Bacillus

Gene, 1990 Mar 1, 87(1), 63 - 70
Cloning and characterization of the Bacillus sphaericus genes controlling the bioconversion of pimelate into dethiobiotin; Gloeckler R et al.; Using 8.8 kb of genetic information from Bacillus sphaericus, it was possible to confer to Escherichia coli bio- strains, including delta bioA-D, bioC-, bioH-, the ability to convert exogenous pimelate into biotin . The bio genes were borne on two recombinant plasmids with inserts of 4.3 kb and 4.5 kb, which had been isolated from a genomic bank of HindIII-digested B . sphaericus DNA, by phenotypic complementation of various E . coli bio mutants . The B . sphaericus bioD and bioA genes were unambiguously identified within the 4.3-kb insert and shown to be closely linked to bioY (coding for a protein with a presently unknown function) and to bioB {Ohsawa et al., Gene 80 (1989) 39-48} . These genes are clustered in the order bioDAYB . The 4.5-kb fragment contains genetic information for three different proteins, the products of bioX, bioW and bioF . Complementation studies using an E . coli bioF mutant and a B . subtilis bio112TG3 strain, revealed that the third ORF of this cluster encodes 7-keto-8-aminopelargonic acid synthetase . A combination of bioW and bioF allows an efficient complementation of E . coli bioC and bioH mutants, provided that pimelate is added to the biotin-depleted growth medium . No function could be identified for the product of bioX . The gene order of this cluster is bioXWF . By sequence analysis, the two cloned DNA fragments were shown to bear overlapping open reading frames and secondary structures at their 3' ends, typical of transcription terminators.(ABSTRACT TRUNCATED AT 250 WORDS)

J Assoc Off Anal Chem, 1990 Mar-Apr, 73(2), 202 - 5
Factors influencing optimization of diffusion assays for antibiotics; Brady MS et al.; Agar nutrient content, cylinder charge volume, thickness (volume) of the agar layer, and incubation temperature were 4 factors varied to determine their effect(s) on the optimization of the cylinder-plate diffusion assay . Chlortetracycline was the pilot antibiotic and Bacillus cereus was used as the assay organism . Zones of inhibition were larger when the incubation temperature was lower than that which was commonly used and/or when the nutrient level was decreased; the zones were smaller when the incubation temperature was raised and/or when an increased nutrient level was used . The thickness (volume) of the assay layer played the most important role; the thinner the layer the less the effect the cylinder charge volume had on the zone diameter . The slopes of the response lines were minimally affected by cylinder charge volume . For a 7 mL assay layer per standard Petri plate, cylinder charge volumes ranging from 150 to 250 microL had little effect on zone diameter . The linearity of the response line was unaffected by assay layer thickness (volume), nutrient level, temperature of incubation, or cylinder charge volume . As long as the conditions for the assay were standardized, there were no discernible effects on recoveries or potencies.

J Invertebr Pathol, 1990 Mar, 55(2), 272 - 7
Response of nuclear polyhedrosis virus-resistant Spodoptera frugiperda larvae to other pathogens and to chemical insecticides; Fuxa JR et al.; Selection in the laboratory for Spodoptera frugiperda (Sf) resistant to nuclear polyhedrosis virus (NPV) affected the susceptibility of the insect to certain other mortality agents, including a chemical insecticide . Median lethal concentrations (LC50S) and associated statistics were compared for several mortality agents between colonies of NPV-resistant and -susceptible (control) insects . Compared to the susceptible insects, the NPV-resistant insects were cross-resistant to the S . frugiperda granulosis virus and to the Autographa californica NPV based on nonoverlap of 95% fiducial limits of the LC50S . The NPV-resistant insects were significantly more susceptible to methyl parathion than the control insects . The two colonies of S . frugiperda did not differ significantly in their response to Bacillus thuringiensis, Vairimorpha necatrix, or carbaryl . The cross-resistance experiments were based on per os exposure of the insects to the pathogens and insecticides; the susceptibility of the resistant and control insects did not differ significantly when the Sf NPV was injected into the hemocoel or when methyl parathion was applied topically.

J Urol, 1990 Mar, 143(3), 502 - 6
Bacillus Calmette-Guerin versus doxorubicin versus thiotepa: a randomized prospective study in 202 patients with superficial bladder cancer; Martinez-Pineiro JA et al.; We report the second interim analysis of data from a randomized prospective trial comparing the prophylactic effect of 15 courses of 50 mg . doxorubicin, 50 mg . thiotepa or 150 mg . bacillus Calmette-Guerin instilled intravesically against recurrences and progression of superficial transitional cell bladder cancer . Of 202 enrolled patients 176 currently are evaluable with a mean follow-up of 3 years (range 3 to 97 months) . The number of patients with recurrences was significantly lower in the bacillus Calmette-Guerin arm (9 of 67) compared to the doxorubicin (23 of 53, p equals 0.002) and thiotepa (20 of 56, p equals 0.003) arms . The over-all recurrence index per 100 patient-months also was lower for the bacillus Calmette-Guerin versus the thiotepa and doxorubicin groups (0.53 versus 1.55 and 1.7, respectively) . Bacillus Calmette-Guerin also was superior in preventing recurrences and progression of high risk tumors, that is stage T1, grade 3 or multiple growths, associated or not with carcinoma in situ . In the stage T1 category 19 of 32 (60%) tumors recurred under treatment with doxorubicin, 11 of 33 (33%) with thiotepa and 6 of 49 (12%) with bacillus Calmette-Guerin . Toxicity to intravesical bacillus Calmette-Guerin was higher compared to the other drugs but it was not limiting: bladder irritability and malaise occurred in 42% of the patients, granulomatous cystitis in 16.4% and bladder contraction in 1.4% (1 of 64) . The latter complication occurred in a patient whose stage T1m grade 2 tumors had recurred 3 times, who underwent 3 transurethral bladder resections within 15 months and who had received thiotepa for 4 months after having been removed from the study 11 months after entry . Three patients in the doxorubicin group (5.6%) underwent radical cystectomy for local urothelial progression . One patient (1.8%) in the same group died of distant progression . Our preliminary results suggest that at the dose, periodicity and duration used in the study bacillus Calmette-Guerin is significantly superior to the chemotherapeutic agents doxorubicin and thiotepa for the prophylaxis of recurrence and retardation of progression in superficial transitional cell bladder tumors.

J Neurochem, 1990 Mar, 54(3), 1047 - 55
Effect of phospholipase C from Bacillus cereus on the release of membrane-bound choline-O-acetyltransferase from rat hippocampal tissue; Carroll PT et al.; Some of the enzyme choline-O-acetyltransferase (ChAT) associated with central cholinergic nerve terminals appears to be non-ionically associated with membranes . In the present study, we tested the possibility that some membrane-bound ChAT might be anchored to membranes by a phosphatidylinositol linkage by incubating rat hippocampal tissue with phospholipase C (PLC) from Bacillus cereus . The PLC selectively augmented the release of ChAT; also, the glycosylphosphatidylinositol-PLC inhibitor, zinc, blocked this increase in release . When control and PLC-treated hippocampal tissues were subjected to Triton X-114 phase separation, a procedure that separates amphiphilic from hydrophilic proteins, the detergent-soluble, membrane-bound fraction of tissue ChAT appeared to be the source of the ChAT released by PLC into the incubation medium . Zinc also blocked the temperature-dependent release of ChAT, but not lactic dehydrogenase, from hippocampal tissue . Extracellular membrane-bound ChAT appeared to be the source of the ChAT released by a low exogenous concentration of PLC, as well as that released by a temperature-dependent process during tissue incubation . Phosphatidylinositol-specific PLC from Bacillus thuringiensis released ChAT, but not lactic dehydrogenase, from a crude synaptosomal fraction prepared from rat hippocampal tissue . These results suggest that some of the membrane-bound ChAT in rat hippocampal tissue may be extracellular and anchored to the membrane by phosphatidylinositol, and also that an endogenous factor in hippocampal tissue may function to remove this extracellular ChAT from the membrane.

J Am Mosq Control Assoc, 1990 Mar, 6(1), 93 - 5
Efficacy of various Bacillus thuringiensis var . israelensis formulations against Psorophora columbiae larvae as assessed in small rice plots, 1984-88; Meisch MV et al.; Granular, liquid and briquette formulations of Bacillus thuringiensis var . israelensis were tested in small rice plots during 1984-88 against Psorophora columbiae larvae . Vectobac AS, Skeetal G, Teknar HPD, ABG 6172, ABG 6188, ABG 6193, ABG 6197, ABG 6199, ABG 6138F and ABG 6221 provided excellent control at high dosages and good control (generally greater than 85%) at relatively low concentrations . ABG 6221 and ABG 6172 provided excellent control at low concentrations . Additional testing of these formulations will be required to ascertain specific lower limits such that more economical larval mosquito control can be obtained.

J Invertebr Pathol, 1990 Mar, 55(2), 189 - 201
Comparison of development of Bacillus thuringiensis subsp . israelensis and Bacillus sphaericus in mosquito larvae; Pantuwatana S et al.; Immunofluorescent staining was used with thin sections of paraffin-embedded specimens to detect the development of Bacillus thuringiensis var . israelensis and Bacillus sphaericus in the gut of mosquito larvae . The third- and fourth-instar larvae of Aedes aegypti, Anopheles maculatus, and Culex quinquefasciatus were fed either vegetative cells or spores of the bacteria . Spore germination, multiplication, and sporulation were studied in the larvae of each species . The spores of B . thuringiensis var . israelensis and B . sphaericus strain 2297 could germinate and cells could sporulate in the larval body . The vegetative cells of B . sphaericus strain 810428 were also able to produce spores in the mosquito larval gut, but the germination of spores could not be detected in the larvae . Multiplication of all bacterial species was observed after the larvae died . Growth of the bacteria in distilled water containing crude extracts of larvae made from each species was compared with that in synthetic medium (nutrient broth) . They could produce spores and toxins in all the media used and the toxins had larvicidal activity against the target mosquitos Ae . aegypti, An . maculatus, and C . quinquefasciatus.

J Clin Lab Immunol, 1990 Mar, 31(3), 127 - 36
Competitive action of a biological response modifier, PSK, on a humoral immunosuppressive factor produced in tumor-bearing hosts; Matsunaga K et al.; We investigated the effect of PSK, a protein-bound polysaccharide obtained from the basidiomycetes Coriolus versicolor, on an immunosuppressive factor produced in tumor-bearing animals . Oral administration of PSK suppressed the growth of the tumor in C3H/He mice bearing X5563 plasmacytoma or MH134 hepatoma, but affected mice bearing MM102 mammary tumor little . PSK prevented the reduction in splenic lymphocyte blastogenesis caused by phytohemagglutinin that occurs in mice bearing X5563 tumors or MH134 hepatoma . The lymphocyte blastogenesis affected little by tumor or PSK in mice bearing MM102 tumors . The effect of sera on the blastogenesis of lymphocytes caused by phytohemagglutinin was different with different tumors in the C3H/He mice . Serum of mice bearing X5563 tumors inhibited blastogenesis, but serum of mice bearing MH134 hepatoma or MM102 tumors promoted it . The sera of mice bearing MH134 hepatoma contained both inhibitory and promotive factors; those of mice bearing X5563 tumors contained an inhibitory factor, and those of mice bearing MM102 tumors contained a promotive factor . The oral administration of PSK reduced the inhibition caused by the sera of mice bearing X5563 tumors . The promotive activity of sera from mice bearing MH134 hepatoma was augmented by PSK; that of sera in mice bearing MM102 tumors was not affected by PSK . Living Bacillus Calmette-Guerin did not have such effects in any of these mice . Serum immunosuppressive activity was also reduced by PSK in various tumor lines of rodents . These results suggest that PSK acts by reducing the activity of immunosuppressive factors produced in tumor-bearing hosts.

J Gen Microbiol, 1990 Mar, 136 ( Pt 3), 545 - 53
Isolation and characterization of a unique division mutant of Bacillus megaterium; Lach DA et al.; A filamentous division mutant, PV302, of Bacillus megaterium QM B1551 was isolated while screening for sporulation-defective mutants after nitrosoguanidine mutagenesis . Both phase-contrast and electron microscopy revealed that the mutant produced small spherical cells as well as filaments . It also accumulated large amounts of poly-beta-hydroxybutyrate . Poly-beta-hydroxybutyrate accounted for 16% of the dry weight of the mutant strain even after 28 h growth . In comparison to the parental strain, the division mutant also showed both an inability to sporulate and a reduced growth rate . All these phenotypes transduced together . Revertants gained the ability to sporulate, divide, and grow normally . Transductional mapping of the mutation, designated div-1, established a new linkage group for B . megaterium consisting of div-1 and the pyrimidine biosynthesis genes pyrD BCF . The spherical cells were separated from filaments by sucrose gradients and were tested for nucleic acid content and viability . The purified spherical cell fraction contained one-fifth the amount of DNA per mg protein as compared with the filamentous cell fraction and was shown to contain both non-viable minicells and some cells capable of growing after a lag of about 4 h . This suggests that the mutation not only causes defects in septum placement and sporulation, but may possibly affect DNA partitioning.

Indian J Med Res, 1990 Mar, 91, 149 - 50
Extra cellular hydrolytic enzyme secretion in Bacillus thuringiensis H14 & B . sphaericus & their significance in media design; Kuppusamy M et al.; Level of extracellular proteolytic and amylolytic enzyme production was determined to assess the ability of these strains in utilizing complex carbon and nitrogen sources . Protease secretion reached maximum at around 12th h of growth in case of all the three B . thuringiensis strains and declined sharply thereafter . In B . sphaericus strains the protease level gradually increased and reached maximum at around 24th h of growth . Amylase activity was not detectable in the culture supernatants of B . sphaericus strains whereas all the three B . thuringiensis strains tested showed significant amount of amylase activity.

J Biochem (Tokyo), 1990 Mar, 107(3), 480 - 5
A novel aco-type cytochrome-c oxidase from a facultative alkalophilic Bacillus: purification, and some molecular and enzymatic features; Qureshi MH et al.; A novel aco-type cytochrome-c oxidase was highly purified from the facultative alkalophilic bacterium, Bacillus YN-2000, grown at pH 10 . The enzyme contained 9.0 nmol heme a/mg protein . It contained 1.23 mol of protoheme, 1.06 mol of heme c, 2.0 g atoms of copper, 2.5 g atoms of iron, and 1.8 g atoms of magnesium per mol of heme a . The enzyme molecule seemed to be composed of two subunits with Mrs of 52,000 and 41,600 . On the basis of these results, the enzyme seemed to contain one molecule each of heme a, protoheme, and heme c per minimal structural unit (Mr, 93,600) . Only protoheme among the three kinds of hemes in the enzyme reacted with CO and CN- . Heme a did not react with CO; cytochrome a3 did not seem to be present in the enzyme . The enzyme oxidized 314 mol of horse ferrocytochrome c per heme a per sec at pH 6.5 and the catalytic activity was 50% inhibited by 7.65 microM KCN . The enzymatic activity was found to be optimal at pH 6.0.

J Am Mosq Control Assoc, 1990 Mar, 6(1), 135 - 7
Field trial of Abate and Teknar for Simulium (Diptera: Simuliidae) control in India; Das SC et al.; Simulium breeding was eliminated in a river up to 30 km with 0.1 ppm Abate (temephos) for 30 min exposure . Teknar (Bacillus thuringiensis var . israelensis) at 10 ppm for 10 min exposure resulted in 98.4% and 87.9% reduction up to 20 m in 2 sections of a small stream.

Lett Appl Microbiol, 1990 Mar, 10(3), 141 - 3
DNA fingerprinting of the mosquito pathogen Bacillus sphaericus with M13 DNA as a probe; Abadjieva AN et al.; Hypervariable nucleotide sequences were detected in Bacillus sphaericus by hybridization with radioactively labelled M13 DNA . Different serotypes could be distinguished by their hybridization profiles . The appearance of bands common for mosquito-pathogenic strains and their absence in an apathogenic strain opens the probability that M13 could hybridize to specific alleles, related to insect toxicity.

Agric Biol Chem, 1990 Mar, 54(3), 715 - 24
Construction of chimeric insecticidal proteins between the 130-kDa and 135-kDa proteins of Bacillus thuringiensis subsp . aizawai for analysis of structure-function relationship; Nakamura K et al.; Eight chimeric insecticidal protein (IP) genes were constructed between the 130-kDa and 135-kDa IP genes of Bacillus thuringiensis subsp . aizawai, and expressed in Escherichia coli JM103 cells . The characterization of the produced chimeric IPs indicated that the variable region (VR1) in the amino-terminal half of the IPs is responsible for the insecticidal activity against larvae of Spodoptera litura and Plutella xylostella . The carboxy-terminal half of VR1 was important for the formation of the 60-kDa active fragment in the gut juice of S . litura larvae . Also, combination of the other two variable regions (VR2 and VR3), which were in the central and carboxy-terminal portions of the IPs, appeared to be related to the solubility of the IPs in the gut juice.

Agric Biol Chem, 1990 Mar, 54(3), 641 - 8
Expression of the beta-cyclodextrin glucanotransferase gene of an alkalophilic Bacillus sp . #1011 in Escherichia coli cells and characterization of the synthesized enzyme; Kimura K et al.; To express efficiently the gene for extracellular beta-cyclodextrin glucanotransferase (beta-CGTase) of an alkalophilic Bacillus sp . #1011 using the E . coli promoters (tac, trp and PL promoters), three DNA fragments starting from the nucleotide positions +1, -18, and -48 of the translation initiation site of the gene were prepared and they were fused with the promoters . The maximum production of the enzyme, which was located mainly (90%) in the periplasm of the E . coli strain, was observed in the combination of the trp promoter and the beta-CGTase gene starting from the -48 nucleotide position in the presence of the inducer, IAA . The production of the enzyme was increased to 5.5 times that by the E . coli harboring the original plasmid and to approximately 3 times higher than the extracellular production of the enzyme by the parental Bacillus sp . #1011 . The properties including the stability and optimum in the high pH range (pH 9) of the extracellular beta-CGTase from the alkalophilic Bacillus was conserved in the periplasmic enzymes of the E . coli cells.

Biochim Biophys Acta, 1990 Feb 26, 1033(2), 184 - 8
Variation of the surface properties of Bacillus licheniformis according to age, temperature and aeration; Herben PF et al.; The surface properties of Bacillus licheniformis DSM 603 have been examined as a function of age during cultures in baffled and non-baffled Erlenmeyer flasks, at 30 and 37 degrees C . The elemental composition of the surface was determined by X-ray photoelectron spectroscopy and its variation was compared with the electrophoretic mobility and water contact angle . The variations of the surface phosphate concentration during the cultures and according to growth conditions indicate that the determining factor is the total cell concentration . The P/C atomic concentration ratio remains in a range where its variation does not influence the electrophoretic mobility . The decrease and increase of hydrophobicity during cultures at 30 degrees C in baffled and normal flasks, respectively, are paralleled by a variation in the O/C ratio.

Science, 1990 Feb 23, 247(4945), 946 - 8
The MerR metalloregulatory protein binds mercuric ion as a tricoordinate, metal-bridged dimer; Helmann JD et al.; Bacterial MerR proteins are dimeric DNA-binding proteins that mediate the Hg(II)-dependent induction of mercury resistance operons . Site-directed mutagenesis of the Bacillus sp . RC607 MerR protein reveals that three of four Cys residues per monomer are required for Hg(II) binding at the single high-affinity binding site . Inactive mutant homodimers can exchange subunits to form heterodimers active for Hg(II) binding . Studies of a heterodimer retaining only three of eight cysteine residues per dimer reveal that Cys79 in one subunit and Cys114 and Cys123 in the second subunit are necessary and sufficient for high-affinity Hg(II) binding in an asymmetric, subunit bridging coordination complex.

J Chromatogr, 1990 Feb 23, 525(2), 307 - 18
Purification of phospholipase-C from Bacillus cereus by affinity chromatography on 2-(4-aminophenylsulphonyl)ethyl-cellulose; Kamberov E et al.; A new method for the purification of phospholipase-C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) from Bacillus cereus has been developed, based on its affinity to 2-(4-aminophenylsulphonyl)ethyl derivative of beaded cellulose . The enzyme was adsorbed on the affinity sorbent through a site(s) that was clearly distinct from its catalytically active site, because it was still active in the immobilized state . A possible role of enzyme-inhibitor interaction in enzyme binding to the ligand used is discussed.

Biochim Biophys Acta, 1990 Feb 23, 1042(3), 410 - 2
Inhibition of phosphatidylinositol-specific phospholipase C by phosphonate substrate analogues; Shashidhar MS et al.; Non-hydrolysable analogues of phosphatidylinositol were synthesized and tested as inhibitors of phosphatidylinositol-specific phospholipase C from Bacillus cereus . In these molecules, the phosphodiester bond of phosphatidylinositol hydrolyzed by the phospholipase was replaced by a phosphonate linkage and a simpler hydrophobic group replaced the diacylglycerol moiety . One of the phosphonates also contained a carboxylate functional group suitable for matrix attachment . All three synthetic phosphonates inhibited the phospholipase C activity in a concentration-dependent manner, with the analogue most closely resembling the structure of the natural substrate, phosphatidylinositol, being the most potent inhibitor . The data indicate that phosphonate analogues of phosphatidylinositol may be useful for study of phospholipase C and other proteins interacting with myo-inositol phospholipids.

Am J Ophthalmol, 1990 Feb 15, 109(2), 127 - 31
Ocular infections associated with Eikenella corrodens; Klein B et al.; Eikenella corrodens is a gram-negative, facultative rod-shaped anaerobe that colonizes the human mouth, nasopharynx, gut, and genitourinary tract . We treated a corneal ulcer from which E . corrodens was the primary isolate in an otherwise healthy man . We treated another patient who had recurrent bacterial endophthalmitis from which Eikenella was identified in mixed culture . Named for its ability to form pits in agar, the corroding bacillus is gaining recognition for its role in head and neck infections . Certain E . corrodens strains are mobile on moist surfaces and elaborate an endotoxin, which may destroy human tissues directly and indirectly by means of the immune system . The organism is usually resistant to aminoglycosides and penicillinase-resistant penicillins yet is susceptible to penicillin and some cephalosporins.

Biochemistry, 1990 Feb 13, 29(6), 1643 - 8
Metal ion dependence of phosphorothioate ATP analogues in the Bacillus stearothermophilus tyrosyl-tRNA synthetase reaction; Garcia GA et al.; Pre-steady-state kinetic analyses on the formation of tyrosyl adenylate from tyrosine and each of the four diastereomers of alpha- and beta-phosphorothioate adenosine triphosphates {ATP alpha S and ATP beta S; Eckstein, F., & Goody, R . (1976) Biochemistry 15, 1685-1691; Yee, D., Armstrong, V . W., & Eckstein, F . (1979) Biochemistry 18, 4116-4123} were performed in the presence of Mg2+, Co2+, and Cd2+ as the divalent metal ion cofactor . A modest preference of 5.5-fold in kappa 3/KA' (where kappa 3 is the rate constant for tyrosyl adenylate formation and KA' is the dissociation constant for ATP, or phosphorothioate ATP, from the E.Tyr.metal.ATP complex) for the Sp ATP alpha S diastereomer and the absence of an inversion of preference when the metal ion is changed suggest that there is a stereospecific enzyme-alpha-phosphate interaction and that there is no direct metal ion interaction with the alpha-phosphate . The extent of reaction of the ATP alpha S diastereomers (30-50%) implies that these analogues are more susceptible to the hydrolytic site reaction previously reported for this enzyme {Wells, T . N . C., & Fersht, A . R . (1986) Biochemistry 25, 1881-1886} . The strong preference in kappa 3/KA' for the RP ATP beta S diastereomer (16-fold for Mg2+ and 50-fold for Co2+) is indicative of a stereospecific interaction with the pro SP beta oxygen of ATP.(ABSTRACT TRUNCATED AT 250 WORDS)

Urology, 1990 Feb, 35(2), 101 - 8
Multicenter study of superficial bladder cancer treated with intravesical bacillus Calmette-Guérin or adriamycin; Khanna OP et al.; We evaluated 155 patients with superficial bladder cancers (Stages Ta, T1, and TIS) and treated them with either intravesical bacillus Calmette-Guerin (Tice strain) (BCG) or doxorubicin hydrochloride (Adriamycin), in a multicenter nonrandomized study . At present 140 of these patients in treatment Groups I and II are being followed up . With additional follow-up, BCG continued to produce a higher percentage of complete remissions (71%) than doxorubicin (54%) . The percentage of incomplete remission with BCG (7%) was half that with doxorubicin (14%) . Half of the patients whose initial therapy failed had complete remission after additional therapy . However, for patients with recurrence, additional follow-up shows a recurrence rate per 100 patient-months for BCG (1.0) only slightly lower than that for doxorubicin (1.1) . The percentage of progressions continued to be higher with BCG (8.5%) than with doxorubicin (5%), but the difference between these results for the two drugs proved slightly less than we reported previously . Of the patients in this study, 2.5 percent (all treated with BCG) required cystectomy . A comparison of the results of our study with those of 13 other studies using BCG to treat bladder cancer indicates that therapy beyond an initial course of 6 weekly treatments increases the percentage of complete response . All of the studies showed that the greatest improvement in percentage of complete response occurred with the second course of treatment . The value of maintenance therapy cannot yet be determined, since few studies have used that protocol . The percentage of patients requiring cystectomy in studies with fewer than 20 treatments was 2.2 times higher than in studies with more than 20 treatments.

Pneumologie, 1990 Feb, 44 Suppl 1, 641 - 3
{Status, development and trends in the epidemiology of tuberculosis in East Germany since 1982}; Schilling W et al.; The article is an epidemiologic review of the situation in the German Democratic Republic with regard to tuberculosis . Conclusions for the future strategy to fight tuberculosis are drawn . During the past few years the drop in the incidence of tuberculosis, especially of bacillary pulmonary tuberculosis, is no longer as marked as it had been previously . Today, tuberculosis is a disease of middle and advanced age, under present prevailing conditions . The strategy for the discovery of new cases of tuberculosis must be adapted to these facts . For surveillance of tuberculosis during the coming years it will be necessary to explore new methods of case finding and of the discovery of sources of infection, screening of high-risk groups and examination of patients with bronchopulmonary signs and symptoms.

J Protein Chem, 1990 Feb, 9(1), 87 - 94
Secondary structure of the entomocidal toxin from Bacillus thuringiensis subsp . kurstaki HD-73; Choma CT et al.; The secondary structure of the toxin from Bacillus thuringiensis subsp . kurstaki (Btk) HD-73 was estimated by Raman, infrared, and circular dichroism spectroscopy, and by predictive methods . Circular dichroism and infrared spectroscopy gave an estimate of 33-40% alpha-helix, whereas Raman and predictive methods gave approximately 20% . Raman and circular dichroism spectra, as well as predictive methods, indicated that the toxin contains 32-40% beta-sheet structure, whereas infrared spectroscopy gave a slightly lower estimate . Thus, all of these approaches are in agreement that the native conformation of Btk HD-73 toxin is highly folded and contains considerable amounts of both alpha-helical and beta-sheet structures . No significant differences were detected in the secondary structure of the toxin either in solution or as a hydrated pellet.

Appl Environ Microbiol, 1990 Feb, 56(2), 340 - 4
Transfer of the toxin protein genes of Bacillus sphaericus into Bacillus thuringiensis subsp . israelensis and their expression; Bourgouin C et al.; The genes encoding the toxic determinants of Bacillus sphaericus have been expressed in a nontoxic and a toxic strain of Bacillus thuringiensis subsp . israelensis . In both cases, the B . sphaericus toxin proteins were produced at a high level during sporulation of B . thuringiensis and accumulated as crystalline structures . B . thuringiensis transformants expressing B . sphaericus and B . thuringiensis subsp . israelensis toxins did not show a significant enhancement of toxicity against Aedes aegypti, Anopheles stephensi, and Culex pipiens larvae.

Am J Epidemiol, 1990 Feb, 131(2), 340 - 8
The effectiveness of bacillus Calmette-Guérin (BCG) vaccination against tuberculosis . A case-control study in Treaty Indians, Alberta, Canada; Houston S et al.; Bacillus Calmette-Guerin (BCG) vaccination against tuberculosis has been used around the world for 60 years, yet its efficacy in large, controlled prospective studies is inconsistent . The factors influencing BCG protection include variation in immunogenic potential, background exposure to environmental mycobacteria, and differences in host response to vaccine . As a means of addressing regional differences in protection, case-control studies provide a relatively inexpensive, rapid means of assessing regional vaccine effects . Treaty Indian cases (n = 160) resident in Alberta, Canada, presenting during a 5-year period (1975-1979) were individually matched for age, sex, and Band with two nontuberculous controls . A 57 percent protection by BCG vaccination was demonstrated . These results support the usefulness of case-control studies and their importance in planning tuberculosis control programs.

Clin Orthop, 1990 Feb, (251), 246 - 8
Fusobacterium osteomyelitis associated with intraosseous gas; Foulkes GD et al.; The diagnosis of acute anaerobic osteomyelitis was made in a 57-year-old hypertensive diabetic woman complaining of groin pain and fever . Roentgenograms and computed tomography demonstrated intraosseous gas in the right femoral head and surrounding soft tissue . Cultures obtained from open biopsy were positive for the anaerobic gram-negative bacillus Fusobacterium necrophorum, a normal inhabitant of the mouth, bowel, and urogenital tract . The patient responded to an antibiotic regimen of metronidazole combined with initial debridement and drainage, followed by resection of the femoral head (Girdle-stone arthroplasty) . The hospital course was complicated by fungal and pseudomonal superinfection . The patient was afebrile and ambulatory at discharge two months after admission . A case of Fusobacterium necrophorum osteomyelitis causing intraosseous gas seems not to have been previously reported in the literature.

Proc Natl Acad Sci U S A, 1990 Feb, 87(3), 1223 - 7
EPR demonstration of iron-nitrosyl complex formation by cytotoxic activated macrophages; Lancaster JR Jr et al.; Activated macrophage cytotoxicity is characterized by loss of intracellular iron and inhibition of certain enzymes that have catalytically active nonheme-iron coordinated to sulfur . This phenomenon involves the oxidation of one of the terminal guanidino nitrogen atoms of L-arginine, which results in the production of citrulline and inorganic nitrogen oxides (NO2-, NO3-, and NO) . We report here the results of an electron paramagnetic resonance spectroscopic study performed on cytotoxic activated macrophage (CAM) effector cells, which develop the same pattern of metabolic inhibition as their targets . Examination of activated macrophages from mice infected with Mycobacterium bovis (strain bacillus Calmette-Guerin) that were cultured in medium with lipopolysaccharide and L-arginine showed the presence of an axial signal at g = 2.039, which is similar to previously described iron-nitrosyl complexes formed from the destruction of iron-sulfur centers by nitric oxide (NO) . Inhibition of the L-arginine-dependent pathway by addition of NG-monomethyl-L-arginine (methyl group on a terminal guanidino nitrogen) inhibits the production of nitrite, nitrate, citrulline, and the g = 2.039 signal . Comparison of the hyperfine structure of the signal from cells treated with L-arginine with terminal guanidino nitrogen atoms of natural abundance N14 atoms or labeled with N15 atoms showed that the nitrosyl group in this paramagnetic species arises from one of these two atoms . These results show that loss of iron-containing enzyme function in CAM is a result of the formation of iron-nitrosyl complexes induced by the synthesis of nitric oxide from the oxidation of a terminal guanidino nitrogen atom of L-arginine.

J Biochem (Tokyo), 1990 Feb, 107(2), 180 - 3
Identification of the reactive cysteinyl residue and ATP binding site in Bacillus cereus glutamine synthetase by chemical modification; Nakano Y et al.; Bacillus cereus glutamine synthetase was modified by reaction with a fluorescent SH reagent, N-{{(iodoacetyl)amino}ethyl}-5-naphthylamine-1-sulfonic acid (IAEDANS), or an ATP analog, 5'-p-fluorosulfonylbenzoyladenosine (FSBA) . The locations of the specific binding sites of these reagents were identified . IAEDANS inactivated Mg2(+)-dependent activity and activated Mn2(+)-dependent activity . FSBA inactivated only Mn2(+)-dependent activity . Mg2+ plus Mn2(+)-dependent activity was inactivated by IAEDANS or FSBA . Amino acid sequence analysis of the single AEDANS-labeled proteolytic fragment showed the cysteinyl residue at position 306 to be the site of modification . Cys 306 is one of three cysteines that are unique to Bacillus glutamine synthetase . The result suggested that the cysteine has a role in the active site of the enzyme . We also report that the amino acid residue modified by FSBA was the lysyl residue at position 43.

J Gen Microbiol, 1990 Feb, 136 ( Pt 2), 367 - 76
Numerical classification and identification of Bacillus sphaericus including some strains pathogenic for mosquito larvae; Alexander B et al.; Ninety-one strains of Bacillus sphaericus, including representatives of all the established DNA homology groups, related round-spored and oval-spored species, and six strains pathogenic for mosquito larvae, were examined for 155 characters . Numerical analyses (Jaccard coefficient/average linkage clustering) based on the 88 variable features revealed 14 clusters at the 79% similarity level that contained more than one strain and 17 single member clusters . All insect pathogenic strains were recovered in a single cluster and the classification was in accord with an established classification based on DNA sequence homology . Two frequency matrices for probabilistic identification were constructed and tested . A comprehensive matrix comprising 14 mesophilic, round-spored taxa and 27 tests gave good results for identification of hypothetical median organisms, cluster overlap and identifications of representative strains (based on data generated in the classification study) . Reference strains for the 14 taxa and eight additional insect pathogenic strains were examined for the 27 tests and were correctly identified with high scores using this matrix . A second matrix comprising seven taxa and 13 tests also performed well in the theoretical evaluation and correctly identified the reference strains and insect pathogenic strains.

J Biochem (Tokyo), 1990 Feb, 107(2), 324 - 30
Purification and characterization of alpha-L-fucosidase from Bacillus circulans grown on porcine gastric mucin; Tsuji Y et al.; Bacillus circulans isolated from soil was found to produce two types of alpha-L-fucosidase differing in substrate specificity . One was able to liberate L-fucose from porcine gastric mucin (PGM), but not from artificial substrates, including p-nitrophenyl and methyl alpha-L-fucosides, while the other acted not on PGM but on p-nitrophenyl alpha-L-fucoside . The production of the former enzyme was enhanced about 150 times as much by PGM added to the medium as by glucose . The alpha-L-fucosidase acting on PGM was purified from the culture fluid obtained with PGM medium by ammonium sulfate fractionation and subsequent column chromatography . The purified enzyme was found to be homogeneous by PAGE and its molecular weight was estimated to be approximately 285,000 . The optimum pH was found to be 5.5 to 6.5 and the stable pH range was 4.5 to 9.0 . The enzyme decomposed various blood group O(H) active substances such as PGM, human milk and human saliva, and moreover acted on A-, B-, and O-erythrocytes . The enzyme was shown to cleave alpha-(1----2)-, (1----3)-, and (1----4)-L-fucosidic linkages in various glycoproteins and oligosaccharides, but failed to hydrolyze alpha-(1----6)-L-fucosic linkages in 6-O-alpha-L-fucopyranosyl-N-acetylglucosamine and intact bovine thyroglobulin.

