Mikrobiologiia, 1986 Sep-Oct, 55(5), 825 - 30 {Ultrastructural and biochemical changes in Mycobacterium rubrum and Streptomyces bacillaris cells exposed to the herbicide semeron}; Poglazova MN et al.; The work was aimed at studying how the herbicide semerone affected the ultrastructure of two soil microorganisms, Mycobacterium rubrum and Streptomyces bacillaris . Depending on its concentration, the herbicide inhibited growth processes so that biomass yield decreased, cell division was interfered with, and giant and misshapen cells appeared . The herbicide taken at a concentration of 50-100 mg/ml increased the amount of membrane structures of the respiration type in some cells . This compound at a concentration of 400-500 mg/ml changed the nucleoid structure in certain cells . The decrease number of ribosomes and their peculiar distribution in the cell cytoplasm are most typical responses of the cells to the herbicide action . These responses were found in all cells at any of the tested herbicide concentrations . The results of cytological experiments are supported by statistically reliable data on the effect of the herbicide on RNA and protein synthesis . RNA synthesis is inhibited at a semerone concentration as low as 1 mg/ml, which is a very sensitive indicator of its presence in the medium. Biol Chem Hoppe Seyler, 1986 Sep, 367(9), 925 - 35 Isolation and identification of two protein-protein crosslinks within the two subunits of Bacillus stearothermophilus ribosomes; Brockmoller J et al.; Bacillus stearothermophilus 30S and 50S ribosomal subunits were isolated and crosslinked with diepoxybutane . The crosslinked proteins were extracted with LiCl or with 67% acetic acid and purified by a combination of different high performance liquid chromatography techniques . The protein fractions were analysed by two-dimensional and diagonal polyacrylamide gel electrophoresis and by immunological methods . Two crosslinked protein pairs, one from the large and one from the small subunit, consisting of proteins L23-L29 and S13-S19 respectively, were isolated in milligram amounts for determination of the crosslinked amino acids. J Clin Pathol, 1986 Sep, 39(9), 1021 - 4 Septicaemia and meningitis caused by dysgonic fermenter-2 (DF-2); Chan PC et al.; A patient developed dysgonic fermenter-2 (DF-2) septicaemia and meningitis after having been bitten by a dog . The pathogenic organism was isolated from the oral cavity of the dog, which, it is believed, is the first time that this has been done . The growth characteristics of the organism and the immunological response of the patient were studied . The isolation of the infecting agent from the dog confirms that the oropharyngeal flora of dogs is a reservoir of the organism: the fastidiousness and slow growth of this Gram negative bacillus may account for the small number of reported cases of infection caused by this organism. J Clin Microbiol, 1986 Sep, 24(3), 357 - 63 Spheroplastic phase of mycobacteria isolated from patients with Crohn's disease; Chiodini RJ et al.; Two strains of an unclassified Mycobacterium species were isolated after 18 and 30 months of incubation of media inoculated with resected intestinal tissues from patients with Crohn's disease . These strains represented the third and fourth isolates of this organism from Crohn's disease patients . Ultrastructural examination of this strain and two previously isolated strains revealed the presence of spheroplasts which eventually transformed into the bacillary form of a previously unrecognized Mycobacterium species . These cell wall-deficient forms did not stain with conventional dyes and failed to grow on hypertonic media . Restriction polymorphism of the ribosomal DNA genes was used to determine the relationship between the cell wall-deficient and bacillary forms . Identical restriction patterns of the ribosomal DNA genes were found between the spheroplasts and Mycobacterium sp . isolates with EcoRI, BamHI, and XhoI restriction endonucleases, thus providing definitive evidence of their origin . Unidentified spheroplasts were isolated from an additional 12 patients with Crohn's disease, of which 7 of 10 seroagglutinated with antiserum prepared against the Mycobacterium sp . Spheroplasts were isolated from 16 of 26 (61%) patients with Crohn's disease but not from tissues of 13 patients with ulcerative colitis or 13 patients with other diseases of the bowel . These findings support the role of mycobacteria as etiologic agents in some cases of Crohn's disease. Urology, 1986 Sep, 28(3), 173 - 5 Retrospective study of efficacy of intravesical BCG alone in treatment of superficial bladder cancer; Mydlo JH et al.; This is a review of 100 patients at our institution who were treated for superficial bladder cancer . In those patients with carcinoma in situ of the bladder who were treated with conventional therapy (resection and/or fulguration) and intravesical bacillus Calmette-Guerin (BCG) without intradermal BCG, and those patients who were treated with conventional therapy alone, we found a response rate of 60 per cent versus 40 per cent at the end of three months . In comparing those patients with superficial papillary cancer, we found a response of 39 per cent after conventional therapy and 63 per cent after conventional therapy and intravesical BCG . This suggests that intravesical BCG without intradermal BCG can be an important adjunct to the conventional therapy of bladder tumors. Equine Vet J, 1986 Sep, 18(5), 396 - 400 Elastase-producing microorganisms in horse lungs: their possible role in the pathogenesis of chronic pulmonary disease in the horse; Grunig G et al.; Seventeen out of 21 horses had pulmonary microbial organisms which reached considerable numbers in seven cases . Elastase-producing microorganisms from the environment (Streptomyces species and to a lesser extent Bacillus species) constituted 22 per cent to 99 per cent (mean 79 per cent) of the total growth . There was a considerable number of microorganisms with in vitro-produced elastases which were not or only slightly affected by horse serum . There was no correlation between numbers of organisms and pulmonary histopathological findings thus the significance of these microorganisms in the pathogenesis of alveolar emphysema is unknown . The growth of a strain of Streptomyces collinus/diastatochromogus isolated from the lungs was suppressed by fresh horse serum but not by decomplemented horse serum . Complement activation in response to this organism could contribute to airway inflammation through the production of mediators. J Exp Med, 1986 Sep 1, 164(3), 695 - 708 Immunoreactivity of a 70 kD protein purified from Mycobacterium bovis Bacillus Calmette-Guerin by monoclonal antibody affinity chromatography; Britton WJ et al.; The protein antigens from Mycobacterium bovis (BCG), M . tuberculosis, and M . leprae share a number of common determinants . We have used a murine mAb (L7) recognizing such a determinant on a protein of Mr 70,000 to purify this antigen from M . bovis sonicate by affinity chromatography . Enrichment of the protein in column eluates was confirmed by immunoblotting and in antigen inhibition assays . After radiolabelling with 125I, the protein could be immunoprecipitated with human lepromatous leprosy sera . Stimulation of peripheral blood mononuclear cells from BCG-vaccinated and naturally mantoux-positive individuals induced proliferation and IFN-gamma secretion, while intradermal injection of purified antigen into the same subjects resulted in a delayed-type hypersensitivity reaction . Thus, the 70,000 molecule carried epitopes capable of reacting with B cells, and eliciting a potentially protective T cell response . The first 15 N-terminal residues were sequenced using a gas-phase sequenator. J Am Mosq Control Assoc, 1986 Sep, 2(3), 325 - 8 Effects of sublethal exposure to Bacillus thuringiensis var . israelensis on larval development and adult size in Aedes aegypti; Hare SG et al.; The effects of exposure to sublethal concentrations of Bacillus thuringiensis var . israelensis (Serotype H-14) on second instar Aedes aegypti larvae were investigated . A test system was developed in which adverse effects would be detected as increased duration of larval development and decreased adult body size . No evidence of negative effects on survivors could be detected when sufficient B.t.i . dosages were applied to kill approximately half of the larvae in the treatment groups . However, when larval density was not controlled, and competition for food decreased as a result of larval mortality in the B.t.i.-treated groups, adult wing length was greater in the B.t.i . survivors than in the untreated controls . In addition, a residual mortality was noted in larvae that had been exposed to B.t.i . for 24 hr and then removed to habitats without B.t.i. Mycopathologia, 1986 Sep, 95(3), 133 - 8 A toxic metabolite of Nigrospora oryzae (Berk and Br.) petch; Wilson ME et al.; Nigrospora oryzae was isolated from dallisgrass (Paspalum dilatatum Poir.) collected in Auburn and from hay shipped under refrigeration to Florida . Some of these samples were eaten by cattle and horses that subsequently developed lameness . Metabolites of N . oryzae were separated by thin layer chromatography and tested for toxicity . Only one metabolite was toxic . Metabolite A showed toxicity to brine shrimp with an LD50 = 500 micrograms/ml in 8 h . It also had an antibiotic effect on Bacillus megaterium ATCC 14581 with a minimum inhibitory level of 10.1 micrograms/disc . As little as 435 micrograms of a crude methanolic extract of N . oryzae showed mild toxicity to chick embryos . The metabolite was not toxic to mice nor rats at the levels tested . Quantitative procedures developed for the determination of metabolite A showed that the maximum production occurred in yeast extract-sucrose liquid medium with an initial pH of 5-6, when incubated as a stationary culture for 28 days at 25 degrees C . It was concluded that metabolite A is a weak antibiotic rather than a mycotoxin, and was probably not associated with the symptoms of lameness observed in cattle and horses . The antibiotic is not one previously reported for N . oryzae. Infect Control, 1986 Sep, 7(9), 462 - 5 A hospital cafeteria-related food-borne outbreak due to Bacillus cereus: unique features; Baddour LM et al.; Although Bacillus cereus is a well-known cause of food-borne illness, hospital-related outbreaks of food-borne disease due to B . cereus have rarely been documented . We report a hospital employee cafeteria outbreak due to foods contaminated with B . cereus in which an outside caterer was employed to prepare the suspect meals . Data were collected from 249 of 291 employees who had eaten either of the two meals . With a mean incubation period of 12.5 hours, 64% (160 of 249) of employees manifested illness . Symptoms, which averaged 24.3 hours in duration, included diarrhea (96.3%), abdominal cramps (90%), nausea (50.6%), weakness (24.7%), and vomiting (13.8%) . Eighty-seven employees sought medical attention, 84 of whom were seen in an emergency room . Although a significant difference was not demonstrated in food-specific attack rates, B . cereus was cultured from both rice and chicken items that were served at both meals . Sixty-three employees submitted stools for culture that grew no enteric pathogens, but none were examined for B . cereus . This food-borne outbreak demonstrates: the need for hospital kitchen supervisors to ensure proper handling of food when outside caterers are employed; that significant differences in food-specific attack rates may not be demonstrated in outbreaks, which may be related to several factors; and the importance of notifying microbiology laboratory personnel when B . cereus is a suspect enteric pathogen, since many laboratories do not routinely attempt to identify this organism in stool specimens. Arch Biochem Biophys, 1986 Sep, 249(2), 588 - 95 Effects of metal ions on sphingomyelinase activity of Bacillus cereus; Ikezawa H et al.; Some divalent metal ions were examined for their effects on sphingomyelinase activity of Bacillus cereus . The enzyme activity toward mixed micelles of sphingomyelin and Triton X-100 proved to be stimulated by Co2+ and Mn2+, as well as by Mg2+ . Km's for Co2+ and Mn2+ were 7.4 and 1.7 microM, respectively, being smaller than the Km for Mg2+ (38 microM) . Sr2+ proved to be a competitive inhibitor against Mg2+, with a Ki value of 1 mM . Zn2+ completely abolished the enzyme activity at concentrations above 0.5 mM . The concentration of Zn2+ causing 50% inhibition of the enzyme activity was 2.5 microM . Inhibition by Zn2+ was not restored by increasing concentrations of Mg2+ when the concentration of Zn2+ was above 10 microM . Ba2+ was without effect . When sphingomyelinase was incubated with unsealed ghosts of bovine erythrocytes at 37 degrees C, the enzyme was significantly adsorbed onto the membrane in the presence of Mn2+, Co2+, Sr2+ or Ba2+ . Incubation with intact or Pronase-treated erythrocytes caused enzyme adsorption only in the presence of Mn2+ . In the course of incubation, the enzyme was first adsorbed on the membranes of intact bovine erythrocytes in the presence of Mn2+; then sphingomyelin breakdown proceeded with ensuing desorption of adsorbed enzyme . Hot-cold hemolysis occurred in parallel with sphingomyelin breakdown . In this case, the hydrolysis of membranous sphingomyelin as well as the initial enzyme adsorption took place in the following order: unsealed ghosts greater than Pronase-treated erythrocytes greater than intact erythrocytes. Am Rev Respir Dis, 1986 Sep, 134(3), 593 - 608 Small cell lung cancer; Iannuzzi MC et al.; Lung cancer is the predominant fatal neoplasm of our time, and SCLC, which accounts for about 25% of all lung cancer, if untreated results in