Can J Microbiol, 1978 Nov, 24(11), 1289 - 95 Production and characterization of two hemolysins of Bacillus cereus; Coolbaugh JC et al.; Bacillus cereus strain B-48 produced two hemolysins with molecular weights of 52,000 (H-I) and 31,000 (H-II) . A mutant was isolated that produced only H-II but was identical with the wild type in all other respects . We exploited this mutant to produce H-II for study that was free of contamination by H-I . By manipulation of media composition, we produced H-I in the absence of H-II . The hemolysins were precipitated differently by ammonium sulfate, and both exhibited the Arrhenius effect when heated . Both hemolysins attached rapidly to erythrocytes; however, lysis by H-I was immediate, while lysis by H-II followed after a lag . Hemolysis by H-I and H-II increased in rate with increasing temperature and was absent at 0 degrees C . Only H-I was inhibited by cholesterol . The hemolysins of B . cereus appeared similar to the hemolysins of B . thuringiensis . H-I probably is identical with cereolysin. Chin Med J (Engl), 1978 Nov, 4(6), 497 - 500 Experience in emergency treatment of shock due to infection; Wang ST et al.; PIP: 188 cases of shock due to infection were treated at the People's Hospital in Peking between 1964-1965 and 1972-1974 . 133 (70.7%) of the cases were due to pneumonia or toxic bacillary dysentary . 71 of the 133 cases were treated with vasoconstrictors and 62 with vasodilators . Those treated with vasoconstrictors had a mortality rate of 16.9% while the 62 treated with vasodilators were cured . Average duration of treatment was shorter with the vasodilator group (23.6 hours in 52 cases) than vasoconstrictors (54.6 hours in 48 cases) . As a result of the study it is now believed that hypotension in shock is due to relative or absolute lack of effective circulatory blood volume as a result of microcirculatory impairment . Rather than focusing solely on maintenance of blood pressure primary emergency measures for shock should focus on restoring blood perfusion of body tissue . Comments were made regarding blood volume, selection of vasoactive agents with anisodamine the preferred drug and prevention of acute renal failure and pulmonary endema . Zentralbl Bakteriol {Orig A}, 1978 Nov, 242(1), 137 - 40 Induced chain formation in Bacillus megaterium by suramin (Bayer 205); Shaikh D et al.; B . megaterium NCTC 5637 has been shown to grow into long chains in 1% glucose and 1% suramin (w/v) . This effect was not noticeable in lower concentration of the inhibitor . The effect diminished in suramin free medium, indicating the change to be phenotypic. Cancer Treat Rep, 1978 Nov, 62(11), 1613 - 21 Pharmacologic factors and manipulation of immunity systemic adjuvants in cancer therapy; Mathe G et al.; Because of the experimental and clinical studies which have been extensively conducted with bacillus Calmette-Guerin (BCG) as a systemic adjuvant in cancer immunotherapy, we have analyzed the main factors and conditions which determine its beneficial action and have underlined some of these (eg, the dose factor which controls the amplification of suppressor cells which is probably responsible for failures and even the possible tumor-enhancing effect of immunotherapy) . Knowing those factors and conditions, we have been able to establish a systematic immunopharmacologic study of systemic immunity adjuvants, which has resulted in the discovery of agents whose actions are more rapid than that of BCG on one or a few populations of cells involved in immunity and which, unlike BCG, do not induce suppressor cell amplification . This amplification may explain the difference in the results obtained with this mycobacterium in various clinical immunotherapy trials in which it was applied differently . It is proposed to combine these mono- or pauc-functional adjuvants in order to try to obtain all of the beneficial effects of BCG without the amplification of suppressor cells. J Antibiot (Tokyo), 1978 Nov, 31(11), 1162 - 9 Clavulanic acid inhibition of beta-lactamase I from Bacillus cereus 569/H; Durkin JP et al.; Inactivation of beta-lactamase I by clavulanic acid was investigated . Clavulanic acid induced inhibition of the enzyme was found to be progressive with time . Benzylpenicillin provided protection against the adverse effects of the inhibitor initially, however, the enzyme was irreversibly inhibited in a progressive manner even in the presence of substrate . Reaction of beta-lactamase I with clavulanic acid, in the presence of ampicillin, led to a very rapid inactivation of the enzyme. Am J Med, 1978 Nov, 65(5), 873 - 80 Whipple's disease of the lung; Winberg CD et al.; Described here is a unique case of Whipple's disease in a 54 year old man with chronic severe cough and gastrointestinal symptoms in whom the initial diagnosis of Whipple's disease was made by lung biopsy . This is, to our knowledge, the first reported case in which the bacilliform structures of Whipple's disease have been demonstrated in tissues from other than the gastrointestinal tract of lymph nodes . Subsequently, a peroral biopsy of the small intestine was performed and revealed identical and pathognomonic features of Whipple's disease . The pulmonary roentgenologic findings are described and the histologic differential diagnosis of histiocytic infiltrates in the lung, which may be histologically similar to Whipple's disease, are briefly reviewed. Biochem J, 1978 Nov 1, 175(2), 441 - 7 Histidine residues of zinc ligands in beta-lactamase II; Baldwin GS et al.; 1 . The Zn(II)-requiring beta-lactamase from Bacillus cereus 569/H/9, which has two zinc-binding sites, was examined by 270 MHz 1H n.m.r . spectroscopy . Resonances were assigned to five histidine residues . 2 . Resonances attributed to three of the histidine residues in the apoenzyme shift on the addition of one equivalent of Zn(II) . 3 . Although these three histidine residues are free to titrate in the apoenzyme, none of them titrates over the pH range 6.0--9.0 in the mono-zinc enzyme . 4 . The ability of the C-2 protons of these three histidine residues to exchange with solvent (2H2O) is markedly decreased on Zn(II) binding . 5 . It is proposed that these three histidine residues act as zinc ligands at the tighter zinc-binding site . 6 . Resonances attributed to a fourth histidine residue shift on addition of further zinc to the mono-zinc enzyme . It is proposed that this histidine residue acts as a Zn(II) ligand at the second zinc-binding site. J Biol Chem, 1978 Oct 10, 253(19), 6738 - 43 Membrane bioenergetic parameters in uncoupler-resistant mutants of Bacillus megaterium; Decker SJ et al.; Mutants of Bacillus megaterium displaying malate-driven ATP synthesis resistant to uncouplers of oxidative posphorylation are further characterized . Both the pH gradient and electrical potential generated across the membrane by malate respiration are equally sensitive to uncouplers in the wild type and uncoupler-resistant mutants . The mutants possess 0 to 10% of the wild type ATPase activity which is not activated by pretreatment with heat or trypsin . Despite this inability to measure ATPase activity, the mutants demonstrate acid-pulse-driven ATPase synthesis which is sensitive to uncouplers as well as malate-driven ATP synthesis which becomes uncoupler sensitive at pH 5.5 . N,N' -Dicyclohexylcarbodiimide and valinomycin plus potassium inhibition of ATP synthesis is reversed by uncouplers in the mutants but not in the wild type . The data support the existence of a specific site on the ATPase complex for uncoupler binding which, if altered by mutation, affects uncoupler binding to the complex . The retention of malate-driven ATP synthesis in the absence of a significant pH gradient or electrical potential suggests that an alternative intermediate is involved in coupling oxidation to phosphorylation. Arch Dermatol, 1978 Oct, 114(10), 1501 - 4 Combined immunotherapy of malignant melanoma . Unusual survival following cerebral metastasis; Spitler LE et al.; An 18-year-old woman was found to have solitary cerebral, choroidal, and pulmonary metastases of malignant melanoma three years after excision of a primary malignant melanoma . The cerebral metastasis was excised, and the patient's condition was treated with CNS irradiation followed by combined immunotherapy with transfer factor and Bacille bilie de Calmette-Guerin . The transfer factor donor was her father, who showed cellular immunity to melanoma extracts on in vitro testing . Histologic examination of the pulmonary nodule, which was excised after the initiation of immunotherapy, revealed a dense lymphocytic infiltrate associated with the metastatic melanoma . The patient is currently free of detectable melanoma more than three years after the cerebral metastasis . Studies in a second patient also demonstrated the appearance of inflammatory infiltrate in metastatic melanoma following transfer factor therapy. J Bacteriol, 1978 Oct, 136(1), 24 - 34 Modulation of an apparent mRNA pool for extracellular protease in Bacillus amyloliquefaciens; O'Connor R et al.; Late-log-phase cells of Bacillus amyloliquefaciens have the unusual capacity to produce extracellular protease for over 60 min in the presence of rifampin or actinomycin D at levels which strongly inhibit incorporation of amino acids into cellular protein . If cells are incubated in the presence of high levels of amino acids for 75 min this capacity is exhausted, but it is retained if the incubation is carried out in low levels of amino acids . Transfer of exhausted cells from high to low concentrations of amino acids results in a progressive recovery of the capacity for rifampin-actinomycin-insensitive protease production . The results seem best explained on the basis of the accumulation of a reserve pool of mRNA for extracellular protease . Measurement of the apparent mRNA pool size over 12 h shows a cyclical rise and fall, and these changes correlate with a periodic variation of the rate of protease production . A working hypothesis is presented to account for these observations in terms of a novel control situation over protease mRNA transcription. Appl Environ Microbiol, 1978 Oct, 36(4), 625 - 6 Purification of the protein crystal from Bacillus thuringiensis by zonal gradient centrifugation; Ang BJ et al.; A method is described for the large-scale purification of the Bacillus thuringiensis protein crystal by zonal gradient centrifugation . NaBr gradients are employed in a Beckman J21-B centrifuge equipped with a JCF-Z rotor. Cancer Res, 1978 Oct, 38(10), 3150 - 3 Controlled trial of methotrexate and Bacillus Calmette-Guérin therapy for advanced head and neck cancer; Papac R et al.; Thirty-eight patients with advanced, inoperable squamous cell carcinoma of the head and neck were randomized to receive methotrexate alone or methotrexate with Bacillus Calmette-Guerin . The response rates with methotrexate (3 of 19) and methotrexate plus B . Calmette-Guerin (4 of 16) were similar, as was the duration of response and survival of the two groups . The results of in vitro immunological studies of lymphocytes were assessed . Marked weight loss, poor performance status, and distant metastases were the most important prognostic factors . The presence of anergy was significantly correlated with weight loss . This study also indicated that a large tumor burden is a frequent occurrence in advanced head and neck cancer and may account for the lack of efficacy of B . Calmette-Guerin. Proc Natl Acad Sci U S A, 1978 Oct, 75(10), 5142 - 4 Systemic bacillus Calmette-Guérin (BCG) activates natural suppressor cells; Bennett JA et al.; Addition of normal C57BL/6 mouse bone marrow cells to an in vitro culture of normal C57BL/6 spleen cells and allogeneic P815-Y tumor cells inhibited the development of cell-mediated immunity . Bacillus Calmette-Guerin (BCG) enhanced the suppressive activity of these bone marrow cells as early as 2 days after its intravenous administration to donor mice and elicited similar activity in the spleen by 7 days . Concomitant with the appearance of suppressor cells in the spleen there was a decrease in bone cell number and an increase in spleen cell number . While normal spleen cells failed to inhibit immunization, spleen cells from thymectomized, irradiated, bone marrow-reconstituted mice were inhibitory . Administration of BCG further increased the suppressive activity of spleen cells in these T cell-deprived mice . From this evidence it appears that systemic administration of BCG activates natural suppressor cells in the bone marrow and elicits suppressor cells in the spleen through the migration and colonization of the spleen by bone marrow elements. J Bacteriol, 1978 Oct, 136(1), 433 - 6 Levels of cyclic GMP in dormant, germinated, and outgrowing spores and growing and sporulating cells of Bacillus megaterium; Setlow B et al.; The level of cyclic GMP was less than one molecule per organism in dormant, germinated, and outgrowing spores of Bacillus megaterium . A significant level (approximately 8 pmol/g, dry weight) of cyclic GMP was found in early to mid-log phase cells, but the level fell to below 0.2 pmol/g, dry weight, in late-log phase and only rose slightly to approximately 0.9 pmol/g, dry weight, in stationary phare . No significant amount of cyclic GMP was detected in the growth medium at any time. Can J Microbiol, 1978 Oct, 24(10), 1227 - 35 Chromosome age and segregation during sporulation of Bacillus megaterium; Hitchins AD; The effect of chromosome age on segregation during sporulation was investigated . Vegetative cells