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 bl