death in about 3 months . Currently employed aggressive combination chemotherapy has allowed a 4- to 5-fold improvement in median survival over untreated patients . Ten to 20% of patients with limited disease can be expected to have a long-term (2-yr) survival . The majority of patients, however, have extensive disease . For these patients the median survival is about 7 months . Less than 2% survive 2 yr . During the last 10 yr, experience in the treatment of thousands of patients has been reported . These trials, using a large variety of drug combinations, doses, and schedules as well as multiple modalities including radiotherapy, surgery, and bone marrow transplantation, demonstrate that a plateau has been reached with our present therapeutic approach . The development of new effective therapeutic strategies as well as prevention of SCLC require a better basic understanding of the cellular pathophysiology of the disease . A consistent chromosomal abnormality has been associated with SCLC . This may provide new insight into predisposition and pathogenesis of SCLC . How this chromosomal abnormality relates to loss of control of cell growth is under intense investigation . Similarly, during the past 3 yr, the identification of growth regulatory oncogenes has greatly improved our understanding of malignancy . The discovery that metastatic cells escape immune surveillance has led to attempts at modulating antigenic expression . The modulation of cellular antigenic expression may facilitate the destruction of tumor cells by host defense mechanisms . The understanding of the genetic basis of drug resistance may lead to approaches that prevent or delay resistance . This century has witnessed the emergence of SCLC as an important fatal neoplasm . It has also been during this time that another, formerly dominant pulmonary condition, tuberculosis, has been controlled . The reduction of tuberculosis was accomplished by a combination of scientific understanding, beginning with the discovery of Koch's bacillus, and public health measures . Perhaps a similar parallel for SCLC as well as other forms of cancer will be written . Basic cellular investigations with the new tools of molecular biology as well as measures to control exposure to predisposing environmental factors such as component of cigarette smoke may one day lead to control of SCLC. J Am Mosq Control Assoc, 1986 Sep, 2(3), 276 - 9 Evaluation of methoprene, temephos and Bacillus thuringiensis var . israelensis against Coquillettidia perturbans larvae in Minnesota; Sjogren RD et al.; Temephos, Bacillus thuringiensis var . israelensis (B.t.i.) and methoprene were tested for larval control of Coquillettidia perturbans . Neither temephos nor B.t.i . treatments at their maximum recommended dosages consistently reduced larval numbers . Larval emergencies were reduced 99% in test plots treated with experimental, controlled release methoprene briquets (Altosid SR-10) . Breeding sites of Cq . perturbans in a 5,000 km2 area were treated using methoprene briquets in the 1984 season . Adult populations in a 7,700 km2 area were 2.5 times higher in untreated areas than treated areas . Methoprene can effectively control larval populations of Cq . perturbans. Mikrobiologiia, 1986 Sep-Oct, 55(5), 792 - 5 {Biosynthesis of the antibiotic gramicidin S by a Bacillus brevis culture associated with a change in growth kinetics}; Lykov VP et al.; The effect of changes in the kinetics of growth on gramicidin S biosynthesis by Bacillus brevis var . G.-B . was studied . The synthesis of gramicidin S is induced when the specific growth rate decreases, whatever is the factor causing this decrease . As was found experimentally, only cells of a certain age can synthesize gramicidin S. Am J Surg Pathol, 1986 Sep, 10(9), 595 - 610 Histology, immunohistochemistry, and ultrastructure of the verruga in Carrion's disease; Arias-Stella J et al.; Twenty-six verruga peruana nodules were studied . The presence of Factor VIII-related antigen and Ulex europaeus lectin binding, and the ultrastructural finding of rudimentary cell junctions and pinocytotic vesicles establish the endothelial character of the proliferating cells in the verruga nodules . Whereas superficial lesions could show an angiomatoid pattern, deep-situated nodules tended to present a compact type of growth . Electron-microscopic studies have shown that Bartonella bacilliformis was found abundantly in the extracellular spaces in the florid lesions and that no organisms were present in the late, resolving subcutaneous nodules . Although no true intracellular "viable" microorganisms were noted, pseudopods of cytoplasm entrapping one or two bacteria and surrounding matrix substance were seen often . The characteristics of cytoplasmic inclusions previously described in verruga cells as "chlamydozoa" were detailed . The ultrastructure of the inclusions corresponded to endothelial phagocytic cells in which complex invaginations of the cell surface had produced a labyrinth of interconnected channels and vacuoles containing degraded bacteria, extracellular matrix components, or both . We conclude that in light microscopy the finding of Rocha-Lima's inclusions is the only definite morphologic evidence of the presence of bartonella in verruga lesions. J Am Vet Med Assoc, 1986 Aug 15, 189(4), 440 - 1 Aerobic blood culturing in cows with coliform mastitis; Powers MS et al.; Aerobic culturing was performed on blood samples from 20 cows with coliform mastitis and signs of systemic illness, and on blood samples from 21 cows with mastitis caused by other agents . No bacteria other than the skin contaminant Bacillus sp were isolated from the blood. J Gen Microbiol, 1986 Aug, 132 ( Pt 8), 2387 - 95 Purification and characterization of the secreted alkaline phosphatase of Bacillus licheniformis MC14: identification of a possible precursor; Hulett FM et al.; The most abundantly secreted protein from Bacillus licheniformis MC14 is alkaline phosphatase (APase) . A twofold purification yields a homogeneous enzyme . No discernible chemical-physical differences in the secreted APase distinguish this enzyme from the cell-bound APase(s) except a 10-fold higher specific activity . During the growth phase in which the bacterium was secreting APase into the medium an inactive cytosol protein antigenically similar but 3000 Da heavier than the subunit of the mature secreted APase was immunoprecipitated from the cytosol . A pulse-chase experiment showed the kinetics of disappearance of this protein from the cytosol to be correlated with the appearance of the secreted APase in the medium, suggesting that it may be a precursor to the secreted APase. J Antibiot (Tokyo), 1986 Aug, 39(8), 1041 - 6 Bagougeramines A and B, new nucleoside antibiotics produced by a strain of Bacillus circulans . II . Physico-chemical properties and structure determination; Takahashi A et al.; Bagougeramines A and B obtained as sulfates were soluble in water and positive to Sakaguchi, chlorine-tolidine and ninhydrin color reactions . Their structures were determined by acid hydrolysis and spectroscopic analysis . Structurally they were closely related to gougerotin and they contained the guanidino-D-alanine instead of the serine residue in gougerotin . Bagougeramine B had the spermidine instead of the 6'-NH2 in structure of bagougeramine A. J Antibiot (Tokyo), 1986 Aug, 39(8), 1033 - 40 Bagougeramines A and B, new nucleoside antibiotics produced by a strain of Bacillus circulans . I . Taxonomy of the producing organism and isolation and biological properties of the antibiotics; Takahashi A et al.; A bacterial isolate from soil, designated as TB-2125 had a unique pattern of multiple resistance to aminoglycoside antibiotics (AG) and produced new nucleoside antibiotics . Taxonomic properties of this strain fell into those of Bacillus circulans, providing unique characteristics such as strict susceptibility to acidic pH, motility of colony as well as multiple AG-resistance . Two new antibiotics which were named bagougeramines A and B had a broad antimicrobial activity and a specific activity against the two spotted spider mite. Carbohydr Res, 1986 Aug 1, 150, 265 - 72 Structure of a rhamnan from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a; Christian R et al.; The structure of a glycan from the surface-layer glycoprotein of Bacillus stearothermophilus strain NRS 2004/3a has been studied by 1H- and 13C-n.m.r . spectroscopy . The results indicate the glycan to be a polymer of the trisaccharide repeating-unit ----2)-alpha-L-Rhap-(1----2)-alpha-L-Rhap-(1----3)-beta-L-++ +Rhap-(1----. DNA, 1986 Aug, 5(4), 305 - 14 Bacillus thuringiensis entomocidal protoxin gene sequence and gene product analysis; Wabiko H et al.; A 3778-bp DNA sequence of the insecticidal protoxin gene coding sequence and flanking regions from Bacillus thuringiensis subspecies berliner 1715 has been determined . The protoxin is composed of 1155 amino acids, deduced from the nucleotide sequence, and has a calculated molecular mass of 130,615 daltons . To determine the DNA portion that encodes toxicity, sequential deletions were constructed from the 3' end of the coding region using nuclease Bal-31 . Using these mutants in an insect bioassay, we found that an amino-terminal 612-amino-acid peptide is toxic, whereas, a 603-amino-acid peptide is not toxic to insects . Ninety percent of the amino acid residues were homologous to the protoxins from closely related subspecies kurstaki HD-1-Dipel and sotto . The differences occurred both in the amino-terminal half, or toxic portion, and in the carboxy-terminal half . These differences were clustered in several regions . From comparative analysis of subspecies berliner and kurstaki, we propose a model whereby the protoxin molecule is divided into distinct structural and functional domains . These domains may be responsible for the differences in specific toxicities and spectra of insect host range among these subspecies. J Biol Response Mod, 1986 Aug, 5(4), 288 - 93 Antitumor effect of formalin-fixed Toxoplasma gondii organisms on EL4 lymphoma in Toxoplasma-infected mice; Suzuki Y et al.; The antitumor effect of formalin-fixed Toxoplasma organisms (f-Tp) as an immunostimulant on EL4 lymphoma was examined in Toxoplasma-infected syngeneic female C57B1/6 mice . A potent antitumor effect, a marked suppression of tumor growth, as well as a prolongation of lifespan, was induced by an injection with 10(7) f-Tp in a mixture of 2.5 X 10(5) EL4 cells . The antitumor effect of f-Tp could be observed regardless of the time, route, or dose of infection . The peritoneal exudate cells induced by f-Tp in Toxoplasma-infected mice showed an antitumor activity when the cells were implanted with tumor cells in normal mice, indicating that the cells activated by f-Tp caused the antitumor effect of f-Tp . The effect of f-Tp on EL4 in Toxoplasma-infected mice was significantly stronger than that of 10(7) live bacillus Calmette-Guerin organisms (BCG) in BCG-sensitized mice. Clin Orthop, 1986 Aug, (209), 198 - 201 Multiple joint sepsis by Hemophilus influenza in an adult; Cohen MA et al.; The bacterium Hemophilus influenza, also known as the Pfeiffer bacillus, is generally regarded as a disease of infancy and early childhood . An occurrence in adulthood is rare . A 43-year-old woman developed septic arthritis of more than one joint caused by Hemophilus influenza . The infrequency of this infection and the degree of difficulty in diagnosis is confirmed by a review of the literature. Am J Dermatopathol, 1986 Aug, 8(4), 331 - 5 Fatal disseminated bacillus Calmette-Guerin infection and arrested growth of cutaneous malignant melanoma following intralesional immunotherapy; de la Monte SM et al.; An 84-year-old man with aggressive cutaneous malignant melanoma, which failed to respond to systemic chemotherapy, was electively treated with intralesional bacillus Calmette-Guerin (BCG) immunotherapy . He died with widespread miliary granulomas, most likely representing disseminated BCG infection . However, at autopsy, the patient was found to have a minimal tumor burden and only a single focus of visceral metastatic malignant melanoma . In view of the clinically documented aggressive nature of this patient's disease, the arrested growth of his tumors is best attributed to the beneficial effect of BCG immunotherapy. Am Rev Respir Dis, 1986 Aug, 134(2), 296 - 9 Granulomatous pneumonitis induced by bacille Calmette-Guérin in the mouse and its treatment with cyclosporin A; Takizawa H et al.; Granulomatous pneumonitis was induced intravenously by an injection of BCG, and changes in the population of cells from bronchoalveolar lavage (BAL) fluid were examined . Increased lymphocytes in BAL fluid, especially Lyt-1 positive T-lymphocytes, were observed after the development of granulomatous pneumonitis . Cyclosporin A (Cy A) administered for 5 days before and for 5 days after BCG injection clearly suppressed development of the granuloma . The BAL cell count and cell population became almost the same as those in naive animals . The increase in the number of Lyt-1 positive T cells was abrogated by Cy A treatment . These results suggested the important role of Lyt-1 positive T cells in the development of granulomas and the possible beneficial effect of Cy A in human granulomatous lung diseases. Eur J Biochem, 1986 Aug 1, 158(3), 505 - 10 A transcription terminator in the 5' non-coding region of the tyrosyl tRNA synthetase gene from Bacillus stearothermophilus; Waye MM et al.; The 5' non-coding region of the tyrosyl-tRNA synthetase gene (tyrS) of Bacillus stearothermophilus is 324 nucleotides long . It contains a premature terminator and a strong promoter: these were identified in vitro by RNA run-off experiments and in Escherichia coli by construction of specific mutants . The