J Biochem (Tokyo), 1990 Feb, 107(2), 267 - 72
Site-directed mutagenesis of a thermostable alpha-amylase from Bacillus stearothermophilus: putative role of three conserved residues; Vihinen M et al.; The relationship between structure, activity, and stability of the thermostable Bacillus stearothermophilus alpha-amylase was studied by site-directed mutagenesis of the three most conserved residues . Mutation of His-238 to Asp involved in Ca2+ and substrate binding reduced the specific activity and thermal stability, but did not affect the pH and temperature optima . Replacement of Asp-331 by Glu in the active site caused almost total inactivation . Interestingly, in prolonged incubation this mutant enzyme showed an altered end-product profile by liberating only maltose and maltotriose . Conservative mutation of the conserved Arg-232 by Lys, for which no function has yet been proposed, resulted in lowered specific activity: around 12% of the parental enzyme . This mutant enzyme had a wider pH range but about the same temperature optimum and thermal stability as the wild-type enzyme . Results obtained with different mutants were interpreted by computer aided molecular modeling.

Gene, 1990 Jan 31, 86(1), 113 - 7
Structure of the gene encoding beta-1,3-glucanase A1 of Bacillus circulans WL-12; Yahata N et al.; The nucleotide sequence of the glcA gene encoding the precursor of extracellular beta-1,3-glucanase (beta Gl) A1, a polysaccharidase produced by Bacillus circulans WL-12, was determined . The putative glcA gene was 2046 bp long, encoding a polypeptide of 682 amino acids (aa) . The N-terminal aa sequence of beta Gl produced in Escherichia coli harboring the glcA plasmid was identical to that of beta Gl A1 prepared from the culture fluid of B . circulans WL-12 . In both proteins, cleavage of the signal sequence of pre-beta Gl occurred between Ala-38 and Ala-39 of the predicted sequences.

Biochem Biophys Res Commun, 1990 Jan 30, 166(2), 667 - 72
A single amino acid substitution in lactate dehydrogenase improves the catalytic efficiency with an alternative coenzyme; Feeney R et al.; Using site-directed mutagenesis, the NADH-linked lactate dehydrogenase from Bacillus stearothermophilus has been specifically altered at a single residue to shift the coenzyme specificity towards NADPH . The single change is at position 53 in the amino acid sequence where a conserved aspartate has been replaced by a serine . This substitution was made to reduce steric hindrance on binding of the extra phosphate group of NADPH and to remove the negative charge of the aspartate group . The resultant mutant enzyme is 20 times more catalytically efficient than the wild-type enzyme with NADPH.

Biochem Biophys Res Commun, 1990 Jan 30, 166(2), 630 - 6
Nucleotide sequence of the raw-starch-digesting amylase gene from Bacillus sp . B1018 and its strong homology to the cyclodextrin glucanotransferase genes; Itkor P et al.; The gene coding for the raw-starch-digesting amylase from Bacillus sp . B1018 was cloned into Escherichia coli, and its nucleotide sequence was determined . Starting at an ATG codon, there was an open reading frame composed of 2139 bp (713 amino acids) . The NH2-terminal portion encoded a 27 amino acid-long signal peptide . S1 mapping revealed the presence of a typical promoter region upstream from the transcription initiation site . The deduced amino acid sequence of the extracellular mature enzyme was very similar to those of cyclodextrin glucanotransferases (CGTases), especially that of alkalophilic Bacillus sp . 1011 (87% homology) . In fact, the B1018 amylase showed CGTase activity . The COOH-terminal portion of the B1018 amylase shows significant homology with other raw-starch-digesting enzymes.

J Biol Chem, 1990 Jan 25, 265(3), 1369 - 75
The Bacillus thuringiensis delta-endotoxin . Evidence for a two domain structure of the minimal toxic fragment; Convents D et al.; The conformational characteristics of the minimal toxic fragment of the delta-endotoxin from Bacillus thuringiensis berliner 1715 were examined by fluorescence and circular dichroism spectroscopy . This insecticidal protein, specifically toxic to lepidopteran species, was found to consist of two structural domains . Experimental evidence for this conclusion was provided by biphasic guanidine hydrochloride unfolding curves at different pH values and electrophoretic patterns of protease digests . Two stable fragments of comparable molecular weight were obtained using four different broad specificity proteolytic enzymes . A secondary structure model was constructed using seven B . thuringiensis toxin sequences . These toxins were selected on the basis of their limited sequence homology and represent all known insecticidal specificities . Despite this divergence, a consensus secondary structure pattern was obtained, confirming the structural homology among the toxins . The N-terminal halves of all toxins are predicted to be relatively rich in alpha-helix structure and the C-terminal parts to contain alternating beta-strand and coil structures . The latter seems characteristic for a beta-sheet conformation . Comparing this model to the unfolding data obtained by circular dichroism, whose far UV signal gives a measure of the alpha-helix content, allowed us to delineate the structural domains into the primary structure.

Biochim Biophys Acta, 1990 Jan 16, 1042(1), 107 - 12
Bioconversion of leukotriene D4 by lung dipeptidase; Campbell BJ et al.; Sheep lung dipeptidase was released from a lung membrane preparation by digestion with phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis . The total enzyme activity released into the supernatant was 4- to 5-fold greater than that measured in the intact membrane prior to solubilization . The release of the peptidase from the membrane by this treatment is typical of proteins anchored to the lipid bilayer by a covalent attachment of phosphatidylinositol via a C-terminal glycolipid extension . The solubilized lung peptidase was further purified by ammonium sulfate fractionation followed by affinity chromatography and high-pressure liquid chromatography . A linear relationship between log molecular weight and elution volume for proteins of known molecular weight was established using a Toya Soda TSK 3000 high-pressure liquid chromatography column, and the molecular weight of the lung dipeptidase was estimated at 105,000 . The peptidase activity against glycyldehydrophenylalanine of the purified enzyme co-chromatographed in high-pressure liquid chromatography with the activity that converted leukotriene D4 to leukotriene E4 . In kinetic studies using leukotriene D4 as substrate, the relationship between the rate of hydrolysis and enzyme concentration was shown to be linear over the range 20 ng to 98 ng enzyme . Values of Km and Vmax for the dipeptidase using leukotriene D4 as substrate were 43 +/- 6 microM and 11,200 +/- 400 nmol/min per mg, respectively . Inhibition of the conversion of leukotriene D4 to leukotriene E4 was observed with a series of inhibitory agents . Cilastatin, bestatin and chloracetyldehydrophenylalanine were all effective at the micromolar level with cilastatin proving to be the most effective inhibitor . Dithiothreitol was effective within the millimolar range.

Biochem Biophys Res Commun, 1990 Jan 15, 166(1), 61 - 5
Conserved residues of liquefying alpha-amylases are concentrated in the vicinity of active site; Vihinen M et al.; Three dimensional structure of three liquefying type Bacillus alpha-amylases were modeled based on sequence analyses and refined structure of Aspergillus oryzae enzyme . The models suggest that the overall folding motif of alpha-amylases is conserved . The active site, substrate binding and stabilizing calcium binding residues are conserved and concentrated in a cleft between two domains . They constitute the core of alpha-amylases to which other, less conserved regions are attached . The bacterial enzymes have a loop of about 45 residues near the active site and Ca2+ binding region . The loop may be important for the liquefying function of these enzymes.

Mol Biochem Parasitol, 1990 Jan 15, 38(2), 199 - 209
Identification and characterization of glycosylphosphatidylinositol-linked Schistosoma mansoni adult worm immunogens; Sauma SY et al.; Metabolic radiolabeling of adult worms of Schistosoma mansoni with {3H}myristic acid has revealed that the fatty acid is incorporated into more than 15 proteins . We have shown that two of these proteins, a 200-kDa glycoprotein known to be exposed on the surface of the adult worm following praziquantel treatment and a 22-kDa glycoprotein that shows an enhanced immune reactivity with sera of vaccinated mice, are anchored to the adult worm membrane via a glycosylphosphatidylinositol (GPI) linkage . Both antigens partitioned preferentially into the detergent phase of Triton X-114 and were susceptible, following immunoaffinity purification, to hydrolysis by phosphatidylinositol-specific phospholipase C (PIPLC) from Bacillus thuringiensis and phospholipase C from Bacillus cereus . Diacylglycerol (DAG) was released following hydrolysis by bacterial PIPLC; however, Trypanosoma brucei GPIPLC failed to release the diacylglycerol from either protein . Treatment with nitrous acid generated phosphatidylinositol (PI) from both proteins, and phospholipase D from rat serum cleaved phosphatidic acid from the 200-kDa protein . Although the functional significance of these GPI-anchored proteins is unknown, their release from the surface of the schistosome may contribute to immune evasion.

J Immunol, 1990 Jan 15, 144(2), 593 - 8
Phorbol esters increase synthesis of decay-accelerating factor, a phosphatidylinositol-anchored surface protein, in human endothelial cells; Bryant RW et al.; A number of cell-surface proteins are anchored by a phosphatidylinositol (PI)-glycan moiety . These proteins can be released by PI-specific phospholipases C (PI-PLC) . Decay-accelerating factor (DAF) is such a cell-surface protein that protects cells from inadvertent complement attack by binding to and inactivating C3 and C5 convertases . We have studied the regulation of DAF synthesis in human umbilical vein endothelial cells (HUVEC), a cell that has the highest level of surface DAF among those human cells that have been studied . HUVEC DAF was measured by immunoradiometric assay of detergent extracts and of cell supernatants after treatment of cells with a bacterial (Bacillus thuringiensis) PI-PLC . Eighty percent of the HUVEC DAF (4 to 8 x 10(5) molecules/cell) was released by exogenously added PI-PLC, indicating that it is predominantly PI-anchored . The level of PI-PLC-sensitive HUVEC DAF was increased three- to fourfold by overnight treatment of cultures with the protein kinase C activators, PMA (1 to 10 nM), phorbol-12,13-dibutyrate (10 to 100 nM), and teleocidin A (1 to 10 nM) under conditions where cell number, protein, and lactate dehydrogenase remain unchanged . This DAF synthesis was blocked by the protein kinase C inhibitor K-252a in a dose-dependent manner (ED50 = 0.06 microM) . The biologically inactive phorbols, 4-alpha-phorbol-12 myristate-13-acetate (1 microM) and 4-alpha-phorbol-12, 13-didecanoate (1 microM) did not increase DAF levels . The newly expressed DAF in PMA-stimulated cells was still largely PI-anchored . In contrast, another PI-anchored protein, alkaline phosphatase, was not altered by PMA treatment, demonstrating that the PMA effect is not uniform among all surface proteins . The increased expression of DAF only was evident 8 h after PMA addition and was blocked by the RNA and protein synthesis inhibitors, actinomycin D and cycloheximide, indicating that both transcription and translation are required for DAF synthesis induced by phorbol esters . It is concluded that protein kinase C activators cause selective induction of endothelial cell DAF and that DAF synthesis involves protein kinase C activation.

Eur J Biochem, 1990 Jan 12, 187(1), 31 - 8
Molecular forms of acetylcholinesterase in two sublines of human erythroleukemia K562 cells . Sensitivity or resistance to phosphatidylinositol-specific phospholipase C and biosynthesis; Toutant JP et al.; Acetylcholinesterase (AChE) in K562 cells exists in two molecular forms . The major form, an amphiphilic dimer (G2a) which sediments at 5.3 S, and the minor form, an amphiphilic monomer (G1a) which sediments at 3.5 S . Extraction in the presence of the sulfhydryl alkylating agent N-ethylmaleimide was required to preserve the G2a form . In Triton X-100 extracts of the subline K562-243, phosphatidylinositol-specific phospholipase C (PtdIns-PLC) from Bacillus thuringiensis converted most of the G2a AChE into a hydrophilic dimer (G2h), indicating that the G2a form possessed a hydrophobic glycoinositol phospholipid that mediated its attachment to the membrane . Treatment of intact K562-243 cells with PtdIns-PLC released approximately 60% of the total AChE activity and provided an estimate of the externally exposed AChE . The direct conversion from an amphiphilic to a hydrophilic dimeric form by PtdIns-PLC was not obtained in extracts or intact cells of the subline K562-48 . Instead, pretreatment with alkaline hydroxylamine was necessary to render the amphiphilic G2 form of this subline susceptible to digestion by the phospholipase . In this respect, the amphiphilic dimer of K562-48 AChE resembles the G2a form of human erythrocyte AChE, which is resistant to PtdIns-PLC because of the direct palmitoylation of an inositol hydroxyl group in the anchor {Roberts et al . (1988) J . Biol . Chem . 263, 18766-18775} . Release of this acyl chain by hydroxylamine renders the enzyme susceptible to PtdIns-PLC {Toutant et al . (1989) Eur . J . Biochem . 180, 503-508} . In both K562 sublines, sialidase decreased the migration of the G2a form but not of the G1a form of AChE . G1a forms thus appear to represent an intracellular pool of newly synthesized molecules residing in a compartment proximal to the trans-Golgi apparatus . The sialidase-resistant G1a molecules were also resistant to PtdIns-PLC digestion; possible explanations for this resistance are presented.

Science, 1990 Jan 5, 247(4938), 72 - 4
Mechanism of insect resistance to the microbial insecticide Bacillus thuringiensis; Van Rie J et al.; Receptor binding studies show that resistance of a laboratory-selected Plodia interpunctella strain to a Bacillus thuringiensis insecticidal crystal protein (ICP) is correlated with a 50-fold reduction in affinity of the membrane receptor for this protein . The strain is sensitive to a second type of ICP that apparently recognizes a different receptor . Understanding the mechanism of resistance will provide strategies to prevent or delay resistance and hence prolong the usefulness of B . thuringiensis ICPs as environmentally safe insecticides.

J Clin Invest, 1990 Jan, 85(1), 62 - 7
Fibronectin-mediated Calmette-Guerin bacillus attachment to murine bladder mucosa . Requirement for the expression of an antitumor response; Kavoussi LR et al.; Adjuvant intravesical Calmette-Guerin bacillus (BCG) is an effective treatment for superficial bladder cancer . The mechanisms by which BCG mediates antitumor activity are not known . We investigated the initial interaction of BCG with the bladder mucosa to determine whether binding was essential for the development of antitumor activity . Herein, we show that bladder urothelial disruption induced by acrolein, adriamycin, or electrocautery resulted in BCG binding in areas of urothelial damage . Binding induced by each method was inhibited by anti-fibronectin (FN) antibodies but not by antibodies to the basement membrane component laminin . Intravesical BCG binding also was inhibited by pretreating BCG with soluble FN . Inhibition of intravesical FN-mediated BCG attachment prevented immunization via the intravesical route . Moreover, the expression of both delayed hypersensitivity in the bladder of BCG-immunized mice and antitumor activity was inhibited by blocking FN-mediated intravesical BCG attachment . These data suggest that intralumenal attachment of BCG appears to be mediated by FN . Moreover, these data suggest that intravesical FN mediated attachment of BCG is a requisite step in BCG-mediated antitumor activity in the murine bladder tumor model.

Acta Leiden, 1990, 59(1-2), 95 - 112
Maintenance of effective control of Simulium damnosum in the face of insecticide resistance; Kurtak DC; The control of onchocerciasis through reduction of its vector Simulium damnosum depends on highly effective insecticide treatments directed against the larvae . As these treatments must be applied weekly over wide areas for many years, ideal conditions for the development of resistance have been created . Resistance to organophosphate compounds has occurred . To maintain effective vector control, an intense operational research effort has been necessary . Close and constant monitoring of susceptibility, combined with cytotaxonomic identification of the tested larvae, has revealed the pattern of spread of the resistance by migration and apparently by hybridization between populations . These studies have helped indicate when it might be possible to contain or eliminate a resistant population . Screening of all available appropriate compounds has led to the selection of replacement compounds in the microbial (Bacillus thuringiensis H14), pyrethroid (permethrin), and carbamate (carbosulfan) groups . Laboratory studies have revealed esterase and multi-function oxidase mechanisms for the existing organophosphate resistance . Cross-resistance tests have shown no cross-resistance to carbamate insecticides with these mechanisms, but negative correlations with most pyrethroids . These considerations, along with estimated risk of future resistance, effects on non-target organisms, and costs have led to a complex alternation pattern of insecticides, with Bacillus thuringiensis H14 being used in the dry season and a series of classical chemical compounds in the wet season . Through these measures, vector control remains a viable method of onchocerciasis control for the WHO Onchocerciasis Control Programme in West Africa.

Cancer Immunol Immunother, 1990, 31(5), 297 - 304
Major-histocompatibility-complex-class-II-positive cells and interleukin-2-dependent proliferation of immune T cells are required to reject carcinoma cells in the guinea pig; Steerenberg PA et al.; Tumor immunity induced by bacillus Calmette-Guerin was studied in the line 10 hepatocellular carcinoma (line 10) in the strain-2 guinea pig . Line 10 immunity was investigated in vitro with a lymphocyte proliferation assay using line 10 tumor protein extracted with 3 M KCl and in vivo by adoptive transfer of line-10-immune spleen cells . Monoclonal antibodies against guinea pig leucocyte markers were used to block functional properties of the immune cells in order to determine which cell types or cell markers are involved in the immune response to the line 10 tumor . In vitro cells from the spleen, peripheral blood and regional lymph node of immune animals reacted with a proliferative response to line 10 protein . This antigen-specific response was caused by T cells and was regulated by major histocompatibility complex (MHC) class II molecules . In blocking experiments it was found that CT5 (anti-PanT), or MSgp4 {anti-(MHC class I antigen)} monoclonal antibodies did not block but sometimes stimulated the proliferative response . The effect of H159 (anti-PanT) was irregular, while H155 {anti-(T helper)}, and 5C3 {anti-(IL-2 receptor)} monoclonal antibodies blocked the response almost completely . We studied the relevance of the results in vitro obtained and found that mAb 5C3 {anti-(IL-2 receptor)} inhibited the adoptive transfer of line 10 immunity, suggesting that the rejection of line 10 cells is caused by a mechanism that is interleukin-2 (IL-2)-dependent . Moreover, complement lysis of MHC-class-II-antigen-positive immune spleen cells inhibited completely the rejection of the line 10 tumor cell challenge in the adoptive-transfer experiments . In conclusion, our data show that MHC class II molecules or cells possessing these molecules are involved in immunity against line 10 tumor cells, as (a) monoclonal antibodies against MHC class II antigens inhibited the in vitro proliferative response of T cells to tumor antigens and (b) removal of MHC-class-II-positive immune spleen cells abrogated the antitumor effect in the adoptive-transfer experiments . Interleukin-2-dependent proliferation of immune T cells is required for the rejection of line 10 tumor cells.

Nahrung, 1990, 34(2), 177 - 80
Heat resistance of Bacillus spores exposed to food processing conditions; Pendurkar SH et al.; The heat resistance of the spores of 5 Bacillus species was studied in distilled water and pasteurized skim milk . The spores were also subjected to frying and cooking conditions used in fried rice preparations . In presence of milk, the heat resistance of all the five Bacillus species was found to decrease compared to distilled water . Spores of all five Bacillus survived cooking conditions of rice . Frying and subsequent cooking conditions inactivated the spores of all five Bacillus species.

Artery, 1990, 17(4), 189 - 201
Warfarin antagonism of natto and increase in serum vitamin K by intake of natto; Kudo T; Significantly elevated Thrombo-test values after natto intake were observed in patients undergoing anticoagulant therapy after valve replacement operations . Corroborative tests with humans and rabbits to check the observations showed natto to be strongly antagonistic to warfarin . The author suspected that Bacillus natto contained in natto might produce a large amount of vitamin K in the intestinal tracts after taken in, and assumed that the antagonism of natto to Warfarin was attributed to the synthesis of vitamin K by the bacterium in the organ . The author then determined vitamin K in plasma of healthy persons after intake of natto to indicate increases of serum vitamin K . In anticoagulant therapy it is important to maintain the TT value within a certain therapeutic range for a long period of time . Thus the author believes that patients under administration of Warfarin should be given special instructions in food.

Nauchnye Doki Vyss Shkoly Biol Nauki, 1990, (2), 119 - 26
{The quantitative determination of Bacillus thuringiensis beta-exotoxin in insecticidal biopreparations}; Efimtsev EI et al.; A method of quantitative determination of beta-exotoxin content in liquid and dry bioformulations has been developed . The method includes a thin-layer chromatography to isolate beta-exotoxin from accompanying nucleotides, the further desorption of a single beta-exotoxin spot by water and to carry out spectrophotometry at 259 and 330 nm . beta-exotoxin content in industrial formulations bitoxibacillin and turingin I has been determined . The results obtained correspond to the NMR 1H spectroscopy data within the experimental errors . The relative error is 1-2% . The method sensitivity of 0.05 mg/ml . beta-exotoxin content at biotechnological stages of bitoxibacillin production has been determined.

Parasitol Res, 1990, 76(5), 435 - 9
Effect of oltipraz on the susceptibility of adult Schistosoma mansoni to killing by mouse peritoneal exudate cells; Mkoji GM et al.; Incubation of the adult Schistosoma mansoni with the anti-schistosomal compound oltipraz (OPZ) (40 nM) resulted in a significant decrease in schistosome-reduced glutathione (GSH), a thiol compound which may have a role in protection against oxidant-mediated damage . A significant proportion (20-47%) of worms treated with OPZ became susceptible to in vitro killing by zymosan-stimulated peritoneal exudate cells from mice infected with S . mansoni or inoculated with Bacillus Calmette Guerin (BCG) . Killing of the worms was partially inhibited by the addition to the assay system of exogenous glutathione peroxidase with GSH but not by superoxide dismutase . These results suggested that killing of parasites exposed to the drug was partly mediated by cell-generated hydrogen peroxide . They indicate also that depletion of schistosome GSH levels could render the parasites susceptible to killing by oxidative mechanisms, and suggest that there is potential in exploiting schistosome oxidant defense systems and reactive oxygen byproducts in the treatment of schistosomiasis . Inhibition of schistosome oxidant defense systems with drugs may render the parasites susceptible to killing by reactive oxygen byproducts of effector cells.

Mikrobiol Zh, 1990 Jan-Feb, 52(1), 10 - 4
{Hydrolase biosynthesis by morphological variants of Bacillus mesentericus as dependent on the storage conditions}; Koltukova NV et al.; A method used to prepare the inoculum (a strain of Bacillus mesentericus, a producer of a complex of hydrolytic enzymes) has been studied for its effect on the activity of proteinases and amylases under submerged cultivation in fermenters . Optimal conditions for the culture storage and inoculum cultivation are developed to obtain standard enzymic preparations.

Mol Biol (Mosk), 1990 Jan-Feb, 24(1), 244 - 7
{BsiI--a new unusual restriction endonuclease}; Degtiarev SKh et al.; The restriction endonuclease BsiI from Bacillus sphaericus was isolated . The recognition sequence and cleavage point of enzyme BsiI have been determined as (sequence: see text) . This restriction endonuclease is not an isoschizomer of any known restriction endonucleases and differs from other enzymes: it hydrolyses DNA into unsymmetrical recognition sequence.

Proteins, 1990, 7(1), 74 - 92
Structure determination and refinement of Bacillus stearothermophilus lactate dehydrogenase; Piontek K et al.; Structures have been determined of Bacillus stearothermophilus "apo" and holo lactate dehydrogenase . The holo-enzyme had been co-crystallized with the activator fructose 1,6-bisphosphate . The "apo" lactate dehydrogenase structure was solved by use of the known apo-M4 dogfish lactate dehydrogenase molecule as a starting model . Phases were refined and extended from 4 A to 3 A resolution by means of the noncrystallographic molecular 222 symmetry . The R-factor was reduced to 28.7%, using 2.8 A resolution data, in a restrained least-squares refinement in which the molecular symmetry was imposed as a constraint . A low occupancy of coenzyme was found in each of the four subunits of the "apo"-enzyme . Further refinement proceeded with the isomorphous holo-enzyme from Bacillus stearothermophilus . After removing the noncrystallographic constraints, the R-factor dropped from 30.3% to a final value of 26.0% with a 0.019 A and 1.7 degrees r.m.s . deviation from idealized bond lengths and angles, respectively . Two sulfate ions per subunit were included in the final model of the "apo"-form--one at the substrate binding site and one close to the molecular P-axis near the location of the fructose 1,6-bisphosphate activator . The final model of the holo-enzyme incorporated two sulfate ions per subunit, one at the substrate binding site and another close to the R-axis . One nicotinamide adenine dinucleotide coenzyme molecule per subunit and two fructose 1,6-bisphosphate molecules per tetramer were also included . The phosphate positions of fructose 1,6-bisphosphate are close to the sulfate ion near the P-axis in the "apo" model . This structure represents the first reported refined model of an allosteric activated lactate dehydrogenase . The structure of the activated holo-enzyme showed far greater similarity to the ternary complex of dogfish M4 lactate dehydrogenase with nicotinamide adenine dinucleotide and oxamate than to apo-M4 dogfish lactate dehydrogenase . The conformations of nicotinamide adenine dinucleotide and fructose 1,6-bisphosphate were also analyzed.

Protein Eng, 1990 Jan, 3(3), 181 - 91
Random mutagenesis used to probe the structure and function of Bacillus stearothermophilus alpha-amylase; Holm L et al.; Mutations that cover the sequence of Bacillus stearothermophilus alpha-amylase were produced by an efficient in vitro enzymatic random mutagenesis method and the mutant alpha-amylases were expressed in Escherichia coli, which also secreted the product . Ninety-eight mutants were identified by sequencing and their enzyme activities were classified into three classes: wild-type, reduced or null . A molecular model of the enzyme was constructed using the coordinates of Takaamylase A and a consensus alignment of mammalian, plant, and bacterial alpha-amylases . The location of mutant amino acids on the model indicate that mutations which destroy or decrease the catalytic activity are particularly clustered: (i) around the active site and along the substrate-binding groove and (ii) in the interface between the central alpha/beta barrel and the C-terminal domain . Exposed loops are typically tolerant towards mutations.

Proteins, 1990, 7(2), 156 - 71
Beta-lactamase of Bacillus licheniformis 749/C at 2 A resolution; Moews PC et al.; Two crystal forms (A and B) of the 29,500 Da Class A beta-lactamase (penicillinase) from Bacillus licheniformis 749/C have been examined crystallographically . The structure of B-form crystals has been solved to 2 A resolution, the starting model for which was a 3.5 A structure obtained from A-form crystals . The beta-lactamase has an alpha + beta structure with 11 helices and 5 beta-strands seen also in a penicillin target DD-peptidase of Streptomyces R61 . Atomic parameters of the two molecules in the asymmetric unit were refined by simulated annealing at 2.0 A resolution . The R factor is 0.208 for the 27,330 data greater than 3 sigma (F), with water molecules excluded from the model . The catalytic Ser-70 is at the N-terminus of a helix and is within hydrogen bonding distance of conserved Lys-73 . Also interacting with the Lys-73 are Asn-132 and the conserved Glu-166, which is on a potentially flexible helix-containing loop . The structure suggests the binding of beta-lactam substrates is facilitated by interactions with Lys-234, Thr-235, and Ala-237 in a conserved beta-strand peptide, which is antiparallel to the beta-lactam's acylamido linkage; an exposed cavity near Asn-170 exists for acylamido substituents . The reactive double bond of clavulanate-type inhibitors may interact with Arg-244 on the fourth beta-strand . A very similar binding site architecture is seen in the DD-peptidase.

Jordemodern, 1990 Jan-Feb, 103(1-2), 24 - 7
{Scholarship report . Midwife in Chile}; Bonilla E; PIP: Midwifery training was started in Chile in 1834 . In 1877 a pediatric institute was founded, and a professional organization for midwives was formed in 1919 . In 1958 midwives were charged with BCG (bacillus Calmette-Guerin) vaccination of infants . A new professional organization was formed in 1962 . In 1965 family planning instruction was initiated in maternity wards because of the high mortality rate after illegal abortions . 5 years later programs were launched on cervical, uterine, and breast cancer for midwives . In 1974 midwives became responsible for contraceptive counseling in a campaign to prevent sexually transmitted diseases . After 1973 the new government cut public health care outlays and staff was reduced by attrition . In 1984 the unemployment rate among newly graduated midwives stood at 34% . After privatization of hospitals 20% of the population got private health care paid by insurance . The employment of midwives increased but mostly in administrative and subservient roles assisting doctors in deliveries . A normal delivery costs $893 and a cesarean $1250, while the net earnings of a midwife amount to $176 a month . Caesareans make up 60-70% of all deliveries, and they are a major cause of complications associated with anesthesia . The state-run El Salvador hospital in Santiago has 11,000 deliveries a year . There had not been a maternal death from childbirth in the previous 19 months . Neonatal mortality is 5.3/1000 live births, and 12% of pregnant women have high blood pressure and 9% have hepatosis . The average stay is 2.5 days . The rate of caesareans (24.5%) is expected to diminish to 15% . Midwives work 12 hours at a stretch under adverse conditions and stress . A new proposal submitted to the University of Chile wants to abolish midwifery training altogether and replace it by some courses in obstetrics, gynecology, and pediatrics in the nursing curriculum .

FEMS Microbiol Lett, 1990 Jan 1, 54(1-3), 95 - 9
Heterologous expression of a mutated toxin gene from Bacillus thuringiensis subsp . tenebrionis; Rhim SL et al.; Using oligonucleotide probes we have isolated a DNA fragment encoding an insecticidal toxin of the coleopteran specific Bacillus thuringiensis subsp . tenebrionis . The gene was altered by site directed mutagenesis at its 5'-end and adapted for general cloning and expression purposes with a linker including a start codon and new restriction sites . The constructs were inserted into several vector plasmids and expressed in Escherichia coli . Expression E . coli was strongly enhanced by the lac-promoter . A fusion protein with phage MS2-polymerase was produced together with a 67 kDa protein also found for normal expression of the toxin gene . Synthesis of the latter protein indicated a second ribosome binding site at the 5'-terminus of the toxin encoding sequence . Toxin-containing proteins were identified by Western blot analysis . The positive cell extracts from E . coli had insecticidal activity on larvae of the Colorado potato beetle . The cloned gene is not homologous to a gene previously cloned by us whose gene products were also toxic to coleopteran larvae.

Z Versuchstierkd, 1990, 33(1), 29 - 35
Seromonitoring in small laboratory animal colonies . A five year survey: 1984-1988; Kraft V et al.; From 1984 to 1988 one thousand serologic investigations of laboratory animal colonies originating from 10 different European countries were performed . The most prevalent infections in mouse stocks were caused by Mouse hepatitis virus (MHV), Minute virus of mice (MVM), Theiler's encephalomyelitis virus (TEMV), Reovirus type 3 (Reo3), Sendaivirus, and Pneumonia virus of mice (PVM) . In mice no infections with Lymphocytic choriomeningitis virus (LCM), Polyomavirus, Mouse adenovirus, and K-virus were recorded . Only two colonies were infected by Ectromelia virus . The first six virus infections of mice were also found in rat colonies as well as the rat-specific Coronaviruses (Sialodacryoadenitisvirus--SDA, Rat corona virus--RCV) and Parvovirus (Kilham rat virus--KRV, Toolan H-1 virus) being endemic . Antibodies to Bacillus piliformis were detectable in about 50% of rat stocks screened . This is in contrast to the mouse, where only about 10% of the colonies were found to be positive . A similar picture was seen for M . pulmonis which is primarily an infection of the rat . In mice no case was detected during the last two years . The number of investigations performed from guineapig, hamster and rabbit colonies was relatively low . Nevertheless, antibodies against the following antigens were detectable: In guineapig stocks: Reo3, PVM, Sendai, Simian virus 5 (SV5) and B . piliformis; in rabbits: Reo3, Sendavirus, SV5, and B . piliformis; in hamsters: PVM, LCM and B . piliformis . The overall contamination rate showed a continuous decrease until 1988 . Nevertheless, about 50% of mouse and rat stocks still exhibited antibodies to one or more viral infections.

Eur Urol, 1990, 17(2), 125 - 8
Bacillus Calmette-Guérin treatment and vesicorenal reflux; Bohle A et al.; 14 patients with radiologically documented vesicorenal reflux (5 bilateral, 7 unilateral, 2 unilateral double-J stents) received intravesical immunotherapy with bacillus Calmette-Guerin (BCG) after treatment of their superficial urothelial tumors . 1 patient with a double-J stent suffered a highly febrile attack following obstruction of the indwelling ureteral catheter . The other patients tolerated 1 or 2 (6 of 14 patients) courses of BCG without an increase in side effects . Topical intravesical therapy with BCG in patients with vesicorenal reflux seems to be indicated in view of the multicentricity of urothelial carcinoma and can be performed without an increased risk of complications.

Acta Crystallogr A, 1990 Jan 1, 46 ( Pt 1), 57 - 68
Direct phase determination for the molecular envelope of tryptophanyl-tRNA synthetase from Bacillus stearothermophilus by X-ray contrast variation; Carter CW Jr et al.; Monoclinic crystals of Bacillus stearothermophilus tryptophanyl-tRNA synthetase grown in the presence of substrate tryptophan (space group P2(1)) display evidence of a low-resolution trigonal space group (P321) . The origin and averaging transformations for the local 32 point group of this unusually clear sixfold non-crystallographic symmetry may be inferred without prior estimation of the electron density . This local symmetry was exploited in conjunction with solvent density contrast variation to determine the shape of the molecular envelope . X-ray intensities measured from crystals equilibrated in mother liquors of three different electron densities were used to estimate three parameters for each reflection: the modulus of the envelope transform, {Gh}; and components, Xh and Yh, relative to Gh, of the structure-factor vector for the transform of intramolecular density fluctuations . The moduli ({Gh}) behave somewhat like structure-factor amplitudes from small-molecule crystals, and estimation of their unknown phases was successfully carried out by statistical direct methods . Reflections to 18 A resolution, which obey rather well the symmetry of space group P321, were merged to produce an asymmetric unit in that space group . {Gh} values for the 34 strongest of these were phased using the small-molecule direct-methods package MITHRIL {Gilmore (1984) . J . Appl Cryst . 17, 42-46} . The best phase set was expanded back to the P2(1) lattice and negative density was truncated to generate initial phases for all reflections to 18 A resolution . Phase refinement by iterative imposition of the local 32 symmetry produced an envelope with convincing features consistent with known properties of the enzyme . The envelope implies that the tryptophanyl-tRNA synthetase dimer is an elongated structure with an axial ratio of about 4:1, in which the monomers have two distinct domains of unequal size . The smaller of these occurs at the dimer interface, and resembles the nucleotide binding portion of the tyrosyl-tRNA synthetase . It may therefore contain the amino-terminal one hundred or so residues, including all three cysteines, previously suggested to comprise a nucleotide-binding domain in the tryptophanyl enzyme . A purely crystallographic test of the overall features of this envelope was carried out by transporting it to a tetragonal crystal form of the same protein in which the asymmetric unit is a monomer . The small domain fits snugly inside three mercury and one gold heavy-atom binding sites for this crystal form; and symmetry-related molecules provide excellent, but very different, lattice contacts in nearly all directions.

