Genetika, 1983 Mar, 19(3), 406 - 15 {Simultaneous reversion for genes thy, dra and drm in Bacillus thuringiensis}; Grigor'eva TM et al.; The reversions of mutations in genes thy, dra (deoxyriboaldolase) and drm (phosphodeoxyribomutase) were studied in Bacillus thuringiensis var . galleriae 351 . In this strain, both dra and drm mutants are sensitive to the presence of thymidine (Tds) . In addition to the reversions for one of the properties (thymine requirement or sensitivity to thymidine), the simultaneous reversions for both properties (Thy + Tdr) were indicated . The frequency of Thy + Tdr revertants varied from 10(-4) to 10(-8), depending on both a selection method and the conditions of preliminary cultivation . The Thy + Tdr revertants had a rough (R) colony morphology in 100% of the cases, though the parental strains were of smooth (S) colony morphology . Selection for R variants, visually or by means of screening for Tg4 resistant or tetracycline resistant clones, resulted in detecting double Thy + Tdr revertants in 100% of the cases . The parental Thy - Tds phenotype restored after R leads to S conversion which occurred at the frequency 10(-7) to 10(-8) and might be detected by means of streptomycine selection. J Clin Lab Immunol, 1983 Mar, 10(3), 143 - 6 Bacteria-immune system interactions . IV . Effect of various treatments on the binding of bacteria to human lymphocytes; Shockley RK et al.; Pretreatment of lymphocytes with selected saccharides inhibited the binding of Bacillus globigii (B . globigii) to lymphocytes but had no effect on Escherichia coli 2 (E . coli 2) adherence . The binding of both bacteria was prevented by lymphocyte pretreatment with trypsin or pronase . Periodate treatment of lymphocytes inhibited the binding of E . coli 2 but not of B . globigii . This inhibition could be reversed by borohydrate treatment of lymphocytes . Trypsin, pronase or periodate treatment of bacteria had no effect on the binding to lymphocytes . The data obtained suggest that: (a) E . coli 2 and B . globigii have different mechanisms of binding to lymphocytes; and (b) the lymphocyte receptor for E . coli 2 is of a glycoprotein nature. Infect Control, 1983 Mar-Apr, 4(2), 81 - 9 A prospective study of infectious diseases following bone marrow transplantation: emergence of Aspergillus and Cytomegalovirus as the major causes of mortality; Peterson PK et al.; We performed a prospective study of infections following bone marrow transplantation in 50 patients treated for aplastic anemia or hematologic malignancy . Early, continuous prophylaxis with trimethoprim/sulfamethoxazole and oral nystatin, and empiric intravenous antimicrobial therapy during febrile granulocytopenic episodes were standard treatment for all patients . The use of trimethoprim/sulfamethoxazole did not appear to adversely affect donor marrow engraftment . Serious gram-negative bacillary and systemic Candida infections were uncommon . Although gram-positive bacterial infections were frequent, they were rarely associated with mortality . Aspergillosis emerged as the single most important infection, contributing to the death of nine patients . Cytomegalovirus diseases developed in 13 patients, seven of whom died . Patient age and chronic myelogenous leukemia were risk factors for the development of fatal infections . This study demonstrates that although certain serious infections can be controlled, there is a critical need for effective measures to prevent and treat aspergillosis and cytomegalovirus disease in these seriously compromised hosts. Infect Immun, 1983 Mar, 39(3), 1080 - 6 Macrophage activation in murine African trypanosomiasis; Grosskinsky CM et al.; African trypanosomiasis is accompanied by a profound general immunosuppression in which both suppressive T cells and macrophages (M phi) have been implicated . The present studies define changes in the M phi surface, endocytic and secretory properties, during the infection of mice by Trypanosoma brucei . Peritoneal M phi obtained after the control of the first wave of parasitemia displayed characteristics similar to those activated by intracellular pathogens, such as Mycobacterium bovis bacillus Calmette-Guerin, e.g., the enhanced expression of Ia antigen, decreased M phi-specific antigens, receptors mediating the pinocytosis of mannose-terminal glycoproteins, and an increased ability to secrete plasminogen activator, superoxide anion, and H2O2 . Some markers of macrophage activation persisted during the subpatent period before the recurrence of parasitemia, whereas others reverted to normal . Mature T cell function appears not to be essential for M phi activation by T . brucei since the infection of athymic nude mice also induced Ia antigens and plasminogen activator . These studies show that M phi activated by different pathways express common features which may contribute to immune dysfunction observed in trypanosomiasis, as well as in other infections. Biochem Biophys Res Commun, 1983 Feb 28, 111(1), 312 - 8 Structural and biosynthetic studies on linkage region between poly(galactosylglycerol phosphate) and peptidoglycan in Bacillus coagulans; Kaya S et al.; The HF treatment of teichoic acid-glycopeptide complexes isolated from lysozyme digests of Bacillus coagulans AHU 1366 cell walls gave a disaccharide, glucosyl beta (1 leads to 4)N-acetylglucosamine, along with dephosphorylated repeating units of the teichoic acid chain, galactosyl alpha (1 leads to 2) glycerol . Mild alkali treatment of the complexes yielded the disaccharide linked to glycopeptide, whereas direct heating of the cell walls at pH 2.5 yielded the same disaccharide linked to teichoic acid . The Smith degradation of the complexes revealed that the galactose residue is a component of backbone chain . Thus it is concluded that this disaccharide is involved in the linkage region between poly(galactosylglycerol phosphate) and peptidoglycan in cell walls . Membrane-catalyzed synthesis of this disaccharide on a lipid followed by transfer of glycerol phosphate from CDP-glycerol to the disaccharide-linked lipid in the absence or in the presence of UDP-galactose also supports this conclusion. J Biol Chem, 1983 Feb 25, 258(4), 2109 - 11 Respiration-dependent proton translocation in alkalophilic Bacillus firmus RAB and its non-alkalophilic mutant derivative; Lewis RJ et al.; Obligately alkalophilic Bacillus firmus RAB had a higher molar growth yield on L-malate (Ymal = 38 mg, dry weight/mmol of L-malate) than its non-alkalophilic mutant derivative, strain RABN (Ymal = 12 mg, dry weight/mmol of L-malate) . Measurements of respiration dependent proton translocation by the two strains in the presence of K+ and valinomycin showed that the alkalophile also has much higher H+/O stoichiometries (at pH 9.0) than does the mutant (at pH 7.0) . H+/O ratios for B . firmus RAB at pH 9.0 were as high as 13, with a frequently observed value of 9 . These high values were observed in the first phase of a set of biphasic curves for both oxygen consumption and proton ejection . At pH 7.0, both the wild type and the mutant exhibited H+/O ratios near 4 in a single phase of oxygen consumption and proton ejection . The results are consistent with suggestions that the alkalophilic respiratory chain is especially well adapted for effective energy transduction at alkaline but not neutral pH. Biochem J, 1983 Feb 15, 210(2), 411 - 7 Precursor processing during the maturation of a spore-coat protein in Bacillus megaterium KM; Stewart GS et al.; A protein of apparent mol.wt . 35000 that is extractable from the purified coat fraction of Bacillus megaterium KM spores is synthesized during sporulation as a precursor protein from which a 12-13 amino acid peptide is removed . Cleavage of this small peptide is delayed until 60-90 min after precursor synthesis and is concomitant with the morphological appearance of stage VI . The addition of chloramphenicol, subsequent to precursor synthesis, prevents the appearance of this late processing event . Two-dimensional non-equilibrium pH-gradient gel electrophoresis of the integument extract of forespores isolated at stage V from sporangia pulse-labelled with L-{35S}methionine 1 h before isolation, revealed both unprocessed and processed components . Similar analysis of total protein from the corresponding mother cells revealed only the unprocessed component in relatively small amounts, suggesting that, although the protein may be synthesized in the mother-cell compartment, processing may be restricted to the forespore . Peptide analysis by limited proteolysis was used to examine the relationship between the 35000- and a 17500-mol.wt . coat protein . The possible implications of limited proteolytic processing to maturation of the spore coat are discussed. Biochim Biophys Acta, 1983 Feb 15, 742(3), 617 - 22 Catalytic properties of Sepharose-bound L-alanine dehydrogenase from Bacillus cereus; Muresan L et al.; (1) L-Alanine dehydrogenase from Bacillus cereus was purified by a two-step chromatographic procedure involving Cibacron-Blue 3G-A Sepharose 4B-CL, and Sepharose 6B-CL, and immobilized on CNBr-activated Sepharose 4B . (2) Following immobilization via two of the six subunits, L-alanine dehydrogenase retained 66% of the specific activity of the soluble enzyme . The affinity of the immobilized enzyme for NH4+, pyruvate and L-alanine, was not different to that of the soluble form . The Km of the Sepharose-bound L-alanine dehydrogenase for pyridine coenzymes was 6-8-times higher than in the soluble case . (3) The stability of L-alanine dehydrogenase towards urea or thermal denaturation was increased by immobilization . (4) The incubation at 37 degrees C for 24 h of the immobilized L-alanine dehydrogenase with 3 M NH4Cl/NH4OH buffer (pH 9) released 70% of the enzyme . The specific activity and the affinity of the 'solubilized' L-alanine dehydrogenase for the pyridine coenzymes was the same as that obtained with the original, soluble L-alanine dehydrogenase. Anal Biochem, 1983 Feb 15, 129(1), 245 - 8 A method for the isolation of intact, viable forespores from Bacillus species using high pressure; Lindsay JA et al.; Large-scale preparation of Bacillus forespores was performed using nitrogen gas under high pressure to force an osmotically stable spheroplast suspension through a micrometer needle valve . Ninety-nine percent recovery of intact viable forespores at various stages of sporulation was achieved with a variety of Bacillus species . The isolated forespores are stable for up to 7 days when kept under nongerminating conditions. J Biol Chem, 1983 Feb 10, 258(3), 1960 - 7 Transcriptional and translational start sites for the Bacillus thuringiensis crystal protein gene; Wong HC et al.; The nucleotide sequence of the promoter region and part of the coding region of the crystal protein gene from Bacillus thuringiensis var . kurstaki HD-1-Dipel has been determined by analysis of a recombinant plasmid from Escherichia coli . The start points for transcription of the gene in B . thuringiensis and in the E . coli strain carrying the recombinant plasmid were located by S1 nuclease mapping . Two adjacent start sites were identified using RNAs synthesized during sporulation of B . thuringiensis: transcription was initiated from one site early in sporulation and from the other site in the middle of sporulation . A good correlation was found between the appearance of the crystal protein gene-specific RNA and the production of the protein, indicating that the gene is primarily under transcriptional control during sporulation . Parallel studies with the recombinant strain of E . coli revealed the presence of only a single species of gene-specific RNA, regardless of the growth phase of the cells; the crystal protein was produced at all stages of growth . The sequence for eight amino acids at the NH2 terminus of the crystal protein was determined and the corresponding coding sequence was located in the DNA sequence . A potential ribosome binding site of 11 nucleotides was found, located three nucleotides upstream from the initiator ATG codon . The deduced sequence for the first 333 amino acids of the crystal protein is presented. J Appl Bacteriol, 1983 Feb, 54(1), 131 - 4 A note on the microbial spoilage of undercooked chub-packed luncheon meat; Bell RG et al.; Contrary to expectations slight undercooking 968.5 degrees C instead of 70 degrees C for 90 min) dramatically increased the shelf-life of chub-packed luncheon meat stored at 25 degrees C . The pH of undercooked chubs fell rapidly to below 5.0 as a result of the growth of enterococci . The accumulated acid prevented the growth of Bacillus spores and gave the luncheon meat a not unpleasant tangy flavour . Degradative changes associated with the spoilage of commercially cooked chub-packed luncheon meat did not occur, even after 42 d storage . Apparently, post-cooking fermentation by enterococci can effectively convert a perishable product into a 'shelf stable' one by lowering the pH below 5.0. J Exp Med, 1983 Feb 1, 157(2), 807 - 12 Macrophage activation selectively enhances expression of Fc receptors for IgG2a; Ezekowitz RA et al.; After infection with bacillus Calmette-Guerin, peritoneal macrophages (Mo) display enhanced expression of FcR for both monomeric and complexed IgG2a, but not IgG2b . Isotype specificity of FcR can be reversed on nonactivated Mo by immune lymphokines, and IgG2a immune complexes are more effective triggers of the respiratory burst in activated Mo . Selective enhancement of IgG2a FcR by Mo activation could account for efficacy of homologous ab in mediating cytotoxicity in some systems. Dis Colon Rectum, 1983 Feb, 26(2), 129 - 32 Rectal malacoplakia in a patient with Hodgkin's disease . Report of a case and review of the literature; Yang CC et al.; A 37-year-old man was found to have classic malacoplakia of the rectum after three courses of chemotherapy for stage IV Hodgkin's disease . Sigmoidoscopy