terminator consists of a stem and loop structure followed by the string of T residues characteristic of rho-independent termination . This is preceded by another stem and loop structure which may permit the formation of an anti-terminator . Neither the promoter nor the premature terminator appears to be regulated by the tyrosyl-tRNA synthetase in vitro or in E . coli. Eur J Biochem, 1986 Aug 1, 158(3), 497 - 504 Isolation and characterization of a 2.2-kb operon preceding the alpha-amylase gene of Bacillus amyloliquefaciens; Kallio P et al.; A DNA region of 2.8 X 10(3) base pairs (2.8 kb) upstream of the Bacillus amyloliquefaciens alpha-amylase gene has been isolated . This DNA gave rise to a 2.2-kb transcript . The 3' end of the transcript was mapped with S1 nuclease and shown to terminate 49 base pairs upstream of the -35 region of the alpha-amylase promoter . In B . subtilis minicells this 2.2-kb transcript coded for three different polypeptides, thus indicating a polycistronic operon-type structure . The location and the order of the polypeptides were established using DNA deletions . The joining of the 2.2-kb operon to the downstream alpha-amylase gene in the plasmid pUB110 did not have any significant effect on the level of expression of the alpha-amylase. Proc Natl Acad Sci U S A, 1986 Aug, 83(16), 5988 - 91 Leishmania and Trypanosoma surface glycoproteins have a common glycophospholipid membrane anchor; Bordier C et al.; The variant surface glycoprotein (VSG) of the African trypanosomes is the major membrane protein of the plasma membrane of the bloodstream stage of the parasite . It is anchored in the plasma membrane by a glycolipid covalently bound to the C-terminal amino acid of the protein . The VSG is released through the action of a phosphatidylinositol-specific phospholipase C that removes dimyristoylglycerol and exposes the carbohydrate antigenic determinant common to all VSGs . Promastigotes of Leishmania have a predominant surface glycoprotein, termed p63, that is anchored in the plasma membrane in a similar way . A water-soluble form of p63 can be generated through the action of phosphatidylinositol-specific phospholipase C from trypanosomes or from Bacillus cereus . Either treatment exposes on the Leishmania p63 an antigenic determinant recognized by antibody prepared against the trypanosomal crossreacting determinant . These findings indicate that p63 and VSG have a common membrane anchor and are structurally related. Biochem J, 1986 Aug 1, 237(3), 865 - 70 The use of inhibitors to identify early events during Bacillus megaterium KM spore germination; Foster SJ et al.; The germination response of spores of Bacillus megaterium KM, as measured by loss of A600, is more than 95% inhibited by 1 mM-HgCl2 . Two Hg2+-sensitive sites (referred to as 'sites I and II') have been identified during germination . Site I represents a pre-commitment event and can be protected from HgCl2 by 50 mM-D-alanine, whereas site II represents a post-commitment event and is not D-alanine-protectable . At 1 mM-HgCl2, 25% of the spore population becomes committed to germinate, but an A600 loss of less than 5% occurs . In this system, loss of heat resistance was associated with commitment, whereas selective cortex hydrolysis, release of pyridine-2,6-dicarboxylic acid, Zn2+ and soluble peptidoglycan, as well as loss of refractility, were identified as post-commitment events . The commitment event was reversibly inhibited by several proteinase inhibitors and a membrane bulking agent . A model of spore germination based on these results is presented. J Gen Microbiol, 1986 Aug, 132 ( Pt 8), 2329 - 35 Molecular cloning and nucleotide sequence of the alkaline cellulase gene from the alkalophilic Bacillus sp . strain 1139; Fukumori F et al.; The cellulase gene from the alkalophilic Bacillus sp . strain 1139 was cloned in Escherichia coli using pBR322 . Plasmid pFK1 was isolated from transformants producing cellulase, and the cloned cellulase gene was found to be in a 4 X 6 kb HindIII fragment . The cellulase gene was subcloned in a functional state on a 2 X 9 kb DNA fragment and its nucleotide sequence was determined . The coding sequence showed an open reading frame encoding 800 amino acids . The pFK1-encoded cellulase had the same enzymic properties as the extracellular cellulase produced by the alkalophilic Bacillus sp . strain 1139, but its Mr was slightly higher. J Bacteriol, 1986 Aug, 167(2), 544 - 50 Respiratory systems of the Bacillus cereus mother cell and forespore; Escamilla JE et al.; The respiratory systems of the mother cells and forespores of Bacillus cereus were compared throughout the maturation stages (III to VI) of sporulation . The results indicated that both cell compartments contain the same assortment of oxidoreductases and cytochromes . However membrane fractions from young forespores were clearly distinct from those of the mother cell, i.e., lower content of cytochrome aa3, lower cytochrome c oxidase activity, higher concentration of cytochrome o, and a lower sensitivity of the respiration to the inhibiting effect of cyanide . This suggests that the cyanide-resistant pathway contributes more importantly to forespore respiratory activity than to activity in the mother cell compartment . During the maturation stages, the forespore NADH oxidase activity declined faster than in the mother cells . Other activities studied decreased steadily in both cell compartments . These findings together with the analysis of the kinetics of NADH-dependent reduction of cytochromes in the mature spore membranes indicated an impairment of electron flow between NADH dehydrogenase and cytochrome b . This impairment could be overcome by the addition of menadione. J Cell Sci, 1986 Aug, 84, 221 - 36 Structurally related Bacillus thuringiensis delta-endotoxins display major differences in insecticidal activity in vivo and in vitro; Knowles BH et al.; Many strains within the 22 serotypes of Bacillus thuringiensis produce crystal delta-endotoxins with slight differences in their insecticidal toxicity spectrum in vivo . Since the basis of this specificity is unknown, we chose to compare the activity of delta-endotoxins from three strains: B . thuringiensis var . kurstaki HD-1, var . aizawai HD-249 and var . thuringiensis HD-350, both in vivo and on insect cell lines in vitro . Immunoblotting with antisera to activated var . kurstaki P1 lepidopteran toxin revealed antigenic cross-reaction with the 130 X 10(3) Mr toxin of var . aizawai, and with polypeptides of 130 and 138 (X 10(3)) Mr from var . thuringiensis . In addition, crystals from var . kurstaki and var . aizawai contained an antigenically related 63 X 10(3) Mr protein that did not cross-react with antisera to the 130 X 10(3) Mr component . Bioassays on Pieris brassicae larvae (Lepidoptera) and Aedes aegypti larvae (Diptera) indicated that the 130 X 10(3) Mr protein of var . kurstaki, and the 138 plus 130 X 10(3) Mr components of var . thuringiensis killed only P . brassicae, while the 130 X 10(3) Mr protein of var . aizawai and the 63 X 10(3) Mr proteins of var . aizawai and var . kurstaki were toxic to both P . brassicae and A . aegypti . Activation of the 130 and 138 (X 10(3)) Mr proteins of the three varieties of B . thuringiensis with insect gut proteases yielded active products of 50-60 (X 10(3)) Mr . Assay of these products on a range of lepidopteran and dipteran cell lines revealed very different toxicity spectra: var . kurstaki killed only one lepidopteran line, var . thuringiensis killed two lepidopteran lines, while var . aizawai was cytolytic to all of the lepidopteran and most of the dipteran cell lines tested, reflecting its broader spectrum in vivo . Thus we have shown that antigenic cross-reaction of B . thuringiensis delta-endotoxins does not necessarily imply a similar toxicity spectrum in vivo or in vitro. J Bacteriol, 1986 Aug, 167(2), 719 - 21 Temperate Bacillus bacteriophage SP16 genome is circularly permuted and terminally redundant; Parker AP et al.; The physical nature of temperate Bacillus bacteriophage SP16 DNA was analyzed by electron microscopy, exonuclease digestion, denaturation-renaturation experiments, and restriction enzyme analysis . The SP16 genome is a linear molecule 60.0 +/- 2.0 kilobases in length without cohesive ends . Electron micrographs of denatured and renatured SP16 DNA showed that the DNA is circularly permuted . The genome possesses terminal redundancy, as demonstrated by electron microscopy of exonuclease III-digested DNA. J Bacteriol, 1986 Aug, 167(2), 716 - 8 Transposition of Tn917 in Bacillus megaterium; Bohall NA Jr et al.; Transposon Tn917, carried on plasmid pTV1, was introduced into Bacillus megaterium and transposed efficiently and apparently randomly . Insertional mutations included at least eight different auxotrophic loci, two carbon source loci, and sporulation loci . One trp::Tn917 mutation was further verified as an insertion by both reversion and transduction. Biochemistry, 1986 Jul 29, 25(15), 4219 - 23 A spectral study of cobalt(II)-substituted Bacillus cereus phospholipase C; Bicknell R et al.; The coordination sphere of both the structural and catalytic zinc ions of Bacillus cereus phospholipase C has been probed by substitution of cobalt(II) for zinc and investigation of the resultant derivatives by a variety of spectroscopic techniques . The electronic absorption, circular dichroic, magnetic circular dichroic, and electron paramagnetic resonance spectra were found to be strikingly similar when cobalt(II) was substituted into either site and are consistent with a distorted octahedral environment for the metal ion in both sites . Octahedral coordination appears comparatively rare in zinc metalloenzymes but has been suggested for glyoxalase I {Sellin, S., Eriksson, L . E . G., Aronsson, A.-C., & Mannervik, B . (1983) J . Biol . Chem . 258, 2091-2093; Garcia-Iniguez, L., Powers, L., Chance, B., Sellin, S., Mannervik, B., & Mildvan, A . S . (1984) Biochemistry 23, 685-689}, transcarboxylase {Fung, C.-H., Mildvan, A . S., & Leigh, J . S . (1974) Biochemistry 13, 1160-1169}, and the regulatory binding site of Aeromonas aminopeptidase {Prescott, J . M., Wagner, F . W., Holmquist, B., & Vallee, B . L . (1985) Biochemistry 24, 5350-5356} . Phospholipase C is so far unique in having two such sites. J Biol Chem, 1986 Jul 25, 261(21), 9576 - 8 Protein engineering of homodimeric tyrosyl-tRNA synthetase to produce active heterodimers; Ward WH et al.; Heterodimers of tyrosyl-tRNA synthetase from Bacillus stearothermophilus have been produced by mutagenesis at the subunit interface . Oppositely charged groups have been engineered into the subunits so that they can form a complementary pair . Wild-type tyrosyl-tRNA synthetase is a symmetrical dimer in which the side chains of the 2 Phe-164 residues interact at the subunit interface . Phe-164 was mutated to Asp in tyrosyl-tRNA synthetase and to Lys in a truncated enzyme (des-(321-419)tyrosyl-tRNA synthetase) which lacks the two tRNA-binding sites, but which can catalyze pyrophosphate exchange . The size difference allows subunit association to be studied by gel filtration chromatography . These changes induce reversible dissociation from active dimers into inactive monomers at pH values which favor ionization at position 164 . A mixture of the two mutants near neutral pH is apparently fully active in pyrophosphate exchange and consists of a heterodimer of {Asp164}tyrosyl-tRNA synthetase and {Lys164}des-(321-419)tyrosyl-tRNA synthetase . Despite having only one binding site for tRNA, heterodimer has full aminoacylation activity at high concentrations of tyrosine . We have therefore produced a family of dimers that differ in stability near neutral pH . This novel approach using protein engineering allows specific dimerization of subunits of the same size that have different defined mutations, each subunit being tagged by the charge . Such hybrid proteins can be used to study subunit interaction. Nature, 1986 Jul 31-Aug 6, 322(6078), 462 - 4 Cloned suppressor T cells from a lepromatous leprosy patient suppress Mycobacterium leprae reactive helper T cells; Ottenhoff TH et al.; Leprosy is a chronic infectious disease caused by Mycobacterium leprae . A characteristic feature of the disease is its remarkable spectrum of clinical symptoms correlating with the cellular immune responsiveness of the patient . At one pole of this spectrum are tuberculoid patients displaying both acquired cell-mediated immunity and delayed type hypersensitivity against the bacillus . At the other pole are lepromatous patients which show a specific T-cell unresponsiveness against M . leprae . In between those two poles variable degrees of tuberculoid and lepromatous features may be seen in borderline leprosy patients . Thus far, studies on the mechanism of the antigen specific unresponsiveness in lepromatous leprosy have been contradictory and difficult to interpret, probably because of the use of heterogeneous cell populations in those experiments . We have now succeeded in cloning M . leprae stimulated T-helper (TH) as well as T-suppressor (TS) cells from a borderline lepromatous patient . The TS-clones of this patient specifically suppress responses of peripheral TH cells as well as TH clones induced by both M . leprae and other mycobacteria, but not unrelated antigen or mitogen . These TS cells also completely suppress TH cell responses against a M . leprae specific protein with a relative molecular mass of 36,000 (36K), suggesting the presence of a suppression inducing determinant on this 36K M . leprae protein. J Gen Microbiol, 1986 Jul, 132 ( Pt 7), 1951 - 8 Temperature-dependent plasmid integration into and excision from the chromosome of Bacillus stearothermophilus; Koizumi J et al.; A transformant of Bacillus stearothermophilus carrying