Bacillus

Int J Pept Protein Res, 1991 Apr, 37(4), 325 - 30
Modification of a zinc proteinase from Bacillus mesentericus strain 76 by diethylpyrocarbonate; Stoeva S; Diethylpyrocarbonate (DEPC) inactivated the neutral zinc proteinase from Bacillus mesentericus strain 76/Bacillus subtilis (MCP 76) by ethoxycarbonylation completely . Exposure of the enzyme to DEPC together with the competitive inhibitor Z-L-phenylalanine prevented the loss of activity toward both peptide and protein substrates . Treatment with hydroxylamine restored the catalytic properties of the modified MCP 76 to that of the native enzyme . After chymotryptic digestion of ethoxycarbonylated MCP 76 in the presence and absence of Z-L-phenylalanine a single histidyl residue essential for the enzyme activity was isolated and identified as histidine 231.

Ann Plast Surg, 1991 Apr, 26(4), 403 - 6
Spontaneous regression of metastatic melanoma; Hurwitz PJ; Spontaneous regression of malignant melanoma is a well-known but rare event . It seems to be caused by immunological enhancement similar to the regression observed after intralesional injection of Bacille Calmette-Guerin into cutaneous melanoma metastases . A patient is presented in whom an incisional biopsy of a metastatic melanoma in an inguinal lymph node was followed by complete disappearance of the metastasis . Reviewing the literature, several similar patients were found in whom spontaneous regression of a metastatic melanoma occurred after an incomplete excision or biopsy of the tumor . It appears that in rare instances, an unspecific mechanical stimulus may trigger an increase in immunocompetence, although laboratory evidence for this has only rarely been produced . A plea is made for serial immunological studies in consecutive patients with melanoma, to further elucidate these puzzling phenomena.

J Vet Med Sci, 1991 Apr, 53(2), 281 - 6
Purification and characterization of the vascular permeability factor produced by Bacillus cereus; Shinagawa K et al.; Purification of an extracellular protein exhibiting the vascular permeability activity produced by Bacillus cereus was performed by ammonium sulfate precipitation followed by chromatography on DE-32 cellulose, Sephadex G-100, and Sephadex G-75 . The purified protein was found to be electrophoretically and antigenically almost homogeneous although it contained a trace of contaminant . The molecular weight of the protein was calculated to be 45,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate . The purified protein showed vascular permeability activity and mouse lethal toxicity, and caused fluid accumulation in ligated mouse intestinal loops, whereas it did not show any hemolytic and lecithinase activities . From these findings, the purified protein is suggested to be an enterotoxin (or a diarrheagenic toxin) responsible for diarrhea caused by B . cereus in a diarrheal-type food poisoning.

J Vet Med Sci, 1991 Apr, 53(2), 167 - 71
Fluid accumulation in mouse ligated intestine inoculated with the vascular permeability factor produced by Bacillus cereus; Shinagawa K et al.; Partially purified vascular permeability (VP) factor (VPF) of Bacillus cereus induced fluid accumulation in the ligated intestinal loops of mouse (MIL) and rabbit (RIL), suggesting that the VP activity may correlate with fluid accumulation in ligated intestinal loops of these animals . Fluid accumulation was observed at 6-8 hr in 55-67% of mouse intestinal loops inoculated with 40-50 immunodiffusion units (IDU) of partially purified VPF, whereas it was found at 2 hr in all loops with 400-600 IDU of partially purified VPF . In rabbit intestinal loops with 120-190 IDU of partially purified VPF, fluid accumulation was observed at 6 hr . From these findings, VPF produced by B . cereus can be easily detected in both MIL and RIL . The intestinal tissue of mouse intestinal loops was histopathologically damaged at different concentrations of the VPF to induce fluid accumulation . With 50 IDU of partially purified VPF, severe edema was found in the laminia proprial layer and submucosa . With 600 IDU of partially purified VPF, on the other hand, severe necrosis in the surface epithelium of villus and laminia proprial layer, and hyperemia in the submucosa were observed, suggesting that partially purified VPF may be cytotoxic and/or intestinecrotic.

Protein Expr Purif, 1991 Apr-Jun, 2(2-3), 199 - 204
Purification and partial characterization of acyl carrier protein from Euglena gracilis variety bacillaris; Williams SG et al.; Acyl carrier protein (ACP) was purified from Euglena gracilis variety bacillaris in yields of about 1 mg/100 g (wet wt) of cells . Antibodies against the purified protein were raised in hens and isolated from eggs . Antibodies raised against Euglena ACP inhibited the Euglena chloroplast nonaggregated fatty acid synthetase using either Euglena or Escherichia coli ACP as a substrate . Comparisons with other ACPs included the following items: biologic activity, acidic pI, size, behavior in size exclusion media, and amino acid sequence of the N-terminal portion of the molecule.

Protein Expr Purif, 1991 Apr-Jun, 2(2-3), 151 - 7
Hyperexpression in Escherichia coli, purification, and characterization of the metallo-beta-lactamase of Bacillus cereus 5/B/6; Shaw RW et al.; We used site-directed mutagenesis to introduce both a NdeI restriction endonuclease site and an initiator codon at the junction of the leader and structural gene sequences of the metallo-beta-lactamase of Bacillus cereus 5/B/6 . This construct allowed us to clone just the beta-lactamase structural gene sequence into an Escherichia coli expression vector . E . coli cells were transformed with the recombinant plasmid, the B . cereus beta-lactamase was expressed, and these E . coli cells were disrupted by sonic oscillation . When the resultant suspensions were clarified by ultracentrifugation, the B . cereus beta-lactamase represented 15% of the total protein in the supernatant . Subsequent gel filtration and ion-exchange chromatography allowed the first reported purification to homogeneity of the B . cereus beta-lactamase from E . coli with an 87% recovery and an overall yield of 17 mg of enzyme per liter of cell culture . The electrophoretic mobilities of the enzyme expressed in and purified from E . coli and the enzyme purified directly from B . cereus were identical in both native and sodium dodecyl sulfate gel electrophoreses . As with the B . cereus enzyme, Km and Vmax (using cephalosporin C as substrate) for the enzyme purified from E . coli were 0.39 mM and 1333 units/mg protein, respectively . Likewise, the Co(II)-reconstituted enzyme purified from E . coli, which retained 29% of the activity of the Zn(II) enzyme, had electronic absorption spectra with maxima at 347, 551, 617, and 646 nm with extinction coefficients of 900, 250, 173, and 150 M-1 cm-1, respectively.

Indian J Lepr, 1991 Apr-Jun, 63(2), 203 - 8
Assessing the viability of Mycobacterium leprae by the fluorescein diacetate/ethidium bromide staining technique; Palomino JC et al.; In the present study we have evaluated the Fluorescein Diacetate/Ethidium Bromide (FDA/EB) staining technique to assess the viability of Mycobacterium leprae obtained from biopsies of leprosy patients under different periods of treatment . Bacillary suspensions were obtained from skin punch biopsies and stained with the FDA/EB solution . The average percentage of green cells seen which were deemed to be viable were: 67.2% of green cells in patients without previous treatment; 45.6% in patients with 1 to 6 months of treatment; 25.9% for patients with 7 to 12 months of treatment and 10.5% in patients with 13 to 24 months of treatment . All the patients studied were on multidrug therapy . The differences obtained in the percentages of green cells in the different groups of patients were statistically significant as determined by the Wilcoxon's test . The decrease in the percentage of green cells observed with increasing periods of treatment suggests that the FDA/EB technique correlates with the actual viability of M . leprae . The application of this technique in the routine procedures performed with Hansen's disease patients could be very useful for monitoring the effectiveness of treatment in leprosy patients.

J Bacteriol, 1991 Apr, 173(7), 2354 - 65
Characterization of the Cephalosporium acremonium pcbAB gene encoding alpha-aminoadipyl-cysteinyl-valine synthetase, a large multidomain peptide synthetase: linkage to the pcbC gene as a cluster of early cephalosporin biosynthetic genes and evidence of multiple functional domains; Gutierrez S et al.; A 24-kb region of Cephalosporium acremonium C10 DNA was cloned by hybridization with the pcbAB and pcbC genes of Penicillium chrysogenum . A 3.2-kb BamHI fragment of this region complemented the mutation in the structural pcbC gene of the C . acremonium N2 mutant, resulting in cephalosporin production . A functional alpha-aminoadipyl-cysteinyl-valine (ACV) synthetase was encoded by a 15.6-kb EcoRI-BamHI DNA fragment, as shown by complementation of an ACV synthetase-deficient mutant of P . chrysogenum . Two transcripts of 1.15 and 11.4 kb were found by Northern (RNA blot) hybridization with probes internal to the pcbC and pcbAB genes, respectively . An open reading frame of 11,136 bp was located upstream of the pcbC gene that matched the 11.4-kb transcript initiation and termination regions . It encoded a protein of 3,712 amino acids with a deduced Mr of 414,791 . The nucleotide sequence of the gene showed 62.9% similarity to the pcbAB gene encoding the ACV synthetase of P . chrysogenum; 54.9% of the amino acids were identical in both ACV synthetases . Three highly repetitive regions occur in the deduced amino acid sequence of C . acremonium ACV synthetase . Each is similar to the three repetitive domains in the deduced sequence of P . chrysogenum ACV synthetase and also to the amino acid sequence of gramicidin synthetase I and tyrocidine synthetase I of Bacillus brevis . These regions probably correspond to amino acid activating domains in the ACV synthetase protein . In addition, a thioesterase domain was present in the ACV synthetases of both fungi . A similarity has been found between the domains existing in multienzyme nonribosomal peptide synthetases and polyketide and fatty acid synthetases . The pcbAB gene is linked to the pcbC gene, forming a cluster of early cephalosporin-biosynthetic genes.

Wei Sheng Wu Xue Bao, 1991 Apr, 31(2), 94 - 9
{The study of Bacillus sphaericus Ts-1 protoplast-plasmid transformation of electroporation}; Wang Y et al.; This report gave the best conditions of Bacillus sphaericus Ts-1 protoplast-plasmid pHV33 electroporation . The highest transformation frequency and transformation efficiency induced by three pulse of 21 KV/cm and 10 microseconds duration applied at an interval of one sec., was 2.44 x 10(2) transformants/micrograms DNA and 3.16 x 10(-6) respectively . The saturated concentration of DNA absorbed by the protoplast was 5 micrograms DNA/10(9) cells/ml . By means of this method, pJB417, a recombinant mosquito larvicide clone, was introduced into B . subtilis 168M and B . sphaericus Ts-1 . The transformants of B . subtilis 168M with biocide activity were obtained, but the toxicity of B . sphaericus Ts-1 was not increased.

Antibiot Khimioter, 1991 Apr, 36(4), 5 - 8
{Isolation and identification of an antibiotic from culture 8-86}; Melikhov VA et al.; An antibiotic complex was isolated from culture 8-86 referred to Bacillus . The complex consisted of components 8-86A and 8-86B active against gram-negative organisms . By its physico-chemical properties such as IR and UV spectra, amino acid composition, specific rotation and fatty acid composition component 8-86B was shown to be close to polymyxin F.

J Mol Biol, 1991 Mar 20, 218(2), 465 - 75
Aromatic-aromatic interactions and protein stability . Investigation by double-mutant cycles; Serrano L et al.; The side-chains of phenylalanine and tyrosine residues in proteins are frequently found to be involved in pairwise interactions . These occur both within repeating elements of secondary structure and in tertiary and quaternary interactions . It has been suggested that they are important in protein folding and stability, and non-bonded potential energy calculations indicate that a typical aromatic-aromatic interaction has an energy of between -1 and -2 kcal/mol and contributes between -0.6 and -1.3 kcal/mol to protein stability . There is such an aromatic pair on the solvent-exposed face of the first alpha-helix of barnase, the small ribonuclease from Bacillus amyloliquefaciens . The edge of the aromatic ring of Tyr17 interacts with the face of that of Tyr13 . The two residues have been mutated both singly and pairwise to alanine, and their free energies of unfolding determined by denaturation with urea . Application of the double-mutant cycle analysis gives an interaction energy of -1.3 kcal/mol for the aromatic pair in the folded protein relative to solvation by water in the unfolded protein . This value is similar to that calculated from the change in surface-accessible area between the rings on the formation of the pair . Analysis of a further double-mutant cycle in which the Tyr residues are mutated to Phe indicates that the aromatic-aromatic interactions of Tyr/Tyr and Phe/Phe make identical contributions to protein stability . However, Tyr is preferred to Phe by 0.3(+/- 0.04) kcal/mol at the solvent-exposed face of the alpha-helix.

J Immunol, 1991 Mar 15, 146(6), 1987 - 95
Antigenicity and immunogenicity of a multiple peptidic construction of the Schistosoma mansoni Sm-28 GST antigen in rat, mouse, and monkey . 1 . Partial protection of Fischer rat after active immunization; Wolowczuk I et al.; Among the schistosome proteins characterized as vaccine candidates, an Ag of 28 kDa (Sm-28-GST) has received considerable attention . It was shown to be antigenic in humans and protective in mice, rats, hamsters, and baboons . Synthetic peptides derived from its sequence have been used to characterize the immune response to the molecule and one of these, comprising aminoacids 115-131 has been shown to incorporate both T and B cell recognition sites in a variety of experimental models . An octameric ("octopus") construction of the 115-131 peptide has been synthesized and its antigenicity and immunogenicity have been examined . The octopus construct is immunogenic in rats, mice and baboons in the presence of CFA (for rodents) and Bacille-Calmette-Guerin vaccine (for primates) as adjuvants . This clearly indicates that the construction allowed the conservation of the immune sites of the cognate protein . Moreover, anti-octopus sera from immunized Fischer rats were able to mediate platelet-, macrophage-, and eosinophil-dependent cytotoxicity toward schistosomula . Rats immunized with the 115-131 octopus were partially protected against a challenge infection with Schistosoma mansoni cercariae and this was paralleled by an increased level of IgG and more importantly, of IgE Sm-28-GST-specific antibodies.

Eur J Biochem, 1991 Mar 14, 196(2), 493 - 7
The ATPase of Bacillus alcalophilus . Reconstitution of energy-transducing functions; Hoffmann A et al.; The purified ATPase of Bacillus alcalophilus (F1F0) was reconstituted into proteoliposomes by gradual removal of the detergent Triton X-100 with Amberlite XAD-2 . The reconstitution was apparently highly asymmetric with nearly 100% of the F1 portion of the ATPase becoming oriented to the outside . Similar to results obtained with the soluble enzyme, the membrane-bound ATPase required Mg2+ and methanol for maximum activity . With Ca2+ or Mg2+ without methanol, 25% and 1%, respectively, of the maximum activity were observed . The ATPase was unable to pump Na+ ions but catalyzed the translocation of protons into the reconstituted proteoliposomes . Optimum proton translocation required the presence of Mg2+, not Ca2+, as divalent metal ion . The proton pump was inhibited by dicyclohexylcarbodiimide, venturicidin and NaN3 . On incubation of the reconstituted ATPase with {14C}dicyclohexylcarbodiimide, subunit c of the enzyme complex became specifically labeled . The proteoliposomes catalyzed the Mg2(+)-dependent incorporation of {32P}phosphate into ATP by ATP/{32P}phosphate exchange . This exchange was little affected by monensin, but was completely abolished by the uncoupler carbonyl cyanide m-chlorophenylhydrazone . Protons and not Na+ are thus the coupling ions of the ATPase of B . alcalophilus.

Am J Clin Pathol, 1991 Mar, 95(3), 418 - 23
Issues in cerebrospinal fluid management . Acid-fast bacillus smear and culture; Albright RE Jr et al.; Meningeal tuberculosis is an uncommon disease in the United States with an annual incidence of fewer than 200 cases . This study evaluates three approaches to improving the use of the cerebrospinal (CSF) acid-fast bacillus (AFB) smear and culture procedure: (1) education alone; (2) optional screening by which physicians can select to have the AFB analysis stopped if the initial CSF findings are unremarkable; and (3) mandatory screening before the performance of all CSF AFB analyses . With education alone, the CSF AFB culture rate decreased from 20.6% of all CSF acquisitions to 15.7% (P less than 0.001); however, the effect may have been related to a decrease in all types of AFB testing . Optional screening had no impact on the AFB testing rate . Mandatory screening significantly decreased the CSF AFB rate to 6.7% (P less than 0.001), unrelated to changes in other types of AFB testing . Laboratories that employ mandatory screening should report the screening results immediately and have a mechanism whereby physicians can bypass the screen, providing CSF AFB analysis on unremarkable fluid from high-risk patients.

Cell, 1991 Mar 8, 64(5), 1017 - 23
Barnase toxin: a new chimeric toxin composed of pseudomonas exotoxin A and barnase; Prior TI et al.; We have constructed a chimeric toxin composed of Pseudomonas exotoxin A (PE) and the extracellular ribonuclease of Bacillus amyloliquefaciens, barnase . The chimeric protein, termed PE-Bar, reacted with both anti-PE and anti-barnase antisera and had both ADP ribosylation and ribonuclease activities . The chimeric toxin was cytotoxic to the murine fibroblast cell line L929 and to a murine hybridoma resistant to PE . A mutant form of PE-Bar lacking ADP-ribosylating activity was still cytotoxic to L929 cells . Because treatment of cells prelabeld with {3H}uridine resulted in a decrease in their RNA content, we conclude that this cytotoxic effect was due to the ribonuclease activity of barnase molecules that had been translocated to the cytosol . It is now possible to construct chimeric toxins with two or more enzymatic activities that can be delivered to the cytosol of the target cells.

Immunol Lett, 1991 Mar, 27(3), 191 - 7
Cellular cytotoxicity of human natural killer cells and lymphokine-activated killer cells against bladder carcinoma cell lines; Wang MH et al.; The cytotoxicity of natural killer (NK) and lymphokine activated killer (LAK) cells against two human bladder tumor cell lines (BT-A and BT-B) was investigated using a fluorometric assay by labeling tumor cell DNA with Hoechst dye No . 33342 . Our results demonstrate that BT-A and BT-B cells have low sensitivity to the cytotoxic activity of mononuclear cells (MNC) and NK cells . Cytotoxicity of MNC or NK cells against both tumor cell lines is enhanced during co-culture of the effector cells with the target cells, which suggests that BT-A and BT-B cells provide the signals which could activate MNC to exert cytotoxicity . In contrast to NK cells, IL-2-generated LAK cells showed profound cytotoxicity to BT-A and BT-B within 24 h . In addition to cellular cytotoxicity to bladder tumor cells, we also tested the effect of recombinant interleukin 1 beta (rIL-1 beta), recombinant tumor necrosis factor (rTNF), and the supernatants of co-culture of MNC or LAK cells with bladder tumor cells . The results show no cytotoxic or growth-promoting activity of rIL-1, rTNF, or the crude culture supernatants on bladder tumor cells . We found that LAK cells, but not macrophages or NK cells, may play a major role in cellular cytotoxicity against the two bladder tumor cell lines tested . From this finding we conclude that activation of LAK cells may be one important mechanism induced by adjuvant bacillus Calmette-Guerin (BCG) therapy leading to effective prevention of urothelial bladder carcinoma reappearance.

J Med Entomol, 1991 Mar, 28(2), 219 - 22
Spiroplasma (Mollicutes: Spiroplasmataceae) pathogenic for Aedes aegypti and Anopheles stephensi (Diptera: Culicidae); Humphery-Smith I et al.; Intrathoracic inoculation with the mosquito spiroplasma, Spiroplasma taiwanense Abalain-Colloc et al., was found to reduce significantly the survival of adult male and female Aedes aegypti (L.) and Anopheles stephensi Liston . This spiroplasma also reduced significantly the flight capacity of adult female Ae . aegypti 5-8 d after inoculation and adult female An . stephensi 4 d after inoculation . Adult female An . stephensi were incapable of flight 5 d after inoculation . As such, S . taiwanense joins Bacillus thuringiensis and B . sphaericus as bacteria known to be pathogenic for mosquito vectors.

Pharm Res, 1991 Mar, 8(3), 341 - 4
Production of 5-15 microns diameter alginate-polylysine microcapsules by an air-atomization technique; Kwok KK et al.; A novel method of preparing small-sized microcapsules using a Turbotak air-atomizer is reported . Alginate-polylysine microcapsules containing Bacillus Calmette Guerin vaccine have been prepared by an adaptation of the method of Lim (1) which allows the manufacture of small-sized microcapsules . A Turbotak is used to spray sodium alginate solution into calcium chloride solution to form temporary calcium alginate microgel capsules . These temporary microgel droplets are subsequently cross-linked with polylysine to form permanent membranes . Microcapules in the size range of 5-15 microns have been produced which can be compared to an average diameter of greater than or equal to 300 microns obtained by the method reported by Lim . The microcapsule size is dependent on the conditions of operation of the Turbotak and the concentration of the sodium alginate solution . Particles within the size range 5-15 microns can be reproducibly manufactured using the conditions of operation reported here . Other size ranges below the minimum of 300 microns reported by Lim are also feasible using this technique.

J Am Mosq Control Assoc, 1991 Mar, 7(1), 56 - 62
Microbial larvicides for the control of nuisance aquatic midges (Diptera: Chironomidae) inhabiting mesocosms and man-made lakes in California; Rodcharoen J et al.; Bacillus thuringiensis var . israelensis (B.t.i.) and B . sphaericus were evaluated for chironomid larvicidal activity in freshwater mesocosms . Of two B.t.i . formulations, the technical powder ABG-6164 provided excellent control of chironomines (94%) at the rate of 11.2 kg/ha whereas the liquid concentrate, Vectobac 6 AS, achieved only moderate control (57%) at the rate of 22.4 kg/ha . In contrast, similar rates of B . sphaericus products, ABG-6184 technical powder and BSP-2 flowable concentrate, produced no significant reduction . Lake studies were initiated to determine which B.t.i . formulations were practical and effective in large-scale situations . Two sinking granular corn grit formulations worked with varying degrees of success but were deemed too bulky for extensive applications . B.t.i . technical powder preparations with high potency ratings of 5,000 and 12,430 ITU/mg gave excellent control of Chironomus spp . (100% and 87%) when used at the rates of 6.7 and 2.8 kg/ha, respectively . None of the treatments reduced larval populations of the tanypodines, Procladius and Tanypus.

J Gen Microbiol, 1991 Mar, 137 ( Pt 3), 667 - 77
Identification of four unique clones encoding 10 kDa proteins from Bacillus that cause phenotypic complementation of a phoA mutant strain of Escherichia coli; Lee JW et al.; A number of clones have been isolated from two Bacillus species which complement the PhoA- phenotype of Escherichia coli mutants under conditions that induce the expression of alkaline phosphatase (APase) . These clones were initially thought to carry XPases because the transformed host could hydrolyse a common APase substrate, XP (5-bromo-4-chloro-3-indolyl-phosphate) . The sequences of the open reading frames responsible for the phenotypic complementation showed no sequence similarity to ATPases of E . coli, human (bone-liver-kidney, intestinal or placental) or Bacillus . Therefore, these clones were designated as XPA (for X Phosphatase Activity) clones . Four of the clones encoded small (10 kDa), basic, hydrophobic proteins . Two of these, xpaB from B . subtilis 168 and xpaL2 from B . licheniformis MC14, shared 62% identity at both the DNA and the predicted amino acid sequence level . The fact that homologues from two Bacillus strains were cloned indicated that the screen was specific, but not for APase genes . It is clear that phenotypic complementation with cloned DNA from another genus does not ensure the identification of an APase gene . Possible mechanisms for the abnormal phenotypic complementation are discussed.

J Am Vet Med Assoc, 1991 Mar 1, 198(5), 862 - 3
Prolonged milk residue in two cows after subcutaneous injections of penicillin at an extra-label dose; Krainock RJ; After cesarian section was done on 2 Holstein cows, procaine penicillin G was injected SC at an extra-label-dose . The milk contained inhibitory residue for 21 days (case 1) and 10 days (case 2) after the final penicillin injection . The Bacillus stearothermophilus disk assay was used to confirm presence of an inhibitor . Analysis to specifically identify the inhibitor substance was not performed . Other sources of residues were examined and excluded in these cases . Procaine penicillin G was assumed to be the residue substance . Delayed drug clearance was attributed to depot formation following high-dose SC penicillin administration.

Gene, 1991 Mar 1, 99(1), 109 - 14
Promoter sequence analysis in Bacillus and Escherichia: construction of strong promoters in E . coli; Yamada M et al.; Many derivatives of the nprM promoter of Bacillus stearothermophilus and the strong early promoter, A3, of coliphage T3 were designed and chemically synthesized . These promoters consisted of some or all of the AT box, consensus sequence, tac promoter sequence, spacer, and lac operator . The promoter activities were assessed by their ability to express the cat gene in Escherichia coli . One of the derivatives of the A3 promoter, which contained the lac operator, was much stronger (about 3.5 times) than the tac promoter . The promoter activities in E . coli were considerably modified by the substitutions in the -43 (AT box) and -35 regions.

Eur J Immunol, 1991 Mar, 21(3), 793 - 7
Mycobacterial proliferation in macrophages is prevented by incubation with lymphocytes activated in vitro with a mycobacterial antigen complex; Beschin A et al.; Antigen A60 from Mycobacterium bovis bacillus Calmette Guerin was shown to trigger both humoral and cellular immune reactions . We explored the ability of A60 to block intracellular proliferation of phagocytosed mycobacteria with a model system involving peritoneal murine macrophages infected with Mycobacterium avium . Mixed lymphocytes from lymph nodes of mice inoculated with A60 hindered intracellular proliferation of this mycobacterium, owing to A60-specific cells, proliferation of which was induced in vitro in an antigen concentration-dependent manner . The lymphokines released by A60-stimulated T lymphocytes in vitro were identified as interleukin 2 (IL2) and interferon-gamma (IFN-gamma): their production showed a clear A60 dose dependence . When supernatants of such induced lymphocyte cultures were incubated with anti-IFN-gamma antibodies, macrophage activation was prevented, whereas anti-IL 2 immunoglobulin had little effect . Treatment of infected macrophages with recombinant IFN-gamma reduced intracellular proliferation of mycobacteria, while exogenous IL 2 and tumor necrosis factor were ineffective . Therefore, A60 elicits in vitro proliferation of T lymphocytes responding specifically to this antigen with production of IFN-gamma, which in turn activates macrophages and prevents multiplication of phagocytosed mycobacteria.

Biochem J, 1991 Mar 1, 274 ( Pt 2), 355 - 60
Purification and properties of adenosine 5'-phosphosulphate sulphotransferase from Euglena; Li JY et al.; Adenosine 5'-phosphosulphate sulphotransferase (APSST) was extracted from Euglena gracilis Klebs var . bacillaris mutant W10BSmL by freezing and thawing and was purified about 10,000-fold (to homogeneity) with 10.5% recovery by (NH4)2SO4 precipitation, Sephadex G-100 chromatography, Reactive Blue-agarose, Reactive Dye-agarose, DEAE-cellulose, preparative isoelectric focusing and non-inactivating SDS/PAGE . The active APSST, with a molecular mass of 102 kDa and multiple forms from pI 5.0 to 5.5, is a tetramer held together by covalent (probably disulphide) bonds . An apparent Km of the purified enzyme for adenosine 5'-phosphosulphate (APS) of 0.1 microM is obtained when dithiothreitol is used as the thiol . The enzyme is stimulated by Mg2+, Ca2+ or Ba2+, and uses almost any thiol; dithiothreitol and dithioerythritol give the highest activity . In the absence of APS, the enzyme is inactivated (and is rendered monomeric) by thiols but is protected from thiol inactivation by AMP, adenosine 5'-phosphoramidate (APA) or adenosine 5'-monosulphate (AMS), which also inhibit APSST activity somewhat . The enzyme resists inactivation by SDS in the absence of thiols; SDS stimulates APSST activity at low concentration, but high concentrations are inhibitory.

Am Rev Respir Dis, 1991 Mar, 143(3), 496 - 500
Cellular immunity in current active pulmonary tuberculosis; Andrade-Arzabe R et al.; A group of 10 patients with recently diagnosed pulmonary TB were studied and compared to 10 bacillus Calmette-Guerin (BCG) immunized healthy individuals . Cellular immune mechanisms were explored in vitro utilizing fresh and precultured peripheral blood mononuclear cells exposed to PHA, PPD, and recall antigens (SK/SD and CA) . Proliferative assays were also carried out in the presence of either each patient's serum (autologous serum) or cocultured with CD3(+)-depleted adherent cells . Serum measurements of soluble interleukin-2 (IL-2) receptor and synthesis of IL-2 generated by mononuclear cells stimulated with PPD and SK/SD were also performed . Patient sera were able to inhibit autologous as well as allogeneic cell responses, and a significant adherent cell suppressive effect was observed . As a whole the group of patients showed decreased blast transformation to PPD, preserved proliferative responses to other recall antigens, and a low PPD-induced generation of IL-2 . Furthermore, as possible evidence of preactivated T cells, these patients demonstrated high soluble IL-2 receptor serum levels . Early compromise of specific cell-mediated immunity, including IL-2 abnormalities, may be of significance in newly diagnosed pulmonary TB.

J Urol, 1991 Mar, 145(3), 498 - 501
Oral or intravesical bacillus Calmette-Guerin immunoprophylaxis in bladder carcinoma; D'Ancona CA et al.; A total of 71 patients with superficial transitional cell carcinoma underwent transurethral resection of bladder tumor . All patients had stage pTa or pT1 transitional cell carcinoma or carcinoma in situ without other concurrent malignancies . The patients were assigned to 3 treatment groups: control group--transurethral resection discontinued within the study, oral bacillus Calmette-Guerin (BCG) group--transurethral resection of bladder tumor plus BCG (Moreau) and intravesical BCG group--transurethral resection of bladder tumor plus BCG . Of 9 patients in the control group 8 (89%) experienced tumor recurrence during a mean followup of 20 months . Of the 28 patients in the oral BCG group 11 (39.3%) had recurrence during a mean followup of 36 months . Of the 34 patients in the intravesical group 6 (18%) had recurrence in a 24-month mean followup . The incidence of complications was higher in the intravesical (41.2%) than in the oral BCG group (28.5%) . These results show that intravesical BCG is a more effective immunotherapy; however, oral BCG can be used in patients who do not accept intravesical BCG administration.

Southeast Asian J Trop Med Public Health, 1991 Mar, 22(1), 108 - 12
Fermentation of a Malaysian Bacillus thuringiensis serotype H-14 isolate, a mosquito microbial control agent utilizing local wastes; Lee HL et al.; A screening program searching for indigenous microbial control agents of mosquitos in Malaysia is initiated since 1987 and to date at least 20 isolates of mosquitocidal Bacillus thuringiensis serotypes have been obtained . Preliminary field evaluation of several isolates indicated that they are highly effective in the control of medically important mosquito species . For operational purposes, there is an urgent need to produce this agent utilizing cheap and locally available wastes through fermentation biotechnology . Fermentation studies in shake-flasks containing standard nutrient broth and soya bean waste, respectively, indicate that it takes about 37 hours for a Malaysian isolate of B . thuringiensis serotype H-14 to mature . In the grated coconut waste, fishmeal and rice bran, the bacteria took 28 hours, 26 hours and 126 hours respectively to mature . The endotoxin was harvested from the standard nutrient broth at 55 hours and at 50 hours from soybean, grated coconut waste and fishmeal . The endotoxin could only be harvested 150 hours after inoculation from rice bran medium . However, no bacterial growth was detected in palm oil effluent . In terms of endotoxin and biomass production, fishmeal appears to be a suitable medium . Variations in the pH of the fermenting media were also noted.

FEMS Microbiol Lett, 1991 Mar 1, 62(2-3), 253 - 7
One-step purification procedure for UDP-N-acetylmuramyl-peptide murein precursors from Bacillus cereus; Kohlrausch U et al.; A method is described for the rapid isolation of the activated murein precursors UDP-N-acetyl-muramyl-pentapeptide (UDP-MurNAc-pentapeptide) and UDP-MurNAc-tripeptide from Bacillus cereus . After accumulation of the precursors by inhibition of murein synthesis either in the presence of vancomycin (for the pentapeptide precursor) or D-cycloserine (for the tripeptide precursor) the cells were extracted with boiling water . Prior to high pressure liquid chromatography the material was freed from acid precipitable material . UDP-MurNAc-penta- and tripeptide were separated from other components by reversed-phase HPLC on Hypersil ODS using isocratic elution conditions with sodium phosphate buffer . The precursors were obtained with at least 98% purity and a yield of about 50 mumol from a 10-l culture of B . cereus.

Thorax, 1991 Mar, 46(3), 220 - 1
Successful treatment of Bacillus cereus infection with ciprofloxacin; Gascoigne AD et al.; Bacillus cereus is rarely a pulmonary pathogen but may cause pneumonia in immunocompromised patients . A patient with bronchiectasis and no recognisable immunodeficiency had this organism isolated during two infective exacerbations, once from respiratory secretions and once by blood culture . Ciprofloxacin treatment was effective on both occasions.

J Bacteriol, 1991 Mar, 173(5), 1748 - 56
Effect of a 20-kilodalton protein from Bacillus thuringiensis subsp . israelensis on production of the CytA protein by Escherichia coli; Visick JE et al.; CytA, a 27-kDa cytolytic crystal protein of Bacillus thuringiensis subsp . israelensis, is produced only at very low levels by recombinant Escherichia coli cells unless a 20-kDa B . thuringiensis subsp . israelensis protein is also present (K . M . McLean and H . R . Whiteley, J . Bacteriol . 169:1017-1023, 1987; L . F . Adams, J . E . Visick, and H . R . Whiteley, J . Bacteriol . 171:521-530, 1989) . However, the data reported here demonstrate that the 20-kDa protein is not required for high-level CytA production in E . coli strains carrying mutations in rpoH, groEL, or dnaK, all of which affect the proteolytic ability of the cells . The 20-kDa protein also increases the amount of CryIVD (another B . thuringiensis subsp . israelensis crystal protein) and LacZX90 (a mutant of beta-galactosidase) made by E . coli . The latter phenomenon is attributable to an increase in the half-life of LacZX90, suggesting that the 20-kDa protein may stabilize this protein . The effect of the 20-kDa protein was also examined in vitro and in a T7 RNA polymerase expression system, and the possible significance of these results for the timing of proteolysis and of 20-kDa protein activity is discussed . Finally, the ability of a single antibody to coimmunoprecipitate CytA and the 20-kDa protein from E . coli extracts provides evidence for a protein-protein interaction that may be related to the mechanism of action of the 20-kDa protein.