with excisional biopsy was performed because of rectal bleeding . The biopsy specimen of the rectal lesion showed focal extensive granulomatous changes with large macrophages containing round, dense Michaelis-Gutmann bodies . Electromicroscopy demonstrated calcifying spheres with laminated concertric structures (Michaelis-Gutmann bodies) and coliform bacillus in the cytoplasm of macrophages (Von Hansemann bodies) . Review of the literature revealed that association of malacoplakia with Hodgkin's disease has never been documented, although it has been reported to be associated with conditions such as malignancy, organ transplantation, collagen disease, and leukemia . The possible role of immune disturbance as an underlying cause of malacoplakia is discussed. Can J Microbiol, 1983 Feb, 29(2), 271 - 5 Alkaline extraction of toxin from spores of the mosquito pathogen, Bacillus sphaericus strain 1593; Davidson EW; Toxin was extracted from spores of the mosquito pathogen Bacillus sphaericus strain 1593 using 0.05 M NaOH . The molecular weight of this toxin was 35000-54000 . Toxic activity of this extract was resistant to a variety of enzymes including subtilisin, but was degraded by pronase . Antiserum produced to 1593 spore toxin neutralized spore toxin and cytoplasmic toxin activity, but did not react with Bacillus thuringiensis var . israelensis crystal toxin, nor did var . israelensis toxin antiserum react with B . sphaericus toxin . Crystal like parasporal inclusions accompanying the B . sphaericus 1593 spores were removed by NaOH extraction. J Biochem (Tokyo), 1983 Feb, 93(2), 469 - 77 Proline iminopeptidase from Bacillus megaterium: purification and characterization; Yoshimoto T et al.; Proline iminopeptidase {EC 3.4.11.5} was purified about 1,700-fold from cell free extract of Bacillus megaterium by a series of column chromatographies on DEAE-Toyopearl, PCMB-T-Sepharose and hydroxyapatite, and gel filtration on Toyopearl FW-55 . The purified enzyme still contained a minor contaminant as judged by disc gel electrophoresis . The enzyme was most active at pH 7.0 with Pro-beta-naphthylamide (Pro-2-NNap) as the substrate, and hydrolyzed Pro-X (X = amino acid, peptide, amide, and arylamide) bonds when the proline residue was at the amino terminal . The enzyme was completely inactivated by p-chloromercuribenzoate (PCMB), but was not inhibited by metal chelators, diisopropylphosphorofluoridate (DFP) and phenylmethanesulfonyl fluoride (PMSF) . The enzyme inactivated with PCMB was reactivated by adding 2-mercaptoethanol . From this result and the chromatographic profile on PCMB-T-Sepharose, the enzyme seems to be a sulfhydryl enzyme . The isoelectric point of the enzyme was 4.0 . The molecular weight of the enzyme was estimated to be 58,000 by gel filtration on Toyopearl and 60,000 by sodium dodecyl sulfate (SDS) gel electrophoresis, suggesting that the enzyme is a monomer. Proc Soc Exp Biol Med, 1983 Feb, 172(2), 260 - 4 Failure of athymic-nude mice sensitized with Bacillus Calmette-Guérin to produce interferon in response to purified protein derivative; Huygen K et al.; Unlike their +/+ and nu/+ littermates, homozygous athymic-nude (nu/nu) mice that had been sensitized with Bacillus Calmette-Guerin (BCG) failed to produce interferon (IFN) in response to purified protein derivative (PPD) . Likewise, spleen cells derived from BCG-sensitized nu/nu mice failed to produce IFN when exposed to PPD in vitro . These findings suggest that the T-cell subset that responds to specific antigens (i.e., PPD) is absent in athymic-nude mice. J Bacteriol, 1983 Feb, 153(2), 700 - 8 Effect of inhibitors of trypsin-like proteolytic enzymes Bacillus cereus T spore germination; Boschwitz H et al.; The germination of Bacillus cereus T spore suspensions is partially prevented by several inhibitors of trypsin-like enzymes . Leupeptin, antipain, and tosyl-lysine-chloromethyl ketone are effective inhibitors, whereas chymostatin, elastatinal, and pepstatin are inactive . A synthetic substrate of trypsin, tosyl-arginine-methyl ester, also inhibits germination . Its inhibitory effect decreases as a function of incubation time in the presence of spores and is abolished by previous hydrolysis with trypsin . Germinating, but not dormant, spore suspensions hydrolyze tosyl-arginine-methyl ester; its hydrolysis is insensitive to chloramphenicol, sulfhydryl reagents, and EDTA . A crude extract of germinated B . cereus spores contains a trypsin-like enzyme whose activity, as measured by hydrolysis of benzoyl-arginine p-nitroanilide, is sensitive to germination-inhibitory compounds such as leupeptin, tosyl-arginine-methyl ester, and tosyl-lysine-chloromethyl ketone . Spore suspensions exposed to the above inhibitors under germination conditions lose only part of their heat resistance and some 10 to 30% of their dipicolinic acid content . Part of the germinating spore population becomes "phase grey" under phase optics . Based on a study of the inhibition of germination by protease inhibitors and the activity of a protease in germination spores and spore extracts, it is suggested that the activity of a trypsin-like enzyme may be involved in the mechanism of the breaking of dormancy in spores of B . cereus T. Cancer Res, 1983 Feb, 43(2), 585 - 91 Retinoid feeding, hormone inhibition, and/or immune stimulation and the genesis of carcinogen-induced rat mammary carcinomas; Welsch CW et al.; Female Sprague-Dawley rats were treated at 53 days of age with a single intubation of 7,12-dimethylbenzanthracene (DMBA) . Three days after carcinogen treatment, the animals were treated with retinyl acetate (RA) (at 3 dietary levels), hormone inhibition (HI) {tamoxifen (1-rho-beta-dimethylaminoethoxyphenyl-trans-1,2-diphenylbut-1-ene) plus 2 bromo-alpha-ergocryptine}, and/or immune stimulation (methanol-extracted residue of Bacillus Calmette-Guerin, cell wall skeleton of Nocardia rubra, or cell particulate of DMBA-induced rat mammary carcinomas plus Freund's complete adjuvant) . RA at 0.6 or 1.0 mM concentrations per kg diet significantly reduced the incidence of mammary carcinomas; 0.2 mM concentrations of RA per kg diet did not affect tumor incidence . HI also significantly decreased mammary carcinoma incidence, an effect which was significantly enhanced by all 3 dietary levels of RA . Immune stimulation by methanol-extracted residue of Bacillus Calmette-Guerin or cell wall skeleton of Nocardia rubra did not affect mammary carcinoma incidence when administered either alone or in combination with RA and/or HI . The cell particulate of DMBA-induced rat mammary carcinomas plus Freund's complete adjuvant significantly reduced mammary carcinoma incidence in rats fed RA but did not affect mammary carcinoma incidence in placebo-fed rats or in rats treated only with HI . However, in rats treated with the triple combination of cell particulate of DMBA-induced rat mammary carcinomas plus Freund's complete adjuvant, RA, and HI, no mammary carcinomas were observed for the duration of treatment (20 weeks after DMBA administration) . Although HI was always superior to RA feeding in the prophylaxis of this neoplastic process, a significant synergism between these two treatments was consistently observed . This distinct synergism was observed even when using the low dietary level of RA, an amount of RA which by itself was ineffective in the suppression of mammary carcinogenesis . With but one exception, immune stimulation did not significantly influence this carcinogenic process, either when administered alone or when administered to rats with a reduced mammary carcinoma burden, i.e., animals treated with RA and/or HI. Appl Environ Microbiol, 1983 Feb, 45(2), 451 - 5 Automatic diluter for bacteriological samples; Trinel PA et al.; The described apparatus, carrying 190 tubes, allows automatic and aseptic dilution of liquid or suspended-solid samples . Serial 10-fold dilutions are programmable from 10(-1) to 10(-9) and are carried out in glass tubes with screw caps and split silicone septa . Dilution assays performed with strains of Escherichia coli and Bacillus stearothermophilus permitted efficient conditions for sterilization of the needle to be defined and showed that the automatic dilutions were as accurate and as reproducible as the most rigorous conventional dilutions. J Bacteriol, 1983 Feb, 153(2), 1083 - 7 Q beta replicase containing a Bacillus stearothermophilus elongation factor; Stringfellow L et al.; We purified Q beta replicase containing EF-Ts from Bacillus stearothermophilus in place of the homologous polypeptide from Escherichia coli . The hybrid enzyme was fully active in the transcription of a variety of templates . It was found to be qualitatively similar to native Q beta replicase with respect to a variety of parameters which measure the efficiency of initiation of RNA synthesis . The results demonstrated that Q beta replicase can tolerate substantial alterations in the EF-Tu X Ts component of the enzyme . These alterations resulted in only minor perturbations of catalytic properties. Am J Pathol, 1983 Feb, 110(2), 182 - 92 Macrophage polykaryon formation in vitro by peritoneal cells from mice given injections of sodium periodate; Weinberg JB; Peritoneal macrophages from mice that have received two separate intraperitoneal injections of the sterile, soluble oxidant NaIO4 form macrophage polykaryons (MPs) in vitro, but peritoneal macrophage from untreated, peptone-treated, or mice infected with bacille Calmette-Guerin (BCG) do not . The polykaryons are noted after 18-24 hours of culture and continue to form over a 60-72-hour period . The MPs do not form if the macrophage density is less than 4 x 10(3)/sq mm . The polykaryons appear in vitro only in cultures with less than or equal to 1-5 ng/ml lipopolysaccharide (LPS) (amounts of LPS that commonly contaminate culture medium and serum) . Hydrocortisone hemisuccinate (2.6 x 10(-9) M) inhibits MP formation in vitro . Lymphocytes do not influence the polykaryon formation, and supernatants from MP cultures do not cause fusion of other macrophages . Microcinephotography demonstrates fusion of the macrophages to form the large polykaryons, which are less motile than uninuclear macrophages . The polykaryons assume different forms; generally, the nuclei (mean, 16.8 nuclei/MP; range, 2-137 nuclei/MP) are centrally located, and the nuclear chromatin of all nuclei appears similar . The MPs phagocytize polystyrene spheres and glutaraldehyde-treated erythrocytes to the same degree as do uninuclear macrophages when determined as particles per nucleus (phagocytic index), but their phagocytic index of IgG-coated erythrocytes is decreased . Peritoneal macrophages from mice given double injections of NaIO4 are nontumoricidal in the absence of LPS, but LPS, in amounts sufficient to inhibit polykaryon formation, renders the macrophages tumoricidal . Populations of macrophages containing MPs formed over 3 days of cultures also respond to LPS or macrophage activating factor (MAF) to demonstrate enhanced tumoricidal activity. J Gen Microbiol, 1983 Feb, 129 (Pt 2), 311 - 9 Growth defects of Escherichia coli cells which contain the gene of an alpha-amylase from Bacillus coagulans on a multicopy plasmid; Willemot K et al.; An alpha-amylase gene from Bacillus coagulans has previously been cloned in Escherichia coli and shown to direct the synthesis of an enzymically active protein of 60,000 Dal (Cornelis et al., 1982) . In one particular E . coli host, strain HB101, amylase was found to accumulate in the periplasmic space . To study the processing and the location of the amylase, plasmid pAMY2 was introduced into E . coli 188 which is a strain constitutive for alkaline phosphatase, a periplasmic marker, and for beta-galactosidase, a cytoplasmic marker . Abnormally large amounts of both alpha-amylase and beta-galactosidase were found in the culture fluid of cells grown in rich medium . Furthermore a severe growth defect was found when cells containing pAMY2 were grown in maltose and glycerol media, while the ability to grow on glucose remained normal . This defect could be reversed by two types of spontaneous mutations . Mutations in the first class are located on the plasmid and correspond to the insertional inactivation of the amylase gene by IS1 . Mutations in the second class are located on the host chromosome . These results suggest that the synthesis and export of B . coagulans alpha-amylase is deleterious to E . coli, especially in media containing maltose or glycerol as sole carbon source. Biochim Biophys Acta, 1983 Jan 25, 755(2), 253 - 6 Induction of phosphoribomutase in Bacillus cereus growing on nucleosides; Ipata PL et al.; In this paper we show that phosphoribomutase is induced in Bacillus cereus by the same metabolizable purine and pyrimidine ribonucleosides previously shown to induce the purine nucleoside phosphorylase (Tozzi, M.G., Sgarrella, F . and Ipata, P.L . (1981) Biochim . Biophys . Acta 678, 460-466) . The mutase allows ribose 1-phosphate formed from nucleosides to be utilized by the cell through the pentose cycle, upon transformation to ribose 5-phosphate . The equilibrium constant of the mutase reaction is towards ribose-5-phosphate formation . The coordinate induction of the two enzymes completes the picture of the molecular events leading to the utilization of the sugar moiety of purine nucleotides and nucleosides as an energy source (Mura . U., Sgarrella, F . and Ipata, P.L . (1978) J . Biol . Chem . 