a recombinant plasmid, pLP11 (9.5 MDa), on which the penicillinase gene (penP) and kanamycin resistance gene (kan) were located was subjected to mutagenesis, and a mutant plasmid (9.5 MDa; penP kan), designated pTRA117, was obtained . A transformant of B . stearothermophilus carrying pTRA117 could grow at 63 degrees C in medium containing kanamycin, whereas a transformant carrying pLP11 could not . Although pTRA117 was detected as covalently closed circular (ccc) DNA when it was extracted from transformants cultured at 48 degrees C, it was integrated into the host chromosome when the culture temperature was shifted up to 63 degrees C . If the culture temperature was lowered to 48 degrees C from 63 degrees C, a new plasmid (10.7 MDa; penP kan), designated pTRZ117, could be detected as ccc DNA; the size of this plasmid suggested that it was pTRA117 plus a 1.2 MDa DNA fragment of the host chromosome, and this was confirmed by Southern hybridization . pTRZ90 (7.9 MDa; kan) was constructed from pTRZ117 by the deletion of a 2.8 MDa DNA fragment that contained penP . Fresh transformants of B . stearothermophilus that carried either pTRZ117 or pTRZ90 could grow at 65 degrees C. Pediatr Pulmonol, 1986 Jul-Aug, 2(4), 202 - 5 Protective value of BCG vaccination in children in Bangkok, Thailand; Chavalittamrong B et al.; The protective effect of bacille Calmette-Guerin (BCG) vaccination against tuberculosis is controversial . In a study, 330 patients less than 12 years of age with tuberculosis, 52.1% of whom had had BCG vaccination, were compared with a control group of 1106 patients free of tuberculosis, 81% of whom had BCG vaccination . The occurrence of disseminated forms of tuberculosis, tuberculous meningitis, tuberculous peritonitis, and tuberculosis of bone and joints in BCG-vaccinated patients was quite low . With BCG vaccination, the incidence of the disseminated form of tuberculosis was significantly lower than that of pulmonary tuberculosis with pulmonary parenchymal lesions, primary pulmonary complexes, and pleural effusion . Tuberculous peritonitis was significantly less frequent than pulmonary tuberculosis with pulmonary parenchymal lesions, enlarged hilar glands, pulmonary primary complex, and pleural effusions . The study demonstrated that BCG gave an overall protective effect of 74% and that a major effect of this immunity was to produce a localized form of tuberculosis. J Androl, 1986 Jul-Aug, 7(4), 264 - 9 Reversibility of azoospermia induced by bacillus Calmette-Guerin; Naz RK et al.; A single intratesticular injection of bacillus Calmette-Guerin induces azoospermia within 3 to 6 weeks in a variety of animals without loss of androgens . After a period of azoospermia, the return of spermatogenesis was observed in dogs and monkeys . Seven dogs that showed aspermatogenesis after a local instillation of bacillus Calmette-Guerin (20-110 units per testis) again demonstrated spermatogenesis after a period of 153 to 325 days of azoospermia . Sperm counts and motility were restored to preimmunization levels within 6 weeks after initiation of spermatogenesis . In one dog, systemic treatment with antimycobacterial drugs helped to clear the bacillus Calmette-Guerin from the testis, thus hastening the reinitiation of spermatogenesis . In two monkeys injected with 110 and 160 units of bacillus Calmette-Guerin, azoospermia lasted up to 150 days . Subsequently, fertility was restored and the treated animals sired healthy offspring . A second injection of bacillus Calmette-Guerin of a lesser dose (5-20 units per testis) in these fertile dogs and monkeys again produced azoospermia . Histologically, the monkeys having a second period of azoospermia demonstrated aspermatogenic orchitis in the testis and leukocyte infiltration in the interstitium . A majority of the tubules had intact basement membranes, with a block of spermatogenesis at the spermatocyte stage . In dogs, the second azoospermia lasted for 113 to 277 days with a subsequent restoration of sperm counts and sperm motility to preimmunization levels . There were no circulating antisperm antibodies detected in sera of these animals during the azoospermic period or during its reversal . These observations suggest that the bacillus Calmette-Guerin-induced azoospermia is reversible, both in terms of sperm count and fertility. Medicine (Baltimore), 1986 Jul, 65(4), 218 - 25 Polymicrobial septicemia in the cancer patient; Elting LS et al.; The medical records of 507 patients with polymicrobial septicemia were examined to determine prognostic and descriptive factors . Over 50% of the episodes occurred in patients with solid tumors and 80% originated during hospitalization . Invasive procedures and immunosuppressive therapy frequently preceded development of polymicrobial septicemia, and infection was often accompanied by shock and pneumonia . A majority of infections were caused by at least 1 aerobic gram-negative bacillus (76%) and anaerobic infections were not infrequent . Overall response among these patients was 50%, with poorest response seen among patients with persistent neutropenia (25%), pneumonia (19%), and gram-negative bacillary infection (46%) . Therapy with an antibiotic regimen to which all causative organisms were sensitive was of greatest prognostic significance . Response to appropriate therapy was 58%, whereas only 10% of those who received inappropriate therapy were cured (p less than .0001). Am J Public Health, 1986 Jul, 76(7), 783 - 6 A case-control study to evaluate the effectiveness of mass neonatal BCG vaccination among Canadian Indians; Young TK et al.; This paper reports a case-control study to assess the protective effect of BCG (bacille Calmette-Guerin) vaccination among Indian infants in Manitoba, Canada . A record of past BCG vaccination was found in 49 per cent of the tuberculosis cases, compared to 77 per cent of the controls, yielding a relative risk of 0.30 . Stratified analysis, controlling for age, increased the relative risk to 0.39 (95% confidence interval 0.22 - 0.69) . The preventive fraction was 44 per cent . Non-differential misclassification of exposure status could have occurred; if this was adjusted for, the relative risk would be reduced . If only bacteriologically confirmed cases were analyzed, the age-adjusted relative risk was 0.27 . The protective effect of BCG vaccination in the newborn among Manitoba Indians is therefore at least 60 per cent . The implications for health policy in this population are further discussed. Mikrobiyol Bul, 1986 Jul, 20(3), 200 - 5 {Bacillus thuringiensis plasmids}; Cakmakci L; Several B . thuringiensis serotypes are used for the production of bacterial insecticides . The important ingradient of bacterial insecticide produced by B . thuringiensis is a parasporal crystal protein formed during sporulation . The bioinsecticide containing several B . thuringiensis serotypes have great potential in the control of certain insects, because of their high selectivity and the absense of any harmful effect on humans, plants, animals, polinators and predators . But the generation of cry- (non-virulent or acrystalliferous) strains during the growth in the medium are relatively high without mutagenic treatments . Plasmids can play role in this area . Because plasmids have been detected in Bacillus thuringiensis strains belonging to several serotypes and have not been found in certain cry- varyants . It is concluded that parasporal crystals are plasmid determined in B . thuringiensis varyants. Prikl Biokhim Mikrobiol, 1986 Jul-Aug, 22(4), 543 - 7 {Chemical mutagenesis and the use of indirect enzymatic criteria for the selection of virulent clones of Bacillus thuringiensis}; Slavnova VS et al.; The effect of the mutagene nitrosoguanidine (N-methyl-N'-nitro-N-nitrosoguanidine) on the growth of Bacillus thuringiensis subsp . galleriae st . 69/6 was being studied . It depends on the physiological state of the cells, dose of the mutagene, pH of the culture medium and exposition . Nitrosoguanidine was found to have the maximum mutagenic effect on the vegetative cells at pH 6.2 and on the spores at pH 5.6. J Antibiot (Tokyo), 1986 Jul, 39(7), 994 - 1000 Isolation of an aminoglycoside hypersensitive mutant and its application in screening; Numata K et al.; An aminoglycoside hypersensitive mutant, Kp-126, was isolated from the aminoglycoside-resistant strain, Kp-8, of Klebsiella pneumoniae through selection using sorbistin, a non-aminocyclitol-aminoglycoside antibiotic . The mutant Kp-126 was approximately 100-fold more sensitive to sorbistin than the parent strain Kp-8 . The mutant also showed hypersensitivity to various aminocyclitol-aminoglycoside antibiotics . K . pneumoniae Kp-126 was used in screening and a new aminoglycoside antibiotic, 3,3'-neotrehalosadiamine (BMY-28251), was discovered in the fermentation broths of soil isolate strain of Bacillus pumilus. Postgrad Med J, 1986 Jul, 62(729), 653 - 5 Tuberculous meningitis due to Mycobacterium bovis: a report of two cases; Wilkins EG et al.; Two Caucasian patients with bovine tuberculous meningitis are described . Classical Mycobacterium bovis was isolated from the cerebrospinal fluid on both occasions . Despite the elimination of cattle tuberculosis in this country, reactivated primary disease due to the bovine tubercle bacillus may still occur. J Bacteriol, 1986 Jul, 167(1), 18 - 24 Bacteriophage-resistant mutants of Bacillus thuringiensis with decreased virulence in pupae of Hyalophora cecropia; Heierson A et al.; Starting from a crystal-negative parental strain of Bacillus thuringiensis, we isolated certain bacteriophage-resistant mutants which showed decreased virulence in pupae of the cecropia moth (Hyalophora cecropia) . These strains (class I mutants) were highly pleiotropic and showed resistance to seven or eight different phages, sensitivity to methicillin, and loss of flagella . They were also more sensitive to cecropia immune hemolymph in vitro . In addition, the export of at least three proteins was reduced . Revertants (class II mutants) were sensitive to phages, virulent, and resistant to penicillin derivatives . One class II mutant was a complete revertant in all properties examined . The other class II mutant was an incomplete revertant still susceptible to immune hemolymph and with repressed export of proteins . Virulence was not coupled to phage resistance as such or to lack of flagella because other mutants affected in these properties were virulent . Other factors which could be excluded as causes of virulence were production of extracellular protease and hemolysin. Med Clin North Am, 1986 Jul, 70(4), 933 - 44 Considerations in the therapy of septic shock; Karakusis PH; In summary, gram-negative sepsis is unique among infectious illnesses in that it is a disorder that recruits endogenous physiologic processes to mediate tissue injury . This host damage frequently occurs in the absence of microbial invasion of affected organs . The resultant hypotension, coagulation defects, and organ dysfunction may be associated with serious morbidity or may contribute to mortality . Ultimately, however, mortality in patients with septic shock depends on the nature of the infectious process and the severity of the underlying illnesses . Unfortunately, attempts to aggressively treat septic patients with a formidable array of antimicrobial and pharmaceutical agents have not remarkably reduced mortality . Nor does it seem likely that future elucidation of the inflammatory mechanisms of sepsis will lead to the generation of therapeutic agents that will significantly improve survival . On the other hand, prophylactic or therapeutic modalities that deter colonization or invasion by pathogenetic organisms or that alter the ability of pathogens to evoke adverse host responses may be more likely to impact on the incidence and morbidity of gram-negative bacillary infections . Until modifications in the initial interactions of gram-negative pathogens with human hosts can be realized, the mortality of gram-negative sepsis is likely to remain high. Mikrobiologiia, 1986 Jul-Aug, 55(4), 601 - 6 {Effect of surface-active agents on the electrical properties of bacterial cells}; Fomchenkov VM et al.; The work was aimed at studying the effect of cationic, anionic and non-ionogenic surfactants on the frequency dependence of the electroorientation effect (EOE) and on the electrophoretic mobility (EPM) of rod-like bacteria . The character of concentration dependences was found to differ for EOE and EPM at a low frequency of the electric field (20 to 10(4) Hz) . Analysis of EOE changes at a high frequency (4 X 10(5) to 3 X 10(7) Hz) showed that anionic and non-ionogenic surfactants at a concentration up to 10(-3) M did not damage Escherichia coli, a Gram-negative bacterium, in contrast to Bacillus cereus, a Gram-positive bacterium . Cationic surfactants affected the cells of the both species . The optical properties of bacterial cells were found to change under the action of cationic surfactants. Avian Dis, 1986 Jul-Sep, 30(3), 574 - 9 An improved microbiological assay for chlortetracycline in avian plasma; Miller BL et al.; Bacillus cereus was used as the assay organism for the quantification of chlortetracycline (CTC) in avian plasma . Antibiotic medium #8 gave significantly larger zones of inhibition than nutrient agar 1.5% when used as the assay medium (P less than or equal to 0.05) . When the CTC concentration was measured in serum, citrated plasma, heparinized plasma, and oxalated plasma, no significant differences were found between the inhibition zone diameters produced on antibiotic medium #8 . However, there was a significant decrease in the zone diameters produced on this medium when citrated whole blood, oxalated whole blood, and heparinized whole blood were used instead of plasma . The length of incubation of assay plates was inversely related to the inhibition zone diameter, with an incubation time of 8 hr giving the largest and most distinct zones . Storing prepared assay plates at 4 C for 24 hr before use appeared to decrease the variability of the inhibition zone diameters . Storage of citrated plasma at 4 C for 48 hr and at -60 C for 9 days resulted in no significant decreases in CTC concentrations. Poult Sci, 1986 Jul, 65(7), 1281 - 6 An on-farm method for determination of sulfonamide drug residues in turkeys . 