Cesk Gynekol, 1991 Mar, 56(2), 93 - 6
{Problems of epidemiology and diagnosis in the vaginal area . II . Measurement of vaginal pH values and its relation to vaginal diseases}; Unzeitig V et al.; The authors divided a group of 600 women into 7 sub-groups based on the results of microscopic examination of native preparations, and using indicator papers of MERCK Co . (art . no . 9542) they assessed values of the vaginal pH . The highest values were recorded in a group of women with suppurative bacterial inflammations (mean pH 5.68) and the lowest ones in women with the finding of fungi (mean pH 5.0), cocobacillary flora without leucocytes (mean pII 4.39) and the normal physiological finding of Dorderlein's bacillus (mean pH 4.15) . Values higher than 4.8 rule out the possibility of a normal finding in the vagina, lower values, however, do not rule it out entirely . Assessment of pH values in the vagina should be considered part of routine examinations of the vaginal environment.

Pediatr Dermatol, 1991 Mar, 8(1), 21 - 4
Childhood leprosy in northern India; Kaur I et al.; During a period of eight years, 132 new leprosy cases were detected in children ages 3 to 19 years . Borderline tuberculoid leprosy was present in 59%, tuberculoid in 7.6%, and indeterminate type in 3.8% patients . Single skin lesions were seen in a significant number (43.9%) of patients . Bacillus-positive disease was detected more often (17.4%) than in adults . A high frequency (66.6%) of nerve involvement was also detected . Deformities were uncommon . Males were more often affected than females, especially in the ages 10 to 14 and 15 to 19 years . A history of contact was available in only 19.7% patients, and the contact was intrafamilial in 84.6%.

Bull Int Union Tuberc Lung Dis, 1991 Mar, 66(1), 33 - 6
Government intervention programs in HIV/tuberculous infection . Outline of guidelines for national tuberculosis control programs in view of the HIV epidemic; Kochi A; Tuberculosis is one of the most widespread infections known in the world . WHO estimates that in 1990, 1.7 billion people, or one third of the world population, are or have been infected with the tubercle bacillus . Fortunately, few of those infected develop active forms of the disease but it is estimated that in 1990, there will be 8 million new cases and 2.9 million deaths from tuberculosis in the world . This already alarming situation of the tuberculosis problem is getting worse, mainly due to the AIDS epidemic . A basic understanding of tuberculosis/HIV epidemiology is necessary and priority actions are to be strongly recommended for application in government intervention programs . They are specified in the present article.

Indian J Med Res, 1991 Mar, 93, 111 - 4
Isolation of mosquito-pathogenic Bacillus sphaericus & B . thuringiensis from the root surface of hydrophytes; Manonmani AM et al.; Attempts were made to isolate B . sphaericus and B . thuringiensis active against mosquito larvae from the root surface of hydrophytes . Out of 139 samples processed, 86 B . sphaericus and 23 B . thuringiensis isolates were obtained . Sixty two of the B . sphaericus isolates belonged to the serotype H5a5b, 2 to H6 and 22 isolates did not agglutinate with any of the 6 antisera tested . Twenty of the B . thuringiensis isolates belonged to the H14 serotype, 1 each to the H10 and H17 serotype(s) and 1 to an unknown serotype . Fifty nine of the B . sphaericus and 20 of the B . thuringiensis isolates fall under highly toxic category with the LC50 dose of 1-50 ng/ml for Culex quinquefasciatus third instar larvae.

J Invertebr Pathol, 1991 Mar, 57(2), 149 - 58
Cloning and expression of Bacillus thuringiensis israelensis delta-endotoxin DNA in B . sphaericus; Bar E et al.; Bacillus thuringiensis israelensis delta-endotoxin genes were cloned into Bacillus sphaericus 2362, producing stable transformants reacting with antibody to the 28- and 65-kDa B . thuringiensis israelensis crystal proteins and approximately 10 times more toxic to Aedes mosquito larvae than the original host strain . The LC50 after 48 hr of exposure of Aedes larvae to the most active transformed clone was 0.19 microgram/ml, compared with an LC50 of 1.9 microgram/ml for B . sphaericus 2362 and less than 0.1 microgram/ml for B . thuringiensis israelensis . The cloning vector, plasmid pPL603E, was also effective in transforming B . subtilis 1E20 with B . thuringiensis israelensis DNA, producing highly toxic clones with less stable gene expression than the clones of B . sphaericus.

J Exp Med, 1991 Mar 1, 173(3), 751 - 4
Listeria monocytogenes mutants lacking phosphatidylinositol-specific phospholipase C are avirulent; Camilli A et al.; A number of bacterial species secrete phosphatidylinositol-specific phospholipase C (PI-PLC) . In this report, we show that the facultative intracellular bacterial pathogen, Listeria monocytogenes, contains a gene, plcA, predicting a polypeptide with 31% amino acid identity to a Bacillus thuringiensis PI-PLC . Accordingly, L . monocytogenes secretes PI-PLC activity, while a mutant with a transposon insertion in plcA lacks detectable PI-PLC activity . In addition, expression of plcA in B . subtilis resulted in secretion of PI-PLC activity . The L . monocytogenes PI-PLC-defective mutant was three logs less virulent for mice and failed to grow in host tissues . The mutant was also defective for in vitro growth in mouse peritoneal macrophages . These results strongly suggest that PI-PLC is an essential determinant of L . monocytogenes pathogenesis . Whether the PI-PLC acts on a bacterial or host substrate remains to be determined.

J Neurochem, 1991 Mar, 56(3), 782 - 8
Purification and characterization of neuron-specific surface antigen defined by monoclonal antibody BM88; Patsavoudi E et al.; Monoclonal antibody BM88 recognizes a neurospecific surface antigen in the CNS and the PNS . In the present study, the antigen recognized by BM88 was immunopurified from pig brain and shown to be a 22-kDa polypeptide by reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Under nonreducing conditions a protein of 40 kDa was obtained, a result indicating that the antigen is composed of two polypeptide chains of equal molecular weight linked by disulfide bridges . Gel filtration of the purified antigen in the presence of Emulphogene suggested that it may be either a monomeric or a dimeric protein . However, in the presence of Triton X-100 a monomeric structure was implied . N-Glycanase digestion indicated that the protein is probably not glycosylated . The purified antigen was characterized as an integral membrane protein by hydrophobic chromatography and phase-separation experiments with Triton X-114 . The antigen, or at least the antibody binding region of the molecule, is very susceptible to protease attack, as judged by protease digestion experiments on brain membranes . By using very low concentrations of papain combined with short incubation times, the antigen was converted to a 16.3-kDa membrane-associated polypeptide as assessed by immunoblotting . This polypeptide contained the BM88 binding epitope . Soluble BM88 immunoreactive polypeptides were not obtained . Bacillus cereus phospholipase C was also unable to solubilize the antigen from the membrane . Our results suggest that the molecule, possessing at least one small extramembranous domain, is attached to the membrane via a polypeptide chain.

J Am Mosq Control Assoc, 1991 Mar, 7(1), 24 - 9
Small-scale field trials of Bacillus sphaericus (strain 2362) formulations against Mansonia mosquitoes in Malaysia; Yap HH et al.; Five formulations of Bacillus sphaericus (strain 2362) including aqueous suspension BSP 1, BSP 2, technical powder ABG 6184, corncob granules ABG 6185 (potencies 2 x 10(10), 2 x 10(7), 9.5 x 10(10), 5 x 10(10), 5 x 10(10) spore/g, respectively) and wettable powder ABG 6232 (1,000 BS ITU/mg) were tested against laboratory-cultured late third/early fourth instar larvae of Mansonia uniformis in floating screened cages in small plots at swampy ditches on Penang Island, Malaysia . Mean dosage/response values at 90% mortality levels were 6.93, 95.32, 1.45, 11.92 and 2.86 liters or kg per ha, respectively, for the formulations tested . There were practically no residual effects for the formulations tested with larvae introduced at 48, 96, and 168 h post-treatment . In trials of BSP 1, ABG 6184 and ABG 6185 (1 liter or 1 kg per ha) against immature Mansonia spp . in impounded paddy field ditches, improved efficacy and residual effects were obtained with mean reductions of 93.1, 91.9 and 80.4% at days 3, 7 and 14 posttreatment, respectively.

FEMS Microbiol Lett, 1991 Mar 1, 62(2-3), 293 - 6
Uptake of inorganic pyrophosphate by Bacillus megaterium; Villiotte PJ et al.; Cells of Bacillus megaterium take up inorganic pyrophosphate, employing a saturable carrier which is sensitive to sulfhydryl reagents, orthophosphate, and arsenate . Uptake is stimulated by proton ionophores, including CCCP and nigericin, indicating that proton cotransport can lead to an opposing gradient . Inhibitor sensitivity, as well a relatively high Km for inorganic pyrophosphate render it likely that uptake is mediated by an orthophosphate transport system.

J Biol Chem, 1991 Feb 25, 266(6), 3955 - 60
Purification and characterization of an activator protein for methanol dehydrogenase from thermotolerant Bacillus spp; Arfman N et al.; All thermotolerant methanol-utilizing Bacillus spp . investigated by us possess a NAD-dependent methanol dehydrogenase (MDH) activity which is stimulated by a protein present in the soluble fraction of Bacillus sp . C1 cells . This activator protein was purified to homogeneity from Bacillus sp . C1 cells grown at a low dilution rate in a methanol-limited chemostat culture . The native activator protein (Mr = 50,000) is a dimer of Mr = 27,000 subunits . The N-terminal amino acid sequence revealed no significant similarity with any published sequences . Stimulation of MDH activity by the activator protein required the presence of Mg2+ ions . Plots of specific MDH activity versus activator protein concentration revealed Michaelis-Menten type kinetics . In the presence of activator protein, MDH displayed biphasic kinetics (v versus substrate concentration) toward C1-C4 primary alcohols and NAD . The data suggest that in the presence of activator protein plus Mg2+ ions, MDH possesses a high affinity active site for alcohols and NAD, in addition to an activator- and Mg2(+)-independent low affinity active site . The activation mechanism remains to be elucidated.

J Mol Biol, 1991 Feb 20, 217(4), 731 - 6
Accurate measurements of coupling constants from two-dimensional nuclear magnetic resonance spectra of proteins and determination of phi-angles; Ludvigsen S et al.; A new and simple method to measure 3JHNH alpha coupling constants of proteins by adding and subtracting traces from corresponding two-dimensional nuclear Overhauser enhanced spectroscopy and two-dimensional correlated spectroscopy cross peaks after scaling is proposed . The optimal scaling for the addition and the subtraction of the two traces is obtained by minimizing an error function . The method was proven to give accurate and precise measurements of coupling constants when tested with a series of simulated spectra . The accuracy of the method was better than 0.1 Hz for all test cases including the limiting case of J = 2.0 Hz and line-width = 11.0 Hz . The accuracy of the method was better than 0.1 Hz for all test cases including The 3JHNH alpha coupling constants were measured in two-dimensional nuclear magnetic resonance spectra of the two proteins barley serine proteinase inhibitor (CI-2) and the bacterial ribonuclease (barnase) of Bacillus amyloliquefaciens . The experimentally measured coupling constants were used to calculate the constants in a Karplus equation to be: 3JHNH alpha = 6.7 cos2(phi-60) -1.3 cos(phi-60) +1.5 . These constants are in good accordance with those obtained for basic pancreatic trypsin inhibitor (BPTI) . In addition, special emphasis is given to the measurements of positive phi-angles, and to the contribution of molecular dynamics on the apparent coupling constants.

J Mol Biol, 1991 Feb 20, 217(4), 737 - 50
Structure of cyclodextrin glycosyltransferase refined at 2.0 A resolution; Klein C et al.; The previously reported structural model of cyclodextrin glycosyltransferase (EC 2.4.1.19) from Bacillus circulans has been improved . For this purpose the known sequence was built into an electron density map established by multiple isomorphous replacement and subsequent solvent-flattening at 2.5 A resolution . The resulting model was refined at 2.0 A resolution using a simulated annealing refinement method . Based on 70,171 independent reflections in the range 7.0 to 2.0 A resolution, a final R-factor of 17.6% was obtained with a model obeying standard geometry within 0.013 A in bond lengths and 2.7 degrees in bond angles . The final model consists of all 684 amino acid residues, two calcium ions and 588 solvent molecules.

Biochim Biophys Acta, 1991 Feb 15, 1076(3), 364 - 8
Aminoacylase I is not a glycolipid-anchored ectoenzyme in pig kidney; Greenhough KJ et al.; Subcellular fractionation of pig kidney cortex revealed that aminoacylase I (EC 3.5.1.14, N-acyl-L-amino-acid aminohydrolase) is predominantly a soluble enzyme with only 0.5% of the total activity being recovered in the membrane fraction . The aminoacylase I activity associated with the membrane preparations displayed neither rapid release following incubation with phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis nor the distinctive differential pattern of detergent solubilization which was seen with glycosyl-phosphatidylinositol-anchored proteins (renal dipeptidase, alkaline phosphatase) . When fractionated by phase separation in Triton X-114, integral membrane proteins of kidney microvillar membranes partitioned predominantly (greater than 90%) into the detergent-rich phase . In contrast, only 3.7% of aminoacylase I activity associated with microvillar membranes partitioned into the detergent-rich phase . Aminoacylase I activity of pig kidney would therefore appear to be a hydrophilic protein in nature and is not, as suggested previously, a G-PI-anchored integral membrane protein.

Biochem Biophys Res Commun, 1991 Feb 14, 174(3), 1077 - 83
Induction of group II-like phospholipase A2 by lipopolysaccharide in the liver of BCG-primed rat; Inada M et al.; The specific activity of phospholipase A2 (PLA2) in the liver homogenate was elevated 1.7-fold in bacillus Calmette-Guerin (BCG)-treated rats, 1.6-fold in lipopolysaccharide (LPS)-treated rats, and 2.4-fold in BCG-infected rats treated with LPS, compared with that of control rats . These increased activities were almost completely inhibited by the antibody directed against rat splenic group II PLA2 (PLA2M) but not by anti-pancreatic PLA2 antibody . The results of immunoblot analysis confirmed that the PLA2 immunochemically related to the group II enzyme was induced by treatment with BCG and/or LPS . The anti-PLA2M antibody-inhibitable PLA2 activity per a single cell was elevated not only in nonparenchymal cell fraction but also in hepatocyte fraction, as in the case of whole liver . On the contrary, the PLA2 concentration and its specific activity did not change by the same treatment both in spleen homogenate and in isolated spleen cell fractions although a 3-fold increase in spleen mass occurred by BCG treatment . These results suggested that a tissue-specific mechanism of the PLA2 induction by these inflammatory mediators may operate in liver.

Eur J Biochem, 1991 Feb 14, 195(3), 631 - 5
Two structural domains as a general fold of the toxic fragment of the Bacillus thuringiensis delta-endotoxins; Convents D et al.; The unfolding by guanidine hydrochloride of the toxic fragment of a Bacillus thuringiensis toxin belonging to the CryIC class reveals a two-step denaturation under both acid and alkaline conditions . This demonstrates the existence of two structural domains as building blocks for this toxin . Protease digests performed on a CryIA(b) and CryIC B . thuringiensis toxin, under native and partially denatured conditions, confirm this conclusion . Whereas the native CryIC toxin is completely protease resistant, the CryIA(b) toxin, earlier described as consisting of two structural domains {Convents, D., Houssier, C., Lasters, I . & Lauwereys, M . (1990) J . Biol . Chem . 265, 1369-1375}, is cleaved by three proteases, resulting in at least two common fragments . This suggests that this toxin is built up of two globular units linked by a protease-susceptible linker . The detection of a stable intermediate along the denaturation curve allows us to study and compare the consecutive unfolding of the structural domains for both toxins . By addition of a protease, under conditions where such an unfolding intermediate exists, a single denaturation phase can be assigned to a specific part of the protein . These experiments lead to the conclusion that the domain whose stability is highly dependent on pH corresponds to the N-terminal half of both toxins.

FEBS Lett, 1991 Feb 11, 279(1), 123 - 31
Modulation of protease specificity by a change in the enzyme microenvironment . Selectivity modification on a model substrate, purified soluble proteins and gluten; Hertmanni P et al.; Subtilisin BPN' activity on a synthetic substrate is found to decrease with the concentration of soluble additives such as sugars and polyols, the catalytic efficiency of the enzyme being related to the water activity in the reaction medium . Limited hydrolysis of B chain of insulin is followed and the cleavage priority determined . When carried out in glycerol-containing medium, both enzyme catalytic behaviour and specificity are perturbed; a different cleavage order and a selectivity restriction are observed . The experiments were generalised to purified proteins and to an insoluble protein complex . The hydrolysis kinetics of purified gliadins by pepsin and of gluten by a Bacillus neutral protease are modulated in presence of water activity depressors . Glycerol is able to increase both pepsin efficiency and gluten protein solubility . The hydrolysis order is affected by water-structuring molecules in the enzyme microenvironment and new peptides appear whatever the size and initial solubility of the substrate.

Chem Pharm Bull (Tokyo), 1991 Feb, 39(2), 417 - 20
Post-translational processing of tumor necrosis factor production; Kitahara-Tanabe N et al.; To elucidate the mechanism of tumor necrosis factor (TNF) production, we analyzed proteins produced in macrophages sharing the epitope of TNF according to the priming and triggering of TNF production . Rabbit alveolar macrophages primed with Bacillus Calmette-Guerin (BCG) were isolated and cultured in vitro with 35S-methionine, and the proteins produced were analyzed using anti-rabbit TNF monoclonal antibody . Primed with BCG, alveolar macrophages synthesized two proteins with molecular sizes of 50 and 17 kilodaltons (kDa) (p50 and p17) sharing the same epitope with mature TNF within the cells . These two proteins were released into the medium where other proteins were detected without TNF-activity . Cultured with lipopolysaccharide (LPS triggering), the primed alveolar macrophages released TNF-activity into the medium where p17 together with many larger proteins was detected by immunoprecipitation . In vitro translation of messenger ribonucleic acid (mRNA) from BCG-primed macrophages showed that primary TNF has a molecular size of 28 kDa (p28) . These results suggest that active TNF of p17 is secreted when triggered via post-translational processing of the precursor molecules synthesized through priming with BCG.

Curr Opin Immunol, 1991 Feb, 3(1), 91 - 6
Leprosy and cell-mediated immunity; Kaplan G et al.; The intradermal injection of the purified protein derivative of tuberculin into lepromatous leprosy patients leads to a local cell-mediated immune response and to the extensive destruction of Mycobacterium leprae . This local response also occurs after intradermal injection of recombinant human interleukin-2; when administered over an 8-day period interleukin-2 evokes a systemic cell-mediated immune response and a reduction in the bacillary burden.

Curr Opin Rheumatol, 1991 Feb, 3(1), 145 - 54
Rheumatic manifestations of neoplasia; Schwarzer AC et al.; Neoplasia in an important cause of rheumatic disease . The mechanisms are either direct infiltration of the bone and joint, or indirect infiltration with manifestations distant from the site of neoplastic involvement . Many of the reports reviewed in this article center on direct associations . In particular, there is a report on polymyositis and dermatomyositis and the link with neoplasia . Cases of reflex sympathetic dystrophy are also described in association with neoplasia . There is further discussion on the link of hairy cell leukemia and vasculitis . Other case reports highlight the multiple associations of musculoskeletal disease and neoplasia . These reports include patients with subcutaneous sarcoidosis, ankylosing spondylitis, polychondritis, and systemic sclerosis . Articular manifestations of benign pleural fibromas are described . The existence of these reports, however, does not constitute proof of a causal relationship and the possibility of chance occurrences of two conditions must be considered . Finally, various therapies for cancer are associated with rheumatologic manifestations . Intravesical bacillus Calmette-Guerin has been found to cause an inflammatory polyarthritis . This form of arthritis is similar to experimentally induced adjuvant arthritis in rats that follows immunization with Freund's adjuvant containing Mycobacterium tuberculosis.

An Esp Pediatr, 1991 Feb, 34(2), 129 - 31
{Contacts of children with tuberculous patients}; Castan Vidal ML et al.; One hundred and nine adults recently diagnosed of active pulmonary tuberculosis (all of them with smear positive sputum) were selected . Their household contacts under fifteen years of age were studied . 73.1 por 100 of the children were tuberculin-positive, and 33.1 por 100 of these "reactors" had developed a pulmonary tuberculosis themselves . The bacillary density in the sputum of the source case was correlated to percentage of infected and ill children among his contacts . Neonatal vaccination with BCG showed a protective effect against the illness in children under eight years of age.

Clin Invest Med, 1991 Feb, 14(1), 55 - 62
Influence of biological response modifiers of bacterial origin on disease progression in the MRL-lpr model of systemic lupus erythematosus; Hart DA et al.; Murine models of systemic lupus erythematosus exhibit some, but not all of the characteristics of human disease . Disease progression in the animal models is related to autoantibodies, genetics, and inflammatory processes . In this report the effects of two bacterial biological response modifiers (BRM) on disease progression in the MRL-lpr model were investigated . The two BRM tested were C . parvum and Bacillus-Calmette-Guerin (BCG), both of which are stimulators of the reticuloendothelial system and both of which have been shown by others to influence disease progression in NZB/W mice . Treatment of 10-week-old mice with C . parvum led to transient alterations in hepatosplenomegaly and plasma proteinase regulation, which then returned to control values . Treatment with BCG led to even more transient effects on the mice . Neither BRM appeared to impact on disease-associated alterations in autoantibody titres, hepatosplenomegaly, or elevations in plasma proteinase activity . Likewise, treatment of 17-week-old MRL-lpr mice with C . parvum did not influence disease progression as evidenced by survival, autoantibody production, or hepatosplenomegaly . Therefore, in contrast to the NZB/W strain, treatment of the MRL-lpr strain with these BRM does not appear to impact on disease progression . This difference may be due to the influence of the lpr accelerator gene in this model.

AIDS, 1991 Feb, 5(2), 195 - 9
Bacillus Calmette-Guérin immunization in infants born to HIV-1-seropositive mothers; Lallemant-Le Coeur S et al.; During the prospective follow-up of 64 babies at risk for perinatal HIV-1 infection because their mothers were seropositive, and of 130 control babies whose mothers were seronegative, we studied the occurrence of complications of bacillus Calmette-Guerin (BCG) immunization and its ability to induce cutaneous reactivity to tuberculin . Babies born both to HIV-1-positive and HIV-1-negative mothers received BCG immunization during their first month of life according to the Expanded Programme on Immunization (EPI) recommendations . Local and regional complications of BCG vaccine were looked for at 3, 6 and 9 months after inoculation . A tuberculin skin test was performed at 6 or 9 months of age . Most babies born to HIV-1-positive mothers were later classified as infected or uninfected according to their clinical condition and/or serological status at 18 months of age . The mean duration of the follow-up was 36 months (range 30-40 months) . No chronic or deep ulcerations at the site of injection or disseminated forms of BCG infection were observed . The frequency of BCG-related lymphadenitis in the group of HIV-1-infected children (24%) did not differ significantly from the group of uninfected children (19%; Fisher test: P = 0.73) . In contrast, the tuberculin skin test responses were positive less often in the group of HIV-1-infected children (33%) than in the uninfected group (83%; Fisher test: P = 0.007) . Because BCG vaccine appears to be safe--even when given to perinatally infected babies--continuation of the BCG immunization policies of the EPI is justified, especially in view of the growing incidence of tuberculosis as a complication of HIV infection.

Appl Environ Microbiol, 1991 Feb, 57(2), 349 - 58
Insecticidal toxins from Bacillus thuringiensis subsp . kenyae: gene cloning and characterization and comparison with B . thuringiensis subsp . kurstaki CryIA(c) toxins; Von Tersch MA et al.; Genes encoding insecticidal crystal proteins were cloned from three strains of Bacillus thuringiensis subsp . kenyae and two strains of B . thuringiensis subsp . kurstaki . Characterization of the B . thuringiensis subsp . kenyae toxin genes showed that they are most closely related to cryIA(c) from B . thuringiensis subsp . kurstaki . The cloned genes were introduced into Bacillus host strains, and the spectra of insecticidal activities of each Cry protein were determined for six pest lepidopteran insects . CryIA(c) proteins from B . thuringiensis subsp . kenyae are as active as CryIA(c) proteins from B . thuringiensis subsp . kurstaki against Trichoplusia ni, Lymantria dispar, Heliothis zea, and H . virescens but are significantly less active against Plutella xylostella and, in some cases, Ostrinia nubilalis . The sequence of a cryIA(c) gene from B . thuringiensis subsp . kenyae was determined (GenBank M35524) and compared with that of cryIA(c) from B . thuringiensis subsp . kurstaki . The two genes are more than 99% identical and show seven amino acid differences among the predicted sequences of 1,177 amino acids.

Gene, 1991 Feb 1, 98(1), 37 - 44
In vivo generation of hybrids between two Bacillus thuringiensis insect-toxin-encoding genes; Caramori T et al.; The parasporal crystal of Bacillus thuringiensis is composed of polypeptides highly toxic to a number of insect larvae . The structural genes (cryIA) encoding the Lepidoptera-specific toxin from different bacterial strains diverge primarily in a single hypervariable region, whereas the N-terminal and C-terminal parts of the proteins are highly conserved . In this report, we describe the generation of hybrid genes between two cryIA genes . Two truncated cryIA genes were cloned in a plasmid vector in such way as to have only the hypervariable region in common . The two truncated cryIA genes were separated by the tetracycline-resistance determinant (or part of it) . In vivo recombination between the hypervariable regions of the cryIA genes reconstituted an entire hybrid cryIA gene . Direct sequence analysis of 17 recombinant plasmids identified eleven different crossover regions which did not alter the reading frame and allowed the production of eight different hybrid proteins . The recombination events were independent from the RecA function of Escherichia coli . Some of the hybrid gene products were more specific in their insecticidal action and one had acquired a new biological activity.

Gene, 1991 Feb 1, 98(1), 141 - 5
Nucleotide sequence of the LYS2 gene of Saccharomyces cerevisiae: homology to Bacillus brevis tyrocidine synthetase 1; Morris ME et al.; The Saccharomyces cerevisiae LYS2 gene, which encodes alpha-aminoadipate reductase, an essential enzyme in the yeast lysine biosynthetic pathway, has been sequenced . A large open reading frame (ORF) has been identified which can specify a 1392-amino acid protein with a deduced Mr of 155,344 . A DNA database search using the translated LYS2 ORF as a probe has revealed significant aa sequence homology to the Bacillus brevis enzyme tyrocidine synthetase 1.

Mol Gen Genet, 1991 Feb, 225(2), 177 - 85
Hybrid Bacillus (1-3,1-4)-beta-glucanases: engineering thermostable enzymes by construction of hybrid genes; Olsen O et al.; Hybrid (1-3,1-4)-beta-glucanase genes were constructed by extension of overlapping segments of the (1-3,1-4)-beta-glucanase genes from Bacillus amyloliquefaciens and B . macerans generated by the polymerase chain reaction (PCR) . Four hybrid genes were expressed in Escherichia coli cells . The mature hybrid enzymes contain a 16, 36, 78, or 152 amino acid N-terminal sequence derived from B . amyloliquefaciens (1-3,1-4)-beta-glucanase followed by a C-terminal segment derived from B . macerans (1-3,1-4)-beta-glucanase . Biochemical characterization of parental and hybrid enzymes shows a significant increase in thermostability of three of the hybrid enzymes when exposed to an acidic environment thus combining two important enzyme characteristics within the same molecule . At pH 4.1, 85%-95% of the initial activity was retained after 1 h at 65 degrees C in contrast to 5% and 0% for the parental enzymes from B . amyloliquefaciens and B . macerans . After 60 min incubation at 70 degrees C, pH 6.0, the parental enzymes retained 5% or less of the initial activity whilst one of the hybrids still exhibited 90% of the initial activity . Of the parental enzymes B . macerans (1-3,1-4)-beta-glucanase had the lower specific activity while the hybrid enzymes exhibited specific activities that were 1.5- to 3-fold higher . These experimental results demonstrate that exchange of homologous gene segments from different species may be a useful technique for obtaining new and improved versions of biologically active proteins.

Eur J Immunol, 1991 Feb, 21(2), 431 - 7
Regulation of tumor necrosis factor (TNF) release by murine peritoneal macrophages: role of cell stimulation and specific phagocytic plasma membrane receptors; Stein M et al.; In spite of the physiologic and pathologic importance of tumor necrosis factor (TNF), the cellular factors that govern its release by macrophages (M phi) are poorly understood, in comparison with other secretory products . We have studied the role of M phi heterogeneity and of plasma membrane receptors in regulating TNF release in vitro . Resident and various exudate murine peritoneal M phi populations were challenged with lipopolysaccharide (LPS) or different phagocytic particles, and TNF release assayed by cytotoxicity for L-929 fibroblasts . Resident peritoneal M phi (RPM phi) released a small amount of TNF in response to LPS whereas thioglycollate-elicited M phi (TPM phi) released high levels of TNF (5000 U/3 x 10(5) M phi/ml) . M phi elicited by Bio-Gel polyacrylamide beads (BgPM phi), another nonspecific inflammatory stimulus, or early in the course of intraperitoneal Bacillus Calmette-Guerin infection, before recruited cells become immunologically activated, released tenfold less TNF after the same stimulus . By contrast, TNF release in response to various phagocytic triggers was similar (approximately 300-600 U/3 x 10(5) M phi/ml) in all M phi populations including RPM phi . The response by BgPM phi to LPS was enhanced by pre-treatment in vitro with interferon-gamma or thioglycollate broth . With respect to phagocytic receptor triggering we found that complement receptor type 3 (CR3) ligation or latex uptake did not mediate release of significant quantities of TNF (less than 48 U/3 x 10(5) M phi/ml) by any M phi, whereas ligation of the Fc receptor for IgG1/IgG2b subclasses or of receptors for zymosan particles sufficed, in the absence of ingestion, to induce release of circa 500 U/3 x 10(5) M phi/ml TNF by all M phi tested . Our studies show that M phi vary in respect to priming for TNF release and that heterogeneity should be related to a particular triggering stimulus . Furthermore, the capacity of some M phi populations to release unusually high levels of TNF depends on immune or nonspecific stimuli subsequent to the process of inflammatory recruitment.

Biochem J, 1991 Feb 1, 273 ( Pt 3), 503 - 10
Ragged N-termini and other variants of class A beta-lactamases analysed by chromatofocusing; Matagne A et al.; Four beta-lactamases excreted by Gram-positive bacteria exhibited microheterogeneity when analysed by chromatofocusing or ion-exchange chromatography . Ragged N-termini were in part responsible for the charge variants, but deamidation of an asparagine residue was also involved, at least for the Bacillus licheniformis enzyme . The activity of a contaminating proteinase could also be demonstrated in the case of Actinomadura R39 beta-lactamase . With that enzyme, proteolysis resulted in partial inactivation, but the inactivated fragments were easily separated from the active forms . With these, as with the other enzymes, the kinetic parameters of the major variants were identical with those of the mixture within the limits of experimental error, so that the catalytic properties of these enzymes can be determined with the 'heterogeneous' preparations.

Semin Oncol, 1991 Feb, 18(1 Suppl 1), 27 - 38
The role of immunotherapy in colorectal cancer; Wadler S; Despite landmark advances in the molecular biology and surgical adjuvant therapy of colorectal cancer in the past decade, this illness remains a significant health hazard . Conventional therapies, including surgery, radiation therapy, and chemotherapy, have had limited utility and have often resulted in unacceptable host toxicities . Therapies dependent on potentiation of the host immune response are attractive alternatives to conventional treatment because of their greater specificity and diminished toxicity . Furthermore, the efficacy of many of these therapies has been demonstrated in preclinical models, which is important given the relative refractoriness of colorectal cancer to chemotherapy . Both active specific therapies, such as tumor vaccines, and passive therapies, such as monoclonal antibodies, have been investigated . In early clinical trials, both have demonstrated modest activity . Nonspecific immune stimulation with agents such as bacillus Calmette-Guerin or methanol extraction residue appear to have little utility, but with the development of recombinant DNA technology, specific cytokines with more precisely identified targets have been synthesized in sufficient quantities for clinical trials . Finally, combinations of biologics or combinations of biologics with conventional therapies, such as fluorouracil (5-FU)/levamisole and 5-FU/interferon, have been investigated and appear to be useful in the treatment of surgically resected and advanced colorectal carcinoma, respectively . The utility of these therapies is hampered by a limited understanding of their precise mechanisms of action and optimal conditions for administration . Nevertheless, immunotherapy of colorectal carcinoma remains an important and promising area for further clinical investigation.

Proc Natl Acad Sci U S A, 1991 Feb 1, 88(3), 1054 - 8
Recombinant fusion protein identified by lepromatous sera mimics native Mycobacterium leprae in T-cell responses across the leprosy spectrum; Laal S et al.; Pooled polyvalent sera from lepromatous leprosy patients were used to screen a lambda gt11 recombinant DNA expression library of Mycobacterium leprae in order to identify the relevant antigens recognized by the human immune response . Of the 300,000 phages screened, 4 clones were identified that coded for fusion proteins of the same molecular mass . The fusion protein from clone LSR2 was tested for immunoreactivity in assays using peripheral blood cells and sera from 11 laboratory personnel and 105 patients across the leprosy spectrum . LSR2 protein appears to be predominantly a T-cell antigen . It evokes similar lymphoproliferative responses as the native bacillus both at the individual level and in the leprosy spectrum as a whole . Though only 50% of patient sera with anti-M . leprae antibodies reacted with the fusion protein, the pattern of reactivity in the antibody responses was also similar for the various clinical types . The coding regions of clones LSR1 and LSR2 are identical . They show no homology with sequences stored in data banks and encode a protein of 89 amino acids with a calculated molecular mass of approximately 10 kDa.

J Bacteriol, 1991 Feb, 173(3), 1353 - 6
Efficient transformation of Bacillus thuringiensis requires nonmethylated plasmid DNA; Macaluso A et al.; The transformation efficiency of Bacillus thuringiensis depends upon the source of plasmid DNA . DNA isolated from B . thuringiensis, Bacillus megaterium, or a Dam- Dcm- Escherichia coli strain efficiently transformed several B . thuringiensis strains, B . thuringiensis strains were grouped according to which B . thuringiensis backgrounds were suitable sources of DNA for transformation of other B . thuringiensis strains, suggesting that B . thuringiensis strains differ in DNA modification and restriction . Efficient transformation allowed the demonstration of developmental regulation of cloned crystal protein genes in B . thuringiensis.

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1991 Feb, 24(1), 30 - 5
Diagnostic value of measuring BCG sonicate antigen and anti-BCG antibodies in the cerebrospinal fluid and blood of patients with tuberculous meningitis; Lin HP et al.; Fourteen cerebrospinal fluid (CSF) and paired blood samples were obtained from patients with tuberculous meningitis, seven with positive culture and seven clinical/laboratory diagnosis . Another 14 paired specimens served as control, including 7 infectious meningitis and 7 non-inflammatory neurological diseases . Four groups were thus classified, including confirmed and suspected patients, and controls with infectious meningitis and neurological diseases . Measurements of Bacillus Calmette-Guerin (BCG) sonicate antigen and IgG antibody were performed using enzyme-linked immunosorbent assay . Generally, CSF BCG sonicate antigen level and anti-BCG sonicate antibody of patients with tuberculosis meningitis were higher than in control groups; greater antigen levels were found in confirmed patients than in non-inflammatory subjects (p less than 0.05), and in suspected patients than in infectious and non-inflammatory subjects (p less than 0.05 and p less than 0.01) . For serum anti-BCG sonicate levels, confirmed patients had higher antibody value than non-inflammatory subjects (p less than 0.05) . To conclude, detection of high levels of both BCG sonicate antigen and antibody in CSF and blood samples shows great value in the diagnosis of tuberculous meningitis . However, given the limited samples of the current research, more data are needed to elucidate the sensitivity and specificity of such tests.

