromatography of which one antigen (48 kDa) reacted Bartonella-specific when applied to ELISA . It was concluded that specificity of antibody determination with crude Bartonella antigen should be confirmed by either immunoblot or immunoprecipitation.

Antibiot Khimioter, 1988 Aug, 33(8), 563 - 6
{Enzyme activity of Bacillus polymyxa 153, the producer of polymyxin B, during sporulation and biosynthesis}; Ermakova GN et al.; Metabolic properties of Bacillus polymyxa 153 were studied during vegetative growth, polymyxin B biosynthesis and active sporulation . In the cell extracts there was detected activity of exoproteases, endoproteases, tricarboxylic acid cycle dehydrogenases and pyruvate dehydrogenase . The enzymes activity in the cells growing into spores was higher than that in the cells of the vegetative developmental type . The activity of the enzymes depended on the culture age.

J Bacteriol, 1988 Aug, 170(8), 3761 - 4
Transduction in Bacillus stearothermophilus; Welker NE; Temperate and virulent bacteriophages isolated from soil were shown to carry out generalized transduction of Bacillus stearothermophilus NUB36 . A transducing frequency of 1 X 10(-5) to 7 X 10(-4) was obtained for temperate phages TP-42 and TP-56 . The transducing frequency for virulent phage TP-68 was two to three orders of magnitude lower . Cotransfer analysis with the three phages showed that hom-1 is linked to thr-1 and that gly-1 is linked to his-1.

J Bacteriol, 1988 Aug, 170(8), 3575 - 83
A Bacillus thuringiensis subsp . israelensis gene encoding a 125-kilodalton larvicidal polypeptide is associated with inverted repeat sequences; Bourgouin C et al.; A gene encoding a 125-kilodalton (kDa) mosquitocidal delta-endotoxin was cloned from the 72-MDa resident plasmid of Bacillus thuringiensis subsp . israelensis . This gene is similar in its 3' region to the gene encoding the 135-kDa protein previously cloned (C . Bourgouin, A . Klier, and G . Rapoport, Mol . Gen . Genet . 205:390-397, 1986) . Escherichia coli recombinant clones harboring the 125-kDa gene were toxic to larvae of the three mosquito species Aedes aegypti, Anopheles stephensi, and Culex pipiens . In addition, the B . thuringiensis subsp . israelensis DNA fragment carrying the 125-kDa protein gene contains two sets of inverted repeat sequences, identified either by the S1 nuclease method or by electron microscopic observation . The structural organization of inverted repeat sequences and of the 125-kDa gene was analyzed and suggests that this B . thuringiensis subsp . israelensis delta-endotoxin gene is located within a transposable element.

Eur J Biochem, 1988 Aug 1, 175(2), 213 - 20
Nucleotide sequence and expression in Escherichia coli of the gene coding for sphingomyelinase of Bacillus cereus; Yamada A et al.; Bacillus cereus secretes phospholipases C, which hydrolyze phosphatidylcholine, sphingomyelin and phosphatidylinositol . A 7.5-kb HindIII fragment of B . cereus DNA cloned into Escherichia coli, with pUC18 as a vector, directed the synthesis of the sphingomyelin-hydrolyzing phospholipase C, sphingomyelinase . Nucleotide sequence analysis of the subfragment revealed that it contained two open reading frames in tandem . The upstream truncated open reading frame corresponds to the carboxy-terminal portion of the phosphatidylcholine-hydrolyzing phospholipase C, and the downstream open reading frame to the entire translational portion of the sphingomyelinase . The two phospholipase C genes form a gene cluster . As inferred from the DNA sequence, the B . cereus sphingomyelinase has a signal peptide of 27 amino acid residues and the mature enzyme comprises 306 amino acid residues, with a molecular mass of 34233 Da . The signal peptide of the enzyme was found to be functional in protein transport across the membrane of E . coli . The enzymatic properties of the sphingomyelinase synthesized in E . coli resemble those of the donor strain sphingomyelinase . The enzymatic activity toward sphingomyelin was enhanced 20-30-fold in the presence of MgCl2, and the adsorption of the enzyme onto erythrocyte membranes was accelerated in the presence of CaCl2.

Biochemistry, 1988 Jul 26, 27(15), 5525 - 30
Tyrosyl-tRNA synthetase acts as an asymmetric dimer in charging tRNA . A rationale for half-of-the-sites activity; Ward WH et al.; Tyrosyl-tRNA synthetase from Bacillus stearothermophilus is a classical example of an enzyme with half-of-the-sites activity . The enzyme crystallizes as a symmetrical dimer that is composed of identical subunits, each having a complete active site . In solution, however, tyrosyl-tRNA synthetase binds tightly, and activates rapidly, only 1 mol of Tyr/mol of dimer . It has recently been shown that the half-of-the-sites activity results from an inherent asymmetry of the enzyme . Only one subunit catalyzes formation of Tyr-AMP, and interchange of activity between subunits is not detectable over a long time scale . Paradoxically, however, the kinetics of tRNA charging are biphasic with respect to {Tyr}, suggesting that both subunits of the dimer are catalytically active . This paradox has now been resolved by kinetic analysis of heterodimeric enzymes containing different mutations in each subunit . Biphasic kinetics with unchanged values of KM for Tyr are maintained when one of the two tRNA-binding domains is removed and also when the affinity of the "inactive" site for Try is reduced by 2-58-fold . The biphasic kinetics do not result from catalysis at both active sites, but instead appear to result from two molecules of Tyr binding sequentially to the same site . A second molecule of Tyr perhaps aids the dissociation of Tyr-tRNA by displacing the tyrosyl moiety from its binding site . A monomer of the enzyme is probably too small to allow both recognition and aminoacylation of a tRNA molecule . This could explain the requirement for the enzyme to function as an asymmetric dimer.

Biochem J, 1988 Jul 15, 253(2), 533 - 9
A sulphate metabolizing centre in Euglena mitochondria; Saidha T et al.; We have previously shown that a sulphate activating system is present on the outside of the inner mitochondrial membrane of Euglena gracilis Klebs . var . bacillaris Cori, but efforts to couple this system to ATP produced from oxidative phosphorylation were unsuccessful . In the present work we show that the concentration of Pi ordinarily used to support oxidative phosphorylation in these mitochondria (10 mM) inhibits sulphate activation completely; by reducing the concentration of Pi 10-fold, both processes proceeded normally . Sulphate activation under these conditions is inhibited nearly completely by the uncouplers of oxidative phosphorylation dinitrophenol (0.1 mM) and carbonyl cyanide m-chlorophenylhydrazone (CCCP) (0.2 microM) . Sulphate reduction to form free cysteine, most of which appears outside the organelle, and in the cysteine of mitochondrial protein can be demonstrated in the same preparations, is membrane-bound and is inhibited by chloramphenicol (100 micrograms/ml), NaN3 (5 mM), KCN (100 microM); dinitrophenol (0.1 mM) or CCCP (0.2 microM) . Digitonin fractionation of the mitochondria into mitoplasts, outer membranes and an intermembrane fraction show that reduction of 35SO4(2-) to form free cysteine and cysteine of protein is located on the mitoplasts; adenosine 5'-phosphosulphate sulphotransferase, the first enzyme of sulphate reduction, is found in the same location . Sulphate activation is highly enriched in the mitochondrial fraction of Euglena; the small amount found in the chloroplast fraction can be attributed to mitochondrial contamination . Thus, in Euglena, sulphate activation and reduction are contained in a sulphate metabolizing centre on the outside of the mitochondrial inner membrane; this centre appears to supply the mitochondrion and the rest of the cell with the products of sulphate activation as well as with reduced sulphur in the form of cysteine . Mitochondria from wild-type Euglena cells and from W10BSmL, a mutant lacking plastids completely, appear to be similar in the properties studied.

J Immunol, 1988 Jul 15, 141(2), 627 - 35
Regulation of activated macrophage antimicrobial activities . Cooperation of lymphokines for induction of resistance to infection; Davis CE et al.; Macrophages treated with the soluble products of Ag-stimulated spleen cells from bacillus Calmette-Guerin-infected C3H/HeN mice (lymphokines) (LK} before infection developed the capacity to resist infection with obligately intracellular amastigotes of the protozoan parasite, Leishmania major: 40 to 60% fewer cells in LK-treated cultures were infected 2 h after exposure to parasites than cells in medium-treated controls . Macrophages treated with LK depleted of IFN-gamma failed to acquire this activated macrophage effector activity . Paradoxically, IFN-gamma by itself was also not effective . Activity of the ineffective, IFN-gamma-depleted LK was restored, however, by addition of 10 to 100 U/ml IFN-gamma, itself inactive . The induction of this antimicrobial activity was the result of the interaction of macrophages and several molecularly distinct LK, and IFN-gamma was a necessary but insufficient activation signal . The activation of macrophage resistance to infection by LK was 1) not signal sequence dependent, 2) absent in cells treated with the second signal at lower (4 degrees C) temperatures and in the presence of protein synthesis inhibitors, and 3) induced by the cooperation of IFN-gamma and LK of m.w . 45,000 and 33,000 . These factors in LK constituted more than 85% total LK activity for induction of resistance to infection . A minor activity in LK, of m.w . 20,000, could apparently induce this effector activity in the absence of IFN-gamma: this activity was less than 15% of total LK activity.

J Immunol, 1988 Jul 15, 141(2), 607 - 13
Exact definition of species-specific and cross-reactive epitopes of the 65-kilodalton protein of Mycobacterium leprae using synthetic peptides; Anderson DC et al.; With the use of solid phase synthesis of peptides corresponding to major and minor peaks in a Hopp-Woods hydrophilicity plot, the epitopes for 10 of 14 known different mAb to the Mycobacterium leprae 65-kDa protein, a prominent T and B cell Ag of this bacillus, have been located in the primary structure . Five epitopes have been precisely mapped by using the synthetic peptides in inhibition ELISA experiments, and five others have been located on peptides of 22 amino acids or less in length . The epitope of an important species-specific antibody, IIIE9, which may be useful for seriodiagnosis of leprosy, appears to be distinguished from the epitope of the antibody IVD2, widely cross-reactive among mycobacteria, not by its sequence, but only by its critical residues . All epitopes studied appear continuous insofar as can be determined by this approach.

Biochemistry, 1988 Jul 12, 27(14), 4966 - 70
Inhibition of alanine racemase by alanine phosphonate: detection of an imine linkage to pyridoxal 5'-phosphate in the enzyme-inhibitor complex by solid-state 15N nuclear magnetic resonance; Copie V et al.; Inhibition of alanine racemase from the Gram-positive bacterium Bacillus stearothermophilus by (1-aminoethyl) phosphonic acid (Ala-P) proceeds via a two-step reaction pathway in which reactivation occurs very slowly . In order to determine the mechanism of inhibition, we have recorded low-temperature, solid-state 15N NMR spectra from microcrystals of the {15N}Ala-P-enzyme complex, together with spectra of a series of model compounds that provide the requisite database for the interpretation of the 15N chemical shifts . Proton-decoupled spectra of the microcrystals exhibit a line at approximately 150 ppm, which conclusively demonstrates the presence of a protonated Ala-P-PLP aldimine and thus clarifies the structure of the enzyme-inhibitor complex . We also report the pH dependence of Ala-P binding to alanine racemase.

FEBS Lett, 1988 Jul 4, 234(1), 169 - 71
Isolation and characterization of restriction endonuclease BstYI from Bacillus stearothermophilus Y406; Chen ZF et al.; BstYI, an isoschizomer of XhoII and MflI, has been purified from Bacillus stearothermophilus Y406 . This enzyme recognized 5'...Pu/GATCPy...3' in DNA and cleaved between Pu and G in this sequence . BstYI can be easily isolated and purified by heparin-agarose column chromatography in a high yield (8000 units BstYI can be obtained per g wet wt of cells).

J Gerontol, 1988 Jul, 43(4), M97 - 100
Tuberculosis, tuberculin reactivity, and delayed cutaneous hypersensitivity in nursing home residents; Creditor MC et al.; When all residents of a 460-bed nursing home were tuberculin tested after discovery of a fatal case of pulmonary tuberculosis, 34% reacted, including 6% who gave boosted reactions . Twenty-four of 262 (9.2%) nonreactors converted to tuberculin reactors 6 months after exposure . Six of the convertors were among the 21% of the residents who were originally considered anergic on the basis of negative Candida and Trichophyton skin tests . These results confirm the observation that aged nursing home residents have lower rates of tuberculin reactivity than earlier in their lives, and that tuberculosis is a nosocomial infection in nursing homes . However, generalized immune senescence cannot be invoked as a reason for apparent susceptibility because the very marker of infection--the development of tuberculin reaction--is evidence of some degree of immune competence . Furthermore, the presence of cutaneous anergy as clinically determined does not predict inability to develop an immune response to the tubercle bacillus.

CMAJ, 1988 Jul 1, 139(1), 41 - 4
{Tuberculin reactors among refugee status claimants newly arrived in Canada}; Godue CB et al.; A large number of new cases of tuberculosis are reported among immigrants to Canada every year . For public health purposes immigration policy requires that all immigrants undergo chest radiography and that those found to be infectious receive treatment and follow-up . Since 1984 the clinique Sante-Accueil of the departement de Sante communautaire, hopital Saint-Luc, Montreal, has participated in the screening of immigrants arriving in Quebec who claim refugee status . We undertook a descriptive study to determine the prevalence of tuberculous infection in this population . Of 865 tuberculin tests administered and read, 329 (38%) produced reactions 10 mm or more in diameter . The proportion of positive reactions was higher among older immigrants and varied by country of origin, from 30% for people from El Salvador to 61% for those from Haiti . A history of vaccination with bacille Calmette-Guerin or the presence of a scar was not associated with a positive reaction . Three cases of pulmonary tuberculosis were identified, for a rate of 118/100,000 . Our findings underline the importance of systematically screening immigrants for tuberculous infection with the tuberculin test.

J Bacteriol, 1988 Jul, 170(7), 3206 - 12
Regulation of the penicillinase genes of Bacillus licheniformis: interaction of the pen repressor with its operators; Wittman V et al.; The synthesis of the inducible enzyme penicillinase of Bacillus licheniformis is negatively controlled by a repressor (D.A . Dubnau and M.R . Pollock, J . Gen . Microbiol . 41:7-21, 1965; D . J . Sherratt and J . F . Collins, J . Gen . Microbiol . 76:217-230,1973) . The molecular organization of the genes coding for penicillinase (penP) and its repressor (penI) has recently been determined (T . Himeno, T . Imanaka, and S . Aiba, J . Bacteriol . 168:1128-1132, 1986) . These two genes are transcribed divergently from within a 364-nucleotide region separating the coding sequences . We cloned and sequenced the repressor gene (penIc) from strain 749/C that constitutively produces penicillinase . The penIc and penI+ (wild-type) genes were expressed in Escherichia coli . Complementation analysis indicated that the repressor is the only trans-acting protein required to regulate the expression of the penI and penP genes . We purified the wild-type repressor protein, used it in gel retardation and DNase I protection experiments, and identified three operators positioned in the region between the penP and penI coding sequences . The spatial arrangement of the operators and the hierarchy in repressor binding seen in the protection experiments indicate that (i) the penI gene product represses the expression of the penP gene by physically blocking the RNA polymerase-binding site and (ii) the penI gene is autoregulated.

J Gen Microbiol, 1988 Jul, 134 ( Pt 7), 1971 - 6
Thermostable peroxidase from Bacillus stearothermophilus; Loprasert S et al.; A peroxidase from Bacillus stearothermophilus was purified to homogeneity . The enzyme (Mr 175,000) was composed of two subunits of equal size, and showed a Soret band at 406 nm . On reduction with sodium dithionite, absorption at 434 nm and 558 nm was observed . The spectrum of reduced pyridine haemochrome showed peaks at 418, 526 and 557 nm; the reduced minus oxidized spectrum of pyridine haemochrome showed peaks of 418, 524 and 556 nm with a trough at 452 nm . These results indicate that the enzyme contained protohaem IX as a prosthetic group . The optimum pH was about 6 and the apparent optimum temperature was 70 degrees C . The enzyme was relatively stable up to 70 degrees C; at 30 degrees C it was stable for a month . The enzyme had peroxidase activity toward a mixture of 2,4-dichlorophenol and 4-aminoantipyrine with a Km for H2O2 of 1.3 mM . It also acted as a catalase with a Km for H2O2 of 7.5 mM.

J Gen Microbiol, 1988 Jul, 134 ( Pt 7), 1847 - 82
A numerical classification of the genus Bacillus; Priest FG et al.; Three hundred and sixty-eight strains of aerobic, endospore-forming bacteria which included type and reference cultures of Bacillus and environmental isolates were studied . Overall similarities of these strains for 118 unit characters were determined by the SSM, SJ and DP coefficients and clustering achieved using the UPGMA algorithm . Test error was within acceptable limits . Six cluster-groups were defined at 70% SSM, which corresponded to 69% SP and 48-57% SJ . Groupings obtained with the three coefficients were generally similar but there were some changes in the definition and membership of cluster-groups and clusters, particularly with the SJ coefficient . The Bacillus strains were distributed among 31 major (4 or more strains), 18 minor (2 or 3 strains) and 30 single-member clusters at the 83% SSM level . Most of these clusters can be regarded as taxospecies . The heterogeneity of several species, including Bacillus brevis, B . circulans, B . coagulans, B . megateriun, B . sphaericus and B . stearothermophilus, has been indicated and the species status of several taxa of hitherto uncertain validity confirmed . Thus on the basis of the numerical phenetic and appropriate (published) molecular genetic data, it is proposed that the following names be recognized; Bacillus flexus (Batchelor) nom . rev., Bacillus fusiformis (Smith et al.) comb . nov., Bacillus kaustophilus (Prickett) nom . rev., Bacillus psychrosaccharolyticus (Larkin & Stokes) nom . rev . and Bacillus simplex (Gottheil) nom . rev . Other phenetically well-defined taxospecies included 'B . aneurinolyticus', 'B . apiarius', 'B . cascainensis', 'B . thiaminolyticus' and three clusters of environmental isolates related to B . firmus and previously described as 'B . firmus-B . lentus intermediates' . Future developments in the light of the numerical phenetic data are discussed.