1 . An agar-diffusion analysis for sulfadimethoxine in whole blood; Murphy ML et al.; A simple and inexpensive sulfonamide-screening test was evaluated using turkeys . An agar-diffusion procedure was developed to estimate the levels of sulfonamides in the edible tissues of turkeys by determining the drug level in whole blood . The analysis was adapted for use on whole blood that was easily collected from live birds on the farm with minimal equipment and skill . This Whole Blood Sulfa Test (WBST) was quantified by the use of a standard curve and was successfully applied to on-farm use in the Pacific Northwest . Agar plates were prepared using fortified Mueller-Hinton medium . Bacillus megaterium spores were applied to the agar to form confluent growth, and paper discs (10 mm) were laid onto the agar . Whole blood was collected from commercial turkeys prior to marketing, and the blood was immediately applied to the test paper discs . After incubation, blood that contained sulfa inhibited bacterial growth around the disc, and the clear zones of inhibition were measured . The WBST was consistently accurate to 1.22 ppm, and sulfa levels were detected as low as .04 ppm . Results were attained in 12 hr and were relatively inexpensive at $3.00/test/flock. Appl Environ Microbiol, 1986 Jul, 52(1), 64 - 7 Red pigment in Bacillus megaterium spores; Mitchell C et al.; Bacillus megaterium QM B1551 spores contained a unique red pigment in their membranes that was not found in other species . This red pigment, presumably a carotenoid, was synthesized about the time of dipicolinic acid synthesis during sporulation and was associated with the forespores . A yellow pigment was synthesized during sporulation in rich medium and was found in the mother cell compartment . Although the yellow pigment was also associated with spores, it could be removed by two different extraction procedures without impairing germination; it was absent when sporulation occurred in a minimal medium . Although the yellow pigment of the mother cell appeared to be dispensable, the red pigment may serve a more critical function, such as membrane stabilization. J Bacteriol, 1986 Jul, 167(1), 168 - 73 Cloning and nucleotide sequencing of genes for small, acid-soluble spore proteins of Bacillus cereus, Bacillus stearothermophilus, and "Thermoactinomyces thalpophilus"; Loshon CA et al.; As found previously with other Bacillus species, spores of B . stearothermophilus and "Thermoactinomyces thalpophilus" contained significant levels of small, acid-soluble spore proteins (SASP) which were rapidly degraded during spore germination and which reacted with antibodies raised against B . megaterium SASP . Genes coding for a B . stearothermophilus and a "T . thalpophilus" SASP as well as for two B . cereus SASP were cloned, their nucleotide sequences were determined, and the amino acid sequences of the SASP coded for were compared . Strikingly, all of the amino acid residues previously found to be conserved in this group of SASP both within and between two other Bacillus species (B . megaterium and B . subtilis) were also conserved in the SASP coded for by the B . cereus genes as well as those coded for by the genes from the more distantly related organisms B . stearothermophilus and "T . thalpophilus." This finding strongly suggests that there is significant selective pressure to conserve SASP primary sequence and thus that these proteins serve some function other than simply amino acid storage. Int J Radiat Biol Relat Stud Phys Chem Med, 1986 Jul, 50(1), 31 - 4 Radiation sensitization of Bacillus megaterium spores by trans-dichlorodiammineplatinum(II); Powers EL et al.; The radiation sensitivity of spores of Bacillus megaterium is markedly increased by transplatin in both N2 and O2, and in a manner suggesting the involvement of hydroxyl radicals in both instances, thus differing mechanistically from its isomer cisplatin. J Virol, 1986 Jul, 59(1), 103 - 11 Bacteriocinlike killing action of a temperate bacteriophage phiBA1 of Bacillus aneurinolyticus; Ito S et al.; A new temperate phage, phiBA1, was isolated from Bacillus aneurinolyticus, phiBA1 had an icosahedral head with a diameter of about 70 nm and a tail about 20 nm long and contained a circularly permuted, linear duplex DNA of about 38 x 106 daltons . This phage showed two activities: bacteriocin-like killing activity against five strains of B . aneurinolyticus and normal temperate phage activity against three other strains . phiBA1 killed sensitive cells by a single-hit process . After adsorption of phiBA1 to cells sensitive to killing, the content of intracellular ATP increased for the first 5 min and then gradually decreased . Phage DNA injected into the cell immediately after infection was degraded rapidly . Killing was also caused by heavily UV-irradiated phiBA1 . Killing-resistant mutants showed normal adsorption of phiBA1 and normal injection of the DNA with its instantaneous restriction . Our results indicate that the killing action of phiBA1 is different from the phenomenon of abortive infection and suggest that the killing might be caused by a proteinaceous component of phiBA1. J Cell Sci, 1986 Jul, 83, 89 - 101 Characterization and partial purification of a plasma membrane receptor for Bacillus thuringiensis var . kurstaki lepidopteran-specific delta-endotoxin; Knowles BH et al.; The lepidopteran-specific P1 delta-endotoxin of Bacillus thuringiensis var . kurstaki HD-1 was activated in vitro using insect gut proteases and found to be highly specific for the lepidopteran cell line Choristoneura fumiferana CF1 among a wide range of lepidopteran and dipteran cell lines tested . The toxicity of P1 against CF1 cells is inhibited by N-acetylgalactosamine (GalNAc), and the lectins soybean agglutinin (SBA) and wheat-germ agglutinin . Protein blotting was used to identify a glycoprotein of 146 X 10(3) Mr in the plasma membrane of CF1 cells, capable of binding both the toxin and SBA, which is specific for GalNAc . This glycoprotein was labelled using galactose oxidase and sodium boro-{3H}hydride and solubilized in Triton X-100 before partial purification by affinity chromatography on SBA-agarose . We propose that this glycoprotein is a good candidate for the cellular receptor of the lepidopteran-specific P1 delta-endotoxin of B . thuringiensis var . kurstaki HD-1. Mol Gen Genet, 1986 Jul, 204(1), 52 - 7 Identification of Tn4430, a transposon of Bacillus thuringiensis functional in Escherichia coli; Lereclus D et al.; The mobile genetic element Tn4430, originating from the gram-positive bacterium, Bacillus thuringiensis, and previously described as the Th-sequence, is the first transposon isolated from the genus Bacillus . In the present work a gene (APH-III) conferring resistance to kanamycin was inserted into this 4.2 kb transposon . Transposition experiments showed that Tn4430 omega APH-III could transpose in the gram-negative host Escherichia coli when its insertion functions were supplied by an intact copy of Tn4430 . By transposing Tn4430 omega APH-III directly onto pBR322, it was possible to determine the nucleotide sequence of the terminal inverted repeats of Tn4430 and of the target DNA site . Identical 38 bp in inverted orientation are situated at each end of the transposon and there is a direct duplication of 5 bp at the insertion site . Thus, it is clear that Tn4430 is closely related to the transposons belonging to the Tn3 family (class II elements). J Bacteriol, 1986 Jul, 167(1), 30 - 4 Manganese reduction by a marine Bacillus species; de Vrind JP et al.; Mature dormant spores of marine Bacillus sp . strain SG1 catalyze the oxidation of Mn(II) to MnO2 . We report that vegetative cells of the same strain reduced MnO2 under low-oxygen conditions . The rate of reduction was a function of cell concentration . The process had a pH optimum of 7.5 to 8.0 and was inhibited by HgCl2, by preheating of the cells at 80 degrees C for 5 min, by antimycin A, and by N-heptyl-hydroxy-quinoline-N-oxide . At a nonlimiting O2 concentration, little MnO2 reduction was observed . Under these conditions, the process could be induced by the addition of NaN3 . Spectrophotometric analysis of the Bacillus cells indicated the presence of type b and c cytochromes . Both types can be oxidized in situ by addition of MnO2 to the cells. J Bacteriol, 1986 Jul, 167(1), 148 - 52 Predominance of gluconate formation from glucose during germination of Bacillus megaterium QM B1551 spores; Otani M et al.; Metabolic pathways of glucose during germination of Bacillus megaterium QM B1551 spores were studied by using specifically labeled glucose and gluconate . The Embden-Meyerhof pathway, the pentose cycle, and the direct oxidation route of glucose to gluconate (the gluconate pathway) were all operative at this stage; among those, gluconate accumulation was most predominant, especially in the early stage . Potassium fluoride, an enolase inhibitor, abolished the catabolism by the Embden-Meyerhof pathway totally without affecting gluconate accumulation . Under these conditions glucose was exclusively oxidized to gluconate . Gluconate thus accumulated could be metabolized further via phosphorylation by gluconate kinase . Remarkable gluconate accumulation was also demonstrated in several other spores requiring alanine as an effective germinant . NADH formed by the direct glucose oxidation may serve as a initial ATP source to phosphorylate glucose in germinating spores. Immunobiology, 1986 Jul, 171(4-5), 366 - 80 Autoreactive T cell clones from mice infected with Mycobacterium bovis, strain Bacillus Calmette-Guérin (BCG) . I . Phenotype, specificity and in vitro function; Muller I et al.; Mice were infected with the intracellular microorganism, Mycobacterium bovis BCG, and draining lymph node cells were collected . A T cell line was established which was cultured in the presence of syngeneic accessory cells (AC) and killed BCG . Stimulation of this line depended on syngeneic accessory cells and did not require BCG as a source of antigen, indicating that it was autoreactive . T cell clones derived from this line had the L3T4 helper/inducer phenotype and reacted with self-Ia on syngeneic macrophages or B cell blasts . Cloned T cells were also stimulated by syngeneic accessory cells pretreated with the lysosomotropic agent chloroquine and by H-2 compatible, background gene disparate, accessory cells, suggesting that they were specific for self-Ia . After in vitro stimulation, the T cell clones secreted interleukin 2 (IL 2) and interferon-gamma (IFN-gamma), helped B cells in antibody production and activated macrophages for secretion of reactive oxygen metabolites. Ann Inst Pasteur Microbiol, 1986 Jul-Aug, 137B(1), 101 - 11 Recycling of Bacillus sphaericus 2362 in mosquito larvae: a laboratory study; Charles JF et al.; After ingestion by Culex pipiens and Anopheles stephensi fourth instar larvae, spores of Bacillus sphaericus strain 2362 rapidly germinated inside live mosquito midgut . Bacterial counts and electron microscopic observations on intoxicated larvae revealed that the number of viable spores rapidly decreased during the first 12 h, with a minimum between 12 and 24 h . In cadavers, the number of heat-resistant spores quickly increased between the first and second day post-feeding . After one week, the number of spores inside dead larvae reached approximately 20 times the number of ingested spores for both mosquito species (ca . 4 X 10(5) spores/larva) . Ultrathin sections of recycled spores showed the presence of a crystalline inclusion identical to that initially present in spores before ingestion . Bioassay on C . pipiens fourth instar larvae showed a similar toxicity between in vivo recycled spores (LC50 = 1.1 +/- 0.3 X 10(5) spores/ml after 24-h exposure) and culture-medium-grown spores of B . sphaericus strain 2362 (LC50 = 1.7 +/- 0.4 X 10(5) spores/ml). Biochem Biophys Res Commun, 1986 Jun 13, 137(2), 748 - 51 Biochemical and immunological characterization of the cloned crystal toxin of Bacillus thuringiensis var . israelensis; Sekar V; The protein components of the cloned crystal toxin of Bacillus thuringiensis var . israelensis were separated by polyacrylamide gel electrophoresis under denaturing conditions . Using an antiserum to the solubilized B . thuringiensis var . israelensis crystal protein as a probe, immunological homology between the crystal protein components of B . thuringiensis var . israelensis and those of the recombinant B . megaterium strain VB131 was tested . The results from this study indicate that the crystal inclusion of the recombinant strain contains only the 130 kilodalton protein and not the 68 or the 28 kilodalton proteins of the crystal toxin of B . thuringiensis var . israelensis and that the 130 kilodalton protein is primarily responsible for the mosquitocidal activity of this organism. J Biol Chem, 1986 Jun 5, 261(16), 7160 - 9 Characterization of a catalytically self-sufficient 119,000-dalton cytochrome P-450 monooxygenase induced by barbiturates in Bacillus megaterium; Narhi LO et al.; A unique cytochrome P-450-dependent fatty acid monooxygenase from Bacillus megaterium ATCC 14581 is strongly induced by phenobarbital (Narhi, L . O., and Fulco, A . J . (1982) J . Biol . Chem . 257, 2147-2150) and many other barbiturates (Kim, B.