253, 7905-7909). Schweiz Med Wochenschr, 1983 Jan 22, 113(3), 71 - 4 {Problems of tuberculosis in the world today}; Chaulet P; A century after Robert Koch's discovery of the tubercle bacillus the world total of tuberculosis sufferers is estimated at 20 million, 10 million of whom are contagious . Contrary to a widespread opinion, tuberculosis has not regressed over the last 30 years because it chiefly affects the Third World, in which countries the population doubles every 20 or 25 years while economic resources remain static for the majority of the population . Tuberculosis will be a serious problem of world health in the year 2000 . These facts should stimulate physicians to exploit the technical possibilities to the full, with stress upon detection and chemotherapy of tuberculosis . A health program will need to be established for these nations which will make it possible to identify and treat the maximum number of sources of infection . All the investigations and efforts at international cooperation in the field of tuberculosis should be directed towards improving or intensifying national antituberculosis programs. Schweiz Med Wochenschr, 1983 Jan 22, 113(3), 75 - 9 The contagiosity of tuberculosis; Riley RL; Although a few advanced thinkers understood the contagious nature of tuberculosis in the middle of the last century, more precise concepts of transmission awaited Pasteurs's demonstration in 1862 that organisms exist in the air and Koch's identification of the tubercle bacillus in 1882 . At the end of the last century Cornet presented evidence that tuberculosis was dust-borne, and Flugge presented equally convincing evidence that tuberculosis was droplet-borne . Chapin, in 1910, summarized evidence against the prevaling belief in airborne transmission of many common infections but made an exception of tuberculosis . Chapin emphasized transmission by direct contact (touching) . In the 1930's William F . Wells introduced the droplet nucleus hypothesis . In the 1950's droplet nuclei were shown to be the carriers of infection from tuberculosis patients to guinea pigs breathing air vented from the patients' ward . Subsequent evidence has indicated that direct contact is unimportant, although close proximity facilitates airborne transmission . Attempts to control the spread of tuberculosis by air disinfection or by immunization have been disappointing . Specific antituberculosis drugs, widely used since the 1950's, cause patients to become noninfectious for others very rapidly, even before organisms in the sputum disappear . Chemotherapy is now recognized as the corner stone of contagion control. Nucleic Acids Res, 1983 Jan 11, 11(1), 203 - 10 Calorimetric measurements of the destabilisation of a ribosomal RNA hairpin by dimethylation of two adjacent adenosines; Heus HA et al.; Fragments of 16S ribosomal RNA from E . coli and B . stearothermophilus, respectively comprising the 49 and 52 3' terminal nucleotides have been studied thermodynamically using high sensitivity differential scanning calorimetry . The fragments were isolated after cleavage of 16S rRNA in the ribosome by the bacteriocin cloacin DF13 . Comparison of the thermal properties of the E . coli fragments with those derived from a kasugamycin resistant mutant, which specifically lacks dimethylation of two adjacent adenosines was employed to study the effect of the methylgroups on the thermal stability . Both E . coli species exhibit similar complex melting patterns with several transitions . Overall molar transition enthalpies are equal and do not depend significantly on buffer conditions (120 kcal/mol at 15 mM Na+ to 136 kcal/mol at 215 mM Na+) . However, the transition with the highest Tm, corresponding to unfolding of a nine basepair central helix is lowered by the dimethylation of the adenines in the four-membered loop . This decrease amounts to 4 degrees C at 15 mM Na+ and 2 degrees C at 215 mM Na+ . The corresponding nine basepair long hairpin in the Bacillus fragment melts at a temperature of 70 degrees C at 15 mM Na+ . This Tmax is much higher than expected on the basis of the sequence in the hairpin. J Biol Chem, 1983 Jan 10, 258(1), 35 - 7 Oxidative phosphorylation by isolated membrane vesicles from Bacillus megaterium and its uncoupler-resistant mutant derivative; Guffanti AA et al.; ATP synthesis was studied in ADP + Pi-loaded, right-side-out membrane vesicles from Bacillus megaterium and its uncoupler-resistant mutant strain, C8 . Upon energization with ascorbate/phenazine methosulfate, more ATP synthesis was observed in C8 vesicles than in those from the wild type . ATP synthesis by C8 vesicles was more resistant to low levels (0.5-1.0 microM) of carbonyl cyanide m-chlorophenylhydrazone than was synthesis by wild type vesicles, whereas synthesis by both preparations was completely inhibited by N,N'-dicyclohexylcarbodiimide . Upon energization by a valinomycin-induced potassium diffusion potential, vesicles from the wild type strain synthesized more ATP than vesicles from C8, but that synthesis was still lower than observed with electron donors . The results indicate that the characteristic bioenergetic properties exhibited by whole cells of C8 are retained in a vesicle system and thus cannot be attributed to a cytoplasmic, substrate level activity . Interestingly, lipophilic cations that were efficacious in measuring the transmembrane electrical potential of whole cells appeared to accurately measure artificially generated potentials across vesicle membranes, but were not taken up upon addition of ascorbate/phenazine methosulfate. Rev Fr Mal Respir, 1983, 11(2), 79 - 110 {Problems posed by tuberculosis in the world in 1982}; Chaulet P et al.; One hundred years after the discovery of the tubercle bacillus the struggle against tuberculosis still poses numerous problems on a world scale . Each year, 10 million new cases appear: nearly five million are highly contagious; 95% of these cases come from developing countries . The official notification system currently in force is imperfect but a reckoning of the annual infection risk, from a methodical enquiry into tuberculous disease gives exact information on the size of the problem and its evolutionary time scale in each country . This is the best epidemiological index currently available . The technical problems of the struggle against tubercle are resolved in theory: the selective detection of groups at risk, centered on a network of bacteriology laboratories enables identification of all the sources of infection; chemotherapy, becoming safer and safer, with a duration reduced to 6 or 9 months at the maximum and curing practically 100% of the patients; properly performed BCG vaccination reduces the risk of infantile tuberculosis . Operational problems remain the most serious . The greatest difficulties are seen in countries with the highest prevalence: they are linked to a global shortage of resources but also at times a poor utilisation of the resources available . A concerted international action could render antituberculous drugs and the appropriate sanitary technology to all countries . With a judicious use of available information and means each country could today achieve an effective national antituberculous programme. Am Rev Respir Dis, 1983 Jan, 127(1), 39 - 41 Endobronchial pH . Relevance of aminoglycoside activity in gram-negative bacillary pneumonia; Bodem CR et al.; The in vitro bactericidal activity of certain antimicrobials, especially aminoglycosides, is markedly diminished at an acid pH . In an attempt to correlate this factor with the poor response of gram-negative bacillary pneumonia to aminoglycoside therapy, the endobronchial pH of selected groups of subjects was measured via a bronchoscopically directed pH electrode . The average peripheral, i.e., subsegmental, endobronchial pH of normal persons, patients with chronic lung disease, and patients with pneumonia was 6.58 +/- 0.06, 6.62 +/- 0.10, and 6.61 +/- 0.06, respectively . The average central airway, i.e., major bronchi or tracheal, pH at 6.64 +/- 0.07 did not vary significantly from that of peripheral airways . The presence of pneumonia in individual bronchi was associated with a significantly lower pH than that in noninfected bronchi: 6.48 +/- 0.12 versus 6.69 +/- 0.13 (p less than 0.05) . This relatively acid environment appears exaggerated within pneumonic airways and may contribute toward decreasing the effectiveness of aminoglycosides and certain other antimicrobials used in treating lung infection. Am Rev Respir Dis, 1983 Jan, 127(1), 35 - 8 Mycobacterium simiae . Clinical features and follow-up of twenty-four patients; Bell RC et al.; The charts of twenty-four patients from whom Mycobacterium simiae was isolated from the sputum were reviewed and the patients seen in follow-up examination when possible . They were divided into 3 groups: 2 patients were felt to have had definite infection with M . simiae, 3 were felt to have had probable infection, and 19 showed no evidence of infection during follow-up for as long as 6 yr . All patients in the study had underlying pulmonary abnormalities . The results of PPD skin tests were negative in patients without evidence of tuberculosis . The patients without evidence of M . simiae infection were found to have had negative initial Acid Fast Bacillus smears, fewer sputum cultures positive for M . simiae, and lighter yields from cultures of M . simiae compared with those in the patients with M . simiae infection . We conclude that M . simiae is a nontuberculous mycobacterium capable of causing progressive granulomatous lung infection, but that it may also be identified as a causal isolate from the sputum of susceptible persons . Antituberculosis chemotherapy should not be employed in this latter group. Zentralbl Mikrobiol, 1983, 138(1), 9 - 15 Growth and nitrogen fixation by cultures of Bacillus sp . S 77 g, Pseudomonas sp . S 44 b, and Klebsiella sp . S 145 m, isolated from Egyptian soils; Shawky BT; Study of the behaviour of representative aerobic freeliving dinitrogen-fixers, other than azotobacters, with respect to different carbon sources, showed that a broad range of compounds is accessible to them . This group of nitrogen-fixers can assimilate molecular nitrogen as a result of utilization of carbohydrates, alcohols, and organic acid salts . The highest nitrogen-fixation productivity, reaching 20.6-27.3 mg of nitrogen per g of carbon utilized, was observed on medium containing glucose as the carbon and energy source . Supplementary growth factors added to the N-deficient medium in the form of small amounts of yeast extract stimulated their growth and fixation of atmospheric nitrogen . This stimulation is particularly evident in mixed cultures . The nature of the mutual influences of micro-organisms in these cultures remains fairly obscure and further inquiry is necessary to elucidate this problem . A medium containing 1% glucose and Difco yeast extract to produce a bound nitrogen concentration of 10 ppm appeared to be the most favourable for the cultivation and N2-fixation of this group of microorganisms. Plasmid, 1983 Jan, 9(1), 98 - 103 Evidence for plasmid-associated crystal toxin production in Bacillus thuringiensis subsp . israelensis; Faust RM et al.; Three crystalliferous (Cry+) strains of Bacillus thuringiensis subsp . israelensis (serotype 14) that produce parasporal protein crystals toxic to dipteran larvae and several acrystalliferous (Cry-) mutants, either induced or spontaneously derived from a single Cry+ parent, were examined for the presence of covalently closed circular (CCC) DNA in attempts to correlate toxin production with the presence of a specific plasmid . The plasmid profiles of both Cry+ and Cry- variants were analyzed by both a cleared lysate- and a modified Eckhardt lysate-electrophoresis technique . All of the Cry- mutants derived from the Cry+ parental strain had lost a 4.0- to 4.4-megadalton (Mdal) plasmid . Bioassay data confirmed loss of toxin production by the Cry- variants . All three Cry+ strains, including the parent of the Cry- strains, contained CCC plasmids DNAs of the following approximate molecular weights: 4.0 to 4.4, 5.2 to 6.0, and 11.4 to 13.0 Mdal . One Cry+ strain contained an additional CCC plasmid of 6.7 to 7.2 Mdal . The plasmid patterns for several Cry- derivatives differed in other respects from the pattern for their parent strain . The various Cry+ and Cry- strains could be distinguished either by phenotypical differences in antibiotic sensitivity, crystal production, and toxicity, or by differences in their plasmid profiles. Folia Microbiol (Praha), 1983, 28(1), 41 - 5 Antimicrobial activity of 3-methyl-5-furfurylidene-2-thioxo-4-thiazolidones; Awasthi LP et al.; Substituted 3-methyl-5-furfurylidene-2-thioxo-4-thiazolidones 1-17 were tested for their action against Bacillus pumilus, B . brevis and B . megaterium and against Aspergillus flavus, A . fumigatus and A . niger . Most of the compounds possessed an activity against both bacteria and fungi . These compounds were also tested for their effectiveness as virus inhibitors employing Nicotiana glutinosa as assay host against tobacco mosaic virus . Most of them exhibited potential antiviral activity both in vitro and in vivo. J Urol, 1983 Jan, 129(1), 29 - 32 Chemoimmune prophylaxis of superficial bladder cancer; Adolphs HD et al.; Chemoimmune prophylaxis with intravenous cyclophosphamide and intravesical as well as systemic bacillus Calmette-Guerin treatment was begun after complete tumor resection in 90 patients with superficial urothelial bladder cancer . Compared to a historical patient control group treated by tumor resection alone a distinct decrease in the recurrence rate was noted in the immune treated group, which was most marked during the first 12 months postoperatively . The side effects of this treatment were tolerable . Our results are discussed with regard to the reported findings in the literature . Possible immune biological mechanisms of the tumor protection achieved by cyclophosphamide and bacillus Calmette-Guerin are suggested. J Assoc Off Anal Chem, 1983 Jan, 66(1), 176 - 9 Determination of penicillin G, ampicillin, and cephapirin residues in tissues; Vilim AB et al.; A cylinder plate microbiological method was developed for the rapid, quantitative determination of penicillin G, ampicillin, and cephapirin in animal tissues . The method uses agar plates seeded with stable spores of Bacillus stearothermophilus var . calidolactis and incubated 4 h at 64 degrees C . Standard curves were obtained for the following ranges of concentration of antibiotics in tissues: 0.02-0.32 IU penicillin G/g, 0.0125-0.2 micrograms ampicillin/g, and 0.02-0.32 micrograms cephapirin/g . The proposed method is suitable not only for penicillin residue analysis, for which the sensitivity has been greatly improved compared with the Sarcina lutea method, but also for depletion studies on these antibiotics, which are commonly used to treat diseases in food-producing animals. Nahrung, 1983, 27(3), 257 - 63 {Microbiological degradation of glucosinolates in defatted rapeseed meal}; Huber J et al.; During the degradation of thioglucosides in defatted rape seed meal (RES) microorganisms were found, whose ability to degrade glucosinolates (GSL) and vinyl thio-oxazolidone (VTO) was not known so far . The isolated microorganisms are two strains of bacteria of the species Bacillus cereus and the yeast Trichosporon cutaneum . The degradation of GSL and VTO in the cultural broths by the Bacillus cereus strains was the more complete the more other Gram-negative bacteria from RES were additionally present . Drinking tests with Wistar rats showed that here is a relation between the watersoluble and mainly bitter toxin substances of rape and their influence on the increase of the animals body weight. Bibl Haematol, 1983, (49), 79 - 93 Incidence and clinicopathological significance of DIC in autopsy cases; Tanaka K et al.; Disseminated intravascular coagulation (DIC) was examined pathologically in 4906 consecutive autopsy cases during the last 11 years . The cases having pathologically confirmed DIC showing microthrombi in three or more organs were 88 . Of the underlying diseases for these cases, malignant tumor was found in 40 cases and diseases of hematopoietic organs in 19 . Of the cases with malignant tumor, 11 had gastric cancer, 7 had lung cancer, and 4 had pancreatic cancer . Thirty-three of the 40 cases with malignant tumor showed metastasis in two or more organs . Cases with pathologically confirmed or suspected DIC that had microthrombi in one or more organs were 319 . As for the incidence of pathologically suggestive DIC in each disease, the incidence of malignant tumor was 7.3% and that for diseases of the hematopoietic organ was 10.6% . Infection is an important underlying condition, especially gram-negative bacillus septicemia which may play an important role in the development of DIC . An increase in the number of white blood cells appears to be one of the causative conditions of DIC . Kidney is involved most frequently by the deposition of microthrombi, and 27 out of 88 cases show ischemic lesions induced by intravascular coagulation . There were 109 cases having clinically diagnosed or suspected DIC, but 67 cases showed no microthrombus formation . On the other hand, 63 among 4,797 cases with clinically unsuspected DIC revealed microthrombus formation in three or more organs by the postmortem examination. Zentralbl Mikrobiol, 1983, 138(7), 553 - 63 Utilization of fodder yeast and agro-industrial by-products in production of spores and biologically - active endotoxins from Bacillus thuringiensis; Salama HS et al.; A number of newly-devised fermentation media were evaluated with respect to their ability to support sporulation and biosynthesis of endotoxins by strains of Bacillus thuringiensis that are biologically active against Spodoptera littoralis, Heliothis armigera, and Spodoptera exigua . Fodder yeast from dried cells of Saccharomyces cerevisiae could be used as a complete mono-component medium for production of highly active spore-delta-endotoxin complexes from B . thur., vars . entomocidus, kurstaki and galleriae . Highest sporulation titers were obtained at 2% fodder yeast concentration with endotoxin yields ranging between 7 to 9 grams per liter of medium . Ground horse beans and kidney bean seeds could also be used successfully as complete media for sporulation and endotoxin production . Extracts of potato tubers and sweet potato roots were efficient media for active endotoxin production from B . thur . var . kurstaki, although the obtained yields were much lower than those produced in fodder yeast media . The utilization of fish meal, cotton seed meal, and residues of chicken from the slaughter-house as media for the production of endotoxins active against Spodoptera littoralis, was not successful . On the other hand, minced citrus peels, ground seeds of dates, and wheat bran could be successfully used in combination with fodder yeast as media for production of endotoxins, active against Heliothis armigera and Spodoptera exigua . Re-utilization of culture supernatants in a second fermentation cycle after supplementation with some nutrients gave promising results with some of the strains tested . The data obtained are discussed in view of their feasibility of application. Nauchnye Doki Vyss Shkoly Biol Nauki, 1983, (9), 21 - 6 {Mechanism of flagellin polymerization in Bacillus brevis}; Novikov VV et al.; The optical properties of acid-dissociated and tetranitromethane-modified flagellin has been studied by circular dichroism (CD) techniques . The flagellins have the same CD spectra which do not change over the 2.9-10.0 range . The spectra character depends on pH in the presence of polyethylene glycol (PEG) and ammonium sulphate which accelerate polymerization . As the content of PEG goes up to 20% and ammonium sulphate to 1 M, the value of the molecular ellipticity at 222 nm ({O}222) of the both flagellins considerably increases at pH 4.3-10.0, however {O}222 does not achieve the value for bacterial flagella . PEG and ammonium sulphate addition at pH 2.9-3.9 gives less dramatic increase of the {O}222 value . It has been concluded that the changes in the CD spectra at pH 4.3-10.0 is a result of conformational rearrangements in flagellin before it incorporates into the flagella. Z Erkr Atmungsorgane, 1983, 161(1), 107 - 16 {The strategy of tuberculosis control in East Germany after 1982}; Herrmann H; 100 years after ROBERT KOCH'S detection of the tubercle bacillus the tuberculosis situation in GDR has so improved due to the progress in tuberculosis control, systematic activities in prevention, casefinding, and care and due to the comprehensive social measures that tuberculosis cannot be any more regarded as a "Volkskrankheit" . The reduction of the annual infection rate to less than 0.1%, the decrease of infectious tuberculosis to less than 5/100,000 inhabitants, the shortening of the periods of chemotherapy and the further regression of relapses has made it possible to rationalize tuberculosis control by giving up indiscriminate mass x-ray examinations of the whole population in favour of the supervision of endangered person groups, by reducing the extent of risk groups according to stricter criteria, by making shorter the follow-up of the several forms of tuberculosis . Primary health care services have to be involved still more into tuberculosis control, especially in casefinding and outpatient treatment. Experientia Suppl, 1983, 46, 243 - 7 Advances in microbial protoplasts (Bacillus licheniformis beta-lactamase and the protoplast surface); Lampen JO; The changing nature of the research reported at these Symposia over the past two decades is noted, especially the current emphasis on plant protoplasts . Improvements in procedures for the preparation and regeneration of microbial protoplasts are summarized and the definition of optimal conditions for fusion and genetic transfer . The efficiency of fusion now attainable has led to considerable advances in the genetics of yeasts and mycelial fungi and of bacteria . In addition, the synthesis and secretion of beta-lactamase (penicillinase) in Bacillus licheniformis is described in order to illustrate how protoplasts can be utilized to investigate 1) the form in which the new enzyme emerges from the membrane surface, 2) the importance of accessibility in processing of the emerging chain and 3) the role of cell wall structure in the induction of beta-lactamase synthesis. Zentralbl Mikrobiol, 1983, 138(6), 475 - 85 Correlation between alpha-amylase production and sporulation in Bacillus licheniformis CUMC 305 with respect to the effect of some carbohydrates and phenylmethylsulfonyl fluoride treatment; Krishnan T et al.; The existence of any relationship between extracellular alpha-amylase production and sporulation in Bacillus licheniformis CUMC 305, when grown in the presence of different carbohydrates, was investigated . It was noted that alpha-amylase production in the organism was almost complete during the period of maximum sporulation, irrespective of the carbon source used . The effect of a sporulation inhibitor, phenylmethylsulfonyl fluoride at a concentration of 8 micrograms/ml, on enzyme secretion was an interesting feature . It was observed that alpha-amylase production as well as sporulation deteriorated, if the inhibitor was added to the external growth medium during the period extending from mid-logarithmic phase to mid-stationary phase of the growth cycle; contrary to it, there was no change in the pattern of alpha-amylase production or sporulation when added either earlier or later than this period . It may be concluded that the extracellular pool was the principal yield of alpha-amylase in the organism, since washing experiments and subsequent treatment of the intact cells with Tris hydroxymethyl aminomethane hydrochloride buffer or lysozyme, surfactant (Tween-80), or lysozyme-Tween mixture failed to extricate any substantial quantities of the enzyme . The intracellular pool of enzyme, obtained by disrupting the cells, was insignificant compared to the extracellular pool. Lepr India, 1983 Jan, 55(1), 91 - 4 Protease inhibitors activity in lepromatous leprosy and lepra reaction; Yemul VL et al.; Serum alpha one antitrypsin levels were measured in 50 healthy age and sex matched controls with 45 lepromatous leprosy cases and 5 cases of lepra reaction . It was noted that the mean level in healthy controls was 281.00 mg%, while the mean levels in LL patients was 421.00 mg% and in LR 570.00 mg% . The elevation of Alpha one antitrypsin was statistically significant in LL patients . It is possible that the rise is a reaction to release of proteases and or higher complement activity, which are the results of a high bacillary loading to formation of immune complexes. Lepr India, 1983 Jan, 55(1), 29 - 38 Lymphocyte subpopulations in mice infected with Mycobacterium leprae; Ganguly NK et al.; T and B cells were quantitated from the spleen of M . leprae infected mice and correlated with bacillary count in the foot pad . Lymphocyte transformation with PHA and M . leprae (armadillo) antigens was also studied during different months of infection . T cell counts dropped gradually but significantly throughout the course of infection . B cells had a concomitant rise up to 6 months and then registered a fall as compared to the initial control figures . Transformation of lymphocytes with PHA fell significantly after 4th month til the end of the experiment, whereas stimulation index for armadillo antigen rose gradually from 3rd month onwards to reach a peak at 6th month and then fell till the end of the experiment with increasing bacterial population whereas B-cell counts showed little change. Experientia Suppl, 1983, 46, 149 - 54 Chromosome interactions and expression in fused Bacillus protoplasts; Hotchkiss RD et al.; Heterodiploid bacteria can be regenerated from fused mixed protoplasts . In both B . megaterium and B . subtilis, early selective effects during regeneration may strongly affect the phenotype of the products . Among the products are diploid prototrophs, whose stability is still in doubt . It is not known whether the prototrophy results from complementation or from recombination during the selection . In the case of B . subtilis, regeneration in a permissive, uncrowded and non-selective environment leads to production of rather large yields of heterodiploids, either biparental or recombinant . These seem to be only partially stable, but while remaining diploid they give rise to a wide variety of genetic recombinants . In general rather few prototrophs are found when selection is applied to the bacterial diploids . The phenotypic properties of B . subtilis diploids reveal incomplete expression of the demonstrable gene inventory . Biparental diploids appear commonly to show the phenotype of only one of the contained parent types, as if one chromosome remains unexpressed . For diploid recombinants data are insufficient to reveal the status of the chromosomes . Non-expression is believed to be due to structural complexity of particular chromosomes, or their parts . Recombination occurring within bacterial heterodiploid clones appears to satisfy many of the expectations for genetic recombination in eukaryotes--including that of being in part at least classically reciprocal, with intraclonal reciprocals in equivalent numbers . While short map intervals show reduced recombination, large numbers of recombinations (one to two thirds) involve exchanges at or near the terminus and origin of bidirectional replication.