Appl Environ Microbiol, 1990 Jan, 56(1), 264 - 8
Model for inactivation and disposal of infectious human immunodeficiency virus and radioactive waste in a BL3 facility; Stinson MC et al.; A method is described for autoclaving low levels of solid infectious, radioactive waste . The method permits steam penetration to inactivate biologic waste, while any volatile radioactive compounds generated during the autoclave process are absorbed . Inactivation of radiolabeled infectious waste has been problematic because the usual sterilization techniques result in unacceptable radiation handling practices . If autoclaved under the usual conditions, there exists a high probability of volatilization or release of radioisotopes from the waste . This results in the radioactive contamination of the autoclave and the laboratory area where steam is released from the autoclave . Our results provide a practical method to inactivate and dispose of infectious radioactive waste . For our research, Bacillus pumilus spore strips and vaccinia virus were used as more heat-resistant surrogates of the human immunodeficiency virus (HIV) . These surrogates were used because HIV is difficult to grow under most conditions and is less heat tolerant than the surrogates . In addition, B . pumilus has defined cell death values, whereas such values have not been established for HIV . Both B . pumilus and vaccinia virus are less hazardous to work with . The autoclave method is time efficient and can be performed by laboratory personnel with minimal handling of the waste . Furthermore, waste site handlers are able to visually inspect the solid waste containers and ascertain that inactivation procedures have been implemented.

Ann Clin Biochem, 1990 Jan, 27 ( Pt 1), 33 - 7
Colorimetric glucose assay using thermostable glucokinase; Scott DA et al.; A method for assaying glucose in serum or plasma samples using a thermostable glucokinase was developed . Glucokinase from Bacillus Stearothermophilus was coupled with glucose-6-phosphate dehydrogenase to produce NADPH, which reduced the tetrazolium dye MTT to its formazan . Detection of the product at 660 nm allowed samples containing up to 30 mmol/L glucose to be assayed with an endpoint method . Use of the optimal wavelength for formazan detection, 570 nm, increased sensitivity for NADPH detection by over threefold compared to UV detection . The stability of glucokinase assay mixtures was extensively studied, with variation in buffers, salt and enzyme stabilizers . Maximal half life for reagent stability at room temperature was approximately 30 days, with storage of assay mixtures in two solutions . Various drugs and metabolites were tested for interference in the method and no significant interferences were found.

Histopathology, 1990 Jan, 16(1), 83 - 8
Epithelioid angiomatosis in the acquired immunodeficiency syndrome: morphology and differential diagnosis; Walford N et al.; A rare vascular proliferation found as a skin lesion in patients suffering from the acquired immunodeficiency syndrome and sometimes referred to as epithelioid angiomatosis is believed to be a manifestation of infection by the cat scratch bacillus or a related organism . We describe the histological findings from eight lesions seen in two cases . In all cases the diagnosis could be confirmed by demonstration within the lesions of groups of gram-negative rod-shaped organisms staining positively with the Warthin-Starry stain . This condition needs to be distinguished from a variety of reactive and neoplastic vascular proliferations.

Am Rev Respir Dis, 1990 Jan, 141(1), 94 - 7
Plasma tumor necrosis factor in patients with septic shock . Mortality rate, incidence of adult respiratory distress syndrome, and effects of methylprednisolone administration; Marks JD et al.; We assayed serial plasma samples from 86 patients, who were enrolled in a prospective randomized trial of the effects of methylprednisolone (MPSS) in septic shock, for the presence of cytokine tumor necrosis factor (TNF) using an enzyme-linked immunosorbent assay . TNF was present in the plasma of 27 of the 74 patients with septic shock, but in only 1 of the 12 patients with shock due to other causes . TNF was detected with equal frequency in patients with shock from gram-negative or from gram-positive bacillary sepsis . TNF levels were highest on the initial sample and decreased significantly over the subsequent 24 h in both the patients treated with MPSS and in those given placebo . Patients with detectable TNF had a higher incidence and severity of the adult respiratory distress syndrome and a higher mortality rate than did patients without detectable TNF.

J Bacteriol, 1990 Jan, 172(1), 511 - 3
Use of both translation initiation sites of the middle wall protein gene in Bacillus brevis 47; Adachi T et al.; The middle wall protein gene of Bacillus brevis 47 has two potential translation initiation sites located tandemly in the same reading frame . We demonstrate here that both sites are utilized to start translation in B . brevis 47 . Translation from the first site (located upstream) gives rise to a precursor of the middle wall protein with an extension peptide of 31 amino acids preceding the signal peptide . The precursor was cleaved at the same position as that of the precursor translated from the second site . The TTG codon seems to play an appreciable role in the initiation of translation in B . brevis 47.

Infect Immun, 1990 Jan, 58(1), 189 - 96
The 75-kilodalton protein of Chlamydia trachomatis: a member of the heat shock protein 70 family?
Danilition SL, Maclean IW, Peeling R, Winston S, Brunham RC.
The gene encoding a 75-kilodalton (kDa) protein of Chlamydia trachomatis was cloned, expressed, and sequenced . Genomic libraries from C . trachomatis serovar D DNA were constructed in vectors pUC18 and lambda gt11 and were screened with a panel of monoclonal antibodies against C . trachomatis antigens . The only recombinants identified were those that reacted with antibody UM-13, which has specificity for a genus-specific epitope on the 75-kDa protein . The gene was localized to a 2.9-kilobase DNA fragment and sequenced . The gene consists of a long open reading frame of 1,956 nucleotides, which translates into 652 amino acids totalling 70,558 daltons in mass . Putative promoter elements and a ribosome binding site were identified within 5'-flanking sequences, and a typical rho-independent terminator was identified within 3'-flanking sequences . Screening of the GenBank nucleic acid sequence data bank revealed extensive similarity between the chlamydial 75-kDa gene and the heat shock protein 70 (hsp70) family or proteins . In particular, 71 and 69% amino acid sequence similarities were identified with hsp70 of Escherichia coli and Bacillus megaterium, respectively . Polyclonal antibodies were produced to the recombinant antigen in rabbits and detected epitopes on elementary bodies in enzyme-linked immunosorbent and indirect microimmunofluorescence assays . Antibodies reacted with an antigen of identical molecular mass in L2 and C serovars in an immunoblot assay and neutralized these serovars in cell culture . The 75-kDa protein appears to be a chlamydial homolog of hsp70, is immunoaccessible on native elementary bodies, and is a target for neutralization.

FEBS Lett, 1990 Jan 1, 259(2), 297 - 300
Identification of a reversible structural transition in the metal-depleted glycerol dehydrogenase from Bacillus stearothermophilus; Spencer P et al.; Evidence is presented to demonstrate that the Zn2+ -depleted, inactive form of the glycerol dehydrogenase from Bacillus stearothermophilus exists in one of two possible conformations in equilibrium, the position of which is temperature sensitive . The conformation of the metal-depleted enzyme favoured by higher temperatures (20-40 degrees C) is able to bind Zn2+ and regain catalytic activity, whereas that favoured at lower temperatures (0-10 degrees C) is unable to bind metal ions and is thus inactive . This equilibrium is also pH dependent with a pK of 6.6 . At pH 6.0, the equilibrium lies in favour of the form of the enzyme able to bind metal ions and exhibit activity.

Comp Immunol Microbiol Infect Dis, 1990, 13(3), 155 - 62
Discrepancy between in vitro and in vivo assays for the susceptibility of Bacillus C.I.P . 5832 to antimicrobial agents; Tirel ML et al.; In vitro, avoparcin, flavomycin, virginiamycin bacitracin, trimethoprim + sulfadimethoxine, colistin, spiramycin, oxolinic acid, lincomycin + spectinomycin and a mixture of organic acids were able to inhibit Bacillus C.I.P . 5832 . In vivo, none of these antibacterial agents, at the levels incorporated in feeds, could affect the viable bacillus count in the caecal content of chickens . In another in vitro assay, olaquindox, carbadox and tylosin were also effective in inhibiting Bacillus C.I.P . 5832 . But, concomitantly administered with carbadox or tylosin, Bacillus C.I.P . 5832 still promoted growth and feed conversion of fattening pigs . These results, as well as data of the quoted literature, suggest that the current in vitro techniques should not be used to screen the susceptibility of Bacillus C.I.P . 5832 to antimicrobial agents . Only in vivo assays seem to be of value.

Probl Tuberk, 1990, (11), 65 - 8
{Indices of humoral immunity in inhabitants of the Far North infected and sick with tuberculosis}; Vasil'ev AV et al.; As a result of a complex study of the humoral immunity indices in tuberculosis infected and affected inhabitants of the Extreme North with reference to the immunoglobulin A, G, M levels, titers of heterohemagglutinins (HHA) and specific antibodies defined by means of indirect hemagglutination test (IHAT), complement fixation test with tuberculin and IHAT with phosphatidic antigen (IHAT-Ph), two simple and quite informative tests, i.e . HHA and IHAT-Ph, are recommended for the practical use in health system . HHA level greater than 1/4 indicates the absence of immunodeficiency in system B . The results of IHAT-Ph, being adequate to the main clinical manifestations of tuberculosis (including bacillary excretion and lung tissue destruction), can be used as an additional diagnostic and evaluation method of tuberculous activity when the subjects from specific infection foci are examined or during their inpatient and dispensary observation and management.

Probl Tuberk, 1990, (11), 21 - 3
{Improvement of chemotherapy of primary forms of pulmonary tuberculosis in adolescents}; Firsova VA et al.; Observation findings concerning 112 adolescents with tuberculosis are presented . The patients were divided into 2 groups . Group 1 included 61 cases with primary forms of tuberculosis and without lung tissue destruction or bacillary excretion . The best part of this group was affected by tuberculosis of intrathoracic lymph nodes and focal tuberculosis . Group 2 consisted of 51 patients with primary tuberculosis in addition to lung tissue destruction or bacillary excretion . Most of these patients had infiltrative tuberculosis . Their treatment regimens were standard ones . The patients of group 1 are recommended to have a 6-month hospital management and a subsequent additional treatment at a sanatorium for 3 months . The period of the in-patient chemotherapy of group 2 should be no less than 9 months plus 3 months of additional treatment at a sanatorium.

Arkh Patol, 1990, 52(7), 48 - 51
{The characteristics of the proliferative processes in the epithelium of the sigmoid intestine in acute bacillary dysentery}; Borovskaia TF et al.; Proliferative processes were studied using radioautography with 3H-thymidine at the height of the disease and in the early convalescence of the bacterial dysentery . The mucous membrane of the sigmoid colon of healthy persons undergoing rectoromanoscopy due to the epidemic reasons served as control . No pathologic changes were found in these healthy persons; the labeled nuclei index (LNI) was 5.75 +/- 0.4%, the label intensity (LI) characterizing the rate of DNA synthesis was 14.9 +/- 0.8 . During the peak of bacterial dysentery LNI increased up to 12.9 +/- 1.3% and there was a three-fold increase of LI . Activation of the proliferative processes persisted in the period of an early recovery . LNI was 9.4 +/- 1.0%, LI was 2.6 times higher as compared to control.

Yi Chuan Xue Bao, 1990, 17(3), 216 - 25
{A novel type of phase variation regarding integrated and free states of plasmid pFDX163 in Bacillus stearothermophilus CU21}; He XS et al.; pFDX1 is a recombinant plasmid which carries a foreign gene xylE . By selecting for kanamycin-resistant mutants of Bacillus stearothermophilus CU21(pFDX1) at higher temperature, a variant strain CU21-163 was obtained . This strain harbors a mutant plasmid pFDX163, which was formed by insertion of a 2.0kb H-fragment from the CU21 genome onto the plasmid pFDX1 . pFDX163 was supposed to be integrated into the CU21 chromosome via homologous recombination of H-fragments . The CU21-163 strain consists of two cell types, i.e . y-cell and w-cell . The expression level of xylE gene in the former is higher than that in the latter . The progeny of a y-cell always contains some w-cells, while that of a w-cell contains y-cells . This is supposed to be due to a phase variation of CU21-163 . Analysis on the amount of free and integrated plasmid DNA in different DNA samples of CU21-163 cells allows us to draw the conclusion that there are both free and integrated plasmids in the y-cells, whereas only integrated ones in the w-cells.

Ann Urol (Paris), 1990, 24(5), 435 - 40
{Results of treatment of superficial vesical tumors by transurethral resection alone and transurethral resection followed by an intravesical instillation of Calmette-Guerin bacillus}; Chopin D et al.; Superficial bladder tumors treated at the Henri Mondor Hospital from 1984 through 1988 were analyzed for recurrence and progression using the following prognostic parameters: stage (TNM classification, 1978), grade (G1, G2, G3), size, number of tumors, and tumor malignancy index as defined by the Besancon group . Forty-five patients were treated with transurethral resection alone (TUR group) whereas 30 had TUR followed by the prophylactic instillation of fresh Calmette-Guerin bacillus in the bladder (BCG group) . In TUR patients, parameters predictive of progression included grade G3, multiple tumors, stage T1, recurrence within 6 months of TUR, and a tumor malignancy index above 455 . None of these criteria were predictive of a response to BCG . Results obtained in the BCG group were comparable to those reported in the literature and confirmed the efficacy of BCG instillations to prevent recurrence and progression of superficial carcinomas of the bladder.

Biochem Int, 1990, 21(4), 741 - 51
Use of a hapten specific anti-dansyl antibody for the localization of ribosomal proteins by immuno electron microscopy; Bergmann U et al.; The fluorescent reagent dansyl chloride has been used as an immunological marker for the electron microscopic localization of ribosomal proteins on the surface of 50S ribosomal subunits . The proteins BstL1 from Bacillus stearothermophilus and EcoL1 from Escherichia coli were dansylated to various degrees and reconstituted into the L1-deficient E . coli 50S subunits from mutant MV17-10 . Using antibodies specific to dansyl chloride, both proteins were mapped at the lateral protuberance near the peptidyl transferase center.

Scand J Urol Nephrol, 1990, 24(3), 191 - 8
Local bacillus Calmette-Guerin therapy for superficial bladder cancer: clinical, histological and ultrastructural patterns; Rigatti P et al.; The effectiveness of intravesical administration of bacillus Calmette-Guerin as a prophylaxis of superficial bladder cancer has been definitely demonstrated . On the other hand, therapeutic regimens, duration effects, efficacy of either maintenance cycles or repeated courses of therapy in case of failures are still controversial . We report the results achieved in 15 cases of carcinoma in situ of the bladder and in 48 cases of superficial bladder cancer (Ta-T1 stage of disease) with bacillus Calmette-Guerin immunotherapy . Our patients underwent an initial six week cycle and a following maintenance cycle with monthly administrations for one year . Median follow-up was 19 months (range 18-21 months) . Patients with carcinoma in situ are now free of disease; on the contrary, patients with Ta-T1 tumors experienced 18 recurrences (28%) . There was a marked decrease of recurrence rate when compared to previous local chemotherapy . We report in detail the adverse effects encountered and both histologic and ultrastructural findings observed after immunotherapy . Bacillus Calmette-Guerin therapy can influence positively the natural history of the disease but possible adverse effects should always be considered before starting the treatment.

Probl Tuberk, 1990, (7), 43 - 5
{Tisamide in the complex treatment of tuberculosis}; Onishchenko VV et al.; The outcomes of tisamid treatment of newly diagnosed patients with bacillary pulmonary tuberculosis were analysed . To estimate the efficacy of tisamid and its administration indications, chemotherapy of the patients was performed with the use of two therapeutic regimens . Isoniazid, rifampicin and streptomycin (or ethambutol) were given to 73 patients, while other 72 ones were treated with the same drugs plus tisamid . The patients in each group were subdivided into slow, rapid and homozygotic (the most rapid) acetylators . Tisamid, when prescribed to newly-discovered patients with destructive tuberculosis, accelerates recovery, by excluding the risk of a hepatotoxic action . Tisamid in a combined treatment with isoniazid, rifampicin and streptomycin is mostly recommended for patients with a rapid acetylation phenotype, i.e . the cases for whom a short-term chemotherapy is possible.

Probl Tuberk, 1990, (7), 28 - 31
{Variants of the course of fibrous-cavernous pulmonary tuberculosis}; Sadykov AS; Taking into account the presence of the process development symptoms or their absence (including a high rate of exacerbations, ongoing dissemination process, formation of new cavities in the affected foci, severe infiltration of the lung tissue and massive bacillary excretion), process development rate and the nature of complications, the following 4 versions of fibrous-cavernous pulmonary tuberculosis were identified: rapidly developing disease; slowly developing disease; fibrous-cavernous pulmonary tuberculosis whose clinical picture was determined by different complications; and relatively stable condition . ++Clinico-roentgenological+ features of each version of the disease are described . Abnormalities in the ++clinico-roentgenological+ picture and laboratory findings are especially apparent in patients with rapidly developing fibrous-cavernous pulmonary tuberculosis.

Bull Pan Am Health Organ, 1990, 24(3), 307 - 13
Field evaluation of Bacillus thuringiensis var . israelensis for control of black flies in the North Littoral Zone of Brazil's São Paulo State; Araujo-Coutinho CJ et al.; The impact of three flowable concentrate formulations of Bacillus thuringiensis var . israelensis (H-14) upon Simulium pertinax larvae was evaluated in 24 coastal streams of Brazil's Sao Paulo State . While no significant differences were found regarding the three formulations' effectiveness, significant correlations were found between the discharge rates of individual streams and the distances over which at least two of the formulations were carried effectively downstream to produce 80% mortality . The relatively short carry distances found for small streams could pose difficulties for control programs that need to treat large numbers of such streams, and suggests a need for research directed at increasing the distances B . thuringiensis (H-14) formulations can be carried.

Int J Syst Bacteriol, 1990 Jan, 40(1), 92 - 7
Taxonomy of alkaliphilic Bacillus strains; Fritze D et al.; The DNA base compositions of 78 alkaliphilic Bacillus strains were determined . These strains were grouped as follows: DNA group A, guanine-plus-cytosine (G+C) content of 34.0 to 37.5 mol% (17 strains); DNA group B, G+C content of 38.2 to 40.8 mol% (33 strains); and DNA group C, G+C content of 42.1 to 43.9 mol% (28 strains) . DNA group A includes the type strain of Bacillus alcalophilus Vedder 1934 . DNA-DNA hybridization studies with DNA group A strains revealed that only one strain, strain DSM 2526, exhibited a high level of DNA homology with B . alcalophilus DSM 485T (T = type strain) . Neither strain DSM 485T nor any other DNA group A strain is homologous to any of the Bacillus type strains with comparable base compositions . Six strains formed a distinct group containing three highly homologous strains and three strains exhibiting greater than 50% DNA homology.

Acta Leprol, 1990, 7(2), 199 - 204
The demystification of leprosy: a multifactorial problem; Kato L; Abolishment of misbeliefs and misconceptions, unfounded fear and prejudice are factors as important in leprosy control as prevention, early detection and therapy . Concrete measures of demystification are proposed . Identify and divulge the absolute truth about leprosy . Calling leprosy "Hansen's disease" did not result in demystification . Patients know that the two terms are identical . Treating them as human beings attracts more patients to the healers than the Hansenologian ritual . Contrary to statements, no major advances are being made in the field of bacteriology, immunology, molecular biology, mode of transmission and epidemiology of leprosy . Not a single new drug has been discovered in 26 years . Vaccination is a dubious venture . The question arises as to whether the right priorities are promoted in leprosy research . Cultivation of the leprosy bacillus is the sine qua non of any further progress . This field of research is a lost and totally neglected priority . Consequently we have no pharmacological model for badly needed of ultrapotent antileprosy drugs . Syphilis is now cured with a single dose of penicillin . A drug as potent against leprosy should not be a mission impossible if an appropriate pharmacological model--the in vitro culture--is available . The multifactorial problem of demystification is a difficult but not an impossible task . Less sensationalism, more real progress in research, selecting the right priorities, achieving the "ultimate drug", shelter, food, shoes, soap and broom for every human on this planet constitute the road to demystification.

Rev Pneumol Clin, 1990, 46(2), 49 - 54
{Intestinal perforation occurring at the beginning of treatment: a severe complication of bacillary tuberculosis}; Dore P et al.; We report 2 cases of intestinal perforation caused by tuberculosis and affecting the small intestine in one case and the colon in the other case . The patients were men aged 49 and 51 years respectively . Both were cachectic and presented with advanced open pulmonary tuberculosis . Perforation in free peritoneal cavity occurred 2 and 8 days respectively after an antituberculous treatment was initiated . The outcome was rapidly fatal in both cases . Tuberculous enteritis has become rare, but it can still be observed in patients with severe open pulmonary tuberculosis, where the gastro-intestinal tract is contaminated by the large number of virulent mycobacteria swallowed . In such patients clinicians must be alert to abdominal premonitory signs . Intestinal perforations in free peritoneal cavity are uncommon . Most perforation are small, single or multiple, and located on the antimesenteric side of the terminal ileum . They may occur at any time, and particularly just after an antituberculous therapy has been instituted . Clinical presentation is one of acute peritonitis requiring emergency laparotomy . Mortality has been reduced by technical improvements, notably temporary enterostomy, but perforation remains a serious and often fatal complication of tuberculosis in patients with severe malnutrition.

Rev Laryngol Otol Rhinol (Bord), 1990, 111(3), 251 - 3
{Efficacy of Cefsulodin in pyocyanic otologic infections, apropos of 55 cases}; Ennouri A et al.; Our work consists of a comparative study of 4 therapeutic protocols for treatment of otological infection from the pyocyanic bacillus, and more specifically of 19 patients with a necrotizing malignant otitis, 24 patients with perichondritis and 12 patients suffering from a post-operative infection . The association of Cefsulodin (a 3rd generation Cephalosporin) and Gentamicin gave the best results . A major decrease of the time of treatment and a total absence of infectious complications were in fact recorded . The adjuvant therapies were duly observed for the 4 protocols, in particularly the surgical treatment and the treatment of the site (diabetes).

Bull Soc Pathol Exot, 1990, 83(3), 376 - 93
{Agriculture-health interface in the field of epidemiology of vector-borne diseases and the control of vectors}; Mouchet J et al.; This paper is a review of the interactions between agriculture and vector borne diseases . Rain forest clearing makes possible the development of heliophilous species of anophelines and snails leading to an increase of malaria and schistosomiasis in Africa . But in Asia, clearing is a control method against Anopheles balabacensis, an important malaria vector . Clearing of forest galleries is followed by the disappearance of shore-dwelling tsetse flies . Woodcutters and pioneer farmers are contaminated with arbovirus and leishmaniasis when entering in natural sylvatic foci of these diseases . Management of drinking water reduces guinea worm as well as cholera and other diarrhoeal diseases . More over when piped water becomes available people are no more obliged to store drinking water in containers where vectors use to breed . Reservoirs of dams offer large possibilities for the development of mosquitoes including anophelines vectors of malaria and filariasis and of snails hosts of schistosomiasis . The medical importance of these man-made breeding sites depends of the local epidemiological features of the diseases . Dam spillways provide breeding for blackflies and man-made foci of onchocerciasis have been described in West Africa . Irrigation channels mainly when non cleared of vegetation are good breeding places for anophelines and snails . Irrigated surfaces like rice fields are highly productive in anophelines and other dangerous species of Culicinae . Insecticides used in agriculture, mainly to control cotton and rice pests, have been at the origin of insecticide resistance of several anopheline species . On an other hand, sometimes rice pests control lead to the control of rice field mosquitoes until they become resistant, e.g . for Culex tritaeniorhynchus the vector of Japanese encephalitis in South Korea . Many international organizations have emphasized the role of intersectorial collaboration to control man-made vector borne diseases foci . Good planning of the infrastructures (e.g . twin spillways) and adequate maintenance are essential . Vector control in rice field is a puzzling question . Wet irrigation was a hope but it cannot be done everywhere . Biological control methods have not been proven to be very efficient . Even Bacillus thuringiensis H14 and B . sphaericus have severe limitation . New tools for intersectorial activities should be a goal for scientists imagination.

Clin Ther, 1990, 12 Suppl C, 31 - 44
Gangrenous and perforated appendicitis with peritonitis: treatment and bacteriology; Bennion RS et al.; A comparison of single-agent antimicrobial therapy in the treatment of patients with perforated or gangrenous appendicitis and peritonitis was performed in a double-blind, randomized, prospective trial . Pathologic documentation of advanced appendicitis and positive intraoperative specimen cultures were required for inclusion in the study . Ceftizoxime (2 gm every 12 hours) and cefoxitin (2 gm every six hours) were compared . There were no significant differences between the treatment groups . Ninety-seven percent of patients treated with ceftizoxime and 89% of those treated with cefoxitin were cured or improved; there was no mortality in either group . By the use of optimal sampling, transport, and culture techniques, the number and diversity of bacteria recovered from these patients with advanced appendicitis were found to be much larger than previously suspected . Peritoneal fluid, abscess contents (if present), and appendiceal tissue (obtained so as to exclude the lumen) were cultured from all patients . An average number of 3.1 aerobic or facultative bacteria species and 8.5 anaerobic species were isolated from each specimen . Twenty-eight different genera and more than 55 species were encountered, including a previously undescribed fastidious gram-negative anaerobic bacillus . Bacteroides fragilis group and Escherichia coli were isolated from almost all specimens, and within the B fragilis group, eight species were represented . The recovery of such an unexpectedly large and diverse flora may be the reason for the therapeutic failures in these patients . We conclude that single-agent antimicrobial therapy in patients with advanced appendicitis and peritonitis is both safe and effective, and, with ceftizoxime, can be accomplished by a twice-daily dosing regimen.

Cancer Immunol Immunother, 1990, 31(5), 273 - 7
Idiotype vaccine for tumor by anti-idiotypic antibody prepared against anti-(bacillus Calmette Guèrin)BCG monoclonal antibody; Sasaki J et al.; The anti-idiotypic antibody (Ab2) prepared against the anti-BCG monoclonal antibody (mAb) (Ab1) exhibited potential vaccine activity against Meth A fibrosarcoma that shared a common antigen(s) with Mycobacterium bovis strain bacillus Calmette Guerin (BCG) . Mice vaccinated with the anti-idiotypic antibody (Ab2) were protected significantly against growth of the transplanted Meth A tumor (66%), and the presence of anti-(anti-idiotypic antibody) (Ab3) was proved in the Ab2-vaccinated mice by enzyme-linked immunosorbent assay and indirect immunofluorescence analyses using unabsorbed or absorbed sera against the BCG antigen(s) and Meth A tumor cells . This indicated that the anti-idiotypic antibody (Ab2) mimicked the structures of the BCG antigen(s) and behaved as the BCG antigen(s) to induce the Ab1-like antibody (Ab3) in vivo . Presumably the Ab2-induced Ab3 plays a significant role in preventing growth of the transplanted tumor in animals . By contrast, the control mice treated with normal mouse serum failed to inhibit the tumor growth . These results suggest the possible development of a tumor vaccine from the anti-idiotypic antibody (Ab2) prepared against the anti-BCG monoclonal antibody, for tumors sharing a common antigen(s) with Mycobacterium bovis strain BCG.

Antonie Van Leeuwenhoek, 1990 Jan, 57(1), 55 - 8
The problem of the taxonomy of the fusiform bacillus of acute necrotizing ulcerative gingivitis (Vincent's gingivitis); Holbrook WP et al.; The characteristic microscopic appearance of the fusospirochaetal complex in smears from patients with acute necrotizing ulcerative gingivitis has long been regarded as virtually pathognomonic of the disease . Unfortunately there is considerable confusion surrounding the taxonomy of the fusiform organism in the fusospirochaetal complex . The conflicting views appear to be irreconcilable and a re-evaluation of the taxonomy of oral fusiforms is urgently required.

Ann N Y Acad Sci, 1990, 585, 386 - 93
Thermostable alanine racemase . Its structural stability; Soda K et al.; The gene encoding thermostable alanine recemase from Bacillus stearothermophilus was cloned and expressed in E . coli . The enzyme was purified to homogeneity from cell extracts of E . coli carrying a plasmid designated pICR4 . The alanine racemase gene sequenced was found to contain an open reading frame of 1158 nucleotides . The molecular weight of the enzyme subunit was estimated to be 43,341 . The alpha-helical and beta-structure contents were calculated to be about 34 and 26%, respectively, from CD data . CD measurements of the denaturation process of enzyme by guanidine hydrochloride showed the presence of a stable intermediate during the denaturation . Limited proteolysis with subtilisin resulted in the formation of two dissimilar peptide fragments with molecular weights of about 28,000 and 13,000 in the early stage of the digestion . These suggest that the enzyme subunit is composed of two structurally dissimilar domains connected by a short polypeptide (residues 258-266), which first suffers the limited proteolysis . However, the enzyme retained almost full activity and the conformation indistinguishable from the intact protein even when it was proteolytically hydrolyzed to more than 10 fragments.

Urol Int, 1990, 45(3), 137 - 41
Intravesical Bacillus Calmette-Guérin prophylactic treatment for superficial bladder tumors: results of a controlled prospective study; Melekos MD; A controlled prospective study in 100 patients evaluated the efficacy of intravesical bacillus Calmette-Guerin (BCG) administration as prophylactic treatment on tumor recurrence and tumor progression rate after endoscopic resection of superficial transitional cell carcinoma of the bladder . There were 27 recurrences in 22 of 67 evaluable patients (33%) who received BCG, compared to 27 recurrences in 19 of 33 control patients (58%) (p less than 0.05) . The mean follow-up periods for the tumor-free patients in the BCG and control groups were 29 and 30 months, respectively, while the mean times to tumor recurrences for the above groups were 13.36 +/- 6 and 9.94 +/- 5 months, respectively (p less than 0.05) . The recurrence rates per 100 patient months for the BCG and control patients were 1.69 and 4.41 recurrences, respectively (p less than 0.05), while 7 patients of the BCG group showed recurrent tumors of higher stage or grade, compared to 13 of the controls (p less than 0.05) . This study confirms that Pasteur strain BCG is safe and efficacious in the prevention of superficial bladder tumor recurrence and tumor progression.

Urol Int, 1990, 45(3), 129 - 36
Use of intravesical bacillus Calmette-Guérin in the treatment of superficial transitional cell carcinoma of the bladder: an overview; Witjes JA et al.; Since 1976 superficial bladder cancer is treated with intravesical instillations of Bacillus Calmette-Guerin (BCG) . The results with BCG are superior to other intravesical chemotherapeutics, both in superficial tumors and carcinoma in situ . BCG probably acts as a nonspecific potentiator of the immune system . Comparative studies have to give information about the effectiveness and side effects of the different strains of BCG, and about the best clinical schedule . Toxicity is mild in 95% of the cases . Severe side effects and antituberculous drugs are discussed.

J Basic Microbiol, 1990, 30(4), 251 - 8
Bacillus thuringiensis insecticidal delta-endotoxin: diversity of crystal proteins and its relatedness to the toxicity spectrum; Haider MZ et al.; Bacillus thuringiensis strains produce crystal delta-endotoxins which exhibit a diverse toxicity spectrum . In order to explore the basis of toxin specificity, a comparison of the activity of 13 strains belonging to seven serotypes was made against three insect species . The delta-endotoxin crystals were purified and their polypeptide composition analyzed by SDS-PAGE . Among the strains studied, the delta-endotoxins consist of a variety of crystal proteins in the 60-144 KDa size range . On the basis of molecular mass, endotoxins maybe grouped into two classes; one contained both high (125-144 KDa, P1) and medium sized (60-66 KDa, P2) proteins and a second class consisting of only the high Mr polypeptides . Immunoblotting with B . aizawai P1 antiserum revealed antigenic cross-reaction with one or more of the polypeptides in 125-144 KDa range in all the strains studied . When the crystal proteins from different strains were immunoblotted with kurstaki P2 antiserum, none of the P1 protein crossreacted suggesting that the P1 and P2 proteins are not structurally related . However, the B . kurstaki P2 antiserum crossreacted with 66 KDa proteins in some other strains which underlines a structural homology in this class of the toxic polypeptides . Toxicity studies revealed that the high Mr P1 proteins of all the strains in this study were active against lepidopteran (Pieris brassicae and Diacrisia obliqua) larvae . B . thuringiensis aizawai strains exhibit a dual toxicity associated with the high Mr (130-135 KDa; P1) proteins . The P2 crystal proteins (60-66 KDa) also showed dual toxicity against the lepidopteran and dipteran larvae but were found to be structurally and immunologically distinct.

Biochem Int, 1990, 20(3), 607 - 13
Human placental carboxypeptidase M is anchored by a glycosyl-phosphatidylinositol moiety; Shimamori Y et al.; Basic carboxypeptidase activity was released from human placental membranes on treatment with phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis . The enzyme was successively purified to homogeneity by SDS-polyacrylamide gel electrophoresis . The molecular nature and some catalytic properties of the purified enzyme revealed that it is identical with recently described basic carboxypeptidase M (R.A . Skidgel et al . J . Biol . Chem . 264 (4) 1989 2236-2241).

FEMS Microbiol Lett, 1990 Jan 1, 54(1-3), 277 - 9
Suppression of tyrocidine production by purine nucleotides and related substances in Bacillus brevis; Oyama M et al.; Bacillus brevis (ATCC 8185) produces an antibiotic peptide, tyrocidine . We found that adenosine or 5'-AMP suppressed the production of tyrocidine with half-maximum inhibition at 100-300 microM . This inhibition was specific to the production of tyrocidine since neither adenosine nor 5'-AMP showed any effect on bacterial growth . Cyclic nucleotides had no effect . These results suggest that adenosine, 5'-AMP or its metabolite was specifically involved in the regulation of tyrocidine production.

Appl Environ Microbiol, 1990 Jan, 56(1), 162 - 6
Toxicity of protease-resistant domains from the delta-endotoxin of Bacillus thuringiensis subsp . israelensis in Culex quinquefasciatus and Aedes aegypti bioassays; Pfannenstiel MA et al.; The mosquitocidal glycoprotein endotoxin of Bacillus thuringiensis subsp . israelensis was digested with chymotrypsin to yield protease-resistant domains which were then separated from smaller protease digestion products by high-performance liquid chromatography . Once purified, the domains no longer bound wheat germ agglutinin, a lectin which binds N-acetylglucosamine (GlcNAc) and GlcNAc oligomers . Purified protease-resistant domains were as toxic for Culex quinquefasciatus larvae as intact solubilized toxin . In separate experiments, the toxicity of chymotrypsin-digested endotoxin for Aedes aegypti larvae was reduced fivefold or more . A model is presented in which GlcNAc-containing oligosaccharides are required for toxicity for A . aegypti larvae but not C . quinquefasciatus larvae.