of Bacillus megaterium were labeled with {Me-3H}thymine and then were grown at 30 degrees C in nonradioactive medium for various times before being allowed to sporulate . The ratio of the amount of label in sporal DNA to that in sporangial DNA, obtained after minor correction for the sporulation frequency, remained essentially constant as the postlabeling growth period was increased from one to seven generations . The spores were preferentially located at the older poles of sporangia, i.e . the poles formed by divisions occurring prior to those forming the sporangia . Therefore, it seems that old (labeled) chromosomes segregate randomly with respect to both the morphological and genealogical polarities of sporangia . Examination of total cell lysates by dye-buoyant density gradient centrifugation revealed the presence of covalently closed circular DNA from cells grown at 37 degrees C, but none was obtained from cells grown at 30 degrees C . Thus, possible interference by large amounts of extrachromosomal DNA in the determination of the chromosomal segregation pattern is unlikely. Ann Ophthalmol, 1978 Oct, 10(10), 1367 - 70 Endogenous endophthalmitis associated with bacillus cereus bacteremia in a cocaine addict; Masi RJ; A 22-year-old black female intravenous cocaine addict presented with an endophthalmitis of the right eye . Diagnostic evaluation included an immediate anterior chamber paracentesis and a delayed vitreous aspiration . Although cultures from the involved eye were negative, all 7 blood cultures grew Bacillus cereus suggesting that this organism was the responsible agent of an endogenous endophthalmitis . The patient was treated with appropriate systemic and local antibiotics with resolution of the acute inflammatory signs . However, a phthisical eye has been noted on follow-up examinations. J Bacteriol, 1978 Oct, 136(1), 331 - 40 Purification and characterization of additional low-molecular-weight basic proteins degraded during germination of Bacillus megaterium spores; Setlow P; Dormant spores Bacillus megaterium contained a group of low-molecular-weight (5,000 to 11,000) basic (pI greater than 9.4) proteins (termed D, E, F, and G proteins) which could be extracted from disrupted spores with strong acids . These proteins were distinct from the previously described A, B, and C proteins which are degraded during spore germination . However, the D, E, F, and G proteins were also rapidly degraded during spore germination, accounting for 10 to 15% of the protein degraded . Proteins similar to the D, E, F, and G species were also present in spores of other bacterial species . In B . megaterium, the D, E, F, and G proteins were low or absent (less than 15% of the spore level) in vegetative and young sporulating cells and appeared only late in sporulation . The D, E, F, and G proteins were purified to homogeneity, and all contained a high percentage of hydrophilic amino acids; one protein (G) contained 31% basic amino acids and also contained tryptophan . All four proteins were rapidly degraded in vitro by dormant spore extracts . Two proteins (D and F) were degraded in vitro by the previously described spore protease which initiates degradation of the A, B, and C proteins in vivo; the spore enzyme (s) degrading proteins E and G have not been identified. J Bacteriol, 1978 Oct, 136(1), 209 - 18 Isolation of stable ribosomal subunits from spores of Bacillus cereus; Kieras RM et al.; Analyses of ribosomes extracted from spores of Bacillus cereus T by a dryspore disruption technique indicated that previously reported defects in ribosomes from spores may arise during the ribosome extraction process . The population of ribosomes from spores is shown to cotain a variable quantity of free 50S subunits which are unstable, giving rise to slowly sedimenting particles in low-Mg2+ sucrose gradients and showing extremely low activity in in vitro protein synthesis . The majority of the ribosomal subunits in spores, obtained by dissociation of 70S ribosomes and polysomes, are shown to be as stable as subunits from vegetative cells, though the activity of spore polysomes was lower than that of vegetative ribosomes . In spite of the instability and inactivity of a fraction of the spore's ribosomal subunits, the activity of the total population obtained from spores by the dry disruption technique was 32% of vegetative ribosome activity, fivefold higher than previously obtained with this species . The improvement in activity and the observed variability of subunit destabilization are taken as evidence for partial degradation of spore ribosomes during extraction. Immunology, 1978 Oct, 35(4), 573 - 9 Adjuvant effect of a peptidoglycan attached covalently to a synthetic antigen provoking anti-phage antibodies; Langbeheim H et al.; The synthetic antigen denoted P2-A--L, comprising the fragment P2 of the coat protein of MS-2 coliphage attached to multichain poly-DL-alanine, served for the immunization of guinea-pigs . Immunization was carried out either in phosphate buffered saline (PBS) or in Freund's incomplete adjuvant (FIA) in the presence or absence of a small molecular weight peptidoglycan prepared from Bacillus megaterium, which was checked for its adjuvant effect . The various antisera were assessed by their capacity to neutralize MS-2 bacteriophage viability . When injected in PBS or FIA, P2-A--L did not elicit any measurable anti-phage activity . Addition of the peptidoglycan by simple mixing did not bring about a significant increase in antibody production . However, when the peptidoglycan was chemically linked to the P2-A--L conjugate, it had a marked adjuvant effect when the material was administered in FIA, almost identical to the extent of the effect of Freund's complete adjuvant. Appl Environ Microbiol, 1978 Oct, 36(4), 549 - 51 Effect of hydrostatic tensile stress on the growth of Escherichia coli and Bacillus cereus; O'Brien WJ et al.; The specific growth rates of Escherichia coli and Bacillus cereus were measured for growth media in a flask, a lens-plate arrangement simulating an isolated capillary space, and a lens-plate arrangement under hydrostatic tensile stress . The specific growth rates of the bacteria were the same for the flask and lens-plate arrangement without hydrostatic tensile stress, but were enhanced when the growth media were subjected to hydrostatic tensile stress . The enhanced specific growth rates reached steady values at a tensile stress of 40 pascals . The effect was observed up to tensile stresses of around 100 pascals . The maximum increase in specific growth rate was 25% for E . coli and 22% for B . cereus. Appl Environ Microbiol, 1978 Oct, 36(4), 539 - 43 Microbial catabolism of vanillate: decarboxylation to guaiacol; Crawford RL et al.; A novel catabolic transformation of vanillic acid (4-hydroxy-3-methoxybenzoic acid) by microorganisms is reported . Several strains of Bacillus megaterium and a strain of Streptomyces are shown to convert vanillate to guaiacol (o-methoxyphenol) and CO2 by nonoxidative decarboxylation . Use of a modified most-probable-number procedure shows that numerous soils contain countable numbers (10(1) to 10(2) organisms per g of dry soil) of aerobic sporeformers able to convert vanillate to guaiacol . Conversion of vanillate to guaiacol by the microfloras of most-probable-number replicates was used as the criterion for scoring replicates positive or negative . Guaiacol was detected by thin-layer chromatography . These results indicate that the classic separations of catabolic pathways leading to specific ring-fashion substrates such as protocatechuate and catechol are often interconnectable by single enzymatic transformations, usually a decarboxylation. J Antibiot (Tokyo), 1978 Oct, 31(10), 966 - 9 Ribostamycin production by a mutant of butirosin producing bacteria; Fujiwara T et al.; By the use of our improved colony selection technique, xylostasin and ribostamycin producing mutants were isolated from nitrosoguanidine treated Bacillus circulans B15M, a producer of butirosins A and B . Among these structurally related aminoglycosides, ribostamycin is the well-known product of a Steptomyces and has not been isolated as a bacterial metabolite . A selected mutant of strain 306, which produces xylostasin and ribostamycin, was futher mutagenized in expectation of getting an improved strain having the ability to accumulate a large amount of ribostamycin in the culture broth . One mutant, strain 451, derived from strain 306, produced ribostamycin free of xylostasin. J Antibiot (Tokyo), 1978 Oct, 31(10), 1023 - 30 Mutational biosynthesis of butirosin analogs . I . Conversion of neamine analogs into butirosin analogs by mutants of Bacillus circulans; Takeda D et al.; By N-methyl-N'-nitro-N-nitrosoguanidine treatment, neamine-negative mutants which required neamine for biosynthesis of butirosins were obtained from a butirosin-producing organism Bacillus circulans . These mutants also produced butirosins from paromamine and could be divided into two types I and II . Mutants of type I could not produce butirosins from 2-deoxystreptamine, whereas those of type II could . Two typical mutants MCRL 5003 (type I) and MCRL 5004 (type II) could produce butirosin analogs, 3', 4'-dideoxybutirosins, 6'-N-methylbutirosins, 3', 4'-dideoxy-6'-N-methylbutirosins and 3', 4'-dideoxy-6'-C-methyl-butirosins from neamine analogs, gentamine Cla, 6'-N-methylneamine, 6'-N-methylgentamine Cla and gentamine C2, respectively. Can J Microbiol, 1978 Oct, 24(10), 1164 - 72 Production and properties of polygalacturonate lyase by an alkalophilic microorganism Bacillus sp . RK9; Kelly CT et al.; Bacillus sp . RK9 was isolated from soil and produced a constitutive polygalacturonate lyase . Production of the enzyme required the presence of complex nitrogen (peptone and yeast extract) . Highest activity was obtained with an initial pH of 9.7 . The organism was alkalophilic . No growth occurred below pH 7.5 . The enzyme was purified by salt precipitation and diethylaminoethyl (DEAE) cellulose ion-exchange chromatography . The pH optimum for activity was 10.0 in 0.01 M glycine-NaOH buffer . Calcium alone, of divalent cations, activated the enzyme by 2.9-fold . Complete inhibition of enzyme activity was achieved by 1 mM ethylenediaminetetraacetic acid (EDTA) . Hydrolysis of substrate occurred in a random fashion and the enzyme was 50% more active towards acid soluble pectic acid (ASPA) than towards sodium polypectate. Biochim Biophys Acta, 1978 Sep 26, 536(1), 172 - 83 Nitrogenase from Bacillus polymyxa . Purification and properties of the component proteins; Emerich DW et al.; A purification procedure is described for the components of Bacillus polymyxa nitrogenase . The procedure requires the removal of interfering mucopolysaccharides before the two nitrogenase proteins can be purified by the methods used with other nitrogenase components . The highest specific activities obtained were 2750 nmol C2H4 formed . min-1 . mg-1 MoFe protein and 2521 nmol C2H4 formed . min-1 . mg-1 Fe protein . The MoFe protein has a molecular weight of 215 000 and contains 2 molybdenum atoms, 33 iron atoms and 21 atoms of acid-labile sulfur per protein molecule . The Fe protein contains 3.2 iron atoms and 3.6 acid-labile sulfur atoms per molecule of 55 500 molecular weight . Each Fe protein binds two ATP molecules . The EPR spectra are similar to those of other nitrogenase proteins . MgATP changes the EPR of the Fe protein from a rhombic to an axial-type signal. J Biol Chem, 1978 Sep 25, 253(18), 6516 - 22 Formation and function of N-acetyloglucosamine-linked phosphoryl- and pyrophosphorylundecaprenols in membranes from Bacillus cereus; Yamamori S et al.; Membranes from Bacillus cereus AHU 1356 incorporated radioactivity from UDP-N-acetyl{14C}glucosamine into three alkaline-stable and acid-labile lipids which were extracted into chloroform:methanol (2:1) and separated from each other by thin layer chromatography on silica gel plates . The major labeled lipid (Lipid 1) and a minor one (Lipid 2) were identified as N-actetylglucosaminyl phosphorylundecaprenol from several analytical criteria involving mass spectral data and from reversal of their formation by UDP . These two lipids appear to differ in geometry of their polyprenol moieties . The third labeled lipid (Lipid 3) was identified as N-acetylglucosaminyl pyrophosphorylundecaprenol . Antibiotic 24010, a tunicamycin-like antibiotic, at 1 microgram/ml was found to inhibit almost completely the formation of Lipid 3, whereas it inhibited the formation of Lipid 1 much more weakly and rather enhanced the formation of Lipid 2 . Radioactivity was also incorporated into a polymer from UDP-GlcNAc and from Lipid 3 . UDP-N-acetylmannosamine, UDP-N-acetylgalactosamine, and UDP-glucose supported the incorporation . Antibiotic 24010 strongly inhibited the incorporation of radioactivity from UDP-GlcNAc into polymer, whereas it did not affect the incorporation from Lipid 3 . Thus, it is concluded that N-acetylglucosaminyl pyrophosphorylundecaprenol serves as a precursor in the synthesis of a polymer presumed as the cell wall polysaccharide of this bacterial strain. J Biol Chem, 1978 Sep 10, 253(17), 5899 - 901 Unusual COOH-terminal structure of staphylococcal protease; Drapeau GR; The extracellular enzyme, staphylococcal protease, carries a COOH-terminal tryptic peptide of 43 amino acid residues most of which are aspartic acid, asparagine, and proline . This peptide might have a function equivalent to that of a similar segment previously observed at the NH2-terminal end of the membrane-bound penicillinase precursor of Bacillus licheniformis (Yamamoto, S., and Lampen, J . O . (1976) Proc . Natl . Acad . Sci . U . S . A . 