Nippon Hinyokika Gakkai Zasshi, 1991 Feb, 82(2), 290 - 6
{Intravesical instillation of bacillus Calmette-Guerin for superficial bladder carcinoma: study on significance of additional maintenance instillations of bacillus Calmette-Guerin}; Yabusaki N et al.; The efficacy of maintenance bacillus Calmette-Guerin (BCG) instillations for superficial bladder tumors was studied by prospective randomized trial . From June 1985 to October 1988, 42 newly diagnosed patients with superficial bladder carcinoma (pTa or pT1) were treated by transurethral tumor resection and subsequent five daily instillations of mitomycin C . Then they were divided into non-maintenance group (22 patients) and maintenance group (20 patients) by randomization . The patients received six weekly instillations of 80 mg of BCG . Tokyo strain (Japan BCG manufacturing Co., Tokyo, Japan), suspended in 40 ml of physiological saline, and the patients in the maintenance group received four additional instillations of BCG every three months . We could not complete the six-week course of BCG instillations in three patients due to adverse effects (two in non-maintenance group and one in maintenance group) and we lost six patients from follow-up within one year (one in non-maintenance group and five in maintenance group) . The mean follow-up period of the remaining 33 patients was 28.1 months . Of these 33 patients, six patients had been found to have recurrent tumors, and the over-all three-year non-recurrence rate was 82% . Before employing BCG, when we used only mitomycin C after TUR-Bt, the three year non-recurrence rate was 58% . This indicates prophylactic effect of BCG instillations . The stage of the initial tumor of the six recurrent cases were all pT1b . The non-recurrence rate of the patients with pT1b tumor was significantly lower than that of the patients with pTa and pT1a tumor . However, multiplicity and grade of tumors did not affect the non-recurrence rate.(ABSTRACT TRUNCATED AT 250 WORDS)

J Gen Microbiol, 1991 Feb, 137 ( Pt 2), 399 - 404
Structural and stereospecific requirements for the nucleoside-triggered germination of Bacillus cereus spores; Senesi S et al.; A selection of adenosine analogues was tested for their ability to trigger germination of Bacillus cereus NCIB 8122 spores . The germination-inducing activity was governed by the structural properties of the sugar rather than the base moieties of the nucleosides . Among the sugar-modified analogues, only those containing a 2'-deoxy-D-ribose moiety promoted spore germination . Requirements for a specific molecular structure of the base were not clearly identified, although the highest activity was observed when substituents were inserted at position 6 of the purine ring . All the base-modified analogues, even those such as coformycin and 2'-deoxycoformycin with an expanded base ring, retained the germination-inducing activity of adenosine . However, of the two 2'-deoxycoformycin diastereoisomers characterized by an asymmetric carbon atom at position 8 of the homopurine ring, only the 8S-isomer induced germination, thus indicating that stereospecific configuration of the inducer, at least in the case of 2'-deoxycoformycin, appears to be essential for the initiation of spore germination . The differences in the germination-inducing activity of the various analogues tested were not affected significantly by spore activation at different temperatures, although the higher the activation temperature, the lower was the concentration of each analogue required for maximum germination.

Appl Environ Microbiol, 1991 Feb, 57(2), 480 - 5
Biolistic transformation of a procaryote, Bacillus megaterium; Shark KB et al.; We present a simple and rapid method for introducing exogenous DNA into a bacterium, Bacillus megaterium, utilizing the recently developed biolistic process . A suspension of B . megaterium was spread onto the surface of nonselective medium . Plasmid pUB110 DNA, which contains a gene that confers kanamycin resistance, was precipitated onto tungsten particles . Using a biolistic propulsion system, the coated particles were accelerated at high velocities into the B . megaterium recipient cells . Selection was done by use of an agar overlay containing 50 micrograms of kanamycin per ml . Antibiotic-resistant transformants were recovered from the medium interface after 72 h of incubation, and the recipient strain was shown to contain the delivered plasmid by agarose gel electrophoresis of isolated plasmid DNA . All strains of B . megaterium tested were successfully transformed by this method, although transformation efficiency varied among strains . Physical variables of the biolistic process and biological variables associated with the target cells were optimized, yielding greater than 10(4) transformants per treated plate . This is the first report of the biolistic transformation of a procaryote.

Genetika, 1991 Feb, 27(2), 238 - 46
{Inversion polymorphism in malaria mosquito Anopheles messeae . Part X . Resistance of larvae with different genotypes to toxins of crystal-forming bacteria Bacillus thuringiensis subsp . israelensis (serovar H14)}; Gordeev MI et al.; Insensibility of larvae with different chromosomal inversions to the toxins of Bacillus thuringiensis subsp . israelensis (Bti) was examined . It has been shown that larvae with inversion combinations XL0(XL1)2R0-3R0-3L0 had greater variability after treatment with Bti than larvae with inversions XL1(XL2)2R1-3R1-3L1(3L0) . The former inversion combinations were previously shown to dominate in the south of the species area and to be supported by K-selection . The latter inversion combinations form "northest" karyotypes and are supported by r-selection . It is obvious that genetic effects of treatments with Bti depend on the population's structure and the directions of natural selection . The decrease in the level of heterozygotes after treatment of larvae with Bti reflects destruction of the system of genetic homeostasis in the natural populations of A . messeae.

J Biochem (Tokyo), 1991 Feb, 109(2), 211 - 6
Purification and characterization of a novel thermostable lipase from Bacillus sp; Sugihara A et al.; A thermostable lipase from Bacillus sp . has been purified to homogeneity as judged by disc-PAGE, SDS-PAGE, and isoelectric focusing . The purification included ammonium sulfate fractionation, treatment with acrinol, and sequential column chromatographies on DEAE-Sephadex A-50, Toyopearl HW-55F, and Butyl Toyopearl 650M . The purified enzyme was found to be a monomeric protein with Mr of 22,000, and pI of 5.1 . The optimal pH at 30 degrees C, and optimal temperature at pH 5.6 were 5.5-7.2, and 60 degrees C, respectively, when olive oil was used as the substrate . The substrate specificity towards simple triglycerides was broad and 1- and 3-positioned ester bonds were hydrolyzed in preference to a 2-positioned ester bond . The addition of acetone to the assay mixture in the range of 0-60% (v/v) stimulated the enzyme remarkably, whereas n-hexane had an inhibitory effect.

Enferm Infecc Microbiol Clin, 1991 Feb, 9(2), 102 - 5
{Endocarditis caused cy Erysipelothrix rhusiopathiae . Study of 2 cases and review of the literature}; Azofra J et al.; Endocarditis produced by E . rhusiopathiae is a uncommon disease . Most of the infected persons (90%) work in environments with frequent exposure to E . rhusiopathiae (butchers, fisherman) . Although the clinical picture of endocarditis produced by E . rhusiopathiae is indistinguishable from other forms of subacute endocarditis, this infection has a mortality rate of 40% and a high morbidity . Microbiological diagnosis should consider the possibility of making a mistake considering that isolation of a gram-positive bacillus may represent contamination by an agent without clinical relevance . Treatment with penicillin G during 4 weeks is commonly sufficient to cure the disease.

Cell Mol Neurobiol, 1991 Feb, 11(1), 219 - 30
Rapid analysis of glycolipid anchors in amphiphilic dimers of acetylcholinesterases; Toutant JP et al.; 1 . We describe two simple procedures for the rapid identification of certain structural features of glycolipid anchors in acetylcholinesterases (AChEs) . 2 . Treatment with alkaline hydroxylamine (that cleaves ester-linked acyl chains but not ether-linked alkyl chains) converts molecules possessing a diacylglycerol, but not those with an alkylacylglycerol, into hydrophilic derivatives . AChEs in human and bovine erythrocytes possess an alkylacylglycerol (Roberts et al., J . Biol . Chem . 263:18766-18775, 1988; Biochem . Biophys . Res . Commun . 150:271-277, 1988) and are not converted to hydrophilic dimers by alkaline hydroxylamine . Amphiphilic dimers of AChE from Drosophila, from mouse erythrocytes, and from the human erythroleukaemia cell line K562 also resist the treatment with hydroxylamine and likely possess a terminal alkylacylglycerol . This indicates that the cellular pool of free glycolipids used as precursors of protein anchors is distinct from the pool of membrane phosphatidylinositols (which contain diacylglycerols) . 3 . Pretreatment with alkaline hydroxylamine is required to render the amphiphilic AChE from human erythrocytes susceptible to digestion by Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PI-PLC) (Toutant et al., Eur . J . Biochem . 180:503-508, 1989) . We show here that this is also the case for the AChE from mouse erythrocytes, which therefore likely possesses an additional acyl chain in the anchor that prevents the action of PI-PLC . 4 . In two sublines of K562 cells (48 and 243), we observed that AChE either was directly susceptible to PI-PLC (243) or required a prior deacylation by alkaline hydroxylamine (48) . This suggests that glycolipid anchors in AChE of K562-48 cells, but not those in AChE of K562-243 cells, contain the additional acylation demonstrated in AChE from human erythrocytes . These observations illustrate the cell specificity (and the lack of species-specificity) of the structure of glycolipid anchors.

Hindustan Antibiot Bull, 1991 Feb-Nov, 33(1-4), 19 - 25
Role of side chain moiety in the hydrolysis of penicillins by beta-lactamase; Ambedkar SS et al.; Various beta-lactam compounds and structurally related moieties were examined as substrates of beta-lactamase from Bacillus cereus 5/B NCTC 9946 . The enzyme was specific for penicillins and none of the cephalosporins were hydrolysed . Electronic environment of allylic carboxy group in dihydrothiazine ring restricts the acceptance of cephalosporins as substrates . The efficiency of hydrolysis of penicillins is dependent on dense resonating electronic environment of phenyl ring present in the side chain, flexibility of the side chain and the distance between the phenyl ring and carbonyl group in the side chain.

Immunol Invest, 1991 Feb, 20(1), 33 - 43
The effect of alpha 2 macroglobulin-proteinase complexes on macrophage Ia expression in vivo; DeStefano A et al.; Alpha-2-Macroglobulin (alpha 2M) is a major plasma proteinase inhibitor . It can also regulate the function of cells of the immune system, including macrophage expression of Ia antigens in tissue culture systems . The present work was done to assess the effect of alpha 2M-trypsin complexes (alpha 2M-t) on macrophage Ia expression in vivo . Bacillus Calmette-Guerin-infected mice were injected intraperitoneally with 100nM alpha 2M-t, phosphate buffered saline (PBS), or bovine serum albumin (BSA) in PBS . The peritoneal cells were harvested by lavage from 3 to 6 days after injection . Differential cell counts were performed, and macrophage Ia antigen expression determined by indirect immunofluorescence . Injection of either alpha 2M-t or BSA solutions tended to increase the number of total cells and lymphocytes harvested, without changing the number of macrophages harvested . alpha 2M-t injection reduced the proportion of macrophages which were Ia positive from 60 to 37% on day 3 after injection, and to 20% Ia positive on day 6 . The reduction in Ia positive macrophages was statistically significant when compared to either PBS or BSA injected groups . In summary, in vivo exposure to alpha 2M-t can alter macrophage function . alpha 2M-proteinase complexes formed during the course of coagulation or inflammation may play a physiologic role as regulators of the immune response.

Protein Expr Purif, 1991 Feb, 2(1), 51 - 8
High-level expression in Escherichia coli and rapid purification of phosphatidylinositol-specific phospholipase C from Bacillus cereus and Bacillus thuringiensis; Koke JA et al.; The construction of four vectors for high-level expression in Escherichia coli of the phosphatidylinositol-specific phospholipase C from Bacillus cereus or Bacillus thuringiensis is described . In all constructs the coding sequence for the mature phospholipase is precisely fused to the E . coli heat-stable enterotoxin II signal sequence for targeting of the protein to the periplasm . In one set of plasmids expression of the B . cereus or B . thuringiensis enzyme is under control of the E . coli alkaline phosphatase promoter, while in a second set of plasmids expression is under control of a lac-tac-tac triple tandem promoter . A simple and rapid procedure for complete purification of the phospholipase C overproduced in E . coli, involving isolation of the periplasmic proteins by osmotic shock followed by a single column chromatography step, is described . The largest quantity of purified enzyme, 40-60 mg per liter culture, is obtained with the plasmid expressing the B . cereus enzyme under control of the lac-tac-tac promoter . Lower quantities are obtained with the plasmids containing the alkaline phosphatase promoter (15-20 and 4-6 mg/liter for the B . cereus and B . thuringiensis enzymes, respectively) and with the plasmid expressing the B . thuringiensis phospholipase under control of the lac-tac-tac promoter (15-20 mg/liter) . A comparison of the functional properties of the recombinant phospholipases with the native enzymes isolated from B . cereus or B . thuringiensis culture supernatant shows that they are identical with respect to their catalytic functions, viz., cleavage of phosphatidylinositol and cleavage of the glycosyl-phosphatidylinositol membrane anchor of bovine erythrocyte acetylcholinesterase.

Mol Microbiol, 1991 Feb, 5(2), 367 - 72
Identification of phosphatidylinositol-specific phospholipase C activity in Listeria monocytogenes: a novel type of virulence factor?
Mengaud J, Braun-Breton C, Cossart P.
A phospholipase C which cleaves phosphatidylinositol and glycosylphosphatidylinositol (GPI) anchors was identified in Listeria monocytogenes . This 36 kDa protein is encoded by the gene plcA, and is homologous to the Bacillus cereus, Bacillus thuringiensis and eukaryotic phosphatidylinositol-specific phospholipases C (PI-PLC) . Expression of the plcA gene in Escherichia coli correlates with the appearance of PI-PLC activity in the cells . In Listeria monocytogenes, the activity is secreted to the culture medium . PI-PLC activity was only found in the two pathogenic species of the genus Listeria, namely L . monocytogenes and L . ivanovii . PI-PLC activity was lost and virulence decreased when the plcA gene was disrupted in the chromosome . This suggests that the PI-PLC of L . monocytogenes might be involved in virulence.

J Biotechnol, 1991 Feb, 17(2), 121 - 31
Production of 5-methyluridine by immobilized thermostable purine nucleoside phosphorylase and pyrimidine nucleoside phosphorylase from Bacillus stearothermophilus JTS 859; Hori N et al.; 5-Methyluridine was produced continuously from thymine and inosine by immobilized enzymes, which consisted of thermostable purine nucleoside phosphorylase and thermostable pyrimidine nucleoside phosphorylase obtained from Bacillus stearothermophilus JTS 859 . The process was carried out in a column reactor at 60 degrees C for 17 d without any bacterial contamination under non-aseptical conditions . Half-lives of the activity of the immobilized enzymes were 47 d and 4.5 d at 60 degrees C and 70 degrees C, respectively, although half-life of the crude enzyme was only 14 h at 70 degrees C.

Biochemistry, 1991 Jan 29, 30(4), 1028 - 36
Detection and characterization of intermediates in the folding of large proteins by the use of genetically inserted tryptophan probes; Smith CJ et al.; L-Lactate dehydrogenase from Bacillus stearothermophilus was rebuilt by using site-directed mutagenesis to produce an enzymically active, tryptophan-less enzyme by replacing all the wild-type tryptophans (80, 150, and 203) by tyrosines . Nine single tryptophan-containing active enzymes were constructed from this enzyme by genetically replacing one of the tyrosines 36, 85, 147, 190, 203, 237, 248, 279, or 285 by tryptophan . The equilibrium and the time-resolved tryptophan fluorescence intensity and anisotropy were used to report unfolding events in guanidine hydrochloride (GHCl) monitored from these nine defined positions . Three structural transitions, half complete at 0.55, 1.7, and 2.8 M GHCl, were identified and defined four folding intermediates, I (native), II (expanded monomer 1), III (expanded monomer 2), and IV (random coil), stable at 0, 1, 2.2, and 4 M GHCl, respectively . Intermediate II is a globular monomer . All the probed alpha-helices and most of the beta-structure was intact . There was an increase in the rate but not the extent of the mobilities of six of the probed tryptophan side chains, indicating loss of tertiary structure . Circular dichroism (CD) showed all the secondary structure to be intact . Intermediate III is monomeric and still globular, but the tryptophan anisotropy indicated an increase mobility at positions 36, 85, 190, 203, 279, and 285 . Helix alpha-B is further disrupted but helices alpha-1F, alpha-2G, and alpha 3G were still rigid . CD showed half the secondary structure to be still intact . Intermediate IV is a random coil in which all tryptophans have complete rotational freedom and the helix CD signal is lost.(ABSTRACT TRUNCATED AT 250 WORDS)

J Biol Chem, 1991 Jan 15, 266(2), 880 - 5
Cloning, sequencing, and overexpression of genes for ribosomal proteins from Bacillus stearothermophilus; Ramakrishnan V et al.; Although a low resolution model for the arrangement of the proteins of the small and large ribosomal subunits is known, a detailed mechanistic understanding of the function of the ribosome awaits a high resolution structure of its components . While crystals have been obtained of several ribosomal proteins from Bacillus stearothermophilus, determination of atomic resolution structures of these proteins is impeded by the difficulty of obtaining large amounts of native proteins for crystallographic or NMR studies . We describe here the cloning and overexpression in Escherichia coli of the genes for ribosomal proteins S5, L6, L9, and L18 from B . stearothermophilus . S5 is extremely toxic to E . coli when overexpressed, and we have taken advantage of a new tightly regulated expression system to obtain high yields (more than 100 mg of pure protein/liter of culture) of this protein . The B . stearothermophilus S5 produced in E . coli crystallizes, and the crystals are identical to those obtained from the native protein . The crystals diffract to 2-A resolution.

FEMS Microbiol Lett, 1991 Jan 15, 61(2-3), 271 - 5
Cloning of a chromosomal alpha-amylase gene from Bacillus stearothermophilus; Jorgensen PL et al.; We have cloned and sequenced a gene for a heat-stable alpha-amylase from a natural isolate of Bacillus stearothermophilus . Previously, it had been shown that B . stearothermophilus amylase genes may be harboured on indigenous plasmids . We have found that our isolate harbours the amylase gene only on the chromosome and not on its indigenous plasmid.

J Biol Chem, 1991 Jan 15, 266(2), 1170 - 6
Mechanism of inhibition of adenylate cyclase by phospholipase C-catalyzed hydrolysis of phosphatidylcholine . Involvement of a pertussis toxin-sensitive G protein and protein kinase C; Diaz-Laviada I et al.; The phospholipase C-mediated hydrolysis of phosphatidylcholine has been shown recently to be activated by a number of agonists . Muscarinic receptors, which trigger various signal transduction mechanisms including inhibition of adenylate cyclase through Gi, have been shown to be potent stimulants of this novel phospholipid degradative pathway . We demonstrate here, by exogenous addition of Bacillus cereus phosphatidylcholine-hydrolyzing phospholipase C, that phosphatidylcholine breakdown mimics the ability of carbachol to inhibit adenylate cyclase . This effect is sensitive to pertussis toxin and is entirely dependent on the presence of protein kinase C . This kinase is also required for the inhibition by carbachol of adenylate cyclase . These results suggest that the activation of phosphatidylcholine breakdown by phospholipase C may play an important role linking or favoring the coupling muscarinic receptors to Gi . Results presented here also show that phospholipase C-mediated hydrolysis of phosphoinositides by exogenous addition of Bacillus thuringiensis phosphoinositide-hydrolyzing phospholipase C does not affect adenylate cyclase, despite the fact that protein kinase C is translocated to an extent similar to that produced by the hydrolysis of phosphatidylcholine . According to the results shown here, both phospholipases also differ in their ability to down-regulate protein kinase C as well as to phosphorylate p80 and to transmodulate the binding of epidermal growth factor, two well established effects of protein kinase C in Swiss 3T3 fibroblasts . This emphasizes the complexity, from a functional point of view, of protein kinase C activation "in vivo."

Carbohydr Res, 1991 Jan 15, 209, 145 - 53
On the role of histidine residues in cyclodextrin glycosyltransferase: chemical modification with diethyl pyrocarbonate; Bender H; Ethoxyformylation with diethyl pyrocarbonate of approximately 1.5 His residues per molecule of enzyme reduced the cyclising activity of both the alpha-cyclodextrin glycosyltransferase from Klebsiella pneumoniae strain M 5 al and the beta-cyclodextrin glycosyltransferase from Bacillus circulans strain 8 by greater than 90% . Pre-incubation with substrate protected the enzymes from ethoxyformylation . Digestion of starch by the modified enzymes resulted in a delayed formation of cyclodextrins (cyclomalto-oligosaccharides, CDs), but a marked increase in the production of reducing saccharides . Similarly, coupling of alpha CD and maltose and successive disproportionation yielded mainly glucose and malto-oligosaccharides . The results are discussed in the context of the role of conserved His residues for binding of substrate and the transfer reactions.

Dev Comp Immunol, 1991 Winter, 15(1-2), 27 - 32
Antibacterial activity of Eisenia fetida andrei coelomic fluid: III--Relationship within the polymorphic hemolysins; Roch P et al.; The antibacterial activity exhibited by 10 different hemolytic, genetic families was established by measuring the inhibition of spontaneous in vitro growth by cell-free coelomic fluid toward 2 bacteria which are pathogenic for the earthworm: Bacillus megaterium (Gram +) and Aeromonas hydrophila (Gram -) . Only two families (B and K) displayed potent inhibitory activities . This finding is consistent with the fact that the B family occurs most frequently in both natural as well as in industrial breedings . Nevertheless, evidence of a poor antibacterial defense in some frequent families suggests the existence of alternative antibacterial mechanisms.

Salud Publica Mex, 1991 Jan-Feb, 33(1), 70 - 6
{Tuberculous meningitis: a 10-year analysis at the "Dr . Federico Gómez" Children's Hospital of Mexico}; Karam Bechara J et al.; Of all the forms of tuberculous infection in children, the most frequent is the pulmonary, but the tuberculous meningitis is the most severe and mortal, it occurs mainly in children under five years old, and the highest mortality is in children under two . The results of a retrospective study carried out at the Children Hospital of Mexico "Dr . Federico Gomez" about all the patients hospitalized with tuberculous meningitis diagnose during the January 1975 to December 1985 period were presented . One hundred and eighteen cases were studied: the majority (80%) corresponded to children under four years old, a percentage of 79 presented some degree of malnutrition, and 86 per cent showed clinical data of neurological affection . The confirmation of the diagnose was made in the majority of cases through laboratory and cabinet studies . Forty per cent showed no alteration in the chest X-rays, and the isolation of the bacillus was in a very low percentage (15%) . All the patients were treated with the antituberculous drugs mentioned, with a better development in the ones associated with steroids . The hospital stay was over 30 days in 15 per cent of the cases . The mortality of the series reviewed was 44.5 per cent.

J Gen Microbiol, 1991 Jan, 137 ( Pt 1), 41 - 8
Purification and properties of an acid endo-1,4-beta-glucanase from Bacillus sp . KSM-330; Ozaki K et al.; A novel acid cellulase (endo-1,4-beta-glucanase, EC 3.2.1.4) was found in a culture of Bacillus sp . KSM-330 isolated from soil . One-step chromatography on a column of CM-Bio-Gel A yielded a homogeneous enzyme, as determined by silver staining of both sodium dodecyl sulphate (SDS) and nondenaturing gels . The enzyme had a molecular mass of 42 kDa, as determined by SDS-polyacrylamide gel electrophoresis . The isoelectric point was higher than pH 10 . The N-terminal amino acid sequence of the enzyme was Val-Ala-Lys-Glu-Met-Lys-Pro-Phe-Pro-Gln-Gln-Val-Asn-Tyr-Ser-Gly-Ile-Leu- Lys-Pro . This enzyme had an optimum pH for activity of 5.2, being active over an extremely narrow range of pH values, from 4.2 to 6.9; below and above these pH values no activity was detectable . The optimum temperature at pH 5.2 was around 45 degrees C . The enzyme efficiently hydrolysed carboxymethylcellulose (CMC) and lichenan, but more crystalline forms of cellulose, curdlan, laminarin, 4-nitrophenyl-beta-D-glucopyranoside and 4-nitrophenyl-beta-D-cellobioside were barely hydrolysed . The enzymic activity was inhibited by Hg2+ but was not affected by other inhibitors of thiol enzymes, such as 4-chloromercuribenzoate . N-ethylmaleimide and monoiodoacetate . N-Bromosuccinimide abolished the enzymic activity, and CMC protected the enzyme from inactivation by this tryptophan-specific oxidant . It is suggested that a tryptophan residue(s) is involved in the mechanism of action of the Bacillus cellulase and that the inhibition of enzymic activity by Hg2+ is ascribable to interactions with the tryptophan residue(s) rather than with thiol group(s).

Pharmacotherapy, 1991, 11(2 ( Pt 2)), 64S - 71S
Monotherapy versus combination antimicrobial therapy: a review; Barriere SL; In view of the newer antibiotics that can cover the spectrum of organisms in mixed infections, single agents are now a viable option for antimicrobial therapy . In addition, monotherapy is relatively nontoxic and possibly less costly than combination therapy . Combinations may be more effective in preventing the emergence of resistance, however, and can also provide synergistic effects . They are a necessity in mycobacterial infections, enterococcal endocarditis, and deep-seated pseudomonal infections, as well as in patients with gram-negative bacillary infection with profound granulocytopenia.

Exp Pathol, 1991, 41(3), 147 - 50
Bacillus Calmette-Guérin (BCG) influences cell proliferation and glycosaminoglycans of chondrocyte cultures; Kittlick PD et al.; There are only few reports on the correlation between bacterial products and the GAG pattern of cartilage . Mycobacteria bovis (BCG) were applied to chondrocyte monolayer cultures for one week . The following parameters did change: cell proliferation increased, glycosaminoglycan synthesis and secretion decreased, hyaluronic acid in secreted and cell-associated glycosaminoglycans increased, a correlation between the degree of these changes and the degree of cell differentiation seems to exist . The contact of bacteria like BCG to chondrocytes may change the cellular metabolism . On the tissue level this may injure articular cartilage and thus support the concept of predamaged cartilage that is readily susceptible to further degradation.

Acta Otorrinolaringol Esp, 1991 Jan-Feb, 42(1), 75 - 7
{Primary laryngeal tuberculosis caused by Mycobacterium bovis}; Contreras Sanchez JD et al.; It is suggested that laryngeal tuberculosis is a common complication of pulmonary tuberculosis . The most frequent germ is Mycobacterium tuberculosis . We present a case in which a infrequent bacillus, Mycobacterium bovis, was isolated without lung afectation . Considerations about morphologic and microbiologic findings are discussed.

Urol Res, 1991, 19(1), 35 - 8
Induction of controlled prostatic tissue necrosis by bacille Calmette-Guérin derivatives; Morales A et al.; Intraprostatic administration of live bacille Calmette-Guerin (BCG) in humans has been found to produce tumor necrosis; unfortunately, the number of severity of complications have made its clinical use prohibitive . Previous studies have shown that soluble and microparticulate components present in the supernatants obtained after centrifugation of a reconstituted BCG preparation exhibit similar immunogenicity to the one shown by live bacteria . The supernatants, however, are not associated with disseminated infection of the progressive regional tissue destruction observed with the use of viable vaccine . Experiments were conducted to determine the effect of intraprostatic injection of BCG and its supernatants . Adult dogs, after positive conversion to protein purified derivative (PPD), were randomly assigned to three groups . Under direct vision and with digital rectal control, intraprostatic injections of various agents were given as follows: group I, normal saline; group II, live BCG; group III, 200 micrograms of BCG supernatants . Two months later the animals were sacrificed, and the prostates removed in toto and submitted for a thorough histological examination . Extensive but variable tissue necrosis was noted in groups II and III . No histological alterations were present in group I . The histological picture of the animals receiving BCG supernatants conclusively demonstrated circumscribed necrosis of the gland . Side effects and complications were present in animals receiving live BCG but conspicuously absent in the ones receiving supernatants . The observed effectiveness and safety of BCG supernatants for intraprostatic administration in an experimental system may lead to a simple, safe, and efficacious therapeutic modality for localized carcinoma of the prostate in humans.

Food Addit Contam, 1991 Jan-Feb, 8(1), 65 - 9
Residues of doxycycline and oxytetracycline in eggs after medication via drinking water to laying hens; Yoshimura H et al.; Doxycycline (DOTC) and oxytetracycline (OTC) were dissolved in drinking water (0.5 g/l) and supplied to laying hens for 7 consecutive days . Eggs laid were collected daily during and after medication, and the antibiotic concentrations in the yolk and albumin were determined by the cup-plate method with Bacillus cereus var . mycoides ATCC 11778 . The concentrations of both antibiotics were increased in yolk day by day with the advance in medication, reached peaks 2 days after withdrawal and then declined gradually . Mean peak concentrations in the yolk were 6.70 micrograms/g for DOTC and 1.42 micrograms/g for OTC . Peak concentrations in the albumin occurred in the middle stage of medication, where the mean values were 12.24 micrograms/g for DOTC and 1.03 micrograms/g for OTC . DOTC was detected in albumin until 24 days after withdrawal and for 2 days more in yolk than in albumin . OTC was detected in yolk until 9 days after withdrawal . The depletion period of OTC was shorter for the albumin, where the residue disappeared in all eggs 6 days after withdrawal . In spite of similarities between DOTC and OTC in structure, DOTC was deposited in higher concentrations and lasted for a longer period in eggs . This characteristic was considered due to its greater lipophilicity, closely correlated with oral absorption and tissue penetration.

J Invertebr Pathol, 1991 Jan, 57(1), 82 - 93
Peptide mapping of different Bacillus thuringiensis toxin gene products by CNBr cleavage in SDS-PAGE gels; Pang AS et al.; A cyanogen bromide fragmentation reaction of 3 hr in sodium dodecyl sulfate gel slices was used for the degradation of the toxins coded by the three cryIA genes from Bacillus thuringiensis kurstaki . Peptide patterns diagnostic for each toxin gene product were observed . Treatment longer than 3 hr led to the weakening and disappearance of protein bands . In 9 out of 16 wild B . thuringiensis strains tested, it was determined that only one of the cryIA genes was being expressed; in 6 strains, one to two genes were identified, with the presence of an additional gene possible and masked because of the overlap of peptide bands; in one strain none of the genes was expressed.

J Invertebr Pathol, 1991 Jan, 57(1), 101 - 8
Chemical modification of Bacillus thuringiensis subsp . thuringiensis (HD-524) trypsin-activated endotoxin: implication of tyrosine residues in lepidopteran cell lysis; Yan XJ et al.; A purified protein fraction from a solubilized and trypsin-digested extract of Bacillus thuringiensis subsp . thuringiensis (HD-524) fermentation powder was lytic to cells from several lepidopteran lines . Maximum yield was obtained by alkaline carbonate-thiocyanate solubilization of washed powder followed by trypsin digestion and Sephacryl (S-300) chromatography . The alkaline carbonate-solubilized fraction consisted predominantly of two bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with MW of 144 +/- 0.9 kDa and 134 +/- 1.4 kDa . After trypsin treatment and column chromatography, the cytolytic fraction consisted of a major band with a MW of 60.0 +/- 1.8 kDa and a minor band of 69 +/- 0.9 kDa . Cells from Trichoplusia ni (TN368) were most susceptible to lysis with 50% of cells lysed at 3 micrograms/ml, followed by Spodoptera frugiperda cells (SF21AE) exhibiting 50% cell lysis at 5 micrograms/ml and Lymantria dispar cells (Ld652Y) showing 40% lysis at 10 micrograms/ml . Chemical modification of the polypeptides was performed to determine the role of certain amino acid residues in the cytolytic activity . The group-specific reagent tetranitromethane was used to nitrate and oxidize tyrosine and cysteine residues, respectively . Cysteine residues alone were also modified with p-hydroxymercuribenzoic acid . Lysine residues were modified with O-methylisourea . Of the three types of amino acid residues, only the modification of tyrosine resulted in reduced cell lysis.

Cancer Immunol Immunother, 1991, 32(5), 315 - 24
Hypotension and disseminated intravascular coagulation following intralesional bacillus Calmette-Guérin therapy for locally metastatic melanoma; Cohen MH et al.; Four patients developed serious hypotension and signs of disseminated intravascular coagulation shortly after a second round of Tice bacillus Calmette-Guerin (BCG) injections into locally recurrent cutaneous melanoma satellite nodules . Each of these patients survived following intensive therapy with isoniazid, pyridoxine, steroids, pressors, antibiotics, and cardio-renal support including, in one case, three acute hemodialyses . Plasma specimens from two of the four patients caused gelation of lysate from the amebocytes of Limulus polyphemus, indicating the presence of endotoxin or an endotoxin-like substance . In vitro studies on the BCG preparations led us to conclude that this endotoxin activity in the plasma is not the result of direct injection of endotoxin with the BCG preparation, but rather from release of endotoxin from endogenous sources, such as the intestinal tract during a period of relative hypotension following an allergic reaction . Prior immunity appeared to be the consistent factor in the toxic reactions reported herein . Finally, we present recommendations for serial monitoring of these patients and discuss the use of an alternative agent for intralesional therapy.

Chest, 1991 Jan, 99(1), 123 - 7
The chest roentgenogram in pulmonary tuberculosis patients seropositive for human immunodeficiency virus type 1; Long R et al.; To determine the impact that co-infection with HIV has on the radiographic presentation of pulmonary tuberculosis, we examined the chest roentgenograms obtained before treatment in 225 HIV-tested adult Haitians with bacillary (smear or culture or both) positive pulmonary tuberculosis . There were 67 HIV-seropositive and 158 HIV-seronegative patients . Intrathoracic adenopathy alone was more common and parenchymal infiltrates less common in HIV-seropositive patients (p less than 0.05) . Although a parenchymal infiltrate was less likely to be cavitating in the HIV-seropositive group (p less than 0.05) when cavitary parenchymal disease was present, HIV seropositivity did not affect the number of cavities (single or multiple) or the size of the largest cavity . Patients with AIDS were significantly more likely to have a chest radiographic pattern consistent with primary tuberculosis (80 percent) than HIV-seropositive patients without AIDS (30 percent), and the latter were significantly more likely to have such a pattern than HIV-seronegative patients (11 percent) (p less than 0.05) . The HIV-seropositive patients were equally infectious, regardless of the pattern of disease (primary vs postprimary) . Even though pulmonary tuberculosis in an HIV-seropositive adult probably results from reactivation of dormant foci or reinfection, the pattern on the chest roentgenogram often suggests primary disease, especially if the patient has AIDS.