Clin Exp Immunol, 1990 Jan, 79(1), 7 - 14
A complex component modulating immune-deficient cells in leprosy patients leading to loss of viability of Mycobacterium leprae--a possible vaccine; Marolia J et al.; Macrophages from peripheral blood of leprosy patients, both multi-bacillary and paucibacillary are unable to kill phagocytosed Mycobacterium leprae due to their inability to produce superoxide (O2-) and hydroxyl radicals (OH.) . The macrophages from healthy individuals are able to kill M . leprae along with release of O2- and OH . radicals . The deficiency in the macrophages of both types of leprosy patients is removed by activation of these cells when exposed to a culture supernatant obtained after stimulation of peripheral blood mononuclear cells from the same patients with delipidified cell components of M . leprae which are most likely cell wall proteins . The activation of macrophages also leads to recognition of whole live M . leprae as an antigen by cells from lepromatous patients . This activation of the phagocytes by delipidified cell components is blocked by cyclosporin A, indicating the possible role of several steps involved in immune activation of cells . The observations thus indicate the significant ability of delipidified cell components to eliminate the deficiencies in the macrophages from leprosy patients and restore them to behave like the cells from healthy individuals . Considering all these, it is suggested that delipidified cell components could be potential modulators, and are probably capable of functioning as a vaccine for leprosy.

Immunopharmacol Immunotoxicol, 1990, 12(4), 583 - 94
Efficacy of immunopriming prior to isolation of tumor infiltrating lymphocytes for use in adoptive immunotherapy; Rubenstein M et al.; Adoptive immunotherapy is dependent upon the leukocytic subsets isolated as tumor infiltrating lymphocytes (TIL) prior to in vitro expansion with interleukin-2 . To favorably influence T-cell subset representation in TIL the efficacy of bacillus Calmette Guerin (BCG) and cyclophosphamide (CTX) priming was evaluated in rats bearing Dunning R3327-AT prostatic tumors . When assessed by immunohistochemistry, both agents significantly (p less than 0.001) increased helper-T representation and decreased that by suppressor-T cells . As a result helper/suppressor (H/S) T cell ratios of TIL from untreated tumors (0.73 +/- 0.11) were significantly (p less than 0.001) elevated by both BCG (1.93 +/- 0.39) and CTX (1.40 +/- 0.25) . Immunopriming might enhance adoptive immunotherapy by increasing the H/S ratio of TIL prior to their culture.

Probl Tuberk, 1990, (10), 17 - 20
{Results of ambulatory observation of patients registered as group O in an anti-tuberculosis dispensary}; Dzhunusbekov AD et al.; 414 patients registered under O-group, 149-under VIIB and 129-under IA were observed by the antituberculosis dispensary of Alma-Ata . The minor differences detected in the clinical symptomatology of these patients did not allow one to decide how the process was proceeding in a specific case . The results showed that it was advisable to enroll++ subjects not with focal, but with cirrhotic and disseminated processes, tuberculomas and intrathoracic lymphatic tuberculosis into the O-group . The presence of X-ray archiv does not rule the necessity of following-up in this group in some cases . The process activity in the O-group can be defined by such methods, as bacillary excretion test and study of X-ray dynamics in the presence of challenge therapy, while in cirrhoses and intrathoracic lymphatic tuberculosis, only bacillary excretion test can be done . The outcomes of observation, in addition traditional ones (transfer to IA and VIIB groups), may lead to the transfer to VIIA group and striking off the register of the antituberculosis dispensary . Direct and indirect economic impact of the observation in the O-group of the dispensary registration was estimated.

Probl Tuberk, 1990, (9), 21 - 4
{Tuberculosis infection and morbidity in children of large and small families in relation to epidemic risks of tuberculosis foci}; Murriev A; Tuberculosis contamination, primary infection and morbidity rates in 1,162 children from 484 family foci of the infection in relation to epidemic hazard of the affected and the number of children in contact with the latter were studied . Contamination of children in large families is 1.4 times higher than in small ones (61.6 and 44.7%, respectively); primary infection is 1.5 times higher (30.2 and 19.8%); and morbidity rate is 2.3 times higher (22.7% in large and 9.8% in small families per 1000 man/years of observation) . In this case a direct relation of the main epidemiological characteristics of tuberculosis to the degree of the bacillary excretion intensity, duration and nature of the contact, number of children in a family, etc . is found . Therefore, large family foci of the tuberculous infection should be referred to Group 1 according to the accepted classification of tuberculosis foci irrespective of either presence or absence of bacillary excretion or its intensity.

Z Versuchstierkd, 1990, 33(4), 163 - 9
Epidemiological studies of Bacillus piliformis infection and Tyzzer's disease in laboratory rats; Hansen AK et al.; Characteristics in connection with the presence of Bacillus piliformis in the barrier-protected animal rooms of a breeding centre for laboratory rats are discussed . The incidence of antibodies against B . piliformis in IFA test is high -77.8% for the total breeding centre . Clinical and pathological changes are, however, rare and mainly connected with certain rat strains . Immunosuppression with prednisolone and subsequent histopathological tests of liver sections indicate that rats with MHC-type RT1.A-u have an increased risk of developing Tyzzer's disease, while RT1.A-1-carriers seem to be resistant to this disease.

Allergol Immunopathol (Madr), 1990 Jan-Feb, 18(1), 47 - 52
Evaluation of circulating immune complexes in cutaneous diseases associated with immune disorders; Fernandez Bussy R et al.; Several skin diseases associated with immune disorders may be related to the formation of circulating immune complexes (CIC) and their skin deposition . Sera from 20 controls and 108 patients, including: 23 psoriasis, 10 lichen planus, 30 atopic eczemas, 32 cases of leprosy, 10 vasculitis and 3 pyoderma gangrenosum were evaluated for the presence of IgG-containing CIC by the microconsumption complement test (MCT) . Additionally, the presence of IgE-containing CIC by means of a polyethylene glycol precipitation and radioimmunoassay technique was evaluated in 10 patients with atopic eczema . It was found that 56.5% of psoriatic patients show moderate CIC concentrations, as well as 34% of leprosy patients, with increased levels when bacillus were detected in skin lesions, and in 90% of leukocytoclastic vasculitis . A close relationship between CIC levels and the clinical evolution of skin lesions was demonstrated in patients with pyoderma gangrenosum and lichen planus . IgG-CIC were detected in 33% and IgE-CIC in 30% of patients with atopic eczema, with the presence of both types of CIC in 2 out of 3 cases . The systematic research on CIC presence in some selected skin diseases shows that immune complexes take part, with different degrees of relevance, in the pathogenesis of all the entities.

Toxicol Appl Pharmacol, 1990 Jan, 102(1), 21 - 33
Effects of ozone, hexachlorobenzene, and bis(tri-n-butyltin)oxide on natural killer activity in the rat lung; Van Loveren H et al.; The respiratory tract is a major route of exposure to noxious agents as well as pathogens such as viruses . Natural killer (NK) activity is an important first line of defense to virally infected cells as well as certain neoplasms; therefore, testing the effects of exposure to toxic compounds on this activity is important in understanding the immunotoxic potential of the compound . Lymphoid cell suspensions, obtained after enzymatic dispersion of rat lungs and purification over nylon wool columns, showed in vitro natural killer activity toward YAC lymphoma cells . Validation of the test with well-known NK activity stimulators such as Bacillus Calmette-Guerin (BCG), interleukin-2 (IL-2), interferon (IFN), and inhibitors like anti-asialo-GM1 (ganglio-n-tetrasylceramide) antibody confirmed the reliability of the test as an assay for detecting NK activity in rat lungs . Using this assay, we studied the effects of exposure to ozone (O3), hexachlorobenzene (HCB), and bis(tri-n-butyltin)oxide (TBTO) on NK activity in rat lung . Inhalation exposure to O3 for 7 days at 0.4 and 0.8 mg/m3 resulted in stimulation, and exposure at 1.6 mg O3/m3 resulted in suppression of NK activity . Oral exposure to HCB in concentrations of 150 and 450 mg/kg food for 6 weeks suppressed NK activity in rat lungs in a dose-related manner . This was also true for 6 weeks of oral exposure of rats to 20 and 80 mg TBTO/kg food, but to a lesser extent . In summary, we have developed and validated a method to measure the effects of (toxic) substances on NK activity in rat lung.

J Egypt Public Health Assoc, 1990, 65(3-4), 319 - 34
Antibacteriophage action on the larvicidal activity of Bacillus thuringiensis H-14 and Bacillus sphaericus against Culex pipiens; Rady MH et al.; An organically polluted mosquito breeding water was tested for the presence of bacteriophages which could inhibit the larvicidal activity of B . thuringiensis H-14 and B . sphaericus . More than one bacteriophage were isolated which could inhibit the tested bacteria . The sensitivity of the two bacterial species to 12 antibiotics was tested . Two of them, showed no antibacterial action, were selected and considered as antivirus agents in the bacteriophage assays; namely, Amoxycillin and Co-Trimoxazole . Results indicated their antivirus activity as the addition of the antibiotics to the isolated phages could permit normal bacterial growth as well as persistence of larvicidal activity against Culex pipiens larvae.

Rocz Panstw Zakl Hig, 1990, 41(5-6), 269 - 72
{Sensitivity to antibiotics and selected compounds of Bacillus cereus isolated from food products and cases of food poisoning}; Stec E; The study was done on 60 strains of Bacillus cereus isolated from cases of food poisoning and from food products tested in current supervision . The highest sensitivity was found to the antibiotics: streptomycin, neomycin, gentamicin, amikacin, oxytetracycline, and sulphonamides: negram and sulfathiazole . All strains were resistant to colistin and cloxacillin, nearly all were resistant to penicillin, ampicillin and optochine.

Probl Tuberk, 1990, (12), 5 - 8
{Clinical value of detection of L forms of Mycobacterium tuberculosis in patients with residual tuberculous changes in the lungs}; Dorozhkova IR et al.; To clarify the role of M . tuberculosis L-form in a recurring tuberculous process in subjects with residual tuberculous changes in the lungs, microbiological, ++clinico-roentgenological and laboratory examinations of 1651 persons recorded as having VIIA, VIIB, III and 0 groups were undertaken . The causative agent of tuberculosis was isolated in a typical bacillary and L-transformed forms in 3.1 and 5.0% of the screened persons, respectively . Active respiratory tuberculosis was diagnosed in 64.7% of sputum-positive patients, including 86.3% of them as bacillary carriers and 42.7% of those excreting L-forms of the tuberculosis agent . The detection of biologically altered forms of the causative agent indicates a potential activity of the process and requires an intensified medical examination control and differential prevention activities.

Genetica, 1990, 81(3), 215 - 20
High temperature of development and selection of Bacillus thuringiensis-supernatant-resistant females in a Drosophila melanogaster Oregon R strain; Paumard-Rigal S et al.; The supernatant of Bacillus thuringiensis cultures contains a thermostable toxin: the beta exotoxin or thuringiensin, which in vivo acts as a preferential inhibitor of ribosomal RNA synthesis . Added to Drosophila melanogaster culture medium, it induces, during the flies' development, a lethal effect that is, in our Oregon R strain, greater for females than for males . The authors have previously shown that a diminution of the ribosomal DNA amount increases the sensitivity to the lethal effect of the supernatant . From a stock subjected for several generations to a 28 degrees C temperature and more, they have spontaneously obtained a variant population where males and females have similar resistance . It is shown here that this difference between the two populations is expressed in the X/X female genotypes, and that there is a correlation between the sensitivity to the lethal effect of the Berliner Bacillus thuringiensis supernatant and the sensitivity to the effect of the 28 degrees C developmental temperature; genotypes resistant to the lethal effect of the temperature were positively selected when the temperature of development was increased . The better resistance of these genotypes could be related to more active ribosomal units on the X chromosome.

Biol Met, 1990, 3(3-4), 188 - 96
Mercury resistance determined by a self-transmissible plasmid in Bacillus cereus 5; Belliveau BH et al.; Inducible mercuric reductase activity in Bacillus cereus 5 was plasmid-encoded . Plasmid analysis revealed three plasmids with molecular masses of 2.6, 5.2 and 130 MDa . A mating system permitted transfer of the resistance determinant among strains of B . cereus and B . thuringiensis . Transfer of mercury resistance from B . cereus 5 to B . cereus 569 and B . thuringiensis occurred during mixed culture incubation on agar surfaces . The 130-MDa plasmid (pGB130) was responsible for transfer; frequencies ranged from 10(-5) to 10(-4) . B . cereus 569 transconjugants inheriting pGB130 were also effective donors . High transfer frequencies and the finding that cell-free filtrates of donor cultures were ineffective in mediating transfer suggested mercury-resistance transfer was not phage-mediated . Transfer was also insensitive to DNase activity . Further evidence that pGB130 DNA carried the mercury-resistance determinant was transformation of B . cereus 569 by electroporation with pGB130 DNA isolated from B . cereus 5 and a mercury-resistant B . cereus 569 transconjugant . Mercury-resistant transconjugants and transformants exhibited mercuric reductase activity . Plasmid pGB130 also conferred resistance to phenylmercuric acetate.

Folia Microbiol (Praha), 1990, 35(5), 402 - 12
Characteristics of intracellular proteolytic activities of Bacillus megaterium; Moravcova J et al.; Intracellular proteolytic activities of B . megaterium KM occur soluble in the cytoplasm and periplasm and insoluble in the membrane . Two proteolytic enzymes were found in the cytoplasmic fraction by gel filtration on Sephadex G 150 and by polyacrylamide gel electrophoresis . The first enzyme called CI was stable, had a relative molecular mass of Mr = 105,000 (M = 105 kg/mol) and was inhibited by EDTA and PMSF, whereas the second, designated CII, was labile and had a relative molecular mass of Mr = 46,000 (M = 46 kg/mol) . Because of its lability it could not be characterized in detail . In the "periplasm" only a single proteolytic enzyme P (Mr = 28,000; M = 28 kg/mol) inhibited by EDTA could be demonstrated . The extracellular enzyme exhibited similar properties . The membrane proteolytic activity was sensitive to PMSF and EDTA . The membrane enzymes have not yet been solubilized . In cells of the mutant KM 12 that does not produce the extracellular proteinase, only one type of proteinase, in all its properties identical with the cytoplasmic proteinase CI, could be demonstrated.

Rocz Panstw Zakl Hig, 1990, 41(3-4), 194 - 9
{Intensity of hemolysin and lecithinase production by Bacillus cereus strains isolated from food}; Stec E; The study was carried out on the filtrates of cultures of 60 Bacillus cereus strains isolated from food products which caused food poisoning (20 strains), from patients (20 strains) and from food analysed in routine supervision . The strains isolated from food poisoning demonstrated a more pronounced haemolytic activity as compared with the strains from routine supervision . A lack of correlation was demonstrated between the intensity of lecithinase production and the intensity of reaction of rabbit ileum loop . No strict correlation was noted between the haemolytic properties and lecithinase activity.

Folia Microbiol (Praha), 1990, 35(3), 190 - 9
Effect of actinomycin D on viability, sporulation and nucleotide pool of Bacillus megaterium; Vachova L et al.; A transient 7-fold rise of ppGpp concentration, 2-3-fold increase of pppGpp concentration and 50% drop of the concentration of GTP in Bacillus megaterium cells immediately after their transfer to the sporulation medium were observed . Actinomycin D, in concentrations inhibiting RNA synthesis by 95%, blocked the rise of the (p)ppGpp pool and caused an instant several-fold increase of the GTP level . When the cells were exposed to actinomycin D in the sporulation medium for a 1-h period (time 0-1 h, 1-2 h or 2.20-3.20-h), they were able to form colonies on nutrient agar after being kept, in addition for 1-2 h in the sporulation medium free of the antibiotic . The ability of sporulation was, however, markedly limited . The share of cells that could sporulate increased when the irreversible sporulation phase was reached.

Probl Tuberk, 1990, (5), 48 - 50
{Characteristics of bacterial excretion in conio-tuberculosis of the lungs}; Noreiko BV et al.; Bacteriological examination of sputum smears of 42 patients with anthracotic tuberculosis divided according to the activity of a tuberculous process into 2 groups was undertaken . To achieve the growth of L-forms, sputum inoculation was done on Shkolnikova's semiliquid culture medium . Phase contrast microscopic examination of the sputum smears was carried out at 280 X magnification . As the pneumoconiotic process in patients with anthracotic pulmonary tuberculosis grows progressively worse, the rate of typical Mycobacterium strain positivity decreases, while the rate and intensity of L-forms excretion proportionally increase . In the patients with inactive forms of anthracotic tuberculosis, the bacillary excretion was characterized by monomorphism of vacuolized L-forms . At the same time, active forms of anthracotic tuberculosis were accompanied by a massive excretion of Myco . L-forms and featured a wide range of morphologic changes dominated by giant cells, cord-factor, heterogeneous granulation and homogeneous mass as a consequence of overall destruction of the cell membranes.

Int Arch Allergy Appl Immunol, 1990, 91(3), 278 - 84
Antigenic and allergenic characterization of the enzymes alcalase and savinase by crossed immunoelectrophoresis and crossed radioimmunoelectrophoresis; Arlian LG et al.; Alcalase and savinase, produced by Bacillus species, are proteolytic enzymes that are used in laundry products and are known to cause respiratory allergy . Antigenic and allergenic characteristics of alcalase and savinase and their potential cross-reactivity were evaluated using crossed immunoelectrophoresis and crossed radioimmunoelectrophoresis . Alcalase exhibited two distinct antigens; one electropositive and one electronegative . The electropositive antigen exhibited some retrograde anodic mobility when coupled with antiserum components . Savinase exhibited one electropositive and two electronegative antigens . The antigens of the two enzymes were clearly different from each other, the three savinase antigens exhibiting greater electrophoretic mobility than the two alcalase antigens . In crossed radioimmunoelectrophoresis studies, only the electropositive antigen of alcalase, its retrograde complex, and the electropositive antigen of savinase bound IgE from the sera of individuals who were skin test positive to one or both enzymes . No evidence of cross-reactivity was observed in heterologous and tandem crossed immunoelectrophoresis studies and heterologous microimmunodiffusion reactions.

Prikl Biokhim Mikrobiol, 1990 Jan-Feb, 26(1), 61 - 6
{Dynamics of accumulation of extracellular lipoteichoic acid in Bacillus cereus}; Pronin SV et al.; The dynamic of accumulation of extracellular lipoteichoic acid (LTA) was studied depending on the growth stage of Bacillus cereus st . 96 . A maximum amount of extracellular LTA was detected in the middle of the exponential growth . The quantity of the biopolymer present in the culture medium at the beginning of the stationary growth under conditions of catabolite repression of sporulation and without repression was found to be different . Experimentally increased concentrations of LTA inhibited B . cereus sporulation . Besides, dormant spores of B . cereus st . 96 were found to contain LTA.

Biokhimiia, 1990 Jan, 55(1), 87 - 94
{Characteristics of kinetics of phospholipid hydrolysis by phospholipase C from Bacillus cereus . Hydrolysis of phosphatidylcholine in the presence of deoxycholate}; Voronin MV et al.; Using dynamic light scattering and 31P-NMR spectroscopy methods, the reaction of solubilization of phosphatidylcholine by the ionic detergent, sodium deoxycholate, in aqueous solutions was studied . The kinetics of phosphatidylchodine hydrolysis by phospholipase C from B . cereus depending on the size and structural organization of substrate aggregates was investigated . No phosphatidylcholine hydrolysis was observed in the case of lamellar organization of the substrate, the size of lamellas not exceeding 2000-5000 A . The substrate hydrolysis rate within mixed micelles was controlled by the accessibility of the substrate on the surface of micellar aggregates . There was a decrease in the phosphatidylcholine hydrolysis rate at high detergent concentrations in the system . It was concluded that such a decrease in the hydrolysis rate can be due to two reasons, i) the decrease in mixed micelle size with a simultaneous decrease of surface concentration of the substrate, and, ii) the formation of "pure" detergent micelles capable to adsorb the enzyme by decreasing the "effective" concentration of phospholipase C.

J Appl Bacteriol, 1990 Jan, 68(1), 17 - 21
Isolation and propagation of phages naturally associated with the aizawai variety of Bacillus thuringiensis; Inal JR et al.; This study describes the isolation of a phage, using mitomycin C and u.v . light, from each of four strains (HD67, HD130, HD228 and HD248) of Bacillus thuringiensis H-serotype 7 (var . aizawai) . It also describes the isolation of two indicator strains (12.13 and HD102) for these phages (phi HD67, phi HD130, phi HD228 and phi HD248) and the ideal conditions, using these indicator strains, for maximum phage production.

Br J Ophthalmol, 1990 Jan, 74(1), 26 - 9
Bacillus-induced endophthalmitis: new series of 10 cases and review of the literature; Hemady R et al.; We reviewed the charts of 10 patients who were admitted to the Massachusetts Eye and Ear Infirmary over a 10-year period with the diagnosis of Bacillus species endophthalmitis . To our knowledge this is the largest single series in the literature and includes the first two reported cases of Bacillus endophthalmitis following glaucoma filtering procedures . Seven cases developed following penetrating ocular trauma . One occurred in an intravenous drug abuser . Five eyes ultimately underwent enucleation; only the two eyes that developed endophthalmitis after elective surgery retained useful vision . Review of the literature indicates that parenteral and intravitreal antibiotic prophylaxis against endophthalmitis after penetrating ocular trauma should include gentamicin, in combination with vancomycin or clindamycin, to provide adequate coverage against infection with Bacillus spp., as prognosis is poor once infection is established . Bacillus spp . cultured from ocular tissues or fluids should not be dismissed as contaminants.

J Urol, 1990 Jan, 143(1), 34 - 5; discussion 35-6
Topical mitomycin C therapy for carcinoma in situ of the bladder: a followup; Stricker PD et al.; We studied 15 patients with histologically proved multifocal carcinoma in situ of the bladder who were in remission at a mean followup of 21 months after induction intravesical chemotherapy with mitomycin C . These patients have been followed for a further 28 months, for a total mean duration of 49 months . Of the 15 patients 4 suffered new areas of carcinoma in situ, including 3 who subsequently required cystectomy (2 after unsuccessful intravesical bacillus Calmette-Guerin therapy and 1 with a simultaneous invasive tumor) . One patient underwent transurethral resection of the prostate for carcinoma in situ of the prostatic urethra, which subsequently was shown to be limited to mucosa and not involving the deeper ducts nor the stroma . Of the remaining 11 patients 1 died of unrelated disease and 2 suffered recurrent papillary transitional cell carcinoma treated successfully with a combination of intravesical bacillus Calmette-Guerin therapy and resection . The other 8 patients have remained free of tumor . None of the 15 patients had metastatic cancer . We believe that these results support the durability of response after induction mitomycin C therapy . We stress the necessity for prolonged close followup to detect recurrent tumor and to avoid metastatic disease.

Chin J Biotechnol, 1990, 6(3), 173 - 7
Cloning and expression of mosquito larvicidal protein gene from a highly toxic local strain of Bacillus sphaericus; Liu Y et al.; Bacillus sphaericus strain 10 (BS10), isolated from Jiangsu Province of China, is highly toxic to the mosquito larvae . During sporulation, BS10 produces a parasporal crystalline protein which is toxic to the larvae of a number of mosquito species, and extremely toxic against the larvae of Culex pipiens . Using the Escherichia coli cloning vector pAT153, two clones which hybridized with the synthesizing 18-bases oligonucleotides probe have been obtained . One of the recombinants, TG1 (pFL37), contains a 4.0 kb HindIII DNA fragment from BS10, and produces the 43 kd larvicidal toxin protein . Western blotting analysis and biological assay of mosquito larvicidal activity confirm that larvicidal toxin gene of BS10 is expressed in E . coli.

Nouv Rev Fr Hematol, 1990, 32(6), 461 - 3
Cat-scratch disease bacteria; Arlet G et al.; Cat-scratch disease is a benign inoculative lymphoreticulosis, related to the presence of a polymorph bacillus, Warthin-Stary silver stained, Gram negative as assessed by Brown-Hopp staining . It is found in the capillary walls and in macrophages bordering the lymph node sinusoids at the site of inoculation, in regional subacute adenopathy before softening, in internal organs and blood cultures of systemic infections, occurring more often in immuno-compromised patients . These bacteria have been demonstrated in subcutaneous vascular nodules, near to histiocytoid hemangioma in AIDS patients; these lesions are very similar to early stage Kaposi's sarcomas . This bacteria is provisionally listed as G 1492 by the Center for Disease Control.

Acta Microbiol Bulg, 1990, 26, 77 - 83
{The phage sensitivity and insecticidal activity of local strains of Bacillus sphaericus}; Grigorova R et al.; The sensitivity of 132 local isolates of B . sphaericus to phages 3, 4 and SST of A . Yousten has been studied . According to this incomplete phage typing, most of the strains are classified in the third phage group . A high-degree correlation between sensitivity to phage 4 and insecticidal activity was established, owing to which this sensitivity can be used as an initial criterion for selection of newly isolated strains . LC50 and LC90 of one reference and six local strains of B . sphaericus were determined using Culex pipiens third instar larvae, and the local strains were found to exceed in insecticidal activity the reference strain 2362 . This suggests that the newly isolated strains are promising for developing of a preparation for biological control of mosquitoes.

Scand J Infect Dis Suppl, 1990, 74, 235 - 48
Human pharmacodynamics of beta-lactams, aminoglycosides and their combination; Drusano GL; In-vitro and animal model data indicate that the time beta-lactam serum concentrations remain above the MIC is an important determinant of the organism kill at the primary infection site . Similarly, for aminoglycosides, area under the curve and peak concentrations have been linked to organism kill and suppression of resistance . It is only in clinical patients that these data can be validated as to their significance . For beta-lactams, little clinical data exist regarding these concepts . However, Bodey & colleagues have shown that profoundly, persistently neutropenic cancer patients fared better when one of their beta-lactams was administered continuously . Our group was able to correctly predict outcome in 9/10 patients bacteremic with a Gram-negative bacillus when receiving a single beta-lactam on the basis of the time free drug concentrations remained above the MIC . Schentag et al studied patients with lower respiratory tract infection treated with cefmenoxime alone and found a significant relationship between time greater than MIC & time to clearance of the pathogen from cultures of the tracheobronchial tree . These data would seem to validate the predictive nature of the findings from in-vitro & animal model studies . With aminoglycosides, Moore, Smith & Lietman were able to demonstrate a highly significant correlation between outcome and the maximal peak concentrations to MIC ratio achieved for patients with single organism Gram-negative rod infections . This is somewhat at variance with some animal models, but as the studies were performed with a fixed dosing interval, the outcome is not surprising . Little has been done with combinations of these agents in patients . Barriere & colleagues have proposed the AUC of the reciprocal serum bactericidal activity curve as a way to integrate the activity of combinations . We have developed a method employing logistic regression analysis to integrate the activity from the administration of multiple agents . The integration of this approach with each drug's pharmacokinetics allows the generation of a plot of the probability of the blood remaining sterile over a steady state dosing interval . This approach has been preliminarily tested in 6 individuals with excellent concordance between outcome and prediction . Development of data in-vitro and in animal models with validation in patients will hopefully provide the impetus to optimize therapy, and thence, outcome for the most seriously ill individuals.

Int Urol Nephrol, 1990, 22(5), 433 - 40
Intravesical bacillus Calmette-Guérin administration in the prophylaxis of superficial bladder cancer; Melekos MD et al.; A total of 68 patients received prophylactic/adjuvant intravesical Pasteur strain bacillus Calmette-Guerin (BCG) instillations following transurethral resection for superficial bladder carcinoma . An induction phase consisting of 6 weekly instillations was followed by a maintenance phase consisting of 1 instillation given every 3 months . A second 6-week course of BCG was administered to initial failure followed again by a quarterly maintenance therapy . The response rate for patients treated with one 6-week course was 56%, while complete response was achieved in 72% when both treatment courses were considered . Follow-up period lasted at least 2 years after each course of BCG . These results suggest that additional courses of BCG increase the prophylactic efficacy of intracavitary BCG . Regarding the purified protein derivative (PPD) skin test and granulomatous response in the bladder, it has been shown in the present study that, although favourable results occurred more frequently among patients with either PPD conversion from negative to positive or vesical granuloma formation, neither of these indices should be considered reliable prognostic indicators.

Rev Pneumol Clin, 1990, 46(5), 216 - 20
{Mediastinal tuberculous adenopathies in Djibouti . Report of 159 cases}; Auregan G et al.; One hundred and fifty-nine cases of confirmed mediastinal tuberculous adenopathy were analysed retrospectively . This series, which is the first reported in French-speaking Africa, shows that the disease is not uncommon there, and not restricted to black Africans living in Europe . The clinical symptoms are not specific . Radiography is more interesting, and the extreme scarcity of bacillys-containing expectoration encourages endoscopy . The finding of a fistula at fibroscopy confirms the diagnosis . This easy examination is particularly useful since M . tuberculosis can be grown in cultures from ground bronchial fistula biopsies in 86% of the cases . Associated tuberculous lesions in other sites are very frequent (45%), as is extension of the disease from lymph nodes to lung tissue (79%) . This clinical form of tuberculosis can be treated with modern short course chemotherapy which is remarkably effective.

Comp Biochem Physiol C, 1990, 95(2), 241 - 5
Effects of Bacillus thuringiensis delta-endotoxin on the permeability of brush border membrane vesicles from tobacco hornworm (Manduca sexta) midgut; Hendrickx K et al.; 1 . The effect of two recombinant Bacillus thuringiensis delta-endotoxins on brush border membrane vesicles of Manduca sexta midgut was investigated using an in vitro assay system, based on ion-amino acid cotransport . 2 . A CryIA(b)-toxin provoked an increase in the permeability of the vesicles . 3 . A CryIB-toxin, not toxic to M . sexta larvae in vivo, had no effect in our assay . 4 . In contrast to earlier reports, the increase in permeability was found to be neither selective for K+ nor specifically inhibited by Ca2+ or Ba2+ . 5 . Our data support the hypothesis that B . thuringiensis delta-endotoxins create non-specific pores.

FEMS Microbiol Lett, 1990 Jan 1, 54(1-3), 257 - 62
Development of mutants of the mosquitocidal bacterium Bacillus thuringiensis subspecies morrisoni (PG-14) toxic to lepidopterous or dipterous insects; Padua LE et al.; The parasporal body of the mosquitocidal isolate (PG-14) of Bacillus thuringiensis subsp . morrisoni (BTM) contains five major proteins with molecular masses of, respectively, 27.3, 65, 128, 135, and 144 kDa . Proteins corresponding in mass to the first four of these also occur in the mosquitocidal strain, B . thuringiensis subsp . israelensis (BTI), and it is thought therefore that the mosquitocidal activity of both strains is due to these four proteins . In other studies it has been shown that each of these proteins exhibits from moderate to high toxicity to mosquitoes, though the specific toxicity of the 144 kDa protein in PG-14 to mosquitoes remains unknown . In the present study, two parasporal body mutants (M146 and M242) of PG-14 were developed growing the wild-type strain at 42 degrees C . The parasporal body of M146 contained less of the 65-kDa protein and was less toxic (LC50 = 108 ng/ml) to mosquitoes than the wild-type strain (LC50 = 8.3 ng/ml) . The parasporal body of M242 consisted of a bipyramidal crystal composed of a 144-kDa protein that was not toxic to the mosquito, Aedes aegypti, but exhibited substantial toxicity (LC50 = 2.5 micrograms/ml) to the lepidopteran . Trichoplusia ni . Because the parasporal bodies of BTI and BTM PG-14 are similar in mosquitocidal toxicity on a weight basis, the latter results suggest the 144-kDa protein, though not mosquitocidal alone, can contribute to mosquitocidal, activity when in the presence of other mosquitocidal proteins.

J Bacteriol, 1990 Jan, 172(1), 481 - 3
Characterization of the parasporal inclusion of Bacillus thuringiensis subsp . kyushuensis; Held GA et al.; Electron microscopy of Bacillus thuringiensis subsp . kyushuensis revealed that the parasporal inclusions are composed of a homogeneous center surrounded by a thick, electron-dense coating . Antibodies directed against the 135- and 65-kilodalton B . thuringiensis subsp . israelensis peptides cross-reacted with the 70- and 26-kilodalton peptides, respectively, of B . thuringiensis subsp . kyushuensis.

DNA Seq, 1990, 1(2), 97 - 106
Models for the structure and function of the Bacillus thuringiensis delta-endotoxins determined by compilational analysis; Hodgman TC et al.; The protein delta-endotoxins of Bacillus thuringiensis are a commercially and environmentally important class of highly specific insecticides . From an alignment of their sequences, certain structural and functional domains can be inferred which may shed light on the mode of action of these toxins.

Cancer Immunol Immunother, 1990, 32(2), 95 - 104
Major histocompatibility complex class II antigen expression during potentiation of line-10 tumor immunity after intralesional administration of bacillus Calmette-Guérin; Steerenberg PA et al.; Intralesional injection of BCG into an established line-10 hepatocellular carcinoma in the strain-2 guinea pig causes regression of the tumor and induction of line-10 immunity . We found that the animals were already protected for a second challenge with line-10 tumor cells 7 days after BCG treatment . We studied whether this early induction of immunity was correlated with the expression of MHC class II antigens on line-10 tumor cells and was correlated with an increased expression of MHC class II antigens on leukocytes in the primary tumor and in the regional lymph node (Ln . axillaris accessorius) . The MHC class II antigens and the leukocyte subpopulations were measured with monoclonal antibodies and flow cytofluorometry . In the draining lymph node the number of nucleated cells increased about 10-fold during the first 5 days after intralesional injection of BCG . At this time the MHC class II antigen expression of these cells was increased from 21%-32% in the naive controls to 39%-53% in animals with BCG-treated tumors . This implies that the number of MHC-class-II-positive cells increased about 20-fold in the draining lymph node . Surprisingly, the increase in percentage of MHC-class-II-antigen-positive cells was mainly due to an increase of IgM-positive B cells from 8%-11% to 22%-41% and an increase of IgG-positive B cells from 7%-27% to 25%-44% . In the tumor, BCG treatment induced a small increase of MHC-class-II-antigen-positive cells from 11%-12% to 15%-20% . Probably this increase came not from tumor cells but mainly from a BCG-induced infiltration of mononuclear cells, as an increase of T cells from 14% to 20%, an increase of macrophages from 8% to 18%, and an increase of B cells from 0 to 6% was observed . We conclude that the potentiation of anti-(line-10 tumor cell) immunity correlated with a 20-fold increase of MHC-class-II-antigen-positive cells in the lymph nodes and a small increase in the number of MHC-class-II-antigen-positive tumor-infiltrating cells.

