Gene, 1985, 40(2-3), 301 - 9 The complete sequence of the Bacillus phage phi 29 right early region; Garvey KJ et al.; We have sequenced the rightmost 2216 bp of the Bacillus phage phi 29 genome . This region encompasses the right early region and completes the sequence of the phi 29 early functions . The sequence of gene 17, an early gene implicated in the replication process, is presented . From these results we predict that gene 17 encodes a 19.1-kDal protein . Further analysis of the sequence revealed five previously undetected potential genes, encoding 12.6-, 12.4-, 15.2-, 6.2- and 4.6-kDal proteins . The biological efficacies of some of these putative genes were demonstrated using an Escherichia coli in vitro transcription-translation system . We also examined the transcriptional and translational signals present on this region of the genome. Mol Gen Genet, 1985, 201(2), 231 - 6 Cloning of the tyrocidine synthetase 1 gene from Bacillus brevis and its expression in Escherichia coli; Marahiel MA et al.; The entire structural gene for tyrocidine synthetase 1 from Bacillus brevis ATCC 8185 has been cloned and expressed in Escherichia coli . Transformed E . coli cells were screened for their ability to produce tyrocidine synthetase 1 by in situ immunoassay using antibodies against gramicidin S synthetase 2 which cross-react with tyrocidine synthetase 1 . The cloned gene is within a 5.2 kb fragment of B . brevis genomic DNA and requires no external promoter for its expression in E . coli . It was also observed that cloning of the 5.2 kb insert in the opposite orientation still resulted in a high level of tyrocidine synthetase 1 expression in transformed E . coli cells . In addition, protein blotting and partial purification of the gene product by gel filtration revealed a major protein of molecular weight about 100,000 with specific D-phenylalanine dependent ATP-32PPi and 2'deoxy ATP-32PPi exchange activities . These unique activities of tyrocidine synthetase 1 were not detected in protein extracts of E . coli strains carrying the vector. Gene, 1985, 36(3), 289 - 300 Characterized full-length and truncated plasmid clones of the crystal protein of Bacillus thuringiensis subsp . kurstaki HD-73 and their toxicity to Manduca sexta; Adang MJ et al.; Bacillus thuringiensis subsp . kurstaki HD-73 produces a crystal protein which is lethal to many lepidopteran larvae . The gene encoding this crystal protein has been isolated from a 75-kb plasmid and engineered into a recombinant Escherichia coli plasmid for analysis . The complete nucleotide sequences of the coding region and 387-bp 5' and 376-bp 3' to the coding region have been determined . The 3537-bp of the coding region specify a protein of Mr 133 330 . The full-length gene and several 3' -truncated derivatives of the gene were examined in both E . coli and in an E . coli minicell-expression system to determine if the carboxy end of the protein is essential for toxicity . The results presented here provide the primary structure of the crystal protein gene and show that the N-terminal 68-kDal peptide is toxic, but at a lower level than the full-length gene product. Gene, 1985, 36(1-2), 169 - 71 Restriction endonuclease mapping of three plasmids from Bacillus thuringiensis var . israelensis; Clark BD et al.; Restriction endonuclease cleavage site maps have been constructed of plasmids pTX14-1, pTX14-2, and pTX14-3 from Bacillus thuringiensis var . israelensis (Bti). J Membr Biol, 1985, 85(3), 199 - 204 Delta endotoxin inhibits Rb+ uptake, lowers cytoplasmic pH and inhibits a K+-ATPase in Manduca sexta CHE cells; English LH et al.; Delta endotoxin, a 68 kilodalton protein isolated from Bacillus thuringiensis spp . Kurstaki, is a potent entomocidal agent that alters a K+ current across midgut tissue of many phytophagous insects . This toxin completely inhibited the vanadate-sensitive 86Rb+ uptake and mimicked the vanadate-induced decrease in cytosolic pH in a cell line (CHE) originating from Manduca sexta embryonic tissue . The toxin also inhibited a K+-sensitive-ATPase in the plasma membranes isolated from these cells . Using the K+-sensitive-ATPase substrate p-nitrophenyl phosphate, delta endotoxin was found to have a Ki of 0.4 microM . These data suggest that the toxin inhibits a K+-ATPase responsible for 86Rb+ uptake in the CHE cells . The relationship between the toxin inhibition of K+-ATPase and toxin-altered K+ current is discussed. Gene, 1985, 34(2-3), 243 - 51 Nucleotide sequence coding for the insecticidal fragment of the Bacillus thuringiensis crystal protein; Shibano Y et al.; The insecticidal crystal protein (ICP) gene, icp, from a 68-kb plasmid derived from Bacillus thuringiensis subsp . sotto was cloned in Escherichia coli . The icp expression in E . coli cells was confirmed by both immunological and insect-toxicity assays of the cell extract . The entire icp gene resides in the 6.6-kb PstI fragment, which codes for a 144-kDal peptide identical to the intact ICP, as determined by its size and reaction with anti-ICP antibody . Deletion analysis further revealed that the 2.8-kb region within the 6.6-kb PstI fragment codes for ICP . Analysis of the nucleotide sequence indicated that a peptide of 934 amino acid residues truncated at the C-terminal end is encoded by this 2.8-kb fragment . A unique feature of this truncated ICP is the abundance of cysteine and lysine residues within its C-terminal region. Mol Gen Genet, 1985, 199(1), 26 - 36 Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces; Bibb MJ et al.; Promoter-probe plasmid vectors were used to isolate putative promoter-containing DNA fragments of three Streptomyces antibiotic resistance genes, the rRNA methylase (tsr) gene of S . azureus, the aminoglycoside phosphotransferase (aph) gene of S . fradiae, and the viomycin phosphotransferase (vph) gene of S . vinaceus . DNA sequence analysis was carried out for all three of the fragments and for the protein-coding regions of the tsr and vph genes . No sequences resembling typical E . coli promoters or Bacillus vegetatively-expressed promoters were identified . Furthermore, none of the three DNA fragments found to be transcriptionally active in Streptomyces could initiate transcription when introduced into E . coli . An extremely biased codon usage pattern that reflects the high G + C composition of Streptomyces DNA was observed for the protein-coding regions of the tsr and vph genes, and of the previously sequenced aph gene . This pattern enabled delineation of the protein-coding region and identification of the coding strand of the genes. Exp Hematol, 1985, 13 Suppl 16, 72 - 9 Relationship between myelosuppression and chemotherapeutic response in small cell bronchogenic carcinoma; Holoye PY; Most cancerocidal agents have myelosuppression as their major toxicity . In some clinical studies it has been possible to show a relationship between the amount of administered drug and the therapeutic efficacy . Within any defined protocol, however, there may be much variability in the severity of myelosuppression . We attempted to determine whether the tumor response might be related to this toxicity . We evaluated a total of 177 patients with small cell bronchogenic carcinoma, treated by five successive regimens of combination chemotherapy, consisting of either cyclophosphamide and vincristine alone or with doxorubicin or doxorubicin plus bacillus Calmette-Guerin (BCG) or doxorubicin plus methotrexate, for a number of prognostic factors (age, sex, extent of disease, performance status, sites and number of metastases, serum LDH and alkaline phosphatase, weight loss, leukopenia, and thrombopenia) . Leukopenia (mean 415 +/- 478/mm3, range 0-2000/mm3) had a weak influence on the incidence of complete remission, which was highest with the least severe nadir (P = 0.027) . Thrombopenia was a nonsignificant factor (P = 0.738) . Both leukopenia and thrombocytopenia had no influence on the overall survival . Because these drug combinations were based on cyclophosphamide, which requires metabolic activation, we evaluated the relationship of myelosuppression and the incidence of response in a second group of patients with small cell bronchogenic carcinoma treated with a VP16, cyclophosphamide, doxorubicin, vincristine sulfate protocol . In this analysis, no relationship could be detected between remission and myelosuppression . Granulocytopenia or thrombocytopenia also-showed no significant influence on the achievement of long-term survival beyond 36 months. Antimicrob Agents Chemother, 1985 Jan, 27(1), 96 - 101 Antibacterial effect of lactoperoxidase and myeloperoxidase against Bacillus cereus; Tenovuo J et al.; An oral periodontopathic bacterium, Bacillus cereus, was inhibited both by lactoperoxidase (LP) and myeloperoxidase (MP) antimicrobial systems . With the LP-SCN--H2O2 system, the growth inhibition was directly proportional to the amount of OSCN- ions present . The OSCN-, which is the principal oxidation product of the LP (or MP)-SCN--H2O2 system at neutral pH, is a normal component of human saliva . The oxidation products of both peroxidase systems inhibited the growth of the bacteria . This inhibition was associated with reduced extracellular release of collagenase activity from the cells . With LP, the antimicrobial efficiency of the oxidizable substrates was SCN- greater than I-, and with MP, the efficiency was I- greater than Cl- greater than SCN-, respectively . LP did not oxidize Cl-. J Bacteriol, 1985 Jan, 161(1), 39 - 46 Delta endotoxin of Bacillus thuringiensis subsp . israelensis; Armstrong JL et al.; From Bacillus thuringiensis subsp . israelensis, a proteinase-resistant protein was purified which exhibited toxicity to larval mosquitoes and cultured mosquito cells, lysed erythrocytes, and was lethal to mice . To extract the protein, a sporulating culture of B . thuringiensis subsp . israelensis was treated with alkali, neutralized, and incubated with trypsin and proteinase K . It was then purified by gel filtration and DEAE column chromatography . Up to 240 micrograms of toxic protein was purified from 1 g (wet weight) of culture pellet . Two closely related forms of toxic protein were obtained: the 25a and 25b proteins . The two forms comigrated near 25,000 daltons in a sodium dodecyl sulfate-polyacrylamide gel, were serologically related, and showed similar partial protease digestion profiles, but were distinguishable by DEAE chromatography and nondenaturing polyacrylamide gel electrophoresis . Protein sequencing data indicated the 25b protein lacked the two amino acids at the amino terminus of the 25a protein . A Western blot enzyme-linked immunosorbent assay of alkali-solubilized proteins that were not treated with proteases suggested the toxic 25a and 25b proteins were proteolytically derived from a larger molecule of about 28,000 daltons . Alkali-solubilized proteins from an acrystalliferous strain of B . thuringiensis subsp . israelensis and from B . thuringiensis subsp . kurstaki failed to cross-react with antibodies to the 25a protein. J Immunol, 1985 Jan, 134(1), 449 - 56 Purification and distribution of a novel macrophage-specific calmodulin-binding glycoprotein; Orlow SJ et al.; The murine macrophage-like cell line J774.16 and peritoneal exudate cells elicited with thioglycollate or starch contain a major calmodulin (CaM)-binding protein (CaMBP) which is absent in trifluoperazine-resistant variants of J774, resident peritoneal macrophages and peritoneal macrophages elicited with concanavalin A, lipopolysaccharide, proteose peptone, or Bacillus Calmette-Guerin . Resident murine peritoneal cells maintained in tissue culture for 3 days begin to accumulate this protein, as do human peripheral blood monocytes after 7 days of culture . A specific competitive displacement radioimmunoassay was developed with the use of a rabbit antiserum raised to the partially purified CaM-binding protein and {125I}CaM covalently cross-linked to the principal CaM-binding protein in the preparation . The radioimmunoassay confirmed the unique cellular distribution of this protein, suggesting that it may be a marker for certain stages of macrophage differentiation . Monoclonal antibodies were prepared, and one of these was used to further purify the protein by immunoaffinity chromatography . A protein of Mr 50,000 to 60,000 was isolated . The protein could be selectively adsorbed to wheat germ agglutinin agarose and subsequently eluted with N-acetyl glucosamine . This property, plus the sensitivity of the protein to endoglycosidase F, led to the conclusion that it is a glycoprotein . The cellular distribution, subcellular localization, and evidence for glycosylation suggest that this protein may be a macrophage-specific receptor with a high affinity for Ca2+-CaM. Allerg Immunol (Leipz), 1985, 31(3), 203 - 6 Age interference of lymphokine production by lung derived lymphocytes; Ganguly R et al.; Young adult and aged guinea pigs, 1 and 3 years old respectively, were sensitized intranasally with Bacillus Calmette Guerin (BCG) . Lung lavage cells were harvested at 2, 3, 6 and 7 weeks following sensitization and tested for migration inhibitory factor (MIF) production in Mackaness chambers with purified protein derivative (PPD) . Oil induced normal pig macrophages were used as indicator cells . Aged guinea pigs, compared to young adult guinea pigs, showed significant delay and reduced levels of MIF production at 2 to 3 weeks . Delayed hypersensitivity skin reaction (DHS) to PPD was also less pronounced at 6 weeks . Senescent immune lung cells were inhibitory to migration of indicator cells in absence of PPD. Cancer Immunol Immunother, 1985, 20(3), 183 - 8 The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives . I . Synthesis of a novel haptenic MDP derivative cross-reactive with Bacillus Calmette Guerin and its application to enhanced induction of tumor