Cytometry, 1988 Jul, 9(4), 387 - 93
Light regulation of the cell cycle in Euglena gracilis bacillaris; Yee MC et al.; We have studied the light regulation of the cell division cycle in the photosynthetic alga Euglena gracilis bacillaris . Euglena grown under phototrophic conditions are easily synchronized to a 12 h light-12 h dark regime . By inoculating stationary phase, nondividing cells into fresh media and exposing the diluted cells to either light or darkness, we have determined that initiation of DNA synthesis for the cell division cycle is light dependent . By varying the length of time in light to which synchronized cells are exposed, we have shown that commitment to the cell cycle requires exposure to more than 6 h of light . We propose that this is to allow the accumulation, through photosynthetic electron transport, of an initiating factor that will enable DNA synthesis to begin . Flow cytometry analysis also shows that once cells are committed to the cell cycle, they complete the cycle in the dark, so mitosis is a light-independent step.

J Gen Microbiol, 1988 Jul, 134 ( Pt 7), 1961 - 70
Molecular cloning and characterization of Bacillus alvei thiol-dependent cytolytic toxin expressed in Escherichia coli; Geoffroy C et al.; A chromosomal DNA fragment from Bacillus alvei, encoding a thiol-dependent haemolytic product known as alveolysin (Mr 60,000, pI 5.0) was cloned in Escherichia coli SK1592, using pBR322 as the vector plasmid . Only a single haemolysin-positive clone was identified, by testing for haemolysis on blood agar plates . The haemolytic material was associated with the host bacterial cell . It was released by ultrasonic disruption and purified 267-fold . A 64 kDa polypeptide of pI 8.2 cofractionated with haemolytic activity during gel filtration chromatography and isoelectric focusing . It behaved identically to alveolysin in its activation by thiols, inactivation by thiol group reagents, inhibition by cholesterol, and neutralization, immunoprecipitation and immunoblotting by immune sera raised against alveolysin and streptolysin O.

Clin Microbiol Rev, 1988 Jul, 1(3), 330 - 48
Leprosy; Hastings RC et al.; Leprosy affects over 10 million people in the world . The disease is a model of graded cell-mediated immunity, in this case to the causative organism, Mycobacterium leprae . The clinical manifestations are due to (i) bacterial progression, (ii) immunologic responses of the host, (iii) peripheral nerve damage due to either or both bacterial progression and immunologic responses of the host, and (iv) preventable secondary deformities following nerve damage, which account for most of the stigma of the disease . Treatment modalities are now available to control or minimize the effects of bacterial progression, harmful immunologic responses of the host, peripheral nerve damage, and secondary deformities . Unique biochemical characteristics of M . leprae reside in the cell wall and associated macromolecules . Some of these molecules are potent immunogens in humans, while others constitute the structural integrity of the bacillus . Proteins of M . leprae are currently under intensive investigation as a result of deoxyribonucleic acid cloning of M . leprae genes . Structure-function and antigenic relationships of M . leprae proteins should become available by using recombinant deoxyribonucleic acid procedures coupled with T- and B-cell cloning to advance our understanding of the immunologic reactions encountered in Hansen's disease . Until recently, the study of the immunology of leprosy has been stymied by the lack of immunologically specific M . leprae antigens . The definition of specific antigens and production of recombinant and synthetic immunologic reagents have fostered state-of-the-art research efforts into new immunodiagnostic procedures and development of a leprosy vaccine . Also discussed is progress in understanding of the mechanism(s) underlying the M . leprae-specific immunodeficiency associated with lepromatous leprosy, including the role of suppressor T cells and defective macrophage function . Metabolic studies of M . leprae suggest intact catabolic pathways and energy generation with purine bases and catalase as possible growth factors . Special attention may also need to be given to biophysical parameters for eventual in vitro cultivation . Rapid in vitro systems, using quantitation of bacillary metabolic activity, may soon replace the lengthy mouse footpad test for determining the viability and drug susceptibility of the leprosy bacillus.

J Biochem (Tokyo), 1988 Jul, 104(1), 12 - 3
Crystallization of and preliminary crystallographic data for Bacillus stearothermophilus cyclodextrin glucanotransferase; Kubota M et al.; Cyclodextrin glucanotransferase from Bacillus stearothermophilus TC-91 has been crystallized from an ammonium sulfate solution by the dialysis equilibrium method . The crystals belong to the orthorhombic system, space group P2(1)2(1)2(1), with cell dimensions of a = 125.5 A, b = 88.1 A, and c = 81.5 A . The crystals appear to be suitable for X-ray structure analysis, diffracting to at least 2.1 A and being resistant to radiation damage.

Bioorg Khim, 1988 Jul, 14(7), 916 - 20
{A new type of cleavage of the recognition site by the site-specific endonuclease Bst 4.4I from Bacillus stearothermophilus 4.4}; Kramarov VM et al.; A site-specific endonuclease Bst 4.4I was isolated from the cell extract of Bacillus stearothermophilus 4.4 and partially purified by chromatography on Ultragel AcA-44 and heparin-Sepharose . It was shown that the endonuclease cleaves lambda and M13 DNA yielding distinct fragments just as endonucleases of II and III types but, in contrast to them can produce two two-strand cuts separated with 30 to 32 nucleotides in the region of the recognition site.

Antimicrob Agents Chemother, 1988 Jul, 32(7), 1053 - 6
Selective inhibition of human cytomegalovirus replication by a novel nucleoside, oxetanocin G; Nishiyama Y et al.; A novel nucleoside with an oxetanosyl-N-glycoside has been recently isolated from a culture filtrate from Bacillus megaterium and named oxetanocin A (N . Shimada, S . Hasegawa, T . Harada, T . Tomisawa, A . Fujii, and T . Takita, J . Antibiot . 39:1623-1625, 1986) . In this study, we evaluated the antiherpesvirus activity of oxetanocin A and its derivatives and found that 9-(2-deoxy-2-hydroxymethyl-beta-D-erythro-oxetanosyl)guanine (OXT-G) was very potent and selective in inhibiting the replication of human cytomegalovirus (HCMV) in vitro . The median effective concentration for HCMV strain AD169 was 1.0 microgram/ml, and that for herpes simplex virus type 2 strain 186 was 3.5 micrograms/ml . The selectivity index, based on the ratio of the median inhibitory concentration for cell growth of human diploid fibroblasts to the median effective concentration for HCMV plaque formation, was more than 300 . The synthesis of HCMV-induced late polypeptides such as the 150,000-molecular-weight capsid and the 68,000-molecular-weight major matrix proteins was strongly suppressed when OXT-G (5 micrograms/ml) was added to the cultures at the beginning of infection . At this concentration of OXT-G, the amount of HCMV DNA detected in the drug-treated infected cells was less than 1/10 of that detected in the infected control cells . The results suggest that the mode of action of OXT-G is inhibition of viral replication by impairing the viral DNA synthesis.

Biochem J, 1988 Jul 1, 253(1), 235 - 41
Activation of insect cell adenylate cyclase by Bacillus thuringiensis delta-endotoxins and melittin . Toxicity is independent of cyclic AMP; Knowles BH et al.; Insecticidal Bacillus thuringiensis (Bt) delta-endotoxins are cytolytic to a range of insect cell lines in vitro . Addition of Bt var . aizawai or var . israelensis toxins to Mamestra brassicae (cabbage moth) cells in vitro increased intracellular cyclic AMP, which was paralleled by activation of adenylate cyclase in isolated membranes . Var . kurstaki toxin, which does not lyse M . brassicae cells, had no effect on cyclic AMP concentrations in intact cells, but was able to stimulate adenylate cyclase in membrane preparations . In contrast, the bee-venom toxin melittin, which is also cytolytic, increased intracellular cyclic AMP in whole cells, but inhibited adenylate cyclase in isolated membranes . Octopamine and forskolin also increased cyclic AMP in cells, but were not cytolytic . When added to cells at concentrations exceeding their LC90 (concentration causing 90% cell death), melittin and var . israelensis toxins caused cell lysis without a concomitant increase in intracellular cyclic AMP . Taken together, these results suggest that activation of adenylate cyclase by cytolytic toxins is a secondary effect (related perhaps to interactions of these toxins with membrane lipids) and is neither necessary nor sufficient for cytolysis.

J Exp Biol, 1988 Jul, 137, 277 - 86
Barium and calcium block Bacillus thuringiensis subspecies Kurstaki delta-endotoxin inhibition of potassium current across isolated midgut of larval Manduca sexta; Crawford DN et al.; Ba2+ and Ca2+ prevent and reverse the Btk delta-endotoxin inhibition of the short-circuit current in isolated lepidopteran midgut . These findings support the K+ pump-leak steady-state model for midgut K+ homeostasis and the K+ channel mechanism of Bt toxin action . They provide a new tool with which to study the interactions between Bt toxin and midgut cell membranes.

Prikl Biokhim Mikrobiol, 1988 Jul-Aug, 24(4), 535 - 41
{Biological properties of a Bacillus brevis 101 mutant}; Vypiiach AN et al.; Some features of the Bacillus brevis 101 mutant producing the antibiotic gramicidin S are described . The mutant is very close to the initial P+-variant of Bacillus brevis var . G-B by cultural, physiological and biochemical characteristics . The most typical features of Bacillus brevis 101 are high antibiotic activity (up to 2 g/l) and the specific phenotype of the colonies . The phenotypical features of Bacillus brevis 101 are dependent on the conditions of its cultivation . On minimal media rich in organics a change of the culture correlated with a lower antibiotic activity.

Nature, 1988 Jun 23, 333(6175), 784 - 6
Contribution of hydrophobic interactions to protein stability; Kellis JT Jr et al.; A major factor in the folding of proteins is the burying of hydrophobic side chains . A specific example is the packing of alpha-helices on beta-sheets by interdigitation of nonpolar side chains . The contributions of these interactions to the energetics of protein stability may be measured by simple protein engineering experiments . We have used site-directed mutagenesis to truncate hydrophobic side chains at an alpha-helix/beta-sheet interface in the small ribonuclease from Bacillus amyloliquefaciens (barnase) . The decreases in stability of the mutant proteins were measured by their susceptibility to urea denaturation . Creation of a cavity the size of a -CH2-group destabilizes the enzyme by 1.1 kcal mol-1, and a cavity the size of three such groups by 4.0 kcal mol-1.

Biochem J, 1988 Jun 15, 252(3), 661 - 6
A new alcohol dehydrogenase, reactive towards methanol, from Bacillus stearothermophilus; Sheehan MC et al.; An NAD+-dependent alcohol dehydrogenase (ADH) was purified to homogeneity from an aerobic strain of Bacillus stearothermophilus, DSM 2334 (ADH 2334), and compared with the ADH from B . stearothermophilus NCA 1503 (ADH 1503) . When an antibody raised against ADH 2334 was used, no cross-reactivity with ADH 1503 was observed on Western blots; by means of an enzyme-linked-immunoabsorbent-assay ('e.l.i.s.a.') procedure, it was found that ADH 1503 had less than 6% of the antigenic activity of ADH 2334 . Amino acid analyses detected very small differences in composition, equivalent to about 40 sequence changes, between the two enzymes . The new enzyme has the same six-amino-acid N-terminal sequence as ADH 1503 . ADH 2334, but not ADH 1503, is reactive towards methanol; both enzymes can oxidize ethanol, propan-1-ol, butan-1-ol and butan-2-ol . The new enzyme has a distinctive pH optimum at pH 5.5-6 and has significantly lower KEthanolm and kEthanolcat . values than those of ADH 1503 . From steady-state kinetic parameters of the reaction with ethanol, propan-1-ol and butan-1-ol, it was shown that ADH 2334 has an ordered mechanism in both directions, with NAD+ being the compulsory first substrate in alcohol oxidation and NADH release being the rate-limiting step . ADH 1503 has an ordered addition of NAD+ and alcohol, but NADH release is not rate-limiting.

J Biol Chem, 1988 Jun 15, 263(17), 8215 - 9
Amiloride, a specific inhibitor for the Na+-driven flagellar motors of alkalophilic Bacillus; Sugiyama S et al.; Na+-driven flagellar motors of alkalophilic Bacillus were found to be inhibited by amiloride, a potent inhibitor for many Na+-coupled systems . A concentration of 0.5 mM of amiloride completely inhibited motility but showed almost no effect on the membrane potential, the intracellular pH homeostasis, and the ATP content of the cells . Furthermore, the activity of a Na+-coupled amino acid transport system was reduced only by half by this concentration of amiloride . Thus, the inhibition of motility of alkalophilic Bacillus by amiloride was rather specific . The inhibition of motility produced by amiloride was restored by increasing Na+ concentrations in the medium . Kinetic analysis of the data revealed that the inhibition was competitive with respect to the concentration of Na+ in the medium . Therefore, it is quite logical to assume that amiloride inhibits the rotation of the Na+-driven flagellar motors of alkalophilic Bacillus by competing with Na+ at the force-generating site of the motor . Some amiloride analogs known to selectively inhibit Na+ channels were potent inhibitors for the flagellar motors, suggesting that the Na+-interacting site of the motors has some similarity to that of the Na+ channels.

Gene, 1988 Jun 15, 66(1), 107 - 20
Nucleotide sequence of the gene coding for a 130-kDa mosquitocidal protein of Bacillus thuringiensis israelensis; Yamamoto T et al.; The nucleotide sequence of pVB131 containing the gene coding for a 130-kDa Bacillus thuringiensis israelensis (B.t.isr) mosquitocidal protein was determined . The pVB131 plasmid was constructed by Sekar and Carlton {Gene 33 (1985) 151-158} . Our sequencing revealed only one open reading frame large enough to code for a protein of 130 kDa . The translation start site was determined by sequencing the protein isolated from B.t.isr . The amino acid sequence of the protein was deduced from the nucleotide sequence, and its Mr was determined as 128,505 . Immunological and biochemical analyses of B.t.isr mosquitocidal proteins indicated that the 130-kDa protein coded by pVB131 was indeed expressed in B.t.isr . Comparing the peptide sequence of the 130-kDa B.t.isr toxin with the sequences of other B.t . toxins having activities specific to lepidopteran species showed that several domains were highly homologous . This suggests that they are evolutionarily related to each other, and in the evolutionary process the sequences in the homologous domains that are important to the insecticidal activity have been conserved.

FEBS Lett, 1988 Jun 6, 233(1), 119 - 23
Three-dimensional location of ribosomal protein BL2 from Bacillus stearothermophilus, a key component of the peptidyl transferase center; Hackl W et al.; Protein BL2 from Bacillus stearothermophilus has been localized by immunoelectron microscopy on the interface side of the 50 S subunit, beneath the angle formed between the central protuberance and the L1 protuberance . The immuno-electron microscopic data suggest that the interface region of the 50 S particle is not as flat as most of the proposed three-dimensional models suggest, but instead there is a significant concavity . Since several studies demonstrated that BL2 is implicated in peptidyl transferase activity or at least located close to the peptidyl transferase center, the location of protein BL2 also provides information as to the location of this important functional domain.

J Biol Chem, 1988 Jun 5, 263(16), 7895 - 906
The three-dimensional structure of Bacillus amyloliquefaciens subtilisin at 1.8 A and an analysis of the structural consequences of peroxide inactivation; Bott R et al.; The three-dimensional structure of the subtilisin from Bacillus amyloliquefaciens (BAS) has been refined to 1.8 A using the amino acid sequence deduced from the DNA coding sequence . The structure is essentially the same as the previously reported structures of subtilisin BPN' (Wright, C.S., Alden, R.A., and Kraut, J . (1969) Nature 221, 235-242) and Novo (Drenth, J., Hol, W . G . J., Jansonius, J . N., and Koekoek, R . (1972) Eur . J . Biochem . 26, 177-181) determined in different crystal forms, at 2.5 and 2.8 A resolution, respectively . The largest differences in the three crystallographic models are seen in regions where the amino acid sequence used in the fit to the electron density maps of BPN' and Novo differs from the gene sequence of BAS (Wells, J . A., Ferrari, E., Henner, D . J., Estell, D . A., and Chen, E . Y . (1983) Nucleic Acids Res . 11, 7911-7925) . The refined BAS model shows new features of cation binding, hydrogen bonding, and internal solvent structure . The refined BAS model has served as a basis for the analysis of stereochemical factors involved in the peroxide inactivation of the enzyme . Methionine 222, which is adjacent to the catalytic Ser221, is quantitatively oxidized to the sulfoxide by hydrogen peroxide as had been previously shown for the related Bacillus licheniformis enzyme (Stauffer, C . E., and Etson, D . (1969) J . Biol . Chem . 244, 5333-5338) . In addition to this site of modification, we observe partial to full oxidation of two of the four remaining methionines . The oxidation of the methionines does not correlate well with their solvent accessibility calculated from the x-ray structure coordinates; in addition, only one of the two possible stereoisomers of methionine sulfoxide is formed . We also detect hydrogen peroxide-induced modification of the hydroxyl groups of two tyrosines . Modeling suggests that most of the observed effect of oxidation on the enzyme's catalytic efficiency can be attributed to unfavorable interactions at the oxyanion binding site between the sulfoxide group at 222 and the carbonyl oxygen of the scissile peptide bond of the bound substrate.