Anesth Analg, 1991 Jan, 72(1), 94 - 100
Clinical evaluation of autotransfusion during liver transplantation; Kang Y et al.; The clinical suitability of intraoperative autotransfusion was evaluated in 25 patients undergoing orthotopic liver transplantation using a Cell Saver #4 (Haemonetics) with acid-citrate-dextrose anticoagulation . In the first 14 patients (phase 1), biochemical, hematologic, coagulation, and semiquantitative bacteriologic studies were performed from the collected blood, processed blood, and patients' blood before and after 500 mL of autotransfusion . The acid-citrate-dextrose solution produced adequate anticoagulation, and the system effectively removed most of the potassium, red blood cell fragments, plasma free hemoglobin, bilirubin, coagulation factors, platelets, and fibrin degradation products . Autotransfusion (500 mL) did not alter coagulation, electrolyte balance, and hematologic findings in recipients except for a clinically insignificant increase in plasma free hemoglobin . Seventeen of 56 samples of the collected blood or processed blood were positive for coagulase (-) Staphylococcus (occasional or rare), but blood cultures before and after autotransfusion were negative in all patients . In the next 11 patients (phase 2), a quantitative bacteriologic study was performed from the collected blood, processed blood, skin, bile duct stump, peritoneal cavity, and room air using a mock reservoir . The processed blood was not transfused . All blood cultures from the patients were sterile . However, coagulase (-) Staphylococcus or Bacillus sp was seen in two cultures from skin, three from the processed blood, and three from air, suggesting that room air and skin were the sources of contamination . When the patients of the two phases of study were compared, postoperative blood cultures were all sterile, and renal function was similar . Therefore, autotransfusion appears to be clinically acceptable during liver transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

J Urol, 1991 Jan, 145(1), 40 - 3; discussion 43-4
Progression of stage T1 bladder tumors after intravesical bacillus Calmette-Guerin; Herr HW; Of 221 patients with superficial bladder tumors (stages Ta, Tis and T1) treated by transurethral resection and an initial 6-week course of intravesical bacillus Calmette-Guerin (BCG) 195 were evaluated after 3 months for local response . Of these patients 17 (8.7%) had a stage T1 tumor, of whom 14 (82%) subsequently had muscle invasion (7), uncontrolled local disease (6) or metastasis (1) . The median interval to progression was 8.4 months, which was significantly (p = 0.0001) less than among the other 178 patients with a nonstage T1 lesion or no tumor found at 3 months, of whom 44 (25%) had progression (median more than 5 years) . These data suggest that patients with stage T1 tumor 3 months after BCG therapy require additional therapy other than simple tumor resection and meticulous followup.

J Urol, 1991 Jan, 145(1), 195 - 8
Immunotherapy of murine bladder cancer by irradiated tumor vaccine; Lamm DL et al.; This investigation explored the efficacy of irradiated autologous mouse bladder tumor (Ir-MBT2) as an active specific immunotherapeutic agent and as adjuvant therapy with Bacillus Calmette-Guerin (BCG) against a subcutaneously transplanted murine bladder tumor . Tumor incidence was significantly reduced in groups receiving BCG (27%, p less than 0.005) or Ir-MBT2 with BCG (53%, p less than 0.025), compared to control (93%) . Survival was significantly improved in groups treated with BCG (100%, p less than 0.005), 10(5) Ir-MBT2 with BCG (53%, p less than 0.01), or 10(7) Ir-MBT2 with BCG (47%, p less than 0.025) compared with control (13%) . Surprisingly, Ir-MBT2 consistently reduced the efficacy of BCG alone . Ir-MBT2 alone (10(7)) appeared to enhance tumor growth . Autologous irradiated bladder tumor vaccine, alone or in combination with BCG, displayed no immunotherapeutic advantage . The use of irradiated tumor cell vaccine for bladder cancer therapy may reduce the results achievable with BCG alone.

Arch Microbiol, 1991, 156(3), 181 - 5
Influence of an S-layer on surface properties of Bacillus stearothermophilus; Gruber K et al.; Various aspects of surface properties of the S-layer-carrying Bacillus stearothermophilus PV72 and of an S-layer-deficient mutant (strain PV72/T5) have been tested by adsorption assays on solid surfaces, electrostatic interaction chromatography and hydrophobic interaction chromatography . The adsorption assays have shown that cell adhesion of the S-layer-carrying strain was less influenced by environmental changes than it was with the S-layer-deficient mutant . Electrostatic interaction chromatography indicated that both strains have positively and negatively charged groups exposed on the cell surface but the S-layer-carrying strain reveals more positively charged groups than does the S-layer-deficient mutant . Hydrophobic interaction chromatography showed that both strains have a hydrophilic surface but that the hydrophilic properties are more pronounced with the strain lacking an S-layer.

Arch Microbiol, 1991, 155(6), 526 - 34
Molecular cloning, structure, promoters and regulatory elements for transcription of the Bacillus licheniformis encoded regulon for xylose utilization; Scheler A et al.; In this article we describe the cloning of the xyl regulon encoding xylose utilization from Bacillus licheniformis by complementation of a xyl mutant of B . subtilis . The xylose isomerase encoding gene, xylA, was sequenced and identified by its extensive homology to other xylose isomerases . The expression of xylA is regulated on the level of transcription by a repressor protein encoded by xylR . Its gene has the opposite orientation of xylA and the start codons are 181 bp apart . A deletion of xylR renders xylA expression constitutive . The xylR sequence was determined and is discussed with respect to its homology to other xylR structures . Primer extension analyses of the xylA and xylR transcripts under repressing and including conditions define their promoters and confirm the regulation of xylA transcription . Furthermore, some induction of the xylR transcript by xylose is also observed . The regulatory sequence of both genes consists of a bipolar promoter system and contains three palindromic sequence elements . Their potential functions with respect to xylA and xylR regulation are discussed . The primary structures of the genes, promoters and regulatory sequences are compared to the xyl regulons encoded by B . subtilis, B . megaterium, Staphylococcus xylosus and E . coli . Homology is greatest between the B . subtilis and B . megaterium encoded xyl genes while the B . licheniformis borne genes are clearly more distant . The next greater differences are found to the S . xylosus and the greatest to the E . coli encoded genes . These results are discussed with respect to the taxonomic relations of these bacteria.

Urol Int, 1991, 47 Suppl 1, 80 - 2
Development of bacillus Calmette-Guérin-induced antitumor activity in peripheral blood mononuclear cells; Koga S et al.; Peripheral blood mononuclear cells (PBMNCs) after bladder instillation of bacillus Calmette-Guerin (BCG) to patients with superficial bladder carcinoma showed lytic activity against natural killer (NK)-sensitive and NK-resistant target tumor cells . PBMNCs in normal volunteers were isolated and cultured with BCG (5 micrograms/ml) in vitro . Maximum cytotoxicity was obtained within 24 h . Cytosine arabinoside as a DNA synthesis inhibitor did not inhibit this killer cell activation . Monoclonal antibody treatment revealed that both precursor and effector cells were NK cell type . However, effector cells had a wider spectrum of target cells than usual NK cells . The recognition specificity from cold target competition experiments was selective.

Urol Int, 1991, 47 Suppl 1, 104 - 7
Combined therapy of interleukin 2 with cyclophosphamide or bacillus Calmette-Guérin against implanted bladder cancer cells in mice; Wada S et al.; 1 x 10(5) cells of FAMFT-induced bladder cancer cell line (MBT-2) were injected into the right leg of the C3H/He mice . 10 days later, these animals with the same tumor size were divided into four groups . Group 1 is the control with no treatment . Group 2 underwent interleukin 2 (IL-2) treatment for 10 days since the 11th day after tumor inoculation . Group 3 received just one CPM injection on the 10th day . Group 4 had a combination therapy of CPM, followed by IL-2 . Group 1 and 2 exhibited enlargement of tumor size, resulting in the death of all animals . Administration of CPM led to temporary disappearance of tumor, followed by local tumor recurrence in all cases . However, combined therapy with IL-2 showed complete remission of the tumors . Natural killer cell activity measured by the 51Cr release assay of the spleen cells using Yac-1 cells as target cells 20 days after tumor inoculation was found to be significantly higher in group 2 and 4, while lymphokine-activated killer cell activity of the spleen cells against P-815 cells showed no significant difference between the four groups.

In Vivo, 1991 Jan-Feb, 5(1), 71 - 8
Immunosuppression by an immunogenic immunomodulating mycobacterial fraction is correlated to changes in phenotype distribution of murine lymphoid cells; Rashid G et al.; Previously reported studies revealed that extensive immunization with the Methanol Extraction Residue (MER) of BCG tubercle bacillus in Incomplete Freund's Adjuvant (IFA) induced marked suppression of T and B cell functions in vivo and in vitro . The purpose of the present work was to determine whether immunosuppression induced by hyperimmunization with MER is correlated with changes in morphological characteristics and in phenotype pattern of spleen and peritoneal lymphoid cells . Hyperimmunization with MER resulted in a marked increase in the number of spleen cells and in enlargement and granulation of spleen and peritoneal cells . Similar changes in size and granulation were also observed in isolated fractions of splenic T and B cells . Extensive immunization with MER also induced marked decrease in the total number of T cells (Thy 1,2 positive) . The decrease in T cells was observed in all three T-cell subsets investigated: Lyt-1, Lyt-2 and L3T4 positive . Although the number of splenic B cells was decreased in samples (10,000 cells), taken from MER hyperimmunized mice, this decrease was compensated by overall increase in the number of spleen cells . The marked decrease in the percentage of splenic T cells was counterbalanced by marked increase in the splenic macrophage population: increase in MAC-1, MAC-2 and MAC-3 positive cells . It is concluded that extensive immunization with MER induces morphological changes in spleen and peritoneal cells, marked decrease in the number of splenic T cells and marked increase of the splenic macrophagic population . It is postulated that these changes are correlated with induction of immunosuppression by a similar procedure of extensive immunization with the agent.

Appl Biochem Biotechnol, 1991 Spring, 28-29, 699 - 718
A structured model for vegetative growth and sporulation in Bacillus thuringiensis; Starzak M et al.; A mathematical model has been developed for the delta-endotoxin producing Bacillus thuringiensis . The structure of the model involves the processes taking place during vegetative growth, those leading to the initiation of sporulation under conditions of carbon and/or nitrogen limitation, and the sporulation events . The key features in the model are the pools of compounds, such as PRPP, IMP, ADP/ATP, GDP/GTP, pyrimidine nucleotides, NAD/NADH2, amino acids, nucleic acids, cell wall, and vegetative and sporulation proteins . These, along with sigma-factors that control the nature of RNA-polymerase during the different phases, effectively stimulate the vegetative growth and sporulation . The initiation of sporulation is controlled by the intracellular concentration of GTP . Results of simulation of vegetative growth, initiation of sporulation, spore protein formation, and production of delta-endotoxin under C- or N-limitation are presented.

Probl Tuberk, 1991, (5), 23 - 5
{Helium-neon laser in the complex treatment of patients with pulmonary tuberculosis}; Shesterina MV et al.; The paper gives the results of helium-neon laser irradiation of the endobronchus used in the combined treatment of 30 patients with newly-diagnosed pulmonary tuberculosis complicated by nonspecific endobronchitis . Inclusion of endobronchial phototherapy and helium-neon laser radiation in the complex treatment of patients with newly diagnosed pulmonary tuberculosis complicated by nonspecific endobronchitis makes it possible to achieve clinical cure of an inflammatory bronchial process by 35% more frequently and over 2-3 times more rapidly than in its absence and to increase treatment efficiency of the main process; the terms of bacillary excretion decrease by 1 month, on the average, closure of destruction cavities (with an increase in the parameter by 17%) by 1.2 months and hospital treatment by 1.2 months, as compared with the respective parameters obtained by the traditional treatment of the similar patients.

Exp Pathol, 1991, 42(3), 145 - 50
Effect of the microbial constituents, LPS and BCG, on the glycosaminoglycans of chondrocyte cultures; Kittlick PD et al.; Earlier we reported that articular chondrocytes in monolayer culture produce pericellular proteoglycans both with short and long half lives, T-1 and T-2 (Kittlick et al . 1991 b) . Now monolayer cultures have been investigated to assess the influence on the metabolism of pericellular proteoglycans or glycosaminoglycans by lipopolysaccharide of E . coli and S . typhimurium as well as by Bacillus Calmette-Guerin.

Eur Urol, 1991, 19(4), 319 - 21
Effect of distilled water and bacillus Calmette-Guérin on exfoliated bladder tumor cells; Bolkier M et al.; The effect of distilled water on exfoliated tumor cells from transitional bladder carcinoma was compared to the effect of bacillus Calmette-Guerin (BCG; a nonspecific immunotherapeutic agent) dissolved in either saline or distilled water . The experiment was controlled by comparison to the effect of saline on tumor cells under the same conditions . Distilled water proved as effective as BCG in distilled water (p less than 0.001) and much more destructive than BCG in saline within the 2-hour time limit of the experiment (p less than 0.001).

Pathobiology, 1991, 59(3), 148 - 52
New insights into the immunopathology of tuberculosis; Rook GA et al.; Tuberculosis is characterised by fever, weight loss and necrosis in both lesions and tuberculin skin test sites (Koch phenomenon), although the antigens of Mycobacterium tuberculosis are not directly toxic to the tissues . The tissue damage appears to be due to several interacting factors . First, M . tuberculosis induces an immunoregulatory disorder of which a raised percentage of agalactosyl IgG is a marker . This is seen also in rheumatoid arthritis and Crohn's disease and is associated with tissue-damaging inflammation . Subsequently, several properties of M . tuberculosis exacerbate this disorder by triggering cytokine release, and rendering tissues sensitive to the toxicity of tumor necrosis factor (TNF) . Moreover, M . tuberculosis, but not bacillus Calmette-Guerin or several Mycobacterium avium strains, produces a factor which increases the toxicity of TNF for individual cells . Thus, M . tuberculosis may distort the normal protective role of TNF so that this cytokine becomes toxic to the host . The immunoregulatory disorder associated with agalactosyl IgG appears to be susceptible to immunotherapy, so novel types of treatment for the immunopathological component of tuberculosis are being explored.

Scand J Infect Dis, 1991, 23(1), 71 - 7
Bacillus cereus: a significant pathogen in postoperative and post-traumatic wounds on orthopaedic wards; Akesson A et al.; Over a period of 19 months, Bacillus species, most frequently B . cereus, were isolated from postoperative and post-traumatic wounds in 24 patients on orthopaedic wards in a Swedish hospital . Clinical signs of infection were found in all but 3 patients: in 9 patients, mild signs with increased secretion from the wounds; in 10 patients, moderate signs with fever and/or significant local reaction around the wounds; and in 2 patients, severe signs with necrotic infections . Bacillus spp . were isolated from one-fourth of all patients with wound complications following total hip arthroplasty . Their hospitalization was significantly prolonged, compared with a control group (p = 0.0042) . Isolates of B . cereus from wounds should not be dismissed as contaminants or as non-pathogenic, but must be assessed in the light of the clinical situation.

Antimicrob Agents Chemother, 1991 Jan, 35(1), 201 - 2
In vitro susceptibilities of ocular Bacillus cereus isolates to clindamycin, gentamicin, and vancomycin alone or in combination; Gigantelli JW et al.; A broth dilution assay was used to determine the in vitro susceptibilities of 10 ocular isolates of Bacillus cereus to clindamycin, gentamicin, and vancomycin both alone and in combination . The checkerboard technique was used to determine fractional inhibitory and bactericidal concentration indices for combinations of clindamycin-gentamicin and vancomycin-gentamicin.

Infect Immun, 1991 Jan, 59(1), 65 - 72
Molecular cloning, sequencing, and identification of a metalloprotease gene from Listeria monocytogenes that is species specific and physically linked to the listeriolysin gene; Domann E et al.; The entire nucleotide sequence of an open reading frame located immediately downstream of the listeriolysin gene from a virulent Listeria monocytogenes serotype 1/2a strain was determined . The product of the open reading frame was 510 amino acids with a predicted molecular weight of 57,400 . The deduced amino acid sequence of this open reading frame is highly similar to that of a family of secreted metalloproteases produced by various members of the genus Bacillus, of which thermolysin is the prototype . Immunoblots performed with specific antisera raised against thermolysin from Bacillus stearothermophilus allowed the detection of a 60-kDa polypeptide, corresponding to the pro-form of the protease, in culture supernatants of L . monocytogenes strains . In maxicell experiments, Escherichia coli recombinants harboring this open reading frame also specifically directed production of a 60-kDa protein . Protease activity was low to undetectable in both Listeria strains and E . coli recombinants . This is due to lack of processing of the inactive pro-form of the protease to its mature active form in both species . We have designated this gene mpl for metalloprotease of L . monocytogenes . The gene was present only in pathogenic L . monocytogenes strains, in which it was physically linked to the listeriolysin gene.

Am Rev Respir Dis, 1991 Jan, 143(1), 69 - 73
Impact of human immunodeficiency virus type 1 on tuberculosis in rural Haiti; Long R et al.; We conducted a case-control study to determine the relative and attributable risk of HIV seropositivity for bacillary-positive (smear and/or culture) pulmonary tuberculosis in Haiti . There were 274 patients with tuberculosis and an equal number of control subjects . Antibodies to HIV were present in 67 (24%) patients and eight (3%) control subjects . Odds ratios suggested that the risk of pulmonary tuberculosis was 15.7 times as great (95% confidence interval, 4.8 to 5.0; p less than 0.05) in patients 20 to 39 yr of age who were HIV-seropositive than in HIV-seronegative patients . In contrast, the relative risk in those 40 to 59 yr of age was elevated (3.0 times), though not significantly (lower 95% confidence interval, 0.8) . In the 20- to 39-yr age group, 31% of tuberculosis was attributable to HIV infection (95% confidence interval between 23 and 39%) . HIV-seropositive and HIV-seronegative patients did not differ with respect to sputum smear positivity . HIV-seronegative patients were twice as likely to be infected with resistant organisms, though this was not significant . We conclude that HIV infection is a major risk factor for pulmonary tuberculosis in young adult residents of Haiti . This, together with the fact that similar proportions of HIV-seropositive and HIV-seronegative patients were potentially infectious, suggests that without vigorous counteraction tuberculosis will become a greater problem for Haiti.

J Gastroenterol Hepatol, 1991 Jan-Feb, 6(1), 67 - 70
Fatal amoebic liver abscess: an autopsy study; Ahmad M et al.; An autopsy study of 20 cases is presented . The mean age was 32 years (range: 14-45) . Four cases (20%) presented with jaundice . Another four (20%) had continuous fever with abdominal pain . Six had loose motions with blood and mucus in the stools . The mean duration of illness was 7.2 days . All cases were thin and emaciated . Liver was grossly enlarged (mean weight: 2680 g) . The abscess was single in all cases except one . The right lobe was involved in 15 cases, the left in four and both in one . The average size of abscess was 13 cm . The abscess had ruptured in the abdomen in 3 patients, for which laparotomy was performed . Pulmonary involvement was seen in 3 cases . Colon showed ulceration in half the patients . In 2 cases perforation was also present . No cerebral involvement was present . A number of interesting features emerged from this study . Although amoebic liver abscess is a common disease, its diagnosis can be missed due to unusual presentation . In the present study, diagnosis of viral hepatitis, carcinoma lung, bacillary dysentery and enteric fever with perforation were made, which probably contributed materially to fatal outcome.

Probl Tuberk, 1991, (3), 34 - 6
{Therapeutic possibilities of inhalation of rifampicin with dimexide in phthisiopulmonology}; Gorbach IN et al.; The efficacy of treatment with rifampicin-dimexide++ aerosols of 184 pulmonary tuberculosis patients and 18 chronic obstructive bronchitis patients was studied . The character of the residual changes following the main chemotherapeutic course was studied in 98 patients . Dimexide++ was proved to be a good rifampicin solvent and synergist enhancing its bacteriostatic action . Rifampicin used in a dose of 150-300 mg in 3-4 ml of dimexide++ in aerosols for 1-2 months promotes bacillary excretion cessation and healing of destruction cavities and allows for a reduction of its toxic action on liver parenchyma . Rifampicin--dimexide++ aerosols contribute to more complete tuberculosis cure and reduce the residual pulmonary and bronchial changes . Rifampicin with dimexide++ can be successfully used for treating chronic bronchitis both as an independent disease and concurrent disease in pulmonary tuberculosis.

Probl Tuberk, 1991, (3), 25 - 7
{Clinical course of infiltrative tuberculosis in adolescents}; Vladimirova MI et al.; The course of infiltrative tuberculosis making up 53.2% in the pattern of clinical forms was investigated in 64 adolescents . Infiltrative tuberculosis morbidity among the girls is 3 times higher . The main detection method is represented by a planned roentgenofluorographic examination, since in the region in question infiltrative tuberculosis tends to have limited symptoms in 56.2% of the adolescents, including 30.5% of them even with disseminated process . In 60.9% of the cases, infiltrative tuberculosis is observed to have a decomposition phase which is 4.5 times more frequently traced in patients living in contact with a bacillary ++carrier . In 8% of the patients, infiltrative tuberculosis was complicated by hemoptysis . Infiltrative tuberculosis in 32.8% of the adolescents manifests itself in a pulmonary form of primary tuberculosis . Due to a complex treatment hemodynamic indices are normalized by the end of the first month in addition to the removal of the intoxication signs by the third month . Cavity closure was achieved in 71.8% of the patients . Despite the given therapy, transformation into the destructive forms of tuberculosis took place.

Scand J Gastroenterol Suppl, 1991, 181, 58 - 64
Characterization of the morphologic conversion of Helicobacter pylori from bacillary to coccoid forms; Catrenich CE et al.; Growth studies of Helicobacter pylori were performed involving analysis of the bacterium and its microenvironment, to lend insight into the factors responsible for the morphologic conversion phenomenon . H . pylori converted from bacillary to coccoid forms in broth culture after incubation for 5 days under microaerobic conditions with agitation . This morphologic conversion was paralleled by a dramatic decrease in colony-forming units per milliliter (CFU/ml) and a significant endogenous increase in the pH of the broth culture . In addition, removal of broth cultures from microaerobic conditions after 3 days of incubation resulted in a rapid increase in culture pH, a morphologic conversion, and a concomitant decrease of CFU/ml . These observations suggest an inhibitory effect of basic pH, endogenously produced, on the growth of H . pylori in vitro . Experiments designed to identify the reason for the endogenous increase in culture pH demonstrated that the urease enzyme of H . pylori is not primarily responsible for this phenomenon . Rather, H . pylori appears to produce a deaminase enzyme that is likely responsible for the generation of ammonia, which results in the increase in culture pH, the morphologic conversion, and the loss of culturability observed in vitro . Also indicated is the need for a buffering component (for example, bicarbonate) to maintain pH conditions favorable to the growth of H . pylori.

Probl Tuberk, 1991, (4), 32 - 4
{Disseminated tuberculosis of the lungs in children}; Mitinskaia LA et al.; The infiltration phase of disseminated pulmonary tuberculosis has recently become a severe form of tuberculosis in children . It is accompanied by bacillary excretion (61.3%), lung tissue destruction (29.1%) and process generalization with extrapulmonary localization . Its course is most severe in the phase of destruction and has fatal outcome in 26.4% of cases which is mainly encountered in children of the early and pubertal age . The differential diagnosis of disseminated pulmonary tuberculosis in children with dissemination of non-tuberculous etiology is of importance considering a high percentage of diagnostic errors . Timely diagnosis of the destruction phase of disseminated pulmonary tuberculosis requires tuberculin diagnosis with Mantoux test with 2 TU PPD-L and bacteriological examination of all children with diffuse lung lesions staying in children's somatic hospitals.

Probl Tuberk, 1991, (4), 15 - 8
{New approaches to the treatment of mycobacterioses of the lungs}; Aleksandrova AE et al.; The paper deals with improvement of pulmonary mycobacterioses treatment . In the preliminary experiments with the use of an experimental model, 32 regimens of M . avium mycobacteriosis treatment were tested on 872 white mice and the most efficient one was chosen, which comprised a combination of kanamycin-ethambutol-thionamide coupled with sulfadimethoxine and thymalin courses . The clinical course and results of treatment of 43 patients (23 of the main and 20 of the control group) with fibro-cavernous pulmonary mycobacteriosis were studied . Employment of the new treatment regimen significantly enhanced its efficiency--the incidence of bacillary excretion cessation increased from 30 to 82.6%.

Mol Microbiol, 1991 Jan, 5(1), 197 - 205
Purification and characterization of two-subunit cytochrome aa3 from Bacillus cereus; Garcia-Horsman JA et al.; Cytochrome c-oxidase type aa3 (EC 1.9.3.1) was purified to homogeneity from vegetative Bacillus cereus by ion-exchange and hydroxylapatite chromatography in the presence of Triton X-100 . Gel filtration analysis suggested a dimeric structure apparently 172 kDa in size; however, only a monomer of 81 kDa was detected when analysed by non-denaturing gel electrophoresis . Denaturing gel electrophoresis analysis of the protein showed the presence of two subunits (51 and 30 kDa) . Atomic absorption and visible spectroscopy showed typical aa3 redox centres with haem a iron and copper in a ratio of 22 nmol and 35 ng-atom per mg protein, respectively . No haem c was found associated with the purified enzyme in the conditions reported here . Oxidase activity was fully reconstituted by phospholipids in the presence of N,N,N',N'-tetramethyl-p-phenylenediamine or reduced yeast cytochrome c (but not horse cytochrome c) as electron donors . This activity was abolished by cyanide and carbon monoxide.

Lab Anim Sci, 1991 Jan, 41(1), 26 - 30
Detection of serum antibodies to Bacillus piliformis in mice and rats using an enzyme-linked immunosorbent assay; Motzel SL et al.; Rats and mice were infected with Bacillus piliformis organisms at a dosage which resulted in clinical signs of Tyzzer's disease in gerbils . Although rats and mice did not show clinical signs of disease, rising antibody titers to B . piliformis were detected by enzyme-linked immunosorbent assay (ELISA) 2 to 6 weeks post-inoculation and remained at positive levels 11 weeks post-inoculation . Western blot analyses of sera from experimentally infected animals revealed banding patterns nearly identical to those obtained using hyperimmune serum . Results indicated that elevated ELISA titers reflected production of specific antibodies directed against antigens of B . piliformis . ELISA and Western blot analyses of naturally infected animals yielded similar results . These findings suggest that immunoassays such as ELISA can be used to detect subclinically infected rats and mice in the absence of clinical signs or histopathologic evidence of Tyzzer's disease.

Biochem J, 1991 Jan 1, 273(Pt 1), 43 - 7
The mechanism of sunlight-mediated inactivation of Bacillus thuringiensis crystals; Pusztai M et al.; Detailed photostability studies were carried out using purified delta-endotoxin crystals from Bacillus thuringiensis subspecies HD-1 and HD-73 . The mechanism and time course of sunlight inactivation was investigated by: (a) monitoring the tryptophan damage in the intact crystals by Raman spectroscopy, (b) amino acid analysis and (c) biological assays using insects . The results demonstrate that, for purified HD-1 or HD-73 crystals, the 300-380 nm range of the solar spectrum is largely responsible for bringing about crystal damage and consequent loss of toxicity . Purified Bacillus thuringiensis crystals that were exposed to fermentation liquor after cell lysis were more quickly degraded by sunlight than were crystals from cells that were lysed in water . This effect is attributed to adsorption of chromophores by crystals exposed to the fermenter liquor and the subsequent ability of these chromophores to act as photosensitizers . The importance of a photosensitization mechanism in crystal degradation was further emphasized by irradiating Bacillus thuringiensis crystals in vacuo . The latter crystals were found to be less damaged (20% tryptophan loss after 24 h irradiation by the solar spectrum) compared with crystals from the same sample irradiated in air (60% (60% tryptophan loss) . Other methods of decreasing exposure of the crystals to oxygen, e.g . by using glycerol as a humectant, were also found to be successful in controlling photodamage . The results concerning photodegradation support a photosensitization mechanism involving the presence of exogenous (and possibly endogenous) chromophores which create singlet oxygen species upon irradiation by light.

Am J Dis Child, 1991 Jan, 145(1), 98 - 101
Cat-scratch disease . Acute encephalopathy and other neurologic manifestations; Carithers HA et al.; Seventy-six patients with neurologic complications of cat-scratch disease are discussed . Encephalopathy occurred in 61, while 15 had either cranial or peripheral nerve involvement . The average age of the patients with encephalopathy was 10.6 years (range, 1 to 66 years), and almost twice as many males as females were affected in contrast to patients with uncomplicated cat-scratch disease, in which the ratio was almost equal . Fever was not documented in 50% of patients with encephalopathy and only 26% had temperatures higher than 39 degrees C . Convulsions occurred in 46% and combative behavior in 40% . Lethargy with or without coma was accompanied by variable neurologic signs . Results of laboratory studies, including imaging of the central nervous system, were inconsistent and nondiagnostic . Biopsy tissue from 14 patients showed histopathologic findings compatible with cat-scratch disease . The "English-Wear bacillus" was demonstrated by the Warthin-Starry stain in 10 of 14 skin or lymph node specimens . Of the 15 patients without encephalopathy, two children with facial nerve paresis displayed cranial nerve symptoms and/or signs, 10 patients had cat-scratch disease neuroretinitis, and three women had peripheral neuritis . All 76 patients recovered within 12 months; 78% recovered within 1 to 12 weeks . There were no neurologic sequelae . Treatment consisted of control of convulsions and supportive measures . Commonly used antibiotics administered to more than half of the patients were apparently ineffective.

Biotechnol Ther, 1991, 2(1-2), 159 - 78
Development of BCG as a live recombinant vector system: potential use as an HIV vaccine; Fuerst TR et al.; Bacille Calmette-Guerin (BCG), a live attenuated tubercle bacillus, is currently the most widely used vaccine in the world . Because of its unique characteristics, including low toxicity, adjuvant potential, and long-lasting immunity, BCG represents a novel vaccine vehicle with which to deliver protective antigens of multiple pathogens . We have developed episomal and integrative expression vectors employing regulatory sequences of major BCG heat shock proteins for stable maintenance and expression of foreign antigens in BCG vaccine strains (22) . Shuttle plasmids capable of autonomous replication in Escherichia coli and BCG were constructed with a DNA cassette containing a minimal replicon derived from the Mycobacterium fortuitum plasmid pAL5000 . Efficient and stable chromosomal integration of recombinant plasmids into BCG was achieved using a DNA segment containing the mycobacteriophage L5 attachment site and integrase coding sequence . Using the BCG hsp60 and hsp70 stress gene promoters, we were able to express Escherchia coli beta-galactosidase to levels in excess of 10% of total cell protein . The major antigens of HIV-1 gag, pol, and env were also stably expressed using our vector systems . The recombinant BCG elicited long-lasting humoral and cellular immune responses to these antigens in mice . Antibody responses to beta-galactosidase using as few as 200 colony-forming units were detected 6 weeks after immunization, and titers (1:30,000) were sustained for more than 10 weeks . Cellular immune responses, of both cytotoxic T cell (CTL) and helper T lymphocytes, were detected to beta-galactosidase . CTL responses were also induced to the HIV-1 envelope protein . Thus, we have demonstrated stable recombinant antigen expression, processing, and presentation using our recombinant BCG vector system . This live recombinant vector system shows promise as a universally applicable and safe vaccine vehicle for protection against various infectious diseases.

Wiad Parazytol, 1991, 37(4), 461 - 70
{Implications of biological pest and parasite control for environment}; Lipa JJ; Biological control of parasites, vectors and pests must consider safety to non-target organisms and to the environment . The WHO safety procedure was discussed in respect to such agents as Bacillus thuringiensis, Romanomermis culicivorax, Gambusia affinis and others . Monitoring of the environment in which biological control treatments are performed is necessary to secure safety requirements as well as the efficiency of biological control programmes.

Pneumonol Alergol Pol, 1991, 59(9-10), 48 - 54
{Tuberculosis in Poland and in the world--actual state and expectations after the year 2000}; Leowski J et al.; Decreased trends of tuberculosis incidence have been observed in Poland . A rate of decreased incidence has been hesitated from about 1% to above 12% per year . In 1989 year 16,185 (42.8%/100,000) cases with active pulmonary tuberculosis have been registered and about half of them have been bacteriologically supported . Analyzed epidemiological parameters for tuberculosis are twice higher in Poland than in Europe . Using relation between an incidence of active tuberculosis bacteriologically supported and annual risk of infection it was documented that an annual risk of tubercule bacillus infection was 10-times higher in Poland in comparison with other countries characterized by the lowest incidence of tuberculosis (Norway, Holland).

Folia Microbiol (Praha), 1991, 36(6), 590 - 1
Adsorption on bacterial spores depending on the aggregation properties of antimicrobial tensides; Cupkova V et al.; A change in interaction with spores of Bacillus cereus occurred in the range of critical concentrations of micelle formation . With 1-methyldodecyldimethylamine-N-oxide and N,N'-bis(dodecyldimethyl)-1,2-ethanediammonium dibromide, the induced release of dipicolinic acid was blocked and the adsorption dynamics changed, respectively.

Sov Zdravookhr, 1991, (9), 35 - 8
{Prevalence of large-intestinal diseases in Azerbaijan}; Salimov TG et al.; 15,000 adults in Azerbaijan have been surveyed . The programme of the survey included interviews with specially designed questionnaires consisting of questions related to working and living conditions, nutritional habits, X-ray and endoscopic examinations of the colon have been conducted . Referrals to polyclinics for colon diseases accounted for 6.0 per 100 inhabitants and after active detection among men--41.5 and among women--45.6 per 100 persons examined . The leading role in the incidence of chronic diseases of colon is played by hemorrhoid (31.0%), chronic colitis (33.6%), colon dyskinesia (13.5%) . It has been found that the most significant predisposing causes of colon diseases were: bacillary dysentery, different types of neuroses, milk indigestion, gynaecological and prostatic diseases, and obesity.

Probl Tuberk, 1991, (8), 14 - 6
{Pulmonary tuberculosis among the residents of the earthquake area in the Armenian SSR}; Karapetian ET et al.; The examination of 300 patients from the zones of disaster conducted in the hospital, Republican Clinical Antituberculosis Dispensary, has shown an increased number of patients with newly diagnosed pulmonary tuberculosis and a high incidence of exacerbations of the chronic forms of the specific process . It also has revealed the development of disseminated forms with massive bacillary excretion, a higher incidence of various tuberculosis complications, combination of pulmonary tuberculosis and diabetes mellitus and reduction of the body's defensive forces . Low effectiveness of treatment and deterioration of the epidemiological situation in the Republic were due to the above factors and short hospital stay of patients.

