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