Eur J Biochem, 1988 Jun 1, 174(2), 255 - 60
Pyruvic-acid-containing polysaccharide in the cell wall of Bacillus polymyxa AHU 1385; Kojima N et al.; Three acidic polymer fractions with molecular masses of about 16 kDa, 35 kDa and 70 kDa were isolated from lysozyme digests of N-acetylated cell walls of Bacillus polymyxa AHU 1385 by ion-exchange chromatography and gel chromatography . These fractions, containing mannosamine, glucosamine and pyruvic acid in a molar ratio of about 1:1:1 together with glycopeptide components, were characterized as polysaccharide-linked glycopeptides with one, two and more polysaccharide chains . On the other hand, treatment of the cell walls with glycine/HC1 buffer, pH 2.5, at 100 degrees C for 10 min followed by separation of water-soluble products on ion-exchange chromatography gave three polysaccharide fractions, PS-I-III, which contained different amounts of pyruvic acid (0,0.6 and 0.9 residue/mannosamine residue) along with equimolar amounts of mannosamine and glucosamine . Pyruvate-free polysaccharides similar to PS-I were also obtained from PS-II, PS-III and polysaccharide-linked glycopeptides by treatment with 10 mM HC1 at 100 degrees C for 1 h . Results of analyses of these polysaccharide preparations by 1H-NMR and 13C-NMR measurement and methylation, together with data from characterization of fragments obtained by hydrogen fluoride hydrolysis, lead to the most likely structure, ----3){4,6-O-(1-carboxyethylidene)}ManNAc(beta 1----4)GlcNac(beta 1----, for the acidic polysaccharide of this strain.

Urology, 1988 Jun, 31(6), 459 - 68
Does bacillus Calmette-Guérin immunotherapy accelerate growth and cause metastatic spread of second primary malignancy?
Khanna OP, Chou RH, Son DL, Mazer H, Read J, Nugent, Cottone R, Heeg M, Rezvan M, Viek N, et al.
In our study, 29 of 150 patients with bladder cancer also had other associated primary malignancies, 10 of which were manifested after intravesical treatment with bacillus Calmette-Guerin (BCG) . Second primary malignancies developed in 5 of these patients within three months of the start of BCG therapy . All 5 showed acceleration of the second primary tumor, and distant metastatic lesions developed in 4 . In the other 5 patients nonbladder primary malignancies developed eight months or more after intravesical BCG therapy started, but did not show acceleration or spread . Twenty patients with other primary malignancies that had developed months to years before intravesical therapy did not show acceleration or spread of those tumors . We have seen enough cases of patients who received intravesical BCG at the time of growth and spread of second primary malignancies to warrant concern . Animal and human studies of BCG use for treatment of malignancy indicate that the temporal relationship between the starting point of tumor development and the starting point of BCG treatment is crucial in determining whether BCG will eradicate or exacerbate the tumor . We have therefore instituted a change in our treatment until the question of whether or not BCG causes the appearance and spread of these second malignancies is answered.

J Clin Microbiol, 1988 Jun, 26(6), 1124 - 9
Biophysical optima for metabolism of Mycobacterium leprae; Franzblau SG et al.; The metabolic response of freshly harvested, nude-mouse-derived Mycobacterium leprae to biophysical parameters was studied to facilitate an understanding of axenic culture requirements . Quantitation of intracellular ATP and the rate of {U-14C}palmitic acid incorporation into phenolic glycolipid I (PGL-I) were used as metabolic indicators after axenic incubation in modified Dubos medium under various biophysical conditions . PGL-I synthesis was optimal at 33 degrees C, whereas ATP was optimally maintained at less than or equal to 33 degrees C . Both metabolic indices showed sharp reductions at 37 degrees C . After 5 days of incubation, PGL-I synthesis and ATP maintenance showed pH optima of 5.1 to 5.6, with the higher value appearing optimal for ATP maintenance after extended incubation . Metabolic activity was negatively affected by strong reducing agents, and ATP maintenance was optimal when the gaseous environment was maintained at 2.5 to 10% oxygen . The results may partially explain the failure to cultivate the leprosy bacillus in vitro.

Eur J Biochem, 1988 Jun 1, 174(2), 431 - 5
Immunoelectron microscopic localisation of ribosomal proteins from Bacillus stearothermophilus that are homologous to Escherichia coli L1, L6, L23 and L29; Hackl W et al.; The locations of proteins BL1, BL6, BL23 and BL29 from Bacillus stearothermophilus have been determined on the ribosomal surface by immunoelectron microscopy . All four proteins were localized in the same region of the 50S subunit as their homologous counterparts from Escherichia coli, indicating that the ribosomal architecture is the same in both species . This finding is of great importance as it allows structural data obtained on ribosomes from either organism to be incorporated into a unique ribosome model.

J Vasc Surg, 1988 Jun, 7(6), 808 - 10
Mycotic abdominal aortic aneurysm induced by immunotherapy with bacille Calmette-Guérin vaccine for malignancy; Woods JM 4th et al.; Successful surgical treatment of a mycotic abdominal aortic aneurysm infected with Mycobacterium bovis is described . The infecting organism can be traced to an intraneoplastic injection of bacille Calmette-Guerin (BCG) vaccine into a cutaneous malignant melanoma nodule 14 months before aneurysm detection (17 months before operation) . Treatment consisted of aneurysm excision, in situ prosthetic graft placement, and antituberculous medications . This patient represents the first reported case of BCG-induced mycotic aortic aneurysm.

Pediatr Infect Dis J, 1988 Jun, 7(6), 375 - 9
Tuberculosis in children and adolescents in the 1980s; Nemir RL et al.; Tuberculosis (TBC) continues to be a major health problem . Between January 1, 1980, and April 30, 1986, 211 children and adolescents presented with a positive tuberculin reaction or symptoms suggestive of tuberculosis . Active disease occurred in 35 (17%); 29 of these had primary infection, whereas 4 adolescents presented with cavitary pulmonary disease and 1 infant each had Pott's disease and cervical adenitis . The proportion of patients with active disease was greater in infants and toddlers; 2 of whom also had meningitis . Two children with active disease were infected with human immunodeficiency virus, 1 of whom died with cavitary tuberculosis . Only 43% of 211 patients were born in the United States . The ethnic distribution was Hispanic 45%, Oriental 30%, Black 18% and other 7% . Bacillus Calmette-Guerin vaccination was documented in 53 (25%) patients; 5 (9%) of these developed active disease . Despite vaccination and the availability of effective drugs, tuberculosis persists and appears to be increasing . Meeting the challenge of tuberculosis in the future will require more rapid diagnostic methods and recognition of the burden of infection in human immunodeficiency disease-infected children, together with revitalization of screening and follow-up programs, especially for toddlers and adolescents.

No Shinkei Geka, 1988 Jun, 16(7), 907 - 9
{A case of hemorrhage into a brain abscess}; Fujii M et al.; A rare case of hemorrhage into a brain abscess in a 23-year-old man is reported . The patient complained of headache and low-grade fever on February 26, 1986 . Two days later, he developed right hemiparesis and right hemisensory disturbance with mild consciousness disturbance and was admitted to a local hospital . Seven days after the onset, he suddenly became semicomatose, developed anisocoria and was consequently transferred to the University Hospital . On admission, his temperature was 37.5 degrees C and neurological examination revealed semicoma, anisocoria and right hemiparesis without nuchal rigidity . Enhanced CT scan showed a high density area within an irregular ring enhancement at the left basal ganglia . At that time, malignant glioma was diagnosed and an emergency operation was performed by left frontotemporal craniectomy . During the operation blood clot was found in the posterior part of the basal ganglia . After operation, a histological examination was made and a brain abscess was diagnosed . Gram staining revealed gram-positive bacillus . By aspiration of the abscess and chemotherapy, recovery was gradually made . He was discharged with motor dysphasia and mild right hemiparesis three months later . Differentiation between abscess and malignant glioma and the cause of the hemorrhage are discussed.

J Gen Microbiol, 1988 Jun, 134 ( Pt 6), 1577 - 85
Five unique temperate phages from a polylysogenic strain of Bacillus thuringiensis subsp . aizawai; Reynolds RB et al.; Five temperate phages were isolated from strain 4042B of Bacillus thuringiensis subsp . aizawai . The phages, which were heteroimmune, could also be distinguished by their host ranges, plaque and particle morphologies, serological specificities, and locations of restriction endonuclease cleavage sites on their chromosomes . Besides maintaining a stable lysogenic relationship with the 4042B host strain, each phage formed a stable lysogen with Bacillus cereus.

J Am Mosq Control Assoc, 1988 Jun, 4(2), 172 - 4
Efficacy of Bacillus sphaericus 2362 formulations against floodwater mosquitoes; Mulla MS et al.; Four new formulations of Bacillus sphaericus 2362 yielded excellent control of floodwater mosquitoes Psorophora columbiae and Aedes nigromaculis in irrigated fields in Kings and Riverside counties of California . A primary powder formulation (ABG-6184) was the most active, producing excellent control of Ps . columbiae and Ae . nigromaculis at the rates of 0.05 to 0.5 lb/acre (0.055-0.56 kg/ha) . A liquid formulation (BSP-2) was slightly less active, but was effective against the same species in the range of 1.0-1.5 lb/acre (1.12-1.68 kg/ha) . Activity of the two corn cob granular formulations was largely dependent on potency (spores/gram) . The high spore count granules (1.5 x 10(9) spores/gram) yielded 91 and 98% reduction of Ae . nigromaculis at the rates of 2.5 and 5.0 lb/acre (2.8 and 5.6 kg/ha), respectively . Against the same population, the lower spore count formulation (7.6 x 10(8) spores/gram) produced complete control at the rate of 10 lb/acre (11.2 kg/ha), but poor results were obtained at the rate of 5 lb/acre (5.6 kg/ha).

J Am Mosq Control Assoc, 1988 Jun, 4(2), 132 - 7
Method for determining settling rates of Bacillus thuringiensis serotype H-14 formulations; Mullen GR et al.; A water-column apparatus is described in which settling rates of Bacillus thuringiensis serotype H-14 {B.t . (H-14)} formulations can be indirectly quantified using mortality of mosquito larvae at restricted depths as an index of B.t . (H-14) activity . To illustrate the type of data provided by this method, commercial B.t . (H-14) products (Bactimos, Teknar, Vectobac) and experimental formulations were compared at the manufacturers' recommended rates for mosquito control . All evaluations utilized laboratory-reared, 4th-instar Aedes aegypti larvae . The procedure can be used to provide an index of suspension properties of different B.t . (H-14) formulations and to measure dispersion rates of granular formulations resting at the bottom of a water column . Standardization of the method provides a convenient and practical means of generating comparative data on the effectiveness of B.t . (H-14) and other mosquito larvicides against specific target species.

Bioorg Khim, 1988 Jun, 14(6), 783 - 9
{Primary structure of intracellular serine proteinase from Bacillus amyloliquefaciens . I . Isolation of the enzyme and amino acid sequence of peptides of tryptic hydrolysate}; Surova IA et al.; Method of isolation of intracellular serine protease was modified . Gramicidin S-sepharose CL-4B with a higher content of the ligand, synthesized through a modified procedure, was used as an affinity sorbent which simplified the purification and led to the pure enzyme with high specific activity and 90% yield . Trypsin hydrolyzate of the protease was separated by ion-exchange chromatography on a sulphocationite resin followed by paper chromatography and paper electrophoresis to yield twenty-five individual peptides . Their complete or partial sequences, corresponding in total to 146 amino acid residues, were determined by the manual Edman procedure.

Tubercle, 1988 Jun, 69(2), 81 - 94
Immune status in tuberculosis and response to treatment; Onwubalili JK et al.; Thirty hospitalized patients with newly diagnosed tuberculosis were studied prospectively with a range of in vitro and in vivo tests of immune function . Responses were compared with those of healthy controls matched for age, sex, ethnic group and diet . A series of metabolic and immunologic abnormalities was found, including evidence of undernutrition, anaemia, neutrophil leucocytosis, monocytosis, lymphopenia, hyperglobulinaemia and raised erythrocyte sedimentation rate . Some patients had accelerated, others diminished, cutaneous tuberculin hypersensitivity, and some had diminished mononuclear cell proliferative and lymphokine responses to tuberculin (purified protein derivative, PPD) . The patients were not uniform in their responsiveness, but could be arranged within a spectrum which showed a relationship to crude bacillary excretion and response to treatment . 27% of patients were characterized by hypersensitivity, with normal in vitro cellular responses and skin tests to PPD, scanty bacillary excretion and rapid bacteriologic sputum conversion to negative cultures with treatment . In contrast, 30% of patients were relatively anergic with negative skin tests, reduced or absent in vitro cellular reactivity to PPD, moderate or heavy bacillary excretion and later (greater than 4 weeks) bacteriologic sputum conversion . The remainder of the patients fell between these two groups . There were no correlations between cellular immunity on the one hand, and radiological extent of disease, levels of serum immunoglobulins, peripheral white cell counts or ESR on the other . In those patients followed throughout treatment, all the abnormalities with the exceptions of arm muscle circumference and serum albumin, reverted to the normal ranges established in the control group.

Arch Biol Med Exp (Santiago), 1988 Jun, 21(1), 101 - 7
The relationship between the structures of four beta-lactamases obtained from Bacillus cereus; Cid H et al.; Bacillus cereus has proved to be one of the most interesting microorganisms in the study of beta-lactamases . It secrets these enzymes very efficiently and, frequently, in multiple forms . Three different forms are produced by strain 569/H; mutant 5/B of the same microorganism is constitutive for the secretion of beta-lactamases I and II . The present study, based on secondary structure prediction by two independent methods, states the relationship among the structures of beta-lactamases I, II and III produced by B . cereus 569/H and beta-lactamase I from the strain 5/B of this microorganism . A strong similarity is also established for the enzyme type III of B . cereus and the enzyme type I produced by B . licheniformis which could have an evolutionary explanation . A structural analysis of the leader peptide regions of these enzymes by the method of Mohana and Argos is also reported.

Mol Cell Probes, 1988 Jun, 2(2), 111 - 24
DNA probes for mycobacteria . I . Isolation of DNA probes for the identification of Mycobacterium tuberculosis complex and for mycobacteria other than tuberculosis (MOTT); Picken RN et al.; Traditional methods used in identifying mycobacteria such as acid-fast bacillus stains and culture are often time-consuming, insensitive and non-specific . The isolation of DNA probes, coupled to a non-radioactive, e.g . biotin-based detection system, have the potential to foster the development of clinical assays for Mycobacterium tuberculosis and mycobacteria other than tuberculosis (MOTT) that are rapid, sensitive and specific . To this end, we have isolated two different probes: one which is specific for the Mtb complex and one which recognizes all other potentially pathogenic mycobacteria . The use of these probes in combination should allow the detection and differentiation of M . tuberculosis from MOTT . To isolate the first probe, we prepared a library of M . tuberculosis DNA fragments in a lambda EMBL phage vector . Recombinant phage were screened by plaque-lift hybridization procedures using nick-translated mycobacterial genomic DNA to identify sequences specific to the Mtb complex . Inserts from candidate recombinant phage were purified, nick-translated and hybridized against a wide variety of filter-bound mycobacterial and non-mycobacterial DNAs . Two clones were identified which hybridized to the closely related M . tuberculosis, M . bovis and M . microti but not to other species of mycobacteria . The second probe was isolated by preparing a library of M . malmoense DNA fragments in lambda EMBL and screening by plaque-lift hybridization . One clone was identified which, in addition to recognizing members of the Mtb complex, also hybridized to M . intracellulare, M . malmoense, M . scrofulaceum, M . simiae, M . xenopi, M . avium, M . szulgai, M . kansasii and M . haemophilum . None of the three clones hybridized to DNA from non-mycobacterial species.

Biochem J, 1988 Jun 1, 252(2), 595 - 600
Malate dehydrogenases in phototrophic purple bacteria . Thermal stability, amino acid composition and immunological properties; Tayeh MA et al.; Purified malate dehydrogenases from four species of non-sulphur purple phototrophic bacteria were examined for their heat-stability, amino acid composition and antigenic relationships . Malate dehydrogenase from Rhodospirillum rubrum, Rhodobacter capsulatus and Rhodomicrobium vannielii (which are all tetrameric proteins) had an unusually high glycine content, but the enzyme from Rhodocyclus purpureus (which is a dimer) did not . R . rubrum malate dehydrogenase was extremely heat-stable relative to the other enzymes, withstanding 65 degrees C for over 1 h with no loss of activity . By contrast, malate dehydrogenase from R . vannielii lost activity above 35 degrees C, and that from R . capsulatus above 40 degrees C . Amino acid compositional relatedness and immunological studies indicated that tetrameric phototrophic-bacterial malate dehydrogenases were highly related to one another, but only distantly related to the tetrameric enzyme from Bacillus . This suggests that, despite differences in their thermal properties, the tetrameric malate dehydrogenases of non-sulphur purple bacteria constitute a distinct biochemical class of this catalyst.