73, 1457-1461) . These observations would suggest that bacterial exoproteins which are secreted in the form of precursors differ from extracellular proteins by the presence of an extra segment at their NH2- and/or COOH-terminal ends. Appl Environ Microbiol, 1978 Sep, 36(3), 457 - 64 Effect of phosphate buffer concentration on the heat resistance of Bacillus stearothermophilus spores suspended in parenteral solutions; Gauthier CA et al.; The effect of various quantities of Butterfield phosphate buffer added to four parenteral solutions on the survival of Bacillus stearothermophilus spores heated at 121 degrees C was determined . The effect of the addition of phosphate buffer on spore survival varied with the parenteral solution . Spore survival was increased or decreased, depending upon the composition of the parenteral solution and the buffer concentration . The results obtained in these experiments attest to the fact that environmental factors, including the type of ions present and ionic concentration, affect the heat destruction rate of B . stearothermophilus spores . Therefore, the sterilization requirements of a product such as a parenteral solution may be affected by small changes in formulation. Eur J Biochem, 1978 Sep 1, 89(2), 523 - 9 Biochemical characterization of the restriction-modification system of Bacillus sphaericus; Koncz C et al.; A type II restriction endonuclease (endo R . Bsp) has been purified from Bacillus sphaericus to electrophoretic homogeneity . The enzyme appears to be a single polypeptide chain with a molecular weight of 35000 . Its pH optimum is around 8.2, it requires 20 mM Mg2+ for optimal activity and it is inhibited by Zn2+ . The yield of the enzyme is higher than that of any type II restriction endonuclease so far reported . The enzyme also cleaves single-stranded DNA, albeit at a slower rate . It seems likely that single-stranded DNA is cleaved at the same sequences as double-stranded DNA . Bacillus sphaericus also contains a modification methylase (meth M . Bsp) which completely protects the cell's own DNA against cleavage by its restriction endonuclease . The methylase activity has been partially purified, it copurifies with the nuclease until the next to the last step . The enzyme does not require ATP or Mg2+, it transfers the methyl group of S-adenosyl-methionine to cytosine residues of DNA . As the action of this methylase completely protects any DNA from endo R . Bsp cleavage, it seems likely that the methylase recognizes and methylates the same sequence (dG-dG-dC-dC) as the nuclease. J Virol, 1978 Sep, 27(3), 819 - 22 Sporulation-converting bacteriophages for Bacillus pumilus; Keggins KM et al.; Thirty-three sporulation-converting bacteriophages for Bacillus pumilus NRS576 were assigned to two apparently unrelated groups on the basis of morphology and antiserum neutralization . Bacterial sporulation mutants responded similarly (conversion or nonconversion) to representatives of both phage groups . Evidence is presented indicating that PMB1 and related phages specify a restriction and/or modification system. Carbohydr Res, 1978 Sep, 65(2), 219 - 27 beta-D-xylosidase from Bacillus pumilus PRL B12: hydrolysis of aryl beta-D-xylopyranosides; Kersters-Hilderson H et al.; The influence of substituents on the binding and hydrolysis of several substituted beta-D-xylopyranosides by beta-D-xylosidase from Bacillus pumilus PRL B12 has been investigated . From a comparison of the inhibition constants of 1-thio-beta-D-xylopyranosides with the apparent Michaelis-Menten constants of the substrates, it followed that the latter constants are good approximations of the true equilibrium constants . The influence of the substituent on the rate and activation parameters is small . The results are in agreement with, but do not prove, a one-step mechanism without the formation of a glycosyl-enzyme intermediate. J Bacteriol, 1978 Sep, 135(3), 920 - 7 Oxygen and nitrate in utilization by Bacillus licheniformis of the arginase and arginine deiminase routes of arginine catabolism and other factors affecting their syntheses; Broman K et al.; Bacillus licheniformis has two pathways of arginine catabolism . In well-aerated cultures, the arginase route is present, and levels of catabolic ornithine carbamoyltransferase were low . An arginase pathway-deficient mutant, BL196, failed to grow on arginine as a nitrogen source under these conditions . In anaerobiosis, the wild type contained very low levels of arginase and ornithine transaminase . BL196 grew normally on glucose plus arginine in anaerobiosis and, like the wild type, had appreciable levels of catabolic transferase . Nitrate, like oxygen, repressed ornithine carbamoyltransferase and stimulated arginase synthesis . In aerobic cultures, arginase was repressed by glutamine in the presence of glucose, but not when the carbon-energy source was poor . In anaerobic cultures, ammonia repressed catabolic ornithine carbamoyltransferase, but glutamate and glutamine stimulated its synthesis . A second mutant, derived from BL196, retained the low arginase and ornithine transaminase levels of BL196 but produced high levels of deiminase pathway enzymes in the presence of oxygen. J Bacteriol, 1978 Sep, 135(3), 754 - 9 Effect of growth temperature on membrane fatty acid composition and susceptibility to cold shock of Bacillus amyloliquefaciens; Paton JC et al.; We investigated the fatty acid composition of the membrane of Bacillus amyloliquefaciens grown at different temperatures . A decrease in growth temperature was accompanied by an increase in the ratio of branched- to straight-chain fatty acids and a marked increase in the level of unsaturation of branched-chain fatty acids . When cells of this organism grown at 30 degrees C were cold shocked, viability and ability to secrete extracellular protease were lost . Growth of this organism at lower temperatures or addition of Tween 80 to cells caused the critical temperature zone for cold shocking to be lowered significantly . These results suggest a direct correlation between membrane fluidity and the susceptibility to cold shock. Proc Natl Acad Sci U S A, 1978 Sep, 75(9), 4349 - 53 Modulation of macrophage tumoricidal capability by polyene antibiotics: support for membrane lipid as a regulatory determinant of macrophage function; Chapman HA Jr et al.; We have examined the effects of the sterol-binding polyene antibiotics on macrophage tumoricidal capability . Incubation for 2 hr of activated macrophages from bacillus Calmette-Guerin-infected mice with amphotericin B at 0.5--2 microgram/ml or amphotericin B methyl ester at 0.5--10 microgram/ml enhanced the capability of activated macrophages to kill 3T12 cells . These polyenes did not make normal or stimulated macrophages tumoricidal . Experiments with the ionophores gramicidin, alamethecin, nigericin, and valinomycin indicate that the ionophoretic properties of amphotericin B may not account for its enhancing effect on macrophage tumoricidal potential . Two polyenes with a smaller ring structure, filipin and pimaricin, were also ineffective suggesting that stereospecific modifications in membrane lipid organization underlie the enhancing effect of amphotericin B . The results suggest that the clinical efficacy of amphotericin B in promoting resistance to fungal disease and possibly to neoplasia may operate in part through potentiation of macrophage effector functions. J S Afr Vet Assoc, 1978 Sep, 49(3), 219 - 21 Tuberculosis in laboratory animals; Fourie PB et al.; Tuberculosis can cause great losses in captive colonies of various animal species . In South Africa the culprit is the human type of tubercle bacillus . Interspecific transmission of tuberculosis infection amongst laboratory animals, notably primates, is known to occur, and often handlers and caretakers act as the source of infection . The need for preventive measures in laboratory colonies and procedures for case finding and treatment of tuberculous animals are discussed . Indiscriminate destruction of diseased animals is opposed . The South African situation as revealed by questionnaire survey is described. J Bacteriol, 1978 Sep, 135(3), 841 - 50 Isolation and characterization of Bacillus megaterium mutants containing decreased levels of spore protease; Postemsky CJ et al.; A proteolytic activity present in spores of Bacillus megaterium has previously been implicated in the initiation of hydrolysis of the A, B, and C proteins which are degraded during spore germination . Four mutants of B . megaterium containing 20 to 30% of the normal level of spore proteolytic activity have been isolated . Partial purification of the protease from wild-type spores by a reviewed procedure resulted in the resolution of spore protease activity on the A, B, and C proteins into two peaks--a major one (protease II) and a minor one (protease I) . The protease mutants tested lacked active protease II . All of the mutants exhibited a decreased rate of degradation of the A, B, and C proteins during spore germination at 30 degrees C, but degradation of the proteins did occur . Degradation of the A, B, and C proteins during germination of the mutant spores was decreased neither by blockade of ATP production nor by germination at 44 degrees C . Initiation of spore germination was normal in all four mutants, and all four mutants went through outgrowth, grew, and sporulated normally in rich medium . Similarly, outgrowth of spores of two of the four mutants was normal in minimal medium at 30 degrees C . In the two mutants studied, the kinetics of loss of spore heat resistance and spore UV light resistance during germination were identical to those of wild-type spores . This indicates that the A, B, and C proteins alone are not sufficient to account for the heat or UV light resistance of the dormant spore. Mikrobiologiia, 1978 Sep-Oct, 47(5), 881 - 7 {DNA synthesis and degradation in the cells of Bacillus stearothermophilus}; Trofimenko AF et al.; ATP-dependent and ATP-independent synthesis of DNA was studied on nucleotide-permeable cells of Bacillus stearothermophilus . The effect of temperature, pH, ionic strength, and bivalent metal ions on both types of DNA synthesis was investigated as well as their susceptibility to an SH-blocking agent . The level of ATP-independent synthesis of DNA in the cells of Bac . stearothermophilus was found to be ten times higher than that in E . coli . In the semipermeable cells of Bac . stearothermophilus, 90% of DNA of the chromosome was accessible to their nucleases . The temperature optimum of the activity of DNA polymerases and nucleases in the cells treated with toluene coincided with the optimum of the bacterial growth. J Biol Chem, 1978 Aug 25, 253(16), 5719 - 25 Properties of crystalline leucine dehydrogenase from Bacillus sphaericus; Ohshima T et al.; The distribution of bacterial leucine dehydrogenase (L-leucine:NAD+ oxidoreductase, deaminating, EC 1.4.1.9) was investigated, and Bacillus sphaericus (IFO 3525) was found to have the highest activity of the enzyme . Leucine dehydrogenase, which was purified to homogeneity and crystallized from B . sphaericus, has a molecular weight of about 245,000 and consists of six identical subunits (Mr = 41,000) . The enzyme catalyzes the oxidative deamination of L-leucine, L-valine, L-isoleucine, L-norvaline, L-alpha-aminobutyrate, and L-norleucine, and the reductive amination of their keto analogues . The enzyme requires NAD+ as a cofactor, which cannot be replaced by NADP+ . D-Enantiomers of the substrate amino acids inhibit competitively the oxidation of L-leucine . The enzyme activity is significantly reduced by both sulfhydryl reagents and pyridoxal 5'-phosphate . Purine and pyrimidine bases, nucleosides and nucleotides have no effect on the enzyme activity . Initial velocity and product inhibition studies show that the reductive amination proceeds through a sequential ordered ternary-binary mechanism . NADH binds first to the enzyme followed by alpha-ketoisocaproate and ammonia, and the products are released in the order of L-leucine and NAD+ . The Michaelis constants are as follows: L-leucine (1 mM), NAD+ (0.39 mM), NADH (35 micrometer), alpha-ketoisocaproate (0.31 mM), and ammonia (0.2 M) . The pro-S hydrogen at C-4 of the dihydronicotinamide ring of NADH is exclusively transferred to the substrate; the enzyme is B-stereospecific. Biochemistry, 1978 Aug 22, 17(17), 3468 - 74 Isolation and characterization of polyadenylate-containing RNA from Bacillus brevis; Sarkar N et al.; A substantial fraction (30--40%) of pulse-labeled RNA from exponentially growing cells of Bacillus brevis contains polyadenylate sequences, as measured by adsorption to oligo(dT)-cellulose . The weight-average length of poly(A) tracts obtained after digestion with pancreatic and T1 ribonucleases is 60 nucleotide residues . Susceptibility to degradation by snake venom phosphodiesterase after ribonuclease degradation indicates that the poly(A) sequences are located near the 3' ends of the RNA chains, but that in 40% of the material at least one internal pyrimidine nucleotide residue intervenes between the poly(A) tract and the 3'-hydroxyl terminus . These pyrimidine nucleotides consist of 65% cytidylate and 35% uridylate residues . In the remaining RNA chains, the poly(A) sequence is directly at the 3'-terminus, but the possibility cannot be excluded that a small fraction of this material may contain a 3'-hydroxyl terminal guanylate residue . The weight-average sedimentation coefficient of poly(A)-containing RNA is 12.5 S, corresponding to a polynucleotide chain length of 800--900 residues . This is in a size range expected for messenger RNA, a possibility which is also supported by the observation that pulse-labeled RNA has a considerably higher poly(A) content than long-term labeled RNA. Mol Gen Genet, 1978 Aug 17, 164(2), 195 - 204 Isolation and characterization of plasmid from the Bacillus brevis var . G.