immunity; Hamaoka T et al.; A new haptenic compound, a muramyl dipeptide (MDP) derivative (designated as L4-MDP-ONB) cross-reactive with Bacillus Calmette Guerin (BCG) was synthesized . The cross-reactivity of L4-MDP hapten to BCG was demonstrated from the following evidence; (a) lymph node cells from BCG-primed C3H/HeN mice exhibited appreciable L4-MDP-specific proliferative responses to the in vitro stimulation of L4-MDP-modified syngeneic cells (L4-MDP-self); (b) inoculation of L4-MDP-self into footpads of BCG-primed C3H/HeN mice elicited ample delayed type-hypersensitivity (DTH) responses in vivo as measured by footpad swelling; and (c) BCG-primed mice contained L4-MDP-reactive helper T cell activity which functions to augment the generation of effector T cell responses to cell surface antigens . This crossreactivity between L4-MDP hapten and BCG as measured by the helper T cell activity was applied to enhanced induction of tumor-specific immunity . When BCG-primed C3H/HeN mice were immunized with L4-MDP-modified syngeneic X5563 tumor cells, these mice could generate augmented tumor-specific in vivo protective (tumor neutralizing) immunity as well as in vitro cytotoxic T cell responses . These results indicate the effectiveness of L4-MDP hapten in augmenting tumor-specific immunity . The present approach is discussed in the context of potential advantages of this new hapten for its future application to clinical tumor systems. Histopathology, 1985 Jan, 9(1), 109 - 15 Lymph-vessel embolism in a case of Whipple's disease; Adams CW et al.; A case of Whipple's disease is described where the lymphatics in the regional lymph nodes appear to be obstructed by embolized macrophages, containing the characteristic PAS positive bacillary material . It is suggested that the regional lymphangiectasia in Whipple's disease may in part result from such cellular embolism. J Bacteriol, 1985 Jan, 161(1), 47 - 50 Germination-initiated spores of Bacillus brevis Nagano retain their resistance properties; Daher E et al.; Initiated spores and vegetative cells of the gramicidin S-producing Bacillus brevis Nagano were compared with respect to their resistance to various forms of stress (osmotic shock-starvation, exposure to ethanol, sonic oscillation, and heat) . The resistance of initiated spores to all of these stress situations was considerably greater than that of vegetative cells and approached that of dormant spores . The period during which the initiated spores remained resistant to heat was extended by addition of gramicidin S . The antibiotic may therefore be of survival value to the species in nature by slowing down the development of initiated spores in the outgrowth phase of germination, thereby extending the period during which the cells are resistant to environmental stress. Acta Microbiol Pol, 1985, 34(2), 131 - 6 Bacteriophages of Bacillus polymyxa; Starosciak BJ et al.; Virulent bacteriophages of colistin--producing Bacillus polymyxa strains were studied . The phages were found to differ in lytic spectrum and were active only against strains of B . polymyxa . They did not attack other strains of the genus Bacillus . The virulent bacteriophages belong to two morphological groups differing in size . The size of the DNA of the bacteriophages of both groups is similar and ranges from 74.9 X 10(6) to 87.8 X 10(6) daltons . The cells of different B . polymyxa strains were also found to carry various defective phages which could be shown after mitomycin C induction of cell cultures . The antibacterial activity of mitomycin C induced cell lysates was not detected . Strains of B . polymyxa most probably devoid of defective bacteriophages (delysogenized) were isolated. Mikrobiologiia, 1985 Jan-Feb, 54(1), 73 - 82 {Organization and chemical composition of the cell wall of gramicidin S-producing Bacillus brevis}; Simakova IM et al.; The morphology of cells and cell walls was studied in the Bacillus brevis G.-B . R form during its growth and gramicidin S accumulation in it . The membrane apparatus became more complicated and certain other morphological changes were detected in the cells with aging . The cell wall was rather complex even in young cells and consisted of three electron-dense layers where the external and internal layers had an ordered structure . Only the external layer underwent some modifications in the course of growth and these coincided in time with the beginning of intensive gramicidine S biosynthesis . However, the three-layer structure of the cell wall and the ordered organization of the external and internal layers remained unchanged . A preparation of cell walls and preparations of their external and internal layers were isolated from cells synthesizing gramicidine S in the amount of 20 micrograms/ml of the cultural broth . An acid protein having the molecular mass of 100 kD was shown to be the major component of the external layer according to the data of electrophoresis in PAAG with SDS . The middle layer was sensitive to lysozyme, did not have a ordered structure on electron micrographs, and consisted mainly of peptidoglycan. J Chromatogr, 1984 Dec 28, 317, 181 - 92 Application of high-performance liquid chromatographic techniques to the separation of ribosomal proteins of different organisms; Kamp RM et al.; The ribosomal proteins from Escherichia coli, Bacillus stearothermophilus and Methanococcus vannielii were separated by size-exclusion, ion-exchange and reversed-phase high-performance liquid chromatography (HPLC), employing new column materials, different gradient systems, and preparative columns, respectively . The purity of the isolated proteins was analysed by one- and two-dimensional gel electrophoresis and by direct micro-sequencing . The separation of ribosomal proteins could be improved by employing propanol gradients in combination with Vydac reversed-phase columns . From the E . coli ribosome, fifteen S and twenty-three L proteins were isolated in sequencer purity by this method . In addition, ion-exchange HPLC was proven to be useful for isolating ribosomal proteins under native conditions: six S proteins and sixteen L proteins from E . coli could be purified . Some of these proteins were not isolated by the reversed-phase procedures, e.g . proteins L9, L14 and L21. J Biol Chem, 1984 Dec 25, 259(24), 15287 - 93 Probing the structure of 16 S ribosomal RNA from Bacillus brevis; Kop J et al.; A majority (approximately 89%) of the nucleotide sequence of Bacillus brevis 16 S rRNA has been determined by a combination of RNA sequencing methods . Several experimental approaches have been used to probe its structure, including (a) partial RNase digestion of 30 S ribosomal subunits, followed by two-dimensional native/denatured gel electrophoresis, in which base-paired fragments were directly identified; (b) identification of positions susceptible to cleavage by RNase A and RNase T1 in 30 S subunits; (c) sites of attack by cobra venom RNase on naked 16 S rRNA; and (d) nucleotides susceptible to attack by bisulfite in 16 S rRNA . These data are discussed with respect to a secondary structure model for B . brevis 16 S rRNA derived by comparative sequence analysis. Eur J Biochem, 1984 Dec 17, 145(3), 567 - 72 Isolation and the 5'-end nucleotide sequence of Bacillus licheniformis alpha-amylase gene; Sibakov M et al.; We have isolated and determined the 5'-end nucleotide sequence of the alpha-amylase gene from Bacillus licheniformis ATCC 14580 . The alpha-amylase produced by this strain is thermostable and of liquefying type . The gene was originally cloned in a bacteriophage lambda 1059 vector . A subclone containing a 5.3 X 10(3)-base insert in pBR322 was further characterized . The nucleotide sequence coding for the 5' end of the structural gene together with the sequence coding for the upstream control regions was determined . The deduced N-terminal amino acid sequence was identical with the previously published amino acid sequence of B . licheniformis alpha-amylase . There was also very strong homology to the N-terminal sequence of Bacillus amyloliquefaciens alpha-amylase . The Mr of the thermostable alpha-amylase, as determined in vitro in a cell-free transcription/translation system of Escherichia coli, was about 55 000. Eur J Biochem, 1984 Dec 17, 145(3), 645 - 51 Spectroscopic studies of the nature of the iron clusters in the soluble hydrogenase from Desulfovibrio desulfuricans (strain Norway 4); Bell SH et al.; 57Fe-enriched samples of the soluble hydrogenase from Desulfovibrio desulfuricans (Norway) have been investigated in both the native (oxidized) and the dithionite-reduced states using Mossbauer spectroscopy . The data clearly show that the iron in this enzyme is predominantly in the form of iron-sulphur clusters which are closely similar to the {4Fe-4S} clusters found in a large number of ferredoxins, such as that from Bacillus stearothermophilus . There appear to be two {4Fe-4S} clusters . The iron-sulphur clusters in the oxidized protein are virtually diamagnetic, as indicated by Mossbauer, electron spin resonance and magnetic circular dichroic spectroscopy . On reduction by dithionite + methyl viologen, Mossbauer spectroscopy showed that only 50% of the {4Fe-4S} clusters were reduced . Even reduction with hydrogen up to a pressure of 23 GPa did not reduce the iron-sulphur clusters completely . An ESR signal due to a rapidly relaxing species with g = 2.03, 1.89 was observed in the reduced protein, together with a weaker spectrum from a slower-relaxing species at g = 2.34, 2.12. Science, 1984 Dec 14, 226(4680), 1315 - 7 Specific sequence homology and three-dimensional structure of an aminoacyl transfer RNA synthetase; Webster T et al.; Few and limited amino acid sequence homologies have been found among eight bacterial aminoacyl transfer RNA (tRNA) synthetases whose primary structures are known . The entire 939-amino acid primary structure of Escherichia coli isoleucyl-tRNA synthetase is now reported . In a sequence of 11 consecutive amino acids matching a sequence in E . coli methionyl-tRNA synthetase, there are ten identical residues and one conservative change . This is the strongest homology recorded between any two aminoacyl tRNA synthetases . This part of the methionine enzyme's three-dimensional structure has been determined, and it occurs in a mononucleotide binding fold; a close three-dimensional structural homology of this part of the enzyme with Bacillus stearothermophilus tyrosyl-tRNA synthetase has also been reported . The three synthetases probably fold identically in this region. J Biol Chem, 1984 Dec 10, 259(23), 14335 - 6 Ovalbumin is an elastase substrate; Wright HT; Ovalbumin is partially homologous in sequence with the proteinase inhibitors alpha 1-proteinase inhibitor and anti-thrombin III . The region of sequence in ovalbumin which corresponds to the reactive sites of these proteinase inhibitors is susceptible to attack by subtilisin, elastase, thermolysin, bromelain, and Bacillus cereus protease . The esterase activity of elastase is not inhibited by ovalbumin, but ovalbumin is efficiently cleaved by elastase . In contrast with these proteases, trypsin does not cleave ovalbumin. Int J Zoonoses, 1984 Dec, 11(2), 195 - 203 The numbers and varieties of bacteria carried by filth flies in sanitary and unsanitary city area; Adeyemi O et al.; Filth flies caught with nets in various localities of varying sanitary conditions in Ibadan city were predominantly Musca domestica and a few Fannia cannicularis . Seven genera of bacteria, some of which were pathogenic to humans, were isolated from the legs, wings, mouthparts and midguts of the flies . Flies were very abundant in areas where unsanitary conditions prevailed and scarce where sanitary conditions were enforced . The total number of bacteria isolated from flies caught in the market places was higher than those isolated from flies caught in any other locality; low numbers of bacteria were isolated from flies caught in areas where hygienic conditions prevailed . Bacillus spp . were the most numerous of the bacteria isolated . The greatest numbers of bacteria were found on the legs . From the house-flies caught on dairy animals were isolated a high number of Escherichia coli . The public health significance of these findings is discussed. J Biochem Biophys Methods, 1984 Dec, 10(3-4), 163 - 71 Spectrophotometric and radioenzymatic determination of ribose-5-phosphate; Tozzi MG et al.; The present work describes an assay which is highly specific for ribose-5-phosphate . The method is based on the following three-stage enzymatic conversion: (1) ribose 5-phosphate in equilibrium ribose 1-phosphate (phosphopentomutase); (2) ribose 1-phosphate + adenine in equilibrium adenosine + Pi (adenosine phosphorylase); (3) adenosine + H2O----inosine + NH3 (adenosine deaminase) . Ribose 5-phosphate may be determined either directly following the change in absorbance at 265 nm associated with the conversion of adenine to inosine, or radioenzymatically by measuring the radioactivity of inosine formed from {8-14C}adenine, after chromatographic separation of the nucleoside on polyethyleneimine-cellulose . The spectrophotometric assay was used to follow ribose 5-phosphate formation and ribose 1-phosphate consumption catalyzed by phosphopentomutase . Further, the ability of alkaline phosphatase, 5'-nucleotidase and crude extract of Bacillus cereus cells to act on ribose 5-phosphate was tested . The radioenzymatic assay was proved useful in determining the levels of ribose 5-phosphate in rat tissues. J Dairy Sci, 1984 Dec, 67(12), 3081 - 4 Residues in colostrum following antibiotic dry cow therapy; Oliver SP et al.; Cows from five dairy herds were used to determine persistence of antibiotic residues in colostrum and milk following dry cow therapy . Cows were treated in all quarters at drying off with antibiotics approved for use for