J Inorg Biochem, 1988 Jun, 33(2), 77 - 89
Synthesis of some 5-azo(4'-substituted benzene-sulphamoyl)-8-hydroxyquinolines with antidotal and antibacterial activities; Awad IM et al.; 5-Azo(4'-substituted benzenesulphamoyl)-8-hydroxyquinolines(III) have been prepared by coupling of the appropriate p-substituted benzenesulphamoyldiazonium acetates with 8-hydroxyquinoline . The corresponding copper chelates(IV) and iron chelates(V) were also prepared in a 1:2 metal to ligand ratio . Structures of III, IV and V were confirmed by some representative UV, IR, and NMR spectrometry in addition to microanalysis . Antidotal activity of four ligands (IIIa, IIId, IIIf, and IIIi) has been evaluated in mice against the toxicity of lead acetate and copper sulphate . Study revealed that compound IIIf elicited significant antidotal activity against lead and copper poisoning, while IIIi was potent only against lead poisoning . Antibacterial activity of compounds III, IV, and V was also determined in comparison to sulphanilamide against Staph . aureus, Bacill . cereus, and Esch . coli . The test compounds showed variable bacteriostatic activities, and some of them (IIIc, IIId, IIIf, Ve, IIIg, and Vi) are more effective than the reference drug, especially against Bacill . cereus.

J Bacteriol, 1988 Jun, 170(6), 2873 - 8
Cloning, nucleotide sequence, and expression of the Bacillus cereus 5/B/6 beta-lactamase II structural gene; Lim HM et al.; Two forms of heat-stable, zinc-containing beta-lactamase II have been described for strains of Bacillus cereus and have been shown to differ in substrate specificity (R . B . Davies, E . P . Abraham, J . Fleming, and M . R . Pollock, Biochem . J . 145: 409-411, 1975) . We report here the nucleotide sequence, inferred amino acid sequence, and expression of beta-lactamase II from B . cereus 5/B/6 and compare our results with those for its homolog characterized in B . cereus 569/H (M . Hussain, C . Anthony, M . J . Madonna, and J . O . Lampen, J . Bacteriol . 164: 223-229, 1985) to document amino acid differences contributing to the specific properties of these enzymes.

J Immunol, 1988 Jun 1, 140(11), 3956 - 61
Mycobacterium lepraemurium activates macrophages but fails to trigger release of superoxide anion; Mor N et al.; Mycobacterium lepraemurium failed to stimulate a normal respiratory burst when presented to mouse peritoneal or bone marrow macrophages . By comparison, Mycobacterium bovis (strain Bacillus Calmette-Guerin) or Saccharomyces cerevisiae, as expected, stimulated macrophages to release a large amount of superoxide anion (O2-) . M . lepraemurium did not interfere with the response to yeast when both microbes were added together to macrophages . The low release of O2- induced by M . lepraemurium was not due to failure of M . lepraemurium to activate or prime macrophages, because exposure of macrophages to M . lepraemurium caused the expected enhancement of O2- release when the macrophages were stimulated by PMA . Similarly, macrophages taken from mice infected with M . lepraemurium were activated, as indicated by high PMA-stimulated O2- release . Macrophages primed in vitro by exposure to Escherichia coli LPS for 24 h did show a moderate O2- response when stimulated by M . lepraemurium, but macrophages primed by exposure to IFN-gamma muramyl dipeptide, or M . lepraemurium showed a weak response when subsequently challenged with M . lepraemurium . The priming effect of M . lepraemurium or LPS decreased substantially after macrophages were cultured in fresh medium for 24 h . Heat killing or opsonization of M . lepraemurium caused the M . lepraemurium to stimulate a high amount of O2- release from LPS-primed macrophages, but heat killing or opsonization of M . lepraemurium had no effect on release of O2- from unprimed macrophages . The results suggest that M . lepraemurium is taken into macrophages by a mechanism that bypasses the FcR and other receptors that are capable of triggering the production of O2-.

Scand J Gastroenterol, 1988 Jun, 23(5), 611 - 9
Whipple's disease . Demonstration of a persisting monocyte and macrophage dysfunction; Bjerknes R et al.; A patient with Whipple's disease has been followed up for 4 years . Primary involvement was limited to the small intestines, and accumulation of periodic acid-Schiff-positive material, containing typical more or less intact bacillary bodies, was demonstrated within macrophages of affected tissue . After initial oxytetracycline treatment and clinical remission, the patient relapsed, with multiorgan affections . The antibiotic regimen was changed to chloramphenicol, followed by continuous trimethoprim-sulfamethoxazole . Flow cytometric studies showed persisting impairment of monocyte and macrophage intracellular degradation of bacteria during all the 4 years tested . After relapse, reduced activity of several brush border enzymes was demonstrated in distal duodenal biopsy specimens . After 17 months of continuous trimethoprim-sulfamethoxazole therapy complete clinical remission, regression of histopathologic abnormalities, and restoration of duodenal enzyme activities had occurred . The results demonstrate a persisting dysfunction of mononuclear phagocytes from a patient with Whipple's disease, suggesting a primary abnormality of cell-mediated immunity which may promote the susceptibility to the causative bacillus.

Biochem Biophys Res Commun, 1988 May 31, 153(1), 294 - 300
The mosquito larvicidal activity of 130 kDa delta-endotoxin of Bacillus thuringiensis var . israelensis resides in the 72 kDa amino-terminal fragment; Pao-intara M et al.; Bacillus thuringiensis var . israelensis produces 130 kDa delta-endotoxin which is highly toxic to mosquito-larvae . The mosquito-larvicidal activity was delineated by sequential deletions from ends of the 1136 amino acids delta-endotoxin . A maximum of 459 amino acids could be removed from the carboxy-terminal of the toxin without a significant loss of the larvicidal activity . However, no more than 38 amino acids could be deleted from the amino-terminal without losing the toxicity . The truncated peptide of 72 kDa exhibited similar toxicity to the 130 kDa toxin and was between 39th and 677th amino acids.

Gene, 1988 May 30, 65(2), 293 - 304
Cloning and sequencing of the gene encoding the phosphatidylcholine-preferring phospholipase C of Bacillus cereus; Johansen T et al.; A synthetic oligodeoxynucleotide probe was used to clone the gene encoding the phosphatidylcholine-preferring phospholipase C of Bacillus cereus . The sequence of a 2050-bp restriction fragment containing the gene was determined . Analysis of the gene-derived amino acid (aa) sequence showed that this exoenzyme is probably synthesized as a 283-aa precursor with a 24-aa signal peptide and a 14-aa propeptide . The mature, secreted enzyme comprises 245 aa residues . Sonicates of Escherichia coli HB101 carrying the gene on a multicopy plasmid showed phospholipase C activity . This activity was inhibited by Tris, a known inhibitor of the B . cereus enzyme and also by antiserum raised against pure B . cereus phospholipase C . We conclude therefore that the gene is expressed in E . coli . The cloning and sequencing described here complete the first step toward using in vitro mutagenesis for investigations of the structure-function relationships of B . cereus phospholipase C.

Lancet, 1988 May 21, 1(8595), 1132 - 6
Granulomatous hepatitis associated with cat scratch disease; Lenoir AA et al.; In three patients with cat scratch disease the liver was affected . All three had high fever (39 degrees C) for more than 3 weeks . Two of them had no peripheral adenopathy . Computed tomography of the abdomen revealed focal hepatic defects in two patients and periportal and periaortic adenopathy in the third . At laparotomy, there were nodules on the liver surfaces of all patients and histological examination revealed necrotising granulomata . The Warthin-Starry silver stain showed organisms consistent in appearance with the cat scratch bacillus in the liver and a periaortic lymph node of one patient, in the liver of the second patient, and in the axillary lymph node of the third . In all three patients the clinical findings and radiological abnormalities improved without specific therapy . A review of the surgical pathology files of Washington University revealed only two other cases of granulomatous hepatitis in children over a 6-year period . These findings indicate that cat scratch disease should now be included in the differential diagnosis of granulomatous hepatitis, at least in children . The absence of peripheral adenopathy in two of the three patients with granulomatous hepatitis suggests that the clinical spectrum of cat scratch disease may be broader than previously appreciated.

Biochem J, 1988 May 15, 252(1), 79 - 86
Amino acid sequence analysis of the lipoyl and peripheral subunit-binding domains in the lipoate acetyltransferase component of the pyruvate dehydrogenase complex from Bacillus stearothermophilus; Packman LC et al.; The pyruvate dehydrogenase multienzyme complex from Bacillus stearothermophilus comprises a structural core, composed of 60 dihydrolipoamide acetyltransferase (E2p) subunits, which binds multiple copies of pyruvate decarboxylase (E1p) and dihydrolipoamide dehydrogenase (E3) subunits . After limited proteolysis with chymotrypsin, the N-terminal lipoyl domain of E2p was excised, purified and sequenced . The residual complex, which remained assembled, was then digested with trypsin under mild conditions . This treatment promoted complete disassembly of the complex and the various components were separated by gel filtration and h.p.l.c . A folded fragment of E2p containing about 50 amino acid residues was identified as being responsible for binding the E3 subunits, although, unlike the corresponding region of the E2p or E2o chains of the pyruvate dehydrogenase or 2-oxoglutarate dehydrogenase complexes from Escherichia coli, the fragment also bound E1p molecules . Further peptide purification and sequence analysis allowed the determination of the first 211 amino acid residues of the B . stearothermophilus E2p chain, thus providing the complete primary structure of the lipoyl domain, the E1p/E3-binding domain and the regions of polypeptide chain, probably highly flexible in nature, that link the domains to each other and to the inner-core (E2p-binding) domain . Several of the proteolytically sensitive sites were also identified . The sequence of the B . stearothermophilus E2p chain shows close homology with the sequences of the E2p and E2o chains from E . coli, although significant differences in structure are apparent . Detailed evidence for the sequence of the peptides obtained by limited proteolysis and further chemical and enzymic cleavages have been deposited as Supplementary Publication SUP 50142 (11 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 6BQ, U.K., from whom copies may be obtained as indicated in Biochem . J . (1988) 249, 5.

Arch Biochem Biophys, 1988 May 15, 263(1), 121 - 9
Kinetic evidence for a reversible isomerization of pig muscle glyceraldehyde-3-phosphate dehydrogenase in its crystallization medium; Vas M et al.; Ammonium sulfate, a typical component of crystallization media of proteins, stabilizes an inactive conformation of pig muscle glyceraldehyde-3-phosphate dehydrogenase . In fact, in the presence of ammonium sulfate the reconstitution of the catalytically active holoenzyme from the apoenzyme and NAD is not instantaneous, as in the case of enzymes from Bacillus stearothermophilus and the Mediterranean lobster Palinurus vulgaris . With pig muscle enzyme, at pH 6.0, the time course of formation of the characteristic Racker band can be monitored by a rapid mixing stopped flow technique . Activation follows a single exponential curve with a rate constant independent of the concentration of both NAD and protein and, therefore, appears to be limited by a slow protein isomerization (k = 7 +/- 2 s-1) . Accordingly, when the apoenzyme is simultaneously exposed to NAD and either glyceraldehyde 3-phosphate or 1,3-bisphosphoglycerate, the ensuing reactions (the redox and the acylation steps, respectively) are kinetically limited by the same protein isomerization . At pH 7.0 and 8.0, however, two among the four active sites react with NAD at an unmeasurably high rate, while the other two sites behave as they do at pH 6.0 . When the pig muscle apoenzyme is preincubated and allowed to react with either glyceraldehyde 3-phosphate or 1,3-bisphosphoglycerate before the rapid mixing with NAD, both the redox reaction and the NAD-dependent activation of apo-acyl-enzyme toward arsenolysis become unmeasurably fast . Similarly, when the sulfate in the medium is replaced by ions such as phosphate and citrate, the reconstitution of the active holoenzyme is practically instantaneous . Thus, the slow protein isomerization observed in the presence of sulfate and abolished by competing substrates and anions is diagnostic of a structural state of the pig muscle apoenzyme, which is induced by sulfate ions bound within the enzyme active site.

Biochim Biophys Acta, 1988 May 11, 933(3), 470 - 7
Purification and characterization of two soluble cytochromes from the alkalophile Bacillus firmus RAB; Davidson MW et al.; A soluble cytochrome c and soluble cytochrome b were purified from the alkalophilic Bacillus firmus RAB . The cytochrome c, with an alpha band at 552 nm, had an apparent molecular weight of 16,500 and was acidic, with a pI of 3.4 . At both pH 7.0 and 8.3, the midpoint potential of c-552 was +66 mV . Above pH 8.3, the cytochrome exhibited a pH-dependent decrease in midpoint potential . This property, among others, distinguished the cytochrome c-552 from other membrane-associated c-type cytochromes . The soluble cytochrome b, with an alpha band maximum at 558 nm, had a molecular weight of approx . 15,500 and was also an acidic protein, with a pI of 3.07 . It exhibited a pH-independent midpoint potential of +28 mV.

Biochemistry, 1988 May 3, 27(9), 3372 - 81
Cross-linked amino acids in the protein pair S13-S19 and sequence analysis of protein S13 of Bacillus stearothermophilus ribosomes; Brockmoller J et al.; The 30S ribosomal subunits from Bacillus stearothermophilus were cross-linked under native conditions with the bifunctional reagent diepoxybutane . The dominant protein-protein cross-link in the 30S ribosomal subunit between proteins S13 and S19 {Brockmoller, J., & Kamp, R.M . (1986) Biol . Chem . Hoppe-Seyler 367, 925-935} was isolated on a preparative scale . The presence of a single cross-link site between cysteine-83 of protein S13 and histidine-68 of protein S19 was established by microsequence analysis of isolated cross-linked peptides . This cross-link site was further confirmed by different analytical methods including fast atom bombardment mass spectrometry of the cross-linked peptide . The cross-linking site is located in the highly conserved C-terminal regions of proteins S13 and S19 . In addition, the complete amino acid sequence of protein S13 from B . stearothermophilus is determined . Sequence comparison with the homologous Escherichia coli protein S13 revealed 58% identical amino acid residues.

Antimicrob Agents Chemother, 1988 May, 32(5), 642 - 5
In vitro susceptibility of Bacillus spp . to selected antimicrobial agents; Weber DJ et al.; Although often dismissed as contaminants when isolated from blood cultures, Bacillus spp . are increasingly recognized as capable of causing serious systemic infections . As part of a clinical-microbiological study, 89 strains of Bacillus spp . isolated from clinical blood cultures between 1981 and 1985 had their species determined and were tested for antimicrobial agent susceptibility to 18 antibiotics . Species of isolates were determined by the API 50CH and API 20E systems . Bacillus cereus (54 strains) was the most common species isolated, followed by B . megaterium (13 strains), B . polymyxa (5 strains), B . pumilus (4 strains), B . subtilis (4 strains), B . circulans (3 strains), B . amyloliquefaciens (2 strains), B . licheniformis (1 strain), and Bacillus spp . (3 strains) . Microdilution MIC susceptibility tests revealed all B . cereus strains to be susceptible to imipenem, vancomycin, chloramphenicol, gentamicin, and ciprofloxacin . Non-B . cereus strains were most susceptible to imipenem, vancomycin, LY146032, and ciprofloxacin . Disk susceptibility testing suggested that B . cereus was rarely susceptible to penicillins, semisynthetic penicillins, or cephalosporins with the exception of mezlocillin . In contrast, many non-B . cereus strains were susceptible to penicillins, semisynthetic penicillins, and cephalosporins, but marked variability was noted among species.

J Clin Microbiol, 1988 May, 26(5), 1061 - 2
Endocarditis caused by Capnocytophaga ochracea; Buu-Hoi AY et al.; Capnocytophaga ochracea is a gram-negative, fusiform bacillus which is part of the normal human oral flora . This organism is often isolated from periodontal lesions of patients with periodontitis and is associated with sepsis in granulocytopenic patients . We report here a case of endocarditis caused by C . ochracea.

J Urol, 1988 May, 139(5), 941 - 4
Prognostic factors in patients treated with intravesical bacillus Calmette-Guerin for superficial bladder cancer; Torrence RJ et al.; Several factors were evaluated for prognostic significance in 104 patients with a history of recurrent superficial bladder cancer treated with a 6-week course of intravesical bacillus Calmette-Guerin . Purified protein derivative skin test reactivity, tumor stage, tumor grade and number of previous tumor recurrences were evaluated in all patients . In addition, the prognostic value of a granulomatous response in the bladder was evaluated in 62 of the 104 patients . A significant correlation was reconfirmed between purified protein derivative reactivity and status free of tumor (p equals 0.041) after additional followup on 62 patients from a previous report . A significant correlation also was observed in the total patient population (p equals 0.054) . Over-all, 60 per cent of the 62 patients and 52 per cent of the 104 patients whose purified protein derivative test converted from negative to positive remained free of tumor, compared to only 28 per cent of the 62 patients and 28 per cent of the 104 whose test failed to convert to positive . Mean followup was 29.3 +/- 5.7 months in the 62 patient subgroup and 23.5 +/- 5.8 months in the total 104 patients . Bladder granuloma data were available only for the 62 patient subgroup in the previous report . With extended followup, the significant correlation previously reported between status free of tumor and granulomatous response on bladder biopsy was lost . Over-all, 29 of 37 patients (51 per cent) with granulomas compared to 8 of 25 (32 per cent) without granulomas remained free of tumor (p equals 0.132) . Tumor stage and grade, and number of previous tumor recurrences failed to show a significant correlation to status free of tumor . These results show that with extended followup, granulomatous response in the bladder lost its statistical correlation with status free of tumor, while a significant correlation was maintained for purified protein derivative responsiveness . This level of statistical significance was borderline and the purified protein derivative skin test response should not be considered useful as a prognostic indicator in individual patients.

J Bacteriol, 1988 May, 170(5), 2045 - 50
Sequence analysis of the mosquitocidal toxin genes encoding 51.4- and 41.9-kilodalton proteins from Bacillus sphaericus 2362 and 2297; Baumann L et al.; The nucleotide sequences of a 3,479-base-pair HindIII DNA fragment from Bacillus sphaericus 2362 and a 2,940-base-pair fragment from strain 2297 were determined; only minor differences were detected between them . Each contained two open reading frames coding for proteins of 51.4 and 41.9 kilodaltons . Both proteins were required for toxicity to larvae of the mosquito Culex pipiens.