-B . cells; Dobritsa AP et al.; The plasmid designated pAD1 was isolated from the cells of four variants of Bacillus brevis var . G.-B . The plasmid DNA has a molecular weight of about 47.1 x 10(6) daltons and contains 43.4 mole % G+C . The bulk of pAD1 DNA (96--98%) is associated with the fraction of chromosome DNA and membranes . Restriction endonucleases Sma I, Sal I and Bam HI cleaved the plasmid DNA into two, two and six fragments, respectively . The cleavage map of the pAD1 genome has been constructed for these three endonucleases . Restriction enzymes Eco RI, Hind III, Kpn I and Pst I hydrolized the plasmid DNA into 16, 21, 10 and 9 fragments, respectively . The presence of repeated sequences in the plasmid genome was shown based on pAD1 DNA cleavage by these endonucleases. Biochem J, 1978 Aug 15, 174(2), 635 - 40 Metabolism and the triggering of germination of Bacillus megaterium . Use of L-{3H}alanine and tritiated water to detect metabolism; Scott IR et al.; L-{2,3-3H}Alanine was used to probe for metabolism of alanine during triggering of germination of spores of Bacillus megaterium KM . No detectable incorporation of label into any compound, including water, was found, indicating that any metabolism involving the alanine germinant must be at a very low rate and also that alanine racemase is absent from spores of this strain . Spores were germinated in 3H2O to find if any of the many metabolic reactions causing irreversible incorporation of 3H into reaction products took place during triggering of germination . No incorporation was detected until 2-3 min after addition of germinants . It is therefore concluded that a wide variety of metabolic routes, including glycolysis, the tricarboxylic acid cycle, the pentose phosphate pathway and amino acid metabolism are either not involved in the reactions causing the triggering of germination or operate at an extremely low rate during this process. Biochem J, 1978 Aug 15, 174(2), 627 - 34 Metabolism and the triggering of germination of Bacillus megaterium . Concentrations of amino acids, organic acids, adenine nucleotides and nicotinamide nucleotides during germination; Scott IR et al.; A considerable amount of evidence suggests that metabolism of germinants or metabolism stimulated by them is involved in triggering bacterial-spore germination . On the assumption that such a metabolic trigger might lead to relatively small biochemical changes in the first few minutes of germination, sensitive analytical techniques were used to detect any changes in spore components during the L-alanine-triggered germination of Bacillus megaterium KM spores . These experiments showed that no changes in spore free amino acids or ATP occurred until 2-3 min after L-alanine addition . Spores contained almost no oxo acids (pyruvate, alpha-oxoglutarate, oxaloacetate), malate or reduced NAD . These compounds were again not detectable until 2-3 min after addition of germinants . It is suggested, therefore, that metabolism associated with these intermediates is not involved in the triggering of germination of this organism. Appl Environ Microbiol, 1978 Aug, 36(2), 392 - 3 D-values of Bacillus pumilus spores on irradiated devices (inoculated product); Prince HN; The D-values of Bacillus pumilus spores on various devices ranged from 0.14 to 0.23 Mrads . The majority of devices displayed D-values equal to or less than the value obtained on filter paper . Increased resistivity was also encountered. Can J Microbiol, 1978 Aug, 24(8), 909 - 14 Growth characteristics of three bacterial isolates from an arctic soil; Nelson LM et al.; Three bacterial isolates, a Pseudomonas sp., a Bacillus sp., and an Arthrobacter sp., commonly isolated from a hummocky sedge-moss meadow at Devon Island, N.W.T., Canada, were selected for further taxonomic characterization and for a study of the effects of temperature and limiting carbon source on growth . Pseudomonas M216 resembled P . putida and Bacillus M153, B . carotarum . Arthrobacter M51 had growth-factor requirements which were more complex than those of any named species of that genus . The temperature ranges of growth indicated that Pseudomonas M216 and Arthrobacter M51 were psychrotrophic while Bacillus M153 was mesophilic . Growth in batch culture at limiting glucose concentrations enabled the calculation of Ks and Y values for each isolate . These were similar to those obtained for other organisms and Pseudomonas M216 and Bacillus M153 showed a high affinity for glucose . The nutritional versatility of Arthrobacter M51 and its ability to grow at low temperatures and the high growth rates and affinity of Pseudomonas M216 for low substrate concentrations may account for their competitive abilities in the natural environment, while the inability of Bacillus M153 to grow at low temperatures may limit its activity in tundra soils. J Bacteriol, 1978 Aug, 135(2), 393 - 401 Membrane phospholipid asymmetry in Bacillus amyloliquefaciens; Paton JC et al.; The phospholipid distribution in the membrane of Bacillus amyloliquefaciens was studied by using phospholipase C (B . cereus), phospholipase A2 (Crotalus), and the nonpenetrating chemical probe trinitrobenzenesulfonic acid . After treatment of intact protoplasts of B . amyloliquefaciens with either phospholipase, about 70% of total membrane phospholipid was hydrolyzed; specifically, about 90, 90, and 30% of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, respectively . Under these conditions, protoplasts remained intact and sealed . However, when protoplasts that were permeabilized by cold-shock treatment were incubated with either of the phospholipases, up to 80% of cardiolipin was hydrolyzed and phosphatidylglycerol and phosphatidylethanolamine were hydrolyzed virtually to completion . In intact cells, 92% of the phosphatidylethanolamine could be labeled with trinitrobenzenesulfonic acid under conditions in which the reagent did not penetrate the membrane to any significant extent . These results indicate that 70% of total phospholipid of this bacillus exists in the outer half of the bilayer . The distribution of phosphatidylethanolamine in this bilayer is highly asymmetric with it being located predominantly in the outer half . The results with phospholipases suggest that the distributions of cardiolipin and phosphatidylglycerol are also asymmetric but independent confirmation of this is required. J Bacteriol, 1978 Aug, 135(2), 363 - 72 Effect of cultural conditions on the concentrations of metabolic intermediates during growth and sporulation of Bacillus licheniformis; Donohue TJ et al.; Intracellular concentrations of adenine nucleotides and intermediates of the Embden-Meyerhof pathway and the tricarboxylic acid cycle have been determined during growth and sporulation of Bacillus licheniformis in a variety of different media . The ATP pool was independent of growth rate and nitrogen source, but the use of glucose as a carbon source resulted in a twofold elevation in the ATP pool during exponential growth . The intracellular phosphoenolpyruvate pool was at least twofold higher during gluconeogenesis than during glycolysis . The finding that the use of glutamate as the sole nitrogen source resulted in at least a fivefold elevation of the alpha-ketoglutarate pool suggests a role for alpha-ketoglutarate in the repression of the enzymes of the tricarboxylic acid cycle responsible for alpha-ketoglutarate synthesis . Not one of the metabolites assayed appears to function as a signal of the nutrient deprivation which accompanies the initiation of sporulation. J Protozool, 1978 Aug, 25(3 Pt 2), 380 - 2 Effect of immunization of cats with Isospora felis and BCG on immunity to reexcretion of Toxoplasma gondii oocysts; Dubey JP; The effect of pretreatment with Isospora felis and bacillus Calmette-Guerin (BCG) on the reexcretion of Toxoplasma gondii oocysts was studied in 16 coccidia-free cats . The following conclusions were drawn: (A) Chronically T . gondii-infected cats reexcreted T . gondii oocysts after superinfection with I . felis, and this reexcretion was prevented in cats infected with I . felis before T . gondii infection . (B) Administration of BCG before Toxoplasma infection had no apparent effect on the outcome of the infection. J Biochem (Tokyo), 1978 Aug, 84(2), 435 - 41 Studies on gramicidin S synthetase . Purification and properties of the light enzyme obtained from some mutants of Bacillus brevis; Kanda M et al.; The phenylalanine-activating and/or-racemizing enzyme, i.e., the light enzyme, of gramicidin S synthetase was purified to a homogenous state by D-phenylalanine-Sepharose 4B chromatography from a wild and some gramicidin S-lacking mutant strains of Bacillus brevis . The light enzyme obtained from a mutant strain E-1 could activate phenylalanine but not racemize it, and had no phenylalanine-dependent ATP-{14C}AMP exchange activity, whereas the same enzyme obtained from other mutants and the wild strain had all three activities . Furthermore, the light enzyme of the mutant E-1 could form only acid-labile enzyme-bound phenylalanine, while the same fraction of the wild strain carried half of the enzyme-bound phenylalanine as acid-labile adenylate and half as a acid-stable thioester . These results suggest that the thiol site of the light enzyme of mutant E-1 might be damaged. J Biochem (Tokyo), 1978 Aug, 84(2), 425 - 34 Studies on gramicidin S synthetase . Purification of the heavy enzyme obtained from some mutants of Bacillus brevis; Hori K et al.; The heavy enzyme of gramicidin S synthetase was purified to an almost homogeneous state by a combination of ammonium sulfate fractionation, ornithine-Sepharose 4B chromatography, DEAE-cellulose chromatography, and Ultrogel AcA 22 chromatography . The enzyme was proved to be essentially homogeneous by ultracentrifugation and polyacrylamide disc gel electrophoresis . The heavy enzymes of gramicidin S synthetase from various groups of mutant strains lacking the ability to form gramicidin S were also purified to a similar extent . The sedimentation rates of the purified enzymes from a wild strain and the mutant strains (BI-3, BII-3, BI-9) were studied by analytical centrifugation and sucrose density gradient centrifugation . The enzymes from the wild strain and these mutant strains were all found to have an S20,W value of 12.2 at a protein concentration of 2.5 mg per ml . These results strongly suggest that the failure of specific amino acid activation in the heavy enzyme of these gramicidin-lacking mutants might be due to some modification at the active center of the corresponding amino acid-activating enzyme rather than to a complete absence of the amino acid-activating enzyme protein in the heavy enzyme. J Biochem (Tokyo), 1978 Aug, 84(2), 467 - 76 Purification and characterization of L-pyrrolidonecarboxylate peptidase from Bacillus amyloiliquefaciens; Tsuru D et al.; Microorganisms capable of producing L-pyrrolidonecarboxylate peptidase {L-pyrrolidonyl peptidase, EC 3.4.11.8} were screened and a strain of Bacillus amyloliquefaciens was chosen as one of the most potent producers of the enzyme . The enzyme was purified from lysozyme-lysate of the bacterial cells by salting out with ammonium sulfate, adsorption on DEAE-cellulose, covalent chromatography on PCMB-Sepharose and by gel filtration on Sephadex G-150 . By these procedures, the enzyme was purified about 800-fold with an activity recovery of 9%, and the preparation was electrophoretically homogenous . The enzyme was most active and stable at pH 7-8 . The presence of 2-mercaptoethanol and EDTA was effective for stabilizing the enzyme . The molecular weight was estimated to be 72,000 by the gel filtration method and to be 24,000 by SDS-polyacrylamide gel electrophoresis, suggesting that the enzyme is a subunit oligomer, presumably trimer . The enzyme was inactivated by the addition of PCMB, sodium tetrathionate, Hg2+ and Cu2+, but the activity lost was restored by the addition of 2-mercaptoethanol and EDTA . The purified enzyme split amide and ester linkages in L-pyroglutamyl derivatives of L-alanine, beta-naphthylamine, alpha-naphthol, and 4-methylumbelliferone, but was completely inert towards various peptides and esters used as substrates for usual amino- and carboxy-peptidases, and for endopeptidases such as trypsin, subtilisin and alpha-chymotrypsin. Eur J Biochem, 1978 Jul 17, 88(1), 135 - 41 Interpretation of the Mössbauer spectra of the four-iron ferredoxin from Bacillus stearothermophilus; Middleton P et al.; The Mossbauer spectra of both oxidized and reduced ferredoxin from Bacillus stearothermophilus have been analysed using computer fits to theoretical spectra obtained from a spin Hamiltonian . A consistent set of parameters was obtained from fits to spectra obtained over a wide range of temperature and magnetic field . These results are interpreted in terms of a model for the active centre which is consistent with its electronic and magnetic properties in both redox states . In the model for the oxidized centre all four iron atoms have essentially the same valence, intermediate