Jpn J Ophthalmol, 1990, 34(3), 306 - 13
Immunosuppressive effect of gramicidin S on experimental ocular neuritis and allergic encephalomyelitis; Matsushima S et al.; Investigations were carried out on the immunosuppressive effect of gramicidin S (GrS), a cyclic peptide antibiotic produced by Bacillus brevis, on the onset of experimental ocular neuritis and allergic encephalomyelitis in Lewis rats immunized with rat brain homogenates . The criteria for evaluation of the drug effect were changes in body weight, activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase), clinical manifestations such as paralysis of lower extremities and histopathological changes . Clinical symptoms and body weight reduction were effectively prevented by GrS treatment of immunized animals . The activity of the myelin marker enzyme CNPase was markedly decreased in the lumbar spinal cord of encephalitogen-immunized animals on day 16 (ie 16 days after immunization) and the decrease of enzymatic activity was partially prevented by GrS administration . On the other hand, the CNPase activity of the retrobulbar optic nerve of inoculated animals remained essentially the same as that of healthy control animals, although inflammatory changes were prominent in the optic nerve . Histopathological changes observed in the optic nerve and spinal cord of diseased animals were virtually absent in GrS-treated animals . A possible mechanism of the immunosuppressive activity of GrS is discussed.

Crit Rev Biotechnol, 1990, 10(3), 223 - 40
Molecular biology of antibiotic production in Bacillus; Nakano MM et al.; Several species of the genus Bacillus produce peptide antibiotics which are synthesized either through a ribosomal or non-ribosomal mechanism . The antibiotics gramicidin, tyrocidine, and bacitracin are synthesized nonribosomally by the multienzyme thiotemplate mechanism . Surfactin and mycobacillin are also synthesized nonribosomally but by a mechanism that, apparently, is distinct from that of the multienzyme thiotemplate . Other antibiotics such as subtilin are gene encoded and are ribosomally synthesized . Molecular genetic and DNA sequence analysis have shown that biosynthesis genes for some antibiotics are clustered into polycistronic transcription units and are under the control of global regulatory systems that govern the expression of genes that are induced when Bacillus cells enter stationary phase of growth . Future experiments involving the molecular dissection of peptide antibiotic biosynthesis genes in Bacillus will be attempted in hopes of further examining the mechanism and regulation of antibiotic production.

Nauchnye Doki Vyss Shkoly Biol Nauki, 1990, (3), 48 - 55
{The radiobiological characteristics of the P+ variant of Bacillus brevis var . G.--B . in relation to gramicidin S synthesis}; Parkhomenko IM et al.; The radiosensitivity of P(+) variant Bacillus brevis var . G.-B . cells cultured under condition of normal and inhibited gramicidin S synthesis, antibiotically high-active strain and high radioresistant cells has been studied . It has been shown that the radioresistance of bacterial cells correlates, in general, with their antibiotic activity: the antibiotic superproduced is more radioresistant than P(+) variant, the inhibition of antibiotic synthesis by beta-phenil-beta-alanin rises a little the sensitivity of P(+) variant cells . But the radioresistant fraction of P(+) variant contains the lower antibiotic amount than the whole population . It has been concluded that the radioprotective action of gramicidin S can not be the only reason of the above-mentioned differences in radiosensitivity.

Eur Urol, 1990, 17(4), 338 - 42
Immunohistochemical markers defined by monoclonal antibodies and response to bacillus Calmette-Guérin endovesical immunotherapy for superficial bladder tumors; Flam TA et al.; As bacillus Calmette-Guerin (BCG) immunotherapy is highly effective for most but not all superficial bladder tumors, there is a need to define predictors of response to this mode of treatment . We have investigated a panel of markers defined by monoclonal antibodies, directed against tumor-associated transitional cell carcinoma antigen (G4 and E7), epidermal growth factor receptor, cytokeratin (CK) 18 and blood group antigens A, B and H, using an indirect immunoperoxidase staining on paraffin sections . Twenty superficial bladder tumors (T1) treated with intravesical BCG therapy (10 responders and 10 nonresponders) were tested with this panel . Among the responders, expression of CK18 antigen was positive in 7 and negative in 3, whereas in the nonresponder group it was positive in 2 and negative in 8 . The difference was statistically significant (p less than 0.05) . Loss of expression of CK18 antigenicity was associated with recurrence or progression of superficial bladder tumors following BCG therapy, indicating that changes in CK patterns should be investigated as potential predictive markers for response to BCG.

Prikl Biokhim Mikrobiol, 1990 Jan-Feb, 26(1), 85 - 92
{Antigenic structure of endotoxins from Bacillus thuringiensis}; Iagudin VIa et al.; The proteins of parasporal crystals of 9 strains of entomopathogenic bacteria of the Bacillus thuringiensis group of different H-serotypes were studied by different methods . Electrophoresis showed that crystals from the bacteria of 1, 3, 4, 5, 8, and 10 serotypes contained 1-2 proteins with a molecular weight of 130-145 kD and an additional fraction with a molecular weight of 55-120 kD . Double-radial immunodiffusion revealed that the crystals of all the serotypes studied are immunologically related . The proteins contained at least 3 types of antigens that can be detected either separately or all together when being tested with different antisera . The ELISA technique showed that the crystal proteins homologous with respect of antigens gave, as a rule, similar titration results.

Prikl Biokhim Mikrobiol, 1990 Jan-Feb, 26(1), 50 - 4
{Various physico-chemical properties of the amylolytic complex from Bacillus mesentericus}; Koltukova NV et al.; Some physico-chemical properties of the Bacillus mesentericus amylolytic complex were studied, and optimal conditions of starch hydrolysis (pH 7.5-8.0; 45 degrees C) were found . The half-life of amylases at 50 degrees was 75 min . The heat stability of the enzymes increased in the presence of Ca2+ ions . Amylase was stable at pH 7-9 and readily inactivated at pH below 6.0 . By physical and chemical characteristics the complex was close to analogous preparations from Bacillus alkalophilic strains . Isoelectrofocusing revealed that the complex consisted at least of two amylolytic enzymes.

Agric Biol Chem, 1990 Jan, 54(1), 197 - 201
Comparison of CD composition produced by chimeric CGTases; Kaneko T et al.; Using 12 chimeric cyclomaltodextrin glucanotransferases (CGTases) constructed from two genes, the CGTase gene from the alkalophilic Bacillus sp . strain No . 38-2 (CGTase 38-2 gene) and the CGTase gene from the alkalophilic Bacillus sp . strain No . 17-1 (CGTase 17-1 gene), we compared the effect of those chimeric enzymes on cyclodextrin (CD) production, especially on the composition of the CDs produced . It was found that the N-terminal and the C-terminal segments were important for CD production . Chimeric enzymes that contained the N-terminal and the C-terminal segments derived from CGTase 38-2 produced large amounts of CD, and especially a higher proportion of alpha-CD than those of other chimeric enzymes.

Biochemistry, 1989 Dec 26, 28(26), 9908 - 12
Common structural changes accompany the functional inactivation of HPr by seryl phosphorylation or by serine to aspartate substitution; Wittekind M et al.; Although many proteins are known to be regulated via reversible phosphorylation, little is known about the mechanism by which the covalent modification of seryl, threonyl, or tyrosyl residues alters the activities of the target systems . To address this question, modified versions of Bacillus subtilus HPr, a protein component of the bacterial phosphotransferase system, have been studied by 1H NMR spectroscopy . Phosphorylation at Ser46 or a Ser to Asp substitution at this position inactivates HPr {Reizer, J., Sutrina, S . L., Saier, M . H., Stewart, G . C., Peterkofsky, A., & Reddy, P . (1989) EMBO J . 8, 2111-2120} . Two-dimensional spectra of these two modified proteins display nearly identical proton chemical shifts that differ significantly from those observed in the spectra of the unphosphorylated, wild-type protein and of functionally active HPr mutants . The results demonstrate that the functional inactivation of HPr brought about by the serine to aspartate mutation is accompanied by the same structural changes that occur when HPr is phosphorylated at Ser46.

Cancer Res, 1989 Dec 15, 49(24 Pt 1), 7045 - 50
Characterization of IgG and IgM antibodies induced in melanoma patients by immunization with purified GM2 ganglioside; Livingston PO et al.; The ganglioside GM2 is a differentiation antigen expressed on the cell surface of human malignant melanomas and other cancers of neuroectodermal origin . We have previously reported that immunization with purified GM2 combined with Bacillus Calmette-Guerin as adjuvant and pretreatment with low-dose cyclophosphamide induced production of antibodies against GM2 in five of six patients . We have now extended our study and analyzed the induced antibodies against GM2 were not detected . After immunization, high-titer IgM antibodies were induced in 17 of 24 patients, and high-titer IgG antibodies in eight cases . Additional treatment of 12 patients with cimetidine, a histamine H2 receptor antagonist reported to have antisuppressor cell activity, had no effect on GM2 antibody titers . Antibodies against asialo-GM2 were present in all patients, and antibodies against GM1 were present in 33% of patients, before and after immunization . Antibodies induced by immunization were specific for GM2, though some reacted predominantly with N-acetylneuraminic acid GM2 (GM2), and some reacted with GM2 and N-glycolylneuraminic acid GM2(NeuGcGM2) . The pattern of reactivity with GM2 is consistent with the response to T-cell-independent antigens: both IgM and IgG antibody responses against GM2 were short lived; peak titers seen after initial and secondary vaccinations were similar; and delayed-type hypersensitivity responses to skin test challenges with GM2 were not detected in any patients . However, the IgG response typed as predominantly IgG1 and IgG3, not IgG2 as might be expected for carbohydrate antigens (which are generally T-cell-independent antigens) . Because IgG1 and IgG3 antibody responses are usually to T-cell-dependent antigens, the humoral immune response elicited by GM2 vaccination has both T-cell-dependent and T-cell-independent characteristics . These IgM and IgG responses against this neuroectodermal differentiation antigen expressed on melanoma cells have been induced without evidence of neurological or other toxicity.

Eur J Biochem, 1989 Dec 8, 186(1-2), 239 - 47
Specificity of Bacillus thuringiensis delta-endotoxins . Importance of specific receptors on the brush border membrane of the mid-gut of target insects; Van Rie J et al.; To study the molecular basis of differences in the insecticidal spectrum of Bacillus thuringienesis delta-endotoxins, we have performed binding studies with three delta-endotoxins on membrane preparations from larval insect mid-gut . Conditions for a standard binding assay were established through a detailed study of the binding of 125I-labeled Bt2 toxin, a recombinant B . thuringiensis delta-endotoxin, to brush border membrane vesicles of Manduca sexta . The toxins tested (Bt2, Bt3 and Bt73 toxins) are about equally toxic to M . sexta but differ in their toxicity against Heliothis virescens . Equilibrium binding studies revealed saturable, high-affinity binding sites on brush border membrane vesicles of M . sexta and H . virescens . While the affinity of the three toxins was not significantly different on H . virescens vesicles, marked differences in binding site concentration were measured which reflected the differences in in vivo toxicity . Competition experiments revealed heterogeneity in binding sites . For H . virescens, a three-site model was proposed . In M . sexta, one population of binding sites is shared by all three toxins, while another is only recognized by Bt3 toxin . Several other toxins, non-toxic or much less toxic to M . sexta than Bt2 toxin, did not or only marginally displace binding of 125I-labeled Bt2 toxin in this insect . No saturable binding of this toxin was observed to membrane preparations from tissues of several non-susceptible organisms . Together, these data provide new evidence that binding to a specific receptor on the membrane of gut epithelial cells is an important determinant with respect to differences in insecticidal spectrum of B . thuringiensis insecticidal crystal proteins.

Eur J Biochem, 1989 Dec 8, 186(1-2), 389 - 93
Enzymatic properties of isozymes and variants of glucose dehydrogenase from Bacillus megaterium; Mitamura T et al.; Three glucose dehydrogenases (GlcDH) from Bacillus megaterium, GlcDH-I, GlcDH-II and GlcDH-IWG3, were purified from Escherichia coli cells harboring one of the hybrid plasmids, pGDK1, pGDK2 and pGDA3, respectively, pGDK1 and pGDK2 contain two isozyme genes, gdhI and gdhII, respectively, from B . megaterium IAM 1030 and pGDA3 contains an isozyme gene from B . megaterium IWG3; GlcDH-IWG3 is a variant of GlcDH-I . GlcDH-I and GlcDH-II have similar pH/activity profiles and the profile for GlcDH-IWG3 is identical to that of GlcDH-I . The pH/stability profiles of these enzymes show that GlcDH-IWG3 is the most stable enzyme in the acidic region, while GlcDH-II is the most stable in the alkaline region, and GlcDH-I is the most unstable throughout the entire pH range examined . As for thermostability, GlcDH-II is the most resistant against heat inactivation at pH 6.5 . The values of the first-order rate constant for heat inactivation at 50 degrees C are 0.27 min-1, 0.05 min-1 and 0.11 min-1 for GlcDH-I, GlcDH-II and GlcDH-IWG3, respectively . Kinetic studies show that these enzymes have similar kinetic constant values except that there are some differences in Kia for NAD(P) and Ka (the limiting Michaelis constant) for NAD; the values of the ratio of Kia for NAD and NADP are 11,340 and 8.7 for GlcDH-I, GlcDH-II and GlcDH-IWG3, respectively . GlcDH-I and GlcDH-IWG3 have very similar substrate specificities and GlcDH-II has a slightly higher specificity for D-glucose and 2-deoxy-D-glucose than the others . The results are discussed on the basis of the amino acid substitutions between the enzymes.

Int J Lepr Other Mycobact Dis, 1989 Dec, 57(4), 801 - 9
Skin test studies on close contacts of leprosy patients in India; Lord R et al.; Skin-test studies with a series of tuberculins have been carried out in close contacts of multibacillary (MB) leprosy patients around three leprosy centers in India, and casual contacts of the disease around two centers . The results show that the rate of acquisition of leprosin A positivity is associated with age and the closeness of contact with MB leprosy . At the age of 15 years, the differences between the two types of contact were highly significant (p less than 0.00001) . Many responses to leprosin A are directed toward the group iv species-specific, antigens of the leprosy bacillus, and the significance of positivity is discussed in relation to protective immunity from leprosy . The differences from Iran show that positivity to leprosin A is not solely the effect of the degree of contact with the disease, but must also have a genetic or environmental element, the latter being favored . The results from Miraj show that the high levels of tuberculin, scrofulin, and vaccin positivity seen in Fathimanagar, and to a lesser extent in Karigiri, are not a consequence of contact with leprosy . BCG vaccination made little difference to the leprosin A positivity of close contacts of leprosy patients, although it significantly enhanced positivity among casual contacts around Miraj (p less than 0.002) . BCG vaccination significantly increased tuberculin positivity in Miraj and Karigri, and in those under 11 years of age in Fathimanagar . It made no difference to the already high level of positivity found in older persons around Fathimanagar.(ABSTRACT TRUNCATED AT 250 WORDS)

Semin Respir Infect, 1989 Dec, 4(4), 318 - 25
Prevention of tuberculosis; Starke JR; Tuberculosis (TB) continues to be a major cause of morbidity and mortality throughout the world . One billion people are infected with the tubercle bacillus, and there are 8 million new cases and 3 million deaths annually . Existing methods of preventing TB are effective, but due to economic and societal constraints, have not been applied throughout the world . Examples of primary prevention include case management and contact investigation, environmental control and vaccination . Secondary prevention is achieved by chemotherapy . The efficiency and use of these various methods will be reviewed, and areas for future study will be discussed.

Appl Environ Microbiol, 1989 Dec, 55(12), 3178 - 83
Two Bacillus beta-mannanases having different COOH termini are produced in Escherichia coli carrying pMAH5; Akino T et al.; The nucleotide sequence was determined for the alkalophilic Bacillus sp . strain AM-001 beta-mannanase gene which produced two beta-mannanases (A and B) in Escherichia coli transformants . The putative beta-mannanase gene was 1,539 base pairs long and encoded a mature beta-mannanase protein of 487 amino acids and a signal peptide of 26 amino acids . The COOH-terminal amino acid of beta-mannanase A is an arginine residue located at amino acid 513 of the deduced amino acid sequence, and that of beta-mannanase B is a valine residue located at amino acid 365 . Deletion derivatives having 1,098 base pairs from the ATG start codon maintained the beta-mannanase activity of the encoded polypeptide . However, clones harboring DNA fragments (1,051 base pairs) shorter than the gene which encoded beta-mannanase B (1,095 base pairs) did not exhibit the beta-mannanase activity . The simultaneous production of both beta-mannanases A and B in an E . coli transformant was demonstrated by the maxicell procedure.

Zhonghua Jie He He Hu Xi Za Zhi, 1989 Dec, 12(6), 336 - 7, 380-1
{Tuberculosis of the breast}; Gong TQ; Tuberculosis of the breast is a rare malady . The clinician may confuse tuberculosis of the breast with either carcinoma or breast abscess . In the past 30 years at the First Teaching Hospital attached to Xi'an Medical University, only 23 patients reported herein had documented tuberculosis of the breast . Tuberculosis of the breast is a disease of younger women between 20 and 40 years of age . Mammary tuberculosis may be primary or secondary . There are three recognized modes of spread of the tubercle bacillus to the breast: hematogenous, lymphatic spread, and direct extension . There are three recognized types of mammary tuberculosis: nodular, diffuse, and sclerosing type . The diagnosis of mammary tuberculosis is difficult . The most reliable diagnostic studies include bacteriologic cultures of aspirate, histologic examination of tissue, and guinea pig inoculation . The differential diagnosis includes with carcinoma, acute or chronic mastitis with abscess . The treatment of tuberculous mastitis requires a combination of surgery and antituberculous drugs.

J Biochem (Tokyo), 1989 Dec, 106(6), 966 - 71
Isolation and fundamental properties of a phospholipase A2 inhibitor from the blood plasma of Trimeresurus flavoviridis; Kogaki H et al.; Phospholipase A2 inhibitor was purified from the blood plasma of Habu, Trimeresurus flavoviridis, by Sephadex G-200 gel filtration, DEAE-cellulose chromatography, and Blue-Sepharose CL-6B column chromatography . The purified inhibitor was shown to be a glycoprotein with a molecular weight of about 100K . It was found to consist of four subunits whose molecular weights were around 20-24K . In order to examine the inhibition mechanism of the inhibitor, the interaction of the inhibitor with a phospholipase A2 from T . flavoviridis venom was examined by Sephadex G-100 gel filtration . One inhibitor molecule was found to bind directly to one phospholipase A2 molecule in both the presence and absence of Ca2+ . The inhibitor inhibited the phospholipase A2 from T . flavoviridis venom with an apparent dissociation constant, Ki, of 1.7 X 10(-10) M, but not the porcine pancreas enzyme or the Agkistrodon halys blomhoffii enzyme belonging to the same family, Crotalidae, as T . flavoviridis, or the phospholipase C from Bacillus cereus.

Minerva Ginecol, 1989 Dec, 41(12), 609 - 14
{Activity of topical flunoxaprofen in nonspecific vaginitis . Comparison with meclocycline sulfosalicylate}; Facchini V et al.; A trial was performed in 30 patients affected by non-specific vaginitis . The results show that the topical application (by vaginal washings) of flunoxaprofen produces a high therapeutic activity like that of meclocycline . Contrary to meclocycline, flunoxaprofen does not possess bactericidal and bacteriostatic properties . Therefore, the normalization of vaginal flora, with a remarkable increase of Doderlein bacillus, is exclusively due to local antiphlogistic activity of flunoxaprofen . Contrary to meclocycline, flunoxaprofen induces a precocious increase of Doderlein bacillus, necessary for a definitive normalization and to limit the possibility of relapses.

Acta Neurol (Napoli), 1989 Dec, 11(6), 390 - 9
Cerebral listeriosis in adults . Three cases report; Masini T et al.; The Authors describe three autoptic cases of meningoencephalitis in adults due to an emerging pathogen, the Gram-positive bacillus Listeria monocytogenes . Only one of these was an immunosuppressed patient (recurrent carcinoma of the larynx and bladder papillomas), while no severe cause of debilitation was detected in the other two cases, excluding colonic diverticulosis in both . The post-mortem examination revealed leptomeningitis and encephalitis . Lympho-monocytic tissue reaction with perivascular cuffing was present in all the cases . The etiologic agent, L . monocytogenes, was identified by cultures from clinical or autoptic specimens.

Kekkaku, 1989 Dec, 64(12), 806 - 10
{A study on primarily treated tuberculosis cases with positive sputum}; Naoe H; A total of 377 cases with primarily treated bacillary tuberculosis selected from 432 patients admitted to 5 major national sanatoria during 1987 was analysed and compared with the same sort of studies done in national sanatoria in 1976 and 1980, and in addition 21 dead cases were investigated . The results were as follows . 1 . 110 cases (29%) were over 60 years of age . 2 . New intensive regimens containing INH and RFP has become popular (over about 70%) . 3 . The duration of chemotherapy has been shortened (65% terminated within 12 month, while only 9.5% in 1980) . 4 . The duration of admission has been shortened (65% discharged within 6 month, while 59% 1980) . 5 . 21 dead cases were in the higher age group (70% were over 60 years of age) and the higher rate of complications including diabetes mellitus (28%), cancer (21%) and heart failure (19%) . 6 . At the start of chemotherapy, chest X-ray showed fresh types of GAKKEN A & B in 307 cases (82%) and cavitary type of GAKKAI I & II in 274 cases (73%) and these proportions have not changed since 1980 and the response to chemotherapy was as good as in 1980 . Based on the above findings, the older age patients with various complications might be regarded as the most difficult cases to be cured at present in coming years.

Urology, 1989 Dec, 34(6), 381 - 2
BCG in management of transitional cell carcinoma invasive to prostate; Siami P et al.; Transitional cell carcinoma invading the prostate is a difficult problem to manage . Transurethral resection of the bladder neck followed by intravesical bacillus Calmette-Guerin (BCG) is one suggested option.

J Bacteriol, 1989 Dec, 171(12), 6656 - 67
Characterization of a dynamic S layer on Bacillus thuringiensis; Luckevich MD et al.; The surfaces of three Bacillus thuringiensis strains possess an S layer composed of linear arrays of small particles arranged with p2 symmetry and with a = 8.5 nm, b = 7.2 nm, and gamma = 73 degrees . Platinum shadows of whole cells and S-layer fragments revealed the outer surface of the array to be smooth and the inner surface to be corrugated . Treatment with 2 M guanidine hydrochloride at pH 2.5 to 4 best removed the S layer for chemical characterization; it was a relatively hydrophilic 91.4-kilodalton protein with a pI of 5, no detectable carbohydrate, cysteine, methionine or tryptophan, and 21.2% nonpolar residues . No N-terminal homology with other S-layer proteins was evident . Antibody labeling experiments confirmed that the amount of S layer was proportional to the growth phase in broth cultures . Late-exponential- and stationary-growth-phase cells typically sloughed off fragments of S layer, and this may be the result of wall turnover . Indigenous autolytic activity in isolated walls rapidly digested the wall fabric, liberating soluble S-layer protein . At the same time, proteases frequently reduced the molecular weight of the 91.4-kilodalton protein, but these polypeptides could still be identified as S-layer components by immunoblotting . As cultures were serially subcultured, the frequency of appearance of the S layer diminished, and it was eventually lost . The dynamic nature of this S layer makes it atypical of most previously identified S layers and made it unusually difficult to characterize.

J Bacteriol, 1989 Dec, 171(12), 6637 - 48
Cloning of the crystalline cell wall protein gene of Bacillus licheniformis NM 105; Tang M et al.; A protein with a tetragonal pattern, defined as RS protein, was found on the wall surface of an alkaline phosphatase secretion-deficient mutant (NM 105) of Bacillus licheniformis 749/C . The protein was present on the wall surface of the exponential-growth-phase cells, but at the stationary growth phase it was overproduced and hypersecreted . This protein was precipitated to homogeneity from the culture fluid by 80% ammonium sulfate saturation and chilled acetone . The molecular mass of the protein was 98 kilodaltons, and it had a single subunit in a sodium dodecyl sulfate gel . Specific anti-RS antibody was generated in rabbits and used to immunolabel the RS protein on the cells at different growth phases . In early-exponential-growth-phase cells, the outside surface of the wall, the cytoplasm, and the inside surface of the cytoplasmic membrane were labeled . In stationary-growth-phase cells, the cytoplasm was poorly labeled, but the labeling on the outside surface of the wall was high . AB . licheniformis NM 105 gene library was made by using the lambda phage EMBL3 . The RS protein expression from this gene library was detected by a modified autoradiographic procedure . One of the amplified RS protein-positive plaques (4213-1) containing recombinant DNA was chosen, and the restriction map of this DNA was prepared . The RS protein expressed in Escherichia coli NM 539 infected with 4213-1 recombinant phage had a lower molecular mass than the purified authentic RS protein . The 4.5-kilobase-pair (kbp) SalI-EcoRI fragment of the recombinant DNA was cloned in the shuttle plasmid pMK4 to construct pMK462, which was expressed in B . subtilis MI112 and produced the RS protein identical in molecular mass to the purified authentic RS protein . The RS protein expression was also demonstrated in cryosections of transformed E . coli and B . subtilis cells by immunoelectron microscopy . The 1.2-kbp SalI-HindIII and 1.8-kbp HindIII-HindIII recombinant DNA restriction enzyme fragments, respectively, from the right of the restriction map produced anti-RS antibody cross-reacting proteins . The expression of the 1.2-kbp SalI-HindIII DNA fragment cloned in pUC8 could be induced with isopropyl-beta-D-thiogalactopyranoside . The 1.8-kbp DNA restriction fragment hybridized with both the chromosomal DNA of strain NM 105 and the recombinant phage 4213-1 DNA . The RS gene expression was finally demonstrated in transformed E . coli 539 cells by in situ hybridization of frozen thin sections with the 1.8-kbp HindIII biotin-dATP probe and immunolabeling these with anti-biotin immunoglobulin G and protein A-gold.

J Vasc Surg, 1989 Dec, 10(6), 688 - 92
Tuberculous aneurysm of the femoral artery: a complication of bacille Calmette-Guérin vaccine immunotherapy--a case report; Bornet P et al.; A case of bilateral femoral mycotic aneurysms resulting from bacille Calmette-Guerin vaccine, is reported in a 74-year-old man treated by intravesical immunotherapy . The diagnosis was made after histologic and bacteriologic examinations and biochemical analysis of the acid-fast-baccilli . Treatment consisted of resection and replacement of the femoral arteries, with an expanded polytetrafluoroethylene graft on the left side and a saphenous vein graft on the right side . After surgery the patient was treated with antituberculous chemotherapy for 1 year . The patient was doing well 18 months later.

J Urol, 1989 Dec, 142(6), 1438 - 41
Choice of an optimal diluent for intravesical bacillus Calmette-Guerin administration; Hudson MA et al.; The physical conditions, including diluent pH, salt concentration and duration of bacillus Calmette-Guerin attachment, were determined in in vitro binding assays for soluble and matrix fibronectin . Since soluble fibronectin may block attachment of bacillus Calmette-Guerin to matrix fibronectin in the bladder, the optimal conditions were determined under which matrix fibronectin-bacillus Calmette-Guerin binding was maximal and soluble fibronectin-bacillus Calmette-Guerin binding was minimal . These conditions, which were confirmed in vivo in the murine bladder model, included use of normal saline, pH 7 as diluent for bacillus Calmette-Guerin organisms, with retention of the bacillus Calmette-Guerin suspension for 2 hours.

J Immunol, 1989 Dec 1, 143(11), 3517 - 23
IL-6 inhibits lipopolysaccharide-induced tumor necrosis factor production in cultured human monocytes, U937 cells, and in mice; Aderka D et al.; Incubation of the human U937 histiocytic lymphoma cell line with granulocyte-macrophage colony stimulating factor (GM-CSF) rendered the cells responsive to induction of TNF by LPS . Treatment with IL-6 reduced TNF production in GM-CSF-primed U937 cells . The inhibitory effect was most pronounced (approximately equal to 80%) when IL-6 was added either along with GM-CSF or within the first 3 h of GM-CSF treatment . Both GM-CSF or IL-6 inhibited {3H}TdR uptake in U937 cells, and simultaneous treatment with GM-CSF and IL-6 resulted in an additive inhibitory effect on cell proliferation . However, the inhibition of TNF production could not be explained by the inhibitory effect of IL-6 on cell growth, nor was it due to a reduction in cell viability . An inhibition of TNF production by IL-6 was also demonstrated in cultured human peripheral blood monocytes . Treatment with IL-6 also resulted in a dose-dependent reduction of the 17-kDa TNF band revealed by SDS-PAGE after labeling monocytes with {35S}cysteine and immunoprecipitation with anti-TNF mAb . In addition, treatment with IL-6 resulted in a reduction of monocyte in vitro cytotoxicity for tumor target cells . Finally, in mice sensitized by the administration of Bacillus Calmette-Guerin, the injection of IL-6 significantly reduced the levels of TNF found in the serum upon challenge with LPS . Inasmuch as TNF is known to be an inducer of IL-6, the inhibitory action of IL-6 on TNF production may represent the negative arm of a regulatory circuit . The inhibitory action of IL-6 on TNF production is consistent with a predominantly antiinflammatory role of IL-6 in the intact organism.

Hansenol Int, 1989 Dec, 14(2), 107 - 11
{Efficacy of multidrug therapy in treatment of paucibacillary leprosy patients--preliminary results}; Avelleira JC et al.; The efficaciousness evaluation of the polychemotherapics regimens recommended by WHO to the hansenian paucibacillary patients, is carried out mainly by the suitable follow up of patients after therapeutic discharge . The criterion for inclusion of patients as paucibacillary ones is another point of importance . The authors based on the follow up of 66 patients that completed treatment and in the absence of relapses until the moment, advise that together with the clinical classification it should be considered factors like: the Mitsuda test, the number of lesions and the bacilloscopy result.

Wei Sheng Wu Xue Bao, 1989 Dec, 29(6), 397 - 404
{Proteins and proteinase-resistant polypeptides of delta-endotoxins from Bacillus thuringiensis and their insecticidal activity}; Li RS et al.; Proteins, solubility in alkaline buffer and proteinase-resistant polypeptides (PRP) of parasporal crystals (delta-endotoxin) from Bacillus thuringiensis were investigated . 19 sorts of crystals were divided into 7 types . Difference of crystalline proteins were coincident with their solubility and PRP . The characters of the protein, solubility and PRP were subspecies-specific basically, but in a few of crystals they were strain-specific . There was a closed relation between the biochemistry property of crystal protein and the toxicity in 2 species of insects . Crystal with protein of MW 130-138 kD kD or MW 130-138 kD and 60-65 kD, and PRP of MW 68-75 kD were high toxic in Bombyx mori (Lepidoptera), but most of them only showed weak or no toxicity in Culex fatigans (Diptera) . Crystals containing more than 3 sorts of proteins with MW 15-138 kD in major protein and MW 35-75 kD in minor protein and PRP of 35-65 kD had very strong toxicity in Culex fatigans, but no toxicity in Bombyx mori . The construct of the crystal protein and the importance of the character in protein and PRP for the selection of expected strain and for the work of genetic engineering in Bacillus thuringiensis were discussed.

Am J Clin Pathol, 1989 Dec, 92(6), 813 - 20
Malacoplakia of the thyroid gland; Katoh R et al.; A case of malacoplakia of the thyroid gland is described in a 50-year-old Japanese woman . This lesion clinically mimicked a malignant neoplasm, and the true diagnosis of malacoplakia was made only after histologic examination; light microscopy revealed a granulomatous nodule with an accumulation of von Hansemann's histiocytes containing PAS-positive and von Kossa's-positive intracytoplasmic and extracytoplasmic inclusions known as Michaelis-Gutmann bodies . There were some foci consisting of neoplasm-like or hyperplastic residual follicles within the lesion . Electron microscopically, a small number of bacilliform organisms were demonstrated within the lesion . X-ray microanalysis of Michaelis-Gutmann bodies was performed and revealed the presence of phosphorus, calcium, iron, and chloride . It is suggested that the malacoplakic lesion may be associated with the hyperplastic or neoplastic follicular lesion, and bacterial infection could be important in the causation of malacoplakia of the thyroid gland.

J Gen Microbiol, 1989 Dec, 135 ( Pt 12), 3447 - 57
Construction of a chimeric series of Bacillus cyclomaltodextrin glucanotransferases and analysis of the thermal stabilities and pH optima of the enzymes; Kaneko T et al.; The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from the alkalophilic Bacillus sp . strain no . 17-1 was cloned in Escherichia coli . The cloned CGTase gene consisted of a single open reading frame which would encode a polypeptide of 713 amino acids, and the first 27 amino acid residues comprised a signal peptide . The nucleotide sequence and the amino acid sequence of this CGTase (CGTase 17-1) gene had strong homology with those of the CGTase (CGTase 38-2) gene previously cloned in our laboratory from the alkalophilic Bacillus sp . strain no . 38-2, although the enzymic properties of the CGTase 17-1 were distinct from those of the CGTase 38-2 . To analyse those enzymic properties further, we constructed 12 chimeric CGTases using three restriction nuclease sites and compared the enzymic properties of the chimeric CGTases . The N-terminal part of the enzyme was important for heat stability, and the pH-activity profile was influenced by both the N- and the C-terminal parts . A third segment was less important for enzymic properties.

Gifu Shika Gakkai Zasshi, 1989 Dec, 16(2), 441 - 63
{Biochemical studies on intramembranous localization of alkaline phosphatase in Bacillus megaterium KM}; Takahashi K; Intramembranous localization of alkaline phosphatase (orthophosphoric monoester phosphohydrolase, alkaline optimum, EC 3, 1, 3, 1; AlPase) was observed biochemically in Bacillus megaterium KM grown in 1% polypeptone medium containing 0.5% NaCl at 37 degrees C under aerobic conditions and harvested at the latter logarithmic phase . AlPases from B . megaterium have been separated into soluble and membrane-bound forms by the centrifugation after cell disruption by sonication . The membrane-bound enzyme was further fractionated to two forms by phase separation using a non-ionic detergent, Triton X-114; one was successfully solubilized into the aqueous phase and the other remained in the Triton phase . Both AlPases of sonication- and Triton-solubilized forms were partially purified by gel filtration and anion-exchange column chromatographies . Their molecular weights were different (52,000 for soluble and 66,000 for Triton-solubilized forms) and the Vmax of the sonication-solubilized enzyme (227 nmol/min/mg protein) was 11-fold higher than that of the Triton-solubilized one although similar Km values (1.7 and 2.3 mM) were observed . Optimum pH of these enzymes tended to shift to a neutral range during the purification steps . These results suggest the multiplicity of AlPase anchoring to the membranes; 1) sonication-solubilized form which may be buried within the membrane lipids by its hydrophobic peptide and solubilized by the cell disruption, 2) detergent-solubilized form which may be bound loosely to the membrane by its hydrophobic domain and solubilized due to the amphiphilicity of enzyme protein, and 3) insolubilized form which may be bound fast to the membrane by its strong hydrophobicity and also have the function of enzymatic ability.