nonlactating cows . Antibiotics procaine penicillin G plus dihydrostreptomycin, novobiocin, cloxacillin, or cephapirin were compared with no treatment . Composite colostrum samples were collected from each cow at first milking after parturition . Samples were screened for residues by Delvotest P . Colostrum samples positive by Delvotest also were tested by Bacillus stearothermophilus disc assay . Four of 186 colostrum samples from cows treated with antibiotics at drying off were positive for residues by Delvotest . Only one was confirmed positive by disc assay following heat treatment . All colostrum samples from 48 cows not treated were negative . Samples of first marketable milk also were collected . Over 96% of milk samples from cows treated at drying off and 100% of milk samples from cows not treated were negative for residues by Delvotest . If manufacturer's recommendations are followed, antibiotic residues in colostrum and milk following dry cow therapy with products in our study should not be a significant problem. J Appl Bacteriol, 1984 Dec, 57(3), 523 - 30 Influence of NaCl, NaNO2 and oxygen on the germination and growth of Bacillus licheniformis, a spoilage organism of chub-packed luncheon meat; Bell RG et al.; The thermal resistance of Bacillus licheniformis spores was increased from a D70-value of 590 min to one of 900 min by the addition of 4% NaCl to the heating medium {tryptone-yeast extract-glucose (TYG) broth, pH 6.8}, but was decreased to 470 min in TYG broth acidified to pH 4.4 . Sodium nitrite (0.02%) enhanced spore destruction at 80 degrees C but not at 70 degrees C; addition of 4% NaCl eliminated this effect . Less than half the number of spores surviving heat comparable to commercial cooking were heat-damaged to the extent of being unable to grow aerobically in the presence of 4% NaCl . No growth occurred during anaerobic incubation even when the media contained no added NaCl . Oxygen was not required to trigger spore germination, but trace amounts were needed for the successful outgrowth of germinated spores . Spore germination was accelerated and enhanced by the presence of at least 2% NaCl . Therefore under anaerobic conditions NaCl promotes microbiological stability because the germinated spores cannot develop further and become moribund . It is concluded that the plastic casing of luncheon-meat chubs is not sufficiently oxygen-impermeable to allow the product a long shelf-life other than at chill temperatures unless the chubs are stored in an oxygen-free atmosphere. Antimicrob Agents Chemother, 1984 Dec, 26(6), 870 - 5 Single-drug versus combination empirical therapy for gram-negative bacillary infections in febrile cancer patients with and without granulocytopenia; Piccart M et al.; Empirical therapy with cefoperazone was compared with cefoperazone plus amikacin in granulocytopenic and nongranulocytopenic febrile patients . In nonneutropenic patients the overall response rate to cefoperazone was 88%; 10 of 12 gram-negative bacteremic patients were cured . Cefoperazone plus amikacin resulted in an 88% overall response rate and cured 14 of 15 patients with bacteremia . In neutropenic patients the overall response rate was 77% with cefoperazone alone and 73% with cefoperazone plus amikacin; the cure rates for gram-negative bacteremias were 8 of 11 and 6 of 12 patients, respectively . Our findings support the concept of single-drug empirical therapy with cefoperazone in febrile cancer patients, whether granulocytopenic or not, especially when gram-negative bacteremias are predominantly caused by Escherichia coli or Klebsiella species . The issue of Pseudomonas spp . and other more resistant pathogens needs further assessment with a larger number of patients. Antibiotiki, 1984 Dec, 29(12), 922 - 4 {Comparative study of the quantitative determination of kanamycin sulfate in ophthalmic films with a collagen base by microbiological and chemical methods}; Ivanova LA et al.; The results of the comparative study on microbiological and chemical quantitative determination of kanamycin sulfate in the ophthalmic films with the collagen base are presented . The intraocular films prepared with the use of 1 per cent collagen solution contain dexamethasone and kanamycin . The agar diffusion method with Bacillus pumilus NCTC 8241 as the test microbe and the photocolorimetric method based on estimation of the optical density of the colored compound formed after acid hydrolysis of kanamycin with orcinol and ferric chloride were used for the quantitative determination of kanamycin . The results of the quantitative determinations of kanamycin in the films with the two methods did not differ significantly . However, the error of the microbiological method was +/- 3,75 per cent, whereas that of the chemical method was +/- 1.23 per cent or approximately 3 times lower . The time of the analysis decreased from 24 to 1.5-2 h . Moreover, the chemical method is simple and readily reproducible. Gan To Kagaku Ryoho, 1984 Dec, 11(12 Pt 2), 2633 - 9 {Spergualin a novel antitumor antibiotic produced by Bacillus laterosporus}; Takeuchi T; Spergualin, named after the spermidine and guanidine moieties in its structure, was isolated from culture broth Bacillus laterosporus of based on its greater growth-inhibitory activity against chick embryo fibroblasts (CEF) transformed by Rous sarcoma virus compared with that against normal CEF . The structure of spergualin was determined as (-)-(15S)-1-amino-19-guanidino-11, 15-dihydroxy-4, 9, 12-triazanonadecane-10, 13-dione . Spergualin is water-soluble, basic and a white powder . It shows cytotoxicity in vitro against mouse leukemias L1210 and L5178Y . It significantly prolongs the life span of mice transplanted with either of the above leukemic cells . It induces a strong and specific immunological effect on L1210 which has been maintained in our laboratory; L1210-transplanted CDF1 mice, once cured by spergualin, reject a second challenge of the leukemic cells . Spergualin hibits in growth in vitro of both Gram (+) and (-) bacteria at concentrations ranging from 6.25 to 100 micrograms/ml. J Lab Clin Med, 1984 Dec, 104(6), 908 - 20 Effects of iron loading and bacillus Calmette-Guerin on a glycoprotein recognition system on rat hepatic sinusoidal cells; Parise ER et al.; Experiments were performed to determine the effects of agents that modify Kupffer cells on the mannose-N-acetylglucosamine-glycoprotein receptor on hepatic sinusoidal cells . Cells were prepared by collagenase liver perfusion, centrifugation on Percoll gradients, and centrifugal elutriation . The uptake of 125I-labeled agalacto-orosomucoid (125I-AGOR), an N-acetylglucosamine-terminated glycoprotein, was greatest (53% of total uptake) by elutriator fractions containing equal proportions of endothelial and Kupffer cells ("mixed cell" fraction) . Uptake was specific and time and concentration dependent . The apparent Km (0.4 mumol/L) and the patterns of inhibition by monosaccharides were similar in all the elutriator fractions, suggesting that only one class of receptor was present . The highest apparent maximal velocity (18 pmol/hr/5 X 10(6) cells) was found in the mixed cell fraction, indicating this fraction contained the highest proportion of receptor-bearing cells . Latex (0.8 micron) and bacillus Calmette-Guerin pretreatments did not influence the hepatic uptake of the glycoprotein in vivo . Iron sorbitol significantly reduced hepatic glycoprotein uptake and caused a twofold increase in the proportion of the ligand remaining in the circulation . Uptake of 125I-agalacto-orosomucoid by cells from latex-treated rats was similar to controls, but uptake by bacillus Calmette-Guerin-treated rat cells was only 25% of control uptake . This was related to a marked increase in sinusoidal cell number caused by bacillus Calmette-Guerin . In contrast, iron sorbitol caused a selective suppression of 125I-agalacto-orosomucoid uptake (10% of control uptake) by cells in the mixed cell fraction . This study showed that maximal uptake of 125I-agalacto-orosomucoid was by elutriator fractions containing equal proportions of endothelial and Kupffer cells and that iron sorbitol suppressed ligand uptake by these cells, possibly by influencing the mannose-N-acetylglucosamine receptor on Kupffer cells. Fed Proc, 1984 Dec, 43(15), 2981 - 3 Crystallization of substrate and product analog complexes of tryptophanyl-tRNA synthetase; Carter CW Jr et al.; We have prepared crystals of tryptophanyl-tRNA synthetase from Bacillus stearothermophilus complexed to tryptophan (type II*), and to tryptophanyl-3'(2')-ATP (type IV) . The latter compound is a product analog, enzymatically synthesized by acyl transfer of tryptophan from the tryptophanyl-5'-AMP intermediate to a second molecule of ATP . It resembles the 3'-terminal fragment, tryptophanyl-3'(2')-adenosine, of Trp-tRNATrp . Both crystal forms diffract to high resolution . Although both forms are grown from 2 M K2HPO4, they are dramatically different in the shape of the unit cell and in space group symmetry . Type II* crystals are monoclinic (space group P21) . However, low-resolution reflections obey the symmetry of space group P321, which indicates both the existence and the location of noncrystallographic symmetry in the monoclinic unit cell . Type IV crystals belong to space group P41212 (or its enantiomorph) and the unit cell is elongated along the fourfold screw axis . Analysis of molecular packing suggests that intermolecular contacts in the two crystal types are very different . Thus, the two structures may exhibit conformational differences related to catalysis by this enzyme . Solution of type II* and type IV crystal structures may provide representations resembling a Michaelis complex and an acyl transfer product complex. J Appl Bacteriol, 1984 Dec, 57(3), 447 - 54 An improved enzyme-linked immunoassay for the detection and quantification of the entomocidal parasporal crystal proteins of Bacillus thuringiensis subsp . kurstaki and israelensis; Wie SI et al.; An improved and simplified enzyme-linked immunosorbent assay (ELISA) was developed for the detection and quantification of parasporal crystalline toxins from Bacillus thuringiensis subsp . kurstaki . The improved procedure involved pretreatment of the polystyrene cuvettes with glutaraldehyde before antibody coating . A direct comparison of treated and untreated cuvettes is provided . ELISAs were then used for the analysis of the entomocidal crystalline proteins in commercial and experimental formulations of B . thuringiensis subspp . kurstaki and israelensis. Hoppe Seylers Z Physiol Chem, 1984 Dec, 365(12), 1445 - 9 An ultracentrifuge study on the self-association of glucose dehydrogenase from Bacillus megaterium; Schubert D et al.; The self-association of glucose dehydrogenase (beta-D-glucose:NAD(P) 1-oxidoreductase, EC 1.1.1.47) from Bacillus megaterium was studied by analytical ultracentrifugation . The pH and composition of the buffer used were such that, owing to a reversible partial dissociation of the tetrameric enzyme, enzyme activity was reduced . It was found that under these conditions the protein exists in a monomer/dimer/tetramer association equilibrium. Antimicrob Agents Chemother, 1984 Dec, 26(6), 939 - 40 Mycocerein, a novel antifungal peptide antibiotic produced by Bacillus cereus; Wakayama S et al.; A peptide was obtained from culture filtrates of a bacterium which was newly isolated and tentatively named Bacillus cereus SW . The peptide was composed of Asx, Ser, Glx, Leu, Tyr, Pro, and an unknown amino acid in a ratio of 2:1:1:1:1:1:1, but, unless hydrolyzed with HCI, it was ninhydrin reaction negative . The peptide effectively inhibited the growth of all fungi and yeasts so far examined, whereas it inhibited none of the bacteria tested. Inflammation, 1984 Dec, 8(4), 393 - 406 Glucocorticoid modulation of lymphokine-induced giant cell formation; Galindo B; Lymph node lymphocytes from rabbits sensitized with bacillus Calmet Guerin (BCG) secreted into the culture media both macrophage fusion factor (MFF) and migration inhibition factor (MIF) after 24 h of incubation with heat-killed BCG . Cell-free supernatant fluids obtained from these cultures induced simultaneously giant cell formation and migration inhibition of homologous normal alveolar macrophages . The glucocorticoids cortisol (10(-7) M) and dexamethasone (10(-8) M) (DX) consistently inhibited giant cell formation elicited by MFF (P = 0.003) without affecting macrophage viability . By contrast, the same glucocorticoids, in concentrations ranging from 10(-8) to 10(-10) M, induced a considerable increment of giant cell development in macrophage populations exhibiting a low response to MFF . Neither cortisol (10(-4) M) nor DX (10(-4) M) affected the migration inhibition of alveolar macrophages induced by MIF . Present results suggest that the granulomatous response in the rabbit, as reflected by the macrophage fusion assay, may be regulated by glucocorticoids. Clin Exp Immunol, 1984 Dec, 58(3), 531 - 8 Inhibition of interleukin-2 production by adherent cell factors from lepromatous leprosy patients; Nath I et al.; Twenty-four hour supernatants (MoF) were obtained from monocyte rich 2 h adherent cells of 19 leprosy patients and four healthy contacts . MoF from borderline and lepromatous patients produced 52-61% inhibition of human interleukin-2 (IL-2) production by a PHA conditioned T cell line (Jurkat) . Non-adherent cell supernatants and MoF from tuberculoid and healthy individuals had little effect on IL-2 production . The suppression effected by MoF was in the first 12 h of initiation of PHA stimulated Jurkat cell cultures . Suppressive MoF did not interfere with (1) IL-2 release, (2) IL-2 utilization by Con A-induced T cell blasts or (3) constitutive proliferation of Jurkat cells . Such MoF were released spontaneously from adherent cells of bacilliferous