J Pediatr Orthop, 1988 May-Jun, 8(3), 333 - 7
Osteitis caused by BCG vaccination; Marik I et al.; A survey of 26 Czechoslovakian children diagnosed with BCG osteitis during 1981-1986 is presented . Mycobacterial culture was attempted in 19 cases with confirmation of bacillus Calmette-Guerin (BCG) Mycobacterium bovis strain in nine cases . Symptoms appeared approximately 17 months after vaccination; the proximal tibial end, distal femur, and proximal humerus were most affected . Although vaccination has been obligatory since 1953, a different vaccine was introduced in 1980, which led to the diagnosis of BCG osteitis in 1981 . The vaccination doses, symptomatology, and methods of treatment are described . The risk of complications and a project for vaccination at later age are discussed.

Arch Biochem Biophys, 1988 May 1, 262(2), 409 - 15
Glycyl-tRNA synthetase of Escherichia coli: immunological homology with phenylalanyl-tRNA synthetase; Nagel GM et al.; Antibodies to Escherichia coli glycyl-tRNA synthetase (GlyRS) cross-react extensively with E . coli phenylalanyl-tRNA synthetase (PheRS) . These data indicate that structural homology exists between these two enzymes, the only two aminoacyl-tRNA synthetases in E . coli having an alpha 2 beta 2 subunit structure . Although only limited similarities are found in the protein sequences deduced from their known gene sequences, the presence of common epitopes in GlyRS and PheRS adds to a rather long list of physical and chemical similarities between those proteins . In addition, antibodies directed at the alpha- and beta-subunits of GlyRS inhibit both GlyRS and PheRS in the same relative manner, indicating that the function as well as the structure of subunits is similar in each enzyme . In contrast, GlyRS antibodies did not cross-react with a number of other aminoacyl-tRNA synthetase activities from E . coli, yeast, or Bacillus.

Int J Food Microbiol, 1988 May, 6(3), 259 - 61
Study of the Bacillus flora of Nigerian spices; Antai SP; Bacteriological examination of 230 samples of five different unprocessed spices (aligator pepper, red pepper, black pepper, thyme and curry powder) collected randomly from Port Harcourt main markets revealed that the spices were highly contaminated, with bacterial counts ranging from 1.8 x 10(4) to 1.1 x 10(8) per gram . Bacillus cereus was isolated in high numbers in the majority of the 230 samples examined . It was also observed that other Bacillus spp . including B . subtilis, B . polymyxa and B . coagulans occurred in significant numbers.

J Immunol, 1988 May 1, 140(9), 2994 - 9
Sensitization and desensitization to lethal effects of tumor necrosis factor and IL-1; Wallach D et al.; BALB/c mice were sensitized to lethal effects of human rTNF-alpha and of human rIL-1 alpha by simultaneous treatment with sublethal doses of actinomycin D (Act D) or D-galactosamine (GalN) . In contrast, treatment with sublethal doses of TNF or IL-1 themselves resulted in desensitization of the mice to the lethal effect of these cytokines: mice injected with TNF or IL-1 in the absence of Act D or GalN responded to a second injection of TNF or IL-1, this time together with Act D or GalN, by a significantly delayed death, or even survived . Desensitization developed rapidly (0.5-1.0 h) and abated 24 to 48 h postinjection . Each of the two cytokines induced hyporesponsiveness to its own lethal effect as well as to that of the other . Injection of TNF or IL-1 at sublethal doses resulted also in hyporesponsiveness to the lethal effect of LPS on mice primed with bacillus Calmette-Guerin, an effect which most likely is mediated by TNF and IL-1 produced in those mice in response to the LPS . TNF and IL-1 in combination had an additive effect both in lethality and in desensitization of the mice . These findings suggest that some of the deleterious effects of TNF and IL-1 are modulated by antagonistic mechanisms; mechanisms which can be suppressed by sensitizing agents, specifically by agents inhibiting the synthesis of RNA or protein; but which, in the absence of such agents, are found to be augmented in response to TNF and IL-1, thus resulting in desensitization.

J Cell Sci, 1988 May, 90 ( Pt 1), 131 - 44
The initial stages in the action of an insecticidal delta-endotoxin of Bacillus thuringiensis var . israelensis on the epithelial cells of the malpighian tubules of the insect, Rhodnius prolixus; Maddrell SH et al.; The effects of the 27 X 10(3) Mr insecticidal delta-endotoxin from Bacillus thuringiensis var . israelensis have been studied using, as a model system, isolated insect Malpighian tubules . At all concentrations of the toxin higher than 1 microgram ml-1 (4 X 10(-8) moll-1) applied to the outer surface of the tubules, fluid secretion failed within about 30 min . Except at very high concentrations, where failure always takes at least 30 s, there was an inverse relationship between the concentration of toxin and the time of failure of toxin-treated tubules . During exposure to toxin, the tubules were initially unaffected for a relatively long period and then rapid failure occurred . If the tubules were removed into toxin-free saline just before failure would have occurred, fluid secretion remained normal for at least 2 h, but on return to the origin toxin-containing saline failure was almost immediate . The toxin was found not to bind to the basement membrane . Ultrastructural changes became evident as tubule failure occurred . These initially involved modifications to the basal side of the cells, but later also to the luminal microvilli . Intercellular junctions became disassociated and cytoplasmic vacuolization occurred . The population of intramembranous particles in the basal membranes became reduced with time . Our findings suggest the following hypothesis for the initial stages in the interaction of the toxin with the tubules . Toxin molecules attach to the accessible cell membranes progressively and irreversibly . They do not readily associate by diffusing laterally in the membrane, so that toxic effects develop only when sufficiently large numbers of them attach close together . The molecules may then associate in some way as a complex, perhaps forming a pore in the membrane . Relatively few such pores lead rapidly to cell failure and death.

Ann Inst Pasteur Microbiol, 1988 May-Jun, 139(3), 363 - 77
Another Bacillus sphaericus serotype harbouring strains very toxic to mosquito larvae: serotype H6; de Barjac H et al.; Ten isolates of Bacillus sphaericus from Ghana, very toxic to mosquito larvae, have been identified as belonging to serotype H6 . These isolates can be represented by the head-group strain IAB59 . They form crystals at the sporulation stage . Their larvicidal effect on Culex pipiens and Anopheles stephensi larvae is as high as that of the most toxic strains already known, e.g . 1593 and 2362 (serotype H5a,5b) and 2297 (serotype H25) . Spore-crystal extracts of all these strains contain a 43-Kd polypeptide immunologically related to the 43-Kd polypeptide from strain 2362 described by other authors.

Mikrobiologiia, 1988 May-Jun, 57(3), 503 - 6
{Effect of the culture broth on the reactivation of refractile resting forms and on spore germination in Bacillus cereus}; Pronan SV; The filtrate of the cultural broth taken at the lag phase of Bacillus cereus 504 growth stimulated the germination of endospores and the reactivation of refractile resting forms . The energy processes were shown to be stimulated in the studied anabiotic cells.

Biochimie, 1988 May, 70(5), 645 - 8
Two-dimensional crystalline sheets of Bacillus stearothermophilus 50S ribosomal subunits containing a nascent polypeptide chain; Gewitz HS et al.; Polylysine chains were synthesized on Bacillus stearothermophilus ribosomes in a poly(A)-programmed in vitro system . After separation of the ribosomal subunits by sucrose gradient centrifugation, the polylysine chains (in contrast to the polyphenylalanine chains synthesized in a poly(U) system) reproducibly remained attached to the large ribosomal subunit . It was possible to produce two-dimensional crystalline sheets from the large ribosomal subunits containing the polylysine chains . These sheets are an essential prerequisite for three-dimensional reconstruction studies aiming to show that the tunnel in the large ribosomal subunit provides a path for the nascent polypeptide chain.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1988 May, 186(2), 153 - 65
{The significance of heat activation for the testing of bioindicators on surviving microorganisms exposed to formaldehyde}; Spicher G et al.; Heat activation is a special phenomenon: After an additional heat treatment, a larger share of the bacterial spores which had been exposed to formaldehyde proves to be viable than without such heat activation . Model studies have been performed to test the effects of heat activation on the examination of bioindicators and test objects for surviving organisms . Test objects (cotton threads of 1 cm length) contaminated with spores of Bacillus stearothermophilus were used for these trials . The test objects were exposed to a 2% formaldehyde solution at 60 degrees C . After periods of action of 30, 45, 60 .. . and 105 min, formaldehyde adhering to the test objects was neutralized . For testing these objects for surviving organisms, they were placed into a nutrient medium and incubated for 40 days at 56 degrees C . The investigation consisted of 2 parallel test series which only differed in one single point . In one series, the test objects were incubated at 56 degrees C as soon as they had been placed into the nutrient solution . In the other series, the test objects were exposed to a temperature of 95 degrees C for 1 h (heat activation) before starting incubation . The culture tubes were checked daily to see whether signs of growth (turbidity and deposits) could be observed . The frequencies of test objects with surviving organisms depending on the period of action of formaldehyde and the period of incubation determined in this way are based on the examination of 72 test objects each . Without heat activation, the share of test objects on which surviving test organisms could be detected, increased slowly with the period of incubation . Only after 30 days the counts did not increase any more when continuing the incubation (cf . Fig . 1) . In the test series in which the spores had been subjected to heat activation before the incubation period, useful results were obtained already after 3 days . They only changed slightly when incubation was continued . Moreover, the frequency of test objects on which surviving organisms could be detected was always considerably higher than without heat activation . When the frequency of test objects with surviving organisms was plotted against the period of action of formaldehyde (cf . Fig . 2A), S-shaped curves resulted.(ABSTRACT TRUNCATED AT 400 WORDS)

Biochem J, 1988 May 1, 251(3), 803 - 7
A new spectrophotometric assay for citrate synthase and its use to assess the inhibitory effects of palmitoyl thioesters; Else AJ et al.; We have demonstrated that citrate synthase may be assayed by a simple, discontinuous, spectrophotometric procedure based on the measurement of oxaloacetate utilization with 2,4-dinitrophenylhydrazine . The assay is applicable both to the purified enzyme and to cell extracts, and has the advantage that it can be used in the presence of high concentrations of thiols and thioesters . We have used this new assay in part of our investigations into the inhibitory effects of palmitoyl thioesters on diverse citrate synthases . Both palmitoyl-CoA and palmitoyl thioglycollate inhibit citrate synthases from pig heart, Bacillus megaterium and Escherichia coli, the E . coli enzyme showing the greatest sensitivity to these effectors . With palmitoyl-CoA the extent of inhibition is time-dependent, but the enzymes can be protected from the effect by the substrates oxaloacetate and acetyl-CoA . Using the dinitrophenylhydrazine assay, we have shown that the thioester bond is essential for inhibition; that is, if the palmitoyl thioesters are cleaved to give a mixture of palmitate and a thiol compound, the inhibitions of pig heart and B . megaterium citrate synthases are eliminated and that of the E . coli enzyme is markedly decreased.

Eur J Immunol, 1988 May, 18(5), 691 - 6
The processing and presentation of mycobacterial antigens by human monocytes; Bhardwaj V et al.; The processing and presentation of whole irradiated Mycobacterium tuberculosis (Mtb gamma) and its purified protein derivative (PPD) by the peripheral blood monocytes from healthy Bacillus Calmette Guerin (BCG)-vaccinated individuals was investigated . To study processing and presentation as events distinct from T cell recognition and proliferation, monocytes were pulsed with antigens for varying time intervals and fixed . The kinetics of presentation indicate that up to 2 h was required for effective presentation of PPD and 2-4 h for Mtb gamma, and that the ability to activate T cells declined as the time interval for which pulsing occurred was increased, so that responses were abolished by 8-10 h . Prefixed monocytes could not present Mtb gamma and PPD to T cells indicating that processing was an essential requisite . Lysosomotropic agents chloroquine, monensin, and leupeptin inhibited the presentation of these antigens suggesting the role of lysosomes/endosomes in processing . Furthermore, monocytes incubated with optimal concentration of antigens for different lengths of time released determinants which were still antigenic but circumvented the need for any further processing . Addition of nonprimed syngeneic monocytes, both untreated or paraformaldehyde fixed to cells which had been pulsed and fixed, restored the responses even at the later time periods when responses were not detected . This second interaction of the monocyte with T cells was not major histocompatibility complex restricted in that the addition of monocytes from another donor was equally effective.

Chest, 1988 May, 93(5), 922 - 5
T lymphocyte responses to mycobacterial antigen in AIDS patients with disseminated Mycobacterium avium-Mycobacterium intracellulare infection; Murray HW et al.; Patients with acquired immunodeficiency syndrome (AIDS) who have Mycobacterium avium-Mycobacterium intracellulare (MAI) infection typically have widely disseminated disease, often fail to respond to multi-drug chemotherapeutic regimens, and show little or no inflammatory tissue response . To determine if this clinicopathologic state correlates with in vitro lymphocyte responses to specific antigen, peripheral blood mononuclear cells from 18 patients with AIDS who had MAI bacillemia were stimulated with either particulate (heat-killed bacille Calmette Guerin {BCG}) or soluble (M intracellulare) mycobacterial antigens . In comparison to reactive cells from healthy control subjects testing positive with purified protein derivative of tuberculin (PPD) or from MAI-colonized (non-AIDS) control subjects, cells from 16 (89 percent) patients with AIDS essentially failed to show any antigen-induced proliferative activity or secretion of gamma-interferon; however, in two patients, antigen-stimulated proliferation of gamma-interferon production was modest but within the range of responses of normal healthy control subjects . Thus, although an occasional patient with AIDS can develop disseminated MAI infection despite the presence of antigen-reactive cells in vitro, most MAI-infected patients with AIDS display a striking defect in responsiveness to both particulate and soluble mycobacterial antigens . Since treatment with gamma-interferon activates the mononuclear phagocyte in vivo, these results suggest a rationale for a trial of gamma-interferon therapy in patients with AIDS who have disseminated MAI infection.

Biol Chem Hoppe Seyler, 1988 May, 369 Suppl, 315 - 22
An aspartic proteinase of erythrocyte membranes . Proposed mechanism for activation and further molecular properties; Yamamoto K et al.; An aspartic proteinase associated with human erythrocyte membranes was shown to be responsible for autodegradation of the membrane proteins at pH values below 5.0 . When the membrane was treated with phospholipase C (Bacillus cereus) or trypsin, and simply heated at 40 degrees C, the membrane-bound latent enzyme was activated, with this being accompanied by dissociation of the enzyme from the membrane . Divalent cations such as Ca2+ and Mg2+ had an inhibitory effect on the dissociation of the membrane-bound enzyme when preincubated with the membrane . The results indicate that the activation of the membrane-bound enzyme is due probably to perturbation of the normal membrane organization . When the purified enzyme was treated with 10mM 2-mercaptoethanol at 37 degrees C, the enzyme (79-82 kDa) was converted to a low molecular mass form with 42-47 kDa without any loss of activity . With the exception of treatments by thiol-reducing reagents, no conversion was observed by a variety of procedures such as exposure to 1 M NaCl and 0.1% sodium dodecyl sulfate, treatment with trypsin and incubation at pH 3.5 for up to 15 h, indicating that the enzyme consists of two polypeptide chains held together by disulfide bonds.

Mol Biol (Mosk), 1988 May-Jun, 22(3), 760 - 6
{Mutations in Escherichia coli pmp and htpR genes stabilize the products of foreign gene expression}; Kliachko EV et al.; The half lives of mRNA for Escherichia coli chloramphenicol-acetyltransferase, Bacillus amyloliquefaciens alpha-amylase and human leucocyte interferon were measured in E . coli cells by molecular RNA.DNA hybridization . The effect of mutation in pnp gene, coding polynucleotide phosphorylase, on the stability of these mRNA was studied . The half life of interferon mRNA increases from 25 to 90 s in the pnp mutant, resulting in an increase of interferon accumulation . The stability of interferon in E . coli cells depends on the htpR gene, controlling the heat shock response . The yields of leucocyte interferons alpha-2, alpha I-1 and fibroblast interferon beta increase ten times in htpR mutants . Thus, by using pnp and htpR mutants it is possible to enhance considerably the eukaryotic gene expression in bacterial cells.

Mikrobiologiia, 1988 May-Jun, 57(3), 394 - 7
{Effect of inhibitors and cations on the proteolytic activity of Bacillus mesentericus}; Imshenetskii AA et al.; The effect of some inhibitors and bivalent metal cations (Mn2+, Ca2+, Fe2+, Zn2+, Mg2+, Co2+ and Cu2+) on the proteolytic activity of two Bacillus mesentericus strains (strain 8 and strain 64 M-variant) was comparatively studied . The both enzymes were shown to be serine proteinases, but the proteinase of strain 64 was also a metal-dependent enzyme . Metal ions exerted no essential effect on the proteinase of strain 8 . Ca2+ and Mg2+ ions stimulated the proteinase activity of strain 64 whereas Fe2+ and Zn2+ ions inhibited it in the case of three substrates . Therefore, the two proteinases are different.