Mikrobiologiia, 1991 Jan-Feb, 60(1), 93 - 100
{Screening of Bacillus proteases with plasmin and activator types of action on fibrin}; Vybornykh SN et al.; Forty cultures of aerobic sporogenic bacteria were screened for the plasmin and activator action of bacillar proteases on fibrin . The microorganisms were grown in chemically defined and complex potato-peptone media . Some active cultures were found to produce enzymes with both plasmin and activator types of action . The following organisms exerted the highest activity of the plasmin type; Bacillus macerans PI, strain P3, B . licheniformis 125 and B . firmus . The most active enzymes with the activator type of action were produced by B . macerans PI, Bacillus sp . 21P-7-1, strain 21P-7-2 and B . licheniformis 125.

Probl Tuberk, 1991, (3), 16 - 9
{Specific prevention of tuberculosis in young patients by BCG vaccine}; Bubochkin BP et al.; The efficacy of antituberculous BCG revaccination studied in workers, students and young subjects of the penitentiary labour establishments (PLE) was assessed by the level of tuberculin sensitivity, the presence of a postvaccination skin sign, tuberculosis morbidity and severity of a tuberculosis process . The highest efficacy was achieved in workers and students as a result of systematic revaccination . Tuberculosis morbidity in them was 26.2 and 34.5 per 100,000, respectively . In the PLE contingent it was several times higher because they received revaccination as early as their school years . The postvaccination skin signs were found in 91.8% of workers, 76.4% students and 60.5% young subjects of PLE . In students and workers reactions with an infiltrate size of 5-11 mm were predominant and in the PLE contingent those with an infiltrate of 12 mm and more . Subjects having a skin sign had more benign tuberculosis course . Bacillary excretion was found in 48.9% and in those without a skin sign in 72% of cases (p less than 0.05).

Cancer Immunol Immunother, 1991, 33(3), 189 - 97
Major histocompatibility complex class II antigen expression on leucocyte subpopulations in the draining lymph node and tumour in the early phase of bacillus-Calmette-Guérin-induced tumour regression; Steerenberg PA et al.; We investigated the cellular composition and the major histocompatibility complex (MHC) class II antigen expression in the draining lymph node and the tumour during potentiation of the immune response by intralesional bacillus Calmette-Guerin (BCG) administration in the line 10 hepatocellular carcinoma in the strain 2 guinea-pig . Five days after its injection BCG induced a ninefold increase in the number of draining lymph node cells and an increased MHC class II expression . This increased MHC class II expression was mostly due to the selective increase of B cells in the lymph nodes, and to a lesser extent to the increase of T cells expressing MHC class II antigens . Taking into account this nine-fold increase, intralesional treatment of BCG increased considerably the number of T helper/inducer (anti-CT7) and T suppressor/cytotoxic (anti-CT6) lymph node cells expressing MHC class II antigen . The percentage of tumour-infiltrating T cells expressing MHC class II antigen in the tumour was higher than the percentage of T cells in the regional draining lymph node of non-treated guinea-pigs, indicating the presence of activated T cells in the tumour . After treatment with BCG no further increase in MHC class II expression was measured in the tumour, nor was any phenotypical change of the tumour-infiltrating T cells found . In conclusion, with the use of two-colour flow cytofluorometry we have shown that the potentiation of the already existing immune response to line 10 is accompanied by a considerable increase in T helper/inducer, T suppressor/cytotoxic cells and MHC class II antigen in the regional lymph node . Whether this is essential for the potentiation of the immune response causing tumour regression and long-lasting immunity is a subject for further study.

Bol Oficina Sanit Panam, 1991 Jan, 110(1), 26 - 32
{Protective effect of intradermal BCG on tuberculous meningitis}; Costa Mda C et al.; The incidence of tuberculosis in Bahia, though declining, remains very high, and one of the severe forms of the disease is tubercular meningitis . This case-control study sought to compare confirmed cases of tubercular meningitis in children 0 to 14 years of age with members of a control group, matched as closely as possible for age (not more than about six months apart), who had shown no sign of meningitis or neurological disease . The comparison considered such factors as schooling of the parents, their occupations, exposure to the bacillus at home, and others . The study showed that failure to vaccinate heightens the risk of contracting the disease--a risk estimated at 11.7 with a confidence interval of 4.5 to 30.5, and an etiological component of 9.14%--and also that the risk is lower the earlier the child is vaccinated . With this study the authors wish to underscore the need to expand the coverage of intradermal BCG vaccination.

J Pharm Biomed Anal, 1991, 9(10-12), 843 - 7
Separation and isolation of the isomers of bacitracin by high-performance liquid chromatography and their relationship to microbial activity; Bell RG; Bacitracin, a polypeptide antibiotic produced from strains of Bacillus licheniformis, is one of the most commonly used antibiotics in the world . Actually, the various products generally referred to as 'bacitracin' are mixtures of similar polypeptides which may differ by only one amino acid . The approved method of analysis for bacitracin is microbial . To correlate the microbiological method with an HPLC method, bacitracin was chromatographed using a YMC basic column with UV detection . Adequate separation of the isomers were obtained to scale up this procedure to preparative HPLC using a 250 x 21 mm YMC basic column . The various fractions were separated, isolated and examined for microbial activity . The chromatograms can accurately predict in minutes the microbiologically-determined potency which usually takes 16-24 h to develop . The chromatographic procedure also provides information on the amounts of isomers and degradation products present in the sample, whereas the microbiological assay only provides activities or potencies of the antibiotic . The reported HPLC method also possesses some advantages over some other published HPLC methods in terms of accuracy and time of analysis.

Wiad Parazytol, 1991, 37(3), 357 - 65
{Insecticidal activity of various strains of Bacillus against larvae of houseflies (Musca domestica)}; Lonc E et al.; Insecticidal activity of B . thuringiensis subsp . morrisoni and B . thuringiensis subsp . darmstadiensis as well as B . sphaericus 2362 and 2297 was evaluated against larvae of the house fly (Musca domestica) under laboratory condition . The activity of all tested strains was similar, LC50 ranges from 4 x 10(7) to 1 x 10(8) spores/ml . The higher spores concentration (2 x 10(9)) caused ca 70 per cent mortality . The relative low sensitivity of house fly larvae could results from weakly alkalic pH of the gut (7.6-7.8).

Am J Nephrol, 1991, 11(6), 501 - 4
Renal complications of intravesical bacillus Calmette-Guérin therapy; Modesto A et al.; We report 3 patients who developed signs of systemic infection and renal insufficiency after intravesical bacillus Calmette-Guerin (BCG) instillations for a bladder cancer . Renal biopsy showed tubulo-interstitial nephritis with or without epithelioid granulomas in 2 cases, and mesangial glomerulonephritis in the last case . All patients had granulomatous hepatitis in association . It seems that hematogenous dissemination via a traumatic instillation of BCG and/or an immune-complex mechanism may have contributed to the renal damage, which was only partially reversible in 2 patients.

Acta Urol Belg, 1991, 59(4), 149 - 60
Evolution of the urothelial bladder field after intravesical Bacille Calmette-Guérin therapy; Geavlete P et al.; After transurethral resection (T.U.R) and open surgery of 178 patients with superficial bladder tumors (pTa, pT1) we applied intravesical Romanian (modified Pasteur strain) Bacille Calmette-Guerin therapy . An introduction phase consisting of 8 weekly instillations was followed by a maintenance phase of instillations monthly for 10 months and then 3 monthly for 2 years . Two cases with primary Carcinoma in situ (Cis) were treated with the same protocol . The random urothelial biopsies (4 quadrant) showed before immunotherapy 105 cases (58.33%) with simple (36 cases), medium (36 cases), severe (13 cases) dysplasia and Cis (20 cases) and after 3 years of treatment we found 37 cases (20.55%) with simple (14 cases), medium (14 cases), severe (9 cases) dysplasia and two cases with invasive stage after Cis . The response rate was 75, 55% at 3 years . The mean period of follow up was 50 months . The complications of this therapy were frequent but mild . These results demonstrated the dynamic evolution of the urothelial bladder field after "long-course treatment" with intravesical BCG.

Rev Argent Microbiol, 1991 Jan-Feb, 23(1), 26 - 9
{Enrichment for mutants of Bacillus megaterium deficient in the synthesis of poly-beta-hydroxybutyrate (PHB)}; Floccari ME et al.; Centrifugation through sucrose gradients was adapted to separate spore-forming cells of B . megaterium deficient in poly-beta-hydroxybutyrate synthesis from wild type cells . The conditions described allowed the detection of mutant clones screening a low percentage of the mutagenized population.

Probl Tuberk, 1991, (12), 27 - 8
{Clinical significance of drug resistance of tuberculosis mycobacteria and L-forms}; Golanov VS et al.; The clinical features of a tuberculous process were studied in 101 patients with active pulmonary tuberculosis among whom 76% of the patients lost their social adaptation . The patients were divided into 2 groups: group 1 comprising 61 (60.4%) subjects who excreted tuberculosis mycobacteria and L-forms and group 2 consisting of 40 (39.6%) who excreted only tuberculosis mycobacteria . The frequency and degree of drug resistance were more manifest in 63.8% of the patients of group 1 and in 40% of those of group 2 (p less than 0.01) . Drug resistance to relatively new antituberculosis drugs (ethambutol and rifampicin) was found in 30% of the patients . Mixed bacillary population was found 2 times more frequently in patients with serious progressive forms of tuberculosis, which gives evidence to consider it as a prognostically unfavourable indicator.

Probl Tuberk, 1991, (12), 24 - 6
{Intravenous chemotherapy and galvanization of damaged lung regions in patients with newly diagnosed pulmonary tuberculosis}; Strelis AK et al.; The results of a combined treatment of 179 patients with newly diagnosed destructive pulmonary tuberculosis are presented . Patients of the main group (89 subjects) were given intermittent intravenous chemotherapy with simultaneous galvanization of the affected pulmonary zone, while patients of the control group (90 subjects) received the same treatment but without galvanization . In pulmonary tuberculosis patients who had undergone intracutaneous electrophoresis, the body temperature normalized significantly more rapidly (within 2.2 +/- 0.2 weeks), weakness and weakness disappeared (within 1.2 +/- 0.2 months) and cough ceased (1.6 +/- 0.2 months); sputum expectoration disappeared or substantially reduced (within 1.5 +/- 0.1 months); leukocytosis came to an end (within 1.5 +/- 0.1 months) . The basic parameters of a spirogram improved in a shorter period and to a greater degree . Bacillary excretion ceased more rapidly (within 2.2 +/- 0.4 months), so did the infiltrative phenomena resolute (within 3.0 +/- 0.1 months) and pulmonary destructive changes disappear (within 3.3 +/- 0.2 months).

Roum Arch Microbiol Immunol, 1991 Jan-Mar, 50(1), 61 - 6
Long-term effects of Bacillus thuringiensis subsp . israelensis on Aedes aegypti; Ceianu C; Immediate (24 hours exposure) and long-term (until the emergence of the adults) effects of different doses of a primary powder of Bacillus thuringiensis subsp . israelensis (B.t.i.) against first and second instar Aedes aegypti larvae were monitored . The long-term effect was dose-dependent and was materialized by a prolongation of the preimaginal development and continuous cumulative mortality until the emergence of the adults . Mortality values were higher during the fourth larval instar and the pupal stage . Some of the larvae reaching the fourth instar were smaller in size and remained in this state a 2-4 times longer period than in the control and finally died as larvae or very small pupae . The long-term effect was more intense as the treatment was applied earlier during the larval development . The correlation of the immediate lethal effect with the late effect allows the evaluation of the total impact of a larvicidal treatment.

Rom J Morphol Embryol, 1991 Jan-Jun, 37(1-2), 61 - 8
Urothelial bladder biopsy--important evolution prognosis factor of superficial bladder cancer under intravesical Bacille Calmette-Guérin therapy; Geavlete P et al.; During 11 years we investigated the value of multiple randomised biopsies (4 quadrant) and by in vivo staining with methylene blue (chromocystoscopy) of 231 cases with superficial bladder tumours under complementary intravesical Bacille Calmette-Guerin therapy (3 years of immunotherapic treatment after our protocol) . We discovered the appearance of 20 cases with carcinoma insitu (Cis) and 105 cases with different degrees of dysplasia (simple, medium and severe) . We demonstrated 69.44% pathological bladder biopsies on normal-looking mucosa . 75% of the recurrent tumours (44 cases-22.44%) had pathological bladder urothel al field modifications associated with bladder cancer . On this occasion, we point out the highest prognostic value of the complex protocol of superficial bladder tumour evaluation based on the study of the bladder biopsies as an essential parameter.

Rev Cubana Med Trop, 1991 Jan-Mar, 43(1), 39 - 44
{The pilot project results of applications of the biolarvicide Bacillus sphaericus 2362 on mosquito breeding grounds of the town of Santa Cruz del Norte (La Habana Province)}; Montero Lago G et al.; The effectiveness of the Bacillus sphaericus strain 2362 in liquid formulation, at a 10 ml/m2 dose, was tested in 157 breeding sites of Culex quinquefasciatus mosquitos, 2 breeding sites of Culex quinquefasciatus and Anopheles albimanus and 1 breeding site of Aedes taeniorhynchus, consisting of 1 river, 2 oxidation ponds, 1 pond, 4 dams, 2 microdams and 150 pits . The results reached with the application of 1800 1 of biolarvicide show its effectiveness . Within 24 hours of treatment, mortality rates were 100%, in a wide range of ecological conditions, its permanence up to 5 months in breeding sites without stream and innocuousness for other hydrobionts . DPGs in humans, having shown considerable reductions, confirm the advantages and potentialities of using this method under tropical conditions.

J Egypt Public Health Assoc, 1991, 66(3-4), 333 - 44
Cloning and expression of the lepidopteran toxin produced by Bacillus thuringiensis var Sotto in Escherichia coli; Rady MH; During sporulation, Bacillus thuringiensis var . Sotto produces a parasporal crystalline protein which is toxic for the silk-worm, Bombyx mori and the cotton leaf worm, Spodoptera littoralis . The gene coding this crystal protein is present in a single plasmid . The plasmid DNA was isolated, purified and physically mapped using restriction endonuclease enzymes (R.E.) . The gene coding the delta-endotoxin was inserted into Escherichia coli-Jm103, using cloning vector pUC8 . Transformed E . coli cells were found to synthesize the delta-endotoxin as demonstrated by the pathogenicity of the transformed cells against 4th instar larvae of S . littoralis.

Rocz Panstw Zakl Hig, 1991, 42(1), 71 - 4
{Effect of Escherichia coli on growth of Bacillus cereus}; Stec E; The effect of 101 Escherichia coli strains on growth of 90 Bacillus cereus strains on solid media was investigated . Only 9 E . coli strains (in particular the colicin +-generating ones) were antagonistic towards B . cereus, giving distinct growth-inhibition zones around the colonies.

Microbiol Immunol, 1991, 35(7), 557 - 67
Lysis of murine macrophages by lymphokine-activated killer-like cells induced by BCG in vitro; Chen MF et al.; The lymphokine-activated killer (LAK)-like activity was found to be induced in mouse splenocytes cultured together with Bacillus Calmette-Guerin (BCG) . The killer cells induced by BCG were capable of killing both NK-sensitive (YAC-1, P388D1) and NK-resistant (P815) tumor cells . As an important finding, they also lysed syngeneic macrophages (M phi) . The anti-M phi killer activity appeared on day 2, and reached a peak on day 5 of culture . Phenotype analysis of the killer cells by depletion techniques using monoclonal antibody (mAb) and complement indicated that the majority of these anti-M phi killer cells were Thy-1+ and asialo GM1+ . This M phi cytolysis could be inhibited by the addition of cold M phi, YAC-1 tumor cells, and P815 tumor cells, suggesting that the same population of the effector cells recognize M phi and tumor cells . The addition of anti-MHC class I, anti-MHC class II, anti-L3T4, or anti-Ly-2 mAb directly to assay cultures did not affect anti-M phi cytolysis, suggesting that the MHC molecules are not involved in the cytolysis of M phi by the BCG-induced killer cells . The addition of anti-LFA-1 mAb partially inhibited the cytotoxicity, suggesting importance of the contact between targets and effectors in the cytolysis . Our present data suggest that activation of murine lymphocytes with BCG induces LAK-like cells capable of killing a wide variety of tumor cells as well as M phi and this anti-M phi cytolysis is mediated by nonspecific killer cells.

Urol Int, 1991, 47(4), 250 - 4
Combined effect of interleukin 2 and bacillus Calmette-Guérin in the therapy of mice with transitional cell carcinoma; Ikemoto S et al.; We investigated the effect of combination therapy with interleukin 2 (IL-2) and Bacillus Calmette-Guerin (BCG) on C3H/HeN mice implanted with mouse bladder tumor cells (MBT2) . MBT2-bearing mice were treated with a local injection of BCG into the tumor at a dose of 1 mg/day for a total of 3 times and/or a 10-day continuous subcutaneous infusion of 5 x 10(4) units IL-2/day from day 11 to day 20 . As a result, the growth of the tumor in mice treated with IL-2 alone was slightly suppressed, while tumor growth was hardly suppressed in mice treated with BCG alone . However, when IL-2 was administered with BCG, tumor growth was strongly suppressed and mean survival time was prolonged . Natural killer cell activity in the spleen cells was most elevated in mice treated with IL-2 and BCG, while lymphokine-activated killer cell activity was not observed in all groups . Lymphocyte subset analysis showed that there was little change in the ratio of Lyt2-positive lymphocytes or that of L3T4-positive lymphocytes . These findings suggested that clinical evaluation of combination therapy with IL-2 and BCG may be worthwhile.

Probl Tuberk, 1991, (6), 10 - 2
{Tuberculosis morbidity in high-risk groups of children}; Kozlova AV et al.; A follow-up of 2052 children has shown that the level of Mycobacterium tuberculosis contamination among the subjects frequently getting ill with ARVD is 1.5-2 times higher than among healthy ones . This parameter was 1.5-2 times higher among pyelonephritis patients than among healthy subjects . Hyperergic tuberculin reactions more often developed in children being in tuberculosis infection foci, including 58.3% from the bacillary environment . A close association of tuberculin hypersensitivity and the action of aggravating factors is often accompanied by immunity derangement . According to our data, even sufficiently adequate BCG vaccination fails to prevent local tuberculosis which develops in 1.9%.

Probl Tuberk, 1991, (11), 67 - 9
{Antituberculosis antibodies detected by an immunoenzyme test in patients with pulmonary tuberculosis}; Litvinov VI et al.; The method of indirect solid-phase enzyme immunoassay (EIA) was used to detect antibodies in the sera of 166 pulmonary tuberculosis patients and 56 healthy donors . A preparation with a mol . mass of 38-42 kD was used as an antigen which was isolated from the mycobacteria H37Rv by a consecutive separation under high pressure, extraction of KCl cellular membranes and gel-filtration in the gel Toyopearl HW 55F . Antituberculous antibodies (AtAb) were detected by the EIA method in 94% of pulmonary tuberculosis patients which was much higher as compared to the same parameter in healthy subjects (10.7%) . Hence, AtAb detection by this method can serve as an additional criterion for tuberculosis diagnosis . The detection rate and AtAb level are higher in fibrocavernous tuberculosis than those in infiltrative tuberculosis . The AtAb detection rate is higher in manifested intoxication than in moderate one or its absence . AtAb are more often detected in chronic than in newly diagnosed tuberculosis, in the disseminated forms than in the limited forms, in pronounced infiltration in the lungs as compared to a moderate form, and also in patients with bacillary excretion than in those whose sputum had no M . tuberculosis.

DNA Seq, 1991, 1(3), 173 - 80
Sequencing with the large fragment of DNA polymerase I from Bacillus stearothermophilus; McClary J et al.; The large fragment of DNA polymerase I, isolated from Bacillus stearothermophilus, was used for dideoxy sequencing . This heat-stable enzyme permits performing sequencing reactions at high temperature to melt secondary structure and results in uniform band intensities and low background on the autoradiogram . The enzyme can be used in the standard Sanger one-step protocol or in a two-step protocol which separates the labeling reaction from the elongation-termination reaction . The enzyme can be used in double-stranded sequencing . 35S-labeled nucleotides may be used instead of 32P-labeled nucleotides . Both 7-deaza-dGTP and dITP can be used during the reaction in order to minimize band compression on the gel . Results presented here indicate that this enzyme should be a useful tool for sequence determination.

Chin J Biotechnol, 1991, 7(1), 63 - 72
Cloning and expression of penicillin G acylase gene in Bacillus megaterium; Zhang LF et al.; Bacillus megaterium BM1, which produces penicillin G acylase (PGA), has been isolated . Gene encoding for PGA was cloned into E . coli MC1061 using pBR322 as the vector, obtaining a recombinant plasmid pBmPA4 containing 9.9 kb inserted DNA . Restriction map of the plasmid was analyzed . A pBmPA5 containing 4.9 kb was gained by deletion in vitro . Both pBmPA4 and pBmPA5 clones can be expressed in E.coli MC1061, and their expressions were induced by phenylacetic acid.

Dermatologica, 1991, 183(1), 25 - 30
Adjuvant treatment in stage I and II malignant melanoma: a randomized trial between chemoimmunotherapy and immunotherapy; Castel T et al.; A randomized trial comparing chemoimmunotherapy (bacillus Calmette-Guerin + chemotherapy) and immunotherapy (bacillus Calmette-Guerin alone) was carried out in high-risk stage I and II malignant melanoma patients . Eight-two evaluable patients were included . The follow-up median duration was 11 years . Recurrent melanoma developed in 28 patients (34%) . The overall survival rate was 76% at 5 years and 65% at 10 years . There were no statistical differences in survival probability or disease-free survival (DFS) probability between the two treatment groups . Survival and DFS were also analyzed in relation to various prognostic factors . Statistically significant differences were only seen in a subset of 33 patients with lower limb malignant melanoma, the prognosis being better for the chemoimmunotherapy group in this location . Because of the small number of patients in each group, a week positive effect of either of these two treatments cannot be ruled out . Chemoimmunotherapy only seems to improve the prognosis of stage I or II malignant melanoma of the legs.

Nephrologie, 1991, 12(5), 233 - 6
{Vertebral tuberculosis in a chronic hemodialysis patient with polycystic kidney disease}; Chawki M et al.; We report a case of vertebral tuberculosis in a female patient with polycystic kidney disease (PKD) who had been treated with maintenance hemodialysis for six years . Diagnosis of lumbar bacillary spondylitis was particularly difficult, owing to proximity of polycystic kidneys and spine . After removal of one kidney in which no bacterial focus was found, pain and inflammation persisted . Two months later CT scan and RMN evidence of 4th lumbar vertebral lysis led to performing open vertebral biopsy . Tuberculous lesions were found on this material and antibacillary treatment was begun . This case illustrates the difficulty of diagnosing vertebral tuberculosis in dialysis patients, especially in those with PKD.

Probl Tuberk, 1991, (8), 36 - 8
{Effectiveness of the treatment of patients with fibrous-cavernous pulmonary tuberculosis}; Sadykov AS; The follow-up of 301 patients with fibrous-cavernous pulmonary tuberculosis has demonstrated that the disease outcome and treatment effectiveness depend on the nature of its course . A long-term specific therapy for 9-10 months brings about cessation of bacillary excretion in 50-60% of cases even in a progressive course of the disease and closure of destruction cavities occurred in 12.5% of cases with rapid progression and in 21.7% of cases with slow progression of the disease . Despite all measures taken, stabilization of the process fails in one-fourth of the patients with rapidly progressing fibrous-cavernous tuberculosis and in 9% of those with a slowly progressing course . In 90% of cases bacillary excretion in patients with stable fibrous-cavernous tuberculosis ceased in 3 months and in 100% after 6 months of treatment.

Cancer Immunol Immunother, 1991, 34(3), 186 - 90
Immunotherapy of bovine ocular squamous cell carcinoma by repeated intralesional injections of live bacillus Calmette-Guérin (BCG) or BCG cell walls; Rutten VP et al.; Thirty cows of the Dutch Friesian and the Maas-Rijn-Ijssel breed with histologically confirmed ocular squamous cell carcinoma were treated by repeated intralesional injection of live bacillus Calmette-Guerin (BCG) (n = 14) or a BCG cell-wall vaccine (n = 16) . Complete regression of the primary tumour was observed in 64% and 57% of the animals respectively . In the 2-year follow-up period there was no recurrence of primary tumours . This sharply contrasts with the recurrence frequency (40%-50%) after complete remission induced by a single intralesional injection with BCG, observed in an earlier study . In 1 animal a new primary tumour developed . At necropsy metastases were present in 33% of the treated animals: in 3 of 17 animals that showed complete regression of the primary tumour and in 7 of 13 animals with partial regression or progressive disease . This did not differ significantly from results obtained after a single treatment (27%) . Delayed-type hypersensitivity to M . bovis purified protein derivative (PPD) was more persistent in animals showing regression of the primary tumour than in non-responding animals . Of the animals with a positive PPD response 6 months after treatment, 79% showed tumour regression . Regression was observed in only 28% of the animals not responding to PPD after the same period of time . In conclusion: (a) recurrence of the primary tumour was not observed after repeated BCG treatment; (b) the frequency of metastases was not decreased compared to results obtained with a single treatment; (c) regression was correlated with a positive delayed-type hypersensitivity reaction to PPD (P less than 0.05) 6 months after treatment; (d) no significant differences were observed when the clinical results of treatment with live BCG and the BCG cell wall vaccine were compared.

Eur Urol, 1991, 20(2), 170 - 2
Treatment of carcinoma in situ of the urethra with intraurethral instillations of bacillus Calmette-Guérin . Case report and review of literature; Witjes JA et al.; We present the case of a 60-year-old man with carcinoma in situ of the urethra after previous successful radiotherapy of an invasive bladder tumor . Intraurethral instillations with bacillus Calmette-Guerin were given . In spite of initial success, recurrences developed after two courses of 12 intraurethral instillations, and radical surgery was performed . The use of bacillus Calmette-Guerin is reviewed with regard to the locations of superficial transitional cell carcinoma outside the bladder cavity.

Eur Urol, 1991, 20(1), 19 - 25
Intravesical chemotherapy (mitomycin C) versus immunotherapy (bacillus Calmette-Guérin) in superficial bladder cancer; Rintala E et al.; Both intravesical mitomycin C (MMC) and bacillus Calmette-Guerin (BCG; Pasteur strain F) were effective in the present prospective randomized multicenter study consisting of 91 patients with frequently recurrent superficial (Ta-T1) bladder cancer . The result was in favour of BCG, as shown by the measurements with complete response (CR), disease-free interval and recurrence rate . CR of 58% with MMC and 40% with BCG were reached in 22 instillation series on carcinoma in situ of 18 patients . Due to side effects, MMC instillations were discontinued in 8.6%, and BCG instillations in 19.6%, respectively . After the 2-year follow-up also 1 case of pulmonary tuberculosis occurred in the BCG group.

Acta Microbiol Pol, 1991, 40(3-4), 235 - 41
The metabolism of anthracene and 9,10-dimethylanthracene by bacteria isolated from waters; Traczewska TM et al.; The metabolism of two polycyclic aromatic hydrocarbons i.e . anthracene and 9,10-dimethylanthracene by Micrococcus sp., Pseudomonas sp . and Bacillus macerans was examined . The above compounds were used as a sole carbon source for their growth . Using the reversed-phase thin layer chromatography techniques a number of anthracene and 9,10-dimethylanthracene metabolites were isolated and their structures identified spectroscopically . These included anthracene and 9,10-dimethylanthracene cis-dihydrodiols, hydroxy-methyl-derivatives and various phenolic compounds . Bacteria metabolise hydrocarbons using the dioxygenase enzyme system, which differs from the mammalian cytochrome P-450 monoxygenase . Hence in addition rat liver microsomal metabolism of the above hydrocarbons was investigated using the same separation techniques.

Am J Gastroenterol, 1991 Jan, 86(1), 79 - 81
Whipple's disease can mimic chronic AIDS enteropathy; Maliha GM et al.; Previous case reports have demonstrated that the intestinal pathology of Mycobacterium avium-intracellulare (MAI) infection in the acquired immune deficiency syndrome (AIDS) has a light microscopic appearance similar to Whipple's disease . This case report describes a 52-yr-old male patient with a clinical picture suggestive of AIDS, including diarrhea, weight loss, oral thrush, and intestinal cryptosporidiosis . The intestinal biopsy showed light microscopic features compatible with either MAI or Whipple's disease, but electron microscopy confirmed the presence of the Whipple bacillus . Markers of human immunodeficiency virus (HIV) infection were absent . Although immune abnormalities have been reported in Whipple's disease, this is the first report of opportunistic infections complicating this condition . A useful clinical pearl emerges from this and other cases: AIDS can mimic Whipple's disease; Whipple's disease can mimic AIDS.

Virchows Arch B Cell Pathol Incl Mol Pathol, 1991, 61(3), 159 - 67
Intravesical BCG administration in the guinea pig . II . Immunohistochemical characterization of cellular infiltrate after one or two cycles of intravesical BCG; de Jong WH et al.; The intravesical administration of bacillus Calmette-Guerin (BCG) is used as an adjuvant therapy after transurethral resection for superficial bladder cancer in man . The aim of this study was to characterize with monoclonal antibodies the cellular infiltrates in the bladder wall occurring after one or two cycles, each comprising six weekly intravesical BCG administrations in guinea pigs . BCG-RIVM and BCG-TICE were instilled once a week in undamaged bladders at weeks 1-6 and 11-16, respectively, and retained in the bladder for 1 h . Autopsy was performed 1 week after the last BCG administration at weeks 7 and 17 . By routine histology, a single BCG course induced scattered focal infiltrates consisting of minor to severe accumulations of mononuclear cells . After two cycles of BCG, a much more severe inflammatory reaction sometimes occupying almost the total bladder mucosa was noted . The major component of this inflammation was mononuclear cells, mainly lymphocytes, while typical BCG granulomas were occasionally observed . In general, the epithelial layer of the bladder showed no visible alterations . Immunohistochemical staining showed that the majority of mononuclear cells present in the infiltrate were T-cells . Widely scattered B-cells and macrophages were also present, but were much fewer in number than the T-cells . In general, T-helper/inducer cells were more frequent than T-suppressor/cytotoxic cells . Pseudofollicles, also noted after H&E staining, consisted mainly of B-cells . The second series of BCG administrations produced pseudofollicles in the infiltrates in all animals investigated.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbiol Immunol, 1991, 35(3), 223 - 34
Detection of Bacillus cereus flagellar antigen by enzyme-linked immunosorbent assay (ELISA)
Murakami T, Hiraoka K, Mikami T, Matsumoto T, Katagiri S, Suzuki M.
A serological typing scheme of Bacillus cereus has been developed by immunochemical analyses of flagellar antigen using an agglutination method . Enzyme-linked immunosorbent assay (ELISA) for the classification of flagellar serotype of Bacillus cereus had greater sensitivity . 10-500 times, than that of agglutination method . The specificity of flagellar antigen and antibody was determined by immunogold electron microscopy and ELISA inhibition assay . Application of ELISA is useful for the detection of the small amounts and many kinds of antigen-antibody reactions.

Toxicon, 1991, 29(3), 386 - 90
Human immunosuppression induced by sea nettle (Chrysaora quinquecirrha) venom; Wachsman M et al.; Large intradermal injections of crude sea nettle (Chrysaora quinquecirrha) venom in normal saline produced immunosuppression in a healthy human adult male . This response persisted several days and was homologous against that coelenterate antigen but also heterologous against antigens contained within vaccinia and herpes simplex viruses and tetanus bacillus . This down-regulation of immunity was probably mediated by cells with functional properties of suppressor cells and could be reversed by indomethacin suggesting a role for prostaglandins . This result suggests the possibility that naturally occurring environmental agents, other than sunlight, may influence the human immune response.

Plant Foods Hum Nutr, 1991 Jan, 41(1), 27 - 34
Comparative study of microorganisms and sensory attributes of condiments from the fermentation of different seeds; Jideani IA et al.; Seeds of African locust bean, melon, castor oil bean and soybean were processed and fermented for 3 days to produce local condiments in the laboratory with a method that simulated the traditional production process . Microorganisms associated with their fermentation and the organoleptic properties of the products were compared . Altogether, seven species of bacteria were involved in the fermentation . These included five Bacillus spp . and one species each of Pseudomonas and Staphylococcus . Their occurrence vary between the seeds on the different days of fermentation . However, Bacillus spp . were present in all the seeds throughout the fermentation period . The sensory evaluation preference rating for the four products was highest for soybean condiment, followed by that made from locust bean . Melon condiment was the least preferred among the four products . Statistical analysis showed that there were significant differences (P less than or equal to 0.05) among the four products for each of the four organoleptic properties evaluated by the judges and also for titratable acidity . Glutamic acid level of soybean condiment was highest (0.31%) among the four products with that of melon being the lowest (0.04%) . These results could serve as useful indices for the development of starter culture and optimization of production process in commercializing the production of these local condiments.

J Bacteriol, 1991 Jan, 173(2), 791 - 800
Sodium ion-dependent amino acid transport in membrane vesicles of Bacillus stearothermophilus; Heyne RI et al.; Amino acid transport in membrane vesicles of Bacillus stearothermophilus was studied . A relatively high concentration of sodium ions is needed for uptake of L-alanine (Kt = 1.0 mM) and L-leucine (Kt = 0.4 mM) . In contrast, the Na(+)-H(+)-L-glutamate transport system has a high affinity for sodium ions (Kt less than 5.5 microM) . Lithium ions, but no other cations tested, can replace sodium ions in neutral amino acid transport . The stimulatory effect of monensin on the steady-state accumulation level of these amino acids and the absence of transport in the presence of nonactin indicate that these amino acids are translocated by a Na+ symport mechanism . This is confirmed by the observation that an artificial delta psi and delta mu Na+/F but not a delta pH can act as a driving force for uptake . The transport system for L-alanine is rather specific . L-Serine, but not L-glycine or other amino acids tested, was found to be a competitive inhibitor of L-alanine uptake . On the other hand, the transport carrier for L-leucine also translocates the amino acids L-isoleucine and L-valine . The initial rates of L-glutamate and L-alanine uptake are strongly dependent on the medium pH . The uptake rates of both amino acids are highest at low external pH (5.5 to 6.0) and decline with increasing pH . The pH allosterically affects the L-glutamate and L-alanine transport systems . The maximal rate of L-glutamate uptake (Vmax) is independent of the external pH between pH 5.5 and 8.5, whereas the affinity constant (Kt) increases with increasing pH . A specific transport system for the basic amino acids L-lysine and L-arginine in the membrane vesicles has also been observed . Transport of these amino acids occurs most likely by a uniport mechanism.