between ferric and ferrous, with one pair spin-up and the other pair spin-down . On reduction the extra electron goes predominantly to one pair of iron atoms which become ferrous with the other pair remaining substantially unchanged . Using this model it is possible to obtain relationships between the spin Hamiltonian parameters for individual iron atoms and those for the coupled centre . This can give further insight into the relation between the observed electron paramagnetic resonance and Mossbauer spectra. Eur J Biochem, 1978 Jul 17, 88(1), 275 - 85 Purification and characterization of the penicillin-binding protein that is the lethal target of penicillin in Bacillus megaterium and Bacillus licheniformis . Protein exchange and complex stability; Chase HA et al.; The penicillin-binding protein that is thought to be the lethal target of penicillin in Bacillus megaterium (protein 1) has been purified to greater than 95% homogeneity . The membrane-bound penicillin-binding proteins were solubilized with a non-ionic detergent and partially separated from each other by ion-exchange chromatography on DEAE-Sepharose CL-6B . Protein 1 was subsequently purified by covalent affinity chromatography on ampicillin-affinose . Bacillus licheniformis contains an equivalent penicillin-binding protein (protein 1) that can be more readily purified to virtual homogeneity in a one-step procedure . It was separated from the other penicillin-binding proteins by utilizing the observation that in this organism, this particular protein is the only one whose covalent complex with benzylpenicillin subsequently breaks down . Membranes were treated with saturating concentrations of benzylpenicillin followed by the removal of free penicillin and further incubation to allow the complex between benzylpenicillin and protein 1 to break down . The penicillin-binding proteins were then solubilized and applied to a column of ampicillin-affinose to which only protein 1 was bound as the other penicillin-binding proteins still had benzylpenicillin bound to them . Pure protein 1 was eluted from the affinity resin with hydroxylamine . The interaction of benzylpenicillin with purified protein 1 has been studied by separating unbound antibiotic from the benzylpenicillin . protein complex by paper electrophoresis . Benzylpenicillin reacts with the protein rapidly to form a covalent complex and the fully saturated complex has a molar ratio of bound {14C} benzylpenicillin: protein of 0.7:1 . The complex breaks down, obeying first-order kinetics, with a half-life of 16 min at 35 degrees C, a value identical to that obtained with the membrane-bound protein . The concentration of benzylpenicillin that results in the formation of 50% of the maximum amount of benzylpenicillin . protein complex is that at which the molar amount of benzylpenicillin present is equal to 50% of the molar amount of penicillin-binding protein, rather than being a measure of any of the kinetic parameters of the binding reaction . This observation may be significant in the interpretation of previous results where the amounts of penicillins needed to kill cells or to inhibit penicillin-sensitive reactions have been expressed as concentrations . The possible importance of the breakdown of beta-lactam . protein complexes in the clinical use of these antibiotics is discussed. Biochim Biophys Acta, 1978 Jul 3, 541(3), 301 - 11 An evaluation of respiration chain-associated functions during initiation of germination of Bacillus megaterium spores; Dills SS et al.; In Bacillus megaterium QM B1551, spore germination could be initiated by glucose in the absence of detectable oxygen consumption, ATP synthesis or a pH decrease in the external media, suggesting that none of those reactions were mandatory . In addition, initiation of germination was insensitive to a variety of inhibitors of energy production or protonmotive force uncouplers . Therefore the respiratory chain-associated functions are not prerequisites for initiation of germination but these functions may be necessary to drive energy-dependent transport systems and other biosynthetic reactions during outgrowth. Rev Rhum Mal Osteoartic, 1978 Jul-Sep, 45(7-9), 463 - 8 {Multifocal tuberculous osteoarthritis and synovitis . 10 cases}; David-Chausse J et al.; Synovial or osteo-articular tuberculosis is multifocal in about 10 percent of cases . Here, this form strikes older people and quite frequently the patients receive corticotherapy because of confusion with rheumatism . This premature treatment delays the diagnosis and at the same time favors the spreading of the germ . The 10 patients observed in this study had an average of 3 focuses simultaneously, the sites most often involved being the synovialis of the flexors of the fingers, the discovertebral articulations, the knee and the tibiotarsalis . The diagnosis is based on the presence of fistula (4 times out of 10), x-ray of the lungs (4 miliaries and 1 infiltrate), films of the spine (characteristic images of Pott, 6 times), the bacteriological study (recovery of Koch bacillus 3 times upon direct examination and 6 times after culture) and finally the biopsy of the synovialis (specific synovitis 3 times out of 4 biopsies) . Antituberculosis treatment often involved tolerance problems . It helped to cure 7 patients with few sequelae on the whole . The 3 other patients died either because of old age or complications due to corticosteroids. Prikl Biokhim Mikrobiol, 1978 Jul-Aug, 14(4), 510 - 4 {Biosynthesis of L-asparaginase-2 by cultures of Bacillus polymyxa var . Ross}; Nefelova MV et al.; Cell extracts of Bacillus polymyxa var . Ross.--producer of the polypeptide antibiotic polymyxin M . showed activity of L-asparaginase-2 (L-asparagine aminohydrolase EC 3.5.1.1) . The enzyme activity in the growing culture increased with the biomass . The highest specific activity was detected in the cells at the onset of the stationary stage . The synthesis of L-asparaginase-2 was subjected to glucose catabolite repression in response to its addition to the culture at the logarithmic stage . After purification L-asparaginase-2 was obtained that was 350 times more active than the initial preparation . The enzyme properties were examined. Lab Anim, 1978 Jul, 12(3), 149 - 50 Water-borne Bacillus licheniformis infection in mice; Wright DJ et al.; A water-borne Bacillus licheniformis infection was associated with depressed haemoglobin content, white cell and platelet count . The epidemic was resolved by changing from tanked to mains water supply. J Antibiot (Tokyo), 1978 Jul, 31(7), 652 - 61 The structure of tridecaptin A (studies on antibiotics from the genus Bacillus . XXIV); Kato T et al.; On examining the structure of the antibiotic tridecaptin A, the constituent amino acids were determined to be: 2,4-diaminobutyric acid(2D, 1L), Ser(1D, 1L), Glu(1L), Gly(1), Ala(1L), Val(1D, 1L), aIle(1D), Phe(1L) and Trp(1D) . The constituent fatty acid was identified as beta-hydroxy anteisononanoic acid by gas chromatography and mass spectrometry . Cleavage reaction with N-bromosuccinimide, sequential analysis by EDMAN degradation, partial acid hydrolysis and some additional evidences clarified the structure of tridecaptin A. J Antibiot (Tokyo), 1978 Jul, 31(7), 646 - 51 Isolation of tridecaptins A, B and C (studies on antibiotics from the genus Bacillus . XXIII); Shoji J et al.; Three new antibiotics, tridecaptins A, B and C, were isolated from culture broths of strains of Bacillus polymyxa AR-110, B-2 and E-23, respectively . All are acyl tridecapeptides differing from each other in the fatty acid components and amino acid residues . They are weakly active against Gram-negative and Gram-positive bacteria in vitro and in vivo. J Clin Microbiol, 1978 Jul, 8(1), 108 - 9 Group IVe-like gram-negative bacillemia in a patient with obstructive uropathy; Rockhill RC et al.; A nonfermentative, gram-negative bacillus, resembling Center for Disease Control group IVe, was isolated from the blood of a patient with obstructive uropathy . Prior studies implicating this organism in septicemia could not be found in the literature . It is suggested that this is the first time that this bacillus has been reported to cause bacillemia in humans. Cancer Treat Rep, 1978 Jul, 62(7), 1085 - 7 Chemoimmunotherapy for disseminated malignant melanoma: a prospective randomized study; Ramseur WL et al.; Twenty-eight patients with disseminated malignant melanoma were treated with DTIC (250 mg/m2 iv, Days 1--5) and actinomycin D (0.5 mg/day iv, Days 1--5) at 5-week intervals . Patients were randomly allocated to receive no immunotherapy or immunotherapy consisting of methanol extracted residue of bacillus Calmette-Guerin (0.5 mg intradermally; 100 microgram in five separate sites) every 5 weeks, concomitant with chemotherapy . Of these 28 evaluable patients, 13 received chemoimmunotherapy with one complete response (CR) and 15 received chemotherapy alone with one CR . Both responses occurred in lymph node metastases . No partial responses were seen. Nucleic Acids Res, 1978 Jul, 5(7), 2267 - 88 Binding sites of E . coli and B . stearothermophilus ribosomal proteins on B stearothermophilus 5S RNA; Zimmermann J et al.; The primary binding sites for Bacillus stearothermophilus proteins B-L5 and B-L22 and the Escherichia coli proteins E-L5, E-L18 and E-L25 on B . stearothermophilus 5S RNA were determined by limited ribonuclease digestion of the corresponding 5S RNA-protein complexes . The results obtained in this study are in agreement with our previous experiments in which the binding sites of E . coli and B . stearothermophilus proteins were determined for E . coli 5S RNA and lead to the conclusion that the proteins interact with the most conserved regions of 5S RNA . A comparison of the results obtained in this study with those of other published experiments suggest that the proposed interaction of nucleotides 16-21 with those of 58-63 is facilitated by protein binding to 5S RNA. J Exp Med, 1978 Jul 1, 148(1), 288 - 300 Trypanosoma cruzi: in vitro induction of macrophage microbicidal activity; Nogueira N et al.; Normal, resident and inflammatory mouse peritoneal macrophages can be induced to display microbicidal activity against trypomastigotes of Trypanosoma cruzi by exposure to products from antigen-pulsed, sensitized spleen cell populations . Optimal macrophage microbicidal activity was achieved by constant exposure and daily renewal of the spleen cell factors . Macrophages obtained after an intraperitoneal injection of mild inflammatory agents were rapidly induced, displaying trypanocidal activity 24 h after exposure to the active spleen cell factor(s), and by 48 h, parasites were no longer observed . Resident peritoneal macrophages required 24 h longer for activation . Removal of the factor(s) before achieving complete disappearance of intracellular parasites led to resumed growth of the surviving organisms . The spleen cell factor(s) is effective when added either before or after exposure of the macrophages to trypomastigotes, and does not itself alter parasite viability . Dilution of the factor(s) up to 1:16 still results in significant trypanocidal activity . In vivo activated cells, obtained after a specific secondary challenge of animals infected with T . cruzi or Bacille Calmette-Guerin, lose their trypanocidal activity under in vitro conditions . This loss of activity can be prevented or restored by the addition of the active spleen cell factor(s) . Induction of trypanocidal activity is also obtained with products from Concanavalin A- or lipopolysaccharide-stimulated normal spleen cells. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1978 Jul-Sep, 27(3), 161 - 8 {Clinical studies of the role of the fluorescent circulating antibody test in the diagnosis of active tuberculosis}; Albu I et al.; The value was studied, of the test of circulating fluorescent antibodies against M . tuberculosis in establishing the diagnosis of active tuberculosis in 610 patients classified in 4 groups: 1 . active tuberculosis bacteriologically confirmed; 2 . active tuberculosis with negative bacteriological examination, but confirmed by other methods of criteria; 3 . stabilized tuberculosis; 4 . non-tuberculous affections . The specificity of the test in active tuberculosis was of 80% in adults and of 91,5% in children . The test was negative in 80,3% of stabilized tuberculosis cases and in 69% of non-tuberculous affections . The sensitivity was lower in active tuberculosis with positive bacilloscopy (67%) against active tuberculosis with negative bacilloscopy (90,6%), a fact explained by a high consumption of anti-bacillary antibodies, demonstrated by positivation of the test when bacilloscopy becomes negative . The use of the test is indicated in establishing a therapeutic test, and in the context of clinical and radiologic explorations, as well as in the expertising of working capacities of previous tuberculosis patients. Biochem J, 1978 Jul 1, 173(1), 45 - 52 Purification and properties of glutamate synthase and glutamate dehydrogenase from Bacillus megaterium; Hemmila IA et al.; Bacillus megaterium N.C.T.C . no . 10342 exhibits glutamate synthetase (EC 2.6.1.53) and glutamate dehydrogenase (EC 1.4.1.4) activities . Concentrations of glutamate synthase were high when the bacteria were grown on 3mM-NH4Cl and low when they were grown on 100mM-NH4Cl, whereas glutamate dehydrogenase concentrations were higher when the bacteria were grown on 100mM-NH4Cl than on 3mM-NH4Cl . Glutamate synthase and glutamate dehydrogenase were purified to homogeneity from B . megaterium grown in 10mM-glucose/10mM-NH4Cl . The purified enzymes had mol.wts . 