Indian J Exp Biol, 1989 Dec, 27(12), 1060 - 3
Isolation, characterization and biological activities of a toxin from Bacillus megaterium (B-23); Bhattacharyya B et al.; Fungitoxic substance was isolated from the culture filtrate of B . megaterium (B-23) . Age of culture and pH of medium influence the fungitoxicity of its culture filtrate . Partially purified toxin was thermolabile, non-dialysable, ethyl acetate soluble, vanillin-sulphuric acid positive and effective within a range of pH 5-9 . It exhibited maximum UV absorption at 224 nm . Its melting point was 242 degrees C . The efficacy of this compound was tested on 4 jute parasites namely, C . corchori, C . gloeosporioides, M . roridum and A . citri, of which M . roridum and C . corchori were least and most sensitive to the toxin respectively.

Appl Environ Microbiol, 1989 Dec, 55(12), 3173 - 7
Cloning and characterization of two genes from Bacillus polymyxa expressing beta-glucosidase activity in Escherichia coli; Gonzalez-Candelas L et al.; DNA fragments from Bacillus polymyxa which encode beta-glucosidase activity were cloned in Escherichia coli by selection of yellow transformants able to hydrolyze the artificial chromogenic substrate p-nitrophenyl-beta-D-glucopyranoside . Restriction endonuclease maps and Southern analysis of the cloned fragments showed the existence of two different genes . Expression of either one of these genes allowed growth of E . coli in minimal medium with cellobiose as the only carbon source . One of the two enzymes was found in the periplasm of E . coli, hydrolyzed arylglucosides more actively than cellobiose, and rendered glucose as the only product upon cellobiose hydrolysis . The other enzyme was located in the cytoplasm, was more active toward cellobiose, and hydrolyzed this disaccharide, yielding glucose and another, unidentified compound, probably a phosphorylated sugar.

FEMS Microbiol Lett, 1989 Dec, 53(3), 241 - 6
Nucleotide sequence of the gene encoding the active-site serine beta-lactamase from Actinomadura R39; Houba S et al.; The gene encoding the extracellular, active-site serine beta-lactamase of Actinomadura R39, previously cloned into Streptomyces lividans, has the information for the synthesis of a 304 amino acid protein, the amino terminal region of which has the characteristic features of a signal peptide . The Actinomadura R39 beta-lactamase is another member of the class A beta-lactamases . In particular, it shows high homology with the beta-lactamase of Bacillus licheniformis.

Proc Natl Acad Sci U S A, 1989 Dec, 86(23), 9159 - 62
Antibacterial peptides from pig intestine: isolation of a mammalian cecropin; Lee JY et al.; Pig small intestine was used as starting material for a batchwise isolation of a peptide fraction enriched in antibacterial activities against Escherichia coli (anti-Ec factor) and against Bacillus megaterium (anti-Bm factor) . Separation and further purification were by different types of chromatography . Sequence analysis showed the anti-Bm factor to be apparently similar to vasoactive intestinal peptide . The anti-Ec factor was found to have a 31-residue sequence that was cecropin-like . It was named cecropin P1 and its structure was confirmed by solid-phase synthesis . Synthetic cecropin P1 with and without C-terminal amide was assayed on eight different bacteria . Mobility comparison between synthetic and natural cecropin P1 indicates that the natural peptide has a free C-terminal carboxyl group.

Infect Immun, 1989 Dec, 57(12), 3689 - 94
Analysis of human antibody epitopes on the 65-kilodalton protein of Mycobacterium leprae by using synthetic peptides; Meeker HC et al.; In order to study antibody reactivity to the Mycobacterium leprae 65-kilodalton (kDa) antigen, peptides representing overlapping sequences of the 65-kDa protein were synthesized, and a recombinant protein expression system for r65-kDa was constructed . Mouse monoclonal antibodies and leprosy patient seroreactivity to peptides and r65-kDa were tested by an enzyme-linked immunosorbent assay . All seven of the monoclonal antibodies used in this study reacted with their previously defined epitopes when tested against peptides . All monoclonal antibodies also reacted with r65-kDa . Leprosy patient seroreactivity to peptides and r65-kDa was seen in about one-third of active multibacillary cases . Specimens from patients positive for antibodies to peptides were seen to recognize different epitopes than did mouse monoclonal antibodies used in this study . It is concluded that substantial differences exist between mouse monoclonal antibodies and human leprosy patient reactivity to the 65-kDa antigen and that human seroreactivity to the 65-kDa antigen is indicative of a highly elevated bacillary load.

Am J Med, 1989 Nov 30, 87(5A), 248S - 249S
Ciprofloxacin in the treatment of gram-negative bacillary meningitis; Schonwald S et al.; Gram-negative bacillary meningitis (typically caused by E . coli, K . pneumoniae, P . aeruginosa, A . calcoaceticus, and others) has been associated with head trauma, neurosurgical operations, pathologic anastomosis with nasal cavity, and CSF rhinorrhea, as well as with impaired host defenses . Intravenous ciprofloxacin was given to 20 patients with gram-negative bacillary meningitis . The dose of ciprofloxacin was 200 mg by intravenous infusion, over 30 minutes, every 12 hours for 10 days . Two patients also received intravenous cefotaxime and penicillin G . All patients were monitored daily . Of 20 patients, 18 had cure and therapy failed in two . Because relatively low concentrations of ciprofloxacin are achieved in the CSF, the drug should be used in the treatment of gram-negative bacillary meningitis only if the MICs of the causative pathogen and the ciprofloxacin concentration in CSF can be followed . A randomized, controlled study is needed to compare the efficacy of intravenous ciprofloxacin with standard antibiotic therapy in the treatment of this disease.

Nature, 1989 Nov 16, 342(6247), 296 - 9
Capping and alpha-helix stability; Serrano L et al.; The first and last four residues of alpha-helices differ from the rest by not being able to make the intrehelical hydrogen bonds between the backbone greater than C=O groups of one turn and the greater than NH groups of the next . Physico-chemical arguments and statistical analysis suggest that there is a preference for certain residues at the C and N termini (The C- and N-caps) that can fulfil the hydrogen bonding requirements . We have tested this hypothesis by constructing a series of mutations in the two N-caps of barnase (Bacillus amyloliquefaciens ribonuclease, positions Thr 6 and Thr 26) and determining the change in their stability . The N-cap is found to stabilize the protein by up to approximately 2.5 kcal mol(-1) . The presence of a negative charge of the N-cap adds some 1.6 kcal mol(-1) of stabilization energy because of the interaction with the macroscopic electrostatic dipole of the helix.

J Biol Chem, 1989 Nov 15, 264(32), 19138 - 42
Novel procedures for derivatization of ribosomes for crystallographic studies; Weinstein S et al.; Undecagold and tetrairidium clusters have been used for the preparation of heavy-metal derivatives of ribosomal particles, necessary for the evaluation of phases in the x-ray structure determination of these large particles . To obtain specific binding, monofunctional reagents of the clusters were prepared and were covalently bound to free sulfhydryl groups on the surface of the ribosome . In addition, a mutant of Bacillus stearothermophilus which lacks one ribosomal protein (BL11) was grown . The heavy-atom clusters were covalently bound to isolated protein BL11, and the modified protein was consequently reconstituted into the mutated ribosomal subunits . Crystallographic data have been collected from crystals of native particles, from the mutated ones, and from the iridium- and gold-derivatized subunits . All these crystal forms are isomorphous within the experimental error.

J Biol Chem, 1989 Nov 15, 264(32), 18933 - 8
Amino acid residues stabilizing a Bacillus alpha-amylase against irreversible thermoinactivation; Suzuki Y et al.; The alpha-amylase of Bacillus licheniformis (BLA) is stable and active at high temperature . More than 80% of its activity is retained after heat treatment at 90 degrees C for 30 min, and the optimum temperature for its activity is 80-85 degrees C . In contrast, the alpha-amylase of Bacillus amyloliquefaciens (BAA), the amino acid sequence of which shows 80% homology with that of BLA, is rapidly inactivated at 90 degrees C . Various chimeric genes were constructed from the structural genes for the two enzymes, and their products were analyzed for stability as to irreversible thermoinactivation . Two regions in the amino acid sequence of BLA comprising Gln178 (region I) and the 255th-270th residues (region II), respectively, were shown to determine the thermostability of BLA . Region I plays a major role in determining the thermostability . By means of site-directed mutagenesis of the BAA gene, deletion of Arg176 and Gly177 in region I and substitutions of alanine for Lys269 and aspartic acid for Asn266 in region II were shown to be responsible for the enhancement of the thermostability . Mutant BAAs containing the above deletion and substitutions showed almost the same thermostability as BLA as to irreversible thermoinactivation . Nevertheless, the mutant BAAs showed a temperature optimum as low as that of BAA (65 degrees C), indicating that they are still susceptible to reversible inactivation at temperatures higher than 65 degrees C.

Cancer Res, 1989 Nov 15, 49(22), 6252 - 7
Hyperthermic modulation of respiratory inhibition factor- and iron releasing factor-dependent macrophage murine tumor cytotoxicity; Klostergaard J et al.; Hyperthermia modulated the cytotoxic activities of murine macrophages cocultured with EMT-6 tumor cells, altered the production of the monokines RIF (respiratory inhibition factor) and FeRF (iron-releasing factor) by these effector cells, and perturbed the activities of these monokines against EMT-6 cells . Cytotoxic activities of heated murine macrophages activated by Bacillus Calmette-Guerin were inhibited by heat doses of 40.5 degrees C for greater than or equal to 1 h if heating preceded triggering by the endotoxin lipopolysaccharide; however, cytotoxic activities were better retained if triggering preceded heating by 2 h . Treatment-sequence dependencies were also found in the secretion of some monokines that participate in these macrophage cytotoxic effects . Secretion of both RIF and FeRF by macrophages was slightly augmented or at least better retained if triggering of macrophages occurred several hours before heating for 1 h at 40.5-43 degrees C or for 24 h at 39-40.5 degrees C . If heating was nearly simultaneous with triggering or preceded it by several hours, there was a dose-dependent decrease in monokine secretion . The sensitivity of tumor cell targets to these monokines was also modified by the treatment sequence . When EMT-6 cells were treated with RIF- or FeRF-containing macrophage conditioned supernatants 1 h before heating at 40.5-43 degrees C, the RIF-treated cells became sensitized and the FeRF-treated cells retained better cytotoxic response than if cells had been treated 1 h after heating . Chronic heating for 24 h at 39-40.5 degrees C showed less dependence of tumor cell response on treatment sequence . Similar observations have recently been made in our laboratory for the interaction of tumor necrosis factor cytotoxic pathways with hyperthermia (Klostergaard et al., J . Biol . Response Modif., in press, 1989; Tomasovic et al., Int . J . Hyperthermia, in press, 1989) . Those results and the present observations with the monokines RIF and FeRF support our hypothesis that the cytotoxic actions of macrophages and the cytotoxicity of endogenously added monokines can be augmented by appropriately constructed sequences combining hyperthermia with either macrophage priming/triggering or monokine treatment of tumor cells.

Biochemistry, 1989 Nov 14, 28(23), 9201 - 9
Protection of an unstable reaction intermediate examined with linear free energy relationships in tyrosyl-tRNA synthetase; Wells TN et al.; Linear free energy relationships (LFERs) are powerful tools in the search to understand the relationship between molecular structure and activity . They frequently link the changes in the rate constants for a reaction to changes in the equilibrium constant caused by alterations in structure . In physical-organic chemistry, these have been interpreted to give information on the structure of the transition state . Similar phenomena have been observed for reactions catalyzed by a series of engineered mutants of tyrosyl-tRNA synthetase from Bacillus stearothermophilus . LFERs are applied in this study to probe how the enzyme minimizes its side reactions . A linear free energy relationship is shown between the binding of the unstable enzyme-tyrosyl adenylate complex and its rate constant of hydrolysis . However, mutations of a key residue, His48, show significant deviation from the relationship, implying a role for the side chain in protection of the complex from hydroxide attack . A second linear free energy relationship is shown linking the rate and equilibrium constants for tyrosyl adenylate binding to the enzyme . Four distinct classes of mutation are discussed in the context of this relationship . The data from all but one of these groups of mutations conform well to a linear free energy relationship between the dissociation rate and dissociation equilibrium constants for the enzyme-tyrosyl adenylate complex with slope beta = 1.01 +/- 0.08 . The specificity of binding of tyrosyl adenylate is determined solely by its dissociation rate constant of the intermediate, and the mutations have relatively little effect on the enzyme-tyrosyl adenylate association rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Arch Dermatol, 1989 Nov, 125(11), 1543 - 7
Bacillary (epithelioid) angiomatosis and concurrent Kaposi's sarcoma in acquired immunodeficiency syndrome; Berger TG et al.; Two patients with acquired immunodeficiency syndrome developed simultaneous Kaposi's sarcoma and bacillary (epithelioid) angiomatosis . The distinguishing clinical and histologic features of these two vascular proliferations associated with human immunodeficiency virus disease are described . The lesions of bacillary (epithelioid) angiomatosis contained bacteria, while the lesions of Kaposi's sarcoma did not . With erythromycin therapy, the lesions of bacillary (epithelioid) angiomatosis cleared, while those of Kaposi's persisted . Bacillary (epithelioid) angiomatosis, a treatable but potentially fatal opportunistic infection of human immunodeficiency virus disease, should be considered in the differential diagnosis of vascular lesions in immunosuppressed patients.

Am Rev Respir Dis, 1989 Nov, 140(5), 1189 - 93
Experimental short-course preventive therapy of tuberculosis with rifampin and pyrazinamide; Lecoeur HF et al.; In a first experiment, the efficacy of a 6-month course of isoniazid (INH) alone in comparison with 2-month courses of rifampin (RMP) alone, RMP + pyrazinamide (PZA), or RMP + PZA + INH as preventive therapy of tuberculosis was evaluated in the mouse . To simulate the infected, non-diseased state of humans, a nonreplicating bacillary population of limited size was developed in the mouse . Mice were vaccinated intravenously with 2.74 log10 M . bovis BCG and infected a month later with 3.38 log10 M . tuberculosis . Treatment began 2 wk after infection when the mean size of the M . tuberculosis population was 4.98 +/- 0.26 log10 cfu in the spleen . After 2 months of therapy, the proportion of mice with positive spleen cultures was 100%, 50%, 0%, and 20% in those animals treated, respectively, with INH alone, RMP alone, RMP + PZA, or RMP + PZA + INH . After 6 months of therapy with INH alone, the proportion of mice with positive spleen cultures was 38% . In order to confirm the extreme activity of the combination RMP + PZA and to assess the value of 3 months of therapy with RMP, a second experiment was performed following similar procedures . On completion of treatment, the proportion of mice with positive spleen cultures was 100%, 20%, 0%, and 80% in those animals treated, respectively, for 6 months with INH alone, 3 months with RMP alone, or 2 months with RMP + PZA, or RMP + PZA + INH.(ABSTRACT TRUNCATED AT 250 WORDS)

Urology, 1989 Nov, 34(5), 231 - 7
Conservative treatment of superficial transitional cell carcinoma of prostatic urethra with intravesical BCG; Orihuela E et al.; We reviewed 125 male patients treated with a six-week course of intravesical bacillus Calmette-Guerin (BCG) for superficial transitional cell cancer (TCC) of the bladder that was associated with mucosal involvement of the prostatic urethra in 15 cases . In 13 of these cases, there was a sustained complete response in both the bladder and the prostate . Among the other 110 patients, in 14 TCC developed in the prostate from one to fifty-three (mean 15) months after completion of BCG . Six of these 14 had had complete response in the bladder . Recurrent TCC in the prostate was seen in only 4 percent and 6 percent of patients who had prostatic urethritis on cystoscopy and prostatic granulomas on biopsy, respectively . In contrast, recurrence was observed in 14 percent of those without urethritis and in 44 percent of those without granulomas . Transurethral resection of the prostate (TURP) prior to BCG did not appear to influence tumor recurrence in the prostate . However, after BCG, 3 patients with recurrent TCC in the prostate had complete local response after TURP alone . Further, prostatic urethritis (73%) and prostatic granulomas (60%) were more frequent among patients who had TURP prior to BCG than in those without TURP (33% and 27%, respectively) . Superficial TCC of the bladder associated with mucosal involvement of the prostatic urethra can be treated successfully with intravesical BCG . In addition, our results suggest that intravesical BCG has a prophylactic effect on tumor recurrence in the prostate, and that TURP may have an important role by removing present disease and by facilitating the development of an effective biologic response to BCG in the prostate . TCC in the prostate is a significant cause of relapse, and frequent surveillance of the prostatic urethra should be performed in conservatively treated patients with superficial bladder cancer.

J Urol, 1989 Nov, 142(5), 1376 - 9
Combination immunotherapy of murine transitional cell cancer using BCG and an interferon-inducing pyrimidinone; Sarosdy MF et al.; The immunomodulator ABPP (2-amino-5-iodo-6-phenyl-4(3H) pyrimidinone) is an interferon inducer and has been shown to have in vivo activity against the murine bladder tumor MBT-2 . Two experiments were performed to determine if ABPP might enhance the in vivo anti-tumor activity of Bacillus Calmette-Guerin (BCG) . First, in vivo stimulation of cell-mediated cytotoxicity by BCG and ABPP was measured in C3H mice using a chromium-release assay . An earlier, greater, and longer-lasting increase in cytotoxicity was caused by ABPP than by BCG . Based on the differing times to peak cytotoxic stimulation, groups of 15 mice each were pretreated intraperitoneally at different times prior to inoculation with MBT-2 . Compared to saline, ABPP alone did not increase survival, while BCG alone did increase survival (p less than 0.01), and the combination of BCG and ABPP yielded the highest survival (p less than 0.001) . These results indicate that 1) ABPP affects the immune system differently than BCG, and 2) while ABPP may have less single-agent activity against MBT-2 than BCG, ABPP may serve to potentiate the activity of BCG.

Mikrobiologiia, 1989 Nov-Dec, 58(6), 915 - 9
{Thrombolytic activity of Bacillus mesentericus at different conditions of nitrogen nutrition of a culture}; Cherkesova GV et al.; The work is concerned with studying the effect exerted by different sources of nitrogen nutrition on the biosynthesis of proteinases with a thrombolytic activity by a variant of Bacillus mesentericus, strain 64, obtained with the aid of analytical selection . Protein substrates taken as a nitrogen source stimulate the synthesis of proteinases by the bacterial culture . These enzymes have a high caseinolytic and thrombolytic activity, and the level of their activity correlates with the amount of a protein substrate added to the medium . Ammonium acetate and succinate are the best stimulants for the formation of proteinases when the salts of mineral and organic acids are used as a source of nitrogen nutrition . In that case, the enzymes have a high thrombolytic activity and a low caseinolytic activity . A semi-synthetic medium with the aforementioned nitrogen-containing compounds as a source of nitrogen nutrition is proposed for the synthesis of thrombolytic proteinase by the variant of B . mesentericus.

Bol Med Hosp Infant Mex, 1989 Nov, 46(11), 724 - 7
{Meningitis caused by Bacteroides fragilis in children}; Goyo-Rivas J et al.; Bacteroides fragilis is an obligated anaerobic bacillus which forms part of the normal intestinal flora of the colon and is often seen as a common pathogen in intraabdominal infections . It is an infrequent pathogen in cases of meningitis; a review of the literature reports only eight cases of this disease in children, especially in neonates with conditioning factors such as abdominal sepsis, chronic middle ear otitis and atrial-ventricular derivations . A case of a newborn baby girl with lumbosacral myelomeningocele is reported . After the defect was surgically corrected, the wound became infected, the stitches opened, the child began to have fever, became irritable and suffered convulsions . The spinal tap showed changes compatible to bacterial meningitis, the bacteria was grown on Shaedler medium . The child was treated with cefotaxime and amikacin showing no satisfactory improvement . Afterwards, a second spinal tap showed Bacteroides fragilis.

Trends Biochem Sci, 1989 Nov, 14(11), 450 - 4
Barnase and barstar: two small proteins to fold and fit together; Hartley RW; Barnase and barstar are the extracellular ribonuclease and its intracellular inhibitor produced by Bacillus amyloliquefaciens . Both are small single-chain proteins and thus are suitable for application to the study of how a protein's sequence directs its fold . Barnase has neither disulfide bonds nor non-peptide components and unfolds reversibly in what closely approximates a two-state reaction . The genes for both these proteins have been cloned in E . coli . Expression of barstar is necessary to counter the lethal effect of expressed active barnase . Site-directed mutagenesis is being used to answer specific and general questions relating to protein folding and protein-protein interaction.

Rev Infect Dis, 1989 Nov-Dec, 11 Suppl 7, S1669 - 90
Drug delivery to the central nervous system: general principles and relevance to therapy for infections of the central nervous system; Scheld WM; Drug delivery to the central nervous system (CNS) is of vital concern to the therapy for primary CNS disorders and the development of drug neurotoxicity . The factors influencing drug entry into the CSF include the status of the blood-brain barrier (BBB) and lipid solubility, molecular weight, pKa, protein binding, and removal of the drug from the CSF by an exit pump in the choroid plexus . The most important of these factors is the status of the complex BBB systems . The morphologic equivalent of the BBB and its specialized functions (e.g., transport of D-glucose, amino acids, and ions) are discussed in depth . Methods developed for increasing drug delivery to the CNS by circumvention and/or manipulation of the BBB have included direct injection into the CSF, administration of prodrugs or chemical delivery systems, or reversible "opening" of the BBB by hyperosmotic agents, pentylenetetrazole, etoposide, DMSO, or other agents . The relevance of these general principles to selected examples of CNS infections (i.e., gram-negative aerobic bacillary meningitis and subacute encephalopathy associated with AIDS) is emphasized.

Rev Infect Dis, 1989 Nov-Dec, 11 Suppl 7, S1564 - 71
Monoclonal antibodies for treatment of gram-negative infections; Young LS et al.; Monoclonal antibody technology has permitted researchers to dissect out the protective antibody response to conserved regions of gram-negative bacillary lipopolysaccharides (endotoxins) . Some anticore antibodies can bind to lipid A and have a neutralizing, but not opsonic, activity; these antibodies are usually IgM . IgG antibodies to outer core regions may be weakly opsonic . The outcome of animal protection studies is critically dependent on the choice of challenge organism, dose, timing and rate of antibody administration, and additional factors such as antimicrobial therapy . Protective activity against a wide variety of gram-negative bacillary challenges with the IgM anticore and lipid A reactive antibody, which we have designated E5, is reviewed . Protection in a therapeutic model is demonstrable when the antibody is used in conjunction with appropriate antimicrobial therapy . This antibody is now being assessed in clinical trials . Optimal use of monoclonal antibodies may involve a "cocktail" of antibodies with complementary binding specificities.

J Am Acad Dermatol, 1989 Nov, 21(5 Pt 2), 1119 - 22
Granulomatous eruption after BCG vaccine immunotherapy for malignant melanoma; Shea CR et al.; In a patient receiving intralesional bacille Calmette-Guerin vaccine injections for recurrent cutaneous metastases of malignant melanoma of the right leg, a striking eruption developed on that limb only . The eruption consisted of multiple 1 to 2 mm, reddish brown, flat-topped papules, with confluence into plaques . Histopathologic examination showed multiple tuberculoid granulomas within the papillary and reticular dermis; special stains and cultures were negative for mycobacteria.

Clin Immunol Immunopathol, 1989 Nov, 53(2 Pt 1), 202 - 11
IgA antibodies against phenolic glycolipid I from Mycobacterium leprae in serum of leprosy patients and contacts: subclass distribution and relation to disease activity; Schwerer B et al.; The anti-PGL-I IgA response against phenolic glycolipid I (PGL-I) a specific surface antigen of Mycobacterium leprae, was demonstrated to be essentially of the IgA1 subclass in sera from leprosy patients and contacts . Anti-PGL-I IgA1 mean levels were found to increase significantly from the tuberculoid toward the lepromatous pole of the leprosy disease spectrum, thus resembling the predominating anti-PGL-I IgM response . Furthermore, anti-PGL-I IgA1 values were shown to increase significantly with increasing bacillary load, measured as bacillary index (BI) from skin biopsies . However, a number of BI negative leprosy patients recorded elevated anti-PGL-I IgA1 levels possibly reflecting a persistence of disease activity . Three of 28 household or family contacts of leprosy patients were detected seropositive for anti-PGL-I IgA1 . Thus, our results suggest that anti-PGL-I IgA1 may be considered as an additional parameter for the early detection of infection with M . leprae.

Int J Food Microbiol, 1989 Nov, 9(3), 157 - 65
The microbial ecology of tape ketan fermentation; Ardhana MM et al.; The growth of fungi, yeasts and bacteria was followed during the fermentation of tape ketan . The tape was prepared using samples of Indonesian ragi as inoculum . Fermentation was characterized by the dominant growth of Amylomyces rouxii and Candida pelliculosa (10(5)-10(7) cfu/g), and a lesser contribution from Saccharomyces cerevisiae . Hansenula anomala grew to a limited extent during some fermentations . Bacteria of the genera Bacillus and Acetobacter contributed also to the fermentation, producing populations up to 10(5) cfu/g . Ragi was the main source of microorganisms involved in the fermentation . Tape fermented by inoculating pure cultures of Amylomyces rouxii and the yeasts, either individually or as mixtures, was not of typical quality, indicating the importance of bacteria in the overall fermentation.

J Cell Sci, 1989 Nov, 94 ( Pt 3), 601 - 8
Action of activated 27,000 Mr toxin from Bacillus thuringiensis var . israelensis on Malpighian tubules of the insect, Rhodnius prolixus; Maddrell SH et al.; The action of activated 27,000 Mr toxin from Bacillus thuringiensis var . israelensis (Bti toxin) on Malpighian tubules of Rhodnius prolixus has been investigated . Its binding to the tubules is slowed by low temperature but is not prevented even at 0 degree C . The binding is less effective at pH 10 than at pH7 . Pretreatment of the tubules with 0.1 mmol l-1 ouabain or bumetanide or 1 mumol l-1 5-hydroxytryptamine did not affect the toxicity of the toxin . The toxin causes very large changes in the trans-epithelial potential difference; it changes from 40 mV, lumen negative, often to more than 100 mV, lumen positive . This reflects an initial collapse of the potential of the basal cell membrane, followed by a large positive-going potential change at the luminal cell membrane . Just prior to the effects of the toxin on rapid fluid secretion, the basal cell membrane becomes permeable to sucrose molecules . Raffinose at 170 mmol l-1 in the bathing solution does not protect the tubules from Bti toxin action but dextran, Mr5000, at 60 mmol l-1 significantly delayed failure of fluid secretion and, even more, the onset of staining of the tubule cells with Trypan Blue . Exposing tubules to saline that is calcium-free and/or magnesium-free, or has a composition adjusted to be similar to that of the intracellular milieu, does not affect the time course of failure of fluid secretion induced by the toxin . There is no evidence that effective aggregates of Bti toxin molecules are formed in concentrated solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Bol Med Hosp Infant Mex, 1989 Nov, 46(11), 709 - 14
{Neonatal systemic infection caused by Listeria monocytogenes}; Solorzano-Santos F et al.; Listeria monocytogenes is a gram positive cocco-bacillus which causes perinatal infections and also attacks immunocompromised hosts . Little is known about it in our medium . As part of a prospective study on neonatal systemic infections, its participation at the National Institute of Perinatology was researched . During a period of 18 months, 9,283 live newborns were observed, 141 of them were diagnosed with neonatal septicemia . During this period seven neonates had systemic infections due to Listeria monocytogenes: three had septicemia (two of these with meningitis) and all seven cases had pneumonia . The gestational age of the neonates was 26.1 to 41 weeks (X + DS = 35 + 4.3), with a weight of 830 g to 2,975 g (X + DS 1,958 + 773), four out of seven weighed less than 2,000 grams . The most frequent clinical manifestation was respiratory related causing a need for a differential diagnosis with hyaline membrane disease, transitory tachypnea and meconium swallowing at birth . All of the strains isolated were found to be susceptible to ampicillin, penicillin, gentamicin and amikacin; requiring high CMI levels of cephalosporins.

Fiziol Zh SSSR Im I M Sechenova, 1989 Nov, 75(11), 1554 - 9
{The age-related characteristics of the effect of vasopressin on the vascular smooth muscles of the human brain}; Gurevich MI et al.; Vasopressin was shown to cause a concentration-dependent relaxation of human basilar {correction of bacillary} artery's smooth muscles . In old age, the reactivity of these muscles in decreased and tachyphylaxis to vasopressin disappeared . This seems to be connected with a decrease in the quantity of smooth muscle's V2-inhibitory receptors . Protein kinase C activation does not affect molecular mechanisms of vasopressin action on human basilar {correction of bacillary} artery's smooth muscles . The data obtained suggest vasopressin participation in regulation of human cerebral blood flow and a limitation of adaptive properties of brain vessels' smooth muscles in old age . These factors, apparently, play a role in development of the brain circulation pathology in old men.

Biokhimiia, 1989 Nov, 54(11), 1894 - 903
{Isolation, purification and properties of restrictase and methylase BstN1 from Bacillus stearothermophilus}; Baryshev MM et al.; Restriction-methylation enzymes BstN1 from Bacillus stearothermophilus were isolated and purified . These enzymes are related to a new class of restriction-methylation enzymes of the second type, whose modifying component is N4-cytosine-DNA-methylase . Both enzymes recognize the DNA sequence CC(A/T)GG . Restrictase BstN1 is a protein made up of one subunit with a molecular mass of 25 kDa . The molecular mass of native DNA-methylase BstN1 is about 55 kDa . The temperature optima for restrictase and methylase BstN1 are around 60 degrees C . Possible uses of BstN1 restriction-methylation enzymes for the analysis of cytosine methylation in bacterial and higher plant DNA are discussed.

J Toxicol Sci, 1989 Nov, 14(4), 237 - 45
Subacute toxicity of (-)15-deoxyspergualin in BALB/c mice . I . Hematological study; Okubo M et al.; (-)15-Deoxyspergualin (DSP), a bacterial metabolite obtained from Bacillus laterosporus, was shown to effectively suppress autoantibody production in mice and alloimmune reaction in mice and rats . Its potential toxicity and safety margin in BALB/c mice were studied by administering the drug, 0.5-5.0 mg/kg body weight, s.c., 6 days/week for 3 months . The animals were weighted every two weeks, and their blood was drawn for hematological and liver function studies . Control animals showed a gradual increase in body weight from 17.8 +/- 1.1 g (SD) to 20.2 +/- 3.2 g during the 3 months of treatment . The body weight of 0.5 mg- and 2.5 mg-DSP group was not significantly different from that of the control group . In contrast, 5.0 mg-DSP group of animals showed significant decrease in their weight compared with the control after 5 weeks of treatment (P less than 0.01) . They were then sacrificed . WBC in 0.5 mg-, 2.5 mg- and 5.0 mg-DSP groups were significantly lower compared with the control after 3 months of treatment . On the other hand, only 5.0 mg-DSP group showed significantly lower level in their RBC, Hb and Hct values (P less than 0.05) . In contrast, platelet counts in 2.5 mg- and 5.0 mg-DSP group significantly increased . No manifest effect due to DSP on differential count of leukocytes was observed, although on day 92, a significant difference was shown in the stab cells of the 0.5 mg-DSP group and in the eosinophils in the 2.5 mg-DSP.(ABSTRACT TRUNCATED AT 250 WORDS)

Bioorg Khim, 1989 Nov, 15(11), 1569 - 72
{Derivatives of daunorubicin containing an inosine fragment}; Olsuf'eva EN et al.; Condensation of daunorubicin or its (13 R, S)-dihydro derivative with inosine dialdehyde in the presence of NaBH3CN yielded novel derivatives of anthracycline antibiotics with incorporated inosine residue: 3'-deamino-3'-{(2" R)-(hypoxanthyl-9)-(6" S)-hydroxymethylmorpholino-N4"}- daunorubicin and (13 R,S)-dihydro-3'-deamino-3'-{(2" R)-(hypoxanthyl-9)-(6" S)- hydroxymethylmorpholino-N4"}-daunorubicin . The compounds did not inhibit growth of Bacillus mycoides and were less cytotoxic in vitro and less toxic in vivo than the parent antibiotics.

Appl Environ Microbiol, 1989 Nov, 55(11), 2779 - 88
Binding and aggregation of the 25-kilodalton toxin of Bacillus thuringiensis subsp . israelensis to cell membranes and alteration by monoclonal antibodies and amino acid modifiers; Chow E et al.; The 25-kilodalton toxin of Bacillus thuringiensis subsp . israelensis binds irreversibly to Aedes albopictus cells, Choristoneura fumiferana cells, and erythrocytes . The binding to cells increased with both toxin concentration and time and when the cells were first preincubated with unlabeled toxin . Binding data indicated a two- to threefold increase in the rate of binding after the amount of the membrane-bound toxin reached approximately 3.5 fmol/3 x 10(5) A . albopictus cells or 3.3 . fmol/2 x 10(5) C . fumiferana cells . When this level of bound toxin was reached, the toxins also began forming aggregates at the cell membrane . The toxin aggregates were extracted with 10% Triton X-100 and separated from the monomers with a 5 to 20% sucrose density gradient . The toxin aggregates isolated from A . albopictus and C . fumiferana cell membranes were ca . 400 kilodaltons, while those isolated from human erythrocytes were significantly smaller . The proportion of the toxin found in aggregate form increased rapidly with the amount of toxin bound; however, the molecular size of the aggregates remained constant . Eleven monoclonal antibodies raised against the native form of the toxin blocked 80 to 97% of the toxin binding to cells . The epitope of one of these monoclonal antibodies was mapped to a domain which included the cysteine, suggesting the importance of the domain around this amino acid to binding . Toxin binding and cell lysis were also inhibited by treating the toxin with HgCl2, further indicating the importance of the C-terminal hydrophobic cysteine-containing domain in cytolytic activity of the 25-kilodalton protein.

Mikrobiol Zh, 1989 Nov-Dec, 51(6), 54 - 6
{The collagenolytic activity of bacteria of the genus Bacillus isolated from different ecological sources}; Slabospitskaia AT et al.; The ability to hydrolyze collagen was studied in 262 cultures of spore-forming aerobic bacteria isolated from the soil, human and animal organism, from insects and also of collection strains . 53% of the studied cultures possessed the collagenolytic activity . The most active strains are isolated from the human and animal organism.