leprosy patients but required in vitro antigen triggering in long term treated lepromatous patients . It is possible that the unresponsiveness associated with lepromatous leprosy is related to the inhibition of IL-2 production by suppressive factors, thereby, preventing the further expansion of antigen reactive T cells. J Exp Med, 1984 Dec 1, 160(6), 1686 - 701 Production of target-specific effector cells using hetero-cross-linked aggregates containing anti-target cell and anti-Fc gamma receptor antibodies; Karpovsky B et al.; Rabbit anti-2,4-dintrophenyl (DNP) antibodies or their F(ab')2 fragments were chemically cross-linked to the anti-mouse Fc gamma R monoclonal antibody 2.4G2 or to its Fab fragment . P388D1 cells were incubated with heteroaggregates between 2.4G2 and anti-DNP (anti-Fc gamma R X anti-DNP) and washed . The resulting cells lysed 2,4,6-trinitrophenyl chicken erythrocytes (TNP CRBC) in a hapten-specific manner . The lysis was inhibited by free hapten but was resistant to inhibition by immune complexes . Other cells coated with antibody heteroaggregates also mediated lysis of TNP-modified target cells . For example, mouse resident peritoneal exudate cells (PEC) lysed TNP CRBC and bacillus Calmette-Guerin-activated PEC lysed both TNP CRBC and TNP tumor targets . Human neutrophils, when incubated with heteroaggregates containing the anti-human neutrophil Fc gamma R antibody 3G8 and anti-DNP also lysed TNP CRBC and TNP-modified tumor cells . To test whether linkage to Fc gamma R was required for lysis, F(ab')2 fragments from the anti-KdDd monoclonal antibody 34-1-2 were cross-linked to anti-DNP F(ab')2 fragments . P388D1 cells (which express Kd and Dd) were then incubated with these heteroaggregates and washed, and their abilities to form conjugates and lyse TNP CRBC were compared with those of P388D1 cells treated with anti-Fc gamma R X anti-DNP . In both cases, P388D1 cells formed conjugates . However, only the cells treated with anti-Fc gamma R X anti-DNP mediated lysis to a significant extent . We conclude that heteroaggregates containing anti-Fc gamma R and anti-target cell antibodies can be used to create potent effector cells against red cell and tumor targets and that bridging of effectors with target cells directly to Fc gamma R on effector cells is required for lysis. J Bacteriol, 1984 Dec, 160(3), 854 - 9 Molecular cloning of structural and immunity genes for megacins A-216 and A-19213 in Bacillus megaterium; Von Tersch MA et al.; A host-vector system was developed for molecular cloning in Bacillus megaterium and used to clone the structural and immunity genes for megacins A-216 and A-19213 . Recombinant clones that expressed immunity only or both immunity to and production of each megacin were obtained . Restriction mapping of native megacinogenic plasmids and recombinant clones was used to construct physical and genetic maps of megacinogenic plasmids pBM309 and pBM113 . Limited sequence homology between pBM309 and pBM113 was detected by Southern blot hybridization and was mapped to, at most, a 6.4-kilobase-pair region of pBM309 and a 6.1-kilobase-pair region of pBM113. Cancer, 1984 Dec 1, 54(11 Suppl), 2751 - 65 Infusion-embolization; Wallace S et al.; Transcatheter intra-arterial therapy for the cancer patient encompasses infusion of chemotherapy and embolization . Intra-arterial infusion of chemotherapeutic agents has been resurrected because of the availability of new drugs, combinations of drugs, and the capability of percutaneous selective catheter placement . Intra-arterial infusion has been effective in patients with carcinomas of the liver, bladder, prostate, uterus, ovary, and lung and in bone and soft tissue sarcomas, melanomas, and tumors of the brain . Embolization of the arterial supply, creating ischemia of the neoplasm, has been employed in the therapeutic management of patients with primary and secondary neoplasms of the liver, kidney, and bone . The median survival of 100 patients with neoplasms of the liver from the time of hepatic artery embolization was 11.5 months . In 100 patients with pulmonary metastases from carcinoma of the kidney, 28 experienced a response to renal artery embolization, a therapeutic delay of 4 to 7 days, nephrectomy, and Depo-Provera (medroxyprogesterone) . Seven of 12 patients with giant cell tumor of the pelvis and lumbar spine responded to arterial embolization after all other therapy failed . Chemoembolization, the combination of arterial infusion of chemotherapy and embolization, can be accomplished by the use of microencapsulated agents, liposomes, and particulate emboli with drugs . This approach integrates the advantages of infusion and occlusion, and has considerable potential . Intra-arterial immunotherapy has been initiated with bacillus Calmette-Guerin (BCG) administration into renal neoplasms in patients with metastatic disease. J Biol Chem, 1984 Nov 10, 259(21), 13590 - 4 Identification and characterization of some bacterial membrane sulfhydryl groups which are targets of bacteriostatic and antibiotic action; Morris SL et al.; Covalent modification of sulfhydryl groups which become sensitive toward sulfhydryl agents during germination of Bacillus cereus spores exerts a profound bacteriostatic effect, resulting in outgrowth inhibition . The modified spore components are membrane species of 13,000, 28,000, and 29,000 daltons . Detergent disruption of the membrane inactivated the sulfhydryl groups . A highly sigmoid inhibition curve (n = 11.8) with diamide suggested the participation of closely neighboring sulfhydryl groups . Substate and substrate analogs of the lactose and dicarboxylic acid permeases protected the sulfhydryl groups against modification . Nisin, a 34-residue peptide antibiotic, inhibited spore outgrowth and sulfhydryl modification at a concentration of about 0.1 microM . Since these sulfhydryl groups have been implicated as involved with the bacteriostatic action of nitrite, substances directed toward them may be a useful new class of bacteriostatic agents and antibiotics. Antimicrob Agents Chemother, 1984 Nov, 26(5), 730 - 3 Penetration of aztreonam into cerebrospinal fluid of patients with and without inflamed meninges; Duma RJ et al.; Aztreonam was administered as a single, 2-g intravenous dose to 25 patients with noninflamed meninges and to 9 patients with inflamed meninges . It was well tolerated and was detected in the cerebrospinal fluid at the initial sampling period at 1 h after the end of infusion . Aztreonam levels in the cerebrospinal fluid of patients with inflamed meninges were four times higher than those recorded for the same time period in patients with noninflamed meninges . Aztreonam concentrations in cerebrospinal fluid in the presence of normal and inflamed meninges exceeded the inhibitory and bactericidal concentrations for most gram-negative bacteria . Thus, a multiple-dose treatment regimen with 2-g intravenous doses every 6 h appears to be appropriate for clinical trials of aztreonam for the treatment of gram-negative bacillary meningitis which is caused by susceptible organisms. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1984 Nov, 17(4), 233 - 40 Metabolites of Bacillus sp . isolated from flax-suppressive soil; Yang SS et al.; A culture of Bacillus sp . was isolated from the suppressive flax soil . It could inhibit the growth of many plant pathogens that were found in suppressive flax soil, and an acidic ethyl acetate extract of culture broth of the isolated Bacillus showed significant inhibition of the growth of some plant pathogens . The antimicrobial activity of culture extracts (15 times concentrated) was higher than that of 400 micrograms/ml actidione or 200 micrograms/ml mycostatin (nystatin) . The antimicrobial substances were relatively stable . After heating at 100 degrees C for 10 min . and 30 min., the activities were still 100% and 72%, respectively . Properties of the compounds were investigated by UV, TLC, GC and GCMS . The component with the retention time of 13.4 min in a gas chromatography was biologically active and the parent peak was at m/z 142. Ann Clin Lab Sci, 1984 Nov-Dec, 14(6), 464 - 6 The effect of BCG-vaccine upon experimental visceral leishmaniasis in hamsters; Jarecki-Black JC et al.; Stimulation with Bacillus Calmette-Guerin (BCG) vaccine has been reported to enhance resistance of mice against Leishmania donovani infection . Such infection is usually lethal in hamsters, thus providing a more stringent animal model to assess the effect of BCG upon visceral leishmaniasis . Animals receive two IP injections (2-8 X 10(7) BCG) pre or post IC challenge with 4 X 10(6) amastigotes . Controls received BCG alone (with no infection) or were untreated (NT) . Pretreated animals exhibited significantly fewer (P less than 0.05) hepatic or splenic amastigotes than NT animals at days 7, 14, and 28 post challenge, but most BCG treated hamsters died earlier than NT . Post treated hamsters showed no significant reduction in parasite burdens, or in median time to death as compared to NT group . Hamsters which received BCG but were not infected appeared healthy during the study . The reason for increased susceptibility of BCG-treated hamsters to disease is not clear, but observed pathologic complications of L . donovani infected hamsters appear to be exacerbated by BCG stimulation. J Bacteriol, 1984 Nov, 160(2), 591 - 9 Purification and properties of glutamate synthase from Bacillus licheniformis; Schreier HJ et al.; Glutamate synthase {L-glutamate:NADP+ oxidoreductase (transaminating); EC 1.4.1.13}(GltS) was purified to homogeneity from Bacillus licheniformis A5 . The native enzyme had a molecular weight of approximately 220,000 and was composed of two nonidentical subunits (molecular weights, approximately 158,000 and approximately 54,000) . The enzyme was found to contain 8.1 +/- 1 iron atoms and 8.1 +/- 1 acid-labile sulfur atoms per 220,000-dalton dimer . Two flavin moieties were found per 220,000-dalton dimer, with a ratio of flavin adenine dinucleotide to flavin mononucleotide of 1.2 . The UV-visible spectrum of the enzyme exhibited maxima at 263,380 and 450 nm . The GltS from B . licheniformis had a requirement for NADPH, alpha-ketoglutarate, and glutamine . Classical hyperbolic kinetics were seen for NADPH affinity, which resulted in an apparent Km value of 13 microM . Nonhyperbolic kinetics were obtained for alpha-ketoglutarate and glutamine affinities, and the reciprocal plots obtained for these substrates were biphasic . The apparent Km values obtained for glutamine were 8 and 100 microM, and the apparent Km values obtained for alpha-ketoglutarate were 6 and 50 microM . GltS activity was found to be relatively insensitive to inhibition by amino acids, keto acids, or various nucleotides . L-Methionine-DL-sulfoximine, L-methionine sulfone, and DL-methionine sulfoxide were found to be potent inhibitors of GltS activity, yielding I0.5 values of 150, 11, and 250 microM, respectively . GltSs were purified from cells grown in the presence of ammonia and nitrate as sole nitrogen sources and were compared . Both yielded identical final specific activities and identical physical (UV-visible spectra, flavin, and iron-sulfur composition) and kinetic characteristics. Gut, 1984 Nov, 25(11), 1203 - 10 Surface morphology of the gastroduodenal mucosa in duodenal ulceration; Steer HW; Endoscopic biopsies from the duodenal cap and prepyloric areas of 25 patients have been examined with the scanning electron microscope . Eleven patients had duodenal ulceration . Bacteria are related only to the surface of gastric type epithelial cells whether these cells are located at areas of gastric metaplasia in the duodenal bulb or in the pre-pyloric region of the stomach . The bacteria are not associated with the surface of intestinal type epithelial cells . The bacteria are absent from the biopsies of those patients with a normal stomach and duodenum . Of those patients with duodenal ulceration, 73% have bacteria related to the epithelial surface . The bacteria are of two morphological types - a kidney shaped bacillus and an S-shaped bacillus. Radiology, 1984 Nov, 153(2), 311 - 6 Thoracic tuberculosis, mycobacteriosis, MERosis, and BCGosis in a cancer treatment center; Hill CA; Radiographs and clinical records of 100 patients with tuberculosis or mycobacteriosis and six patients infected with Bacillus Calmette-Guerin (BCG) or methanol extraction residue of BCG (MER) were reviewed . Radiographic presentations were categorized . Tuberculosis or mycobacteriosis simulated pulmonary neoplasm in 64 patients . Tuberculosis or mycobacteriosis complicated pulmonary neoplasm, hematological neoplasm, or acquired immune deficiency syndrome in 27 . Coincident tuberculosis or mycobacteriosis and bronchogenic carcinoma were encountered in seven . Bronchogenic carcinoma developed in an area of chronic stable tuberculosis in three . Miliary granulomatous lesions developed in four following MER immunotherapy and in two following BCG immunotherapy . Differentiating tuberculosis or mycobacteriosis from bronchogenic carcinoma, metastasis, or pneumonia was difficult and biopsy was often necessary. J Clin Microbiol, 1984 Nov, 20(5), 837 - 42 Oxygen metabolism in phagocytes of leprotic patients: enhanced endogenous superoxide dismutase activity and hydroxyl radical generation by clofazimine; Niwa Y et al.; We examined the generation of active oxygens (O2-, H2O2, and OH X ) and the superoxide dismutase (SOD) activity of polymorphonuclear leukocytes (PMNs) and monocytes from 14 leprotic patients manifesting a bacillary index above 2.2 . Patients with disease of more than 4 years in duration showed significantly enhanced SOD activity and a decrease in O2- and OH X production . The antileprotic agent, clofazimine, significantly increased the generation of OH X in a dose-dependent manner, with a