840000 and 270000 for glutamate synthase and glutamate dehydrogenase respectively . The Km values for substrates with NADPH and coenzyme were (glutamate synthase activity shown first) 9 micron and 360 micron for 2-oxoglutarate, 7.1 micron and 8.7 micron for NADPH, and 0.2 mM for glutamine and 22 mM for NH4Cl, similar values to those of enzymes from Escherichia coli . Glutamate synthase contained NH3-dependent activity (different from authentic glutamate dehydrogenase), which was enhanced 4-fold during treatment at pH 4.6 NH3-dependent activity was generally about 2% of the glutamine-dependent activity . Amidination of glutamate synthase by the bi-functional cross-linking reagent dimethyl suberimidate inactivated glutamine-dependent glutamate synthase activity, but increased NH3-dependent activity . A cross-linked structure of mol.wt . approx 200000 was the main product formed. Tumori, 1978 Jul-Aug, 64(4), 353 - 60 Bacillus of Calmette and Guérin and 7, 12-dimethylbenz(a)-anthracene carcinogenesis in mice; Scelsi R et al.; The effects of single or repeated Bacillus of Calmette and Guerin (BCG) treatment on the onset of tumors induced by the carcinogen 7,12-dimethylbenz(a)anthracene (DMBA) injected at birth into Swiss mice were studied . Multiple doses of BCG, given at regular intervals significantly lowered the incidence of carcinogen-induced tumors . The DMBA-induced lung adenomas did not differ morphologically from the ones that appeared in mice simultaneously treated with both DMBA and BCG. Can J Microbiol, 1978 Jul, 24(7), 818 - 26 {Study of lysogeny in Bacillus thuringiensis and B . cereus}; Ackermann HW et al.; Forty-eight strains of Bacillus thuringiensis and 12 strains of B . cereus were treated with ultraviolet light and mitomycin C . The former agent was the more effective inducer . Bacillus thuringiensis produces at least seven different phage particles with long, non-contractile tails . The frequencies of lysogeny and polylysogeny are 83 and 25% respectively . Morphologically defective phages occur in 25% of strains, whereas five of them produce low molecular-weight bacteriocins . One strain of B . cereus harbors "killer-particles." There is no apparent correlation between the presence of phage-like particles, phage senstivity, and serotypes, biotypes, or the origin of B . thuringiensis strains. Can J Microbiol, 1978 Jul, 24(7), 798 - 803 Microbial degradation of {C14C}polystyrene and 1,3-diphenylbutane; Sielicki M et al.; Microbial degradation of {beta-14C}polystyrene and 1,3-diphenylbutane, a compound structurally representing the smallest repeating unit of styrene (dimer), was investigated in soil and liquid enrichment cultures . Degradation rates in soil, as determined by 14CO2 evolution from applied {14C}polystyrene, varied from 1.5 to 3.0% for a 4-month period . Although relatively low, these percentages were 15 to 30 times greater than values previously reported . Enrichment cultures, containing 1,3-diphenylbutane as the only carbon souce, were used to determine the mechanisms of microbial oxidation of the polymer chain ends . Metabolism of 1,3-diphenylbutane appeared to involve the attack by a monooxygenease to form 2-phenyl-4-hydroxyphenylbutane followed by a further oxidation and subsequent fission of the benzene ring to yield 4-phenylvaleric acid and an unidentified 5-carbon fragment via the classic meta-fission pathway . Phenylacetic acid was probably formed from 4-phenylvaleric acid by subsequent beta-oxidation of the side chain, methyl-oxidation and decarboxylation . An initial examination of the population of microorganisms in the diphenylbutane enrichment cultures indicated that these oxidative reactions are carried out by common soil microorganism of the genera Bacillus, Pseudomonas, Micrococcus, and Nocardia. J Bacteriol, 1978 Jul, 135(1), 68 - 70 Polyethylene glycol-induced fusion of heat-inactivated and living protoplasts of Bacillus megaterium; Fodor K et al.; Protoplasts of Bacillus megaterium, incubated at 50 degrees C for 120 min, lost the ability to revert to bacillary form . Such heat-inactivated protoplasts, however, produced recombinants when fused by polyethylene glycol treatment with normal protoplasts . Although this differential inactivation effect is not yet fully reproducible, reciprocal inactivations of the parental protoplasts in genetic crosses have clearly shown that for protoplast fusion (i) either of the parents may serve as the viable recipient for markers coming from the heated parental protoplasts, and (ii) either of the parents may be rendered nonviable and yet, when fused with a viable partner, contribute to formation of a recombinant . Heat inactivation seems to provide a way to counterselect when few markers are available and one of the parents is prototrophic. J Bacteriol, 1978 Jul, 135(1), 133 - 7 Study of calcium dipicolinate release during bacterial spore germination by using a new, sensitive assay for dipicolinate; Scott IR et al.; The release of calcium and dipicolinic acid from spores of Bacillus megaterium KM during L-alanine-induced triggering of germination has been studied using a new, simple, and rapid assay for dipicolinic acid capable of detecting a concentration of 0.5 micron . The release of both calcium and dipicolinate started within seconds of exposure of the spores to L-alanine, thus preceding other measurable changes associated with germination . From the earliest times, the two substances were released in equimolar quantities, although later in germination calcium predominated. Digestion, 1978 Jul-Aug, 17(4), 332 - 45 Bacillus cereus-induced malabsorption in young mice; Madge DS; Following a single, oral dose of Bacillus cereus (2 X 10(8) bacteria) in vitro intestinal absorption of D-glucose, D-galactose, L-arginine, L-histidine, L-ornithine and L-proline in young mice (aged 2--3 1/2 months) decreased . Malabsorption of D-glucose was dose- and time-dependent . Impaired absorption of D-glucose occurred throughtout the length of the small intestine, particularly distally . Following hydrolysis of D-maltose at the brush border, D-glucose absorption in infected mice and that of the untreated controls was similar . Using D-glucose, fluid transfer in the infected intestine and that of the controls was alike . Although slightly lower, fluid transfer in the infected intestine using the other solutes was not significantly different compared with the controls . Glucose-dependent and glucose-independent intestinal fluid transfer in infected animals was like that of the controls . Using old infected mice (aged 8--9 months) intestinal absorption of D-glucose and L-histidine was unchanged compared with young mice . The fresh small intestinal weight in infected mice and the controls was alike . Changes in the histology of the small intestine in young infected mice were small and inconsistent. Ann Intern Med, 1978 Jul, 89(1), 64 - 6 Lymph-node bacilliform bodies resembling those of Whipple's disease in a patient without intestinal involvement; Mansbach CM 2nd et al.; A 38-year-old man developed symptoms of arthalgias and arthritis, lymphadenopathy, and weight loss . An axillary lymph-node biopsy was done in the diagnostic study; a periodic acid Schiff stain, done for evidence of fungal infection, showed periodic acid Schiff reagent-positive macrophages . Electron microscopy showed the typical morphologic features of the bacilliform bodies associated with Whipple's disease to be present in the macrophages of the lymph node . The patient had no intestinal symptoms . The absorption of a variety of substrates was found to be normal . Nine intestinal biopsies showed no organisms similar to those found in his lymph node . On tetracycline therapy, he symptomatically improved . The findings raise the question of the route of infection in Whipple's disease and point up the usefulness of periodic acid Schiff staining of lymph-node biopsies. Biochem J, 1978 Jul 1, 173(1), 53 - 8 Inactivation of glutamate dehydrogenase and glutamate synthase from Bacillus megaterium by phenylglyoxal, butane-2,3-dione and pyridoxal 5'-phosphate; Hemmila IA et al.; Reaction of phenylglyoxal with glutamate dehydrogenase (EC 1.4.1.4), but not with glutamate synthase (EC 2.6.1.53), from Bacillus megaterium resulted in complete loss of enzyme activity . NADPH alone or together with 2-oxoglutarate provided substantial protection from inactivation by phenylglyoxal . Some 2mol of {14C}Phenylglyoxal was incorporated/mol of subunit of glutamate dehydrogenase . Addition of 1mM-NADPH decreased incorporation by 0.7mol . The Ki for phenylglyoxal was 6.7mM and Ks for competition with NADPH was 0.5mM . Complete inactivation of glutamate dehydrogenase by butane-2,3-dione was estimated by extrapolation to result from the loss of 3 of the 19 arginine residues/subunit . NADPH, but not NADH, provided almost complete protection against inactivation . Butane-2,3-dione had only a slight inactivating effect on glutamate synthase . The data suggest that an essential arginine residue may be involved in the binding of NADPH to glutamate dehydrogenase . The enzymes were inactivated by pyridoxal 5'-phosphate and this inactivation increased 3--4-fold in the borate buffer . NADPH completely prevented inactivation by pyridoxal 5'-phosphate. Experientia, 1978 Jun 15, 34(6), 762 - 3 Bacillus thuringiensis delta-endotoxin: evidence that toxin acts at the surface of susceptible cells; Fast PG et al.; Enzymically activated delta-endotoxin of Bacillus thuringiensis covalently bound to Sephadex beads, has the same effect on insect cells in tissue culture as free toxin . The effect is prevented by antitoxin antibody and heat denaturation and is not due to a nonspecific protein effect, the beads, or toxin released from the beads . The toxin, therefore, probably acts at the cell surface. J Antibiot (Tokyo), 1978 Jun, 31(6), 525 - 32 Aurantinin, a new antibiotic of bacterial origin; Nishikiori T et al.; A new polyene class antibiotic, aurantinin (KM-214), was isolated from the fermentation broth of Bacillus aurantinus MASUMA and OMURA sp . nov . The substance is a conjugated triene with a molecular weight of 618 and molecular formula C35H54O9, and melts at 139 approximately 140 degrees C . The antibiotic is active in vitro against Gram-positive bacteria, but not against yeast and fungi. Antibiotiki, 1978 Jun, 23(6), 506 - 8 {Antibacterial activity and bacteriophages of Bacillus pumilus}; Abramova MA et al.; Inducable defective phages analogous to those in Bac . subtilis were found in 5 strains of Bac . pumilus, 3 of which possessed antibacterial activity . A moderate bacteriophage was also induced in addition to the defective phage in strain ATCC 12140 not producing the antibiotic . It was found that the antibacterial activity was not associated with the presence of phages in the cultures . The data on the electron microscopic investigations are presented. Am J Clin Pathol, 1978 Jun, 69(6), 637 - 41 Malacoplakia of the endometrium, a probable cause of postmenopausal bleeding; Thomas W Jr et al.; A 60-year-old woman, 20 years post-menopausal, who had deforming rheumatoid arthritis of 7 years' duration and Sjogren's syndrome of 1 year's duration, had had postmenopausal bleeding for a month prior to admission to the hospital . A diagnosis dilatation and curettage was interpreted as showing acute suppurative endometritis . The patient was discharged, only to have recurrent vaginal bleeding . She was readmitted five weeks later, at which time results of another dilatation and curettage were interpreted as showing xanthromatous endometritis . Total hysterectomy with bilateral salpingo-oophorectomy was done . Examination of Epon-embedded endometrium 1 micrometer thick by light microscopy and subsequently by electron microscopy disclosed intracellular bacilliform organisms within phagolysosomes of atypical histiocytes, lamellar bodies, and various developing stages of calcospherites, Michaelis-Gutmann bodies . The curettings were then received and classic Michaelis-Gutmann bodies were identified in periodic acid--Schiff-stained sections. J Bacteriol, 1978 Jun, 134(3), 699 - 705 Relationship between the heat resistance of spores and the optimum and maximum growth temperatures of Bacillus species; Warth AD; Heat resistance of spores of Bacillus strains was compared with the temperature adaptation of each strain as measured by the optimum and maximum growth temperatures and the heat resistance of vegetative cells . Maximum growth temperatures ranged from 31 to 76 degrees C and were little affected by the nature of the growth medium . The temperature giving maximum growth rate was closely correlated to the maximum temperature for growth, and about 6 degrees C lower . Vetetative-cell heat resistance, determined on exponential-phase cells, was also correlated with maximum growth temperature . The temperature at which spores were inactivated with a decimal reduction time of 10 min was in the range of 75 to 121 degrees C . This temperature was 46 +/- 7 degrees C higher than the maximum growth temperature and correlated with it and the other cell parameters . Spore heat resistance can be considered to have two components, the temperature adaptation characteristic of the species and the stabilization conferred by the spore state. Cancer, 1978 Jun, 41(6), 2456 - 63 Intralesional treatment of recurrent metastatic cutaneous malignant melanoma: a randomized prospective study of intralesional Bacillus Calmette-Guerin versus intralesional