Nippon Saikingaku Zasshi, 1989 Nov, 44(6), 789 - 96
{A selective inhibitor of cAMP-dependent protein kinase isolated from an alkalophilic strain of Bacillus species}; Murakami K et al.; Many alkalophilic bacteria were found to produce inhibitors of protein kinases . We isolated a novel inhibitor of protein kinase from an alkalophilic strain of Bacillus species . This substance was A heat-stable peptide with a molecular weight of 13,000 daltons . It was found to be a selective inhibitor of cyclic AMP-dependent protein kinase (A kinase) . The inhibition of a kinase by this substance was non-competitive with histone or ATP . It behaved distinctly; other known inhibitors such as H-7, H-8, Staurosporine, K-252 and Erbstatine inhibit protein kinase less selectively and their functions are competitive with either substrate or ATP . This inhibitor was found to bind to the regulatory subunits of A kinase and markedly inhibited the separation of the catalytic subunits from A kinase induced by the binding of cAMP despite of no effect on the binding of cAMP . Thus, the activation step of A kinase was influenced by this inhibitor . This molecule had no effect on the inhibition by cAMP of CHO cell proliferation . This may have been due to the inability of this molecule to reach the target in the cell . Modification of the molecule itself or the administration method is needed for cellular or animal application.

Mol Microbiol, 1989 Nov, 3(11), 1533 - 43
Specificity of Bacillus thuringiensis for lepidopteran larvae: factors involved in vivo and in the structure of a purified protoxin; Arvidson H et al.; The relative LD50 values in two test Lepidoptera of Bacillus thuringiensis subspecies kurstaki HD1, which contains three crylA protoxin genes, was the same as a plasmid-cured derivative or a Bacillus cereus transcipient containing only one of the three genes . Differential rates of transcription of these genes in the original strain could account, at least partly, for this result . Strains containing only the single protoxin gene (crylA(b} produced inclusions when grown at 25 degrees C but not 32 degrees C, despite transcription of this gene at both temperatures . The instability of the crylA(b) protoxin was not found in the parental B . thuringiensis subsp . kurstaki HD1 strain grown at either temperature, however, so kurstaki HD1 strains with multiple protoxin genes must produce some stabilizing factor, perhaps another protoxin . The cryl protoxins contain a highly conserved carboxyl half which is proteolytically removed upon conversion to toxin . All of the protoxin cysteines are present in protease-sensitive regions and they are oxidized in inclusions . Most of the disulphides appear to be essential for specificity since their reduction in the crylA(b) protoxin resulted in loss of selectivity for one of the test insects . This lack of specificity was also found for this protoxin produced by an Escherichia coli clone, probably because of the reducing conditions in these cells . Specificity was restored by reoxidation of the pure protoxin, by removal of the carboxyl half of oxidized protoxin with trypsin, or by subcloning of the toxin portion . The oxidized form of protoxins must be important for specificity, for the formation of crystalline inclusions, and probably for interactions required for the stabilization of some protoxins.

J Gen Microbiol, 1989 Nov, 135 ( Pt 11), 3035 - 41
A novel phage genome integrated into a plasmid in Bacillus thuringiensis strain AF101; Kanda K et al.; Bacillus thuringiensis strain AF101 possesses a single plasmid (pAF101) with a molecular size of 42 MDa (69 kb) . During plasmid curing experiments in strain AF101, we found that a phage (J7W-1) was induced by ethidium bromide treatment . Moreover, the phage genome (48 kb) hybridized only with pAF101 on a Southern blot of the DNA of a cleared lysate prepared from strain AF101 . Comparison of the restriction patterns of pAF101 and J7W-1 phage DNA revealed that pAF101 contains not only the entire phage DNA but also a plasmid-specific DNA region . These results indicate that the J7W-1 genome has been stably integrated into pAF101 in strain AF101 . Integration of the J7W-1 genome into a plasmid was also observed after phage infection of the type strain of B . thuringiensis subsp . israelensis.

J Gen Microbiol, 1989 Nov, 135 ( Pt 11), 2813 - 8
Sporulation of a bacitracin-sensitive mutant of Bacillus licheniformis is self-inhibited by bacitracin; Podlesek Z et al.; A mutant of Bacillus licheniformis (BLU166) sensitive to its own antibiotic bacitracin was isolated and the mutation bcr-l was mapped close to the bacitracin synthetase genes . The sensitivity was shown to be specific for bacitracin . Two further bacitracin-sensitive strains were constructed, one (BLU171) with normal ability to synthesize bacitracin, and one (BLU170) a bacitracin non-producer . In addition to an increased sensitivity of growing cells to bacitracin, sporulation of the mutant strain BLU171 was self-inhibited by bacitracin . It is concluded that (1) there might exist at least two levels of resistance to bacitracin; (2) mutation bcr-1 affects a 'structural' component, which may protect the sensitive reaction of cell-wall biosynthesis; (3) sporulation is affected to a greater extent by bacitracin than vegetative growth; and (4) synthesis of bacitracin is independent of the presence of this resistance mechanism since the sensitive mutant produces similar amounts of the antibiotic to the wild-type strain.

Rev Infect Dis, 1989 Nov-Dec, 11 Suppl 7, S1629 - 43
Tropical disease in the immunocompromised host: melioidosis and pythiosis; Tanphaichitra D; Melioidosis is an infection of humans and animals caused by a gram-negative motile bacillus, Pseudomonas pseudomallei . Forty-nine patients with melioidosis complicating diabetes mellitus, collagen vascular disorders, leukemia/lymphoma, and other hematologic malignancies are described . Twenty-nine of these patients had disseminated/septicemic infection, two developed toxic shock syndrome, and one with AIDS experienced recrudescent melioidosis . Patients with disseminated melioidosis often have a variety of defects in cellular immunity both in vitro and in vivo . In humans with recrudescent melioidosis, cellular immunity can be transferred by a transfer factor and by levamisole, a cellular immunopotentiating agent . The results of the treatment of our patients with disseminated/septicemic melioidosis with antimicrobial agents in combination have been successful . In recent years, four cases of fungal arteritis due to Pythium species and one case of keratitis due to Pythium were seen . Almost all patients with fungal arteritis had thalassemia; all presented with pain in the lower extremities and gangrenous lesions of the toes . Pythium species, an aquatic Phycomycetes, was identified in these cases as a human pathogen on the basis of clinical features, pathologic findings, and--of greatest importance--the isolation of the etiologic fungi . These five cases with remarkably similar presentations exhibited certain similarities with and differences from cases of mucormycosis, entomophthoromycosis, and peniciliosis.

Biochem J, 1989 Nov 1, 263(3), 665 - 70
Kinetic study of the irreversible thermal denaturation of Bacillus licheniformis alpha-amylase; Violet M et al.; The irreversible thermal inactivation of Bacillus licheniformis alpha-amylase was studied . A two-step behaviour in the irreversible denaturation process was found . Our experimental results are consistent only with the two-step model and rule out the two-isoenzyme one . They suggest that the deactivation mechanism involves the existence of a temperature-dependent intermediate form . Therefore the enzyme could exist in a great number of active conformational states . We have shown that Ca2+ is necessary for the structural integrity of alpha-amylase . Indeed, dialysis against chelating agents leads to a reversible enzyme inactivation, though molecular sieving has no effect . Further, the key role of Ca2+ in the alpha-amylase thermostability is reported . The stabilizing effect of Ca2+ is reflected by the decrease of the denaturation constants of both the native and the intermediate forms . Below 75 degrees C, in the presence of 5 mM-CaCl2, alpha-amylase is completely thermostable . Neither other metal ions nor substrate have a positive effect on enzyme thermostability . The effect of temperature on the native enzyme and on one intermediate form was studied . Both forms exhibit the same optimum temperature . Identical activation parameters for the hydrolytic reaction catalysed by these two forms were found.

Agents Actions, 1989 Nov, 28(3-4), 264 - 71
Cathepsin B and D activity in alveolar macrophages from rats with pulmonary granulomatous inflammation or acute lung injury; Lesser M et al.; Cathepsin B and D activity was determined using specific synthetic substrates in alveolar macrophages (AMs) obtained from Sprague-Dawley rats with experimentally induced pulmonary granulomatous inflammation . Increased cathepsin B activity was found 4 days after intravenous injection of complete Freund's adjuvant (CFA), but not after injection of live bacillus Calmette-Guerin organisms (BCG), indicating that the enzyme response was unrelated to the subsequent development of granulomatous inflammation . Findings of comparable increases in enzyme activity following injection of mineral oil (MO) indicate that the response to CFA was due to the oil component . Significantly, oleic acid (OA), a natural fatty acid, did not stimulate enzyme activity although the agent, like MO, caused acute lung injury as assessed by 125I albumin uptake . At 7 and 28 days following injection of CFA, cathepsin B levels in AMs were the same as those in animals given normal saline (NS), although bronchoalveolar lavage (BAL) samples still contained increased numbers of AMs, and cells obtained at 28 days phagocytosed more polystyrene microspheres . Cathepsin D activity did not increase 4 days after injection of CFA or BCG + CFA; at 28 days following injection of BCG + CFA activity was significantly decreased as compared to animals given NS . The data reveal a differential response of two lysosomal enzymes during the early phases of granulomatous inflammation.

Pediatr Infect Dis J, 1989 Nov, 8(11), 763 - 7
Detection of circulating antibodies to purified protein derivative by enzyme-linked immunosorbent assay: its potential for the rapid diagnosis of tuberculosis; Barrera L et al.; An enzyme-linked immunosorbent assay for detecting antibodies to purified protein derivative was evaluated as a rapid method for serodiagnosis of childhood tuberculosis . Its specificity for IgG antibodies was 0.98 as determined in 55 sera from nontuberculous children who showed no significant effect of previous Bacillus Calmette-Guerin vaccination on the production of specific antibodies . Results were negative in 29 of 33 (87.9%) tuberculin-positive children and in 18 of 20 (90.0%) contacts, none of whom had evidence of tuberculosis . The sensitivity of this test was 0.51 as determined in 49 sera from bacteriologically confirmed cases; 17 of 27 smear positive cases and 8 of 22 children with positive cultures were detected . Results were positive in 32 of 114 (28.1%) patients with a diagnosis of tuberculosis not confirmed by microbiology . Consequently whereas a negative result does not rule out tuberculosis, a positive result is a strong indication of the disease . The IgM antibody determination yielded much less discriminative results.

J Hosp Infect, 1989 Nov, 14(4), 339 - 50
The effect of air on the moist-heat resistance of Bacillus stearothermophilus spores; Scruton MW; The presence of air in an autoclave chamber and load is generally considered to reduce the lethal effect of the process on bacterial spores . In this study, the heat inactivation of spores of Bacillus stearothermophilus (NCIB 8224) was measured in the presence and absence of air at 100% relative humidity . The results confirm that air significantly enhances the lethality of moist-heat on this strain of B . stearothermophilus.

Antibiot Khimioter, 1989 Nov, 34(11), 841 - 6
{The role of bitoxibacillin components in the manifestation of its biological activity}; Efimtsev EI et al.; New methods were developed and applied to quantitative determination of beta-exotoxin and antibiotic activity of delta-endotoxin with respect to Micrococcus spp . in bitoxibacillin (BTB) and the fermentation broths prepared under industrial conditions . The biosynthesis of beta-exotoxin in the period of its maximum accumulation during the fermentation was estimated . It was shown that the primary biological effect of BTB on insects consisted in the actions of beta-exotoxin and delta-endotoxin . Biological activity of each of the entomocidal components of the entomocidal components of BTB did not practically correlated with the number of viable spores . There was a correlation between the antibiotic activity of crystalline B . thuringiensis subsp . thuringiensis solutions and the insecticidal activity of the entomopathogenic preparations . Determination of beta-exotoxin and antibiotic activity of delta-endotoxin might be used as a complex procedure for testing the quality of BTB . The method for estimating antibiotic activity of the crystal solutions allowed one to assay the biological activity of other preparations based on Bacillus thuringiensis non-synthesizing beta-exotoxin.

APMIS, 1989 Nov, 97(11), 981 - 6
Collaboration between human blood dendritic cells and monocytes in antigen presentation; Rasanen L et al.; We compared human blood dendritic cells and monocytes for their capacity to produce secreted and membrane interleukin 1 (IL-1), stimulate mixed leukocyte reaction (MLR) and augment microbial antigen-induced T lymphocyte proliferation . Our enriched dendritic cell and monocyte fractions contained greater than 80% and greater than 93% dendritic cells and monocytes, respectively . Monocytes produced about ten times higher amounts of membrane and secreted IL-1 than dendritic cells, which in turn were more potent in presenting HLA-DR antigens in MLR . Both accessory cell types presented purified protein derivative of tuberculin (PPD) equally well, whereas monocytes were better with fixed Bacillus Calmette Guerin (BCG) bacteria . Processing of BCG was chloroquine-sensitive . Coculture experiments suggested that there was collaboration or synergy between dendritic cells and monocytes in antigen processing and presentation.

Mikrobiol Zh, 1989 Nov-Dec, 51(6), 30 - 4
{The effect of the cultivation conditions on lectin production by representative bacteria in the genus Bacillus}; Podgorskii VS et al.; Dynamics of the lectin activity accumulation in the culture liquid was studied through the example of certain representatives of spore-forming aerobic bacteria in the process of their growth . The environmental factors (the temperature, pH value, the presence of definite carbohydrates in the medium) were investigated for their effect on the biosynthesis of extracellular lectins by producers . Based on these studies it was possible to establish optimal conditions of cultivation and to obtain the high specific lectin activity of the culture liquid for each strain.

Biochem J, 1989 Nov 1, 263(3), 905 - 12
Trapping the acyl-enzyme intermediate in beta-lactamase I catalysis; Cartwright SJ et al.; Cryoenzymology techniques were used to facilitate trapping an acyl-enzyme intermediate in beta-lactamase I catalysis . The enzyme (from Bacillus cereus) was investigated in aqueous methanol cryosolvents over the 25 to -75 degrees C range, and was stable and functional in 70% (v/v) methanol at and below 0 degree C . The value of kcat . decreased linearly with increasing methanol concentration, suggesting that water is a reactant in the rate-determining step . In view of this, the lack of incorporation of methanol into the product means that the water molecule involved in the deacylation is shielded from bulk solvent in the enzyme-substrate complex . From the lack of adverse effects of methanol on the catalytic and structural properties of the enzyme we conclude that 70% methanol is a satisfactory cryosolvent system for beta-lactamase I . The acyl-enzyme intermediate from the reaction with 6-beta-(furylacryloyl)amidopenicillanic acid was accumulated in steady-state experiments at -40 degrees C and the reaction was quenched by lowering the pH to 2 . H.p.l.c . experiments showed covalent attachment of the penicillin to the enzyme . Digestion by pepsin and trypsin yielded a single labelled peptide fragment; analysis of this peptide was consistent with Ser-70 as the site of attachment.

Biochem J, 1989 Nov 1, 263(3), 883 - 8
Dynamic light-scattering studies on the effect of heat and disinfectants on spores of Bacillus cereus; Molina-Garcia AD et al.; The relative stability of spores of Bacillus cereus grown at three different temperatures was examined by using quasi-elastic light scattering (q.l.s.) in conjunction with turbidity and scanning electron microscopy (s.e.m.) . Cultures grown at 20, 30 and 40 degrees C (BC20, BC30 and BC40 respectively) were compared in terms of (i) their effective hydrodynamic radius, rH, as determined from q.l.s . and (ii) their gross morphology, as determined from s.e.m . The effects of autoclaving at 121.1 degrees C on both these properties was also examined . We observed (1) that cultures BC20 and BC30 appeared to have similar values for rH, whereas that of BC40 appeared some 50% higher, and (2) BC40 had a correspondingly much lower heat resistance (its structural integrity was lost after about 20 min autoclaving, whereas that of BC20 and BC30 was retained even after 80 min autoclaving) . These data were in good agreement with independent measurements of heat-resistance coefficients . Changes in the hydrodynamic radius, polydispersity (both using q.l.s.) and turbidity were monitored with time on addition of the disinfectants sodium hypochlorite and peracetic acid; again BC40 appeared to have a lower resistance.

J Bacteriol, 1989 Nov, 171(11), 6077 - 83
Phosphatidylinositol-specific phospholipase C of Bacillus cereus: cloning, sequencing, and relationship to other phospholipases; Kuppe A et al.; The phosphatidylinositol (PI)-specific phospholipase C (PLC) of Bacillus cereus was cloned into Escherichia coli by using monoclonal antibody probes raised against the purified protein . The enzyme is specific for hydrolysis of the membrane lipid PI and PI-glycan-containing membrane anchors, which are important structural components of one class of membrane proteins . The protein expressed in E . coli comigrated with B . cereus PI-PLC in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as detected by immunoblotting, and conferred PI-PLC activity on the host . This enzyme activity was inhibited by PI-PLC-specific monoclonal antibodies . The nucleotide sequence of the PI-PLC gene suggests that this secreted bacterial protein is synthesized as a larger precursor with a 31-amino-acid N-terminal extension to the mature enzyme of 298 amino acids . From analysis of coding and flanking sequences of the gene, we conclude that the PI-PLC gene does not reside next to the gene cluster of the other two secreted phospholipases C on the bacterial chromosome . The deduced amino acid sequence of the B . cereus PI-PLC contains a stretch of significant similarity to the glycosylphosphatidylinositol-specific PLC of Trypanosoma brucei . The conserved peptide is proposed to play a role in the function of these enzymes.

Gene, 1989 Oct 30, 82(2), 237 - 48
Nucleotide sequences of genes encoding heat-stable and heat-labile glyceraldehyde-3-phosphate dehydrogenases; amino acid sequence and protein thermostability; Tesfay HS et al.; The structural gene (gapST) encoding glyceraldehyde-3-phosphate dehydrogenase (GPDH; EC 1.2.1.12) from Bacillus stearothermophilus has been cloned in Escherichia coli using plasmid pBR322 as a vector; the homologous gene (gapCO) from Bacillus coagulans was cloned from a phage lambda library . Expression of the cloned gap genes revealed that, as in the wild-type (wt) organisms, the GPDH from B . stearothermophilus (GPDH-ST) was intrinsically heat stable (hs) and that from B . coagulans (GPDH-CO) heat labile (hl) . The cloned gap genes were sequenced and the deduced amino acid (aa) sequences were found to be highly conserved (91.6% homology), despite the large difference in thermostability between these two enzymes . Of the 28 aa which differ between the two proteins, most of which occur in the middle third of the monomeric subunit, 5 aa involve replacement of alanine in the hl GPDH-CO, by proline in the hs GPDH-ST, and are especially interesting in terms of their potential contributions to thermostability . Conservation at the DNA level is equally dramatic, with the two gap genes exhibiting 93.3% nucleotide sequence homology . These highly expressed genes exhibit an equivalent codon bias, which more closely resembles that of highly expressed E . coli genes, than that of B . stearothermophilus genes whether highly or weakly expressed.

Carbohydr Res, 1989 Oct 23, 192, 83 - 90
Formation of a cycloinulo-oligosaccharide from inulin by an extracellular enzyme of Bacillus circulans OKUMZ 31B; Kawamura M et al.; A strain of Bacillus circulans OKUMZ 31B, isolated from soil, has been shown to produce an extracellular enzyme that converts inulin into cycloinulo-oligosaccharides . The main product was identified as cycloinulo-hexaose . The enzyme is arbitrarily designated as cycloinulo-oligosaccharide fructanotransferase.

Eur J Biochem, 1989 Oct 20, 185(1), 151 - 5
Improved purification and biochemical properties of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis; Kupke T et al.; Monophosphatidylinositol inositol phosphohydrolase (phosphatidylinositol-specific phospholipase C . PtdIns-PLC . EC 3.1.4.10) has been purified from a Bacillus thuringiensis culture supernatant and from the cellular fraction of a recombinant Escherichia coli clone containing the PtdIns-PLC gene from B . thuringiensis . The two-step purification procedure involved ion-exchange chromatography on DEAE-Sepharose followed by separation on a Mono-Q/FPLC-column with yields of 32% and 50%, respectively . The molecular mass was determined to be 34 kDa by SDS/PAGE . The isoelectric point of the enzyme was 5.15 . The amino-terminal sequences were shown to be identical for the enzymes purified from both organisms . PtdIns-PLC was inhibited by divalent cations using mixed micelles of Triton X-100 and pure phosphatidylinositol . PtdIns-PLC activity was detectable on polyacrylamide gels by activity staining on phosphatidylinostiol-containing agarose.

J Mol Biol, 1989 Oct 20, 209(4), 793 - 800
Three-dimensional structure of cyclodextrin glycosyltransferase from Bacillus circulans at 3.4 A resolution; Hofmann BE et al.; Cyclodextrin glycosyltransferase (EC 2.4.1.19) from Bacillus circulans has been purified, crystallized and analyzed by X-ray diffraction . The enzyme is monomeric . SDS/polyacrylamide gel electrophoresis gave an Mr of 73,600(+/- 1000), corresponding to 670(+/- 10) amino acid residues . The structure of the crystalline enzyme has been elucidated at a resolution of 3.4 A, using multiple isomorphous replacement and solvent flattening for phase determination . The resulting electron density map allowed tracing of the polypeptide chain; 664 residue positions have been assigned . The chain fold has been subdivided into five domains . The N-terminal domain forms a (beta alpha)8-barrel, which contains the second domain of about 55 residues as an insert after the third beta-strand . The three remaining domains form almost exclusively beta-pleated sheet structures and consist of about 90, 80 and 95 residues . The chain fold of the three N-terminal domains of 492 residues resembles closely the two known structures of alpha-amylases . This geometric similarity corresponds to the observed amino acid sequence homology . On the basis of the sequence homology with alpha-amylases, the active center can be located . The fourth domain has an immunoglobulin fold and is far away from the active center, while the fifth domain participates in the formation of the broad depression at the active center . Accordingly, the cyclodextrin glycosyltransferase chain fold can be considered as an alpha-amylase chain fold with two additional domains.

Eur J Biochem, 1989 Oct 20, 185(1), 157 - 62
Scalar, dipolar-correlated and J-resolved 2D-NMR spectroscopy of the specific phenolic mycoside of Mycobacterium tuberculosis; Daffe M et al.; Two-dimensional chemical shift correlated (COSY), nuclear Overhauser (NOESY) and J-resolved spectroscopy were used to determine the complete structure of the major triglycosyl dimycocerosyl phenol phthiocerol of the tubercle bacillus (strain Canetti) without any other analytical technique . The COSY spectrum of the native glycolipid allowed the composition of the trisaccharide and the location of one methoxyl group to be determined through the assignment of the resonances of the non-anomeric methine protons . Information with respect to the configuration of the sugar residues, the pyranose structure and the linkage sites of the trisaccharide were obtained by the analysis of the COSY spectrum of the peracetylated glycolipid . The NOESY spectrum confirmed the linkage sites through the inter-residue connectivity across glycosidic linkages and allowed the determination of the sequence of the oligosaccharide and the configuration of its sugar residues from the through-space connectivities . The J-resolved spectroscopy was used to elucidate the structure of the glycolipid and gave additional data showing the presence of 11 non-equivalent methyl groups and allowing the recognition of two other analogues of the phenol phthiocerol derivative in the mixture . Thus, the non-destructive 2D-NMR spectroscopy was found to be satisfactory for the analysis of mycobacterial mycosides.

Biochim Biophys Acta, 1989 Oct 16, 985(2), 233 - 5
Immunoelectron microscopic localization of the F1F0 ATPase (ATP synthase) on the cytoplasmic membrane of alkalophilic Bacillus firmus RAB; Rohde M et al.; Evidence that the F1F0 ATPase (ATP synthase) of alkalophilic Bacillus firmus RAB is localized exclusively on the cytoplasmic membrane was obtained by immunogold electron microscopy using a highly specific polyclonal antibody against the beta subunit of Escherichia coli F1F0 ATPase . The energetic problem faced by cells of B . firmus RAB growing oxidatively at pH 10.5 despite a low protonmotive force across the cytoplasmic membrane cannot, therefore, be circumvented by localization of energy transducing functions on hypothetical internal membranes.

J Biol Chem, 1989 Oct 15, 264(29), 17551 - 8
Two forms of plasma membrane-intercalated heparan sulfate proteoglycan in rat ovarian granulosa cells . Labeling of proteoglycans with a photoactivatable hydrophobic probe and effect of the membrane anchor-specific phospholipase C; Yanagishita M et al.; The structures of cell-associated heparan sulfate (HS) proteoglycans and their interaction with the plasma membrane was studied using rat ovarian granulosa cell culture . HS proteoglycans were either metabolically labeled by incubating cell cultures with {3H} leucine and {35S}sulfate or labeled in plasma membrane preparations with a photoactivatable reagent, 3-(trifluoromethyl)-3-(m-{125I}iodophenyl)diazirine (TID), a compound which has been shown to selectively label the hydrophobic membrane-binding domains of several proteins . After purification of HS proteoglycans from the labeled cell cultures or from the labeled membrane preparations by repeated Q-Sepharose ion exchange chromatography in 8 M urea, they were analyzed by Superose 6 gel filtration and octyl-Sepharose chromatography both in 4 M guanidine HCl . The results indicated that the HS proteoglycans were labeled with 125I and therefore have an intramembranous domain . Phospholipase C (Bacillus thuringiensis), which specifically cleaves phosphatidylinositol membrane anchors, released approximately 25% of the 35S-labeled HS proteoglycans from the cell surface as well as 20-30% of the 125I-label from the 125I-TID-labeled HS proteoglycans . These data indicate that a subpopulation of HS proteoglycans are intercalated into the plasma membrane through a linkage structure involving phosphatidylinositol . Phospholipase C-resistant, 125I-labeled HS proteoglycans represent those species inserted into membrane through an intercalated peptide sequence . Core protein size of phosphatidylinositol-anchored species estimated by polyacrylamide gel electrophoresis after heparitinase digestion was approximately 80 kDa, and it was significantly larger than that of the directly intercalated species (approximately 70 kDa).

Arch Dermatol, 1989 Oct, 125(10), 1380 - 2
Dysgonic fermenter type 2 septicemia with purpura fulminans . Dermatologic features of a zoonosis acquired from household pets; Herbst JS et al.; Dysgonic fermenter type 2, a gram-negative bacillus that is part of the normal oral flora of dogs and cats, is responsible for increasing numbers of cases of fulminant septicemia in humans . Patients usually have preexisting medical illnesses, but infection also occurs in otherwise healthy individuals . Most infections are acquired through animal contact . Dermatologic eruptions occur in half of the patients with dysgonic fermenter type 2 infection, and include petechiae, purpura, cellulitis, and gangrene.

J Urol, 1989 Oct, 142(4), 1073 - 4; discussion 1074-5
Systemic bacillus-Calmette Guerin infection after transurethral administration for superficial bladder carcinoma; Sakamoto GD et al.; It is well known that intravesical bacillus Calmette-Guerin therapy is a safe and effective treatment for superficial bladder carcinoma with few serious complications . However, we report a case of pneumonitis, hepatitis, azotemia, fever and pancytopenia in a patient after transurethral administration of bacillus Calmette-Guerin . Even though severe systemic complications are rare, patients who have high fevers of signs of other systemic manifestations require documentation of the extent of the infection . Also, we address the use of prophylactic antituberculous drugs, not only to prevent these symptoms but also to raise the questions of diminishing the immunological effect of the treatment.

J Bacteriol, 1989 Oct, 171(10), 5296 - 303
Structure, surface charge, and self-assembly of the S-layer lattice from Bacillus coagulans E38-66; Pum D et al.; In freeze-etched preparations, whole cells from Bacillus coagulans E38-66 exhibited an oblique S-layer lattice (a = 9.4 nm; b = 7.4 nm; gamma = 80 degrees) . The three-dimensional structure of the crystalline array was characterized by optical and computer image analysis . The lattice showed two distinctly shaped types of pores . In vitro self-assembly of isolated subunits yielded flat sheets and open-ended cylinders composed of two back-to-back monolayers . Unlike whole cells, in vitro self-assembly products were capable of binding polycationized ferritin (pI, approximately 11) . This showed that only the inner S-layer face adhering to the peptidoglycan-containing layer in whole cells was net negatively charged . S-layer monomers and/or oligomers were capable of generating a closed monolayer with oblique symmetry on poly-L-lysine-coated supports . The monolayer had a typical crazy paving appearance, with numerous crystal boundaries . The handedness of the oblique lattice and ability to bind polycationized ferritin revealed that the subunits had bound with the outer, not net negatively charged face to the poly-L-lysine-coated supports . Carbodiimide-activated carboxyl groups on either cell wall fragments or self-assembly products could covalently bind high-molecular-weight nucleophiles such as ferritin . This confirmed the location of negatively charged carboxyl groups on the outermost surface of both S-layer faces . The difference in pH optimum for carbodiimide activation indicated a preponderance of alpha- and beta-carboxyl groups on the inner S-layer face and a preponderance of beta- and gamma-carboxyl groups on the outer S-layer face.

Med Trop (Mars), 1989 Oct-Dec, 49(4), 337 - 41
{Dapsone inhibition of the bactericidal action of rifampicin on Mycobacterium leprae in mice}; Millan J et al.; In the experimental infection of mice by Mycobacterium leprae, the bactericidal effect of 4 weekly doses of rifampicin (RMP) is completely suppressed if this administration is preceded by a daily treatment of dapsone (DDS) during one month then continued in conjunction with rifampicin . The application of this methodology: the delayed adding of rifampicin clearly shows the bacillary persistence induced by dapsone (DDS) . The rifampicin appears to be less effective on Mycobacterium leprae when its metabolism is inhibited either by the action of a drug such as dapsone (DDS), or spontaneously . The highlighting of this late-appearing antagonism between rifampicin and dapsone in mice, should not at present lead to the questioning of the therapeutic procedures recommended by the WHO, because of the limits of this experimental model, namely the small size of bacillary populations studied over relatively short periods of time.

Indian J Lepr, 1989 Oct, 61(4), 495 - 502
Detection of subclinical infection in leprosy: an 8 years follow-up study; Bharadwaj VP et al.; A follow-up study has been carried out using Fluorescent Leprosy Antibody Absorption (FLA-ABS) test in 1069 healthy contacts of multi and pauci-bacillary leprosy patients . Simultaneously lepromin testing with Dharmendra antigen has also been done to determine their delayed type hypersensitivity . In nearly 8 years of follow-up, 46 contacts have developed disease and of these 41 contacts were FLA-ABS positive and lepromin negative . It is inferred that test (along with lepromin) can be used to identify the contacts who are at higher risk of developing the disease . FLA-ABS test has also been found to be highly sensitive for detection of subclinical infection specially in younger age groups . This test could therefore serve as a very sensitive epidemiological tool for assessing the extent of disease in the community and for monitoring the transmission of disease especially after MDT and other intervention measures.

Biochem J, 1989 Oct 1, 263(1), 65 - 71
Release of penicillinase by Escherichia coli HB101 (pEAP31) accompanying the simultaneous release of outer-membrane components by Kil peptide; Aono R; The plasmid pEAP31 contains an alkaliphilic-Bacillus penicillinase gene and a colicin E1 kil gene . Escherichia coli HB101 carrying pEAP31 grown at high temperature released outer-membrane proteins, lipopolysaccharide and phosphatidylethanolamine into the culture medium . Concurrently, penicillinase that had accumulated in the periplasm of the organism was released from the cells . Phospholipase A1-A2 in the outer membrane was not activated in the organism . The results suggest that the release of accumulated periplasmic penicillinase from the producer cells was caused by partial disruption of the outer membrane mediated by the Kil peptide.

Immunology, 1989 Oct, 68(2), 253 - 9
Adoptive immunity to Mycobacterium bovis strain bacillus Calmette-Guérin by long-term cultured T-cell line in nude mice; Yamamoto S et al.; Tuberculin-active peptide-reactive T-cell lines were established from the popliteal lymph node of BALB/cA mice immunized with heat-killed Mycobacterium tuberculosis to investigate the cellular mechanisms in the protective immunity of tuberculosis . These T-cell lines, consisting mainly of L3T4 surface antigen-positive cells, were transferred intravenously into nude mice infected with M . bovis strain bacillus Calmette-Guerin (BCG) . Four or 6 weeks after transfer, footpad swelling and hepatic granuloma formation were observed and viable counts in the liver were decreased . Reduction of viable counts in the liver was obviously preceded by the hepatic granuloma formation . An effect of Lyt-2+ T cells which might be included in the inoculum could be ruled out by the experiment using T-cell lines pretreated with anti-Lyt-2 monoclonal antibody and complement . These results indicated that T cells required for protective immunity in these experiments belong to the L3T4-positive TDTH subset . However, their protective activity was inferior to that of freshly prepared immune lymph node T cells . From the observation of migration patterns of 51Cr-labelled T cells in BCG-infected nude mice, a relatively high proportion of freshly isolated T cells, but only a small part of T-cell line, deposited in the spleen of infected nude mice . This difference in migration pattern of freshly isolated and cultured T cells could be one of the reasons for the less in vivo anti-bacterial activity of the latter.

Am Rev Respir Dis, 1989 Oct, 140(4), 1007 - 11
Prospective trial of timing of bacillus Calmette-Guérin vaccination in Canadian Cree infants; Pabst HF et al.; We studied 184 Cree Indian infants in randomized, prospective fashion to assess the effect of age on lymphocyte sensitization to purified protein derivative (PPD) before and after and without bacillus Calmette-Guerin (BCG) vaccination . Lymphocyte responses to PPD, Candida, and streptokinase were measured at birth and at intervals later . The mean response of paired values from 26 infants without BCG vaccination rose for the PPD stimulation index (SI) from 2.7 at birth to 3.9 before 2 yr of age . The SI for both Candida and streptokinase for this group of infants rose significantly in the first 2 yr (p less than 0.05) . In 66 infants who received BCG in the first 7 days of life, the PPD-SI rose from 3.1 to 35.3 (p less than 0.001) . In 17 infants who received the vaccine later but before 9 months, it rose from 3.1 at birth to 24.9, and in 14 who received it between 9 months and 2 yr, it rose from 2.2 to 52.9 . The lymphocyte responses to PPD after BCG in these two groups were significantly different (p less than 0.05) . There was no evidence in the older infants that a raised PPD-SI before BCG vaccination affected lymphocyte sensitization by the vaccine . We conclude that increasing the age at vaccination with BCG from birth to more than 9 months enhances immunologic sensitization to PPD significantly in this population.

Antibiot Khimioter, 1989 Oct, 34(10), 761 - 6
{Pharmacokinetic properties of pancreatic and microbial ribonucleases}; Kurinenko BM et al.; Pharmacokinetic properties of pancreatic RNAase (RNAase I), RNAase of Bacillus intermedius (RNAase Bi) and RNAase of Streptomyces rimosus (RNAase Sr) were studied on albino rats . RNAase Bi was shown to be characterized by a higher rate and level of absorption into the systemic blood flow, higher retention time, lower elimination from the kidneys and tissues of the peripheral chamber (skeletal muscles) and higher distribution in the other animal organs such as the heart, spleen and brain . It was concluded by the experimental results that the higher antiviral efficacy of RNAase Bi (RNAase Bi greater than RNAase Sr greater than RNAase I), as was known from the literature data, and the ability to stimulate the immunity correlated with higher biological availability of the enzyme in the animals and could be due to its pharmacokinetic properties.