Am J Trop Med Hyg, 1988 May, 38(3), 608 - 12
Leukocyte subsets in the granulomatous response produced after inoculation with Mycobacterium leprae-BCG in lepromatous patients; Gross A et al.; Leukocyte subsets present in the granulomatous response produced after the inoculation of a mixture of Mycobacterium leprae and BCG in lepromatous leprosy patients were characterized in situ using monoclonal antibodies and an immunoperoxidase technique . The granuloma produced after M . leprae-BCG inoculation showed a distribution pattern similar to tuberculoid granulomas . T lymphocytes bearing the CD8 phenotype (T cytotoxic/suppressor) were sequestered to the periphery of the epithelioid tubercles and T helper-inducer CD4+ lymphocytes were distributed throughout the infiltrate . Langerhans cells CD1+ were increased in the epidermis, and in dermis they were localized mainly in the mantle surrounding the granuloma . Most of the dermal infiltrate produced after the inoculation or M . leprae-BCG expresses the HLA-DR antigen . Similarly, most keratinocytes were also positive to this MHC antigen . The granulomatous response to BCG was similar to the inoculation of a mixture of M . leprae-BCG, however acid-fast bacilla were still present . The inoculation of M . leprae produced a macrophage granuloma with no clearing of the bacilla which resembles the lepromatous leprosy granuloma.

Mikrobiologiia, 1988 May-Jun, 57(3), 410 - 4
{Polymyxin biosynthesis by Bacillus polymyxa during growth limitation by the nutritional sources}; Lykov VV et al.; The synthesis of the antibiotic polymyxin M was studied under the conditions of batch and continuous cultivation of Bacillus polymyxa var . Ross whose growth was limited with glucose, phosphate and ammonium nitrogen . Polymyxin M was synthesized when the culture growth decelerated as a result of its limitation with the above compounds . Different amounts of the antibiotic were synthesized depending on the type of a limiting factor . The highest productiveness was found in the case of glucose limitation . The optimal conditions for polymyxin M synthesis were established under the conditions of one-step continuous cultivation.

EMBO J, 1988 May, 7(5), 1515 - 26
Structural and functional analysis of Tn4430: identification of an integrase-like protein involved in the co-integrate-resolution process; Mahillon J et al.; The 4149-bp transposon Tn4430 from Bacillus thuringiensis is delineated by 38-bp inverted repeats and codes for a 113-kd protein that shares homology with the transposases (TnpA) of Tn3, Tn21 and Tn501 . Through transpositional recombination, this protein generates the formation of co-integrates between both donor and target replicons, with duplication of Tn4430 molecules . These features are characteristic of transposons of the Tn3 family (class II elements) . The second step of the transposition process, the co-integrate resolution, is mediated by a 32-kd protein . This protein (TnpI) displays regional similarities with site-specific recombinases of the integrase family, such as Int of bacteriophage lambda, Cre of bacteriophage P1 or TnpA and TnpB of the Tn554 transposon . Moreover, the 250-bp sequence upstream to the tnpI gene contains several structural features that are reminiscent of the attP attachment site of phage lambda . This unique association between the integrase-like TnpI recombinase and the TnpA transposase qualifies Tn4430 as a member of a new group within the class II mobile genetic elements.

Mol Microbiol, 1988 May, 2(3), 393 - 404
Multiplicity of delta-endotoxin genes with different insecticidal specificities in Bacillus thuringiensis aizawai 7.29; Sanchis V et al.; The hypothesis according to which multiple and different delta-endotoxin genes could determine the host-range specificity of the lepidopteran strains of Bacillus thuringiensis is being checked in the case of strains aizawai 7.29 (serotype 7) and entomocidus 601 (serotype 6) . From these strains, several crystal protein genes, belonging to different structural types, have been isolated . One of the cloned genes that is not present in strain entomocidus 601 is duplicated in strain aizawai 7.29 . This gene belongs to a previously characterized type of crystal protein gene and encodes a protein preferentially active against Pieris brassicae . Two other genes, of presumed chromosomal location, are present in both strains and each displays a unique physical map . In both strains the two genes are in close proximity and in the same orientation . The first, which belongs to a new type of crystal protein gene, encodes a 130-140 kD protein that is not significantly active against the two insect species tested . The other new type of crystal protein gene directs the synthesis of a polypeptide preferentially active against Spodoptera littoralis.

Mikrobiologiia, 1988 May-Jun, 57(3), 421 - 5
{Isolation and study of polyadenylated RNA from Bacillus intermedius}; Romakhina ER et al.; A poly(A)+RNA fraction was isolated from the overall RNA of Bacillus intermedius using chromatography on poly(U) Sepharose and was shown to be electrophoretically heterogeneous . The presence of a polyadenylate segment was confirmed by hybridization with polyuridine . The biological activity of the poly(A)+RNA was proved by the translation in Xenopus laevis oocytes . The dynamics of poly(A)+RNA synthesis was studied in the course of B . intermedius growth and the content of poly(A)+RNA was assayed in the cells grown in different media.

Lancet, 1988 Apr 30, 1(8592), 960 - 3
Epithelioid haemangioma-like vascular proliferation in AIDS: manifestation of cat scratch disease bacillus infection?
LeBoit PE, Berger TG, Egbert BM, Yen TS, Stoler MH, Bonfiglio TA, Strauchen JA, English CK, Wear DJ.
Papular and nodular skin lesions that clinically resembled Kaposi sarcoma, but histologically showed a distinct epithelioid haemangioma-like appearance, were noted in seven patients with the acquired immunodeficiency syndrome . Clusters of bacteria that had the structure of gram-negative rods were identified within each of the vascular proliferations by electron microscopy . The bacteria did not stain with the Brown-Brenn, acid-fast, or other histochemical stains for infectious organisms, but did stain with Warthin-Starry--ie, the staining profile was that described for the cat scratch disease (CSD) bacillus . Immunoperoxidase staining, using antisera raised in rabbits against cultured CSD bacillus, showed a positive reaction with the bacterium in all five cases tested . The two surviving patients have both given histories of having been scratched by a cat . In several patients, the vascular lesions regressed after therapy with antibiotics appropriate for CSD bacillus infection.

Eur J Biochem, 1988 Apr 15, 173(2), 453 - 8
The function of galactosyl phosphorylpolyprenol in biosynthesis of lipoteichoic acid in Bacillus coagulans; Yokoyama K et al.; Incubation of UDP-{14C}galactose with membranes of Bacillus coagulans led to the formation of a radioactive glycolipid, which was tentatively characterized as beta-galactosyl phosphorylpolyprenol (Gal-P-prenol) on the basis of its chromatographic behavior and data from structural analysis of its sugar 1-phosphate moiety . The sugar moiety of {14C}Gal-P-prenol was shown to be incorporated into a membrane-bound polymer, which coincided with the diacyl form of lipoteichoic acid in its chromatographic behavior on columns of Sephacryl S-300, DEAE-Sephacel and octyl-Sepharose . Hydrogen fluoride hydrolysis of the polymer afforded an alpha-galactoside identical with Gal(alpha 1----2)Gro obtained from lipoteichoic acids . The incorporation of galactose residues from {14C}Gal-P-prenol into the polymer was greatly enhanced by exogenous lipoteichoic acids, especially of the diacyl and monoacyl forms . The optimal pH and metal concentration for the Gal-P-prenol formation, respectively, were found to be 8.4 and 10 mM (MgCl2), whereas those for the transfer of galactose from this lipid intermediate to polymer were 4.5 and 16 mM (CaCl2) . The above results lead to the conclusion that Gal-P-prenol serves as the direct galactosyl donor in the synthesis of lipoteichoic acids in B . coagulans.

Biochem Biophys Res Commun, 1988 Apr 15, 152(1), 76 - 82
Alpha-factor-directed synthesis of Bacillus stearothermophilus alpha-amylase in Saccharomyces cerevisiae; Nonato RV et al.; Promoter and leader sequence of Bacillus stearothermophilus alpha-amylase gene were removed and the gene was joined in-frame to sequences encoding the leader region of Saccharomyces cerevisiae mating pheromone alpha-factor on plasmid p69A (a hybrid of pBR322 and S . cerevisiae 2-microns plasmid) . S . cerevisiae cells were transformed with plasmids containing the hybrid genes, obtaining yeast transformants which exhibit a significant extra-cellular amylolytic activity in solid medium, but not in liquid medium . Levels of alpha-amylase activity in solid medium were found to depend on the mode of fusion of the alpha-amylase gene to the alpha-factor leader region.

J Immunol, 1988 Apr 15, 140(8), 2829 - 38
Differentiation of murine macrophages to express nonspecific cytotoxicity for tumor cells results in L-arginine-dependent inhibition of mitochondrial iron-sulfur enzymes in the macrophage effector cells; Drapier JC et al.; Previous studies show that cytotoxic activated macrophages cause a reproducible pattern of metabolic inhibition in viable tumor target cells . This includes inhibition of DNA synthesis, two oxidoreductases of the mitochondrial electron transport chain (NADH: ubiquinone oxidoreductase and succinate: ubiquinone oxidoreductase), and the citric acid cycle enzyme aconitase . This pattern of metabolic inhibition is induced by a cytotoxic activated macrophage associated biochemical pathway with L-arginine deimination activity that synthesizes L-citrulline from L-arginine and oxygenated nitrogen derivatives from the imino nitrogen removed from the guanido group of L-arginine . Here we report that macrophages activated in vivo by infection with bacillus Calmette-Guerin or in vitro by murine rIFN-gamma or murine IFN-alpha/beta (in the presence of the second signal LPS in all cases) develop inhibition of aconitase and the same two oxidoreductases of the mitochondrial electron transport chain as was documented earlier in target cells of cytotoxic activated macrophages . In addition, this pattern of metabolic inhibition which develops in cytotoxic activated macrophages is caused by the L-arginine-dependent effector mechanism . Inhibition of mitochondrial respiration by effectors of the L-arginine-dependent cytotoxicity system results in a compensatory increase in activity of the glycolytic pathway . We speculate that the pattern of metabolic inhibition induced in cytotoxic activated macrophages by the L-arginine-dependent effector system causes changes in the macrophage intracellular environment that increases resistance to certain facultative and obligate intracellular pathogens.

Nucleic Acids Res, 1988 Apr 11, 16(7), 3053 - 60
BcefI, a new type IIS restriction endonuclease; Venetianer P et al.; A new Type IIS restriction endonuclease was identified, partially purified and characterized from a Bacillus cereus subsp . fluorescens strain . The enzyme recognizes the nonpalindromic sequence ACGGC and cleaves at a distance from it . The cleavage appears to occur with a +/- 1 basepair uncertainty . Thus the cleavage and recognition site is as shown below: ACGGC(N)11-13 TGCCG(N)12-14.

Nature, 1988 Apr 7, 332(6164), 564 - 8
Dissecting the catalytic triad of a serine protease; Carter P et al.; Serine proteases are present in virtually all organisms and function both inside and outside the cell; they exist as two families, the 'trypsin-like' and the 'subtilisin-like', that have independently evolved a similar catalytic device characterized by the Ser, His, Asp triad, an oxyanion binding site, and possibly other determinants that stabilize the transition state (Fig . 1) . For Bacillus amyloliquefaciens subtilisin, these functional elements impart a total rate enhancement of at least 10(9) to 10(10) times the non-enzymatic hydrolysis of amide bonds . We have examined the catalytic importance and interplay between residues within the catalytic triad by individual or multiple replacement with alanine(s), using site-directed mutagenesis of the cloned B . amyloliquefaciens subtilisin gene . Alanine substitutions were chosen to minimize unfavourable steric contacts and to avoid imposing new charge interactions or hydrogen bonds from the substituted side chains . In contrast to the effect of mutations in residues involved in substrate binding, the mutations in the catalytic triad greatly reduce the turnover number and cause only minor effects on the Michaelis constant . Kinetic analyses of the multiple mutants demonstrate that the residues within the triad interact synergistically to accelerate amide bond hydrolysis by a factor of approximately 2 X 10(6).

Eur J Biochem, 1988 Apr 5, 173(1), 9 - 16
Common features of Bacillus thuringiensis toxins specific for Diptera and Lepidoptera; Chungjatupornchai W et al.; The complete nucleotide sequence of a cloned gene encoding a 130-kDa crystal protein of Bacillus thuringiensis (B.t.) subspecies israelensis has been determined . The recombinant protein (Bt8) was purified and shown to be a mosquito-specific toxin with a LC50 value of 43 ng/ml to third-instar larvae of Aedes aegypti . Bt8 is processed by proteases or midgut extracts of mosquito larvae into toxic fragments of 68-78 kDa . Deletion mapping indicated that the active fragment of Bt8 is localized in the N-terminal half of the protoxin molecule . The deduced amino acid sequence of Bt8 has been compared with that of Bt2, a Lepidoptera-specific toxin, previously cloned from Bacillus thuringiensis berliner . Highly homologous amino acid stretches are present in the C-terminal half of the proteins . The N-terminal parts show much less sequence homology but they display a strikingly similar distribution of hydrophilic and hydrophobic amino acids . In addition, Bt8 and Bt2 show a significant immunological cross-reaction . The data indicate that although these B.t . delta endotoxins exhibit a different insect-host specificity, they are structurally related and might use a similar mechanism to interact with insect cell membranes.

Eur J Biochem, 1988 Apr 5, 173(1), 85 - 91
Binding of the delta endotoxin from Bacillus thuringiensis to brush-border membrane vesicles of the cabbage butterfly (Pieris brassicae); Hofmann C et al.; The insecticidal delta endotoxin of Bacillus thuringiensis was labeled with iodine-125 . Brush-border membrane vesicles, prepared from the midgut epithelium of Pieris brassicae larvae, known to be highly susceptible to the toxin, and from a non-target tissue: the small intestine of rat, were examined for binding of 125I-toxin . The toxin was bound specifically only to insect vesicles . Its binding to the insect membrane system was competitively inhibited by 127I-toxin and non-iodinated toxin, whereas the binding of the 125I-toxin to the mammalian membrane system was not affected by unlabeled toxin . Vesicles of P . brassicae possess two individual binding-site populations for iodinated toxin with dissociation constants of 46 nM and 490 nM . The Hill coefficients of both sites were approximately 1 and the binding capacities were 0.2 pmol and 30 pmol/mg vesicle protein for the high and the low-affinity sites respectively . The estimation of the dissociation constant for non-iodinated toxin, using a competition experiment, revealed only one binding-site population which possessed a dissociation constant of 235 nM . It is concluded that this is the binding site for the native toxin . This site was sensitive towards treatment with proteases or mixed glycosidases . It is suggested that it is a protein or a glycoprotein.

Biokhimiia, 1988 Apr, 53(4), 609 - 12
{Extracellular ribonuclease from Bacillus thuringiensis}; Chepurnova NK et al.; The ability of the strain Bacillus thuringiensis var . subtoxicus to produce extracellular ribonuclease (ribonuclease Bt) was studied . It was found that the culture medium possesses a RNA-depolymerizing activity whose maximum is observed 4-5 hours after the beginning of the linear growth phase . A three-step chromatography of the culture extract on phosphocellulose resulted in a homogeneous enzyme with a molecular mass of 12000 Da . The enzyme showed the maximum activity towards RNA at pH 8.5, catalyzed the hydrolysis of polyribonucleotides and guanosine-2',3'-cyclophosphate . Hence, the enzyme can be related to base-nonspecific cyclizing ribonucleases showing the guanylic specificity towards nucleoside-2',3'-cyclophosphates.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1988 Apr, 268(2), 209 - 12
Identification of contaminants during primary isolation of mycobacteria in the BACTEC system with the antimicrobial supplement PACT; Salfinger M et al.; 1500 sputum specimens and bronchial washings were cultured for mycobacteria . One half of the specimen was treated with N-acetyl-L-cysteine--sodium hydroxide (3%) (NALC) and the other with sodium dodecyl (lauryl) sulfate--sodium hydroxide (1%) (SDS) . The different species of contaminants found with each pretreatment method with the BACTEC radiometric system were identified . Contamination occurred in 6% by using SDS and in 10% by using NALC . The SDS method was more effective against Bacillus ssp . and Streptomyces ssp., the major contaminants . However, the growth of Pseudomonas ssp . was a problem in both methods.

Lab Anim, 1988 Apr, 22(2), 109 - 16
The microbial environment and intestinal nematode infections of Heligmosomoides polygyrus in laboratory mice; Lewis JW et al.; The difficulty of establishing primary infections of Heligmosomoides polygyrus (= Nematospiroides dubius) in ASH/CSI mice in the Laboratory Animal House at Royal Holloway and Bedford New College during a recent autumn and spring period was associated with a syndrome of worm distortion, together with zero or low worm establishment and reduced fecundity (eggs/female worm) . The eggs produced were non-viable and the egg capsule comprised a rumpled lipid and ruptured chitin layer . The egg size and peaks of egg production were also reduced and the total egg output ceased entirely by day 28 post-infection in male mice . The syndrome was repeated when control LACA mice harbouring 'normal' infections of H . polygyrus were housed on the same source of peat bedding material as the ASH/CSI mice . An increase in H . polygyrus egg production in ASH/CSI mice, removed from the peat or treated with 0.04% oxytetracycline hydrochloride suggested that the cause of the syndrome was microbial in origin . A microbiological assay of the peat, which was the common denominator of all syndrome infections, revealed an abundance of chitinase secreting species of bacteria (Bacillaceae) . Bacterial chitinase was therefore likely to rupture the chitin layer of the egg capsule producing nonviable eggs and either abnormal or no larvae . Preliminary in vitro studies using chitinase from Streptomyces griseus indicated that the hatching success of eggs of H . polygyrus was reduced as the concentration of chitinase increased.

J Wildl Dis, 1988 Apr, 24(2), 352 - 5
Serum antibody prevalence for Herpesvirus sylvilagus, Bacillus piliformis and California serogroup arboviruses in cottontail rabbits from Pennsylvania; Dressler RL et al.; A serologic survey of 60 eastern cottontail rabbits (Sylvilagus floridanus) from three counties in Pennsylvania was conducted in March 1983 . Serum antibody prevalences for Herpesvirus sylvilagus and La Crosse virus (California serogroup) were less than 4% . There was no evidence of previous exposure to either Jamestown Canyon or snowshoe hare viruses (California serogroup) . Antibody to trivittatus virus (California serogroup) was found in 60% of the 20 cottontails from York County . No cottontails had antibodies to Bacillus piliformis, the etiologic agent of Tyzzer's disease.