subsequent decrease in H2O2, but had no effect on the SOD activity of the PMNs and monocytes . In medium containing FeSO4 or Fe2+-EDTA, the drug elevated OH X production markedly further . Phagocytic SOD in PMNs and monocytes of leprotic patients was both host and bacillus derived, because the presence of cyanide, to which human-derived cuprozinc SOD is susceptible, did not completely abrogate SOD activity . The difficulty in treating leprosy may be partly ascribable to decreased phagocytic OH X generation, which in leprosy patients is apparently due to the uptake of Hansen bacillus-derived SOD . Clofazimine may be effective in leprosy by chelating Fe2+, with the resultant potentiation of the catalyzing activity of Fe2+ in the Haber-Weiss reaction increasing OH X formation from H2O2. Mikrobiologiia, 1984 Nov-Dec, 53(6), 970 - 5 {An oligosporogenic mutant of Bacillus brevis}; Zarubina AP et al.; The treatment of Bacillus brevis var . G-B by chemical mutagens yielded a mutant similar in the major characteristics to the parent strain of the P+-variant, but producing less spores and synthesizing more gramicidin S . The mutant can be stored in the freeze-dried state. J Biol Chem, 1984 Oct 25, 259(20), 12602 - 8 Ammonia/potassium exchange in methanogenic bacteria; Sprott GD et al.; Methanospirillum hungatei exposed to ammonia in a K+-free buffer lost up to 98% of the cytoplasmic K+ through an ammonia/K+ exchange reaction . The exchange was immediate, and occurred in cells poisoned by air or by other metabolic inhibitors . Additions of NH4OH or various NH+4 salts (or methylamine) were most effective in causing K+ depletion in media of alkaline pH, suggesting that NH3 was the chemical species crossing the membrane . In alkaline media, the exchange reaction resulted in a dissipation of the transmembrane pH gradient (inside acidic), but had only small effects on the membrane potential until concentrations of ammonia were used above those required to abolish the K+ gradient . Through the use of NH4Cl to vary the cytoplasmic pH at a constant acidic external pH, and NH4OH to abolish the transmembrane pH gradient at various alkaline external pH values, we conclude that methanogenesis is sensitive to both the pH of the cytoplasm and the medium . Methanogenesis in Msp . hungatei and Methanosarcina barkeri was inhibited dramatically at external pH values more acidic than 6.5 or more alkaline than 7.5 . Dramatic K+ depletion in response to ammonia additions at pH 8.0 occurred with Ms . barkeri, another strain of Msp . hungatei, Escherichia coli, and Bacillus polymyxa . In several other methanogens, ammonia/potassium exchange was hardly detected. Pharm Weekbl Sci, 1984 Oct 19, 6(5), 209 - 15 Microbiological aspects of heat sterilization of medicines . II . A method for the determination of the effectiveness of a sterilization process using the bioburden and the bioburdens heat resistance; Boom FA et al.; In order to verify whether the sterilization process of 60 min at 100 degrees C for invert sugar 20% is sufficiently effective to attain the generally accepted probability of survival of maximum 1 X 10(-6), we determined the bioburden and the bioburdens heat resistance for this product . We examined 98 bottles by the membrane filtration method and found 84 bottles with 0 colony forming units (CFU's) and 14 bottles with 1-9 CFU's . Because none of the isolated CFU's was heat resistant (Bacillus species), we isolated heat resistant CFU's from the environment and determined the heat resistance in invert sugar, water and NaCl solution 0.9% of four different Bacillus species . The results in invert sugar for the most heat resistant Bacillus species were a D-value of 0.92 min at 100 degrees C . For the determination of the D-value the end-point method is the most practical one, and the D-value calculation with the most probable number method is sufficiently accurate . Because of unavoidable inaccuracies in the experimentally determined D-value, safety margins of 100% have to be taken into account in the sterilization process calculations in which these D-values are used . Hence, in our case, we have to use a D-value of 2 X 0.92 min in the sterilization process calculation for invert sugar 20% . The maximum bioburden in the examined 98 test bottles was 9 CFU's . The maximum heat resistant bioburden which must be used in sterilization process calculations may be safely fixed at 10% of the total bioburden, therefore we have to use 0.9 micro-organisms in our calculation.(ABSTRACT TRUNCATED AT 250 WORDS) Biochim Biophys Acta, 1984 Oct 16, 801(3), 372 - 80 Acylureas: a new class of barbiturate-like bacterial cytochrome P-450 inducers; Ruettinger RT et al.; The soluble, cytochrome P-450-dependent fatty acid monooxygenase of Bacillus megaterium ATCC 14581 is induced by phenobarbital and at least twelve other barbiturates (Kim, B.-H . and Fulco, A.J . (1983) Biochem . Biophys . Res . Commun . 116, 843-850) . We have since found that the inducer potency of phenobarbital and of six other of these barbiturates was enhanced by adding them to the growth medium prior to sterilization by autoclaving . A similar 'activation' was effected simply by autoclaving these barbiturates in distilled water at pH 8.0 . When the hydrolytic products resulting from such treatment of phenobarbital were identified and screened for inducer activity, the major product, 2-phenylbutyrylurea, was found to be 3-5-times more potent than phenobarbital itself . The racemic mixture, (+/-)-2-phenylbutyrylurea was somewhat more active as an inducer than was either of the enantiomers {+/-) or (-} tested singly . Of the other hydrolytic products of phenobarbital, only 2-phenylbutyramide had significant inducer activity (about the same as phenobarbital) . Among other ureides tested, two monosubstituted acetylureas (phenylacetylurea and dodecanoylurea) were inactive as inducers, but six of seven disubstituted acetylureas were better inducers than 2-phenylbutyrylurea. Cell Immunol, 1984 Oct 15, 88(2), 285 - 96 Acquired immunity to heavy infection with Mycobacterium bovis bacillus Calmette-Guérin and its relationship to the development of nonspecific unresponsiveness in vitro; Orme IM et al.; Mice heavily infected with Mycobacterium bovis bacillus Calmette-Guerin (BCG) rapidly generated an acquired cellular immune response to this infection, as characterized primarily by the emergence of a splenic T-cell population capable of passively transferring substantial levels of adoptive protection against a challenge infection with M . tuberculosis . The emergence of this protective T-cell population was temporally associated with considerable levels of DNA synthesis in vivo in both the spleen and liver, and with the development of an acquired capacity within the animal to express very high levels of nonspecific resistance to secondary intracellular bacterial infection . Concomitant with the emergence of this acquired response, splenic T cells from infected animals became severely unresponsive to blastogenic in vitro stimulation with the mitogen phytohemagglutinin, and possessed the capacity to suppress the responsiveness of normal T cells in cocultures . Both the unresponsiveness of T cells from infected mice and their immunosuppressive activity in vitro could be essentially ablated by supplementation of the tissue culture medium with a supernatant containing very high titers of the T-cell growth factor interleukin 2 (IL-2) . Furthermore, T cells harvested from these animals at the peak of in vitro unresponsiveness exhibited a substantial capacity to absorb or consume IL-2 from IL-2-containing supernatants . It is hypothesized, on the basis of these findings, that mice heavily infected with BCG acquire an IL-2-dependent T-cell population within the spleen in response to this infection, and that the observed in vitro blastogenic unresponsiveness of spleen cells which contain this population may be an artefactual effect arising from the reduction or consumption of available IL-2 within the sustaining culture medium . The relevance of these findings is discussed with particular regard to clinical situations, such as lepromatous leprosy, in which restorative strategies involving the in vivo use of IL-2 are presently being postulated. Food Addit Contam, 1984 Oct-Dec, 1(4), 349 - 58 Excretion of penicillins in bovine milk following intramuscular administration; Moretain JP et al.; The kinetics of elimination into milk of sodium penicillin G, procaine penicillin G, benzathine penicillin G, ampicillin and amoxycillin residues have been determined after intramuscular administration of eleven drugs chosen among those commercially available in France . These investigations will be used as a basis to estimate and harmonize the withdrawal times demanded for veterinary drugs . The quantitative analysis of residues was carried out by a cylinder plate microbiological method with Bacillus stearothermophilus as test organism . The threshold of detection is 0.001 unit (or micrograms)/ml of milk . The mean durations of elimination are four milkings for the association sodium penicillin G and procaine penicillin G, from seven to eight milkings for procaine penicillin G alone, from 19 to 33 milkings for benzathine penicillin G, and from three to five milkings for ampicillin and amoxycillin. Cell, 1984 Oct, 38(3), 835 - 40 The use of double mutants to detect structural changes in the active site of the tyrosyl-tRNA synthetase (Bacillus stearothermophilus); Carter PJ et al.; In a previous study, a mutant of tyrosyl-tRNA synthetase in which a threonine residue (Thr51) was converted to proline dramatically improved the affinity of the enzyme for its ATP substrate . How does Pro51 improve the enzyme's affinity for ATP? A priori, Pro51 might interact directly with the ATP, or it might distort the polypeptide backbone and thereby force new or improved contacts elsewhere from the enzyme to ATP . By making mutants of the Pro51 enzyme at two residues that make hydrogen bonds to the ATP substrate, we show that Pro51 greatly improves the strength of one of these contacts . Thus the propagation of a structural change in an enzyme induced by mutation may be detected by the introduction of further mutations. J Urol, 1984 Oct, 132(4), 675 - 7 Bacillus Calmette-Guerin immunotherapy of infiltrating bladder cancer; Netto NR Jr et al.; Oral administration of bacillus Calmette-Guerin was used to treat 10 patients with muscle invasive transitional cell carcinoma of the bladder . The treatment induced tumor regression in 7 patients (70 per cent), including 1 who died without evidence of recurrent tumor . Skin test reactivity was correlated with response to treatment . Only 1 patient had marked skin test reactivity at the time of tumor recurrence . Our data demonstrated that patients with invasive bladder cancer may derive benefit from immunotherapy. Clin Pediatr (Phila), 1984 Oct, 23(10), 586 - 9 Presumed BCG infection in a boy with chronic granulomatous disease . A report of a case and a review of the literature; Kobayashi Y et al.; A male child with chronic granulomatous disease (CGD) developed protracted axillary lymphadenopathy with liquefaction following a bacille Calmettle-Guerin (BCG) immunization . Except for phagocytic dysfunctions characteristic of the underlying disease, immunological examinations were normal . The literature dealing with CGD cases with disseminated BCG infection was reviewed . It is concluded that the possibility of CGD should be considered in those who developed such adverse reactions to BCG immunization. Gene, 1984 Oct, 30(1-3), 167 - 72 Bacillus megaterium spore protein C-3: nucleotide sequence of its gene and the amino acid sequence at its spore protease cleavage site; Fliss ER et al.; The nucleotide sequence of the Bacillus megaterium gene coding for spore-specific protein C-3 has been determined . The gene codes for 65 amino acids and the coding sequence is preceded by an efficient ribosome-binding site . The predicted protein C-3 sequence agrees with both the amino acid composition and the amino terminal sequence of protein C-3, and shows homology (approx . 65% of all residues are identical) with the sequences of the analogous proteins A and C of B . megaterium . Protein C-3 is cleaved by the sequence-specific B . megaterium spore protease, and the amino acid sequence at the new amino-terminus generated is identical to that predicted from the gene sequence, and homologous to the spore protease cleavage sites in the A and C proteins . The protein C-3 gene also shares a number of features with the previously sequenced protein C gene in both upstream and downstream flanking sequence. Arch Biochem Biophys, 1984 Oct, 234(1), 61 - 72 The pH dependence and group modification of beta-D-xylosidase from Bacillus pumilus: evidence for sulfhydryl and histidyl groups; Kersters-Hilderson H et al.; The pH dependence of the kinetic parameters of beta-D-xylosidase (EC . 