-H., and Fulco, A . J . (1983) Biochem . Biophys . Res . Commun . 116, 843-850) . This monooxygenase has now been purified to homogeneity from pentobarbital-induced bacteria as a single polypeptide with a molecular weight of 119,000 +/- 5,000 daltons . In the presence of NADPH and O2, it can catalyze the oxygenation of long chain fatty acids without the aid of any other protein . The enzyme has a catalytic center activity of 4,600 nmol of fatty acid oxygenated per nmol of P-450 (the highest activity yet reported for a P-450-dependent monooxygenase) and also functions as a highly active cytochrome c reductase in the presence of NADPH . The purified holoenzyme is a soluble protein containing 40 mol % hydrophobic amino acid residues and 1 mol each of FAD and FMN/mol of heme . It is isolated and purified in the low spin form but is converted to the high spin form in the presence of long chain fatty acids . The enzyme, which catalyzes the omega-2 hydroxylation of saturated fatty acids and the hydroxylation and epoxidation of unsaturated fatty acids has its highest affinity (Km = 2 +/- 1 microM) for the C15 and C16 chain lengths. Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 141 - 51 Pronephric leucocytes of Cyprinus carpio: isolation, separation and characterization; Bayne CJ; Since the teleost pronephros is an important source of diverse immunocytes, suspensions of pronephric cells from young adult carp have been characterized . In freshly prepared suspensions, adherent, spreading cells (macrophages?) constituted less than 3% of the total population . Granulocytes and lymphocytes were co-dominant (less than 80%) leucocyte types . Continuous Percoll density gradient centrifugation yielded discrete subpopulations with these rho values and cytological characteristics: Fraction I & II rho = 1.055-1.070 thrombocytes, monocytes, macrophages, and lymphocytes . Fraction III rho = 1.080-1.090 granulocytes, type 1 . Fraction IV rho = 1.105-1.110 erythrocytes and granulocytes, type 2 . Fraction V rho = 1.118-1.125 granulocytes, type 3 . Fraction VI rho = 1.140-1.150 granulocytes, type 4 . Granulocyte motility increased markedly over the first 24 hr in vitro, and was enhanced by components washed from intact yeast . The subtypes of granulocytes were distinguishable by not only the rho values, but also on the basis of cell size, ultra-structure of the granules, and their histochemical and phagocytic characteristics . After simultaneous in vivo injection of Bacillus megaterium (Gram + ve), Aeromonas hydrophila (Gram - ve) and Saccharomyces cerevisiae (yeast), individual pronephric leucocytes were found capable of phagocytosing all three types of particle . Granulocytes which had phagocytosed B . megaterium were slower than macrophages in their ability to kill the bacteria . Encounter with B . megaterium or S . cerevisiae in vitro elicited a clumping reaction which involved mostly the larger leucocytes {granulocytes} . Both adherent cells and non-adherent cells were phagocytic in vitro. Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 127 - 40 Ultrastructural observations on peritoneal exudate cells from the striped bass; Bodammer JE; An ultrastructural study was conducted of the principal cells recovered from the peritoneal exudate of striped bass (Morone saxatilis) injected with killed Bacillus cereus . The most frequently observed cells were identified as macrophages, eosinophils, and neutrophils on the basis of comparing their structure with that reported for other fish species and higher vertebrates . All three of the types of cells were phagocytic; however, the numbers of bacteria engulfed were limited . Like other studies of this type in fish, the peritoneum of the striped bass proved to be a good source of cells for morphological investigations of phagocytosis. Arch Microbiol, 1986 Jun, 145(1), 51 - 5 Nickel-content of urease from Bacillus pasteurii; Christians S et al.; Urease from Bacillus pasteurii DSM 33 was purified 34-fold to a maximum specific activity of 996.5 mumol urea min-1 mg-1 at 30 degrees C . Homogeneity was demonstrated by isoelectric focussing which showed a single protein zone corresponding to a pI of about 4.6 . The native enzyme was demonstrated to have a molecular mass of 230,000 and to consist of identical subunits of 65,500, as measured by SDS electrophoresis . Radioactive 63Ni-nickel co-chromatographed with urease through gel filtration, ion-exchange, and affinity chromatography . Measuring specific radioactivity, the nickel content was found to be 1.00 (+/- 0.1) g-atom Ni per mol of subunit, and 0.82 g-atom Ni per mol as measured by atomic absorption spectrometry . This indicates that 1 atom of nickel is present in each of four subunits of the enzyme. J Dairy Sci, 1986 Jun, 69(6), 1510 - 7 Detection of penicillin, cephapirin, and cloxacillin in commingled raw milk by the Spot test; Ryan JJ et al.; The objective of this study was to compare Spot Test results with the results of the Bacillus stearothermophilus disc assay . Commingled raw milk samples were subdivided and spiked with penicillin, cephapirin, or cloxacillin . All subsamples, including unspiked subsamples, were analyzed by the Spot Test (3 or 9 replicates) and disc assay (9 replicates) . Mean zone diameter for every subsample was calculated; subsamples having a mean zone diameter greater than or equal to 16 mm were considered positive . At penicillin concentrations less than or equal to 3.7 ng/ml, agreement between the Spot Test and disc assay was 83.4%, and false positive and negative percentages were 14.4 and 2.2%, respectively . Above 3.7 ng/ml, agreement was 100% . At cephapirin concentrations less than or equal to 12 ng/ml, agreement between the two tests was 67.9%, and false positive and negative percentages were 28.8 and 3.3%, respectively . Above 15 ng/ml, agreement was 99.3% with .7% false negatives . At cloxacillin concentrations less than or equal to 50 mg/ml agreement between the two tests was 54.5 with 45.5% false positives . At cloxacillin concentrations greater than or equal to 62.5 ng/ml, agreement between the two tests was 87.2% and false positive and negative percentages were 12.6 and .2%, respectively . In a field trial consisting of 823 bulk samples, Spot Test and disc assay agreement was 100% . The Spot Test is a rapid and reliable method for detecting penicillin, cephapirin, and cloxacillin residues in raw milk at concentrations that will produce a 16-mm zone. Am J Public Health, 1986 Jun, 76(6), 643 - 6 Tuberculin reactivity in United States and foreign-born Latinos: results of a community-based screening program; Perez-Stable EJ et al.; Because of the concern that we were underestimating the prevalence of tuberculosis within the Latino community in San Francisco, we undertook a community-based screening program directed largely towards recent immigrants . Of 1,871 intermediate-strength (5 TU) tuberculin tests applied and read, 37 per cent of the reactions were greater than or equal to 10 mm . Significant reactions were found in 53 per cent of foreign-born persons compared to 7 per cent of those born in the United States . Persons older than 20 years of age were more likely to have significant reactions compared to younger Latinos . Among the foreign-born, the frequency of significant reactions was not influenced by the length of stay in the US or a history of BCG (bacille Calmette-Guerain) vaccination . Two foreign-born children were found to have current tuberculosis . The prevalence of tuberculin reactors among US-born Latino children was 3 per cent, which suggests that undetected transmission of tuberculosis may be occurring . We conclude that Latino immigrants should be systematically screened for tuberculosis. Antibiot Med Biotekhnol, 1986 Jun, 31(6), 405 - 8 {Disordered bacitracin synthesis and sporulation in mutant Bacillus licheniformis with decreased proteolytic activity}; Lukin AA et al.; The vegetative cells of B . licheniformis, producing bacitracin, a feed grade peptide antibiotic were exposed to nitroso guanidine and six mutants with lowered activity of serine exoprotease in the culture fluid were isolated . It was found that synthesis of bacitracin and sporulation of the mutants were impaired . The cause of the impairment is not known . The results are discussed from two viewpoints: serine exoprotease is responsible for positive regulation of bacitracin biosynthesis and sporulation at the account of forming an amino acid pool in the medium, simultaneous changing of the three features in the mutants may be due either to a single pleiotropic mutation or to multiple mutations independently induced by nitroso guanidine. Int J Lepr Other Mycobact Dis, 1986 Jun, 54(2), 256 - 67 Antibodies to phenolic glycolipid-1 and to whole Mycobacterium leprae in leprosy patients: evolution during therapy; Bach MA et al.; Sera from 92 patients were tested by the ELISA method for the presence of IgM antibodies to phenolic glycolipid-1 (PGL-1) of Mycobacterium leprae, and of both IgM and IgG antibodies to the whole M . leprae bacillus . All untreated lepromatous patients exhibited high antibody levels in all three assays . A sharp decline of IgM antibodies to PGL-1 and whole M . leprae was observed during the first two years of therapy, while IgG antibodies to whole M . leprae showed a progressive decrease only over a number of years . Low titers of IgM antibodies to PGL-1 and IgG antibodies to whole M . leprae could be detected in about 50% and 75% of patients, respectively, after more than ten years of treatment, with only 15% showing persisting IgM antibodies to the whole bacillus . Antibody levels as measured by the three assays used were correlated with the bacterial index in patients treated for less than four years . In patients treated longer than four years, only IgM antibodies, whether directed to PGL-1 or to whole M . leprae, remained correlated to the bacillary load . Tuberculoid patients exhibited a different antibody pattern, showing a lower frequency (and lower levels) of antibodies of PGL-1 and of IgG antibodies to whole M . leprae than lepromatous patients, and no detectable IgM antibodies to the whole bacillus . IgG antibodies to whole M . leprae were more frequently noted than antibodies to PGL-1, the latter declining more rapidly during therapy. Poult Sci, 1986 Jun, 65(6), 1217 - 9 The effect of Bacillus natto on the T and B lymphocytes from spleens of feeding chickens; Inooka S et al.; The effect of Bacillus natto on T and B lymphocytes in the chicken spleen was investigated . Bacillus natto is isolated from fermented food and is used in Japan in a preparation that enhances growth of farm animals . When chickens were fed 10(7) colony-forming units of B . natto (in spores) per gram of feed from hatching to 27 days of age there was an increase in the percentage of T and B lymphocytes in the spleens compared with non-B . natto-fed controls . The findings suggested that dietary Bacillus natto had an effect on cellular immune responses. Vet Immunol Immunopathol, 1986 Jun, 12(1-4), 251 - 62 Paradoxical effects of cadmium exposure on antibacterial antibody responses in two fish species: inhibition in cunners (Tautogolabrus adspersus) and enhancement in striped bass (Morone saxatilis); Robohm RA; Previous work in a marine fish, the cunner (Tautogolabrus adspersus), showed that endocytosis of bacteria by cells in the liver and spleen was affected by 96-h exposure of the fish to cadmium at a concentration of 12 micrograms/ml; however, antibody response to sheep erythrocytes was not affected . Since the latter finding was questionable because of short immunization times, and data from more than a single fish species were desirable, both the cunner and an anadromous fish, the striped bass (Morone saxatilis), were examined for antibody responses against the bacterium Bacillus cereus in Freund's complete adjuvant during a 6- to 8-wk time period . Exposure to 12 micrograms/ml cadmium caused significant inhibition of serum antibody titers (P less than 0.007) in cunners . Paradoxically, antibody response in striped bass exposed to 10 micrograms/ml cadmium was enhanced sixfold . This enhancement was weaker but still evident when the antigen was injected 48 d after cadmium exposure . Peritoneal exudate cells from cadmium-exposed striped bass also showed more active migration through microporous filters than cells from non-exposed fish . Since the 96-h cadmium LC50 was 26 micrograms/ml for cunners and 20 micrograms/ml for striped bass, the differences in antibody response could not be explained on the basis of differences in cadmium toxicity . Although geometric mean liver cadmium levels after exposure at 15 degrees C were higher in cunners (163.8 +/- 1.2 micrograms/gm) than in striped bass (64.4 +/- 1.2 micrograms/g), cunners exposed to cadmium at 2 degrees C had lower cadmium levels (46.4 +/- 1.2 micrograms/g) which were still effective in inhibiting antibody production at 8 degrees C . On the other hand, striped bass not exposed to cadmium showed a strong, exponential rise in serum antibody when the temperature at immunization, 14 degrees C, was reduced to 9 degrees C; whereas cunners held in the same tanks exhibited a weaker, biphasic serum antibody response . Regardless of the cause (innate differences in cellular response, or better stress adaptability in an anadromous fish), the data show that chemical-stress effects on the immune system of one fish species cannot be extrapolated to another species. Arch Microbiol, 1986 Jun, 145(1), 97 - 103 Turnover of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae: differences between in vivo and in vitro degradation; Chopra AK et al.; Degradation of abnormal proteins in Bacillus megaterium and Saccharomyces cerevisiae in vivo was compared with that in cell-free extracts . Protein degradation