J Enzyme Inhib, 1991, 5(2), 111 - 7
A synthesis of acylphosphonic acids and of 1-aminoalkylphosphonic acids: the action of pyruvate dehydrogenase and lactate dehydrogenase on acetylphosphonic acid; Dixon HB et al.; Acylphosphonic acids, R-CO-PO(OH)2, have been synthesized by the steps {formula: see text} of which the last is new and provides a mild method for de-esterifying acylphosphonic acids . Their reductive amination gives a simple way of making 1-aminoalkylphosphonic acids . Acetylphosphonic acid inhibited NAD+ reduction by pyruvate with the pyruvate dehydrogenases from Escherichia coli and Bacillus stearothermophilus . The inhibition was competitive with pyruvate, with Ki of 6 microM for the E . coli enzyme (pyruvate Km 0.5 mM) and one of 0.4 mM of the B . stearothermophilus enzyme (pyruvate Km 0.1 mM) . Acetylphosphonate and its monomethyl ester are substates for pig heart lactate dehydrogenase, with Km values of 15 mM and 10 mM respectively (pyruvate Km 0.05 mM) and specificity constants one thousandth that for pyruvate.

Pediatrie (Bucur), 1991, 40(1-2), 145 - 52
{Pleural puncture biopsy--a diagnostic method in tuberculous serofibrinous pleurisy in children}; Murgoci G et al.; The biopsy of the parietal pleura with a special needle has become an essential diagnosis method in the adult patients with pleural disease of unknown etiology, and might become a complementary paraclinical method for children also . The certitude diagnosis in tuberculous serous fibrinous pleurisy of the child is laid on evidencing Koch's bacillus at the direct examination or in culture, and on fragments of pleural biopsy puncture, in evidencing the lesions . The study is based on the results obtained in the pleural biopsy puncture made on 6 children . The histologic examination of the pleural fragment showed, in 4 cases of 6, the presence of tuberculous lymphoepithelioid nodules, with central caseous necrosis thus granting certitude to the diagnosis . The pleural biopsy puncture permits an early and certain diagnosis in more than 2/3 of the pleurisies of tuberculous etiology.

Chin J Biotechnol, 1991, 7(1), 15 - 23
Cloning of DNA fragments with promoter function from temperate phage of Bacillus licheniformis; Sheng XY et al.; Phage Blp7 DNA was digested with restriction enzyme and ligated to the restriction enzyme digested vector pTG402 . The ligated mixture was used to transform competent cells of E . coli MC1061 . Plasmid DNA was extracted from pooled transformants and competent cells of B . subtilis were transformed . By selecting yellow colonies upon spraying with catechol solution, 22 clones containing DNA fragments with promoter function were obtained . The promoter activity of 15 clones was determined by the color reaction of catechol-2,3-dioxygenase . The inserted fragment of the most potent promoter was mapped with restriction enzymes . CatO2 ase activity of two clones was measured in cells of B . subtilis of all growth phases and was found to increase rapidly at the end of the log-phase . It is inferred that these two promoters might be recognized by sigma 37.

Chin J Biotechnol, 1991, 7(1), 1 - 13
Insect resistance of transgenic tobacco plants expressing delta-endotoxin gene of Bacillus thuringiensis; Tian YC et al.; The initiative B.thuringiensis delta-endotoxin (Bt toxin) gene clones TH12 and TH48 contain two different classes of homologous genes, the 5.3 kb class and 6.6 kb class, respectively . Bt toxin genes of both classes, modified at the 5'-end and truncated at the 3'-end, can still be expressed to produce the insecticidal, truncated toxin proteins in E . coli . The modified Bt toxin genes were inserted into the plant binary expression vector pBin 437 (a derivative plasmid of pBin 19) and were transferred into tobacco by Ti plasmid-mediated gene transfer system . Southern blot and DNA slot blot analysis indicate that the Bt toxin genes have been integrated into tobacco genome at a copy number of 1 to 5 . Northern blot analysis of polyA+ RNAs from progeny of the transgenic plants revealed that Bt toxin genes of both 5.3 kb and 6.6 kb classes were expressed in transgenic plants, though the transcripts were degraded to RNAs of lower molecular weights . In insecticidal test, 5 plants from the progeny of 5.3 kb class gene-transformed SR1 tobacco plants and 3 plants from those of 6.6 kb class gene-transformed plants were found to be toxic to the testing larvea of H.assulta . In comparison with the control, mortality of the insects fed on transgenic plants reached 40-50% and the growth of the survived insects was remarkably inhibited . These results indicate that the modified Bt genes of the 5.3 kb and 6.6 kb classes were expressed in transgenic plants and could confer on the transgenic plants a new character of insect resistance.

Agric Biol Chem, 1991 Jan, 55(1), 207 - 13
The nucleotide sequences of Bacillus stearothermophilus ribosomal protein S12 and S7 genes: comparison with the str operon of Escherichia coli; Kimura M; The primary structure of the ribosomal protein S7 from Bacillus stearothermophilus (BstS7) was analyzed by a combination of amino acid and DNA sequence analysis . Peptide sequence information was derived from tryptic peptides of BstS7 by manual Edman degradation . The nucleotide sequence of the 1.5-kb SalI fragment from B . stearothermophilus genome, cloned by the Escherichia coli S12 gene (rpsL) as a hybridization probe, was determined . Comparison of deduced amino acid sequence with the corresponding sequences of E . coli ribosomal proteins showed that this fragment contains the genes encoding S12, S7, and the N-terminus of the elongation factor G . Thus, the organization of this gene cluster is same as that in the str operon of E . coli . The amino acid sequence of B . stearothermophilus S12 (BstS12) deduced from the nucleotide sequence information agrees with the published amino acid sequence of BstS12 {M . Kimura and J . Kimura, FEBS Lett., 210, 91 (1987)} . The deduced sequence of B . stearothermophilus S7 (BstS7), together with sequence information of tryptic peptides, showed that protein S7 consists of 155 amino acid residues with a calculated molecular weight of 17933 and has 56% amino acid identity with the E . coli S7 (EcoS7).

Agric Biol Chem, 1991 Jan, 55(1), 189 - 94
Analysis by deletion and site-directed mutagenesis of promoters of the cell wall protein gene operon in Bacillus brevis 47; Adachi T et al.; The cell wall protein gene operon of Bacillus brevis 47 has multiple and tandem promoters . The precise locations of the two major promoters (P2 and P3) of the operon were determined by deletion analysis . This together with results of oligonucleotide-directed mutagenesis of the promoter regions confirmed that the -35 and -10 sequences of the two promoters proposed previously are essential for their promoter activity . A G + C-rich sequence upstream of the -35 region of P2 and an A-rich sequence upstream of the -35 region of P3 facilitate the transcription from P2 and P3, respectively.

Biotechnology (N Y), 1991 Jan, 9(1), 47 - 52
Production of human interleukin-3 using industrial microorganisms; van Leen RW et al.; We expressed a cDNA encoding the multicolony stimulating factor interleukin-3 in a variety of cell types, including bacteria, yeast and mammalian cells . After evaluation of the advantages and disadvantages of each potential system, we designed a production and purification scheme using Bacillus licheniformis . The purification consists of hydrophobic interaction chromatography, two steps of ion exchange chromatography and gel filtration . The purified and formulated product entered clinical trials in November 1989.

J Ind Microbiol, 1991 Jan, 7(1), 45 - 52
Structural analysis of Bacillus licheniformis 86 surfactant; Horowitz S et al.; A tentative structure and composition of a surfactant, BL-86, produced by Bacillus licheniformis 86 is described . The surfactant is a mixture of lipopeptides with the major components ranging in size from 979 to 1091 Da and varying in increments of 14 Da . The variation in molecular weight represents changes in the number of methylene groups in the lipid and/or peptide portion of the surfactant . There are 7 amino acids per molecule . The peptide portion is composed of the following amino acids: glutamic acid or glutamine (glx), aspartic acid or asparagine (asx), valine, leucine, and isoleucine at a ratio of 1.0:1.0:1.4:3.0:0.6, respectively . The leucine is present as both the D and L isomers at a ratio of about 2:1, respectively . Forty percent of the molecules contain L-valine instead of L-isoleucine . The glx and asx are present as a combination of L-glutamic acid and L-asparagine and/or L-glutamine and L-aspartic acid . The N-terminus of the peptide is blocked, most likely by a peptide bond to the lipid portion . An ester carbonyl structure is present, which could be a part of a lactone ring connecting the beta position of the lipid to one of the carbonyl groups in the peptide . The lipid portion is composed of, on average, 8-9 methylene groups, and contains a mixture of linear and branched tails . Results of DCI-MS and FAB-MS analyses, as well as surface tension measurements, of purified BL-86 HPLC fractions support the proposed composition.

Eur J Biochem, 1990 Dec 27, 194(3), 897 - 901
A highly active and oxidation-resistant subtilisin-like enzyme produced by a combination of site-directed mutagenesis and chemical modification; Gron H et al.; The subtilisins are known to be susceptible to chemical oxidation due to the conversion of Met222 into the corresponding sulfoxide . A number of derivatives with resistance towards oxidation have previously been prepared by replacement of this group with the other 19 amino acid residues . Unfortunately, the activities of these enzymes were of the order of 1-10% of that obtained with the wild-type enzyme . In contrast, the oxidation-labile cysteine mutant exhibited much higher activity, suggesting that this is associated with the presence of a sulphur atom in the amino acid at position 222 . It is shown here that it is possible to maintain a sulphur atom in the amino acid at position 222 without the enzyme becoming labile towards oxidation . A subtilisin from Bacillus lentus, subtilisin 309, in which Met222 was replaced with a cysteinyl residue by site-directed mutagenesis was modified with thioalkylating reagents . Treatment of such enzyme derivatives with H2O2 revealed that their stabilities towards oxidation had increased significantly compared to both wild-type and unmodified {Cys222}subtilisin . One of the chemically modified enzyme derivatives, {Me-S-Cys222}subtilisin, exhibited a kcat/Km value of 56% of that obtained with the wild-type enzyme when assayed against the substrate Suc-Ala-Ala-Pro-Phe-NH-Ph-NO2 (Suc, succinyl) and it exhibited 89% activity when tested in an assay with dimethyl casein as a substrate . The corresponding values obtained for unmodified {Cys222}subtilisin were lower, i.e . 39% for the dimethyl casein activity and 46% for the kcat/Km for the hydrolysis of Suc-Ala-Ala-Pro-Phe-NH-Ph-NO2 . This demonstrates the feasibility of replacing the oxidation-labile methionyl residue group in a subtilisin enzyme with a group stable towards oxidation without substantially reducing the activity.

Cancer Res, 1990 Dec 15, 50(24), 7750 - 3
Inhibition of antibody response to Pseudomonas exotoxin and an immunotoxin containing Pseudomonas exotoxin by 15-deoxyspergualin in mice; Pai LH et al.; Immunotoxins are potent cell-killing agents that may be useful in the treatment of cancer . The early production of neutralizing antibodies to immunotoxins is one of the major limiting factors for their use in humans . 15-Deoxyspergualin (DSG), a derivative of spergualin, which is a metabolite of Bacillus laterosporus, has been found to have immunosuppressive activity in rodents, dogs, and primates . We examined the suppressive activity of DSG on the antibody response to Pseudomonas exotoxin in mice by enzyme-linked immunosorbent assay . Male BDF1 mice were immunized with a single dose of a nontoxic mutant of Pseudomonas exotoxin (40 micrograms) and then treated with i.p . injections of DSF at a dose of 10 mg/kg for 3 days . Although antibodies to Pseudomonas exotoxin were observed within 7 days in the control group, there was complete suppression of antibody production in the DSG-treated group . Immunosuppression has also been observed in animals immunized with multiple doses (10 mg x 7 d) of Pseudomonas exotoxin and treated with DSG at a dose of 5 mg/kg for 21 days . Similar immunosuppression was observed in mice given multiple doses of the immunotoxin, anti-Tac-LysPE40 . We conclude that the immunosuppressive activity of DSG may be useful in increasing the duration of immunotoxin treatment.

Am J Ophthalmol, 1990 Dec 15, 110(6), 683 - 7
Antibiotic therapy for Bacillus species infections; Kervick GN et al.; We reviewed 36 cases of culture-proven Bacillus species ocular infections occurring between September 1974 and December 1989 . Kirby-Bauer disk sensitivities were available in 34 of the 36 cases (95%) . All Bacillus species isolates were sensitive to the aminoglycoside antibiotics (N = 34) and to vancomycin hydrochloride (N = 32); resistance to clindamycin was found in four of 18 (22%) of tested isolates . Although B . cereus was uniformly sensitive to these antibiotics, resistance to clindamycin occurred in four cases in the non-B . cereus group . The microbroth dilution technique confirmed the Kirby-Bauer data . The aminoglycosides were uniformly effective, but the cephalosporins (first, second, and third generation drugs) were consistently ineffective against B . cereus and varied from sensitive to moderately sensitive for the non-B . cereus isolates . Our microbiologic laboratory findings suggest that vancomycin hydrochloride in combination with an aminoglycoside ensures more consistent antibiotic coverage of Bacillus species ocular infections.

Biochem J, 1990 Dec 15, 272(3), 613 - 9
Site-directed mutagenesis of beta-lactamase I . Single and double mutants of Glu-166 and Lys-73; Gibson RM et al.; Two single mutants and the corresponding double mutant of beta-lactamase I from Bacillus cereus 569/H were constructed and their kinetics investigated . The mutants have Lys-73 replaced by arginine (K73R), or Glu-166 replaced by aspartic acid (E166D), or both (K73R + E166D) . All four rate constants in the acyl-enzyme mechanism were determined for the E166D mutant by the methods described by Christensen, Martin & Waley {(1990) Biochem . J . 266, 853-861} . Both the rate constants for acylation and deacylation for the hydrolysis of benzylpenicillin were decreased about 2000-fold in this mutant . In the K73R mutant, and in the double mutant, the rate constants for acylation were decreased about 100-fold and 10,000-fold respectively . All three mutants also had lowered values for the rate constants for the formation and dissociation of the non-covalent enzyme-substrate complex . The specificities of the mutants did not differ greatly from those of wild-type beta-lactamase, but the hydrolysis of cephalosporin C by the K73R mutant gave 'burst' kinetics.

Eur J Biochem, 1990 Dec 12, 194(2), 423 - 30
The ATPase of Bacillus alcalophilus . Purification and properties of the enzyme; Hoffmann A et al.; The ATPase of Bacillus alcalophilus was extracted from the bacterial membranes with Triton X-100 and purified by hydroxyapatite column chromatography . SDS gel-electrophoresis of the purified protein indicated the typical subunit pattern of an F1F0 structure with five F1 subunits (alpha, beta, gamma, delta, epsilon) and three F0 subunits (a,b,c) . The alpha and beta subunits were antigens for an antiserum against the corresponding subunits of the ATPase of Escherichia coli . Subunit c was extracted from the bacterial membranes with chloroform/methanol . Its amino acid composition was in the range of subunits c from other ATPases . Maximal ATPase activity was observed in the presence of 2-5 mM MgCl2, an ATP/Mg2+ ratio of 2:1 and 25% methanol . In the absence of methanol, only about 1% of the maximal activity was observed . The enzyme was also activated by Ca2+ (in the absence of methanol), reaching about 30% of the maximal activity . The dependence of initial velocity versus ATP of the Ca2(+)-activated but not of the Mg2+/methanol-activated enzyme indicted cooperativity with three strongly cooperative binding sites.

Biochemistry, 1990 Dec 11, 29(49), 10971 - 7
Folding and unfolding of the protoxin from Bacillus thuringiensis: evidence that the toxic moiety is present in an active conformation; Choma CT et al.; The action of trypsin or papain on the 130-kDa crystal protein (protoxin) from Bacillus thuringiensis subsp . kurstaki HD-73 yields a 67-kDa proteinase-resistant toxic fragment (toxin) which is derived from the N-terminal half of the molecule . Sensitivity to proteolysis and fluorescence emission spectroscopy showed that the toxin unfolded to a much greater extent in 6 M guanidinium chloride (GuHCl) than in 8 M urea . Protoxin also unfolded extensively in 6 M GuHCl, whereas in 8 M urea only the C-terminal half of the molecule had unfolded extensively . Both unfolded protoxin and unfolded toxin refolded to their native and biologically active conformations . The biphasic unfolding observed for protoxin suggests that the C-terminal half of the molecule unfolded rapidly, whereas the N-terminal toxic moiety unfolded at a much slower rate, similar to that of the free 67-kDa toxin . A 67-kDa fragment, derived from the N-terminal half of the molecule, could be generated from the protoxin in the presence of either urea or GuHCl by treatment with proteinases . Compared to toxin in denaturants, this fragment was found to be more sensitive to proteolysis . However, on removal of the denaturants the fragment had the same proteinase resistance and cytolytic activity as native toxin . The increased proteinase sensitivity of the fragment generated in the presence of denaturants appears to be due to a perturbation in the conformation of the N-terminal toxic moiety . This perturbation is attributed to the unfolding of the C-terminal region of the protoxin prior to its proteolysis to yield the 67-kDa fragment.(ABSTRACT TRUNCATED AT 250 WORDS)

FEBS Lett, 1990 Dec 10, 276(1-2), 14 - 6
Proteolysis of Bacillus stearothermophilus IF2 and specific protection by GTP; Severini M et al.; Translation initiation factor IF2 from Bacillus stearothermophilus (741 amino acids, Mr = 82,043) was subjected to trypsinolysis alone or in the presence of GTP . Following electroblotting and automated amino acid sequencing of the resulting peptides, the location and the sequential order of the main cleavage sites were identified . Trypsinolysis of IF2 ultimately generates two compact domains: a 24.5 kDa C-terminal fragment and a 40 kDa G-fragment which is obtained only in the presence of GTP which strongly protects a cleavage site within the GTP binding domain.

J Biol Chem, 1990 Dec 5, 265(34), 20923 - 30
Specificity-determining regions of a lepidopteran-specific insecticidal protein produced by Bacillus thuringiensis; Schnepf HE et al.; The lepidopteran-specific, insecticidal crystal proteins of Bacillus thuringiensis vary in toxicity to different species of lepidopteran larvae . We report studies of CryIA(a) and CryIA(c), two related proteins that have different degrees of toxicity to Heliothis virescens yet very similar degrees of toxicity to Manduca sexta . The amino acid differences between these proteins are located primarily between residues 280 and 722 . We have constructed a series of chimeric proteins and determined their toxicities to both insects . The most significant findings arise from the replacement of three segments of the cryIA(c) gene with homologous portions of the cryIA(a) gene: codons 332-428, 429-447, and 448-722 . Each of these segments contributed substantially and largely additively toward efficacy for H . virescens . However, replacement of the 429-447 segment of cryIA(c) gene with the cryIA(a) sequence resulted in a 27-50-fold reduction in toxicity toward M . sexta whereas the reduction in toxicity to H . virescens was only 3-4-fold . Subdivision of the 429-447 segment and replacements involving residues within this segment reduced toxicity to M . sexta by 5- to more than 2000-fold whereas toxicity to H . virescens was only reduced 3-10-fold . These observations indicate that: 1) different but overlapping regions of the cryIA(c) gene determine specificity to each of the two test insects; 2) some of the examined gene segments interact in determining specificity; and 3) different sequences in the cryIA(a) and cryIA(c) genes are required for maximal toxicity to M . sexta.

J Tenn Med Assoc, 1990 Dec, 83(12), 603 - 4
Infective endocarditis due to the CDC group M6 bacillus; Rose RC 3rd et al.; Although staphylococcal endocarditis has occasionally been associated with a TTP-like syndrome, a similar syndrome has not been reported with endocarditis due to aerobic gram-negative rods . We report a case of subacute bacterial endocarditis with a thrombocytopenic syndrome that at first resembled TTP which was due to an unusual gram-negative rod . This case emphasizes the need for repeated examination of the bacteremic patient to detect the changing murmurs of endocarditis.

Int J Food Microbiol, 1990 Dec, 11(3-4), 337 - 44
Toxicological, nutritional and microbiological evaluation of tempe fermentation with Rhizopus oligosporus of bitter and sweet apricot seeds; Tuncel G et al.; Bitter and sweet apricot seeds are by-products of the apricot processing industry . Bitter seeds, in particular, contain toxic levels of the cyanogenic substance amygdalin . Tempe was made from both kinds of seeds . The bitter seeds contain antimicrobial substances which must be removed by leaching and boiling prior to tempe fermentation . Apricot seed tempe had an agreeable taste . It contained approx . 21% (w/w) crude protein, 52% (w/w) crude fat, 1.5% (w/w) crude fibre and 25.5% (w/w) carbohydrates based on dry matter . The extent of biological acidification during soaking prior to fungal inoculation was inadequate to prevent growth of Bacillus cereus, and requires further optimisation . Bitter seeds were detoxified by the tempe process (approx . 70% of total cyanide was removed) . However, additional improvement of the detoxification process is required to obtain a completely safe product.

Pediatr Infect Dis J, 1990 Dec, 9(12), 890 - 3
Outbreak of Bacillus Calmette-Guérin-associated lymphadenitis and abscesses in Jamaican children; Praveen KN et al.; An outbreak of axillary lymphadenitis and abscesses after Bacillus Calmette-Guerin (BCG) vaccination (Pasteur Paris, Batch N5122) occurred in 139 Jamaican children between January and July, 1988 . The overall rate of this complication was 0.95% . The attack rate was 1.92% among the 0- to 6-week age group and 0.6% in the 7- to 52-week age group . Of 139 patients there were 77 males and 62 females and the mean age at presentation was 4 months . The mean size of the BCG scar, duration of healing and Mantoux reaction size differed significantly in patients compared with those of control infants (P less than 0.01) . Mycobacterium bovis was isolated from 11 patients . Immunologically index patients and controls differed significantly only with respect to T lymphocyte subpopulation percentages and concanavalin A stimulation indices . Evaluation of the BCG vaccine did not reveal either increased potency or microbial contamination . We conclude that increased susceptibility to the Pasteur strain of BCG might have contributed to the increased incidence of complications in these Jamaican children and hence caution should be exercised in switching one vaccine for another as is often done in the developing countries.

Can J Cardiol, 1990 Dec, 6(10), 461 - 2
Cardiobacterium hominis endocarditis; DeGuise M et al.; A 53-year-old man presented with nonspecific symptoms, evidence of aortic valve regurgitation and hepatosplenomegaly . Blood cultures grew Cardiobacterium hominis after 14 days of incubation . Endocarditis is caused by fastidious organisms such as C hominis in less than 5% of cases . To date approximately 40 cases of endocarditis due to this pleomorphic facultative anaerobic Gram-negative bacillus have been reported . C hominis is part of the normal human mouth flora.

Eur J Immunol, 1990 Dec, 20(12), 2651 - 9
Mycobacterial-induced cytotoxic T cells as well as nonspecific killer cells derived from healthy individuals and leprosy patients; Kaleab B et al.; Little information is available about the generation and specificity of the cytotoxic cells that eliminate human monocytes/macrophages infected with mycobacteria . To address this we have developed a cytotoxicity assay in which 51Cr-labeled monocytes pulsed with bacillus Calmette Guerin (BCG) or Mycobacterium leprae, were used as target cells in overnight cytotoxicity assays . As effector cells, peripheral blood mononuclear cells from healthy occupational contacts or from leprosy patients stimulated with antigen for 7 days were used . Cytotoxicity against antigen-pulsed monocytes that could be induced by mycobacterial antigens was proportional to the degree of antigen responsiveness in each individual, as measured in lymphocyte transformation tests . The lepromatous leprosy patients tested were often poor responders to BCG as well as M . leprae, both with regard to induction of cytotoxicity as well as in lympho-proliferation . Killing was significantly higher against antigen-pulsed vs . nonpulsed monocytes, although significant killing was induced against the latter as well and paralleled by induction of natural killer activity against the K-562 target cell . Cross-reactivity was observed between BCG and M . leprae, but not with unrelated antigen (tetanus toxoid) or with endogenous stress proteins induced by heat shock . M . leprae- and BCG-activated cytotoxic cells were found in both the CD4-CD8+ and CD4+CD8- populations, whereas in contrast the soluble antigen, purified protein derivative of M . tuberculosis, generated cytotoxic cells that were exclusively of the CD4+ phenotype . The involvement of both specific T cells as well as nonspecific cells in the killing of human macrophages may be important with respect to protection and immunopathology induced by mycobacterial antigens.

Gynecol Oncol, 1990 Dec, 39(3), 239 - 43
A randomized trial of cyclophosphamide, doxorubicin, and cisplatin with or without bacillus Calmette-Guerin in patients with suboptimal stage III and IV ovarian cancer: a Gynecologic Oncology Group study; Creasman WT et al.; Four hundred and eleven evaluable patients with suboptimal (greater than 1 cm residual) stage III and IV and recurrent ovarian cancer after surgical exploration and tumor debulking were prospectively randomized to receive cyclophosphamide, doxorubicin, and cisplatin (CAP) with or without bacillus Calmette-Guerin (BCG) . Therapy was planned for eight courses with the BCG to be given by the sacrification technique on Days 8 and 15 of each course . The addition of BCG did not improve response rate, progression-free interval (PFI), or survival . In a multivariate analysis prognostic factors significantly favorable for survival include nonmeasurable disease and young age . Those patients having tumor with a mucinous histology had poorer survival and PFI than patients with tumors composed of other cell types.

Blood, 1990 Dec 1, 76(11), 2229 - 34
Species specificity of human interleukin-3 demonstrated by cloning and expression of the homologous rhesus monkey (Macaca mulatta) gene; Burger H et al.; To enable preclinical studies on homologous interleukin-3 (IL-3) in primate species, we isolated the gene encoding Rhesus monkey IL-3 (RhIL-3) . The nucleotide sequence of the RhIL-3 gene displayed 92.9% homology with that of the human IL-3 (hIL-3) gene . The isolated RhIL-3 gene encodes a 143-amino acid (aa) precursor polypeptide, nine C-terminal residues shorter than the human protein . Protein homology was found to be 89.5% for the signal peptide (19 aa) and 80.5% for the mature protein (124 aa) . Comparison of the human and RhIL-3 coding sequences showed that the majority of substitutions had occurred at amino acid replacement sites indicating a rapid evolution of the IL-3 protein . After expression of a genomic fragment in COS cells, RhIL-3 cDNA was constructed, which enabled large-scale production of the RhIL-3 polypeptide, RhIL-3 produced by Bacillus licheniformis and purified to homogeneity appeared to be approximately 100-fold more effective in stimulating Rhesus monkey hematopoietic progenitors than hIL-3, whereas RhIL-3 and hIL-3 showed comparable stimulatory activity on normal as well as malignant human hematopoietic cells . Thus, the rapid evolution of hIL-3 has resulted in a unidirectional species specificity, which most likely restricts the in vivo effects of hIL-3 in Macaca species.

J Bacteriol, 1990 Dec, 172(12), 7301 - 5
Alveolysin, the thiol-activated toxin of Bacillus alvei, is homologous to listeriolysin O, perfringolysin O, pneumolysin, and streptolysin O and contains a single cysteine; Geoffroy C et al.; The gene coding for alveolysin, the thiol-activated toxin produced by Bacillus alvei, has been cloned by means of an oligonucleotide based on the known N-terminal sequence of the secreted protein . The complete nucleotide sequence of the gene has been determined . The deduced amino acid sequence of alveolysin shows that alveolysin shares homologies with listeriolysin O, perfringolysin O, pneumolysin, and streptolysin O . Alveolysin, like the other members of the family, contains a single cysteine in the conserved peptide sequence ECTGLA WEWWR.

J Bacteriol, 1990 Dec, 172(12), 7282 - 3
DNA sequences required for the alkalophily of Bacillus sp . strain C-125 are located close together on its chromosomal DNA; Kudo T et al.; Two alkali-sensitive mutants of alkalophilic Bacillus sp . strain C-125 were obtained . Mutant 38154 showed defective regulation of internal pH . Plasmids pALK1 and pALK2, containing DNA fragments different from those of the parent strain, were able to recover alkalophily in mutants 18224 and 38154, respectively . DNA analysis suggested that the two fragments overlapped on the chromosomal DNA of strain C-125.

J Bacteriol, 1990 Dec, 172(12), 6783 - 8
A novel Bacillus thuringiensis gene encoding a Spodoptera exigua-specific crystal protein; Visser B et al.; Only one of the four lepidoptera-specific crystal protein subclasses (CryIC) Bacillus thuringiensis was previously shown to be highly toxic against several Spodoptera species . By using a cryIC-derived nucleotide probe, DNA from 25 different strains of B . thuringiensis was screened for the presence of homologous sequences . A putative crystal protein gene, considerably different from the cryIC gene subclass, was identified in the DNA of strain 4F1 (serotype kenyae) and cloned in Escherichia coli . Its nucleotide sequence was determined and appeared to contain several features typical for a crystal protein gene . Furthermore, the region coding for the N-terminal part of the putative toxic fragment showed extensive homology to subclass cryIA sequences derived from gene BtII, whereas the region coding for the C-terminal part appeared to be highly homologous to the cryIC gene BtVI . With an anti-crystal protein antiserum, a polypeptide of the expected size could be demonstrated in Western immunoblots, onto which a lysate of E . coli cells harboring the putative gene, now designated as BtXI, had been transferred . Cells expressing the gene appeared to be equally toxic against larvae of Spodoptera exigua as recombinant cells expressing the BtVI (cryIC)-encoded crystal protein . However, no toxicity against larvae of Heliothis virescens, Mamestra brassicae, or Pieris brassicae could be demonstrated . The nucleotide sequence analysis and the toxicity studies showed that this novel crystal protein gene falls into a new cryl gene subclass . We propose that this subclass be referred to as cryIE.

J Bacteriol, 1990 Dec, 172(12), 6759 - 63
Deletion analysis of the 51-kilodalton protein of the Bacillus sphaericus 2362 binary mosquitocidal toxin: construction of derivatives equivalent to the larva-processed toxin; Clark MA et al.; Bacillus sphaericus 2362 produces a binary toxin consisting of 51- and 42-kDa proteins, both of which are required for toxicity to mosquito larvae . Upon ingestion by larvae, these proteins are processed to 43 and 39 kDa, respectively . Using site-directed mutagenesis, we have obtained N- and C-terminal deletions of the 51-kDa protein and expressed them in B . subtilis by using the subtilisin promoter . Removal of 21 amino acids from the N terminus and 53 amino acids from the C terminus resulted in a protein with the same electrophoretic properties as the 43-kDa degradation product which accumulates in the guts of mosquito larvae . This protein was toxic only in the presence of the 42-kDa protein . A deletion of 32 amino acids at the N terminus combined with a 53-amino-acid deletion at the C terminus resulted in a protein which retained toxicity . Toxicity was lost upon a further deletion of amino acids at potential chymotrypsin sites (41 at the N terminus, 61 at the C terminus) . Comparison of the processing of the 51- and the 42-kDa proteins indicated that in spite of their sequence similarity proteolysis occurred at different sites.

Am J Respir Cell Mol Biol, 1990 Dec, 3(6), 563 - 70
Fibronectin is not detectable on the intact buccal epithelial surface of normal rats or humans; Mason CM et al.; Fibronectin (FN) has been postulated to prevent gram-negative bacillary (GNB) colonization of the oropharynx by covering epithelial cell GNB receptors . We investigated the distribution of FN along the luminal surface of oropharyngeal epithelium in animals and humans . Examination of buccal epithelial biopsies obtained from normal rats revealed no luminal surface FN by either immunofluorescent or immunoperoxidase staining . Extraction of epithelial surface proteins and quantitation of FN by rocket immunoelectrophoresis and electrophoretic transfer to nitrocellulose followed by immunologic detection also detected no FN from normal animals' oropharyngeal biopsies . Buccal epithelial biopsies from three normal humans were examined for FN using electrophoretic transfer to nitrocellulose followed by immunologic detection, and no FN was demonstrable . Our results suggest that FN is not present on the oral epithelial surface of healthy rodents or humans, and that FN may not be involved in the pathogenesis of bacillary colonization.

Arch Neurol, 1990 Dec, 47(12), 1313 - 7
Tuberculous meningitis . Short course of chemotherapy; Alarcon F et al.; In March 1986, we began a 6-month short course trial of therapy for tuberculous meningitis, in which 28 patients were analyzed . The diagnosis was based on the following cerebrospinal fluid test results: in 53.5% of the cases, Mycobacterium tuberculosis was identified by direct smear; in 57%, culture in Lowenstein-Jensen medium was positive; in 83.3%, the detection of anti-bacille Calmette-Guerin (BCG) antibodies by enzyme-linked immunosorbent assay was positive; and in 74%, the dosification of adenosine deaminase activity was positive . In addition, in 21.4% of the cases, the diagnosis was established by means of autopsy findings . Moreover, the diagnosis was supported by bacteriological analyses from another tissue or body fluids . Despite the administration of an antituberculous therapy, 32.4% of the patients died: all of the decreased had reached the last stage of the disease by the beginning of treatment . Sixteen percent of the patients who survived after more than 18 months of follow-up after therapy had ended suffered neurological sequelae . With the 6-month therapeutic regimen, the morbidity/mortality is similar to that found in the longer-course therapies . The latter regimen is therefore thought to be a good and acceptable therapeutic option for the treatment of tuberculous meningitis.

J Urol, 1990 Dec, 144(6), 1399 - 400
Clinical and pathological findings in prostates following intravesical bacillus Calmette-Guerin instillations; Mukamel E et al.; The prostates of 36 patients who were treated with intravesical bacillus Calmette-Guerin were evaluated by digital rectal examination and transrectal ultrasonography . When abnormal palpatory and/or ultrasonographic findings were detected, core needle biopsies from the suspicious areas were performed . Of the 36 patients 20 underwent biopsies of the prostate . Pathological findings revealed typical granulomas in 8 patients (3 caseating and 5 noncaseating multifocal granulomas) . Nonspecific chronic prostatitis was noted in 4 patients and benign prostatic hyperplasia was noted in 8 . The number of bacillus Calmette-Guerin instillations ranged from 6 to 19 . The interval from initiation of therapy to biopsy ranged from 1.5 to 14.5 months . Caseating granulomas were found during the early course of bacillus Calmette-Guerin instillations (1.5 to 3.0 months), whereas noncaseating granulomas were detected at later stages (4 to 14.5 months) . These findings present a high incidence of granuloma formation in patients treated with intravesical bacillus Calmette-Guerin . The duration of therapy is a determinant factor in the induction of granuloma type.

J Urol, 1990 Dec, 144(6), 1362 - 4
Adverse impact of fibrin clot inhibitors on intravesical bacillus Calmette-Guerin therapy for superficial bladder tumors; Hudson MA et al.; Although intravesical bacillus Calmette-Guerin therapy has proved to be efficacious in the treatment and prophylaxis against tumor recurrence of superficial bladder tumors, its mechanism of action has not been fully elucidated . Previous work has suggested that bacillus Calmette-Guerin organisms attach to the matrix protein, fibronectin, during fibrin clot formation at sites of urothelial disruption and that this attachment was required for the antitumor effect of bacillus Calmette-Guerin to be expressed . Furthermore, drugs inhibiting clot formation were found to abrogate the antitumor effect of intravesical bacillus Calmette-Guerin therapy in a murine bladder tumor model . To examine the effect of inhibitors of fibrin clot formation on the results of intravesical bacillus Calmette-Guerin therapy, a retrospective analysis of 149 evaluable patients receiving intravesical bacillus Calmette-Guerin for superficial bladder tumors was performed . The over-all response rate free of tumor for 29 patients who concomitantly received inhibitors of fibrin clot formation with bacillus Calmette-Guerin therapy was 48%, as compared with 67% for 120 patients who were not receiving these medications (p = 0.0655, chi-square) . The most striking difference was noted for patients who failed with recurrent superficial disease . Of the patients who received fibrin clot inhibitors during intravesical bacillus Calmette-Guerin therapy 35% had recurrent superficial tumors compared to only 8% of those who did not receive these drugs during a mean followup of 29.8 plus or minus 11 months (p = 0.005, chi-square) . Our study suggests that inhibitors of fibrin clot formation may have an adverse influence on the results of intravesical bacillus Calmette-Guerin therapy for superficial bladder tumors.