3.2.1.37) from Bacillus pumilus reveals that an acidic functional group with pK 8.0 is involved in the catalysis . The fast inactivation of the dimeric enzyme by near equivalent amounts of methylmethanethiolsulfonate indicates that one thiol group per monomer is essential for catalysis, consistent with previously reported results . From the reactivity of the thiol groups with respect to 5,5'-dithiobis(2-nitrobenzoic acid), the absence of subunit cooperativity was indicated . The present study also reports on the inactivation of the enzyme by diethylpyrocarbonate, and provides evidence of the importance of a histidine residue . A mechanism of catalysis is presented, in which the thiol group interacts with the substrate via partial proton transfer . The mode of participation of the histidine group is difficult to specify, but may be associated with the maintenance of the active conformation of the enzyme. J Bacteriol, 1984 Oct, 160(1), 473 - 7 Respiratory system of vegetative and sporulating Bacillus cereus; Escamilla JE et al.; The composition and organization of the Bacillus cereus respiratory system were studied . The abolition of NADH-dependent respiration in vegetative and sporulating cell membranes by near-UV light (360 nm) indicated that electrons reduce oxygen only through a quinone-cytochrome pathway . Difference spectroscopy demonstrated the presence of cytochromes b555, c548, aa3, b562, and a2 . This composition and studies with respiratory inhibitors suggested that cytochromes are organized in at least two branches, one being highly sensitive to cyanide. Acta Leprol, 1984 Oct-Dec, 2(2-4), 394 - 402 IgM antibodies against phenolic glycolipid I from Mycobacterium leprae in leprosy sera: relationship to bacterial index and erythema nodosum leprosum; Schwerer B et al.; Serum IgM antibodies against Mycobacterium leprae-derived phenolic glycolipid I (PG) were determined in 121 leprosy patients, in contacts and controls by an enzyme-linked immunosorbent assay technique . Anti-PG IgM levels correlated with disease classification, increasing from the tuberculoid towards the lepromatous pole of the disease spectrum . There was a linear correlation between serum IgM PG-antibody levels and bacillary index (BI), a measure of bacterial load . Elevated anti-PG IgM in bacillary negative patients was usually indicative of active disease, undetected by BI . We conclude that anti-PG IgM levels are valuable for monitoring the degree of disease activity . Serum anti-PG IgM levels were significantly lower in patients with erythema nodosum leprosum (ENL) as compared to those without ENL, suggesting that IgM PG-antibodies are also involved in the pathogenesis of ENL. Acta Leprol, 1984 Oct-Dec, 2(2-4), 113 - 20 Characterization of Mycobacterium leprae by lipid analysis; Minnikin DE et al.; The lipid composition of the leprosy bacillus, harvested from experimentally infected nine-banded armadillos, strongly supports it status as a distinct species of the genus Mycobacterium . Phthiocerol dimycocerosate waxes and glycosylated phenophthiocerol dimycocerosates are distinct from those characterised from a number of other mycobacteria . The polar lipids of a single isolate lack diacylated forms of phosphatidylinositol di- and pentamannosides, lipids usually found in most mycobacteria . A simple mycolic acid pattern composed of alpha-mycolates and ketomycolates is characteristic of most preparations of M . leprae. Indian J Lepr, 1984 Oct-Dec, 56(4), 776 - 83 Hydrolytic enzymes in macrophages from leprosy patients in presence of Mycobacterium leprae; Marolia J et al.; Presence of Mycobacterium leprae in association with in vitro cultured macrophages, from bacillary negative long term treated lepromatous leprosy patients, induces reduced level of protein and lowering of hydrolytic enzymes like p-glucuronidase, Lysozyme and Lactic dehydrogenase . Alkaline phosphatase, on the other hand is increased . In the macrophages from normal healthy individuals or tuberculoid leprosy patients, presence of M.leprae increases both protein and levels of all the above enzymes . This observation shows that macrophages from lepromatous leprosy patients are unable to manifest in presence of M . leprae, the key enzymes involved in degradation of complex biological entities phagocytosed by the cells. Indian J Lepr, 1984 Oct-Dec, 56(4), 742 - 7 Trophic changes and extensive dissemination in normal mice infected with human Mycobacterium leprae; Vaishnavi C et al.; Closely bred Swiss albino normal mice (Lecca Strain) were inoculated in the footpad with M . leprae, at room temperature . The animals were harvested at 3, 6 and 9 months post inoculation, and bacillary counts were made . Trophic changes were observed in the tail-tips, ears, footpads and forepaws 12-14 months post inoculation in a group which was allowed to survive . The histopathological changes and bacillary infiltration was found in many tissues/organs . The possibility of studying this normal strain of mice as an experimental model for human leprosy has been discussed. Can Med Assoc J, 1984 Oct 1, 131(7), 744 - 8 Adjuvant BCG immunotherapy for malignant melanoma; Paterson AH et al.; A total of 199 patients with stage I malignant melanoma at Clark's level 3 to 5 of invasion were entered into a prospectively controlled randomized clinical trial that attempted to assess the value of local and systemic immunotherapy with BCG (bacille Calmette-Guerin) after surgery . The patients were randomly assigned, with stratification by Clark's level, to receive either routine follow-up or immunotherapy with BCG, administered intradermally with a Heaf gun around the site of wide excision and then given orally for 2 years . Intradermal administration of BCG was repeated after 1 year's oral therapy with BCG . Of the 99 patients in the treatment group 66 had Clark's level 3, 28 had level 4, and 5 had level 5 invasion . Of the 100 patients in the control group, 61 had level 3, 36 had level 4, and 3 had level 5 invasion . Other prognostic factors, such as sex, depth of invasion, histologic features, site of disease and type of surgery, were evenly distributed . There were 57 recurrences of the melanoma, 24 in the treatment group and 33 in the control group . However, this trend was not statistically significant (p = 0.194) . The suggestion that BCG may reduce the likelihood of local/regional recurrence has not been confirmed with longer follow-up . There were 13 such recurrences in the BCG group, compared with 21 in the control group; the proportions of patients in each group who had such a recurrence were not significantly different . Of the 199 patients 41 died, 24 in the control group and 17 in the treatment group; again, this difference was not significant . While there may be minor activity in selected patients, there appeared to be no benefit from this form of adjuvant BCG therapy in patients with malignant melanoma. Transplantation, 1984 Oct, 38(4), 407 - 11 The inhibition of fatal graft-versus-host disease by immunization of donor or host with bacillus Calmette-Guérin cell walls; Janss G et al.; F1 mice receiving sublethal whole-body x-irradiation (300 rads) or treatment with cyclophosphamide prior to the i.p . inoculation of parental spleen cells developed fatal graft-versus-host disease (GVHD) . A greater survival rate was obtained when the inoculated parenteral spleen cells were obtained from BCGcw-preimmunized donors . The immunization of the F1 host with bacillus Calmette-Guerin cell walls (BCGcw) also increased host survival . The combined treatment of preimmunizing the host with BCGcw and of using spleen cells from BCGcw-immunized parental donors to initiate the GVHD resulted in producing the least severe GVHD and the greatest overall survival . The systemic transfer of x-irradiated spleen cells from BCGcw-immunized parental mice inhibited the fatal GVHD induced by the inoculation of normal parental spleen cells . These studies show that BCGcw immunization of the host or obtaining parental spleen cells from BCGcw-immunized animals resulted in improving the overall survival rate in graft-versus-host disease . BCGcw immunization induces suppressor cells and the decrease of graft-versus-host disease that was observed was most likely due to the induction of suppressor cells. Gastroenterology, 1984 Oct, 87(4), 941 - 7 Sarcoidlike granulomas as an early manifestation of Whipple's disease; Cho C et al.; Whipple's disease is often accompanied by a long, preintestinal phase of vague symptoms, such as weight loss, fever, and migratory arthralgia, which may delay diagnosis and proper treatment . We report a patient who presented with sarcoidlike granulomas in the lung 1.5 yr before the development of gastrointestinal symptoms . He was treated with prednisone and his lung lesions improved dramatically . However, steroids could not be discontinued until the diagnosis of Whipple's disease was made and he was started on antibiotic treatment . Whipple's disease was diagnosed from a small intestinal biopsy specimen by electron microscopic demonstration of characteristic bacillary bodies . Liver biopsy specimens also demonstrated a few Kupffer cells containing degenerative bacillary bodies . Based on this case and other reported cases of Whipple's disease with sarcoidlike lesions in various organs, we suggest that sarcoidlike tissue reaction can be an early manifestation of Whipple's disease, recognition of which may have practical value in facilitating an early diagnosis and treatment. DNA, 1984 Oct, 3(5), 347 - 57 Complete nucleotide sequence of a 23S ribosomal RNA gene from Bacillus stearothermophilus; Kop J et al.; A 23S ribosomal (rRNA) gene from Bacillus stearothermophilus has been cloned in pBR322, and its nucleotide sequence determined . The corresponding mature 23S rRNA is predicted to contain 2928 nucleotides . We compare the primary and secondary structures of 23S rRNA from Escherichia coli and B . stearothermophilus, and discuss their potential contributions to thermal stability. J Bacteriol, 1984 Oct, 160(1), 95 - 102 Inverted repeat sequences flank a Bacillus thuringiensis crystal protein gene; Kronstad JW et al.; Two sets of inverted repeat DNA sequences, IR2150 and IR1750, were discovered flanking the crystal protein gene on the 75-kilobase plasmid of Bacillus thuringiensis subsp . kurstaki HD73 . A restriction map of ca . 40 kilobases around the crystal protein gene was constructed, and the positions of the copies of IR2150 and IR1750 were determined . Three copies of IR2150 were found flanking the crystal protein gene in an inverted orientation, and one partial and three intact copies of IR1750 were found in both inverted and direct orientations around the gene . Hybridization experiments with fragments from within IR2150 and IR1750 demonstrated the presence of multiple copies of these sequences on the chromosome of B . thuringiensis subsp . kurstaki HD73 and also revealed a strong correlation between the presence of these sequences and the presence of the crystal protein gene on plasmids from 14 strains of B . thuringiensis. FEBS Lett, 1984 Oct 1, 175(2), 377 - 82 Cloning and expression in Escherichia coli of the insecticidal delta-endotoxin gene of Bacillus thuringiensis var . israelensis; Ward ES et al.; Recombinant plasmids containing the mosquitocidal delta-endotoxin gene were constructed by inserting HindIII fragments of the Bacillus thuringiensis var . israelensis 72-75 Md plasmid in to the Escherichia coli vector pUC12 . Two recombinants producing the 26 000 Da delta-endotoxin (pIP173 and pIP174) were identified by screening clones in an E . coli in vitro transcription-translation system . Both recombinants were 12.4 kb chimaeric plasmids comprising pUC12 and a common 9.7 kb HindIII fragment of the B . thuringiensis plasmid . The 26 000 Da polypeptide synthesis in vivo from pIP174 transformed into E . coli JM101 was lethal to mosquito larvae and cytotoxic to mosquito cells in vitro . The biological authenticity of the cloned product was further confirmed by demonstrating that the cytotoxicity of the polypeptide was neutralised by antiserum to the authentic delta-endotoxin or by preincubation with excess toxin receptor . Transcription of the recombinant delta-endotoxin gene in E . coli appears to utilise a Bacillus promoter sequence(s) rather than the pUC12 beta-galactosidase promotor. J Immunol, 1984 Oct, 133(4), 1735 - 9 Role of the thymus in control of autoreactivity or allotolerance in syngeneic and allogeneic bone marrow chimeras treated with bacterial adjuvants; Taniguchi K et al.; Thy-1-bone marrow (BM) cells from C57BL/6 (B6) mice were transferred into thymectomized or non-thymectomized syngeneic B6----B6, allogeneic B6----C3H or semiallogeneic B6----(B6 X C3H)F1, irradiated mice, after which bacterial substances (bacillus Calmette Guerin {BCG} or Bordetella pertussis {Bp}) were administered within 3 days . The regulation of reactivity toward the host environment, i.e., autoresponsiveness in B6----B6 and allotolerance in B6---C3H, was investigated by monitoring a graft-vs-host (GvH)-like wasting syndrome, as well as the in vitro responsiveness of spleen cells from the reconstituted mice in a mixed leukocyte culture/cell-mediated lysis (MLC/CML) assay . The BCG-treated B6----B6 recipients developed a wasting syndrome and MLC/CML reactivity toward syngeneic target cells within 7 wk . This was never observed in BCG-treated but otherwise normal (i.e., nonreconstituted) mice, nor was it seen in any bone marrow chimeras that had been left without BCG treatment, irrespective of host/donor combination or thymectomy . The development of wasting syndrome as well as autoreactivity in BCG-treated B6----B6 mice could be prevented by thymectomizing the recipients before reconstitution or co-cultivating the donor BM cells with syngeneic spleen cells before reconstitution of nonthymectomized recipients . In the allogeneic or semiallogeneic combinations, the BCG treatment resulted in a wasting syndrome and CML/MLC reactivity toward C3H or (C3H X B6)F1 host-derived cells irrespective of thymic presence or absence . No breakdown of allotolerance, however, was retarded in the thymectomized mice, and it could be prevented by co-cultivation of donor BM cells with splenocytes of recipient genotype only if the cells were used to reconstitute thymectomized recipients . The breakdown of allotolerance in B6----C3H chimera was never accompanied by autoreactivity against B6 target cells . It is concluded that induction of autoreactivity and GvH in BCG-treated syngeneic BM chimeras, probably reflecting the breakdown of autotolerance, is strictly thymus dependent . In contrast, induction of anti-host reactivity in BCG-treated allogeneic chimeras may occur in the absence of a thymus and without concomitant autoreactivity, suggesting two independent