in vivo, when the cells were labelled with 14C-leucine during growth in the presence of ethionine, was affected by the concentration of the analogue used . Proteins synthesized in the presence of 0.2-1 mM ethionine were degraded most rapidly in both organisms . The proteolytic enzyme system of yeast degraded the analogue-containing proteins in vitro faster than the normal proteins . This holds also for proteins synthesized in the presence of 5 mM ethionine, whose degradation in vivo was impaired . The proteolytic system of B . megaterium, on the other hand, was unable in vitro to differentiate between normal and abnormal proteins . Denatured proteins underwent preferential degradation over normal and ethionine-containing proteins. Agents Actions, 1986 Jun, 18(3-4), 413 - 20 Studies on type II collagen induced arthritis in mice; Paska W et al.; A consistent and reproducible polyarthritis was induced in mice by immunizing them with type II collagen in Complete Freunds adjuvant (CFA) and Bacillus Calmette-Guerin (BCG) vaccine . Several inbred strains of mice were investigated for the ability to develop collagen induced arthritis (CIA) . DBA/1 mice (H-2q) produced the highest incidence and the most severe arthritis of all the strains examined . Viable BCG vaccine was essential for the induction of a reproducible disease in this strain . The effects of some anti-inflammatory and anti-rheumatic compounds were examined on the developing and established lesions of CIA . These effects were determined by assessing the paw inflammation using a subjective scoring system and measuring foot weight . Furthermore, levels of serum amyloid P component (SAP) were also determined . Benoxaprofen, cyclophosphamide, indomethacin and prednisolone inhibited the paw inflammation in the developing disease whilst the anti-rheumatic compounds auranofin and D-penicillamine exacerbate the paw inflammation . Cyclophosphamide and prednisolone inhibited the established lesions but only prednisolone prevented the development of further lesions in the established disease . The SAP levels in the prednisolone treated group were also reduced . Auranofin treatment exacerbated the inflammation of both the established and the developing lesions in the same animal . D-penicillamine was inactive in the established disease. J Appl Bacteriol, 1986 Jun, 60(6), 513 - 6 Survival and growth of Bacillus cereus in bread; Kaur P; Bread doughs were artificially inoculated with spores of six Bacillus cereus strains at different inoculum levels and counts of survivors in bread determined during storage at 27.5 degrees C . No B . cereus were isolated from the centre crumb of 400 g loaves when the dough contained less than 10(4) spores/g whereas with 800 g loaves survival occurred with doughs containing 5.0 X 10(3) spores/g . With all strains there was a period of at least 24 h before multiplication took place in the bread . The inclusion in dough of 0.2% of calcium propionate, based on flour, effectively delayed germination and subsequent multiplication of B . cereus spores . It is concluded that the risk of food poisoning due to the presence of B . cereus in bread is minimal. J Antibiot (Tokyo), 1986 Jun, 39(6), 755 - 61 Plipastatins: new inhibitors of phospholipase A2, produced by Bacillus cereus BMG302-fF67 . III . Structural elucidation of plipastatins; Nishikiori T et al.; Plipastatins are new inhibitors of phospholipase A2 produced by Bacillus cereus BMG302-fF67 . Structures of the plipastatins have been determined by UV, mass and NMR spectrometries and chemical degradation . The carboxyl group of the C-terminal L-isoleucine of plipastatinic acid has been shown to form a lactone linkage with the hydroxyl group of L-tyrosine . The total structure of plipastatins has thus been established. J Antibiot (Tokyo), 1986 Jun, 39(6), 745 - 54 Plipastatins: new inhibitors of phospholipase A2, produced by Bacillus cereus BMG302-fF67 . II . Structure of fatty acid residue and amino acid sequence; Nishikiori T et al.; Plipastatins, new inhibitors of phospholipase A2, were produced by a strain classified as Bacillus cereus BMG302-fF67 . The plipastatins are a family of acylated decapeptides which differ from each other by amino acid composition and the nature of the fatty acid side chain . The fatty acids have been shown to be 3(R)-hydroxyhexadecanoic acid (n-C16h3) and 14(S)-methyl-3(R)-hydroxyhexadecanoic acid (a-C16h3) by mass, NMR and optical rotation . Amino acid sequence analysis by secondary ion mass spectrometry and additional physico-chemical evidence indicate that the structures of plipastatinic acids, the lactone-opened peptides are as follows: beta-Hydroxy fatty acid----L-Glu----D-Orn----L-Tyr----D-allo-Thr----L-Glu----D-Ala(Val)---- L-Pro----L-Gln----D-Tyr----L-Ile X OH. J Antibiot (Tokyo), 1986 Jun, 39(6), 737 - 44 Plipastatins: new inhibitors of phospholipase A2, produced by Bacillus cereus BMG302-fF67 . I . Taxonomy, production, isolation and preliminary characterization; Umezawa H et al.; Plipastatins have been isolated as part of a program designed to find inhibitors of porcine pancreatic phospholipase A2 . They were purified from fermentation broth of Bacillus cereus BMG302-fF67 and finally separated into four fractions by reverse phase HPLC . The respective fractions were designated as plipastatins A1, A2, B1 and B2 . Plipastatins also inhibited phospholipases C and D. Can J Microbiol, 1986 Jun, 32(6), 522 - 4 Inhibition of L-lysine uptake by alpha-methyl lysine in Escherichia coli and Bacillus sphaericus; Regenstreif CA et al.; alpha-Methyl lysine was investigated as a potential inhibitor of lysine transport in Escherichia coli and Bacillus sphaericus . At equimolar concentrations, no inhibition was observed in either organism, but at 10X and 100X the lysine concentration, alpha-methyl lysine caused a 20-50% reduction in the initial rate of lysine uptake in both bacteria . A similar inhibitory effect was observed with epsilon-N-methyl lysine on lysine uptake in B . sphaericus, but not in E . coli . alpha-Methyl lysine had a reduced effect on ornithine uptake and no effect on arginine transport in either bacterium. Proc Natl Acad Sci U S A, 1986 Jun, 83(11), 3977 - 81 Control of hemopoiesis in mice by sensitized L3T4+ Lyt2-lymphocytes during infection with bacillus Calmette-Guérin; Marchal G et al.; When injected intravenously with bacillus Calmette-Guerin (BCG; 10(7) viable units), C57BL/6 mice rapidly develop a transient anemia associated with an increased number of granulocytes and monocytes, whereas C3H/He mice do not . Because these two features are lacking in C57BL/6 nude mice we postulated that T lymphocytes can regulate hemopoiesis during infection . To assess further the role in hemopoiesis of T lymphocytes present in bone marrow of C57BL/6 and C3H/He mice, the frequency of BCG-specific T lymphocytes and their surface marker phenotype were determined by limiting dilution analysis and use of monoclonal antibodies . The number of BCG-specific T lymphocytes was estimated to be 50- to 100-fold higher in bone marrow of C57BL/6 than in that of C3H/He mice . Although L3T4+ Lyt2-and L3T4- Lyt2+ BCG-specific T lymphocytes were generated in mice of both strains, in C57BL/6 mice L3T4+ cells were induced preferentially from day 1 through day 5 after infection in correlation with hemopoietic changes . The relation between T-cell immune response and hemopoietic changes was substantiated by results obtained after in vivo treatment with monoclonal antibodies . Selective depletion of L3T4+ T cells by in vivo injection of anti-L3T4 monoclonal antibodies (GK 1-5) inhibited the development of the anemia and the related increased production of phagocytes in C57BL/6 mice receiving BCG. J Bacteriol, 1986 Jun, 166(3), 722 - 7 Structure of a beta-galactosidase gene of Bacillus stearothermophilus; Hirata H et al.; The nucleotide sequence of the bgaB gene, which encodes the thermostable beta-galactosidase I of Bacillus stearothermophilus, and its flanking region was determined . A 2,016-base-pair open reading frame observed was concluded to be for beta-galactosidase I (Mr 78,051) from observations that the amino acid composition of the enzyme and the sequence of 14 amino acids from the amino-terminus of the enzyme coincided with those deduced from this open frame . A 107-base-pair HaeIII-AluI fragment just upstream of the estimated Shine-Dalgarno sequence of the bgaB gene had promoter activity toward cat-86 (chloramphenicol acetyltransferase gene) and produced the enzyme at a level equivalent to 7% of the total cellular protein of B . subtilis . From the base sequence of this DNA region and the transcriptional start site determined by S1 nuclease mapping, the -35 and -10 sequences are estimated to be TTGACA and TAATTT, respectively, which are similar to the consensus sequence of B . subtilis sigma 43 RNA polymerase. Lab Anim Sci, 1986 Jun, 36(3), 271 - 6 Diagnosis of murine infections in relation to test methods employed; Kraft V et al.; Comparative investigations of Sendai virus, pneumonia virus of mice (PVM), mouse encephalomyelitis virus (mouse polio), minute virus of mice (MVM), and reovirus type 3 (Reo 3) infected murine colonies revealed a 30% higher incidence of positive sera when enzyme-linked immunosorbent assay (ELISA) was employed instead of hemagglutination inhibition (HI) tests . Equivalent sensitivity as in the ELISA was obtained when the same sera were investigated by indirect immunofluorescence (IF) tests . The virus purification techniques described resulted in highly suitable antigens for all indirect ELISA established . Since IIF requires no purified antigens, this test is recommended as an alternative to ELISA as well as to HI and complement fixation (CF) tests for laboratories lacking the necessary equipment for high speed centrifugation . A high incidence of false positive HI reactions was found particularly in Reo 3 routine serology . An updated survey of seromonitoring showed that European murine colonies appeared to be infected far less with Reo 3 if ELISA or IIF tests were employed . During 1982-1984, only 13% of the mouse colonies screened possessed Reo 3 positive sera whereas no natural Reo 3 infection was found in rat colonies . Mouse hepatitis virus (MHV) and the coronaviruses of rats exhibited the highest incidence in murine colonies . A total of 60% of mouse and 41% of rat colonies were found to be infected by these viruses . In comparison with earlier serological surveys, the relative incidence of other murine infections was similar . Antibodies against Bacillus piliformis (Tyzzer's disease) were detected by the IIF test in 41% of the rat colonies screened. J Bacteriol, 1986 Jun, 166(3), 728 - 32 Excretion of the penicillinase of an alkalophilic Bacillus sp . through the Escherichia coli outer membrane is caused by insertional activation of the kil gene in plasmid pMB9; Kobayashi T et al.; Most of the cloned penicillinase from alkalophilic Bacillus sp . strain 170 and alkaline phosphatase were released into the culture medium by Escherichia coli strains bearing plasmid pEAP1 or pEAP2 (T . Kudo, C . Kato, and K . Horikoshi, J . Bacteriol . 156:949-951, 1983) . We analyzed the basis for excretion of periplasmic enzymes in the cells bearing these plasmids . Several experiments such as subcloning, insertion of a chloramphenicol acetyltransferase cartridge, and DNA sequencing were done . A dormant kil gene in plasmid pMB9 was expressed by a promoter of the inserted DNA fragment of alkalophilic Bacillus sp . strain 170, and as a result, the outer membrane of E . coli became permeable, allowing the proteins to be excreted without cell lysis. Am J Vet Res, 1986 Jun, 47(6), 1363 - 5 Drug residues in milk after intrauterine injection of oxytetracycline, lincomycin-spectinomycin, and povidone-iodine in cows with metritis; Kaneene JB et al.; A study was conducted to document the maximum retention times of antimicrobial residues in milk after their use in intrauterine treatment of metritis in lactating cows and to evaluate several risk factors hypothesized to influence the retention time of these drugs . Oxytetracycline (3 g), lincomycin-spectinomycin (2 g of one-third lincomycin and two-thirds spectinomycin), or povidone-iodine (6 g) were given to cows with metritis by intrauterine route . The Bacillus stearothermophilus var calidolactis disk assay was performed on each milk sample . Of the 61 cows treated with oxytetracycline, 30 had residues in their postinjection milk for variable periods (range, 12.5 to 44.0 hours; mean, 26.6 +/- 10.3) . Of the 47 cows treated with lincomycin-spectinomycin, 17 had residues in their postinjection milk for various periods (range, 14.5 to 24 hours; mean, 19.5 +/- 8.9) . Povidone-iodine was not detected in milk . Because a high number of cows (n = 61) were treated with oxytetracycline, only data from these cows were used in testing the influence of 3 factors (severity of metritis, time after parturition when cows with metritis were treated, and parity) on maximum retention of the drug in milk . Severity of metritis did not have a significant influence (P greater than or equal to 0.1) on the maximum retention time of the drug . The retention time decreased linearly with the increase of time after parturition when the cow with metritis was treated . First lactation cows had a significantly (P less than or equal to 0.01) shorter retention time than did older cows. J Bacteriol, 1986 Jun, 166(3), 769 - 78 Transcriptional analysis of beta-lactamase regulation in Bacillus licheniformis; Salerno AJ et al.; The expression of the blaP gene for the beta-lactamase of Bacillus licheniformis was examined by transcriptional analyses . Radiolabeled probes containing the blaP gene or various regions 3' or 5' to it were used to analyze RNA samples prepared from induced and