Arch Biochem Biophys, 1988 Apr, 262(1), 19 - 26
Leucine transport system in a facultatively alkalophilic Bacillus; Wakabayashi K et al.; Some characterizations of the leucine transport system in a facultative alkalophile, which is able to grow over a wide pH range from 7.0 to 10.5, were attempted . Although the direction of a transmembrane pH gradient of the bacterium below pH 8.2 is opposite to that above pH 8.2 (N . Koyama and Y . Nosoh (1985) Biochim . Biophys . Acta 812, 206-212), leucine transport is likely to be driven only by sodium electrochemical potential irrespective of the external pH . It was suggested that histidine and sulfhydryl groups in the leucine transporter are involved in the translocation mechanism and the pK value of the histidine residue involved is approximately 7.0.

J Manipulative Physiol Ther, 1988 Apr, 11(2), 94 - 7
Efficacy of various methods of sterilization of acupuncture needles; Sisco V et al.; The iatrogenic transmission of hepatitis B virus by inadequately sterilized acupuncture needles recently has been reported . Because some licensed chiropractors use acupuncture as a therapeutic modality, we have evaluated sterilization methods for these needles, which would be adaptable for use in a chiropractic office . Dry heat, boiling water, pressurized steam, sodium hypochlorite, and 70% alcohol were compared with a glass bead dry heat sterilizer originally developed for dental instruments . Presterilized acupuncture needles were contaminated with Bacillus stearothermophilus, Escherichia coli or Staphylococcus epidermidis and sterilized for intervals ranging from 5 sec to 30 min . The needles were then cultured to determine the efficacy of the sterilization regimen . Seventy percent alcohol was ineffective as a sterilization method . In terms of both time and convenience, the glass bead apparatus was the most efficient of the remaining methods tested . B . stearothermophilus-contaminated acupuncture needles were sterilized within 10 sec of exposure to preheated glass beads . Less than 10 sec exposure killed E . coli and S . epidermidis . A significant advantage of the glass bead sterilizer over the other methods was the absence of physical damage to the needles.

Urology, 1988 Apr, 31(4), 287 - 93
Superficial bladder cancer treated with intravesical bacillus Calmette-Guérin or adriamycin: follow-up report; Khanna OP et al.; We evaluated 139 patients with superficial bladder cancer (Stages Ta, Tl, and TIS) and treated them with either intravesical bacillus Calmette-Guerin, Tice strain (BCG), or doxorubicin hydrochloride (Adriamycin {ADR}) in a nonrandomized, multicenter study . Our follow-up study comprises 135 of these patients . Of these patients, 78 tumors were completely resected, and 61 were incompletely resected . When a proportional-hazards model (Cox) was applied, there was a statistically significant difference between the recurrence rates for the two drugs . On the basis of recurrence rates per 100 patient-months, both BCG (1.2) and ADR (0.9) worked well with completely resected tumors . However, for incomplete resections, the recurrence rate for BCG (0.9) was less than half that for ADR (1.9) . The overall recurrence rates were 1.1 and 1.3 for BCG and ADR, respectively . There have been 42 failures of treatment with either BCG or ADR . We defined failure as any recurrence of tumor; progression of the cancer in stage, grade, tumor number or size; or any residual tumor after 18 treatments (14 months of therapy) . As to the failures in patients whom we followed up, and whose treatment was either switched from ADR to BCG or continued on further BCG treatment, 53 per cent have achieved complete remission . Complete remission for BCG and ADR were 76 per cent and 52 per cent, respectively . Of the various factors considered in the study, only tumor grade and treatment drug were statistically significant . The cystectomy rate was 1 per cent for BCG-treated patients and 0 for ADR-treated patients.

J Biochem (Tokyo), 1988 Apr, 103(4), 622 - 8
Thermostable dipeptidase from Bacillus stearothermophilus: its purification, characterization, and comparison with aminoacylase; Cho HY et al.; Dipeptidase (dipeptide hydrolase {EC 3.4.13.11}) has been purified to homogeneity and crystallized from the cell extract of Bacillus stearothermophilus IFO 12983 . The enzyme has a molecular weight of about 86,000, and is composed of two subunits identical in molecular weight (43,000) . The enzyme contains 2 g atoms of zinc per mol of protein . A variety of dipeptides consisting of glycine or only L-amino acids serve as substrates of the enzyme; Km and Vmax values for L-valyl-L-alanine are 0.5 mM and 68.0 units/mg protein, respectively . The enzyme is significantly stable not only at high temperatures but also on treatment with protein denaturants such as urea and guanidine hydrochloride . The enzyme also catalyzes hydrolysis of several N-acylamino acids with Vmax values 3-30% of those for the hydrolysis of dipeptides . The thermostable dipeptidase shares various properties with bacterial aminoacylase {EC 3.5.1.14}: their subunit molecular weight, metal content and requirement, amino acid composition, and amino acid sequence in the N-terminal region are very similar.

Med Trop (Mars), 1988 Apr-Jun, 48(2), 107 - 10
{Mycobacteria encountered in Djibouti}; Auregan G et al.; The authors studied 198 strains of mycobacteriae from different sources, but mainly from bronchial secretion and adenoid pus or poundings . Because the preponderance of peripheral adenoid tuberculosis, we might fear a focus of Mycobacterium bovis . But this study gives leave to state positively that Mycobacterium tuberculosis hominis is almost exclusively the responsible and consequent by that there is an human reservoir almost exclusive . On the other hand, study of primary and secondary resistances to anti-bacillary drugs leads to express some concern particularly vis a vis modern treatments utilized as a matter of routine.

Appl Environ Microbiol, 1988 Apr, 54(4), 923 - 8
High-resolution solid-state 13C nuclear magnetic resonance of bacterial spores: identification of the alpha-carbon signal of dipicolinic acid; Lundin RE et al.; Natural-abundance solid-state 13C nuclear magnetic resonance spectra were obtained for bacterial spores for the first time by using the technique of cross-polarization magic-angle-spinning nuclear magnetic resonance spectroscopy . A resonance at about 150 ppm, detectable in spore samples having a Mn content of less than 0.05%, was consistent with an identification as the alpha-carbon signal of calcium dipicolinate; this signal was missing from a spore sample treated with acid to release dipicolinate and from a spore coat preparation . Carbohydrate peaks were particularly intense in spores and coat preparations of Bacillus macerans . Signals ascribable to beta-hydroxybutyrate were prominent in a B . cereus sample.

Protein Eng, 1988 Apr, 2(1), 45 - 8
Use of site-directed mutagenesis to probe the role of Cys149 in the formation of charge-transfer transition in glyceraldehyde-3-phosphate dehydrogenase; Mougin A et al.; Oligonucleotide-directed mutagenesis was employed to produce mutants of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of Escherichia coli and Bacillus stearothermophilus . Three different mutants proteins--His176----Asn, Cys149----Ser, Cys149----Gly--were isolated from one or both of the enzymes . The study of the properties of these mutants has shown that Cys149 is clearly responsible for the information of a charge-transfer transition, named the Racker band, observed during the NAD+ binding to apoGAPDH . This result excludes a similarity between the Racker band and the charge-transfer transition observed following the alkylation of GAPDH by 3-chloroacetyl pyridine-adenine dinucleotide.

Indian J Lepr, 1988 Apr, 60(2), 280 - 4
Involvement of palate and cheek in leprosy; Kumar B et al.; 22 Cases of bacillary positive leprosy with no apparent lesion in the oral cavity, soft or hard palate were studied for any evidence of pathological involvement . Granulomata were present in 11 (65%) out of 17 cheek biopsies studied . M . leprae were identified in four specimens only . 9 specimens (64%) out of 14 palate biopsies showed definite granulomata . M . leprae were seen in six specimens.

J Biochem Biophys Methods, 1988 Apr, 15(6), 307 - 18
Determination of the catalytic activity of cyclomaltodextrin glucanotransferase by maltotriose-methylorange assay; Makela MJ et al.; A kinetic assay of cyclomaltodextrin glucanotransferase (EC 2.4.1.19), based on the use of methylorange dye as the indicator of cyclodextrins, was studied and the reaction verified with independent HPLC analyses . The assay was optimized for the enzyme of an alkalophilic Bacillus sp . (ATCC 21783) . Described enzymological data provide a reasonable background for detection of possible errors . Numerical correlations with previous assays are presented . This method can be adopted as a standard analysis of any cyclomaltodextrin-forming enzyme.

J Neurochem, 1988 Apr, 50(4), 1158 - 63
Acetylcholinesterases from Musca domestica and Drosophila melanogaster brain are linked to membranes by a glycophospholipid anchor sensitive to an endogenous phospholipase; Fournier D et al.; The sensitivity of acetylcholinesterases (AChEs) from Musca domestica and from Drosophila melanogaster to the phosphatidylinositol-specific phospholipase C from Bacillus cereus and to the glycosylphosphatidylinositol-specific phospholipase C from Trypanosoma brucei was investigated . B . cereus phospholipase C solubilizes membrane-bound AChE, and both phospholipases convert amphiphilic AChEs into hydrophilic forms of the enzyme . The lipases uncover an immunological determinant that is found on other glycosylphosphatidylinositol-anchored membrane proteins after the same treatment . This immunological determinant is also present on the native hydrophilic form of AChE . The polypeptide bearing the active site of the membrane-bound enzyme migrates faster during sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the same polypeptide from the soluble enzyme . We conclude that AChE from insect brain is attached to membranes via a glycophospholipid anchor . This anchor is covalently linked to the polypeptide bearing the active esterase site of the enzyme and can be cleaved by an endogenous lipase.

Gene, 1988 Mar 31, 63(2), 337 - 41
Cloning and nucleotide sequencing of phenylalanine dehydrogenase gene of Bacillus sphaericus; Okazaki N et al.; The gene coding for phenylalanine dehydrogenase {PDH; L-phenylalanine: NAD+ oxidoreductase (deaminating); EC 1.4.1.-} from Bacillus sphaericus SCRC-79a was cloned onto plasmid pUC9, and the nucleotide sequence of the 2-kb DNA region of the insert was determined . A 1143-bp open reading frame consisting of 381 codons was identified as a pdh gene coding for PDH.

FEBS Lett, 1988 Mar 28, 230(1-2), 105 - 8
Interaction between the delta-endotoxin produced by Bacillus thuringiensis ssp . entomocidus and liposomes; Yunovitz H et al.; The delta-endotoxin produced by Bacillus thuringiensis ssp . entomocidus induced the release of encapsulated {14C}sucrose from reverse-phase vesicles composed of phosphatidylcholine and cholesterol . No such release was detected when the phospholipid component of the vesicles was either phosphatidylethanolamine, phosphatidylglycerol, or sphingomyelin . The toxin-induced release was competitively inhibited by negatively charged organic ions while positively charged organic ions, apart from choline chloride, had no such effect . The existence of a polar head group in the phospholipid as well as intermolecular hydrogen bonding at the membrane surface, was found to be of major importance in the toxin-liposome interaction.

Biochemistry, 1988 Mar 22, 27(6), 2229 - 34
Investigation of the Bacillus cereus phosphonoacetaldehyde hydrolase . Evidence for a Schiff base mechanism and sequence analysis of an active-site peptide containing the catalytic lysine residue; Olsen DB et al.; Reaction of Bacillus cereus phosphonoacetaldehyde hydrolase (phosphonatase) with phosphonoacetaldehyde or acetaldehyde in the presence of NaBH4 resulted in complete loss of enzymatic activity . Treatment of phosphonatase with NaBH4 in the absence of substrate or product had no effect on catalysis . Inactivation of phosphonatase with {3H}NaBH4 and phosphonoacetaldehyde, NaBH4 and {14C}acetaldehyde, or NaBH4 and {2-3H}phosphonoacetaldehyde produced in each instance radiolabeled enzyme . The nature of the covalent modification was investigated by digesting the radiolabeled enzyme preparations with trypsin and by separating the tryptic peptides with HPLC . Analysis of the peptide fractions revealed that incorporation of the 3H- or 14C-radiolabel into the protein was reasonably selective for an amino acid residue found in a peptide fragment observed in each of the three trypsin digests . Sequence analysis of the 3H-labeled peptide fragment isolated from the digest of the {2-3H}phosphonoacetaldehyde/NaBH4-treated enzyme identified N epsilon-ethyllysine as the radiolabeled amino acid . The ability of the phosphonatase competitive inhibitor (Ki = 230 +/- 20 microM) acetonylphosphonate to protect the enzyme from phosphonoacetaldehyde/NaBH4-induced inactivation suggested that the reactive lysine residue is located in the enzyme active site . Comparison of the relative effectiveness of phosphonoacetaldehyde and acetaldehyde as phosphonatase inactivators showed that the N-ethyllysine imine that is reduced by the NaBH4 is derived from the corresponding N-(phosphonoethyl) imine . On the basis of these findings, a catalytic mechanism for for phosphonatase is proposed in which phosphonoacetaldehyde is activated for P-C bond cleavage by formation of a Schiff base with an active-site lysine . Accordingly, an N-ethyllsysine enamine rather than the high-energy acetaldehyde enolate anion is displaced from the phosphorus.

J Am Vet Med Assoc, 1988 Mar 15, 192(6), 791 - 2
Bacillus piliformis infection in an adult dog; Boschert KR et al.; Bacillus piliformis infection (Tyzzer's disease) was diagnosed in a 7-year-old spayed dog that had icterus, hepatosplenomegaly, and polyuria . Hematology revealed regenerative anemia, leukocytosis, lymphopenia, and thrombocytopenia . Serum chemical analyses indicated hypocalcemia, high alkaline phosphatase activity, hypoalbuminemia, and hyperglobulinemia . At necropsy, the liver was stippled with gray-white focal lesions . Microscopically, the liver lesions were necrotic and inflammatory . Warthin-Starry-stained sections revealed rod-shaped bacteria in crisscrossing patterns characteristic of B piliformis . This dog was considerably older than dogs previously reported to have Tyzzer's disease and had a concurrent systemic hyphomycosis, suggesting it had been immunocompromised.

Biochem Biophys Res Commun, 1988 Mar 15, 151(2), 926 - 31
Sequence of the genes for the beta and epsilon subunits of the ATP synthase of Bacillus megaterium QM B1551; Hawthorne CA et al.; Four of the genes for the subunits of the proton-translocating ATPase of Bacillus megaterium have been cloned into pBR322 . Previous studies have shown that two of these genes, for the alpha and beta subunits, can complement Escherichia coli mutants defective in the genes for those subunits (Hawthorne, C.A., and Brusilow, W.S.A . 1985 . J . Biol . Chem . 261, 5245-5248) . We report here a restriction map of the cloned region and the complete nucleotide sequence of the genes for the beta and epsilon subunits as well as the deduced amino acid sequences and molecular weights of those subunits.

Eur J Biochem, 1988 Mar 15, 172(3), 731 - 8
Comparison of the in vivo and in vitro activity of the delta-endotoxin of Bacillus thuringiensis var . morrisoni (HD-12) and two of its constituent proteins after cloning and expression in Escherichia coli; Granum PE et al.; The insecticidal crystal delta-endotoxin of Bacillus thuringiensis var . morrisoni HD-12 contains at least five polypeptides in the range 126-140 kDa . Immune blotting revealed that individual proteins in this complex share homology with a range of other B . thuringiensis delta-endotoxins . In vivo the native HD-12 crystal killed a lepidopteran larva (Pieris brassicae) and a dipteran larva (Anopheles gambiae), but not the related dipteran Aedes aegypti . In vitro the solubilized activated crystal lysed Choristoneura fumiferana cells (lepidopteran) and dipteran cells derived from Anopheles gambiae and Culex quinquefasciatus but not those from Aedes aegypti . An intragenic probe derived from a B . thuringiensis var . sotto lepidoptera-specific delta-endotoxin gene hybridized with one of six plasmids extracted from HD-12 . When cloned into pUC18 two HindIII fragments from this plasmid (pEG1 and pEG2) were shown to encode polypeptides cross-reacting with HD-12 antiserum . Escherichia coli lysates containing pEG2 were toxic in vivo to lepidoptera and diptera larvae and in vitro to a broader range of insect cell lines than the native crystal . E . coli cells containing pEG3, a subclone derived from pEG1, synthesised large amounts of a 140-kDa protein in the cytoplasm as inclusion bodies . The cytotoxicity of the protein encoded by pEG3 was restricted to C . fumiferana and A . gambiae cell lines.