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Gen Genet, 1983, 189(3), 475 - 8 Kasugamycin resistant mutants of Bacillus stearothermophilus lacking the enzyme for the methylation of two adjacent adenosines in 16S ribosomal RNA; Van Buul CP et al.; Several mutants of B . stearothermophilus have been isolated that are resistant to the antibiotic kasugamycin . One of these is shown to lack dimethylation of two adjacent adenosines in the 16S ribosomal RNA . All mutants that were analyzed biochemically lack the enzyme that is able to methylate this site . Ribosomal sensitivity and resistance to kasugamycin in B . stearothermophilus is therefore, like in E . coli, closely connected with dimethylation of the adenosines. Med Pediatr Oncol, 1983, 11(2), 79 - 90 Intermittent combination chemotherapy with or without bacillus Calmette-Guérin for treatment of acute lymphoblastic leukemia of childhood; Odom LF et al.; Seventy-four children ranging in age from 6 months to 17.5 years with acute lymphoblastic leukemia newly diagnosed between 1976 and 1979 were entered on a study incorporating intermittent chemotherapy with or without the addition of bacillus Calmette-Guerin (BCG) . The chemotherapy program consisted of induction with vincristine, dexamethasone, and intrathecal methotrexate, intensification with adriamycin and asparaginase, central nervous system treatment with cranial irradiation and intrathecal methotrexate, and continuation treatment with 5-day courses of combination chemotherapy administered every three weeks . The first phase of continuation therapy incorporated vincristine, adriamycin, 6-mercaptopurine, and dexamethasone . In the second phase, oral methotrexate was substituted for the adriamycin in non-T-cell patients; in T-cell patients, cytosine arabinoside or cyclophosphamide and methotrexate in alternating cycles were substituted for the adriamycin and asparaginase was added . Total duration of therapy was approximately 2.5 years . Connaught BCG was administered by Heaf gun on days 8 and 15 of each 3-week cycle for the first 8 months of treatment in approximately one-third of the patients . Actuarial disease-free survival with a median follow-up of 59 months shows no difference in outcome between the BCG and non-BCG poor-risk patients . However, there is an improvement in disease-free survival of BCG-treated good- and average-risk girls (P = 0.04) . While patients were actively receiving BCG there was also a trend toward the development of fewer significant infections than when patients were not receiving BCG (P = 0.85) . Toxicities from BCG administration included satellite rashes, local tenderness, lymphadenopathy, secondary infection, and residual scars . Overall disease-free survival by actuarial analysis is 60% at 6 years; for patients with unfavorable prognostic features it is 40% . In this trial the addition of BCG prolonged the disease-free survival of girls with good- and average-risk prognostic features and also may have decreased the susceptibility to infection while it was being administered . However, the benefit does not appear sufficient to warrant its routine use, especially in view of the toxicities encountered. Cancer Immunol Immunother, 1983, 15(1), 17 - 22 Natural cell-mediated cytotoxicity in normal human peripheral blood lymphocytes and its in vitro boosting with BCG; Mandeville R et al.; Natural cell-mediated cytotoxicity (NCMC) against K-562 human erythroleukemic cells was monitored in an overnight chromium release assay using normal human peripheral blood lymphocytes (PBL) as effective cells . Two hundred and ten normal individuals were tested from 3 to 24 times over a period of 3 years . The level of NCMC was shown to vary from 4% to 46% lysis at an effector-to-target cell ratio of 5/1; males had higher levels of activity than females (P less than 0.001) . A group of individuals with low natural killer (NK) cell activity (below the 90% tolerance limit) was identified in replicate experiments and 60% of them were young women (ages 20--39) . In vitro boosting of NK activity with Bacillus Calmette-Guerin (BCG) was also studied; overall, 56% of normal individuals responded positively to BCG . There was a significant (P less than 0.001) correlation between the unstimulated level of NCMC and the in vitro boosting with BCG, as 63% of individuals with a normal level of NK activity could be boosted as against only 19% of persons with low NK activity . We have also established the in vivo relevance of this in vitro test by determining the degree of correlation between responses to in vitro boosting with BCG and a positive or negative reaction in a hypersensitivity skin test using 5 IU of PPD (purified protein derivative of BCG) . Our results indicate that NCMC is an individual trait that varies little under physiological conditions, and that the response to BCG is a characteristic property of the effector lymphocyte, depending primarily on the unstimulated level of NCMC. Rev Fr Mal Respir, 1983, 11(5), 699 - 704 {Resistance of the Koch bacillus to antibacillary agents in the Department of the Lower Rhine}; Cooreman J et al.; The aim of this work was to study drug resistance of M . tuberculosis to Streptomycin (SM), Isoniazid (INH), Ethambutol (EMB) and Rifampicin (RMP) in the department of the Bas-Rhin from 1971 to 1978 . On 2,995 original culture plates which were positive, 1,561 antibiograms were performed at the C.E.R.P . The proportion of those tested has increased over the years from 27.8% in 1971 to 80.6% in 1978 because of the increasing number of laboratories participating in the study . The research is based on 1,511 cultures of M . tuberculosis and the population was split into two groups: 1,386 untreated subjects or treated for less than 15 days (primary resistance) and 125 subjects who had already been treated (acquired resistance) . Over all the years, primary resistance to at least one drug was 6.9% . Primary resistance to SM was 3.0%, to INH 2.2% and to both 1.4% (the levels of primary resistance to EMB and RMP were practically nil) . Acquired resistance to at least one of these antibiotics was 16.0% with the same order as for primary resistance: resistance to SM alone = 7.2%, to INH alone 4.0% and both together 2.4% . The proportions were greater than for primary resistance but concerned fewer subjects as only 8% of the population had been previously treated . There were no cases of primary or acquired resistance to Rifampicin alone. Mol Gen Genet, 1983, 192(3), 335 - 41 Molecular cloning of the genes for xylan degradation of Bacillus pumilus and their expression in Escherichia coli; Panbangred W et al.; The 7.7 Mdal PstI fragment of Bacillus pumilus IPO containing genes for xylan degradation, xylanase, and beta-xylosidase was inserted at the PstI site of pBR322 and cloned in E . coli C600 . The hybrid plasmid thus formed was named pOXN29 . The amount of xylanase and beta-xylosidase expressed in E . coli harboring pOXN29 was about 6% and 20% of the activity produced by the donor, B . pumilus . The reverse orientation of the inserted fragment resulted respectively in 5 times and 50 times increases in xylanase and beta-xylosidase productivities . Both enzymes expressed in E . coli transformants were shown to be indistinguishable from those of B . pumilus by immunological and chemical criteria . Digestion of pOXN29 with BglII produced two fragments; one was 6.7 Mdal in size and contained the whole pBR322 and the beta-xylosidase gene, and the other was 3.7 Mdal and coded for xylanase . Analysis of enzymes expressed in the transformant cells indicated that neither enzyme was secreted into the culture medium, periplasm nor membrane bound, although xylanase but not beta-xylosidase, was secreted into the medium in a B . pumilus culture. Mol Cell Biochem, 1983, 53-54(1-2), 155 - 61 Nonsubstrate induction of a soluble bacterial cytochrome P-450 monooxygenase by phenobarbital and its analogs; Fulco AJ et al.; A soluble, cytochrome P-450-dependent fatty acid hydroxylase--epoxidase complex from Bacillus megaterium ATCC 14581 can be induced more than 100-fold by the addition of phenobarbital or one of its analogs (hexobarbital) to the growth medium . These barbiturate inducers are apparently not substrates for the enzyme nor do they activate the monooxygenase in the cell-free system . The induction efficiency of both phenobarbital and hexobarbital can be significantly increased with respect to monooxygenase activity by autoclaving the inducer in the growth medium rather than by adding it to the medium after autoclaving . Turnover numbers of about 3 000 nmoles of substrate oxygenated per min per nmole of P-450 were obtained in crude cell-free preparations obtained from maximally induced cultures . Our data indicate that products formed by heating phenobarbital or hexobarbital in the growth medium are significantly better inducers of monooxygenase activity than are the unaltered drugs. Ann Nutr Metab, 1983, 27(4), 286 - 95 {Effect of moderate protein-calorie deprivation on cell-mediated immunity in mice}; Conge GA et al.; Mild protein-calorie deprivation in mice decreases their early nonspecific inflammatory response to an injection of BCG or tuberculin into the footpad . After an injection of BCG, the size of the granuloma and the delayed hypersensitivity reaction to tuberculin significantly decreased . This decrease in cell-mediated immunity was paradoxically accompanied by a fall in bacillus multiplication at the popliteal ganglion, i.e . the site of BCG injection . A reduction in microbial multiplication was also observed in the spleen after intraperitoneal injection of Listeria and in the liver after intravenous injection. Microbios, 1983, 36(144), 113 - 25 Ampicillin induced septum formation in Bacillus cereus; Salt WG et al.; Cells of Bacillus cereus grown in the presence of subinhibitory concentrations of ampicillin at either 30 degrees or 45 degrees C exhibited an increase in the numbers of centres of septum formation per unit cell length . Under identical conditions of cultivation, cells of Escherichia coli grew as aseptate filaments . In general, untreated B . cereus cells grown at 45 degrees C were longer than those grown at 30 degrees C . The strain of E . coli used was unaffected in terms of filamentation by elevated growth temperature . Results are discussed in terms of the presence and availability of penicillin binding proteins and autolysins involved in cell growth, division and separation. Folia Microbiol (Praha), 1983, 28(2), 65 - 70 Suppression of an exocellular proteinase synthesis in Bacillus megaterium by increased temperature; Vavrova M et al.; During cultivation of Bacillus megaterium at 42 degrees C the amount of the exocellular protease produced by growing cells sharply decreases as compared with temperatures of 28 and 35 degrees C . Within the above range the growth rate and incorporation of amino acids increase with increasing temperature . The culture adapted to 42 degrees C does not produce more proteinase at this temperature than the non-adapted culture . The high temperature does not induce accumulation of the enzyme in the cells . Total protein excretion was slightly lower at 42 degrees C than at 28 and 35 degrees C. Mikrobiologiia, 1983 Jan-Feb, 52(1), 33 - 8 {Characteristics of the autoregulatory factor d2 causing autolysis of Pseudomonas carboxydoflava and Bacillus cereus cells}; Svetlichnyi VA et al.; In the course of organoheterotrophous growth, Pseudomonas carboxydoflava Z-1107 and Bacillus cereus 504 were found to synthesize the autoregulatory factor d2, a membranotropic lipid metabolite, and to accumulate it in the growth medium . At a low concentration, the factor activated respiration of the cells: at a high concentration, it inhibited respiration and induced autolysis of the cells . The physiological action of d2 was shown to be due to the effect of free fatty acids, in particular, palmitic, stearic and oleic acids . Oleic acid displayed the highest physiological activity, P . carboxydoflava Z-1107 became more susceptible to high concentrations of d2 and oleic acid, and respiration of the cells was activated by low concentration of these compounds when the culture passed from the exponential growth phase to the linear one . These results as well as data reported in literature about the action of fatty acids on biological membrane suggest that low concentrations of the factor d2 uncouple respiration and oxidative phosphorylation whereas high concentrations of the factor disorganize the structure of the cytoplasmic membrane by increasing its fluidity. Folia Microbiol (Praha), 1983, 28(1), 46 - 50 Combined effect of temperature and nutrients on protein turnover in Bacillus megaterium; Prasad R et al.; Protein turnover was followed in populations of Bacillus megaterium growing in temperature range of 17-48 degrees C in different media . Higher temperature stimulated the protein turnover (expressed as the amount of protein degraded during 3.3 h) in all the media tested up to the optimal growth temperature (40-42 degrees C) . Protein turnover in a medium containing amino acids continued to be stimulated by temperature even above this point; no further significant increase of turnover was found in the other media. J Urol, 1983 Jan, 129(1), 33 - 4 A comparison of treatment methods for the prophylaxis of recurrent superficial bladder tumors; Rodrigues Netto Junior N et al.; We assigned 55 patients with superficial transitional cell carcinoma of the bladder by rotation to 1 of 3 treatment categories . Group 1 patients were treated by transurethral resection alone, group 2 by transurethral resection followed by topical thiotepa and group 3 by transurethral resection followed by orally administered bacillus Calmette-Guerin . Recurrent bladder tumors developed subsequently in 80 per cent of the patients in group 1, 42.9 per cent in group 2 and 6.2 per cent in group 3. J Bacteriol, 1983 Jan, 153(1), 475 - 84 Purification and characterization of a Bacillus megaterium disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate; Swerdlow RD et al.; An NADH-linked disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate moieties was purified 23,000-fold to homogeneity from spores of Bacillus megaterium . The enzyme had a native molecular weight of 122,000 with two apparently identical subunits, contained one molecule of flavin adenine dinucleotide per subunit, and was inhibited by the vicinal dithiol reagent arsenite . The enzyme was active only on disulfides containing pantethine 4',4"-diphosphate moieties, including pantethine 4',4"-diphosphate, oxidized coenzyme A, and coenzyme A in disulfide linkage to acyl carrier protein . However, the Km values for pantethine 4',4"-diphosphate and oxidized coenzyme A were 0.65 and 7.4 mM, respectively . The enzyme was at a low level in log-phase cells but increased up to 10-fold early in the stationary phase and had a similar specific activity in both the mother cell and the forespore compartment; the enzyme activity fell only slowly during spore germination and outgrowth . The enzyme was not detected in several eucaryotic sources and was present in at most a low level in a number of gram-negative bacteria . Surprisingly, the specific activity of this enzyme varied more than 200-fold in extracts from different Bacillus species, with values in B . subtilis being 5- to 6-fold lower and values in B . cereus and B . sphaericus being 8- and 35-fold higher, respectively, than the maximum value in B . megaterium . However, the high specific activity in B . sphaericus did not represent more enzyme protein than in B . megaterium . The possible function of this newly discovered enzyme is discussed. J Bacteriol, 1983 Jan, 153(1), 436 - 42 Dielectric characterization of forespores isolated from Bacillus megaterium ATCC 19213; Marquis RE et al.; Isolated stage III forespores of Bacillus megaterium ATCC 19213 in aqueous suspensions were nearly as dehydrated as mature spores, as indicated by low dextran-impermeable volumes of ca . 