levels of controls: one that is thymus dependent for the breakdown of auto- as well as allotolerance, and one that is thymus independent, unique for the breakdown of allotolerance. Med J Aust, 1984 Sep 29, 141(7), 437 - 42 Cephalosporin antibiotic agents; Kemp R; The cephalosporins are a group of antibiotic agents that have been available now for 20 years . Three classes, or generations, of cephalosporins are recognized . The newer third-generation drugs have wider spectra of antibacterial activity; because of this attribute and their ability to achieve high bactericidal titres in CSF these newer compounds constitute an advance in antibiotic therapy by providing safe and effective treatment for Gram-negative bacillary meningitis . The earlier cephalosporins provide cheap, useful and convenient prophylaxis for vascular and orthopaedic operations near the inguinal area . Comparative trials with other broad-spectrum agents need to be performed before the true place of the third-generation agents in anti-infective therapy and prophylaxis is finally determined. J Mol Biol, 1984 Sep 25, 178(3), 743 - 72 Structural evidence for ligand-induced sequential conformational changes in glyceraldehyde 3-phosphate dehydrogenase; Leslie AG et al.; Glyceraldehyde 3-phosphate dehydrogenase is a tetramer of four chemically identical subunits which requires the cofactor nicotinamide adenine dinucleotide (NAD) for activity . The structure of the holo-enzyme from Bacillus stearothermophilus has recently been refined using X-ray data to 2.4 A resolution . This has facilitated the structure determination of both the apo-enzyme and the enzyme with one molecule of NAD bound to the tetramer . These structures have been refined at 4 A resolution using the constrained-restrained parameter structure factor least-squares refinement program CORELS . When combined with individual atomic temperature factors from the holo-enzyme, these refined models give crystallographic R factors of 30.2% and 30.4%, respectively, for data to 3 A resolution . The apo-enzyme has 222 molecular symmetry, and the subunit structure is related to that of the holo-enzyme by an approximate rigid-body rotation of the coenzyme binding domain by 4.3 degrees with respect to the catalytic domains, which form the core of the tetramer . The effect of this rotation is to shield the coenzyme and active site from solvent in the holo-enzyme . In addition to the rigid-body rotation, there is a rearrangement of several residues involved in NAD binding . The structure of the 1 NAD enzyme is asymmetric . The subunit which contains the bound NAD adopts a conformation very similar to that of a holo-enzyme subunit, while the other three unliganded subunits are very similar to the apo-enzyme conformation . This result provides unambiguous evidence for ligand-induced sequential conformational changes in B . stearothermophilus glyceraldehyde 3-phosphate dehydrogenase. Biochem Biophys Res Commun, 1984 Sep 17, 123(2), 831 - 5 Thermal melting of poly(dA-dT).poly(dA-dT) in methanol-water solutions; Kypr J et al.; Thermal melting of a synthetic DNA poly(dA-dT).poly(dA-dT) was investigated in methanol-water solutions . Methanol decreased melting temperature of the polynucleotide but the decrease was qualitatively different in the presence of milimolar concentrations of sodium and cesium cations . The difference is a consequence of the fact that poly(dA-dT).poly(dA-dT) undergoes a helix-helix conformational isomerization in methanol-water solutions which is cesium cation specific . The arising conformation is much more stable than the conformation which poly(dA-dT).poly(dA-dT) adopts in solutions with sodium cations at high methanol concentrations . The (A+T) rich DNA of Bacillus cereus displays a similar behaviour. Eur J Biochem, 1984 Sep 3, 143(2), 359 - 62 Enzymatic preparation of an immunostimulant, the disaccharide-dipeptide, N-acetyl-beta-D-glucosaminyl-(1----4)-N-acetylmuramyl-L-alanyl-D-is ogl utamine, from a bacterial peptidoglycan; Guinand M et al.; The disaccharide-dipeptide N-acetyl-beta-D-glucosaminyl-(1----4)-N-acetylmuramyl-L-alanyl-D-isog lut amine has been obtained by an enzymatic degradation of the peptidoglycan of Actinomadura R39 . The peptidoglycan was hydrolyzed successively by the three following enzymes: lysozyme, DD-carboxypeptidase from Streptomyces albus G and gamma-D-glutamyl-meso-diaminopimelate endopeptidase I from Bacillus sphaericus 9602 . The by-products of the last reaction were eliminated by successive ion-exchange and gel-permeation chromatographies . Both chemical analysis and mass spectrometry show that the resulting disaccharide-dipeptide is a pure compound. Int J Lepr Other Mycobact Dis, 1984 Sep, 52(3), 327 - 30 Serum zinc levels in subtypes of leprosy; Mathur NK et al.; Serum zinc levels were estimated in 146 patients with different types of leprosy and 40 control subjects . Tuberculoid patients were found to have normal serum zinc levels . A gradual reduction in serum zinc levels was observed from TT to LL . Serum zinc levels were not significantly higher in patients treated with dapsone for approximately 18 months . The cause of hypozincemia is not clear, but the findings suggest that there are correlations among bacillary load, immune status, and serum zinc . Zinc deficiency could be one of the factors involved in the nonspecific suppression of cell-mediated immunity in lepromatous leprosy. Onderstepoort J Vet Res, 1984 Sep, 51(3), 155 - 60 The response of Vaal River drift and benthos to Simulium (Diptera: Nematocera) control using Bacillus thuringiensis var . israelensis (H-14); Car M et al.; Two trials to test the efficacy of Bacillus thuringiensis Berliner var . israelensis de Barjac (serotype H-14) against target simuliid and non-target aquatic invertebrates were undertaken in the Vaal River near Warrenton in South Africa . In the 1st trail an application of 1,6 ppm/10 min of B . thuringiensis resulted in a significant (P less than 0,05) reduction of simuliid larvae in rapids 70 m below the treatment point 40 hours after its application . Further downstream the larvicide was ineffectual because the low flow of the river (6 m3/s) allowed the Bacillus pores to settle out in calmer stretches . The 2nd trial was carried out upstream of small rapids with a calculated flow of 0,5 m3/s at a spore concentration of 2,3 ppm/7 min . The effect of B . thuringiensis on the benthic population density and drift activity of the benthos was recorded . A high mortality of simuliid larvae and Ephemeroptera was observed 35 m below the application point 9 hours after the application of the larvicide . The mortality in Ephemeroptera was partially due to the handling of these animals . Population densities of simuliid larvae in the treated rapids decreased 18 hours after application of the larvicide, but small simuliid larvae showed a numerical increase again after 72 hours, indicating rapid recolonization from drifting larvae . Tanytarsine Chironomidae decreased after the application of B . thuringiensis, but most other fauna either increased or did not decrease significantly (P greater than 0,05) . Within 43 minutes after treatment of the rapids with the larvicide, simuliid drift increased more than sixtyfold, revealing the immediate irritating effect of the product on the target organisms.(ABSTRACT TRUNCATED AT 250 WORDS) Mikrobiologiia, 1984 Sep-Oct, 53(5), 796 - 802 {Effect of chloramphenicol and actinomycin D on extracellular alkaline RNAse biosynthesis by Bacillus intermedius}; Znamenskaia LV et al.; By means of chloramphenicol it was found that biosynthesis of alkaline exocellular RNAase was repressed in Bacillus intermedius by inorganic phosphate . Actinomycin D at a low concentration stimulates RNAase biosynthesis in a medium with a minimal phosphorus concentration in model experiments with washed cells and in the batch culture . As a result, the activity of RNAase rises 2-4 times . The stimulating effect of actinomycin D decreases when phosphorus concentration in the medium is increased The effect of actinomycin D is maximal if the antibiotic is added to the medium when the specific growth rate of the bacterium falls down and the rate of RNAase biosynthesis rises. Can J Microbiol, 1984 Sep, 30(9), 1100 - 4 Lectin grouping of Bacillus thuringiensis serovars; DeLucca AJ 2nd; Twelve lectins, 11 of plant and 1 of animal origin, were tested against the 28 serovars of Bacillus thuringiensis to study their agglutinating specificities . Except for the sialic specific lectin from Limulus polyphemus, tube agglutination assays were performed using lectin concentrations of 10, 50, and 100 micrograms against 0.2 mL of 10(9) cells/mL in a final volume of 1.0 mL . The agglutination studies with the Limulus lectin were performed using 10 and 50 micrograms of lectin . Tubes were incubated overnight at room temperature (25 degrees C) and observed for agglutination patterns . Ten of the 28 serovars were individualized according to their ability to bind with various lectins . The study shows that the various serovars have different carbohydrate residues which indicates that the O-somatic antigens differ. Ann Microbiol (Paris), 1984 Sep-Oct, 135B(2), 209 - 17 {Biochemical characterization of Bacillus benzoevorans sp . nov., a new filamentous, sheathed mesophilic species, degrading various aromatic acids and phenols}; Pichinoty F et al.; The eleven strains studied were prototrophic and did not grow in media containing only 1% Bacto-peptone or Bacto-tryptone; they grew rapidly in media containing 0.4% yeast extract and 0.2% sodium acetate or benzoate . The maximal growth temperature ranged from 39 to 45 degrees C . Six aliphatic acids, four aromatic acids and five phenols were used as sole carbon and energy sources by the 11 strains . Carbohydrates and amino acids (except for glycine) were not used as carbon and energy sources . Nitrate (but not nitrite) was used anaerobically as a respiratory electron acceptor . Nitrous oxide was used and reduced to N2 by only 3 strains . The mean guanine-plus-cytosine content of the DNA was 41.3 +/- 1.1 mol % . Morphologically and nutritionally, the bacteria described are clearly different from the 5 known species of the first morphological group whose cells have a diameter greater than 1 micrometer: Bacillus megaterium, B . cereus, B . cereus var . mycoides, B . macroides, B . badius, and B . fastidiosus . Strain B1 (=CCM 3364) is the holotype of Bacillus benzoevorans sp . nov. Ann Microbiol (Paris), 1984 Sep-Oct, 135B(2), 199 - 207 {Morphology and cytology of Bacillus benzoevorans, a new filamentous, sheathed mesophilic species, degrading various aromatic acids and phenols}; Pichinoty F et al.; When cultivated on solid medium or in stationary liquid medium, most of the 11 studied strains of Bacillus benzoevorans grew as unbranched, flexible, immotile filaments (or trichomes) of undefined length and 1.8 micrometer diameter . They were enclosed in a sheath giving an overall diameter of 3.6 micrometer . When cultivated in vigorously shaken liquid medium, several strains grew as separate rods (1.8 X 2.2 micrometer), did not deform the filament and contained dipicolinic acid . The cytoplasm had a granular aspect due to the presence of poly-beta-hydroxybutyric acid . The Gram reaction was variable but the Gram type was positive . Isolated and purified sheath contained 73% proteins, 12.9% reducing sugars and 8% lipids . In stationary liquid medium, the culture had a mycelial aspect and a thick pellicle formed at the surface . Colonies were circular, flat, opaque, whitish, mat and compact; they had irregular edges, spread out on the surface and did not adhere to the agar. Pediatr Infect Dis, 1984 Sep-Oct, 3(5), 420 - 2 A nosocomial common source outbreak caused by Pseudomonas pickettii; Gardner S et al.; Pseudomonas pickettii, a Gram-negative bacillus which has been recovered rarely from clinical specimens, was isolated from the respiratory tracts of 9 of 29 (31%) pediatric intensive care unit patients . The reservoir of the organism was intrinsically contaminated single dose vials of tracheal irrigant solution . Four additional hospitals in three other states have notified the Centers for Disease Control of respiratory colonization with aerobic nonfermentative Gram-negative organisms and have associated this colonization with the use of the same tracheal irrigant solution . Because of the potential for intrinsic contamination, single dose vials must be added to the list of potentially hazardous environmental agents. J Urol, 1984 Sep, 132(3), 457 - 9 Long-term results and complications of intracavitary bacillus Calmette-Guerin therapy for bladder cancer; Morales A; Clinical studies on intracavitary bacillus Calmette-Guerin therapy for bladder cancer have been conducted at this institution for more than 10 years . The 82 patients treated for prophylaxis of multiple superficial recurrences, residual tumors or carcinoma in situ have been followed for 2 to 7 years after treatment . The long-term results confirm previous studies showing the effectiveness of bacillus Calmette-Guerin in the prophylaxis and therapy of superficial vesical neoplasms . However, some decrease is observed in the population free of disease with a prolonged followup . Recent modifications in the original protocol have been introduced to enhance the effectiveness of the vaccine . Side effects during or shortly after treatment were minor and self-limiting in the large majority of patients (fever, bladder irritability and hematuria) . Mild, transient hepatic dysfunction was noted in 2 patients . Hematological disturbances were insignificant . No patient had permanent structural or functional alterations of the bladder. Fed Proc, 1984 Sep, 43(12), 2755 - 9 Regulation of the host response to bacterial lipopolysaccharides; Ulevitch RJ et al.; Bacterial endotoxins or lipopolysaccharides (LPS) are unique glycolipids present in the outer cell membrane of all gram-negative bacteria . It is now generally recognized that LPS is of primary importance in initiating the pathophysiological changes that often accompany gram-negative bacillary infections in humans including hypotensive shock, disseminated intravascular coagulation, and metabolic abnormalities . Although the biochemical mechanisms of these changes are not well understood, increasing emphasis has been placed on defining the biochemical response of the macrophage (M phi) to LPS . In this paper we describe two M phi-derived factors induced by LPS that may be important in the expression of endotoxic activity in the host . These are a procoagulant activity, which is present on the cell membrane of LPS-treated rabbit liver M phi and acts by directly activating coagulation factor X, and a factor released into the supernatant by LPS-treated peritoneal exudate M phi, which suppresses steroidogenesis in explanted adrenocortical cells . The potential role of the M phi in regulating the binding of LPS to high-density lipoproteins through the induction of acute phase proteins is also considered. Biochem Int, 1984 Sep, 9(3), 319 - 25 Induction of deoxyribose-5-phosphate aldolase of Bacillus cereus by deoxyribonucleosides; Tozzi MG et al.; In Bacillus cereus purine ribonucleosides and deoxyribonucleosides share a common inducible catabolic pathway, leading to the formation of ribose-5-P or deoxyribose-5-P respectively inside the cell, while the purine ring remains in the external medium . Both ribo- and deoxyribonucleosides are inducers of adenosine deaminase, inosine-guanosine phosphorylase and phosphopentomutase, the enzymes of the catabolic pathway . We now show that deoxyribonucleosides, but not ribonucleosides, induce the aldolase specific for deoxyribose-5-P (2-deoxy-D-ribose-5-phosphate acetaldehyde lyase, EC 4.1.2.4), thus allowing the sugar moiety of exogenous deoxyribonucleosides to be utilized as an energy source. J Biochem Biophys Methods, 1984 Sep, 9(4), 343 - 50 Deoxyribose 1-phosphate: radioenzymatic and spectrophotometric assays; Ipata PL et al.; A method has been developed to measure deoxyribose 1-phosphate in the presence of ribose 1-phosphate and other sugar phosphates . The specificity of the method is based on the observation that only deoxyribose 1-phosphate is hydrolyzed by heating at pH 7.4, while both deoxyribose 1-phosphate and ribose 1-phosphate remain unchanged when heated at pH 10 . A tissue extract is heated at pH 10 . The amount of deoxyribose 1-phosphate plus ribose 1-phosphate is determined from that of deoxyinosine plus inosine formed in a coupled enzymatic reaction, based on the following two-stage transformation: deoxyribose 1-phosphate (ribose 1-phosphate) + adenine in equilibrium deoxyadenosine (adenosine) + inorganic phosphate, catalyzed by adenosine phosphorylase; deoxyadenosine (adenosine) + H2O----deoxyinosine (inosine), catalyzed by adenosine deaminase . By taking advantage of its unique heat lability, deoxyribose 1-phosphate is eliminated by heating the tissue extract at pH 7.4, and ribose 1-phosphate is determined as above . The amount of deoxyribose 1-phosphate stems from the difference between the amount of deoxyinosine plus inosine measured in the tissue extract heated at pH 10 and that of inosine measured in the tissue extract heated at pH 7.4 . Free deoxyribose 1-phosphate has been found in rat tissues, as well as in Bacillus cereus during stationary phase of growth. J Bacteriol, 1984 Sep, 159(3), 820 - 4 Bacillus cereus electron transport and proton motive force during aerotaxis; Laszlo DJ et al.; Aerotaxis (migration towards oxygen) of Bacillus cereus M63, a motile strain, was inhibited by potassium cyanide and 2-heptyl-4-hydroxyquinoline N-oxide, indicating a requirement for both the terminal oxidase (cytochrome aa3) and the cytochrome b segment of the electron transport system . The concentration of oxygen that gave a half-maximal aerotactic response (K0.5) was 0.31 microM, which was similar to the Km for respiration (0.80 microM) . The proton motive force increased from -135 to -177 mV when anaerobic cells were aerated, and it is proposed that the signal for aerotaxis is the increase in proton motive force that results from increased respiration . A strain of B . cereus T initially used in this study was immotile, grew as long chains of cells, and was deficient in autolytic enzyme . B . cereus M63 is a spontaneous derivative of B . cereus T that has normal motility. Biochem J, 1984 Sep 1, 222(2), 389 - 94 Effect of phospholipase C (Bacillus cereus) on freshly isolated and 4-day-stored human platelets; Solberg C et al.; Phospholipase C (from Bacillus cereus) was used to study fresh and stored human platelets . Provided that the enzyme was inactivated before lipid extraction, no significant degradation of phospholipid in fresh cells was noted, even when platelets were activated or induced to change shape by ADP, collagen or thrombin . With platelets isolated from concentrates stored for transfusion for 4 days at 22 degrees C, membrane phospholipids were degraded by the enzyme to an extent depending on the pH in the platelet concentrate at day 4 of storage . The extent of phospholipid hydrolysis in platelets correlated well with the extent of release of lactate dehydrogenase during storage, with both being minimal for platelets from concentrates of final pH 6.5-6.9 . Under non-lytic conditions, phosphatidylcholine was the phospholipid most degraded (40%), with no significant degradation of phosphatidylserine being detected . Storage does not seem to alter the distribution of phospholipids at the external leaflet of the plasma membrane. Jpn J Surg, 1984 Sep, 14(5), 413 - 9 Eradication of microscopic metastases with intratumoral injection of bacillus Calmette-Guerin; Sakita M et al.; The studies reported here were designed to examine the effects of intratumoral preoperative administration of Bacillus Calmette-Guerin (BCG) on the cure rates of C3H mice transplanted with MH134 tumor cells and on the metastatic rates in the regional lymph nodes . Furthermore, the morphological findings occurring in the regional lymph nodes were monitored during tumor growth using H-E stain and non-specific esterase staining . The cure rate of the Group treated with BCG intratumoral injection and surgery was significantly higher than that of the Group treated with surgery alone, and in the BCG + surgery group metastatic rates of regional lymph nodes decreased consistently after operation . Moreover, in this group, extensive sinus histiocytosis and marked swelling of the regional nodes were frequently observed . Quantitative studies of the cell kinds using the esterase staining indicated that intratumoral injection of BCG has an effect on the influx of lymphoid cells into the regional nodes, but does not aid specific cell lineage to flow into the regional nodes . In cytostatic assays, it was shown that the regional lymph node cells and spleen cells in the BCG + surgery group always have a greater per cent of inhibition than those in the surgery alone group. J Biochem (Tokyo), 1984 Sep, 96(3), 701 - 11 A comparative study of sulfhydryl groups required for the catalytic activity of gramicidin S synthetase and isoleucyl tRNA synthetase; Kanda M et al.; The sulfhydryl groups required for the catalytic activity of gramicidin S synthetase of Bacillus brevis and Escherichia coli isoleucyl tRNA synthetase were compared . In gramicidin S synthetase 2(GS 2), about four sulfhydryl groups react rapidly with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) or N-ethylmaleimide (NEM), and are essential for gramicidin S formation in the presence of gramicidin S synthetase 1 (GS 1) . These sulfhydryl groups are protected against DTNB and NEM reactions by the preincubation of GS 2 with amino acid substrates in the presence of ATP and MgCl2, like the sulfhydryl groups that react rapidly with DTNB or NEM and are required for the catalytic activity of GS 1 and isoleucyl tRNA synthetase . In GS 2, GS 1, and isoleucyl tRNA synthetase, the sulfhydryl group that reacts rapidly with NEM and is required for the catalytic activity is involved in the amino acid binding as a thioester . In isoleucyl tRNA synthetase, it is suggested that isoleucine may be transferred from the isoleucine thioester enzyme complex to tRNA by a mechanism similar to that proposed for gramicidin S synthetase. J Leukoc Biol, 1984 Sep, 36(3), 293 - 306 Differential expression of macrophage effector functions: bactericidal versus tumoricidal activities; Campbell PA et al.; Macrophage populations may be induced to express tumoricidal or bactericidal activities following exposure to certain stimuli . An understanding of the differences in the stimulatory mechanisms and in the characteristics of the macrophages they affect will be facilitated by comparing functional activities of various macrophage populations . The experiments described here were conducted to determine whether injection of a single stimulus necessarily drives cells to express both tumoricidal and bactericidal activities or whether selected reagents can drive cells to express one activity without expressing the other . The data show that a single population of mouse or hamster peritoneal exudate cells obtained following injection of proteose peptone is bactericidal for Listeria monocytogenes and for E . coli, but is not tumoricidal for TCMK-1, Ad2HE3, or mKS-A TU-5 target cells . In contrast, peritoneal exudate cells collected after injection of Bacillus Calmette Guerin (BCG) organisms are always highly tumoricidal, and either show no effect on Listeria monocytogenes or E . coli, or are at best bacteriostatic . Data indicate that the effector cells in these assays are macrophages, that the dissociation of tumoricidal and bactericidal activity occurs over a wide dose range, and that the tumoricidal capabilities are not artifacts of the assay system . These results suggest that a given macrophage population may preferentially express tumoricidal or bactericidal activities depending on the stimulus used. Am J Med, 1984 Sep, 77(3), 551 - 4 Infection of a ventricular aneurysm and cardiac mural thrombus . Survival after surgical resection; Venezio FR et al.; Infections of cardiac mural thrombi are rare, and because antemortem diagnosis is difficult and antibiotic therapy alone ineffective, the associated mortality has been significant . A patient with gram-negative bacillary infection of a mural thrombus is described . Gallium 67 citrate isotope scanning and two-dimensional echocardiography were helpful adjuncts in establishing the diagnosis . Surgical resection of the infected myocardial tissue and prolonged antimicrobial therapy were necessary for cure. J Urol, 1984 Sep, 132(3), 570 - 3 Long term protection in bladder cancer following intralesional immunotherapy; Reichert DF et al.; Despite effective treatment of existing tumors, patients with bladder cancer remain at risk of developing new tumors . Effective immunotherapy may lower that risk . To test this hypothesis, mice that had survived transitional cell carcinoma (MBT2) transplantation with the aid of bacillus Calmette-Guerin immunotherapy were randomized and tested for long term protective immunity against bladder carcinoma . Fifty-one tumor-free mice that had survived tumor challenge 10 to 15 months previously were randomized into 3 groups to receive intradermal tumor .noculation and intraperitoneal levamisole, intralesional Tice strain bacillus Calmette-Guerin, or intralesional saline . Fifteen previously unchallenged animals also received tumor and intralesional saline . All 3 groups of survivors had less tumor growth (p less than 0.01) than nonsurviving controls . Even among survivors, additional bacillus Calmette-Guerin immunization, but not levamisole treatment, significantly inhibited tumor growth (p less than 0.01) . A 2nd experiment compared 22 nonimmune mice, 21 mice preimmunized intravenously with 300 micrograms of bacillus Calmette-Guerin cell walls, and 18 mice that had survived MBT2 by 8 months after live bacillus Calmette-Guerin treatment . Nonimmune and survivor groups were randomly subdivided into saline or treatment groups . Cell wall-preimmunized mice were divided into matching groups according to footpad response to purified protein derivative . The cell-wall preimmunized and nonimmune mice received the immunostimulant P3+Re-glycolipid or the carrier solution alone . The group of survivors received either intralesional saline or live bacillus Calmette-Guerin . Both bacillus Calmette-Guerin and saline-treated groups had significantly less tumor growth (p less than 0.001) than nonsurviving controls . Animals treated with P3-Re-glycolipid (with or without preimmunization with cell wall) did not differ from nonsurviving control . Footpad response to purified protein derivative did not correlate with tumor growth in these mice . Our results suggest that intralesional bacillus Calmette-Guerin immunotherapy can afford long term protection from transplanted bladder cancer, and that live bacillus Calmette-Guerin is superior to levamisole and P3 + Re-glycolipid + bacillus Calmette-Guerin cell walls in the treatment of bladder cancer. Cell Immunol, 1984 Sep, 87(2), 553 - 65 Lysis of mouse macrophages, fibroblasts, and epidermal cells by epidermal alloantigen-specific cytotoxic T lymphocytes: effect of culture and inflammatory agents on Epa-1 expression; Burlingham WJ et al.; The expression of Epa-1, a tissue-restricted non-major histocompatibility complex (MHC) alloantigen, on CBA epidermal cells (EC), fibroblasts (FB), and macrophages (M phi) was investigated using bulk-cultured and clonally-derived anti-Epa-1 cytotoxic T lymphocytes (CTL) . Epa-1 was readily detected on freshly trypsinized and 24-hr-cultured EC, and on skin FB cul