Cancer Res, 1988 Mar 15, 48(6), 1671 - 5
Effect of low dose cyclophosphamide on the immune system of cancer patients: depletion of CD4+, 2H4+ suppressor-inducer T-cells; Berd D et al.; We studied peripheral blood lymphocytes (PBL) from 42 patients with metastatic melanoma undergoing treatment with cyclophosphamide (CY) plus melanoma vaccine to determine whether CY immunopotentiation could be related to depletion of T-cells that function as inducers of suppression . Every 28 days, the patients were given CY, 300 mg/m2 i.v., followed 3 days later by the intradermal injection of autologous, irradiated melanoma cells mixed with Bacillus Calmette-Guerin . PBL were separated by density gradient centrifugation and cryopreserved until needed for testing . They were stained with monoclonal antibodies directly conjugated to fluorescein isothiocyanate or phycoerythrin and analyzed by two-color flow cytometry . At no time after the initiation of CY plus vaccine were there any significant changes in the percentages of helper-inducer T-cells (CD4+), suppressor-cytotoxic T-cells (CD8+), or the subpopulation of CD8+ cells expressing Leu 15, a marker for suppressor cells . Treatment of melanoma patients with CY plus vaccine resulted in a progressive fall in the proportion of CD4+ T-cells expressing the 2H4 (CD45) antigen, which identifies inducers of suppression . The reduction of CD4+, 2H4+ T-cells did not become apparent until day 28 after the first dose of CY and reached statistical significance only on days 49 (21 days after the second dose) and 105 (21 days after the fourth dose) (mean changes +/- SE: day 49, -5.4 +/- 1.4%, P less than 0.01; day 105, -9.1 +/- 2.2%, P less than 0.01; t test for nonindependent samples) . In contrast, the proportion of CD4+ T-cells expressing the antigen 4B4 (CDw29), which are true helper cells, increased slightly, although not significantly, following the institution of CY plus vaccine (mean changes: day 49, +2.9 +/- 2.1%; day 105, +3.6 +/- 2.4%) . Similar results were obtained when absolute numbers of circulating cells, rather than percentages, were analyzed . Thus the number of CD4+, 2H4+ T-cells fell from a mean of 395,000/ml on day 0 to 309,000/ml on day 49 (P less than 0.01) to 256,000/ml on day 105 (P less than 0.05) . The absolute number of CD4+, 4B4+ cells remained unchanged at the same time points . These changes were not due to progression of metastatic disease, since a comparison of patients with progressive metastases with those who were rendered disease free by surgery showed no significant differences in the reduction of the percentage of CD4+, 2H4+ T-cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Biochemistry, 1988 Mar 8, 27(5), 1648 - 52
Differential scanning calorimetry of the irreversible thermal denaturation of thermolysin; Sanchez-Ruiz JM et al.; A differential scanning calorimetry study of the thermal denaturation of Bacillus thermoproteolyticus rokko thermolysin was carried out . The calorimetric traces were found to be irreversible and highly scan-rate dependent . The shape of the thermograms, as well as their scan-rate dependence, can be explained by assuming that the thermal denaturation takes place according to the kinetic scheme N k----D, where k is a first-order kinetic constant that changes with temperature, as given by the Arrhenius equation, N the native state, and D the unfolded state or, more probably, a final state, irreversibly arrived at from the unfolded one . On the basis of this model, the value of the rate constant as a function of temperature and the activation energy have been calculated . It is shown that the proposed model may be considered as being one particular case of that proposed by Lumry and Eyring {Lumry, R., & Eyring, H . (1954) J . Phys . Chem . 58, 110} N in equilibrium D----I, where N is the native state, D the unfolded one, and I a final state, irreversibly arrived at from D . Lastly, some comments are made on the use of the scan-rate effect on the calorimetric traces as an equilibrium criterion in differential scanning calorimetry.

Ann Inst Pasteur Microbiol, 1988 Mar-Apr, 139(2), 243 - 59
Bacillus sphaericus asporogenous mutants: morphology, protein pattern and larvicidal activity; Charles JF et al.; Asporogenous mutants from Bacillus sphaericus strains 2297 and 1593-4, blocked at different stages of the sporulation process, were isolated . Two mutants (2297 Aspo30A and 2297 Aspo34) which were blocked early in sporulation did not possess any crystalline inclusions and were poorly toxic to Culex pipiens mosquito larvae . Other mutants (2297 Aspo115, 2297 Aspo24 and 1593-4 Aspo12) which were blocked at later stages synthesized crystal-like inclusions next to the forespores, and were highly toxic to mosquito larvae . Electrophoretic protein analysis of alkali extracts from mutants and wild type strains confirmed the absence of toxic crystal-related proteins in early-blocked mutants and their presence in later ones . Western blots with antisera directed against the crystal proteins confirmed those observations.

J Assoc Off Anal Chem, 1988 Mar-Apr, 71(2), 337 - 40
Determination of residual cephalexin in chick tissues by Bacillus stearothermophilus paper disc assay; Okada J et al.; A paper disc method is described for determination of residual cephalexin (CEX) in chick tissues . A trichloroacetic acid extract of plasma and tissues is chromatographed on a macroreticular resin (Diaion HP-20) column to remove endogenous antibacterial substances interfering with the assay . The eluate is evaporated to dryness and the residue, dissolved in methanol-water (1 + 2), is subjected to a paper disc assay using Bacillus stearothermophilus var . calido-lactis C953 NIZO as a test organism . The detection limit was 0.0375 ppm in tissue; the average recovery of CEX ranged from 72.4% in skin to 90.4% in plasma . Water containing 200 or 500 mg/L of CEX was given ad libitum to 2-week-old chicks for 10 days; the highest levels of CEX were found in the kidney, and the lowest were found in muscle at 0 h of withdrawal . CEX disappeared from most tissues at 24 h after withdrawal except from skin of chicks given 500 mg/L . However, the drug was not detected in the skin at 48 h after withdrawal.

Rev Infect Dis, 1988 Mar-Apr, 10(2), 424 - 7
Septic anaerobic jugular phlebitis with pulmonary embolism: problems in management; Bach MC et al.; Anaerobic gram-negative bacillary bacteremia and multiple septic pulmonary emboli developed rapidly in two previously healthy young men after an episode of pharyngitis . One patient developed proptosis and subsequent uniocular blindness . In both cases facial swelling was an early sign of jugular vein involvement . In patients not responding to antibiotic therapy, systemic anticoagulation or surgical venous ligation may be potentially useful as an additional therapeutic measure . Septic jugular vein phlebitis is a serious condition that requires early recognition and rapid institution of appropriate therapy.

Int J Lepr Other Mycobact Dis, 1988 Mar, 56(1), 21 - 6
Facies leprosa: resorption of maxillary anterior alveolar bone and the anterior nasal spine in patients with lepromatous leprosy in Mali; Marks SC Jr et al.; Resorption of the anterior nasal spine and alveolar bone in the anterior maxilla was measured in 39 patients with lepromatous leprosy in Mali . Bone resorption occurred in both of these sites, but resorption in one did not predict resorption in the other . These data are interpreted to mean that resorptions of bone anterior (nasal spine) or inferior (alveolar bone) to bacillary populations in the nasal mucosa of patients with lepromatous disease in Mali occur independently.

J Bacteriol, 1988 Mar, 170(3), 1054 - 62
Bacillus sporulation gene spo0H codes for sigma 30 (sigma H); Dubnau E et al.; The DNA sequences of the spo0H genes from Bacillus licheniformis and B . subtilis are described, and the predicted open reading frames code for proteins of 26,097 and 25,447 daltons, respectively . The two spo0H gene products are 91% identical to one another and about 25% identical to most of the procaryotic sigma factors . The predicted proteins have a conserved 14-amino-acid sequence at their amino terminal end, typical of sigma factors . Antibodies raised against the spo0H gene product of B . licheniformis specifically react with RNA polymerase sigma factor protein, sigma 30, purified from B . subtilis . We conclude that the spo0H genes of B . licheniformis and B . subtilis code for sigma 30, now known as sigma H.

J Bacteriol, 1988 Mar, 170(3), 1034 - 40
Evidence for movement of the alpha-amylase gene into two phylogenetically distant Bacillus stearothermophilus strains; Satoh H et al.; The gene for an alpha-amylase cloned from strain DY-5 of Bacillus stearothermophilus was used to examine to what extent the corresponding genes are structurally similar in other B . stearothermophilus strains . The structure of the gene itself was almost identical in DY-5 and a group of strains represented by strain 799 . The gene was not detected at all in strain DSM2334, which was phenotypically amylase deficient . Comparison of the structure of 5S rRNA and electrophoretic pattern of the ribosomal proteins indicates that strains DY-5 and DSM2334 are closely related to each other, whereas strain 799 is phylogenetically very distant from the two . We estimate that strain 799 separated from DY-5 and DSM2334 some 420 million years ago . Nucleotide sequencing of the region containing the amylase gene from strains DY-5 and 799 revealed the presence of a 3.4-kilobase stretch that was highly similar in the two strains . Furthermore, comparison of the restriction map surrounding the amylase gene of DY-5 with that of a corresponding region in DSM2334 indicated that the former strain contained an extra segment 5.5 kilobases in length, which included the 3.4-kilobase stretch mentioned above . This segment was missing in DSM2334 . It thus appears that the alpha-amylase gene was brought into strains DY-5 and 799 from outside despite a large phylogenetic distance.

J Am Mosq Control Assoc, 1988 Mar, 4(1), 64 - 72
Laboratory study of the influence of water temperature and pH on Bacillus thuringiensis var . israelensis efficacy against black fly larvae (Diptera: Simuliidae); Lacoursiere JO et al.; An experimental formulation of Bacillus thuringiensis var . israelensis was used in the laboratory to assess the influence of water temperature and pH on the relationship between concentration, duration of exposure, and mortality of the northern black fly species Simulium decorum and Prosimulium mixtum/fuscum group . Mortality increases in both species with increases in duration of exposure, concentration, temperature and pH . Onset of death is shortened by increase in concentration and temperature . As temperature rises, the concentration of B.t.i . required to induce mortality decreases; the sharpest decline occurring between 12 and 18 degrees C for S . decorum, and between 4 and 8 degrees C for P . mixtum/fuscum larvae . Lower pH induces a loss of efficacy of the B.t.i . formulation on S . decorum larvae at 4 and 12 degrees C . Dialysis of the B.t.i . formulation at pH 11 for 2 h increases its potency against S . decorum larvae, suggesting an effect of an extralarval alkaline hydrolysis on the B.t.i . efficacy . An alkaline prehydrolysis of the paracrystalline bodies could therefore be used in cold and acidic environments to compensate for loss of efficacy.

J Am Mosq Control Assoc, 1988 Mar, 4(1), 39 - 43
Host range and selected factors influencing the mosquito larvicidal activity of the PG-14 isolate of Bacillus thuringiensis var . morrisoni; Lacey LA et al.; Laboratory bioassay of the PG-14 isolate of Bacillus thuringiensis var . morrisoni (serotype 8a:8b) against early fourth instar larvae of 8 species of mosquitoes revealed a range of susceptibilities similar to the susceptibilities of these species to Bacillus thuringiensis var . israelensis (serotype 14) . The most susceptible species were: Culex quinquefasciatus, Cx . salinarius, Anopheles albimanus and Aedes aegypti . The least susceptible species tested was An . quadrimaculatus . Separate bioassays of PG-14 against the four instars of Ae . aegypti demonstrated a strong negative correlation (R = -0.97) between larval age and susceptibility . Temperature significantly affected the stability of larvicidal toxin in aqueous suspensions of PG-14 . Larvicidal activity of a bacterial suspension was nearly completely eliminated after 132 days of storage at 31 degrees C, but was essentially unchanged for those suspensions stored at 4 degrees C.

Infection, 1988 Mar-Apr, 16(2), 86 - 90
Central venous catheter infections in pediatric patients--in a community hospital; Kumar A et al.; We reviewed the records of 23 pediatric patients who had received at least one central venous catheter during a two-year period . Nine patients had acute lymphoblastic leukemia (ALL), nine had other hematologic/oncologic diagnoses, and five had cystic fibrosis . Twenty-nine of 65 febrile episodes in 16 patients were associated with a catheter-related infection . Twenty of 40 catheters were associated with an infection over a period of 7,229 catheter days . For every 1,000 catheter days, four episodes of infections were observed . The number of infections/1,000 catheter days, the average life of a catheter (approximately equal to 180 days), and mean number of days elapsing before the first infection were not significantly different in the three diagnostic groups . Broviac catheters were used most often (24/40), followed by Quinton (9/40) and Port-a-Cath (7/40) . Broviac catheters lasted twice as long (224 days, p less than 0.01) as Quinton and Port-a-Cath . Gram-positive cocci were isolated most frequently and Staphylococcus epidermidis was the most common pathogen . No consistent relationship between an absolute neutrophil count of less than 1,000/mm3 and infection with gram-positive cocci was seen . However, seven of eight episodes of gram-negative bacillary infections occurred in patients with an absolute neutrophil count of less than 1,000/m3 (p less than 0.005) . Those patients who were not considered terminally ill responded well to antimicrobials . Catheter removal was necessary in only two instances.

J Gen Microbiol, 1988 Mar, 134 ( Pt 3), 743 - 9
An assessment of taxonomic congruence between DNA-DNA hybridization and pyrolysis gas-liquid chromatographic classifications; O'Donnell AG et al.; The existence of subgroups within Bacillus megaterium has been reported previously on the basis of DNA-DNA hybridization and DNA base composition studies . In this study the strains used to define these subgroups have been reanalysed by pyrolysis gas-liquid chromatography . The resultant two-group classification of the test strains was directly comparable with that obtained from the previous nucleic acid analyses at the between-group level . However, comparisons of the test strains at the within-group level proved less successful.

J Appl Bacteriol, 1988 Mar, 64(3), 257 - 64
Characterization of Bacillus cereus strains isolated from drugs and evaluation of their toxins; Garcia Arribas ML et al.; The microbial contamination of 68 samples of topical and 324 samples of oral medicaments has been studied . The most common group of contaminants was members of the genus Bacillus (34.4%) . Because of the pathogenic significance of B . cereus, 39 strains were characterized by morphology and biochemical properties . All except three showed most of the characteristics of the type strain . They were highly resistant to lincomycin, polymyxin B and penicillin G-cephalosporin and were susceptible to streptomycin, erythromycin and chloramphenicol . Enterotoxin, phospholipase C and haemolysin production were also studied: 33 strains gave a positive vascular permeability reaction, four of them causing necrosis, and 24 showed positive mouse lethal tests . All the strains had phospholipase activity . The majority also exhibited differing degrees of haemolysis . Permeability factor was related to mouse lethality and haemolytic activity . Phospholipase C was not related to any of the above activities.

Appl Environ Microbiol, 1988 Mar, 54(3), 699 - 702
Incidence and characterization of Bacillus cereus isolates contaminating dairy products; Wong HC et al.; A total of 293 dairy products purchased from local markets were examined to determine the incidence of and characterize Bacillus cereus . Isolations were made on mannitol-egg yolk-polymyxin B agar medium and confirmed by several staining and biochemical tests . B . cereus occurred in 17% of fermented milks, 52% of ice creams, 35% of soft ice creams, 2% of pasteurized milks and pasteurized fruit- or nut-flavored reconstituted milks, and 29% of milk powders, mostly in fruit- or nut-flavored milk mixes . The average population of B . cereus in these dairy products was 15 to 280 CFU/ml or CFU/g (range, 5 to 800) . The characteristics of these B . cereus isolates in terms of heat resistance, biochemical reactions, and antibiotic susceptibility were similar to previously reported data except for a higher utilization of sucrose . Some isolates were especially resistant to carbenicillin, nalidixic acid, streptomycin, and tetracycline . The MICs for the isolates were also determined . All of the tested isolates lysed rabbit erythrocytes; 98% showed verotoxicity, 68% showed cytotonic toxicity for CHO cells, and 3 of 11 selected isolates that showed strong hemolysin activity killed adult mice.

Jpn J Clin Oncol, 1988 Mar, 18(1), 69 - 74
Bacillus cereus bacteremia in an adult with acute leukemia; Funada H et al.; Bacillus cereus, which used to be considered non-pathogenic, was isolated from the blood of a patient with acute leukemia who was receiving intensive chemotherapy . Fatal bacteremia developed with a clinical syndrome of acute gastroenteritis, followed by both meningoencephalitis with subarachnoid hemorrhage and multiple liver abscesses probably caused by infective vasculitis . Surveillance stool cultures revealed colonization with the organism prior to the onset of diarrhea, and repetitive blood cultures were found to be positive . Thus, this case suggested some new important clinicopathologic features of true B . cereus bacteremia complicating acute leukemia.

Urology, 1988 Mar, 31(3 Suppl), 17 - 9
Intravesical chemotherapy: how effective is it?
Heney NM.
Intravesical chemotherapy has been shown to reduce or prevent recurrence of low-grade, low-stage transitional cell tumors of the bladder . Thiotepa, mitomycin C, doxorubicin, and bacillus Calmette-Guerin (BCG) are the drugs most widely used for intravesical chemotherapy in the United States; only thiotepa has FDA approval for use within the bladder . Results of studies with these agents are reviewed . Prospective, randomized studies are needed to determine the ability of the agents to prevent progression of high-grade tumors to muscle invasion.

Proc Natl Acad Sci U S A, 1988 Mar, 85(5), 1408 - 11
Phosphorylation of synthetic random polypeptides by protein kinase P and other protein-serine (threonine) kinases and stimulation or inhibition of kinase activities by microbial toxins; Abdel-Ghany M et al.; A synthetic random polymer of threonine and glutamate (1:4.4) is readily phosphorylated by protein kinase P but not by five other protein-serine (threonine) kinases . A synthetic random polymer of serine and arginine (1:3) is readily phosphorylated by protein kinase A and protein kinase C but not by protein kinase P . Although the amino acid sequences surrounding the phosphorylated serine (threonine) residue have been demonstrated in studies with small synthetic polypeptides to be decisive factors in the rate at which they are phosphorylated, the findings with the large synthetic polypeptides suggest that in the case of proteins the size, the tertiary structure, and particularly the electrostatic interactions are equally or more important contributing factors . Syringomycin, a toxin from Pseudomonas syringae, and polymyxin B, from Bacillus polymyxa, stimulate protein kinase P, strongly inhibit protein kinase C, and have no effect on protein kinase A . Basic polypeptides with high lysine content are phosphorylated by ATP nonenzymatically.

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