3.0 ml per g (dry weight) of cells compared with values of ca . 2.6 for mature spores and 7.3 for vegetative cells . The forespores lacked dipicolinate, had only minimal levels of calcium, magnesium, manganese, potassium, and sodium, and were more heat sensitive than vegetative cells . The effective homogeneous conductivities and dielectric constants measured over a frequency range of 1 to 200 MHz indicated that the inherent conductivities of the forespores were unusually low, in keeping with their low mineral contents, but that the forespores could be invaded by environmental ions which could penetrate dielectrically effective membranes . Overall, our findings support the view that the dehydration of a forespore during stage III of sporogenesis may be the result of ion movements out of the forespore into the sporangium. J Bacteriol, 1983 Jan, 153(1), 375 - 8 Enzymatic activity of precursors of Bacillus megaterium spore protease; Hackett RH et al.; The protease that initiates rapid proteolysis during germination of Bacillus megaterium spores is synthesized during sporulation as a 46,000-molecular-weight polypeptide (P46) and is processed later in sporulation to a 41,000-molecular-weight polypeptide (P41), which is converted to a 40,000-molecular-weight polypeptide (P40) early in spore germination . P40 is known to be both tetrameric and enzymatically active . In this work, we show that P46 and P41 are both tetrameric, but that only P41 is enzymatically active . The identification of a zymogen form (P46) of this protease explains in part the regulation of the activity of this enzyme. Ann N Y Acad Sci, 1983, 420, 237 - 50 Detection of human malignant melanoma antigens by immunofluorescence and autologous postimmune antimelanoma sera; Leong SP; The objectives of this study are to demonstrate the specificity of sera from melanoma patients undergoing autologous immunization and to localize melanoma antigens on a cellular level . Five melanoma patients were immunized with autologous melanoma cells and Bacillus Calmette-Guerin . This immunization program was conducted at Tulane University . Indirect immunofluorescence using both viable and fixed melanoma cells was employed . Four of five postimmune sera were reactive to five of seven melanoma cell lines . Two of the four reactive antisera showed positive binding with two additional melanoma lines obtained from other laboratories . All these sera were negative against seven nonmelanoma lines . Negative controls consisted of sera from 65 nonimmunized melanoma patients and 140 nonmelanoma patients . Membrane immunofluorescence (MIF) demonstrated sequential full MIF, capping, polarization, and extrusion of antigen-antibody complexes on the cell surface . MIF inhibition showed shedding of melanoma antigens in the culture medium . Ethanol, methanol, formalin, trichloroacetic acid, and acetone yielded sharp MIF . Isopentane and isooctane gave bright cytoplasmic fluorescence . In conclusion, this study provides suggestive evidence for the existence of common melanoma antigens as defined by the postimmune antimelanoma sera . These antigens may be localized in the membrane or within the cytoplasm. Probl Med Wieku Rozwoj, 1983, 12, 363 - 75 {High-molecular-weight lipoprotein concentrations in pneumonia in children and experimental animals}; Kaminska M et al.; Bacterial sepsis can alter the level of serum lipids in man . We have been to find any reports on lipoproteins in children with pneumonia . The study group comprised 25 children from 9 months to 4 years suffering from severe pneumonia . The initial phase of disease in most of the children involved an increase in endogenous triglyceride values and a decrease in the lipids of high density lipoproteins (HDL) . An increase in VLDL and decrease in VLDL and decrease in HDL were confirmed by lipoprotein electrophoresis . The drastic fall in HDL resulted in HDL-cholesterol and phospholipid levels as low as 50 and 100 mg/l respectively . Etiological factors were gramnegative bacteria . On remission HDL levels increased and that of VLDL decreased . Similar lipid investigations have been performed in animals (rabbits) . The results of these investigations were convergent with those obtained in children . The extend and intensity of the lung changes were not, however, connected with the kind of bacillus with which the children were infected . It is probable that retention of triglycerides is the result of the reduction in HDL concentration . The question arises whether the low HDL level can be linked with synthesis of surfactant lecithin in pulmonary alveoli, or with endotoxin. Breast Cancer Res Treat, 1983, 3 Suppl, S61 - 8 Adjuvant endocrine therapy, cytotoxic chemotherapy, and immunotherapy in stage-II breast cancer: five-year results; Pearson OH et al.; Five-year results of a prospective, randomized clinical trial of three treatment regimes--(a) cytoxan, methotrexate, and 5-fluorouracil (CMF); (b) CMF plus the antiestrogen drug, tamoxifen (CMFT); and (c) CMFT plus bacillus Calmette-Guerin (BCG) vaccinations--in 312 women with stage-II breast cancer are reported . Estrogen receptors (ER) were measured in all of the primary tumors . Addition of tamoxifen to CMF therapy significantly decreased the number of recurrences at five years in ER positive patients with four or more positive axillary lymph nodes . Addition of tamoxifen to CMF had no effect on disease-free survival in ER-positive patients with 1-3 positive axillary lymph nodes or in patients with ER-negative tumors . Addition of BCG vaccinations had no discernible effect on disease-free survival . ER measurements in the primary tumor provide important prognostic information regardless of treatment, with ER-positive patients having lower recurrence rates and mortality after five years . ER measurements also have predictive value for response to endocrine therapy . Further follow-up is needed to determine whether tamoxifen is delaying recurrence or preventing it in a subset of these patients. Z Allg Mikrobiol, 1983, 23(8), 517 - 35 {Molecular biology of the germination of Bacillus spores}; Hecker M; The review deals with recent results and problems of gene expression during germination of Bacillus spores . Three problems were selected: 1 . The activation of metabolism as a prerequisite for the synthesis of nucleic acids and proteins . 2 . The activation of nucleic acid and protein synthesis during germination . 3 . The gene expression programme of germinating spores . Using the highly sensitive two-dimensional polyacrylamide gel analysis three major classes of proteins were distinguished, depending on the time of onset and duration of their syntheses: a) proteins made throughout germination (main class), b) proteins whose synthesis started only after a lag phase and then continued throughout germination, and c) proteins which are synthesized only during the early phases of germination . The programme of protein synthesis is an indicator for the control of gene expression during germination . The regulation of expression of these major gene groups during spore outgrowth is discussed. Chemotherapy, 1983, 29(6), 428 - 35 Efficacy of trimethoprim/sulfamethoxazole in experimental Escherichia coli bacteremia and meningitis; Kim KS et al.; We evaluated the activity of trimethoprim/sulfamethoxazole (TMP/SMZ) against a K1 Escherichia coli strain . Minimal inhibitory and bactericidal concentrations were 0.06/1.14 and 0.25/4.75 micrograms/ml, respectively . In vivo studies using an infant rat model of bacteremia and meningitis revealed that TMP/SMZ penetrated well into the cerebrospinal fluid (CSF) and that 37% of serum levels were achieved . The efficacy of TMP/SMZ was compared with that of ampicillin, chloramphenicol, cefotaxime and lamoxactam . Bacterial clearance from blood and CSF was significantly greater with TMP/SMZ than with ampicillin or chloramphenicol and mortality was significantly less than with chloramphenicol (p less than 0.01) . However, 3 of 21 (14%) and 2 of 8 animals (25%) still had positive blood and CSF cultures after 3 days of treatment with TMP/SMZ . None of the survivors in the cefotaxime and lamoxactam groups were bacteremic after 1 day of therapy . Furthermore, 5 of 13 animals (38%) treated with TMP/SMZ developed meningitis during therapy, in contrast with none in the cefotaxime and lamoxactam groups . These findings indicate that although the activity of TMP/SMZ is bactericidal in vitro and in vivo against E . coli, TMP/SMZ may not provide optimal therapy for gram-negative bacillary meningitis in this model. J Biol Response Mod, 1983, 2(4), 321 - 42 Clinical immunotherapy trials of bacterial components derived from Mycobacteria and Nocardia; Vosika GJ; Preparations of oil-attached mycobacterial components have been used in place of viable bacille Calmette-Guerin (BCG) in animal models and humans as a cancer immunotherapeutic agent . Most preparations consist of the isolated mycobacterial cell wall or cell wall skeleton attached to oil . Cord factor (trehalose dimycolate) has also been included in some preparations . In animal models, such preparations given intralesionally, systemically, or as a vaccine can cause regression of disease and establish tumor-specific immunity . Trials in humans have utilized oil-attached mycobacterial components given intralesionally, intradermally, intrapleurally, intraperitoneally, intravenously, and as an ointment . The major toxicity has been fever, chills, and local inflammation and/or abscess formation . An increase in the white blood count and lymphocyte count has been observed . An increase in liver function test was reported in a minority of patients . Given intralesionally, these preparations cause regression of the injected lesion, regression of noninjected cutaneous and visceral disease, and the apparent establishment of a tumor-specific immune response . Administered intrapleurally and intraperitoneally, there is a response and a clearing of malignant cells in approximately 50% of cases . Given intravenously, oil-attached cell wall skeleton and trehalose dimycolate can eradicate pulmonary disease . Used as an ointment, the preparations have been effective in mycosis fungoides and Kaposi's sarcoma . These reagents demonstrate definite single-agent activity . This was most prominent in patients who were immunocompetent and who had immunogenic tumors such as malignant melanomas . The reagents represent potent immunotherapeutic agents with acceptable toxicity . Further trials of these and subsequent refined preparations are warranted. Z Erkr Atmungsorgane, 1983, 161(1), 56 - 60 {Treatment of tuberculosis before the introduction of chemotherapy}; Sighart H; 20 years before the detection of the tubercle bacillus by Robert Koch Brehmer and Dettweiler have created the fundamentals of tuberculosis therapy . The rest-cure in the open air initiated by them and the system of sanatorium treatment resulting from it have formed the management of tuberculosis all over the world for nearly 100 years . Even collapse therapy in the heroic phase of tuberculosis treatment has remained only a supplement to conservative therapy . In the centre of the scientific discussions, substantially influenced also by the "Wiener Schule", there were the problems of the first manifestations of tuberculous disease its early detection, giving the best chances of cure at those times; there was the knowledge that the cavity was of crucial significance for the fate of the tuberculous patient; in consequence thoracic collapse surgery was developed. Med Microbiol Immunol (Berl), 1983, 172(1), 23 - 32 Bacteria-immune system interactions . V . Escherichia coli cell-wall components involved in binding to lymphocytes; Shockley RK et al.; Various bacterial cell-wall components were tested for their ability to inhibit Escherichia coli 2 binding to lymphocytes . The binding was inhibited by outer membrane extracts of E . coli 2 but not by similar extracts from E . coli 0 . In addition, mannose-binding protein, inner membrane or lipopolysaccharide extracts of E . coli 2 or lipoteichoic acid extracts of Bacillus globigii had no effect on lymphocyte-E . coli 2 interaction . The interaction did not appear to be mediated by fimbriae, since such structures could not be detected by electron microscopy . Comparison of the outer membrane proteins of a binder (E . coli 2) and a nonbinder (E . coli 0) bacteria revealed differences in their number and position. Ann Immunol (Paris), 1983 Jan-Feb, 134C(1), 125 - 47 Viability, heat stability and immunogenicity of four BCG vaccines prepared from four different BCG strains; Gheorghiu M et al.; Four BCG strains, French 1173-P2, Danish 1331, Japanese 172 and Glaxo-1077, among those most widely used throughout the world for tuberculosis vaccination, were prepared according to the respective conditions in each BCG-production unit . These 16 vaccine preparations were distributed and compared using standardized in vitro and in vivo conditions, based on optical density and bacillary mass/ml . The results presented in this report concern only the 4 freeze-dried BCG strains prepared by the Institut Pasteur BCG unit . It is shown that these 4 preparations differ in their in vitro characteristics such as growth, morphology, heat stability and viability, tested by colony-forming units and the ATP content . In addition, statistically significant differences were observed in biological control tests performed in guinea-pigs and mice when they were immunized with the same concentrations of 4 reconstituted vaccines . For instance, in tests measuring hallmarks of cell-mediated immunity--such as local granuloma formation, spleen index, delayed-type hypersensitivity to tuberculin, and non-specific immunopotentiation to unrelated antigen--a ranking order of relative immunopotency of the four strains was made, giving the following classification in decreasing order: French 1173-P2, Danish 1331, Japanese 172 and Glaxo-1077 . Specific acquired resistance after BCG vaccination in mice was obtained with all four vaccines . However, higher protection was observed in mice vaccinated with Glaxo-1077 and French 1173-P2 strains . No obvious correlation existed between the intensity of cellular post-vaccination resistance and the above CMI tests, except those concerning the three-week index of BCG infection measured in the spleen, which correlated with the protection test. Scand J Immunol, 1983 Jan, 17(1), 37 - 43 Breakdown product of factor B as an index of complement activation in lepromatous leprosy and its relation with bacillary load; Saha K et al.; Serum complement profile studied in 50 lepromatous leprosy patients with various bacillary loads demonstrated significantly decreased C3 levels in patients with high bacteriological index (2+ to 4+) as compared with those with lesser bacterial load . In contrast, mean serum levels of C1q and C4 components remained unchanged . The concentration of factor B breakdown product (Ba) and its ratio to factor B increased with the bacterial density and more so in patients with erythema nodosum leprosum . A significant negative correlation was found between serum C3 and Ba levels in most lepromatous patients . Analysis of the data suggested the alternative pathway as the possible mechanism of complement activation in lepromatous leprosy. Mikrobiologiia, 1983 Jan-Feb, 52(1), 46 - 9 {Activity of intracellular enzymes in Bacillus thuringiensis prospores and sporangia}; Krainova OA et al.; The activity of endocellular enzymes (alkaline phosphatase, protease, glucose dehydrogenase, aldolase, malate dehydrogenase, NADH dehydrogenase, NADH oxidase) was studied in isolated prospores and sporangia as well as in vegetative cells of Bacillus thuringiensis strains, one of which produced crystals and one did not . The activity of malate dehydrogenase and NADH dehydrogenase was high in prospores of the both strains at the fifth and sixth stages of spore formation . The strain which did not produce crystals differed from the parent strain by a higher aldolase activity at all of the growth stages and by an abrupt increase in the activity of hydrolytic enzymes in sporangia (in the cytoplasm of the parent cells). Cancer Immunol Immunother, 1983, 14(3), 167 - 71 Postoperative adjuvant immunotherapy of gastric cancer with BCG-cell wall skeleton . 3- to 6-year follow up of a randomized clinical trial; Ochiai T et al.; The clinical effectiveness of immunotherapy with the cell wall skeleton of Bacillus Calmette-Guerin was assessed in a study involving 140 consecutive patients with gastric cancer, who were gastrectomized at a single institution from January 1976 through December 1978 . These patients were randomized by an envelope method after operation and divided into three treatment groups: 'control', 'chemotherapy', and 'chemotherapy plus immunotherapy with BCG-CWS.' Only two patients who died during surgery were excluded, and a survey of survival periods was made on the remaining 138 patients in January 1982 . As a result, statistically significant differences in the survival rate curve were observed between the control and chemotherapy plus immunotherapy groups (P less than 0.01), and between the chemotherapy and chemotherapy plus immunotherapy groups (P less than 0.05) . These results emphasize effectiveness of BCG-CWS as an adjuvant immunotherapeutic agent in gastrectomized cancer patients. Cancer Immunol Immunother, 1983, 14(3), 151 - 4 Eradication of microscopic hepatic metastases by active specific immunization; Sukumar S et al.; Strain-2 guinea pigs, each with microscopic deposits of line 10 hepatocarcinoma in the liver, were treated by ID immunization with a mixture of irradiated tumor cells and an oil-in-water emulsion containing cell walls of Mycobacterium bovis strain Bacillus Calmette-Guerin (BCG CWE) . Injection of line 10 hepatoma cells into the hepatic portal vein led to the development of tumor foci in the liver, metastasis in the hepatic lymph node, malignant ascites, and death . Active immunization using irradiated line 10 cells and BCG CWE was effective therapy when administered 1, 7, and 14 days after intraportal injection of line 10 cells . Effective immunization required both irradiated line 10 tumor cells and the BCG cell wall emulsion . Immunization with BCG CWE admixed with irradiated line 1 tumor cells, a hepatoma antigenically distinct from line 10, did not prevent outgrowth of line 10 deposits in the liver . Animals rendered free of disease could reject a challenge of line 10 tumor cells but not of line 1 tumor cells. Oncology, 1983, 40(2), 95 - 101 Serological detection of common human melanoma membrane antigens by microcomplement fixation and immunofluorescence; Leong SP et al.; An antiserum to human melanoma antigens was obtained from a melanoma patient after immunization with autologous irradiated cultured melanoma cells and bacillus Calmette-Guerin . Using the microcomplement fixation assay, the antiserum, at a titer of 1/1,800, was noted to bind strongly with 7 of 10 allogeneic cultured human melanoma cells lines . However, using the indirect immunofluorescence test and serum at a much lower titer (1/8), only 3 of the 10 melanoma cell lines were positive . Using both microcomplement fixation and indirect immunofluorescence, no significant reactivity was noted in several nonmelanoma cell lines including Hela, human lung adenocarcinoma, human prostatic carcinoma, WI-38 and VA-13 cell lines . These data suggest that common melanoma membrane antigens exist on 7 of 10 cultured human melanoma cell lines as tested by microcomplement fixation and that this assay is more sensitive than immunofluorescence . These common melanoma membrane antigens may eventually be extracted, purified and used for specific immunodiagnosis and immunotherapy. Int Arch Allergy Appl Immunol, 1983, 70(1), 65 - 70 Tuberculin anergy in clinically normal individuals . I . Lymphokine and lymphocyte transformation studies; Muller HK et al.; Tuberculin anergy was demonstrated in a number of clinically normal individuals who, on being immunized with Bacillus Calmette-Guerin (BCG), failed to demonstrate a positive Mantoux skin test . Lymphoid cells from these anergic subjects when stimulated with purified protein derivative (PPD) in vitro did not produce macrophage migration inhibition factor (MIF) . Lymphocyte transformation studies demonstrated that while Mantoux-positive, non-BCG immunized Mantoux-negative and anergic individuals could all undergo blast cell transformation in the presence of PHA, only lymphoid cells from Mantoux-positive and a proportion of anergic people were capable of blast cell transformation when cultured with PPD . These results suggest that more than one mechanism may account for tuberculin anergy in clinically normal people operating at the level of lymphokine production and in some cases lymphocyte transformation. Z Allg Mikrobiol, 1983, 23(5), 313 - 7 {Biological effects of coordination compounds of transitional metals . The beta-lactamase inhibitory effect of cis-platin, 2-aminopyridine palladium chloride, clavulanic acid and penicillanic acid sulfonate CP 45,899 in the nitrocefin test and Titertek/microtiter automatic system}; Fleck WF et al.; The beta-lactamase-inhibiting activity of 6m-ethyl-pyrid-2-yl-ammine palladium-dichloride (Pd 25681) and cis-dichloro-diammine-platinum(II) was studied and compared with the enzyme inhibitory action of potassium clavulanate and the penicillanic acid sulfone CP 45899 . Using the nitrocefin test method and the Titertek/Microtiter equipment CP 45899 and potassium clavulanate were the strongest inhibitors of the Bacillus cereus beta-lactamase . Cis-dichloro-diammine-platinum(II) was fourfold less active than the palladium complex PD 25681 in aquimolar concentration . The following ID50 values were found: CP 45899: 0.0281 microgram; K-clavulanate: 0.1274 microgram; Pd 25681: 3.8603 microgram; cis-dichlorodiammine-platinum(II): 12.5120 microgram/100 microliter. Transplantation, 1983 Jan, 35(1), 56 - 61 Mechanisms of immunological eradication of a syngeneic guinea pig tumor . II . Effect of methotrexate treatment and T cell depletion of the recipient on adoptive immunity; Shu S et al.; The influence of methotrexate on the development of immunity to the line 10 hepatoma was studied in guinea pigs . Chronic methotrexate treatment had no apparent effect on the ability of immune guinea pigs to suppress the growth of inoculated tumor cells . In contrast, the same methotrexate regimen inhibited the development of tumor immunity if started before the 8th day after immunization with a vaccine containing viable line 10 cells admixed with Bacillus Calmette-Guerin (BCG) cell walls . Thus, methotrexate selectively inhibited the afferent limb of the immune response . In adoptive transfer experiments, methotrexate-treated recipient guinea pigs were capable of being passively sensitized with immune spleen cells, indicating that the primary cell-mediated immune response of the recipient was not required for adoptive immunity . The contribution of recipient T cells in adoptive immunity was further investigated in guinea pigs deleted of T cells by thymectomy, irradiation, and bone marrow reconstitution . Despite demonstrable deficiency in T lymphocyte reactions, "B" animals were fully capable of rejecting tumors after transfer of immune cells . These results suggest that the expression of adoptive immunity was independent of recipient T cell participation . In addition, sublethal irradiation of immune spleen cells prior to adoptive transfer abolished their efficacy . Proliferation of transferred immune cells in the recipient may be essential for expression of adoptive immunity. J Immunopharmacol, 1983, 5(3), 173 - 90 Bacteria-immune system interactions . VIII . Modulation of human cytotoxic reactions by crude lipoteichoic acid extracts of Bacillus globigii; Kleinman R et al.; Crude lipoteichoic acid (LTA)1 extracts were prepared from Bacillus globigii (Bg-LTA) . The addition of Bg-LTA to mixed lymphocyte reactions (MLR) produced a dose dependent inhibition of specific cytotoxic reactions and of tritiated thymidine (3H-TdR) incorporation . Much less Bg-LTA was needed to inhibit the initiation (addition at day 0) than the effector phase (addition at day 6) of the cytotoxic reactions . None of the LTA quantities used had a toxic effect on the lymphocytes . LTA pretreated stimulator cells also had a modulatory effect on the cytotoxic reactions . Stimulators pretreated with very low or very high LTA concentrations had no effect; however, pretreatment with intermediate concentrations induced a drastic inhibition of cytotoxicity . LTA pretreated stimulator cells did not significantly affect 3H-TdR incorporation . Suppression of the cytotoxic reaction could also be obtained by LTA-pretreatment of the effector cells. J Interferon Res, 1983, 3(1), 129 - 37 In vitro production of gamma interferon is dependent on the mouse genotype; Huygen K et al.; C57B1/6 and BALB/c mice were compared for their in vitro capacity to produce gamma interferon (IFN-gamma) in response to specific antigen (i.e., Purified Protein Derivative in bacillus Calmette-Guerin sensitized animals) and T-cell mitogen (Concanavalin A) . In both cases C57B1/6 spleen cells produced five to ten times more IFN than identically treated BALB/c spleen cells . More detailed analysis of in vitro induced BCG-PPD IFN indicates that the low IFN level in BALB/c cultures is neither caused by classical splenic suppressor cells such as macrophages or T-cells, nor by inactivation of secreted IFN by inhibitory substances. Acta Microbiol Pol, 1983, 32(3), 269 - 74 The quality of protein from algae used in the purification of wastewater from the nitrogen fertilizer industry; Miernik A; This paper presents the quantities and amino acid composition of proteins from Chlorella vulgaris, Scenedesmus obliquus and Stichococcus bacillaris used in purification experiments on wastewater from the Nitrogen Fertilizer Factory in Chorzow . The quantities of exogenous amino acids of the investigated algae and "ideal protein" standard of the FAO are compared . The protein from Scenedesmus and Chlorella appeared to be richer in exogenous amino acids than the FAO standard . Stichococcus bacillaris has a lower content of exogenous amino acids but nevertheless has a high content of threonine and lysine, higher than in the other