uninduced cultures of wild-type and mutant B . licheniformis strains . The level of blaP mRNA was low in uninduced wild-type cells . At 37 degrees C, blaP mRNA levels reached a maximum 1 h after induction while rising up to 180-fold and then declined, but remained severalfold above the uninduced level for several hours . The rate of beta-lactamase synthesis was roughly proportional to the levels of blaP mRNA in both wild-type and mutant strains, indicating that regulation of beta-lactamase formation occurs primarily at the level of transcription . Turnover of blaP mRNA in the presence of rifampin was rapid, giving a blaP mRNA half-life of about 2 min . Yet, high levels of blaP mRNA were maintained for at least 1 h after removal of free inducer . Three blaP mRNAs of 1.2, 2.9, and 3.4 kilobases were produced from the blaP promoter . The most abundant made up about 97% of all blaP transcripts and was also the smallest, ending at a transcriptional terminator located about 60 bases 3' to the blaP structural gene . Variables such as incubation temperature, cytotoxicity of inducer, and type of strain had essentially no effect on the ratio of large blaP mRNA to total blaP mRNA . The 2.9- and 3.4-kilobase blaP mRNAs identify potential locations of genetically linked regulators of beta-lactamase synthesis. J Immunol, 1986 Jun 1, 136(11), 4328 - 33 Mechanisms of tumor cell capture by activated macrophages: evidence for involvement of lymphocyte function-associated (LFA)-1 antigen; Strassmann G et al.; The lymphocyte function-associated (LFA)-1 molecule is expressed on certain populations of macrophages that have an augmented capacity to capture tumor cells . Accordingly, we analyzed the role of LFA-1 in the establishment of such cell-cell interactions . F(ab')2 fragments of the M17/4, anti-LFA-1 monoclonal antibody (MAb) inhibited the interaction between activated macrophages and tumor cells by up to 80% in a dose-dependent manner . The anti-LFA-1 MAb reduced (between 55 to 79%) the number of P815, LSTRA, or EL-4 tumor cells bound to trypsin-sensitive structures on bacillus Calmette Guerin activated macrophages . The inhibition appeared selective, because a F(ab')2 fragment of anti-Mac-1 did not inhibit such binding . Inhibition of tumor cell capture could be observed as soon as 15 min after the onset of the cell-cell interaction between activated macrophages and tumor cells . Optimal inhibition occurred when both tumor targets and macrophages were precoated with the MAb . Although P815, LSTRA, EL-4, and BW5147 tumor cells all expressed LFA-1, only the first three but not BW5147 cells were bound by activated macrophages . Furthermore, endotoxin-pulsed macrophages elicited by thioglycollate broth expressed the LFA-1 antigen but did not exhibit selective tumor cell capture . Finally, anti-LFA-1 inhibited the development of weak into strong binding . Taken together, the results suggest that LFA-1 molecules can participate in the interaction between activated macrophages and neoplastic cells. J Immunol, 1986 Jun 1, 136(11), 4255 - 63 M . leprae and PPD-triggered T cell lines in tuberculoid and lepromatous leprosy; Shankar P et al.; Proliferative responses of peripheral blood mononuclear cells (PBMC) to Mycobacterium leprae and bacillus Calmette Guerin-derived purified protein derivative (PPD) were studied in the presence or absence of interleukin 2 (IL 2) in high M . leprae responders (tuberculoid leprosy patients and healthy subjects) and low M . leprae responders (lepromatous leprosy patients) . High responders in most cases developed a strong proliferative response to both antigens in the absence of IL 2 . Additional IL 2 and restimulation with antigen plus autologous antigen-presenting cells (APC) allowed the derivation of antigen-specific T cell lines . The lines were assayed for proliferative responses to several mycobacterial antigens . Both PPD and M . leprae-triggered T cell lines exhibited a good proliferative response to either antigen and showed in addition a broad cross-reactivity with other mycobacteria, suggesting a preferential T cell response to epitopes shared by several mycobacterial species . Within the lepromatous group, 50% of the patients studied could mount a proliferative response to PPD antigen in the absence of IL 2, but none of them was able to do so with M . leprae antigen . The addition of IL 2 increased the number of positive responders to PPD in this group, and in some patients IL 2 was able to restore M . leprae reactivity as well, suggesting that IL 2 had overcome a suppressor mechanism . PPD and M . leprae-triggered T cell lines were obtained from these subjects (with IL 2 added from the beginning of the culture when required) . M . leprae lines exhibited variable and unstable pattern of specificity, most lines exhibiting, at least transiently, a cross-reactive response to other mycobacteria, but some displaying only M . leprae-specific response . In contrast, PPD lines from these subjects consistently exhibited a good response to PPD, a lesser response to various other mycobacteria and no response to M . leprae, a pattern differing from that obtained with PPD lines of high M . leprae responders . Co-cultures of irradiated lepromatous PPD triggered T cell lines with fresh autologous PBMC non-specifically reduced the proliferative response of the latter to PPD, as well as to unrelated antigens . A similar suppression was also observed when PPD lines from one of the tuberculoid patients were assayed . PPD and M . leprae T cell lines from both high and low responders initially exhibited the same CD4+ CD8- phenotype . In all cases, antigenic specificity declined and could not be maintained after 5 to 8 wk of continuous culture, a change associated with the progressive appearance of CD8+ and Leu8+ cells. Am J Med, 1986 May 30, 80(5C), 2 - 12 Concept of empiric therapy with antibiotic combinations . Indications and limits; Klastersky J; It appears that the use of antibiotic combinations, especially synergistic ones, is indicated for the management of gram-negative bacillary sepsis in granulocytopenic patients . Synergism is a valuable factor in increasing the serum bactericidal activity, which is highly likely to be important for a favorable outcome in these infections . The potential side effects of antimicrobial combinations should not deter clinicians from their use . The most frequently used combinations for gram-negative bacillary infections are those involving beta-lactams and aminoglycosides . Other potentially synergistic combinations exist as well; however, the clinical experience with these combinations is limited, and, as with double beta-lactam combinations, their potential for antagonism necessitates care when using them . Besides gram-negative bacillary sepsis in granulocytopenic patients, severe staphylococcal infections might represent an indication for the use of combination therapy, especially in patients with compromised mechanisms of defense against infection. Biochem Biophys Res Commun, 1986 May 29, 137(1), 80 - 6 Preparation and sequencing of the cloacin fragment of Streptomyces aureofaciens 16S RNA; Janda I et al.; A fast method for isolation of a 3'-terminal fragment of Streptomyces aureofaciens 16S RNA was developed . The procedure involves reaction of 70S ribosomes with cloacin DF13 and subsequent fractionation of the reaction mixture by polyacrylamide gel electrophoresis . The cloacin fragment was eluted from the gel and used directly for 3'-end labeling with cytidine-3',5'-{5'-32P}bisphosphate . The labeled RNA fragment was sequenced by the enzymatic method . It consists of 50 nucleotides and has the sequence 5'-GUCGUAACAAGGUAACCGUACCGGA-AGGUGCGGUUGGAUCACCUCCUUUCOH . The differences from the E . coli and Bacillus sequences and their possible influence on the rate and specificity of polypeptide synthesis are discussed. J Immunol, 1986 May 15, 136(10), 3864 - 71 Induction of protective immunity against Schistosoma mansoni by a nonliving vaccine . II . Response of mouse strains with selective immune defects; James SL et al.; The efficacy of a new vaccination procedure against Schistosoma mansoni, involving intradermal injection of nonliving antigen combined with the bacterial adjuvant Mycobacterium bovis strain bacillus Calmette Guerin, was tested in several strains of mice . Development of protection against subsequent infection was compared with in vivo skin test reactivity and in vitro humoral reactivity to soluble and surface-associated schistosome antigens . Significant levels of resistance and immune response were displayed by many inbred mouse strains, including C57BL/6J, C3H/HeN, and CBA/J, as well as outbred Swiss-Webster mice . However, no definite correlation was observed between the level of any particular immune response and the level of resistance to challenge S . mansoni infection . Development of protective immunity was also examined in mice with various immune defects, to determine whether these responses are relevant to resistance in this model . Animals with defective specific immediate hypersensitivity response due to deficiencies in IgE (SJL/J) or mast cell (W/Wv) production displayed strong resistance as a result of immunization . Likewise, mice bearing the lpsd (C3H/HeJ) or xid (CBA/N) mutations, affecting cellular or humoral response to certain thymus-independent antigens, developed significant levels of resistance after immunization . A/J mice, with defects in cellular recognition of bacterial endotoxin as well as deficiencies in natural killer cell activity and complement function, also showed significant protective immunity . Thus, these reactivities do not appear to be essential to the resistance against S . mansoni induced by the nonliving vaccine . Two nonresponder strains were identified, P and BALB/c . P mice were defective in specific delayed hypersensitivity response as well as resistance to infection . However, BALB/c mice showed no obvious immune deficiencies at the time of challenge . These results agreed with previous findings in mice immunized by exposure to radiation-attenuated cercariae with one exception; BALB/c mice were protected by vaccination with irradiated cercariae but not by the nonliving vaccine . Thus, further examination of immune response in mice identified in this study as high and low responder strains should allow characterization of critical immune resistance mechanisms induced by the nonliving vaccine, as well as immune mechanisms operating in common between these two models of resistance to S . mansoni. Biochem Biophys Res Commun, 1986 May 14, 136(3), 1142 - 7 Further evidence for the role of 26 kDa peptide as mosquito larvicidal principle of the crystalline delta endotoxin of Bacillus thuringiensis var Israelensis; Sriram R et al.; Separation of the toxic and non-toxic subunits of the crystals of B . thuringiensis var israelensis, in native gels has revealed that the toxic subunit contained predominantly 26 kDa peptide, while the non-toxic subunit was made up of 66 kDa and other larger molecular weight peptides . Tryptic digestion of the crystal proteins and their separation on a DEAE-cellulose column, resulted in the generation of a fraction with a 21 kDa peptide, exhibiting larvicidal and hemolytic activities and immunoreactive with the antiserum of 26 kDa peptide of the crystals . These results show that 26 kDa peptide is the active principle of the crystal. Biochemistry, 1986 May 6, 25(9), 2707 - 13 Study of the interaction between uncharged yeast tRNAPhe and elongation factor Tu from Bacillus stearothermophilis; Heerschap A et al.; Proton NMR studies are presented on the interaction of nonaminoacylated yeast tRNAPhe and elongation factor Tu X GTP from Bacillus stearothermophilis . From experiments in which transfer of magnetization is observed between proton spins of tRNA and the protein, it is concluded that complex formation takes place . Amino acid residues of the protein come into close contact with the base pair A5U68 and/or U52A62 of the acceptor T psi C limb of the tRNA molecule . From the line broadening of tRNA resonances, associated with complex formation, an association constant of 10(3)-10(4) M-1 is estimated . The NMR experiments do not monitor a significant conformational change of the tRNA molecule upon interaction with the protein . However, at times long after the onset of complex formation, spectral changes indicate that the upper part of the acceptor helix becomes distorted. Eur J Biochem, 1986 May 2, 156(3), 531 - 40 Specificity of Bacillus thuringiensis var . colmeri insecticidal delta-endotoxin is determined by differential proteolytic processing of the protoxin by larval gut proteases; Haider MZ et al.; The native crystal delta-endotoxin produced by Bacillus thuringiensis var . colmeri, serotype 21, is toxic to both lepidopteran (Pieris brassicae) and dipteran (Aedes aegypti) larvae . Solubilization of the crystal delta-endotoxin in alkaline reducing conditions and activation with trypsin and gut extracts from susceptible insects yielded a preparation whose toxicity could be assayed in vitro against a range of insect cell lines . After activation with Aedes aegypti gut extract the preparation was toxic to all of the mosquito cell lines but only one lepidopteran line (Spodoptera frugiperda), whereas an activated preparation produced by treatment with P . brassicae gut enzymes or trypsin was toxic only to lepidopteran cell lines . These in vitro results were paralleled by the results of in vivo bioassays . Gel electrophoretic analysis of the products of these different activation regimes suggested that a 130-kDa protoxin in the native crystal is converted to a 55-kDa lepidopteran-specific toxin by trypsin or P . brassicae enzymes and to a 52-kDa dipteran toxin by A . aegypti enzymes . Two-step activation of the 130-kDa protoxin