J Urol, 1990 Dec, 144(6), 1331 - 3; discussion 1333-4
Fatal disseminated mycobacterial infection following intravesical bacillus Calmette-Guerin; Deresiewicz RL et al.; We describe a fatal case of disseminated mycobacteriosis after intravesical bacillus Calmette-Guerin immunotherapy . We summarize the prior safety record of this therapeutic modality, discuss local and systemic pathophysiological mechanisms by which dissemination might have occurred, and review the reported clinical experience with antituberculous chemotherapy for significant bacillus Calmette-Guerin infection . Finally, we offer suggestions for prophylaxis of certain patients with a history of exposure to intravesical bacillus Calmette-Guerin.

J Urol, 1990 Dec, 144(6), 1328 - 30
Fatal sepsis following intravesical bacillus Calmette-Guerin administration for bladder cancer; Rawls WH et al.; Intravesical administration of bacillus Calmette-Guerin has been shown to be highly effective treatment of superficial bladder cancer . Complications from bacillus Calmette-Guerin therapy are usually minor but serious and even fatal reactions can occur . Five recent cases illustrate the gravity of bacillus Calmette-Guerin sepsis . One man with severe debility and the organic brain syndrome died acutely with a fever of 40 C . Two men had frank sepsis that progressed to multiorgan failure and death . Sepsis progressed despite the use of isoniazid, rifampin and streptomycin . Two men who had equally progressive sepsis with intravesical bacillus Calmette-Guerin survived with the use of cycloserine for the first 72 hours of treatment . Triple antituberculous antibiotics, including cycloserine, may be lifesaving . Sepsis resulted from intravenous absorption through inflamed or disrupted urothelium . Bacillus Calmette-Guerin treatment should not be administered in the presence of severe cystitis or after grossly traumatic catheterization.

Gastroenterology, 1990 Dec, 99(6), 1723 - 6
Host response to mycobacterial infection in the alcoholic rat; Mendenhall CL et al.; Animals, chronically treated with alcohol, were inoculated with mycobacteria (bacillus Calmette-Guerin, 10.2 x 10(6) organisms) into the spleen to produce a granulomatous hepatitis . Before infection, chronic alcohol ingestion was associated with a depressed skin test response to phytohemagglutinin, 71.7% of baseline (P = 0.009) . Mycobacterial (bacillus Calmette-Guerin) infection stimulated phytohemagglutinin skin test response to 417% of baseline in controls and 299% in alcoholics (P less than 0.001) . The hepatic granuloma response was altered with smaller but more numerous granulomas (mean +/- SEM, 81.2 +/- 1.5 microns2 of area with a frequency of 1.8 granulomas per field in alcoholics vs . 129.8 +/- 5.71 microns2 and 1.2 granulomas per field in controls; P less than 0.001) . These changes were associated with a 10-fold increase in colony-forming units per gram of liver (54.5 +/- 18.2 in alcoholics vs . 5.6 +/- 1.83 in controls; P = 0.0006) . This model offers precise parameters for host response to infection and indicates that alcohol significantly impairs the clearing capacity for mycobacteria from the liver.

Arch Biochem Biophys, 1990 Dec, 283(2), 266 - 70
Resonance Raman investigation of a soluble cytochrome c552 from alkaliphilic Bacillus firmus RAB; Larsen RW et al.; The environment of the heme site of a low-potential soluble cytochrome (c552) from alkaliphilic Bacillus firmus RAB has been characterized with resonance Raman scattering and compared to that of horse heart cytochrome c . The Raman data indicate that vibrational bands sensitive to the axial ligation of the heme, as well as modes sensitive to the heme peripheral environment in cytochrome c552, are distinct from those of horse heart cytochrome c . The spectra of cytochrome c552 display resonance Raman modes indicative of a methionine as the sixth ligand in the oxidized form, while the reduced form appears to contain a nitrogenous-based sixth ligand . In addition, Q-band excitation reveals differences among vibrational modes in cytochrome c552 that are sensitive to the amino acid environment surrounding the heme.

J Bacteriol, 1990 Dec, 172(12), 6689 - 96
Structural and genetic organization of IS232, a new insertion sequence of Bacillus thuringiensis; Menou G et al.; In the Bacillus thuringiensis strains toxic for the lepidopteran larvae, the delta-endotoxin genes cryIA are frequently found within a composite transposonlike structure flanked by two inverted repeat sequences . We report that these elements are true insertion sequences and designate them IS232 . IS232 is a 2,184-bp element and is delimited by two imperfect inverted repeats (28 of 37 bp are identical) . Two adjacent open reading frames, overlapping for three codons, span almost the entire sequence of IS232 . The potential encoded polypeptides of 50 and 30-kDa are homologous to the IstA and IstB proteins of the gram-negative insertion sequence IS21 . The N-terminal part of the 50-kDa polypeptide contains a helix-turn-helix DNA-binding motif . The junctions at the insertion sites of three IS232 elements were analyzed . Each case was different, with 0, 4, or 6 bp of the target DNA being duplicated . Transposition of IS232 in Escherichia coli was demonstrated by using a genetic marker inserted upstream of the two open reading frames.

Zhonghua Jie He He Hu Xi Za Zhi, 1990 Dec, 13(6), 333 - 5, 379
{Drug resistant plasmids of Pseudomonas pyocyanea in the respiratory tract}; Xu W; This paper reports the results of the conjugating transmission test, the plasmid elimination test and the plasmid analysis of pseudomonas pyocyanea which were separated from the sputum of the patients with infection of respiratory tract . The result of the conjugating test and the plasmid elimination test of P6P8 strains indicated that the partial drug resistances of two strains are mediated by the drug resistant plasmids . The results of the plasmids analysis in 14 strains of pseudomonas pyocyanea showed that this pyocyanic infection was mainly caused by the bacillus pyocyanea carrying 38 x 10(6) dal plasmids . The technique of plasmid analysis is simple and good repetition . The specificity of this method is analogous to phage typing . This method is worth popular in the epidemic investigation of pathogenic bacteria tracing.

J Gen Microbiol, 1990 Dec, 136 ( Pt 12), 2377 - 83
Cloning of two genes from Bacillus circulans WL-12 which encode 1,3-beta-glucanase activity; Fiske MJ et al.; Two genes encoding distinct 1,3-beta-glucanases have been cloned from Bacillus circulans and expressed in Escherichia coli . A cosmid library of B . circulans WL-12 DNA was constructed in the broad-host-range cosmid pLAFR1 and screened in E . coli for clones which exhibited 1,3-beta-glucanase activity . Two 1,3-beta-glucanase-positive clones were identified which contained genes encoding two independent 1,3-beta-glucanases as shown by biochemical, physical and molecular analyses . The cosmids, designated pMON5401 (27.1 kb insert) and pMON5402 (24.7 kb insert), encoded 68 kDa and 40 kDa 1,3-beta-glucanases, respectively . Both 1,3-beta-glucanases were purified from their respective E . coli strains, characterized biochemically, and were shown to exhibit lytic activity against purified yeast cell wall preparations.

EMBO J, 1990 Dec, 9(12), 3907 - 12
Evidence for a role of phosphatidylcholine-hydrolysing phospholipase C in the regulation of protein kinase C by ras and src oncogenes; Diaz-Laviada I et al.; The products of ras and src oncogenes are thought to be important components in pathways regulating cell proliferation and differentiation . In fibroblasts transformed by these oncogenes, increased diacylglycerol levels have been found which most probably arise from activation of the turnover of phosphatidylcholine . Diacylglycerol is a key activator of protein kinase C whose role in cell growth and transformation has been proposed . We demonstrate here by using immunochemical techniques that transformation by ras or src oncogenes is associated with permanent translocation of protein kinase C to the cytoplasmic membrane . However, no down-regulation of the enzyme is observed despite its permanent activation in these transformants . Importantly, the lack of down-regulation observed in ras and src transformed cell lines is mimicked by chronic treatment of NIH 3T3 fibroblasts with exogenous Bacillus cereus phosphatidylcholine-hydrolysing phospholipase C, but not with phorbol myristate acetate or exogenous Bacillus thuringiensis phosphatidylinositol-hydrolysing phospholipase C . These results strongly suggest that diacylglycerol derived from phosphatidylcholine but not from phosphoinositide turnover is responsible for the atypical regulation of protein kinase C in cell lines transformed by ras and src oncogenes.

J Am Mosq Control Assoc, 1990 Dec, 6(4), 667 - 71
Integrated use of planaria (Dugesia dorotocephala) and Bacillus thuringiensis var . israelensis against Aedes taeniorhynchus: a laboratory bioassay; Perich MJ et al.; The effectiveness of integrating Bacillus thuringiensis var . israelensis (B.t.i.) and the predatory planaria, Dugesia dorotocephala against Aedes taeniorhynchus was determined under controlled laboratory conditions . There was no significant effect (P greater than 0.05) of B.t.i . on D . dorotocephala either by direct association or through ingestion of B.t.i . dosed larvae . Planaria, alone, and B.t.i . combined with planaria, both provided significant (P less than 0.05) reduction of Ae . taeniorhynchus populations through the 12-week evaluation.

Biomed Environ Sci, 1990 Dec, 3(4), 397 - 405
Use of in situ structural and functional variables of phytoplankton of the river Ganga for assessment of heavy metal toxicity; Singh AK et al.; Toxicity of Cd and Zn on autotrophic index, pigment diversity . 14C uptake, and in situ nitrogenase activity of phytoplankton of the river Ganga has been studied for the first time in India using CEPEX enclosures . Maximum reduction in algal population was noted at 8.0 micrograms ml-1 Cd followed by 8.0 micrograms ml-1 Zn . Complete elimination of two and four species was observed respectively at 4.0 and 8.0 micrograms ml-1 Cd and Zn . The filamentous forms showed greater tolerance against Cd and Zn, whereas unicellular forms were more sensitive to test metals used . Bacillariophytes in general depicted greater sensitivity for both the metals . A concentration-dependent metal-specific increase in autotrophic index and pigment diversity of phytoplankton was noted for Cd and Zn . Inhibition of carbon and nitrogen fixation was, however, concentration dependent and metal specific . Looking at the sensitivity of 14CO2 uptake rather than other variables, we recommend the employment of this parameter for assessment of heavy metal toxicity in an aquatic ecosystem.

Mol Microbiol, 1990 Dec, 4(12), 2137 - 46
Molecular characterization of immune inhibitor A, a secreted virulence protease from Bacillus thuringiensis; Lovgren A et al.; The gene for the secreted neutral metalloprotease, immune inhibitor A (InA), from Bacillus thuringiensis var . alesti has been cloned and sequenced . The deduced amino acid sequence has been confirmed by partial amino acid sequencing . The central part of the amino acid sequence showed similarity to the active site in thermolysin . Southern and Western blots show that InA-related sequences are common among other B . thuringiensis subspecies . In Western blots, 17 out of 25 tested species gave a positive signal . Culture filtrates from subspecies expressing InA were toxic when injected in Trichoplusia ni larvae, whereas filtrate from a strain negative in Western blot had no effect when injected . The LD50 dose of purified InA protein injected in T . ni larvae was 12.5 +/- 2.5 ng per mg of larval body weight.

Mol Microbiol, 1990 Dec, 4(12), 2087 - 94
Activation of a cryptic crystal protein gene of Bacillus thuringiensis subspecies kurstaki by gene fusion and determination of the crystal protein insecticidal specificity; Dankocsik C et al.; DNA hybridization with the insecticidal crystal protein gene cryllA (formerly cryBl) of Bacillus thuringiensis supspecies kurstaki has shown that subspecies kurstaki contains a cryllA-related sequence in addition to the cryllA gene (Donovan et al., 1988a) . We have cloned the cryllA-related sequence and have determined that the sequence, which has been designated cryllB, is 89% identical to the cryllA gene . Recombinant B . thuringiensis cells harbouring the cloned cryllB gene produced very little CryllB protein . A high level of production of the CryllB protein was achieved by fusing the regulatory region of the crylllA crystal protein gene to the cryllB gene . The CryllB protein was found to be highly toxic to Lymantria dispar, Heliothis virescens and Trichoplusia ni, and was not toxic to Aedes aegypti.

Antimicrob Agents Chemother, 1990 Dec, 34(12), 2426 - 8
Activity of cefixime against Helicobacter pylori and affinities for the penicillin-binding proteins; Ikeda F et al.; Cefixime induced the formation of rounded cells from the spiral bacillary form of Helicobacter pylori at the MIC or less . Three main penicillin-binding proteins, called A, B and C, were separated from H . pylori . Cefixime had the strongest affinity to penicillin-binding protein B . The binding of cefixime to this protein may induce the formation of rounded H . pylori cells.

Can J Microbiol, 1990 Dec, 36(12), 870 - 8
Interaction of the Bacillus sphaericus mosquito larvicidal proteins; Davidson EW et al.; Genes for 51.4- and 41.9-kDa insecticidal proteins of Bacillus sphaericus were separately cloned and expressed in Escherichia coli . Both proteins were required for toxicity . Approximately equal numbers of cells containing the 51.4- and 41.9-kDa proteins produced the greatest toxicity; excess 41.9-kDa protein did not affect toxicity, whereas excess 51.4-kDa protein reduced activity . Larvae were killed when 41.9-kDa protein was fed up to 24 h after the 51.4-kDa protein, but not when the order of feeding was reversed . Radiolabelled toxins bound in approximately equal amounts to the gastric caecum and posterior midgut of Culex quinquefasciatus larvae . Radiolabelled 51.4-kDa protein was rapidly degraded by ca . 12-13 kDa in the larval gut, while 41.9-kDa protein was degraded by 1-2 kDa . Nonreduced toxin extracted from B . sphaericus produced a band on SDS-PAGE of ca . 68-74 kDa that contained both 51.4- and 41.9-kDa proteins based on sequence analysis, and a band of ca . 51 kDa that contained primarily 41.9-kDa protein . Escherichia coli containing 51.4-kDa protein enhanced toxicity of the latter eluted SDS-PAGE band . These proteins may associate very strongly, and trace amounts of 51.4-kDa protein in preparations of 41.9-kDa protein from B . sphaericus may be responsible for the previously reported toxicity of the latter.

Curr Genet, 1990 Dec, 18(6), 523 - 30
Gene disruption of the pcbAB gene encoding ACV synthetase in Cephalosporium acremonium; Hoskins JA et al.; Plasmid pPS96 was used to disrupt the genomic region immediately upstream of pcbC in C . acremonium by homologous integration . Approximately 4% of the C . acremonium transformants obtained with pPS96 were unable to produce beta-lactam antibiotics . All transformants obtained with other plasmids and isolates which had not been exposed to transforming DNA retained the ability to produce beta-lactams . Enzyme analysis showed that ACV synthetase activity was missing in the beta-lactam-minus pPS96 transformants . Southern copies of pPS96 in all beta-lactam-minus transformants analyzed . However, predictable alterations of the targeted region were not detected . Transformation of antibiotic-minus transformants with plasmid pZAZ4, carrying a wild-type copy of the region targeted for disruption, resulted in restoration of the ability to produce beta-lactams in greater than 80% of the transformants recovered . Location of the pcbAB gene upstream from pcbC was confirmed by comparing the amino acid sequence of internal peptides from purified ACV synthetase with that deduced from the DNA sequence of the region targeted for disruption . The direction of transcription of the pcbAB gene is opposite that of the pcbC gene . Further analysis of amino acid sequence data from ACV synthetase revealed regions of strong similarity with the peptide synthetases responsible for production of tyrocidine and gramicidin S in Bacillus brevis.

J Am Mosq Control Assoc, 1990 Dec, 6(4), 631 - 4
Efficacy of Bacillus sphaericus formulations against Psorophora columbiae larvae in small rice plots; Bowles DE et al.; Bacillus sphaericus formulations were evaluated against Psorophora columbiae larvae in small rice plots . All formulations tested provided good control (greater than 84%) for all rates tested at 2 days posttreatment . At 10 days posttreatment, fair control (greater than 70%) was recorded for ABG-6232 (0.184 kg/ha), ABG-6262; 27-242-BA (0.766 and 2.298 liters/ha), and ABG-6262; 29-293-BA (2.298 liters/ha) . At 7 days posttreatment mortality was below 66% . Control did not exceed 27% at 15 days posttreatment for any formulation tested . Application rates for all formulations tested had little relationship with observed mortality, and control levels often were greater at the lower dosages tested . Bacillus sphaericus provided an effective means for controlling Ps . columbiae, but additional research is needed to determine the lower limit of effective treatment dosages and spore persistence in the rice field environment.

Biochem Int, 1990 Dec, 22(5), 867 - 76
Phosphatidylinositol-attached 5'-nucleotidase from synaptic plasma membrane of rat brain; Lai KM et al.; Synaptic plasma membranes (SPM) of rat brain contained a 5'-nucleotidase that was specifically released by Bacillus thuringiensis phosphatidylinositol-specific phospholipase C (PIPLC) . About 30% of the enzyme was readily released and the remainder was less susceptible . Purified 5'-nucleotidase was treated with PIPLC and the resultant enzyme was almost totally partitioned into the detergent-poor phase following phase-separation in Triton X-114 indicating that PIPLC converted the enzyme from an amphipathic to a hydrophilic form . The results suggest that 5'-nucleotidase is anchored into SPM by a covalently attached phosphatidylinositol moiety.

Chem Phys Lipids, 1990 Dec, 56(2-3), 159 - 67
Preparation and application of an affinity matrix for phosphatidylinositol-specific phospholipase C; Shashidhar MS et al.; A non-hydrolyzable phosphonate analogue of phosphatidyl inositol, racemic myo-inosityl-(1)-5-oxa-16-trifluoroacetamidohexadecyl phosphonate, was synthesized . This phosphonate inhibited the activity of phosphatidyl inositol-specific phospholipase C (PI-PLC) from Bacillus cereus with an IC50 of approximately 10 mM . Removal of the trifluoroacetyl blocking group followed by covalent binding of the phosphonate to cyanogen bromide activated Sepharose 4B via the amino group produced an affinity matrix specific for the PI-PLC from B . cereus . This affinity matrix was used to purify the phospholipase C from a complex mixture of proteins in a single step . Competition experiments with myo-inositol in the elution medium indicated that specific binding of the enzyme to the matrix most likely involves the enzyme active site . The inositol phosphonate derivatized matrix was stable over several months in neutral and alkaline media and was used repeatedly without loss of binding capacity . These results show that affinity matrices employing myo-inositol phosphonate ligands are useful for isolation and binding studies of PI-PLC and possibly of other enzymes interacting with phosphoinositides or myo-inositol phosphate derivatives.

Mol Gen Mikrobiol Virusol, 1990 Dec, (12), 17 - 8
{Detection of aquatic microorganisms from the Black Sea--producers of restriction endonucleases}; Dedkov VS et al.; 300 clones of microorganisms isolated at different stations and from different depths in the Black Sea were screened for restriction endonucleases production . The production of restriction endonucleases was found in 17 clones screened . Three of them were identified to be Alteromonas haloplanktis B1 . Restriction endonuclease AhaB1 is an isoshizomer of Sau961 . An identified Alteromonas haloplanktis clone B8 produces AhaB8I restriction endonuclease the prototype to which is KpnI . Of the clones isolated three are Moraxella species B4 producing MapB4I restriction endonuclease analogous to BanI, three are Bacillus species producing BspB2I and one is Micrococcus lylae 113 producing Mly1131 analogue of NarI, six Moraxella species B6 produce MspB6I . The isolated producer strains may be used for isolation of above mentioned restriction endonucleases.

J Invest Dermatol, 1990 Dec, 95(6 Suppl), 180S - 184S
Role of interferons in the therapy of melanoma; Kirkwood JM et al.; A range of potent immunoregulatory molecules termed cytokines has become available for the therapy of human melanoma . Among the cytokines, the interferons (IFN) have been examined in great depth for the therapy of melanoma . IFN are able to modulate host effector cell function, including the tumor cytolytic function of lymphocytes and monocytes . IFN also have the capacity to regulate the distribution of circulating immunoregulatory (T) lymphocytes and the expression of tumor cell surface antigens, as well as class I and II products of the major histocompatibility locus . These activities of the IFN have led to their early application for treatment of human melanoma . The empirical evidence that IFN alpha exerts clinically significant anti-tumor effects against melanoma is reviewed, and evolving status of adjuvant trials of IFN alpha and gamma is noted . New indirect host-mediated anti-tumor activities that may potentially be manifest by IFN have yet to be fully harnessed . The opportunity to obtain meaningful anti-tumor activity in advanced disease or adjuvant settings, at dose ranges below those which are toxic (conventional maximal tolerable), are at hand . The U.S . cooperative groups {Eastern Cooperative Oncology Group (ECOG), Cancer and Leukemia Group B (CALGB), and South West Oncology Group (SWOG)} are studying IFN gamma in pursuit of this goal in advanced and adjuvant settings for melanoma and other tumors . The determination of the clinical role of IFN as biologic response modifiers demands equal commitment to the clinical assessment of immunobiologic mechanisms and anti-tumor effects . The immunologic assessment of IFN and a number of other cytokines is a major focus of the Pittsburgh Cancer Institute . Regional delivery of cytokines such as interleukin-2 (IL-2) may be the most appropriate and least toxic approach, given their half-life . Regional therapy by the intralesional route has yielded enhanced activity for a range of biologics, including bacillus Calmette-Guerin (BCG), IL-2, and tumor necrosis factor (TNF) . Intralymphatic therapy with methanol extraction residue of BCG (MER-BCG) has been tested, and trials are now in progress with IL-2 to assess the optimal dosage by this route . It is likely that the optimal role of IFN and other cytokines will be found in combination with one another, and with different biologic modalities such as monoclonal antibodies and vaccines, to allow expansion and heightened activity of the desired effector cell populations in the host . Enhanced host toxicities, as well as anti-tumor effects, may require that special attention be devoted to optimal sequence of administration to enhance the therapeutic index.

Agric Biol Chem, 1990 Dec, 54(12), 3227 - 33
Nucleotide sequence of the gene coding for the BanIII DNA methyltransferase in Bacillus aneurinolyticus; Kawakami B et al.; The gene coding for the ATCGAT specific BanIII DNA methyltransferase (M-BanIII) of Bacillus aneurinolyticus was cloned and its nucleotides sequenced . The coding region was assigned on the nucleotide sequence on the basis of the N-terminal amino acid sequence and molecular weight of the enzyme . The M-BanIII gene coded for a protein of 580 amino acid residues (MW 66,344) . Comparison with other methylases indicated that the M-BanIII sequence contained a segment of tetra-amino acids, NPPY, characteristic of N6-adenine methylases . In addition, some homologous regions were found in the sequences of type II adenine methylases PaeR7I(CTCGAG), TaqI(TCGA) and PstI(CTGCAG), containing TCGA within the recognition sequences.

Appl Microbiol Biotechnol, 1990 Dec, 34(3), 297 - 302
Efficient production of human alpha-amylase by a Bacillus brevis mutant; Konishi H et al.; A cDNA for mature human salivary alpha-amylase was directly joined to a sequence encoding the signal peptide of the middle wall protein (MWP) gene of Bacillus brevis 47 . This hybrid gene was placed downstream from the multiple promoter region of the MWP gene on a low copy-number plasmid vector, pHW1 . B . brevis 47 carrying the plasmid produced 0.9 mg/l of active human alpha-amylase in the medium . A B . brevis 47 mutant obtained on mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine produced an increased amount of the alpha-amylase (6 mg/l) . When the fused gene was inserted into a high copy-number expression vector, pNU200, and then introduced into the mutant, a large amount (60 mg/l) of the alpha-amylase was produced in the medium . The alpha-amylase showed approximately the same specific activity and molecular weight as those of the natural enzyme . The mutant showed higher sensitivity to various antibiotics than the original strain, and altered cell wall and cytoplasmic membrane protein compositions . The results of reversion analysis suggested that a single mutation is responsible for the above phenotypes and hyper-productivity of human alpha-amylase.

Eur J Biochem, 1990 Nov 26, 194(1), 95 - 102
Cloning and sequence analysis of the genes encoding the dihydrolipoamide acetyltransferase and dihydrolipoamide dehydrogenase components of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus; Borges A et al.; A 2641-bp EcoRI fragment of DNA that encodes the C-terminal part of the dihydrolipoyl acetyltransferase (E2) component and the dihydrolipoamide dehydrogenase (E3) component of the pyruvate dehydrogenase complex of Bacillus stearothermophilus has been cloned in Escherichia coli . Its nucleotide sequence was determined . A 705-bp truncated open reading frame was located at the 5'end of the insert which, together with the 588-bp truncated open reading frame at the 3' end of another EcoRI fragment of B . stearothermophilus DNA previously cloned and sequenced {Hawkins, C . F., Borges, A . & Perham, R . N . (1990) Eur . J . Biochem . 191, 337-446}, was identified as the gene, pdhC, encoding the E2 polypeptide chain . Direct sequence analysis of the purified E2 chain confirmed that the two EcoRI fragments are adjoining in the B . stearothermophilus genome . The E3 gene, pdhD, begins just 4 bp downstream from the stop codon of the pdhC gene . The amino acid sequences deduced from the pdhC and pdhD genes correspond to proteins of 427 amino acids (E2, Mr 46,265) and 469 amino acids (E3, Mr 49,193), respectively . Both genes are preceded by potential ribosome-binding sites and the E3 gene is followed by a stemloop structure characteristic of rho-independent transcription terminators . The B . stearothermophilus E2 and E3 chains exhibit substantial sequence similarity with the corresponding subunits of other 2-oxo-acid dehydrogenase multienzyme complexes . The cloning and sequence analysis described here complete the description of the gene cluster (pdhA, B, C and D) which encodes the B . stearothermophilus pyruvate dehydrogenase multienzyme complex.

Eur J Biochem, 1990 Nov 26, 194(1), 75 - 80
Properties of purified squalene-hopene cyclase from Bacillus acidocaldarius; Ochs D et al.; The squalene-hopene cyclase from Bacillus acidocaldarius cytoplasmic membrane, was purified to homogeneity by solubilization with Triton X-100, chromatography on DEAE-cellulose, phenyl Sepharose and two gel-filtration columns . The enzyme monomer had a molecular mass of 75 kDa . The sequence of the first 23 amino acids was determined by Edman degradation . The enzyme activity was efficiently inhibited by n-alkyldimethylammonium halides with alkyl chain lengths between 12 and 18 C atoms . Inhibition was also observed with (5-hydroxycarvacryl)trimethylammonium chloride 1-piperidine carboxylate, dodecyldimethylamine N-oxide, azasqualene and farnesol . Competitive inhibition with dodecyltrimethylammonium bromide, (5-hydroxycarvacryl)trimethylammonium chloride 1-piperidine carboxylate and dodecyldimethylamine N-oxide was demonstrated by Lineweaver-Burk plots.

Eur J Biochem, 1990 Nov 26, 194(1), 161 - 5
Delineation of the minimal portion of the Bacillus sphaericus 1593M toxin required for the expression of larvicidal activity; Sebo P et al.; The two genes of Bacillus sphaericus 1953M coding for the 51.4-kDa and 41.9-kDa proteins are both required for the expression of the active larvicidal toxin in Escherichia coli . The minimal size of the active peptide of the 41.9-kDa toxin was defined by in vitro deletion analysis of the gene and found to consist of 338 amino acids (38.3 kDa) . N-terminal deletions past the Ile18 residue and C-terminal deletions past the His352 residue result in the loss of toxic activity and rapid degradation of such modified toxins by host proteases . The minimal active 38.3-kDa peptide produced in E . coli seems to mimick the stable processed form of the toxin found in larval midguts . However, it still requires the action of the synergistic 51.4-kDa protein for the larvicidal activity.

J Biol Chem, 1990 Nov 25, 265(33), 20547 - 54
Purification and reconstitution of the F1F0-ATP synthase from alkaliphilic Bacillus firmus OF4 . Evidence that the enzyme translocates H+ but not Na+; Hicks DB et al.; The F1F0-ATP synthase from the alkaliphilic Bacillus firmus OF4 was purified in a reconstitutively active form, in good yield and with a high specific ATPase activity when appropriately activated . The purification procedure involved octyl glucoside extraction of washed membrane vesicles in the presence of 20% glycerol and asolectin followed by ammonium sulfate fractionation and sucrose density gradient centrifugation . The purified preparation was resolved into seven bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, corresponding to the five F1 subunits, alpha, beta, gamma, delta, and epsilon, and to the b and c subunits of the F0 . Two-dimensional sodium dodecyl sulfate-poly-acrylamide gel analysis revealed a candidate for the alpha subunit of F0 . The MgATPase activity of B . firmus OF4 F1F0 was barely detectable but could be stimulated, optimally more than 100-fold, by sulfite, methanol, and octyl thioglucoside . The enzyme was inhibited by N,N'-dicyclohexylcarbodiimide and sodium azide, but not by aurovertin, an inhibitor of the F1 from Escherichia coli . The F1F0 reconstituted into proteoliposomes catalyzed ATPase activity, ATP-Pi exchange, and ATP-dependent delta pH and delta psi formation . ATP hydrolysis was stimulated by protonophores while the other activities were abolished by protonophores . These activities were neither dependent on added sodium ions nor significantly affected by them . F1F0 proteoliposomes made from crude octyl glucoside extracts that also contained the Na+/H+ antiporter were shown to catalyze ATP-dependent Na+ uptake that was completely sensitive to carbonyl cyanide m-chlorophenyl-hydrazone; Na+ uptake activity was absent in proteoliposomes containing more purified F1F0 but lacking the Na+/H+ antiporter . These data show that the F1F0 translocates protons and does not substitute Na+ for H+ in energy coupling.

Am J Ophthalmol, 1990 Nov 15, 110(5), 522 - 6
A randomized study of methanol-extraction residue of bacille Calmette-Guerin as postsurgical adjuvant therapy of uveal melanoma; McLean IW et al.; A randomized controlled clinical trial of methanol-extracted residue of bacille Calmette-Guerin adjuvant treatment of posterior uveal melanoma was undertaken . Of 113 patients, 34 patients received adjuvant immunotherapy and 79 patients received no treatment . No difference in survival was observed between the adjuvant-treated group and the control group of patients . This study found that the size of the tumor was a highly significant risk factor for death caused by metastasis of uveal melanomas . The standard deviation of the nucleolar area of the neoplastic cells was a significant risk factor, even though patients with tumors composed of Callender's spindle-type cells were not included in the study.

FEBS Lett, 1990 Nov 12, 274(1-2), 57 - 60
Crystal structure of an alkaline protease from Bacillus alcalophilus at 2.4 A resolution; Sobek H et al.; The crystal structure of an alkaline protease from Bacillus alcalophilus has been determined by X-ray diffraction at 2.4 A resolution . The enzyme crystallizes in space group P2(1)2(1)2(1) with lattice constants a = 53.7, b = 61.6, c = 75.9 A . The structure was solved by molecular replacement using the structure of subtilisin Carlsberg as search model . Refinement using molecular dynamics and restrained least squares methods results in a crystallographic R-factor of 0.185 . The tertiary structure is very similar to that of subtilisin Carlsberg . The greatest structural differences occur in loops at the surface of the protein.

Immunopharmacology, 1990 Nov-Dec, 20(3), 183 - 6
In vivo augmentation of local xenogeneic graft-versus-host reaction by cimetidine and Bacillus Calmette-Guerin; Snyman JR et al.; Histamine-2 receptor blocker therapy (cimetidine) was combined with nonspecific Bacillus Calmette-Guerin (BCG) immunomoderation in order to obtain a more directed immunostimulatory response . To establish this the local xenogeneic graft-versus-host reaction (XGVHR) was used as objective in vivo parameter . In this placebo-controlled experiment the lymphocytes of inbred Sprague-Dawley rats were transferred intradermally to partially immunosuppressed guinea pigs after treatment with either or both of the test substances . The surface areas of the local XGVHRs were evaluated 48 and 72 h later . The local XGVHRs caused by lymphocytes obtained from rats treated with either cimetidine or BCG alone did not differ significantly from those caused by lymphocytes obtained from placebo-treated rats . Combination therapy, however, resulted in significant augmentation of the local XGVHR, clearly indicating a synergistic effect between cimetidine and BCG . The XGVHR appears to be a satisfactory model for evaluating drug effect on cell-mediated immune reaction in vivo.

Kekkaku, 1990 Nov, 65(11), 747 - 54
{Practical use of surveillance system information to tuberculosis control}; Ikari A; In Japan as a low prevalence country in tuberculosis, national computerized tuberculosis surveillance system in Japan has started since in 1987 . The volume of information has much increased in comparison with previous annual reports, and its practical use to tuberculosis control programme is more expected . In this symposium, I presented two analyses, utilizing the information obtained from the surveillance system in Aichi Prefectural Government . 1) Surveillance of chemoprophylaxis 805 children were registered for chemoprophylaxis for seven years from 1980 . This comprised of 19.0 percent of the average annual bacillary pulmonary cases . In this study, we chose 553 cases of them who were detected in contact examinations and/or in medical institutions . Their information was sought at health centers . The treatment periods were insufficient in 10 percent, and the infectious sources were detected in 80 percent . 50 percent of them satisfied the criteria for chemoprophylaxis . The effectiveness of chemoprophylaxis was confirmed, though 4 cases developed tuberculosis later . 2) Surveillance of microepidemics 632 so called "danger group" patients of, infants, school students, teachers and medical staffs, were registered during six years from 1981 . 71 contact surveys had been made . Survey was made more frequently when the index case was bacillary positive . 3 microepidemics were found in this 6 years . Coordination among health centers were needed in one third of them to make examinations.(ABSTRACT TRUNCATED AT 250 WORDS)

Appl Environ Microbiol, 1990 Nov, 56(11), 3420 - 8
Novel cloning vectors for Bacillus thuringiensis; Baum JA et al.; Seven replication origins from resident plasmids of Bacillus thuringienis subsp . kurstaki HD263 and HD73 were cloned in Escherichia coli . Three of these replication origins, originating from plasmids of 43, 44, and 60 MDa, were used to construct a set of compatible shuttle vectors that exhibit structural and segregational stability in the Cry- strain B . thuringiensis HD73-26 . These shuttle vectors, pEG597, pEG853, and pEG854, were designed with rare restriction sites that permit various adaptations, including the construction of small recombinant plasmids lacking antibiotic resistance genes . The cryIA(c) and cryIIA insecticidal crystal protein genes were inserted into these vectors to demonstrate crystal protein production in B . thuringiensis . Introduction of a cloned cryIA(c) gene from strain HD263 into a B . thuringiensis subsp . aizawai strain exhibiting good insecticidal activity against Spodoptera exigua resulted in a recombinant strain with an improved spectrum of insecticidal activity . Shuttle vectors of this sort should be valuable in future genetic studies of B . thuringiensis as well as in the development of B . thuringiensis strains for use as microbial pesticides.