dinitrochlorobenzene; Cohen MH et al.; Eighteen patients with multiple recurrences of malignant melanoma without evident distant spread were randomly assigned to treatment with either intralesional Bacillus Calmette-Guerin (BCG) or intralesional dinitrochlorobenzene (DNCB) . Both agents were able to destroy approximately 90% of the injected intradermal nodules . Intradermal disease was more easily obliterated than subcutaneous disease with intralesional treatment with either agent, and local control of satellitosis with elimination of all clinically evident tumor was achieved in the patients who had intradermal without subcutaneous satellitosis, regardless of whether the patient was receiving BCG or DNCB . The clinical courses of the treated patients were essentially the same . Although PHA reactivity was depressed, the patients in both groups were responsive to recall and melanoma skin test antigens, demonstrated leukocyte migration inhibition with melanoma antigen and were generally within normal limits when assayed for 29 degrees C E rosettes . Our study demonstrated a dramatic difference in toxicity between the two intralesional agents without a similar difference in therapeutic efficacy or immune testing. Cancer Res, 1978 Jun, 38(6), 1626 - 32 Generation of anti-MOPC-315 cytotoxicity in uneducated or in vitro educated spleen cells from normal or MOPC-315 tumor-bearing mice pretreated in vivo with Bacillus Calmette-Guérin; Braun DP et al.; Cultured spleen cells from normal or MOPC-315 tumor-bearing BALB/c mice that were pretreated in vivo with Bacillus Calmette-Guerin (BCG) exhibited in vitro cytotoxicity against MOPC-315 plasmacytoma . In vitro education of BALB/c spleen cells from normal or tumor-bearing mice by cocultivation with mitomycin C-treated MOPC-315 stimulator cells also resulted in antitumor cytotoxicity . The combination of BCG pretreatment of donor mice with the in vitro education of their spleen cells resulted in a level of anti-MOPC-315 cytotoxicity that was greater than the sum of the levels of cytotoxicity exhibited by spleen cells subjected to either process alone . The levels of cytotoxicity exhibited by educated or uneducated spleen cells from BCG-pretreated mice were dependent on the dose of BCG used and on the time interval between in vivo pretreatment and the initiation of in vitro culture . Thus, our findings suggest that educated spleen cells from tumor-bearing hosts that were pretreated with BCG might be useful in immunotherapeutic regimens requiring histocompatible cells with augmented antitumor cytotoxicity. Surgery, 1978 Jun, 83(6), 677 - 81 Randomized trial of adjuvant therapy for "high risk" primary malignant melanoma; Wood WC et al.; Retrospective pathological classification of 213 patients with malignant melanoma identified a group at high risk of recurrence (25% developed recurrence in 12 months, 50% by 5 years) after resection for apparent cure . Using these criteria, 70 patients were identified after resection of all apparent disease as being at high risk for recurrent melanoma . They were randomly assigned to one of the three adjuvant treatment arms: chemotherapy with dimethyl triazeno imidazole carboxamide (DTIC), immunotherapy with bacillus Calmette-Guerin (BCG), or combined chemoimmunotherapy . Six of 20 patients receiving DTIC developed recurrence (30%) and four died (20%) . Five of 28 patients receiving BCG developed recurrence (18%) and two died (7.5%) . There have been no recurrences or deaths in 22 patients receiving combined chemoimmunotherapy . In the prevention of early recurrence, the combined therapy arm was significantly superior to both the immunotherapy arm (p less than 0.05) and the chemotherapy arm (p less than 0.01) . In terms of survival, combined therapy also was superior to chemotherapy alone (p less than 0.05). J Hyg (Lond), 1978 Jun, 80(3), 321 - 5 The sensitization of children by opportunist mycobacteria in Lagos, Nigeria; Ogunmekan DA; Groups of school children aged 6-14 years were tested with PPDS and one of the following five antigens simultaneously, namely PPDA, PPDF, PPDG, PPDPL and PPDY, PPDS was prepared from the human tubercle bacillus, and the others were prepared from M . avium, M . fortuitum, the 'Gause' organisms, M . marinum and M . kansasii respectively . It was observed that there was some increase in induration size to PPDA, PPDG, PPDL, PPDS and PPDY with increase in age, while M . fortuitum gave a preponderance of small reactions with no increase of size with age . It was also observed that there was evidence of cross sensitization between PPDS and all the other antigens, particularly in those who had negative or doubtful reactions. Hautarzt, 1978 Jun, 29(6), 331 - 6 {Psoriasis and leprosy in the light of common history . A contribution on epidemiology and differential diagnosis}; Eckes LK; Over the centuries psoriasis was described as a variety of leprosy . It is only in the last century, however, that it was described as a separate entity . Many early medical descriptions of this woridwide affliction can be found, which attempt to distinguish between the various kinds of leprosy . These attempts led to cultural-religious consequences on one hand and to different theories about their contagiousness and curability on the other . Today the current conviction, that with the proof for the existence of a lepra bacillus the question of heritability of the disease can finally be discarded, is being reconsidered . For both psoriasis and leprosy the search for relationships between these diseases and genetically determined markers in the blood-, serum protein- and enzyme-group systems has led to applicable results which, insofar as they can be interpreted as indications of selective factors, will enable us to understand the geographical distribution of both diseases. P N G Med J, 1978 Jun, 21(2), 158 - 61 Sepik granuloma; Aiken GH et al.; An unusual infection occuring in the East Sepik Province of Papua New Guinea is reported . The patients come from villages on the Sepik River or its tributaries . The lesions consist of cutaneous nodules and papillomas which are slowly progressive . An unidentified organism, apparently a gram positive bacillus, is seen in large numbers in the lesions; a natural habitat in soil or water or on vegetation seems likely. Inflammation, 1978 Jun, 3(2), 159 - 76 The effect of cortisone on the accumulation, activation, and necrosis of macrophages in tuberculous lesions; McCue RE et al.; Rabbits were injected intramuscularly with cortisone acetate (2 mg/kg) on alternate days . Six days after the first injection these rabbits and controls were injected intradermally in multiple sites with BCG (the vaccine strain of tubercle bacillus) . Periodically, over the next 2 months, the resulting lesions were measured and surgically biopsied, and the animals were tuberculin-tested . Macrophage activation in the BCG lesions was evaluated histochemically by staining for beta-galactosidase activity . Both BCG lesions (and tuberculin reactions) in the cortisone-treated group were considerably smaller than those in the control group . Cortisone was highly effective in reducing the number of infiltrating mononuclear cells (MN), the amount of caseous necrosis and ulceration, and the percent of NM that were beta-galactosidase-positive . The decreased activation and reduced number of macrophages readily explains the increased susceptibility to tuberculosis found amoung patients receiving glucocorticosteroids . In the BCG lesions, the local decrease in the number and function of leukocytes probably explains the decreased tissue necrosis . Such antiinflammatory effects of corticosteroids may offset, in selected antimicrobial-treated cases, the hormone's detrimental effect on host resistance to infectious agents. Br J Cancer Suppl, 1978 Jun, 37(3), 34 - 7 Effects of some hypoxic cell radiosensitizers on the decay of potentially lethal oxygen-dependent damage in fully hydrated spores; Tallentire A et al.; Using a stopped-flow mixing and pulsed irradiation apparatus, a study has been made of the decay, to a harmless form, of radiation-induced species that would otherwise be lethal to spores on contact with oxygen . Aqueous suspensions of Bacillus megaterium spores were irradiated with electrons for approximately 1 s; at various times after irradiation oxygen in solution was added . As the interval between anoxic irradiation and introduction of oxygen increased, the fraction of spores surviving increased . This change in survival reflects the decay of potentially lethal species . The presence of electron-affinic radiosensitizers during irradiation enhanced the decay rate of this damage, the greatest enhancement being seen with sensitizers of the highest electron affinity . In contrast, the nitroxyl-free radical sensitizer TAN fixed the radiation-induced damage so that no increase in survival, and hence no decay, was seen. C R Acad Sci Hebd Seances Acad Sci D, 1978 Jun, 286(22), 1629 - 32 {Cytological study of the action of Bacillus thuringiensis var . israelensis on mosquito larvae}; Barjac H; A comparison is made between the cytopathological effects of B . thuringiensis var . israelensis on Aedes aegypti larvae and the cytopathological effects of the other varieties of B . thuringiensis on Lepidoptera larvae . The same primary action is observed, with the loss of integrity of the gut epithelium, the cells of which appear swollen, distorted and finally burst. J Bacteriol, 1978 Jun, 134(3), 1157 - 70 Properties of Bacillus cereus temperature-sensitive mutants altered in spore coat formation; Stelma GN Jr et al.; Three conditional Bacillus cereus mutants altered in the assembly or formation of spore coat layers were analyzed . They all grew as well as the wild type in an enriched or minimal medium but produced lysozyme and octanol-sensitive spores at the nonpermissive temperature (35 to 38 degrees C) . The spores also germinated slowly when produced at 35 degrees C . Temperature-shift experiments indicated that the defective protein or regulatory signal is expressed at the time of formation of the outer spore coat layers . Revertants regained all wild-type spore properties at frequencies consistent with initial point mutations . Spore coat defects were evident in thin sections and freeze-etch micrographs of mutant spores produced at 35 degrees C . In addition, one mutant contained an extra surface deposit, perhaps unprocessed spore coat precursor protein . A prevalent band of about 65,000 daltons (the same size as the presumptive precursor) was present in spore coat extracts of this mutant and may be incorrectly processed to mature spore coat polypeptides . Another class of mutants was defective in the late uptake of half-cystine residues into spore coats . Such a defect could lead to improper formation of the outer spore coat layers. J Bacteriol, 1978 Jun, 134(3), 1081 - 8 Hybridization analysis of restriction endonuclease DNA fragments of Bacillus cereus transcribed during spore outgrowth; Silberstein Z et al.; Transcribing Bacillus cereus DNA was visualized by means of autoradiography of electrophoretically separated EcoRI restriction endonuclease DNA fragments hybridizing 32P-labeled RNA . Hybridization of RNA of dormant spores, vegetative cells, and outgrowing spores indicates the following . (i) A large fraction of the nonribosomal RNA in dormant spores is transcribed at a limited number of regions on the bacterial chromosome . (ii) After induction of spore germination, transcription activity is not limited to a single short region on the chromosome, but rather is distributed along the chromosome . The DNA/RNA hybridization technique has been used to identify restriction endonuclease DNA fragments homologous to RNA species that are present in dormant spores but absent from vegetative cells, RNA species that are synthesized immediately after germination induction and are present at a relatively low concentration in vegetative cells, and RNA species that are transcribed at a late stage of outgrowth but are absent or present at low concentration at an early stage of outgrowth. Appl Environ Microbiol, 1978 Jun, 35(6), 1155 - 9 Simple method for the isolation of astaxanthin from the basidiomycetous yeast Phaffia rhodozyma; Johnson EA et al.; A method is described for the quantitative and, possibly, large-scale extraction of astaxanthin from the yeast Phaffia rhodozyma . The method utilizes extracellular enzymes produced by the bacterium Bacillus circulans WL-12, which partially digests the yeast cell wall and renders the carotenoid pigments extractable by acetone or ethanol . Complete recovery of astaxanthin from heat-killed P . rhodozyma cells was obtained after growing B . circulans WL-12 on these yeast cells for 26 h and then extracting the yeast-bacterium mixture with acetone . A bacteria-free lytic system, which gave quantitative extraction of astaxanthin from P . rhodozyma, was obtained by concentrating the culture broth from the growth of B . circulans WL-12 on P . rhodozyma cells . Hydrolytic enzyme activities detected in this concentrate included beta-(1 leads to 3)-glucanase, beta-(1 leads to 6)-glucanase, alpha-(1 leads to 3)-glucanase, xylanase, and chitinase . The lytic system was found to work most efficiently at pH 6.5 and with low concentrations of yeast. C R Acad Sci Hebd Seances Acad Sci D, 1978 May 29, 286(21), 1535 - 8 {Modes of tetracycline resistance in Bacillus thuringiensis}; Fargette F et al.; The sensitive strain and tetracycline-resistant mutants exhibit two types of growth curves after addition of tetracycline . The level of resistance of these strains is enhanced by a short period of preincubation with a non-inhibitive concentration of tetracycline. J Biol Chem, 1978 May 25, 253(10), 3660 - 5 Stability of D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid in relation to its possible occurrence as a degradation product of penicillin by the exocellular DD-carboxypeptidase-transpeptidase from Streptomyces R61 and the membrane-bound dd-carboxypeptidase from Bacillus stearothermophilus; Adriaens P et al.; The stability of D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid has been studied under various conditions . In 10 mM cacodylate, pH 6.5, and at 55 degrees C, D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid (at concentrations lower than 1 mM) is hydrolyzed into N-formyl-D-penicillamine with a half-life of 3 to 4 min . On this basis, it is very unlikely that D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid could be one of the end products resulting from the cleavage of benzylpenicillin by the DD-carboxypeptidase of Bacillus stearothermophilus (as reported by Hammarstrom and Strominger (1976) J . Biol . Chem . 