Rev Clin Esp, 1989 Oct, 185(5), 240 - 5
{Evolution of pulmonary tuberculosis at a specialized hospital from its beginning to the present time . General aspects}; Palomero Gil A et al.; A retrospective analysis of pleuropulmonary tuberculosis patients admitted to a specialized hospital from the day it started functioning to 1986 is performed . The epidemiological and clinical aspects of 966 medical histories are reviewed . 32% of these patients had household contacts with patients suffering the same disease . We could also observe an increase with time in the percentage of diabetic and alcoholic patients . With respect to the bacteriological study we have to point out that in 86% of patients Koch Bacillus was detected . A decrease in length of hospital stay as well as a decrease in mortality up to 3% is observed . These variables are recorded at two time points: when the hospital first started and at the present time, analyzing any possible statistically significant differences.

Appl Environ Microbiol, 1989 Oct, 55(10), 2517 - 21
Transformation of Bacillus polymyxa with plasmid DNA; Mallonee DH et al.; A plasmid transformation system was developed for Bacillus polymyxa ATCC 12321 and derivatives of this strain . The method utilizes a penicillin-treated-cell technique to facilitate uptake of the plasmid DNA . Low-frequency transformation (10(-6) per recipient cell) of plasmids pC194, pBD64, and pBC16 was accomplished with this method . Selection for the transformants was accomplished on both hypertonic and nonhypertonic selective media, with the highest rates of recovery occurring on a peptone-glucose-yeast extract medium containing 0.25 M sucrose . Several additional plasmids were shown to be capable of transferring their antibiotic resistance phenotypes to B . polymyxa through the use of a protoplast transformation procedure which allowed for a more efficient transfer of the plasmid DNA . However, cell walls could not be regenerated on the transformed protoplasts, and the transformants could not be subcultured from the original selective media.

Nippon Hinyokika Gakkai Zasshi, 1989 Oct, 80(10), 1459 - 65
{Comparative study on prophylactic intravesical instillation of bacillus Calmette-Guerin (BCG) and adriamycin for superficial bladder cancers}; Tachibana M et al.; In this study we evaluated 77 patients with superficial bladder cancer who were treated with either intravesical bacillus Calmette-Guerin (BCG) (44 patients) or doxorubicin hydrochloride (Adriamycin, ADR) (33 patients) for prophylaxis of tumor recurrence after transurethral resection . The estimated actuarial probability of non-recurrence rate at three years for the BCG group was 73.0%, and the actuarial non-recurrence rate for the ADR group was 27.3%; a statistically significant difference (p = 0.0013) . A comparison between the two groups was also made according to the patient's background, including whether the tumor was initial or recurrent, solitary or multiple, and the tumor grade . In all areas of the study, except for grade-1 tumors, the BCG group was significantly superior to the ADR group . The efficacy of BCG therapy as a result of different BCG treatment protocol was evaluated for six weekly instillations 1) without further maintenance instillation, 2) followed by 12 months of maintenance, and 3) followed by more than 18 months of maintenance . Long-term maintenance BCG instillation group (more than 18 months) showed the most favorable results, however, the differences were not statistically significant . These results indicate that intravesical BCG instillation was significantly superior to ADR in the prevention of bladder cancer recurrence and that six weekly intravesical BCG instillations may provide adequate prophylactic effects against recurrence of superficial bladder cancers.

FEMS Microbiol Lett, 1989 Oct 1, 52(1-2), 165 - 9
Transformation of Bacillus amyloliquefaciens by electroporation; Vehmaanpera J; A method for transformation of whole Bacillus amyloliquefaciens cells by electroporation was developed . The procedure is as efficient as the protoplast transformation method, resulting in up to 10(5) transformants/micrograms plasmid DNA, but requires less effort and time . Cells for electroporation were grown to late exponential phase in a rich medium supplemented with 0.25 M sucrose, washed with and resuspended in 0.25 M sucrose, 1 mM HEPES, 1 mM MgCl2, 10% (v/v) glycerol, pH 7.0, at 3-5 x 10(10) cells/ml for storage at -80 degrees C . The highest transformation frequency was obtained at 7.5 kV/cm with a 25 microF capacitor . The transformation efficiency increased linearly with DNA concentration at least over the range 10 ng-12.5 micrograms/ml . Transformations with ligated DNA and of industrial strains were also successful . In addition, B . subtilis cells treated as above could be transformed by electroporation, resulting in 10(4) transformants/micrograms DNA at 12.5 kV/cm.

In Vitro Cell Dev Biol, 1989 Oct, 25(10), 887 - 91
Resistance to the Bacillus sphaericus toxin in cultured mosquito cells; Schroeder JM et al.; Increasing doses of Bacillus sphaericus toxin were used to select a toxin-resistant cell line from the Culex quinquefasciatus line . This resistant cell line was proven to be C . quinquefasciatus in origin by isozyme analysis and karyotype . The resistant line bound fluorescent-labeled toxin as did the unselected susceptible line . A high level of resistance was quickly achieved, and this level was maintained after 4 mo . culture in the absence of toxin.

Antibiot Khimioter, 1989 Oct, 34(10), 739 - 41
{An immunoenzyme method of isolation of foot-and-mouth disease virus by using beta-lactamase conjugate with virus-specific antibodies}; Mishchenko VA et al.; It was shown that in was feasible to use conjugates of virus-specific antibodies and beta-lactamase from Bacillus licheniformis 749/c to identify aphthosa virus antigens . The antigen titers determined by enzyme immunoassay (EIA) using a beta-lactamase conjugate were 5-64 times higher than the analogous indices of the complement fixation test . Unlike EIA, that by using the antibody conjugates with peroxidase or alkaline phosphatase there were observed no "background" responses.

Hum Pathol, 1989 Oct, 20(10), 1025 - 6
Infection-associated vascular lesions in acquired immunodeficiency syndrome patients; Abrams J et al.; Several reports have recently appeared in the literature describing "unique" non-neoplastic vascular lesions in patients with the acquired immunodeficiency syndrome (AIDS) . These lesions may be mistaken clinically and histologically for Kaposi's sarcoma . The terms epithelioid angiomatosis, epithelioid or histiocytoid hemangioma, and pyogenic granuloma have all been used to describe a similar entity in which cat scratch disease bacillus (CSDB) was subsequently identified . Lesions closely resembling this entity occur in patients with bartonellosis . We report a case of a cutaneous vascular lesion on the hand of an AIDS patient in which cytomegalovirus (CMV) and organisms consistent with CSDB were both found . Simultaneous infections with CMV and CSDB have not been previously described . The presence of these organisms in and around endothelial cells may provide the common stimulus for the formation of these reactive vascular proliferations.

Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 1989 Oct, 11(5), 358 - 62
{Multiplication of mycobacterium leprae in nude mice}; Wang HY; This article reported the multiplication of mycobacterium leprae in immune-deficient animals-nude mice (NIH/nu/nu) . Mycobacterium leprae taken from multibacillary leprosy patient were inoculated with 1.0 x 10(4)/foot pad into right hind foot pad(FP) of nude mice and CFW mice . The results of bacterial enumeration showed that acid-fast bacillus (AFB) in the nude mice continuously multiplied . At 21 months after inoculation, the numbers of AFB increased by over 108,000-folds and reached greater than 1.08 x 10(9)/FP (greater than 5.4 x 10(10)/g tissues), while the CFW mice developed only 3.68 x 10(6)/FP and exhibited their characteristic limited growth pattern . It is indicated that the phase of logarithmic growth was prolonged in nude mice . The AFB multiplied were identified as mycobacterium leprae by following identification test: (1) failure to growth on artificial media; (2) loss of acid-fastness by pyridine extraction; (3) typical growth curve of mycobacterium leprae in foot-pad and sensitivity to antileprosy drugs . The histopathological examination of nude mice foot-pad inoculated were found lepromatous lesions, in which enormous numbers of AFB were seen on 476 days postinoculation, AFB disseminated into the fear, nose, sciatic nerve, testicle, lymphnodes, liver, spleen, lung, and noninoculated foot-pad . As mentioned above, it is considered that experimental leprosy was established in nude mice.

J Gen Microbiol, 1989 Oct, 135 ( Pt 10), 2735 - 41
Characterization of soluble fibronectin binding to Bacille Calmette-Guérin; Aslanzadeh J et al.; Fibronectin (FN), a 420 kDa glycoprotein, consists of two similar subunits linked by a disulphide bond near the C-terminal end . FN is present in soluble and matrix forms in various body fluids and tissues and has been shown to bind to variety of organisms . We characterized the conditions required for 125I-FN binding to Bacille Calmette-Guerin (BCG) . The binding was dose-dependent, reached saturation within 3 min, and was essentially irreversible for at least 24 h under optimal binding conditions at pH 6.0 . In contrast, the binding was reversible (greater than 90% in 24 h) when the pH was increased to 10.0 . Scatchard analysis of the dose-response experiments produced a straight line, suggesting the presence of a single class of FN receptor on BCG . 125I-FN binding was trypsin-sensitive, suggesting that the BCG-binding molecule is a protein . The number of FN receptors was determined to be 8000-15,000 per bacterium . 125I-FN binding was pH dependent, with maximal binding at acidic pH . 125I-FN binding was sensitive to the presence of NaCl, with 0.08 M-NaCl inhibiting binding by 85% . These data demonstrate that soluble FN binds to a trypsin-sensitive cell-surface component of BCG in an essentially irreversible manner.

J Gen Microbiol, 1989 Oct, 135 ( Pt 10), 2717 - 22
Surface hydrophobicity of spores of Bacillus spp; Koshikawa T et al.; The surface hydrophobicity of 12 strains of Bacillus spp . was examined in a hexadecane-aqueous partition system . Mature and germinated spores of Bacillus megaterium QM B1551 transferred to the hexadecane layer, while vegetative and sporulating cells did not . Wild-type spores were more hydrophobic than spores of an exosporium-deficient mutant of B . megaterium QM B1551, although the mutant spores were shown to be hydrophobic to some extent by using increased volumes of hexadecane . This result suggests that the exosporium is more hydrophobic than the spore coat and that the surface hydrophobicity of spores depends mainly on components of the exosporium . The surface hydrophobicity of spores of nine other species of Bacillus was also examined, and spores having an exosporium were more hydrophobic than those lacking an exosporium . Thus measurement of the hydrophobicity of spores by the hexadecane partition method may provide a simple and rapid preliminary means of determining the presence or absence of an exosporium.

Biol Chem Hoppe Seyler, 1989 Oct, 370(10), 1139 - 43
Proteolytic specificity of the neutral zinc proteinase from Bacillus mesentericus strain 76 determined by digestion of an alpha-globin chain; Stoeva S et al.; The proteolytic specificity of the neutral zinc proteinase from Bacillus mesentericus strain 76 (MCP 76)/Bacillus subtilis was determined by using the alpha-chain of walrus hemoglobin as substrate . The resulting peptides were fractionated by gel filtration and than isolated by reversed-phase HPLC . The peptides were identified on the basis of their amino-acid compositions and aligned with the known sequence of the walrus alpha-chain . The proteolytic specificity of MCP 76, deduced from the experimental cleavage pattern is compared to that of thermolysin . The amino-acid residues in positions P1 and P'1 on both sides of the scissible bond are considered as most important for the cleavage . MCP 76 prefers leucine, valine, phenylalanine and threonine in position P'1 as well as lysine, threonine, leucine and alanine in position P1 and thus differs from thermolysin which shows no preference for threonine in P'1 and accepts numerous amino-acid residues of different type in P1.

Appl Environ Microbiol, 1989 Oct, 55(10), 2595 - 600
Toxin production by Bacillus cereus dairy isolates in milk at low temperatures; Christiansson A et al.; A total of 136 strains of Bacillus cereus isolated from milk and cream were evaluated for toxin production based on HeLa S3, Vero, and human embryonic lung (HEL) cell cytotoxicity in vitro . HEL cell monolayers were more susceptible than the other two cell lines . The percentage of isolates exhibiting HEL cytotoxicity was similar (43.0 and 48.4%) when the strains were grown in brain heart infusion broth containing 0.1% glucose (BHIG) at 7 and 24 h, respectively, at 30 degrees C . In milk, only 21.8% of isolates showed HEL cytotoxicity at 7 h, and the number increased significantly to 73.2% at 24 h at 30 degrees C . Further, 102 toxin-positive isolates were acclimatized to grow at 8 degrees C in milk . Ninety-four (92.2%) of the strains produced HEL cytotoxicity of various degrees with no strict correlation to bacterial cell numbers and also elicited vascular permeability reaction in rabbit skin . Under aerated growth conditions (agitation, 200 rpm) B . cereus elicited cytotoxicity in BHIG and in milk at temperatures of 30, 15, and 8 degrees C . However, in nonaerated (stagnant) cultures toxin production was diminished (BHIG) or completely lost (milk) at all temperatures . Toxin production at 8 degrees C was evaluated in two different types of commercial cardboard milk packages by inoculation with a potent toxigenic dairy isolate . No detectable HEL cytotoxicity was observed in milk in any of the packages either at stagnant conditions or during mechanical shaking . However, the same strain produced cytotoxin in whipped cream at 8 degrees C.

J Bacteriol, 1989 Oct, 171(10), 5702 - 6
Apparent phosphate retrieval system in Bacillus cereus; Guddal PH et al.; Bacillus cereus secretes three different phospholipases C . We studied the effect of Pi levels in the growth medium on the production of these exoenzymes . Production of both phosphatidylcholine-preferring phospholipase C and sphingomyelinase C was repressed by Pi in the growth medium, whereas production of phosphatidylinositol phospholipase C was unaffected . We also found that B . cereus secretes a phosphate-repressed alkaline phosphatase activity . Together with a previously reported highly efficient, active uptake system for Pi, these three phosphate-repressed exoenzyme activities seem to be part of a phosphate retrieval mechanism that operates under growth-limiting concentrations of Pi . In natural soil systems, which are the natural habitats of B . cereus, the scarcity of Pi is the major growth-limiting factor . A phosphate-repressed metalloprotease activity was also detected in culture supernatants of B . cereus . It is unclear whether this exoenzyme activity also participates in the proposed phosphate-scavenging system.

Proc Natl Sci Counc Repub China B, 1989 Oct, 13(4), 258 - 61
Bacereutin, an antifungal antibiotic isolated from metabolites of Bacillus cereus CHU 130; Tschen JS et al.; An antifungal metabolite, bacereutin, was isolated from culture filtrate of Bacillus cereus CHU 130 . The bacterium was isolated from soils collected in Changhwa County, Taiwan, and was grown in soybean meal-mannitol broth for production of the antibiotic metabolite . The antibiotic metabolite was isolated by adsorption column chromatography of Amberite XAD-2 and was purified by passing through the chromatographic columns packed with Dowex 50W-X8, Sephadex LH 20 and Biogel P-2 . The antibiotic metabolite was soluble in water and 87% acetone, and was slightly soluble in methanol, but was not dissolved in n-propanol, n-butanol, acetone, benzene and ethyl acetate . The antibiotic metabolite was a heat-stable and ninhydrin-positive substance . The antibiotic activities of bacereutin were tested by means of the agar-diffusion plate method . The antibiotic metabolite inhibited the growth of Saccharomyces cerevisiae CHU 1, Paecilomyces variotii CHU 6, Rhizomucor miehei CHU 40 and Fusarium oxysportum CHU 98 . Bacereutin was a ninhydrin-positive antifungal antibiotic.

J Urol, 1989 Oct, 142(4), 978 - 81; discussion 981-2
Expression of blood group precursor T antigen as a prognostic marker for human bladder cancer treated by bacillus Calmette-Guerin and interleukin-2; Dow JA et al.; The relationship between T antigen expression in transitional cell carcinoma of the bladder and response to bacillus Calmette-Guerin plus interleukin-2 treatment was studied . A total of 25 patients received combined treatment with bacillus Calmette-Guerin and interleukin-2 at weekly instillations for 6 consecutive weeks and then monthly for 1 year . T antigen expression in all patients was studied before treatment . Of the patients 16 had positive T antigen expression in the tumors: 13 (81%) remained free of tumor for an average of 28 months, 3 had a noninvasive recurrence each at 8 to 27 months (mean 18 months) and all responded well to a repeated treatment cycle . Nine patients were negative for T antigen: 1 (11%) remained free of tumor for 11 months, while 8 had recurrence within 12 months . Of the latter 8 patients 2 had noninvasive recurrence each at 4 to 5 months but they responded to a repeated treatment cycle and remained free of tumor for 4 to 22 months (mean 13 months), 4 had subsequent repeated recurrences at 12 to 41 months (mean 22 months) and 2 had progression to deep invasion resulting in cystectomy at 11 to 12 months . Thus, the disease-free rate (continuous complete response) in patients with tumors positive for T antigen was 81%, while that in patients with tumors negative for T antigen was 11% (p less than 0.005, chi-square test) . The over-all response rate in patients with positive and negative tumors to bacillus Calmette-Guerin and interleukin-2 treatment was 100 and 33%, respectively, and that for the total patients was 76% . T antigen may serve as a target recognition structure for effector cells generated by bacillus Calmette-Guerin plus interleukin-2 treatment . This study suggests that T antigen is useful to predict the response of bladder tumors to treatment with bacillus Calmette-Guerin and interleukin-2.

FEBS Lett, 1989 Sep 25, 255(2), 361 - 4
Purification of a DNA methyltransferase from Bacillus natto B3364; Kim EL et al.; An S-adenosyl-L-methionine:DNA-methyltransferase, termed M.BnaI, was purified from Bacillus natto B3364 strain by successive column chromatography . The molecular weight determined by gel filtration was 37 kDa for M.BnaI . Analysis of methyltransferase by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed correspondence of the M.BnaI activity with one protein band at a molecular weight of 35 kDa . Sequencing of pUC19 DNA methylated with M.BnaI showed the cytosine-5 methylation in the BnaI recognition sequence GGAT decreases CC at the position indicated by the arrow.

FEBS Lett, 1989 Sep 25, 255(2), 237 - 40
Investigation of neutral endopeptidases (EC 3.4.24.11) and of neutral proteinases (EC 3.4.24.4) using a new sensitive two-stage enzymatic reaction; Indig FE et al.; A sensitive two-stage enzymatic reaction for mammalian and bacterial metalloendopeptidases has been developed using the substrate 3-carboxypropanoyl-alanyl-alanyl-leucine-4-nitroanilide supplemented with Streptomyces griseus amino-peptidase . Neutral endopeptidase EC 3.4.24.11 from bovine kidney hydrolyzes the substrate (pH 7.5, 25 degrees C) with a catalytic efficiency (kcat = 1.2 x 10(2) s-1, Km = 0.15 mM) of the highest ever reported for the enzyme acting on synthetic chromophoric and fluorogenic substrates . Thermolysin hydrolyzes the substrate at a faster rate (kcat = 1.2 x 10(3) s-1) but the overall efficiency is diminished by a higher Km (4.2 mM) . Suspensions of human neutrophil cells and culture filtrates of Bacillus cereus have been assayed sensitively for their neutral endopeptidases and neutral proteinase activities, respectively . The assay provides a convenient tool for the kinetic investigation of neutral endopeptidases and neutral proteinases and for assessing their function in biological systems.

Anal Biochem, 1989 Sep, 181(2), 212 - 5
Isolation of carboxyl-terminal peptides from proteins by diagonal electrophoresis: application to the entomocidal toxin from Bacillus thuringiensis; Bietlot HP et al.; A procedure for the selective isolation of the C-terminal peptides from enzymatic digests of proteins is described . The methodology is based on the diagonal electrophoretic procedure described by R . G . Duggleby and H . Kaplan (1975) Anal . Biochem . 65, 346-354) . The carboxyl groups in the protein are amidated with {14C}-methylamine followed by enzymatic digestion . Since only the C-terminal peptides lack a free carboxyl group, these peptides will lie on a diagonal line of a two-dimensional electrophoretogram run at pH 2.1 and 4.4 . The diagonal line is delineated by autoradiography using {14C}taurine (net charge = 0 at pH 2.1 and 4.4) and {14C}choline (net charge = +1 at pH 2.1 and 4.4) . Radioactive C-terminal peptides lie between these markers and can be directly excised for analysis . This procedure permits the detection and selective isolation of C-terminal peptides with minimal losses . The procedure was applied to the test proteins alpha-chymotrypsin and ribonuclease A . It was used to determine the C-terminus of the Bacillus thuringiensis toxin generated by tryptic cleavage of the protoxin.

Ther Drug Monit, 1989 Sep, 11(5), 574 - 9
Serial pharmacokinetic dosing of aminoglycosides: a community hospital experience; Hoffa DE; The aminoglycoside antibiotics are useful in the treatment of gram-negative bacillary infections but are potentially toxic . A method to maximize their therapeutic benefit while minimizing their risk of toxicity is desirable . Serial pharmacokinetic dosing has been proposed as a method to achieve these goals . An audit was conducted comparing optimal dosing of aminoglycosides by physicians of a community hospital using a serial pharmacokinetic dosing service versus its nonuse . Optimal dosing was 81% (81/101) of trough-peak pairs using the service versus 17% (28/161) not using the service (p less than 0.001) . This difference was due to greater achievement of therapeutic peak levels with the service (96%, 97/101) than without (32%, 52/161) . There was no difference in toxic trough level occurrence, with 15% (15/101) occurrence with the service and 17% (27/161) occurrence without its use (p greater than 0.05) . Clinical nephrotoxicity occurred 0% (0/49) of the time with the use of the service versus 7% (6/88) of the time without its use (p greater than 0.05) . The data presented here demonstrate that serial pharmacokinetic dosing of aminoglycosides results in the achievement of therapeutic peak levels in most patients . An understanding of aminoglycoside nephrotoxicity and pharmacokinetics allows the subsequent adjustment of dosing, if necessary, to avoid nephrotoxicity in most patients.

Pediatr Infect Dis J, 1989 Sep, 8(9), 625 - 9
Mycobacterium tuberculosis infection in foreign born adoptees; Lange WR et al.; Korean adoptees placed by Associated Catholic Charities of Baltimore between January, 1985, and December, 1988 (N = 873), underwent surveillance for tuberculosis (TB) . Overall tuberculin reactivity or active TB was documented in nine placements . Two children had active disease; both were skin test-negative and one died of tuberculous meningitis . The median age of the seven asymptomatic tuberculin reactors, none of whom had received Bacillus Calmette-Guerin vaccination, was 1 year . This reactivity rate is approximately 50 times greater than that of age-matched United States-born children . We conclude that high risk pediatric groups for TB, in addition to refugees, displaced persons and black and Hispanic minorities, include children involved in international adoption . Total reliance cannot be placed on screening tuberculin skin tests, and TB must be included in the differential diagnosis of high risk children presenting with recurrent pneumonia, fever of unknown origin, failure to thrive and altered mental status.

Cesk Epidemiol Mikrobiol Imunol, 1989 Sep, 38(5), 311 - 7
{The concept of information systems}; Tichacek B et al.; On the example of existing information systems of communicable diseases the authors analyze the possibilities of and draw attention to the obstacles of their wider use . The following information systems are involved: ARO (acute respiratory diseases), APO (acute diarrhoeal diseases), ISBT (information system of bacillary tuberculosis), ISPO (information system of communicable diseases), ISID (information system of child immunization) . In the general evaluation the authors draw attention to the demand to do away with the scattered character of the system and the necessity of delimitation of the responsibility for the establishment and operation of the information systems . The authors indicate the future desirable development.

J Urol, 1989 Sep, 142(3), 719 - 22
Long-term results of intravesical bacillus Calmette-Guerin therapy for superficial bladder cancer; Sarosdy MF et al.; Between January 1978 and February 1984, 120 patients with superficial bladder tumors and/or carcinoma in situ were enrolled in previously reported therapeutic trials of bacillus Calmette-Guerin . Of all treated patients 78 per cent responded to initial therapy, with a followup of 13 to 120 months (median 67 months) . Of the 18 patients who failed 10 were treated with repeat, intensified courses . Nine patients who had recurrent tumors within 3 to 30 months after initiation of bacillus Calmette-Guerin therapy (median 6 months) eventually ceased having recurrence . Status free of disease in the 9 patients ranged from 25 to 90 months since the last recurrence (median 64 months) . With retreatment of some of the early failures, the initial success rate of 78 per cent was increased to 89 per cent . These data support the concept that intravesical immunotherapy with bacillus Calmette-Guerin should be repeated in patients who initially appear not to respond . The data also suggest that bacillus Calmette-Guerin induces a durable beneficial response rather than simply delays eventual tumor recurrence.

J Bacteriol, 1989 Sep, 171(9), 4881 - 7
Gene cluster containing the genes for tyrocidine synthetases 1 and 2 from Bacillus brevis: evidence for an operon; Mittenhuber G et al.; From a genomic library of the tyrocidine producer Bacillus brevis ATCC 8185 constructed in the bacteriophage vector EMBL3, a recombinant phage which contains the structural genes coding for tyrocidine synthetases 1 and 2, TycA and TycB, was identified . The location of the tycA gene within the 16-kilobase insert of this clone, EMBL25-1, was mapped by hybridization studies by using the previously isolated tycA DNA as a probe . Restriction analyses, the construction of subclones, and the analysis of proteins encoded by the subclones located the tycB gene at the 3' end of the tycA gene and revealed that the two genes are transcribed in the same direction . Nuclease S1 protection studies and DNA sequencing studies of the intergenic region indicated that tycA and tycB are separated by a 94-base-pair noncoding region and suggested that these genes are organized as an operon.

Rev Cubana Med Trop, 1989 Sep-Dec, 41(3), 333 - 40
{Monitoring of the treatment effectiveness in patients with leprosy using solid-phase immunoenzyme assay}; Gonzalez-Abreu E et al.; Serum samples from 184 lepers are studied by means of solid phase ELISA with the semisynthetic antigen disaccharide-bovine albumin analogous of phenolic glycolipid I of Mycobacterium leprae . Patients are grouped according to the clinical presentation of the disease and course of time elapsed since the institution of therapy . Bacilloscopic examinations were made also in 116 patients who were positive at diagnosis . For serologic testing, absorbance values over 0,160 were considered positive . Results in multibacillary patients showed a gradual and significant decrease both of mean absorbance values and in the seropositive ratio in connection with the length of treatment . It was also found that phenolic anti glycolipid antibodies increased with the value of the bacteriologic index . This system is useful in monitoring the effectiveness of chemotherapy in multibacillary leprosy.

Vrach Delo, 1989 Sep, (9), 112 - 4
{The role of swimming pools in the spread of intestinal infections}; Bondarenko VI et al.; A kindergarten swimming bath was experimentally infested with M-17 Bacillus coli . It was found that after bathing 10% of children showed Escherichia M-17 in the feces . It is concluded that presence of conditionally pathogenic microorganisms in the water of swimming baths may be considered a qualitative and quantitative criterion of epidemic risk during bathing in infected basins in case hygienic norms are not observed per person.

Arch Inst Cardiol Mex, 1989 Sep-Oct, 59(5), 511 - 6
{Tuberculous myocarditis . Presentation of a case and review of the literature}; de Leon-Pena A et al.; We report a case of a 22 year-old drug addict man with overt heart failure and cardiomegaly . The patient had no prior history of tuberculosis and not clinical features to suggest this diagnosis . Tuberculosis myocarditis was found at the necropsy . It is important to point out the frequency of this pathology, the uncommon clinical presentation of this case; as well as the lack of immunologic response to the tuberculous bacillus.

J Surg Oncol, 1989 Sep, 42(1), 33 - 8
Advances in the diagnosis and management of bladder cancer: flow cytometry, intravesical bacillus Calmette-Guerin, and systemic chemotherapy; Badalament RA et al.; Recent advances in the diagnosis and treatment of patients with bladder cancer include flow cytometry, intravesical bacillus Calmette-Guerin (BCG), and systemic chemotherapy . Once the subjects of clinical research protocols, these modalities are currently being incorporated into the general management of patients with bladder cancer . Flow cytometry, intravesical BCG, and systemic chemotherapy will be individually reviewed and their role in the management of patients with bladder cancer will be discussed.

Int J Hyperthermia, 1989 Sep-Oct, 5(5), 625 - 39
Temporal dependence of hyperthermic augmentation of macrophage-TNF production and tumor cell-TNF sensitization; Tomasovic SP et al.; Hyperthermia in the therapeutic (greater than or equal to 42-43 degrees C) and febrile (less than or equal to 39-40.5 degrees C) ranges modulated the cytotoxic activities of macrophages cocultured with tumour cells and of the monokine tumour necrosis factor (TNF) against tumour cells . These modulatory interactions had clear treatment-sequence dependencies, and some sequences markedly augmented cytotoxic activities . Heated murine bacillus Calmette-Guerin-activated macrophages retained their cytotoxic activities better in coculture with unheated tumour cells if triggering with the endotoxin lipopolysaccharide preceded 1 h of heating at 42 or 43 degrees C than they did if heating to the same extent was concomitant with, or preceded, triggering . Retention of cytotoxicity in coculture during 24 h of 39 or 40.5 degrees C heating was less dependent on pre-heating triggering . The triggering/heating sequence also had modulatory effects on the secretion by heated macrophages of TNF which is involved in cytotoxic manifestations in coculture . Production of TNF by macrophages heated for 1 h at 40.5-43 degrees C or 24 h at 39 or 40.5 degrees C was augmented 1.5- to 6-fold (depending on the heat dose) when triggering preceded heating, whereas sequences in which heating was concomitant with triggering or preceded triggering were detrimental to TNF secretion . Profound treatment-sequence dependencies were also seen when timing of the addition of recombinant human TNF was varied in relation to the heat treatment of tumour cells . Sensitization of TNF-responsive L-929 and TNF-resistant EMT-6 tumour cells occurred if monokine addition preceded heating, whereas the reverse treatment-sequence reduced or eliminated sensitization . Both tumour cell types were also sensitized to TNF if monokine treatment preceded 24 h heating at 40.5 degrees C . These results support the hypothesis that appropriately constructed sequences for either macrophage priming/triggering or monokine treatment of tumour cells, combined with hyperthermia, could augment the cytotoxic actions of macrophages and the cytotoxicity of endogenously added monokines.

Infect Dis Clin North Am, 1989 Sep, 3(3), 613 - 24
Imipenem and aztreonam; Sobel JD; To date, clinical use of aztreonam has been remarkably restrained and widespread enthusiasm for addition to hospital formularies similarly moderate . Its most accepted indication appears to be in gram-negative bacillary infections in the penicillin-allergic individual where one might ordinarily use an extended spectrum penicillin or a third generation cephalosporin . In all individuals its clinical efficacy appears similar to that of the aminoglycosides, not clinically superior but offering a higher therapeutic index by virtue of its lack of nephrotoxicity and ototoxicity . Its second application, therefore, is as an aminoglycoside-sparing agent even for multiresistant gram-negative bacillary infections, and although by no means inexpensive it offers an efficacious and safe alternative to aminoglycosides in the elderly and other patients with renal insufficiency . Aztreonam can be used effectively in combination with non-beta-lactam agents for mixed infections, even though it is not synergistic in vitro with vancomycin and clindamycin.

J Bacteriol, 1989 Sep, 171(9), 5232 - 6
Molecular cloning of the structural gene for alkaline elastase YaB, a new subtilisin produced by an alkalophilic Bacillus strain; Kaneko R et al.; Alkaline elastase YaB is an extracellular serine protease of the alkalophilic Bacillus strain YaB . We cloned the structural gene, ale, and determined the nucleotide sequence . The mature enzyme (268 amino acids) was preceded by a putative signal sequence and a prosequence (27 and 83 amino acids, respectively) . The mature enzyme was 55% homologous to subtilisin BPN' . Almost all the positively charged residues are predicted to be on the surface of the molecule, which would facilitate binding to elastin . The P1 substrate site-related sequences differed between alkaline elastase YaB and subtilisin BPN'.

J Bacteriol, 1989 Sep, 171(9), 4871 - 5
Cloning, nucleotide sequence, and expression in Escherichia coli of the Bacillus stearothermophilus peroxidase gene (perA); Loprasert S et al.; The gene encoding a thermostable peroxidase was cloned from the chromosomal DNA of Bacillus stearothermophilus IAM11001 in Escherichia coli . The nucleotide sequence of the 3.1-kilobase EcoRI fragment containing the peroxidase gene (perA) and its flanking region was determined . A 2,193-base-pair open reading frame encoding a peroxidase of 731 amino acid residues (Mr, 82,963) was observed . A Shine-Dalgarno sequence was found 9 base pairs upstream from the translational starting site . The deduced amino acid sequence coincides with those of the amino terminus and four peptides derived from the purified peroxidase of B . stearothermophilus IAM11001 . E . coli harboring a recombinant plasmid containing perA produced a large amount of thermostable peroxidase which comigrated on polyacrylamide gel electrophoresis with the B . stearothermophilus peroxidase . The peroxidase of B . stearothermophilus showed 48% homology in the amino acid sequence to the catalase-peroxidase of E . coli.

Southeast Asian J Trop Med Public Health, 1989 Sep, 20(3), 421 - 7
Long residual activity of Bacillus sphaericus 1593 against Culex quinquefasciatus larvae in artificial pools; Pantuwatana S et al.; The microbial larvicide, Bacillus sphaericus 1593, was evaluated against Culex quinquefasciatus larvae in artificial pools at Huay Kwang area in Bangkok between March and October 1985 . This study was aimed at determining the residual activity of B . sphaericus 1593 in waste water under field conditions . The larvicidal activity of B . sphaericus 1593 was found to persist for at least 5 months in artificial pools containing waste water . The populations of B . sphaericus 1593 in the test water fluctuated, decreasing by approximately 4.25 and 3.47 log10 colony forming unit (cfu)/ml from the original concentrations in 60 and 80 days after application and then increasing approximately 2.92 and 2.77 log10 cfu/ml in 92 and 72 days for pools No . 1 and No . 2, respectively . This evidence indicates that B . sphaericus 1593 can recycle in such conditions.

Indian J Med Res, 1989 Sep, 89, 314 - 21
Growth & toxicity of Bacillus thuringiensis var israelensis; Desai SY et al.; Chemically defined media containing glutamic acid, glutamine and aspartic acid at a 20 mM concentration individually supported abundant growth and sporulation of Bacillus thuringiensis var israelensis . The parasporal crystals produced in these media were toxic to fourth instar larvae of Culex pipiens quinquefasciatus Say . The medium containing 20 mM of glutamine induced early sporulation of B . thuringiensis var israelensis . An LC50 value of 3.92 micrograms/l was obtained . Calcium alginate and agar agar immobilized formulations of Bacillus thuringiensis var israelensis exhibited toxicity of 90 per cent and above to C . pipiens quinquefasciatus Say larvae, over a period of 90 days.

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