investigated algae. Mikrobiologiia, 1983 Jan-Feb, 52(1), 87 - 93 {Use of rifampicin for directed modification of the nucleotide composition of the cells of Bacillus megaterium and Candida utilis}; Rabotnova IL et al.; The object of this work was to study the effect of rifampicin on the physiological state of Bacillus megaterium and Candida utilis in chemostat and batch cultures . When B . megaterium is cultivated in a medium containing 1 microgram of rifampicin per litre, its metabolism changes, the pool of free nucleotides rises, and the content of RNA drops in response to the specific action of the inhibitor . The modified state is observed throughout many generations . The action of the inhibitor is also non-specific: protein content falls and polyhydroxybutyric acid is accumulated . The morphology of B . megaterium cells changes: they become larger, their cell wall thickens, and the separation of cells from one another is complicated . At a high concentration (5 mg/l), rifampicin decelerates the growth of C . utilis and causes merely a minor non-specific inhibition. J Biochem (Tokyo), 1983 Jan, 93(1), 177 - 88 Transfer of D-phenylalanine from gramicidin S synthetase 1 to gramicidin S synthetase 2 in gramicidin S synthesis; Hori K et al.; The transfer of phenylalanine from gramicidin S synthetase 1 (GS 1) to gramicidin S synthetase 2 (GS 2) was studied by the use of combinations of wild-type GS 1 with various GS 2s from a wild strain and gramicidin S non-producing mutant strains of Bacillus brevis Nagano . The combinations of mutant GS 2s lacking 4'-phosphopantetheine (from BI-4, C-3, E-1, and E-2) did not transfer D-phenylalanine from GS 1, although they could activate all the constituent amino acids . Other mutant GS 2s containing 4'-phosphopantetheine, except GS 2 from BII-3 (proline-activation lacking) accepted D-phenylalanine from intact GS 1 . To ascertain more directly whether 4'-phosphopantetheine is involved in the transfer of D-phenylalanine from GS 1 to GS 2, pepsin digests of GS 2 that accepted {14C}phenylalanine were analyzed by Sephadex G-50 column chromatography and thin-layer chromatography (TLC) . Radioactivity of {14C}phenylalanine was always associated with a peptide containing 4'-phosphopantetheine . Furthermore, the position of radioactivity was distinct from the position of 4'-phosphopantetheine on TLC after alkaline treatment or performic acid oxidation of the digests. J Hand Surg {Am}, 1983 Jan, 8(1), 39 - 42 Toxic lesions of the hand associated with chemotherapy; Seyfer AE et al.; New agents used in cancer chemotherapy are continually appearing which, in turn, leads to newer patterns of toxicity . This report reviews experience with some upper extremity lesions associated with chemotherapy . Doxorubicin extravasation caused deep soft tissue necrosis that was successfully managed by aggressive excision and delayed coverage with a well-vascularized flap . Bleomycin resulted in palmar swelling, erythema, and digital pulp necrosis, which healed secondarily . Intravenous nitrogen mustard caused an immediate, and alarming, perivenous hyperpigmentation that was painless but long lasting . Bacille Calmette-Guerin immunotherapy was followed by a large, localized skin slough that was indolent, painful, and healed secondarily . 5-Fluorouracil was associated with permanent, dark pigmentation of the hands along with slough of the digital pulps . The myelosuppression resulting from combination therapy may also predispose these patients to fulminant hand infections, which can be lethal. Microbiol Immunol, 1983, 27(8), 695 - 708 Purification and characterization of beta-glucuronidase inhibitor from Mycobacterium tuberculosis; Kiyotani K et al.; Factors inhibitory to beta-glucuronidase were found in the culture filtrate and in a bacillary extract of Mycobacterium tuberculosis H37Rv grown for 6 weeks on Sauton medium . The inhibitors were purified by ammonium sulfate fractionation, treatment with n-butanol and streptomycin, and chromatography on DEAE-Sepharose CL-6B . Two inhibitors were obtained from the culture filtrate . The molecular weights were estimated to be 25,500 and 15,500 by gel filtration on a Sephadex G-75 column . Three inhibitors were purified from the bacillary extract, two of which were similar to those from the culture filtrate . The molecular weight of the third inhibitor was 21,000 . However, the molecular weight of all the denatured inhibitors was 8,600 in the presence of sodium dodecyl sulfate . The inhibitors contained extremely high amounts of glutamic and aspartic acids and had a highly acidic isoelectric point of pH 2.5 . The inhibitors acted noncompetitively against beta-glucuronidase of guinea pig origin at an optimal pH 4.5 . beta-Glucuronidases from human peripheral leukocytes and beef liver were partially sensitive to the inhibitors; all the other enzymes tested for sensitivity were unaffected by the inhibitors. Mol Gen Genet, 1983, 189(1), 181 - 3 Cloning and expression in Escherichia coli of a DNA fragment from Bacillus sphaericus coding for biocidal activity against mosquito larvae; Ganesan S et al.; A 3.7 kb DNA fragment from Bacillus sphaericus 1593 was cloned and expressed in E . coli using the plasmid pHV33 . The level of larvicidal activity of the hybrid plasmid, pGsp03, against two species of mosquito larvae was comparable to that found with sporulating cell suspensions of B . sphaericus . A limited restriction cleavage map of the cloned insert is given. Bioorg Khim, 1983 Jan, 9(1), 127 - 9 {Site-specific endonuclease BmeI from Bacillus megaterium 216}; Pachkunov DM et al.; A site-specific endonuclease BmeI has been isolated from Bacillus megaterium 216 by gel filtration on ultragel AcA-44 with a subsequent chromatography on heparin-sepharose 6B . On the double-stranded DNA the endonuclease recognizes the pentanucleotide sequence (Formula: see text); and hydrolyzes it in the points shown by arrows . At gel filtration the endonuclease is eluted in the volume corresponding to a molecular mass of 60 000. FEBS Lett, 1982 Dec 27, 150(2), 419 - 23 The tyrosyl-tRNA synthetase from Escherichia coli . Complete nucleotide sequence of the structural gene; Barker DG et al.; The structural component of the tyrS gene of Escherichia coli, comprising 1269 base pairs, has been fully sequenced by the combined M13/dideoxychain termination approach . The gene has a codon usage pattern which is typical of highly expressed proteins and similar to other Escherichia coli aminoacyl-tRNA synthetase genes . Peptide purification and sequencing has been used to locate the N-terminus and to provide confirmation of 95% of the translated protein sequence . This latter yields on Mr of 47,403 for the Escherichia coli tyrosyl-tRNA synthetase, and reveals considerable homology with the primary structure of the analogous enzyme isolated from Bacillus staerothermophilus. Eur J Biochem, 1982 Dec 15, 129(2), 403 - 7 Induction of sporulation in Bacillus brevis . 2 . Dependence on the presence of the peptide antibiotics tyrocidine and linear gramicidin; Pschorn W et al.; This paper presents evidence that the two peptide antibiotics tyrocidine and linear gramicidin, produced by Bacillus brevis ATCC 8185, are required for the induction of sporulation in the producer organism . When tyrocidine synthesis was specifically blocked with 2-amino-3-hydroxy-3-phenylpropanoic acid {Mach, B., Reich, E., and Tatum, E . L . (1963) Proc . Natl Acad . Sci . USA, 50, 175-181}, sporulation and gramicidin synthesis were inhibited, but both processes could be restored by the addition of tyrocidine . Certain other amino acids such as L-tyrosine inhibited both sporulation and peptide antibiotic synthesis in nitrogen-limited cultures . When either tyrocidine or linear gramicidin was added together with L-tyrosine, neither sporulation nor peptide antibiotic synthesis was restored . On the other hand, the addition of both tyrocidine and linear gramicidin effectively reversed the inhibition of sporulation by L-tyrosine . These experiments demonstrate that sporulation of B . brevis depends on either the endogenous synthesis or the addition of both tyrocidine and linear gramicidin . The fact that endogenous as well as exogenous peptides could effect sporulation argues against the involvement of artifacts, such as the depletion of intracellular nucleotide pools caused by the surfactant properties of added peptide antibiotics. Eur J Biochem, 1982 Dec 15, 129(2), 395 - 401 Induction of sporulation in Bacillus brevis . 1 . Biochemical events and modulation of RNA synthesis during induction by tyrocidine; Ristow H et al.; Under conditions of severe nitrogen starvation, brought about by nutritional shift-down, Bacillus brevis ATCC 8185 was unable to sporulate unless supplemented with the peptide antibiotic tyrocidine . The induction of sporulation was highly specific for tyrocidine and required only very low concentrations of the peptide (5 microM) . Tyrocidine-induced sporulation was accompanied by the typical sporulation-specific events (e.g . extracellular protease production and dipicolinate synthesis) as well as the formation of linear gramicidin . The addition of tyrocidine produced acute inhibition of RNA synthesis that was followed by a limited activation of transcription near the time of onset of linear gramicidin synthesis, when the first sporulation-specific changes were observed . These results provide direct evidence for a role of tyrocidine in sporulation of B . brevis and suggest that the action of the peptide antibiotic may involve the control of transcription . Such a notion is supported by earlier studies on the effects of tyrocidine and linear gramicidin on purified RNA polymerase. Biochim Biophys Acta, 1982 Dec 8, 693(1), 134 - 42 Hexagonal surface array in a protein-secreting bacterium, Bacillus brevis 47; Tsukagoshi N et al.; Bacillus brevis 47, a protein-secreting bacterium, contained two major proteins with approximate molecular weights of 150 000 and 130 000 in the cell wall . The cell surface was covered with a hexagonally arranged array of six structural units about 4 nm in diameter with a lattice constant of 14.5 nm . The regular array structure as well as the chemical composition of cell envelopes remained the same regardless of the growth conditions . A mutant, strain 47-57, which was isolated as a phage resistant colony, contained only the 150 000 protein as a major cell wall protein . Although the mutant had hexagonally arranged arrays with the same lattice constant as that of wild-type cells, the distribution of mass in the unit cell differed considerably from that of the wild-type cells . The number of structural units in the unit cell of the mutant was reduced from six to three . Taking these results together with filtered images of the wild-type and mutant envelopes, two possible models for the surface array of B . brevis 47 are discussed. C R Seances Acad Sci III, 1982 Dec 6, 295(12), 679 - 82 {Enzymatic release of sedimentary bacteria in the presence of antibiotics}; Brisou JF et al.; Polysaccharases release microorganisms from their natural seat, marine sediments for example . The enzymatic activity works both on the microbial adherence polysaccharides and on the support surfaces (cellulose, pectine, etc.) . Dosages of glucose confirm polysaccharase activity . An association of bacitracine, thiophenicol and a few enzymes: cellulase, pectinase, amyloglucosidase, alpha amylase, hyaluronidase, release a considerable number of bacteria . The culture on specific mediums confirm the specificity of this release . E . coli polyresistant strain where isolated by amylo-glucosidase, glucuronidase association in a mixture of thiophenicol and bacitracine . Bacillus and other Gram positive bacteria are frequently isolated by this method . The number of colonizer microorganisms on solid media are considerably higher with sediments treated by enzymes, or by enzyme, antibiotic mixtures, than with untreated ones. West Indian Med J, 1982 Dec, 31(4), 198 - 204 Dual intradermal testing of Barbadian children with tuberculin and Battey bacillus antigen; Wells AV et al.; PIP: This article reports on a study comparing 2 different tuberculin tests for effective diagnosis of tuberculosis . 908 Barbadian children, ages 4-8 years were tested simultaneously by intradermal injection of 0.05 g standard tuberculin (PPD--S) and an extract of Mycobacterium intracellulare (Battey bacillus)(PPD--B) . Of the total number of children tested, 21.8% reacted to both antigens . The reaction to PPD--S was significant in 2.6% and to PPD--B in 13.4% . In 5.9% of the children who reacted to 1 or both antigens the reactions were of equal size; the reaction to PPD--S was larger in 17.4% and to PPD--B in 76.7% . It is concluded that infection with atypical mycobacteria is not uncommon in Barbados and that a large number of moderate and small reactions to tuberculin are the result of atypical infection . There is evidence that atypical mycobacterial infection protects against infection with M . tuberculosis and vice versa . No tuberculous infection was found in the children under 4 years of age . In the 4-5, 6-7, 7-8-year age groups, the age specific infection rates were respectively 1.8%, 4.7%, and 4.6% . It is concluded that the policy of administering BDG vaccine to children of 5 years of age should be continued . There is some indication that a number of cases of tuberculosis in Barbados are either not being diagnosed or not being reported . author's modified Int J Lepr Other Mycobact Dis, 1982 Dec, 50(4), 494 - 500 Lack of a sustained protection against murine leprosy in C3H mice vaccinated with extracts of Mycobacterium lepraemurium in admixture with Mycobacterium bovis (BCG); Turcotte R et al.; Inbred C3H mice were vaccinated intradermally with a single dose of live BCG, whole extracts of mechanically disrupted Mycobacterium lepraemurium (MLM), or a mixture of both these agen