Ophthalmology, 1990 Nov, 97(11), 1532 - 8
Retained intraocular foreign bodies and endophthalmitis; Mieler WF et al.; Retained intraocular foreign bodies (IOFBs) are associated with endophthalmitis in approximately 7 to 13% of cases . The role of prompt surgical removal of the foreign body along with the use of intravitreal antibiotics in reducing this figure is uncertain . Retained IOFBs presenting to The Medical College of Wisconsin between July 1986 and June 1989 were reviewed . A total of 27 cases were evaluated and surgically treated . None of the 27 cases presented with or developed clinical signs of endophthalmitis, yet bacterial cultures of the removed intraocular material were positive in seven cases (foreign body in 5 cases, the aqueous fluid and the vitreous fluid in 1 case each) . All eyes presenting within 24 hours of injury underwent immediate surgery (average, 4.5 hours after presentation) . Of the seven eyes with positive intraocular cultures, all had pars plana vitrectomy removal of the IOFB and three of these eyes received intravitreal antibiotics at the time of surgery over concern of a high risk of infection . Two of these eyes eventually grew out the Bacillus sp . All eyes received subconjunctival antibiotics and postoperative topical and systemic antibiotics . Even after the positive cultures, no signs of clinical infection developed in any of the eyes . All seven eyes retained excellent visual acuity of 20/70 or better at an average of 10 months' follow-up . Follow-up ranged from 1 to 31 months . Prompt surgical intervention, the use of intravitreal antibiotics in high-risk-type injuries, and the possible use of vitrectomy surgery may reduce the incidence and severity of endophthalmitis.

J Am Vet Med Assoc, 1990 Nov 1, 197(9), 1176 - 8
Tyzzer disease in hamsters and gerbils from a pet store supplier; Motzel SL et al.; An episode of Tyzzer disease (Bacillus piliformis) developed in hamster and gerbil colonies of a pet store supplier . The incidence of diarrhea and subsequent mortality was high . The only important necropsy findings were cecal distention and mesenteric lymphadenopathy in the hamsters . Histologically, necrotizing typhlitis and hepatitis with associated B piliformis organisms were seen in both species . This case was unusual because the most consistent gross lesion associated with Tyzzer disease--hepatomegaly with multiple pale foci of hepatic necrosis--was not seen . Tyzzer disease is widespread geographically and among species; B piliformis has been reported to cause disease in at least 18 species of animals including hamsters, gerbils, rabbits, guinea pigs, horses, cows, dogs, and cats . Clinical signs of disease are nonspecific, and treatment is difficult because the organism is intracellular, although tetracycline and oxytetracycline reportedly have controlled mortality.

J Invertebr Pathol, 1990 Nov, 56(3), 312 - 6
The fate of Bacillus thuringiensis var . israelensis in B . thuringiensis var . israelensis-killed pupae of Aedes aegypti; Khawaled K et al.; Carcasses of mosquito larvae killed by Bacillus thuringiensis var . israelensis allow its complete growth cycle (germination, vegetative growth, and sporulation), thus becoming toxic themselves to scavenging larvae . In this study, we demonstrate that the bacterium is capable of inducing death of Aedes aegypti pupae and of recycling in the resulting carcasses . B . thuringiensis var . israelensis-killed pupae were obtained by treating 40-hr-old synchronized fourth instar larvae with a low dose of spores (8000/ml) . The fraction of dead pupae was reduced by higher or lower spore concentrations as well as by treating younger or older larval populations (both fourth instar): Increased proportions of dead larvae were obtained at higher concentration or by earlier treatment, whereas lower concentrations or later treatment resulted in more living pupae . Multiplication of B . thuringiensis var . israelensis is shown to occur in the carcasses of dead pupae . The number of spores in each pupal carcass followed a similar kinetic as in larval carcasses, but the final yield was about 10-fold higher, apparently reflecting the difference in dry weight between the two mosquito developmental stages (426 micrograms vs 83 micrograms, respectively) . The specific larvicidal activity in a homogenized dead pupa was similar to that of B . thuringiensis var . israelensis powder, LC50 of about 600 spores/ml.

Am Rev Respir Dis, 1990 Nov, 142(5), 996 - 9
Six-month isoniazid-rifampin treatment for pulmonary tuberculosis in children; Reis FJ et al.; One hundred and seventeen children with pulmonary tuberculosis underwent treatment with a 6-month daily regimen of rifampin (15 mg/kg/day) and isoniazid (10 mg/kg/day) . The criteria for the diagnosis of pulmonary tuberculosis were (1) clinical symptoms and signs in 93 children (79%), (2) history of direct contact with an adult with tuberculosis in 106 children (91%), (3) tuberculin reaction of 5 mm or more, without previous bacillus Calmette-Guerin (BCG), in 45 children (38%), (4) suggestive radiologic alterations in all patients, and (5) positive bacteriology or histology in four patients (3%) . The treatment was completed by 97 children (83%) . The mean weight gain during therapy was 2,145 g . There was an excellent clinicoradiologic response to the treatment, and improvement in chest roentgenograms was observed in all patients at the end of therapy . No relapses occurred among the patients followed for an average of 21.4 months . This study indicates that the treatment of primary pulmonary tuberculosis in children with a combination of rifampin and isoniazid daily for 6 months is efficacious and does not result in any relapse.

J Clin Oncol, 1990 Nov, 8(11), 1858 - 67
Treatment of metastatic melanoma with an autologous tumor-cell vaccine: clinical and immunologic results in 64 patients; Berd D et al.; We treated 64 patients with metastatic melanoma using a melanoma vaccine preceded by low-dose cyclophosphamide (CY), and monitored immunologic effects and antitumor activity . On day 0, the patients were given CY 300 mg/m2 intravenously . Three days later, they were injected intradermally with vaccine consisting of 10 to 25 x 10(6) autologous, enzymatically dissociated, cryopreserved, irradiated (25 Gy) tumor cells mixed with bacillus Calmette-Guerin (BCG) . This treatment sequence was repeated every 28 days . Of 40 assessable patients with measurable metastases, five had responses, four complete and one partial, with a median duration of 10 months (7 to 84+ months) . In six additional patients, we observed an antitumor response that seems to be peculiar to this vaccine therapy: the regression of metastatic lesions that appeared after the immunotherapy was begun . Delayed-type hypersensitivity (DTH) to autologous, mechanically dissociated melanoma cells that had not been exposed to extraneous antigens, such as enzymes or fetal calf serum, increased significantly following immunotherapy (day 0 v day 49, P less than .001; day 0 v day 161, P less than .001; day 0 v day 217, P = .021) . Antitumor responses to the vaccine were strongly associated with DTH, as indicated by three observations: (1) eight of 10 patients who exhibited tumor regression had positive DTH, (2) in postsurgical adjuvant patients, there was a highly significant linear relationship (P less than .001) between the intensity of DTH to autologous melanoma cells and the time to recurrence of tumor, and (3) nine patients who developed DTH to the autologous melanoma cells in their original vaccine developed new metastases that failed to elicit DTH or elicited a much smaller response . In three cases, we were able to excise regressing tumors for histologic examination; such tumors were characterized by an intense infiltration of lymphocytes . This demonstration that an immune response to melanoma-associated antigens can be elicited in cancer-bearing patients provides some basis for optimism about the prospects for developing active immunotherapy that has practical therapeutic value.

J Exp Med, 1990 Nov 1, 172(5), 1325 - 30
Suppression of developmental anomalies by maternal macrophages in mice; Nomura T et al.; We tested whether nonspecific tumoricidal immune cells can suppress congenital malformations by killing precursor cells destined to cause such defects . Pretreatment of pregnant ICR mice with synthetic (Pyran copolymer) and biological (Bacillus Calmette-Guerin) agents significantly suppressed radiation- and chemical-induced congenital malformations (cleft palate, digit anomalies, tail anomalies, etc.) . Such suppressive effects were associated with the activation of maternal macrophages by these agents, but were lost either after the disruption of activated macrophages by supersonic waves or by inhibition of their lysosomal enzyme activity with trypan blue . These results indicate that a live activated macrophage with active lysosomal enzymes can be an effector cell to suppress maldevelopment . A similar reduction by activated macrophages was observed in strain CL/Fr, which has a high spontaneous frequency of cleft lips and palates . Furthermore, Pyran-activated maternal macrophages could pass through the placenta, and enhanced urethane-induced cell killing (but not somatic mutation) in the embryo . It is likely that a maternal immunosurveillance system eliminating preteratogenic cells allows for the replacement with normal totipotent blast cells during the pregnancy to protect abnormal development.

Appl Environ Microbiol, 1990 Nov, 56(11), 3505 - 10
Purification and characterization of thermostable beta-mannanase and alpha-galactosidase from Bacillus stearothermophilus; Talbot G et al.; Bacillus stearothermophilus secretes beta-mannanase and alpha-galactosidase enzymatic activities capable of hydrolyzing galactomannan substrates . Expression of the hemicellulase activities in the presence of locust bean gum was sequential, with mannanase activity preceding expression of alpha-galactosidase activity . The hemicellulase activities were purified to homogeneity by a combination of ammonium sulfate fractionation, gel filtration, hydrophobic interaction chromatography, and ion-exchange and chromatofocusing techniques . The purified beta-D-mannanase is a dimeric enzyme (162 kilodaltons) composed of subunits having identical molecular weight (73,000) . Maximal activity did not vary between pH 5.5 and 7.5 . The beta-D-mannanase activity exhibited thermostability, retaining nearly full activity after incubation for 24 h at 70 degrees C and pH 6.5 . The enzyme displayed high specificity for galactomannan substrates, with no-secondary xylanase or cellulase activity detected . Hydrolysis of locust bean gum yielded short oligosaccharides compatible with an endo mode of substrate depolymerization . Initial rate velocities of the mannanase activity displayed substrate inhibition and yielded estimates for Vmax and Km of 455 +/- 60 U/mg and 1.5 +/- 0.3 mg/ml, respectively, at 70 degrees C and pH 6.5 . The alpha-galactosidase activity corresponded to a trimeric enzyme (247 kilodaltons) having subunits of identical molecular weight (82,000) . The alpha-galactosidase had maximal activity at pH 7 to 7.5 and retained full activity after 24 h of incubation at 60 degrees C . The enzyme had only limited activity on galactomannan substrates as compared with hydrolysis of p-nitrophenyl alpha-D-galactose . Kinetics of p-nitrophenyl alpha-D-galactose hydrolysis yielded linear reciprocal plots corresponding to Vmax and Km of 195 +/- 10 U/mg and 0.25 +/- 0.02 mM, respectively, at 60 degrees C and pH 7 . The characterization of the mannanase activity is consistent with its potential use in enzymatic bleaching of softwood pulps.

Cancer Res, 1990 Nov 1, 50(21), 6966 - 70
Evidence for the production of nitric oxide by activated macrophages treated with the antitumor agents flavone-8-acetic acid and xanthenone-4-acetic acid; Thomsen LL et al.; Activated peritoneal macrophages, obtained from mice pretreated with Bacillus Calmette-Guerin, after exposure in vitro to flavone-8-acetic acid (FAA; NSC 347512) at a concentration of 890 microM, produce nitrite (3.7 nmol/10(6) cells), as measured 20 h later by the Griess reaction . Stimulation of nitrite production was inhibited at least 90% by NG-monomethylarginine (125 microM), suggesting that nitrite was formed via nitric oxide as a product of arginine metabolism . Stimulation was only partially inhibited by dexamethasone (0.1 microM) . The ability of xanthenone-4-acetic acid (XAA) and three of its analogues to stimulate nitrite production was also investigated . 5,6-Dimethyl-XAA stimulated nitrite production (12.6 nmol/10(6) cells) at an optimal concentration of 80 microM, 8-methyl-XAA was without effect, and XAA and 5-methyl-XAA showed intermediate activity . The optimal in vitro drug concentrations for stimulation by FAA, XAA, and active XAA analogues correlated with the optimal in vivo dose required for the induction of either hemorrhagic necrosis or growth delay of s.c . Colon 38 tumors . These results strongly imply that FAA and active XAA derivatives function as low molecular weight stimulators of nitric oxide formation in macrophages, possibly acting on the same differentiation pathway as do endotoxin and tumor necrosis factor alpha . We suggest that nitric oxide, which is known to be toxic to tumor cells, contributes to the cytotoxic action of FAA and its analogues.

Plasmid, 1990 Nov, 24(3), 190 - 4
Presence of the bacterial hemoglobin gene improves alpha-amylase production of a recombinant Escherichia coli strain; Khosravi M et al.; A recombinant plasmid (pMK57) was constructed by cloning the Bacillus stearothermophilus alpha-amylase gene into pUC8; plasmid pMK79 was then derived from pMK57 by inserting the bacterial (Vitreoscilla) hemoglobin gene into the latter plasmid . Both pMK57 and pMK79 were transformed into Escherichia coli strain JM 103 to make strains MK57 and MK79, respectively . Both MK57 and MK79 produced alpha-amylase and MK79 produced hemoglobin . MK79 outgrew MK57 in shake flasks in LB medium, the advantage of the former appearing in late log phase . MK79 produced more alpha-amylase than MK57, on both per cell and per volume bases, in both mid and late log phases; the maximum advantage of MK79 (on a per volume basis) occurred in late log phase, at which time it produced 3.3 times as much alpha-amylase as MK57 . The numbers of copies per cell of both pMK57 and pMK79 were significantly lower than that of pUC8.

Rev Hosp Clin Fac Med Sao Paulo, 1990 Nov-Dec, 45(6), 248 - 52
{Contamination of enteral diet solutions in nosocomial environment}; Faintuch J et al.; The gastrointestinal tract is comparatively resistant to food-born germs, but recent studies suggest that nosocomial infections may be triggered by this route . In a study with industrialized diets, prepared with aseptic technique and stored up to 24 hours, aerobic and anaerobic contaminants were searched . Samples were taken after 0.8 and 24 hours, whereas half of these last two analyses were carried out in material left at room temperature, and the other half in refrigerated diets . Initial examination revealed 50% of positive cultures, but part of this was due to non-pathogenic Bacillus germs . After 8 and 24 hours 90% of the samples grew organisms, again with a large proportion of Bacillus, but also with several Gram-negative bacteria, as well as rare Gram-positives . Diarrhea and fever were not registered in patients submitted to enteral nutrition during the study period, nor could any episodes of bacteremia or septic shock be attributed to contaminated feeding material . This lack of clinical consequences of the reported bacterial isolations is not unexpected, and suggests that low concentrations of microorganisms were probably present in the preparations, below a critical level . Nevertheless, attention will be required in the future for better quality control of enteral nutrition mixtures, specially when resistant strains of Gram-negative species are identified, and also in the management of debilitated or immunologically compromised hosts.

Eur J Clin Microbiol Infect Dis, 1990 Nov, 9(11), 830 - 2
Degranulation of human neutrophils after exposure to bacterial phospholipase C; Wazny TK et al.; Because an endogenous phospholipase C (PLC) participates in neutrophil activation and because many bacterial pathogens produce PLCs, these studies examined the effect of PLC from Bacillus cereus on the release of the granule enzyme lysozyme from human neutrophils . Bacillus cereus PLC caused dose-dependent lysozyme release, and combined stimulation of neutrophils with PLC and fluoride led to increased secretion . Stimulation of neutrophil degranulation is a potential contributing factor for tissue damage in infections caused by PLC-producing organisms.

Rev Clin Esp, 1990 Nov, 187(7), 329 - 33
{Morphological evidence of the presence of a bacillus in cell cultures of the synovial membrane from rheumatoid arthritis patients}; Muelas Medrano JM et al.; 34 synovial membranes (SM) from 24 rheumatoid arthritis (RA) patients, and two from osteoarthrosis cases as controls, were studied by culturing the cells on elongated coverslips and Giemsa staining them . 33/34 RA-SM, a 97%, disclosed the presence of a rod-shaped bacterium and its cell wall-deficient forms (CWDB), all adhering to the cells . An external contamination as the organism origin, was ruled out . From one of the RA-SM, a Gram (-), nonfermentative bacillus was isolated, which up to now remains unidentified . This bacillus, when reinoculated onto human embryonic fibroblasts, reproduces the images shown in the rest of the RA-SM . The possibility of this bacillus being the etiological agent of RA is discussed.

FEMS Microbiol Lett, 1990 Nov, 60(3), 265 - 73
An analysis of the genes encoding the 51.4- and 41.9-kDa toxins of Bacillus sphaericus 2297 by deletion mutagenesis: the construction of fusion proteins; Oei C et al.; A series of deletion mutants have been constructed, in which varying numbers of amino acids have been deleted from both the N- and C-termini of both the 51.4- and 41.9-kDa toxins of Bacillus sphaericus . The results show that between 34-39 and 52-54 amino acids respectively at the N- and C-termini of the 51.4-kDa protein, are not essential for toxicity . In the case of the 41.9-kDa protein, the removal of only 7 amino acids from the C-terminus abolishes toxicity whilst at least 17 amino acids can be deleted from the N-terminus without loss of toxicity . A fusion protein with the 51.4-kDa derived sequence N-terminal to the 41.9-kDa sequence yielded a protein of Mr 87 kDa which was not toxic by itself . When supplemented with cells expressing only the 51.4-kDa protein, toxicity was restored . In contrast, another fusion protein, in which the gene order was reversed, was shown to be fully active in toxicity assays.

Mol Microbiol, 1990 Nov, 4(11), 1967 - 73
Larvicidal activity of chimeric Bacillus thuringiensis protoxins; Raymond KC et al.; Bacillus thuringiensis subspecies kurstaki (Btk) and subspecies berliner (Btb) both produce lepidopteran-specific larvicidal protoxins with different activities against the same insect species . Toxic activity resides in the amino-terminal half of both protoxins, whereas the carboxy-terminal half of the molecules is not required for toxicity . The protoxins are 90% homologous, with a major cluster of differences in the amino-terminal half, and a 26 consecutive amino-acid insertion within the carboxy-terminal half of the Btk protoxin . Protoxin chimeras composed of the amino-terminal half of one subspecies and the carboxy-terminal half of the other were generated . Wild-type and chimeric protoxins were compared in bioassays against tobacco hornworm larvae . The amino-terminal half, the toxin itself, dictates specific larvicidal activity.

Infection, 1990 Nov-Dec, 18(6), 388 - 93
Plague and Glasnost . First information about human cases in the USSR in 1989 and 1990; Velimirovic B; In October 1989 the first case of plague death in the USSR was reported to WHO . This occurrence in man did not surprise plague experts . The country has extensive enzootic areas and the persistence of natural foci, which can be silent for many years, has been well studied . It is known that the plague bacillus can survive and multiply in the soil of rodent burrows and restart local or more extensive transmissions in carrier animals . Isolated cases in man can remain accidental or they may signal a larger epizootic outbreak . The official policy of the comprehensive antiplague services was to eradicate the natural foci by antirodent activities which proved impossible . The present report from the Central Asian part of the USSR in the wake of Glasnost augurs well for the surveillance of plague worldwide as for a period of over fifty years the occurrence of cases in man in this country had been denied.

Can J Microbiol, 1990 Nov, 36(11), 804 - 7
Role of the gut proteinases from mosquito larvae in the mechanism of action and the specificity of the Bacillus sphaericus toxin; Nicolas L et al.; Gut proteinases from larvae of mosquito species both susceptible and not susceptible to Bacillus sphaericus converted the 43-kDa toxin to a 40-kDa polypeptide exhibiting enhanced cytotoxicity to mosquito cell cultures . The toxin was also activated by gut proteinases from the nonsusceptible Lepidoptera Spodoptera littoralis in vitro and in vivo . Therefore, the specificity of Bacillus sphaericus toxin does not seem to be determined by gut proteinase action . However, susceptibility of mosquito cell cultures did not reflect the specificity of the toxin, which must now be investigated at the cellular level in the larvae.

Biochemistry, 1990 Oct 23, 29(42), 9962 - 70
Asymmetric short-chain phosphatidylcholines: defining chain binding constraints in phospholipases; Lewis KA et al.; Several short-chain asymmetric lecithins with a total of 14 carbons in the acyl chains (ranging from 1-lauroyl-2-acetylphosphatidylcholine to 1-hexanoyl-2-octanoylphosphatidylcholine) have been synthesized and characterized . The specific activities of phospholipase A2 from cobra venom, phospholipase A2 from porcine pancreas, and phospholipase C from Bacillus cereus toward these lecithins as micelles have been determined . The results of these kinetic studies allow the definition of hydrophobic binding requirements in the active sites of these water-soluble phospholipases . For phospholipase C, with the exception of monomyristoylphosphatidylcholine, each of the asymmetric short-chain lecithins exhibits high activity, comparable to the 14-carbon symmetric short-chain species, diheptanoylphosphatidylcholine . Therefore, for phospholipase C, in addition to the acyl linkages, only a certain degree of hydrophobicity in the fatty acyl chains is requisite for substrate binding and appreciable hydrolysis; there is no chain specificity . The activity of phospholipase A2 from cobra venom toward the same asymmetric lecithins is quite different . As the sn-2 chain lengthens, activity is increased to a maximum for diheptanoyl-PC . Further increase in the number of carbons in the sn-2 chain has no effect on hydrolysis rates . For this enzyme, seven carbons in the sn-2 chain are necessary for optimal activity . In contrast, porcine pancreatic phospholipase A2 activity shows very little dependence on sn-2 chain length.

Biochim Biophys Acta, 1990 Oct 22, 1047(1), 77 - 82
Role of anionic lipid in bacterial membranes; Card GL et al.; The major phospholipids of Bacillus stearothermophilus are phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and cardiolipin (CL) . Under the growth conditions used in this study the concentration of anionic lipid (PG + CL) was determined by the pH of the culture medium . Cells grown in a complex medium at pH 5.8, 7.0, and 8.0 contained 17, 29 and 36 nmol of anionic (PG + CL) lipid/mg cell (dry weight) . The concentration of the zwitterionic lipid phosphatidylethanolamine (PE) was 17-20 nmol/mg cell (dry weight) under all conditions . Analysis of isolated membrane preparations suggested that the amount of anionic lipid per unit area of membrane increased as the pH of the growth medium was increased . Membranes from cells grown at pH 5.8 and 8.0 contained 130 and 320 nmol anionic lipid/mg membrane protein, respectively . Phosphatidylethanolamine appeared to be localized on the inner membrane surface in cells grown under all conditions . Increasing the ionic strength of the culture medium by the addition of NaCl or KCl had little effect on growth at pH 5.8 but inhibited growth at pH 7 and 8 . It was concluded that anionic phospholipid plays an important physiological role in maintaining an acidic pH at the outer membrane surface.

Biochim Biophys Acta, 1990 Oct 22, 1047(1), 41 - 8
Inhibition of the phosphatidylinositol-specific phospholipase C from Bacillus cereus by a monoclonal antibody binding to a region with sequence similarity to eukaryotic phospholipases; Kuppe A et al.; Bacterial phosphatidylinositol-specific phospholipases C (PI-PLC) display similar substrate specificity as their eukaryotic counterparts involved in signal transduction of insulin and Ca2(+)-mobilizing hormones, and are used in the study of the novel glycosylphosphatidylinositol-protein anchors (GPI-anchors) . For the investigation of structure-function aspects of the PI-PLC secreted from Bacillus cereus cells, a panel of murine monoclonal antibodies was generated and shown to be specific for the PI-PLC polypeptide in enzyme-linked immunosorbent assays and Western blots . Two of the monoclonals inhibited reactions catalyzed by the bacterial enzyme in vitro: hydrolysis of phosphatidylinositol and the release of bovine erythrocyte acetylcholinesterase from its GPI-anchor . At saturating concentrations of inhibitory antibody only a few percent of the enzyme activity remained . The epitope recognized by one of the inhibitory antibodies, A72-24, was mapped by proteolytic digestion, protein sequencing, and Western blotting of the generated fragments . The data indicate that at least part of the epitope resides within an 8 kDa-stretch of the bacterial PI-PLC (Gln-45 - Lys-122) . Essentially the same segment of the bacterial polypeptide has previously been shown to display limited amino acid sequence similarity with several eukaryotic PI-specific phospholipases C (Kuppe, A., Evans, L.M., McMillen, D.A . and Griffith, O.H . (1989) J . Bacteriol . 171, 6077-6083) . The results reported here suggest that the conserved peptide of these enzymes may contain functionally important residues.

Biochim Biophys Acta, 1990 Oct 12, 1036(1), 1 - 5
Purification and some properties of cyclodextrin-hydrolyzing enzyme from Bacillus sphaericus; Oguma T et al.; An intracellular cyclodextrin-hydrolyzing enzyme from Bacillus sphaericus E-244 isolated from soil was purified to a homogeneous state by means of Triton X-100 extraction, DEAE-Sepharose column chromatography, hydrophobic and molecular-sieve HPLC . The enzyme was estimated to have an Mr of 72,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 144,000 by HPLC gel filtration on TSK gel G 3000 SW . It had a pH optimum of 8.0, and the enzyme, stable at 25 degrees C and pH 5.5-9.5 for 24 h, was inactivated at 50 degrees C for 10 min . The enzyme hydrolyzed beta-cyclodextrin more effectively than linear maltooligosaccharides such as maltopentaose, maltohexaose and maltoheptaose or polysaccharides such as starch, amylopectin, amylose and pullulan.

Nucleic Acids Res, 1990 Oct 11, 18(19), 5807 - 10
II-Q restriction endonucleases--new class of type II enzymes; Degtyarev SK et al.; Unique restriction endonucleases Bpu 10l and Bsil have been isolated from Bacillus pumilas and Bacillus sphaericus, respectively . The recognition sequences and cleavage points of these enzymes have been determinated as 5'-CC1TNAGC-3'/3'-GGANT1CG-5' for Bpu 10l and 5'-C1TCGTG-3'/3'-GAGCA1C-5' for Bsil . Restriction endonucleases Bpu 10l and Bsil represent a new class of enzymes which recognize non-palindromic nucleotide sequences and hydrolize DNA within the recognition sequence . Bpu 10l and Bsil recognition sequences may be regarded as quasipalindromic and the enzymes may be designated as type II-Q restriction endonucleases.

Carbohydr Res, 1990 Oct 10, 206(2), 257 - 67
Studies of the mechanism of the cyclisation reaction catalysed by the wildtype and a truncated alpha-cyclodextrin glycosyltransferase from Klebsiella pneumoniae strain M 5 al, and the beta-cyclodextrin glycosyltransferase from Bacillus circulans strain 8; Bender H; The actions of the wildtype and a truncated alpha-cyclodextrin glycosyltransferase from Klebsiella pneumoniae strain M 5 al on malto-oligosaccharides showed no significant differences, and there was marked dependence of the kinetic parameters on the chain lengths of the substrate . The action of the beta-cyclodextrin glycosyltransferase from Bacillus circulans was less dependent on the chain length of the substrate, but Vmax of the initial cyclisation with the longer malto-oligosaccharides was only 28% of that determined for the enzyme of K . pneumoniae . The rate parameters suggested that the active site of each enzyme spans nine glucosyl residues, and that the catalytic sites are situated between subsites three and four for the K . pneumoniae enzymes and between subsites two and three for the B . circulans enzyme . The molecular binding affinities and the affinities of the 9th subsite were calculated from the rate parameters . The primary and tertiary structures of alpha-amylases and cyclodextrin glycosyltransferases are compared in the context of the reaction mechanism of the latter enzymes.

Biochemistry, 1990 Oct 9, 29(40), 9343 - 52
Estimating the contribution of engineered surface electrostatic interactions to protein stability by using double-mutant cycles; Serrano L et al.; Coulombic interactions between charges on the surface of proteins contribute to stability . It is difficult, however, to estimate their importance by protein engineering methods because mutation of one residue in an ion pair alters the energetics of many interactions in addition to the coulombic energy between the two components . We have estimated the interaction energy between two charged residues, Asp-12 and Arg-16, in an alpha-helix on the surface of a barnase mutant by invoking a double-mutant cycle involving wild-type enzyme (Asp-12, Thr-16), the single mutants Thr----Arg-16 and Asp----Ala-12, and the double mutant Asp----Ala-12, Thr----Arg-16 . The changes in free energy of unfolding of the single mutants are not additive because of the coulombic interaction energy . Additivity is restored at high concentrations of salt that shield electrostatic interactions . The geometry of the ion pair in the mutant was assumed to be the same as that in the highly homologous ribonuclease from Bacillus intermedius, binase, which has Asp-12 and Arg-16 in the native enzyme . The ion pair does not form a hydrogen-bonded salt bridge, but the charges are separated by 5-6 A . The mutant barnase containing the ion pair Asp-12/Arg-16 is more stable than wild type by 0.5 kcal/mol, but only a part of the increased stability is attributable to the electrostatic interaction . We present a formal analysis of how double-mutant cycles can be used to measure the energetics of pairwise interactions.

Radiology, 1990 Oct, 177(1), 77 - 81
Osteolytic lesions and bacillary angiomatosis in HIV infection: radiologic differentiation from AIDS-related Kaposi sarcoma; Baron AL et al.; Bacillary angiomatosis is a newly recognized multisystem bacterial infectious disease seen in the setting of human immunodeficiency virus (HIV) infection . The disease is marked by cutaneous vascular lesions that contain a bacterium similar to the cat scratch disease bacillus . Antibiotic therapy leads to the resolution of both cutaneous and systemic manifestations . Of 17 HIV-infected patients with cutaneous lesions of bacillary angiomatosis, six (35%) had symptomatic osteolytic bone lesions that improved following antibiotic therapy . The authors describe the appearance of the bone lesions on radiographs, computed tomographic (CT) scans, magnetic resonance (MR) images, and radionuclide studies . Osteolytic lesions are a relatively common feature of bacillary angiomatosis in patients with HIV infection . The presence of bone lesions aids in differentiation of bacillary angiomatosis from acquired immunodeficiency syndrome-related Kaposi sarcoma, which has similar cutaneous abnormalities but no associated bone lesions.

J Appl Bacteriol, 1990 Oct, 69(4), 550 - 6
Adhesion of bacillus spores in relation to hydrophobicity; Ronner U et al.; The adhesion of spores of five different Bacillus species to solid surfaces of different hydrophobicity was evaluated . The spore surface hydrophobicity was measured using hydrophobic interaction chromatography (HIC) . A large variation in hydrophobicity was found among the spores of the different species tested . The degree of adhesion of spores to the solid surfaces was consistent with the results obtained using the HIC method . The most hydrophobic spores, according to the HIC method, adhered in a much larger extent to the hydrophobic surfaces . Furthermore, spores generally adhered to a greater extent to hydrophobic and hydrophilic surfaces than did the vegetative cells.

Aust N Z J Med, 1990 Oct, 20(5), 702 - 4
Necrobacillosis--primary anaerobic septicaemia due to Fusobacterium necrophorum; Golledge CL et al.; Necrobacillosis is a severe septicaemic illness caused by an anaerobic Gram-negative bacillus, Fusobacterium necrophorum . It occurs predominantly in young people and is characterised by oropharyngeal infection and subsequent metastatic complications, frequently involving the lungs and/or large joints . We report four recent cases in previously healthy young men, describing the clinical features of the disease (including autopsy findings), and the problems associated with diagnosis and treatment.

J Econ Entomol, 1990 Oct, 83(5), 1813 - 7
Sprayable self-encapsulating starch formulations for Bacillus thuringiensis; McGuire MR et al.; Under glasshouse conditions, various cornstarches and adjuvants were examined as encapsulating agents in sprayable formulations for Bacillus thuringiensis subsp . kurstaki Berliner . When these materials were suspended in water and tested for physical factors such as viscosity, clumping, and retention on plant surfaces, a range of responses was observed . All formulations tested had acceptable viscosity, but some formulations clumped . When applied to cotton leaves, certain formulations remained less than 4 d, whereas others remained greater than 2 wk . One of these formulations containing equal parts of Mira-sperse (a commercially available pregelatinized cornstarch) and sucrose (6% total solids) was selected for testing persistence of B . thuringiensis insecticidal activity against Ostrinia nubilalis Hubner neonate larvae . When encapsulating agents were present, insecticidal activity decreased over a 2-wk period significantly more slowly than when encapsulating agents were absent . This indicates an enhanced persistence of B . thuringiensis under glasshouse conditions . Implications of a sprayable self-encapsulating formulation for B . thuringiensis are discussed.

Wei Sheng Wu Xue Bao, 1990 Oct, 30(5), 380 - 8
{Survey of Bacillus thuringiensis and Bacillus sphaericus from soils of four provinces of China and their principal biological properties}; Li R et al.; A number of isolates of Bacillus thuringiensis and Bacillus sphaericus were obtained from soils of Southwestern Area and Shaanxi Province of China . Among isolates of B . thuringiensis were under 13 sorts of serotype in total of 23 sorts of B . thuringiensis and about 20% of auto-agglutinate strains . Rules of ecologic distribution of two sorts of bacteria were analysed . Toxicities on six species of insects, morphology and crystal proteins of B . thuringiensis, as well as toxicities, morphology and crystal proteins of B . sphaericus, were investigated . 22 strains of more efficient of B . thuringiensis and 2 strains of more efficient of B . sphaericus were obtained . It was shown that B . thuringiensis is actually soil microorganism, and resource of B . thuringiensis is much fruitful in Southwestern Area of China.

Wei Sheng Wu Xue Bao, 1990 Oct, 30(5), 369 - 74
{Enzyme-linked immunosorbent assay of larvicidal toxic proteins of Bacillus sphaericus Ts-1}; Wang J et al.; Bacillus sphaericus Ts-1 Mosquito larvicidal toxins 42 k Da and 43 k Da were isolated by Sephadex G-200 chromatography . Three strains of highly toxic B . sphaericus and two non toxic strains were screened for toxic proteins using ELISA . The lowest detectable toxin level was 1.56 X 10(-5) mg/ml . Non toxic strains did not produce antigens reacting to either the 42 kDa or the 43 kDa antibodies . Ts-1 cultures were examined at 12 and 24 h by LC50 bioassay against Culex pipiens . The LC50's at 12 h and 24 h were 0.71 ppm and 0.154 ppm, respectively, i.e., the toxin level at 24 h was 4.6 times the level at 12 h . ELISA tests established total toxin at 0.049 mg/ml and 0.225 mg/ml at 12 h and 24 h, respectively, confirming the LC50 study.

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