251, 7947--7949) . In 3 mM phosphate, pH 7.5, and at 37 degrees C, D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid (at concentrations lower than 1 mM) has a half-life of 45 min . On the basis of kinetic experiments carried out under these conditions with phenoxymethylpenicillin and the DD-carboxypeptidase-transpeptidase of Streptomyces R61, it is concluded that the primary product which arises from the thiazolidine moiety of the antibiotic molecule and gives rise to N-formyl-D-penicillamine, has a half-life of 10 min, a value which is not compatible with the hypothesis that D-5,5-dimethyl-delta2-thiazoline-4-carboxylic acid would be an intermediate involved in the fragmentation pathway. J Biol Chem, 1978 May 25, 253(10), 3595 - 601 Enzymatic formation of uridine diphosphate N-acetyl-D-mannosamine; Kawamura T et al.; An enzyme that catalyzes the interconversion of UDP-N-acetyl-D-glucosamine and UDP-N-acetyl-D-mannosamine was purified about 700-fold from the supernatant fraction of Bacillus cereus, and the properties of this enzyme were studied . This enzyme was not stimulated by NAD+, NADH, or any metal ions . The optimum pH was between 7.5 and 8.0 . At equilibrium of the reaction, the ratio of UDP-N-acetylglucosamine to UDP-N-acetylmannosmaine was about 9:1 . The enzyme was inactive toward free N-acetylhexosamines, their phosphate esters, UDP-glucose, and UDP-N-acetylgalactosamine . A stimulatory role of UDP-N-acetylglucosamine was demonstrated . In the reaction with UDP-N-acetylglucosamine, the rate as a function of substrate concentration showed a sigmoidal relationship with a Hill coefficient of 1.8 and an apparent Km value for UDP-N-acetylglucosamine of 1.1 mM . The reverse reaction with UDP-N-acetylmannosamine required the presence of UDP-N-acetylglucosamine . The UDP-N-acetylglucosamine concentration required for half-maximal activation was about 0.5 mM . The apparent Km for UDP-N-acetylmannosamine measured in the presence of 0.5 mM UDP-N-acetylglucosamine was 0.22mM . Other nucleotides or hexosamine derivatives were not stimulatory . The same activity was found in cell extracts from several bacterial species. C R Acad Sci Hebd Seances Acad Sci D, 1978 May 16, 286(19), 1403 - 5 {Isolation of bacteria that use that use nitric oxide as a respiratory electron acceptor under anaerobiosis}; Pichinoty F et al.; Ten bacteria of the genus Bacillus were isolated from pasteurized soils, in anaerobiosis and at 32 degrees C, on peptone broth containing 0.5% KNO2 . They are Gram variable rods producing oval spores . They are oxidase positive and have catalase . They grow, in anaerobiosis, on NO-3, NO-2, N2O, and NO as respiratory electron acceptors . These compounds are reduced to N2. Biochim Biophys Acta, 1978 May 4, 509(1), 129 - 35 Biochemical aspects of the visual process . XXXVII . Evidence for lateral aggregation of rhodopsin molecules in phospholipase C-treated bovine photoreceptor membranes; Olive J et al.; Photoreceptor membranes derived from isolated bovine rod outer segments, are subjected to treatment with phospholipase C (Bacillus cereus) . This results in varying degrees of hydrolysis of the membrane phospholipids into diglycerides and water soluble phosphate esters without loss of rhodopsin . Electron microscopic observations of thin sections and freeze-fractured preparations indicate extrusion of diglycerides from the membranes and their coalescence to lipid droplets, beginning at 20% hydrolysis of phospholipids . After 90% hydrolysis of phospholipids membranous structures are still present . The rhodopsin is located in these structures, presumably in the form of two-dimensional lateral aggregates . This explains the cross-fracturing of the membranous structures, regularly observed upon freeze-fracturing of the phospholipase-treated photoreceptor membranes. Int J Pept Protein Res, 1978 May, 11(5), 340 - 4 Purification of milk-clotting protease from Bacillus mesentericus strain 76; Mesrob BK et al.; A two-step procedure for the isolation of pure enzyme from Bacillus mesentericus strain 76 is developed . Ion-exchange chromatography on CM-Sephadex-C50 is used as a second step after precipitation from ethanol . The pure enzyme preparation, obtained after gel filtration on Sephadex-G25 of the ion-exchange chromatographically separated product, possess the highest specific activity achieved . The homogeneity of the final preparation is proved by determination of the N-terminal amino acid (Arg) and by SDS-PAGDE (single disc). Mikrobiologiia, 1978 May-Jun, 47(3), 436 - 41 {Effect of exogenous factors on extracellular alkaline ribonuclease synthesis in Bacillus mesentericus}; Kapranova MN et al.; Bacillus mesentericus was found to assimilate nucleic acids as a source of nitrogen and phosphorus . Nucleic acids added to the medium as a source of nitrogen or phosphorus stimulated synthesis of ribonuclease . When washed bacterial cells were incubated for a short period of time in a fresh nutrient medium containing RNA, synthesis of RNAase was also induced . Synthesis of the enzyme was inhibited by high concentrations of chloramphenicol and actinomycin D, and stimulated by low concentrations of actinomycin D . Therefore, alkaline RNAase is an inducible enzyme which participates in the nutrition processes of bacteria. J Bacteriol, 1978 May, 134(2), 434 - 9 Evidence linking penicillinase formation and secretion to lipid metabolism in Bacillus licheniformis; Fishman Y et al.; The formation of penicillinase by cultures of Bacillus licheniformis was preferentially suppressed by cerulenin, an antibiotic known to specifically inhibit fatty acid synthesis in microorganisms . The effect was studied at cerulenin concentrations that had almost no effect on the rate of cell growth and overall protein synthesis, but that reduced the rate of {14C}acetate incorporation (by 50 to 70%), indicating partial inhibition of lipid synthesis . The levels of both the released enzyme (exopenicillinase) and its cell-bound precursor were reduced to the same extent (70% to 80%) . Enzyme formation was gradually resumed after the removal of cerulenin or the addition of a mixture of fatty acids prepared from lipids extracted from B . licheniformis . Reversal was less effective as the time interval between treatment with cerulenin and addition of fatty acids increased . We conclude that de novo synthesis of fatty acids is required for the formation of both the membrane-bound and extracellular penicillinase . Suppression of the membrane-bound enzyme is a likely consequence of the altered membrane (decreased lipid-to-lipid ratio and increased density) seen in cerulenin-treated preparations . The corresponding suppression of exopenicillinase is consistent with the view that it is derived from the membrane-bound form . A mechanism linking the general class of exportable proteins to specific aspects of lipid synthesis is discussed. Biokhimiia, 1978 May, 43(5), 865 - 71 {Isolation and some properties of restriction endonuclease from Bacillus amyloliquefaciens}; Sokolov NN et al.; A partially purified preparation of restriction endonuclease Bam I was isolated from the cells of Bacillus amyloliquefaciens . The purification procedure was a modification of a method described by Wilson and Young . Isolation and purification of the enzyme involved disruption of the cells by ultrasonication, treatment with streptomycin sulfate, fractionation by ammonium sulfate, chromatography on DEAE-cellulose and hydroxylapatite and rechromatography on DEAE-cellulose . Restrictase Bam I splits the linear double-chain DNA molecule of phage gamma into six fragments . The enzyme retained its stability under storage on the ice for 1,5--2 months in a Na- or K-phosphate buffer with beta-mercaptoethanol (10 mM). Appl Environ Microbiol, 1978 May, 35(5), 906 - 10 Inactivation of Bacillus thuringiensis spores by ultraviolet and visible light; Griego VM et al.; The inactivation of Bacillus thuringiensis spores and spores treated with two protectants, one proteinaceous and the other a commercial product, Shade, at wavelengths of the near-ultraviolet and visible spectra and at 254 nm is described . Determination of the inactivating wavelengths may be used to establish an efficient sunlight protective system for B . thuringiensis when used as a microbial insecticide. Proc Natl Acad Sci U S A, 1978 May, 75(5), 2088 - 92 Aminoacyl-tRNA synthetases: affinity labeling of the ATP binding site by 2', 3' -ribose oxidized ATP; Fayat G et al.; Homogeneous Escherichia coli methionyl-, isoleucyl-, tryptophanyl-, and phenylalanyl-tRNA synthetases and Bacillus stearothermophilus methionyl- and tyrosyl-tRNA synthetases are irreversibly inactivated by reaction of their active ATP sites with the 2',3'-dialdehyde derivative of ATP obtained by periodate oxidation . In each case, the amount of 14C-labeled dialdehyde derivative incorporated per molecule of inactivated enzyme appears consistent with the expected active stoichiometry of the synthetase . These results strongly support the presence, at the active site of the studied aminoacyl-tRNA synthetases, of a common residue, probably a lysine whose epsilon-NH2 group is known, from the work of others, to form a Schiff's base specifically with the 2',3'-dialdehyde derivatives of ribonucleotides. J Immunol, 1978 May, 120(5), 1532 - 6 Biochemical and functional characteristics of the plasma membrane of macrophages from BCG-infected mice; Edelson PJ et al.; A set of quantitative signs of activation, previously developed in studies of inflammatory peritoneal macrophages, has been applied to the study of immunologically stimulated peritoneal cells . Mice that are infected systemically with Bacillus Calmette Guerin (BCG) and then challenged locally with soluble mycobacterial antigens generate populations of cells that spread rapidly in culture, display an elevated pinocytic rate, ingest IgMC-coated sheep erythrocytes, and have diminished levels of 5'-nucleotidase activity . These effects depend on sensitization with live organisms, require secondary antigenic challenge, and develop on the same schedule as does effective cell-mediated immunity . The challenge is antigen specific and cannot be replaced by a nonspecific inflammatory stimulus . Thus, the characteristics of inflammatory macrophages that were previously defined are applicable to the study of immunologically mediated macrophage activation . Cells sharing this pattern of characteristics, whatever their mode of generation, are proposed to represent a distinct class of differentiated macrophages, as compared with the resident macrophage population. Can J Microbiol, 1978 May, 24(5), 537 - 43 Inhibition of RNA polymerase from Bacillus thuringiensis and Escherichia coli by beta-exotoxin; Johnson DE; The characteristics of exotoxin inhibition of deoxyribonucleic acid (DNA) dependent ribonucleic acid (RNA) polymerase isolated from Escherichia coli and Bacillus thuringiensis were investigated . RNA polymerase isolated from a variety of growth stages was partially purified and assayed using several different native and synthetic DNA templates, and exotoxin inhibition patterns were recorded for each . Although 8 to 20-h RNA polymerase extracts of E . coli retained normal sensitivity to exotoxin (50% inhibition at a concentration of 7.5 X 10(-6) M exotoxin), RNA polymerase isolated from late exponential and ensuing stationary-phase cultures of B . thuringiensis were nearly 50% less sensitive than exponential RNA polymerase activity . Inhibition patterns relating culture age at the time of RNA polymerase extraction to exotoxin inhibition suggested a direct correlation between diminishing exotoxin sensitivity and sporulation . Escherichia coli RNA polymerase could be made to mimic the B . thuringiensis exotoxin inhibition pattern by removal of sigma from the holoenzyme . After passage through phosphocellulose, exotoxin inhibition of the core polymerase was 30% less than the corresponding inhibition of E . coli holoenzyme . Heterologous enzyme reconstruction and assay were not possible due to loss of activity from the B . thuringiensis preparation during phosphocellulose chromatography, apparently from the removal of magnesium . In enzyme velocity studies, inhibition with exotoxin was noncompetitive with respect to the DNA template in the RNA polymerase reaction. J Bacteriol, 1978 May, 134(2), 389 - 93 alpha-amylase from five strains of Bacillus amyloliquefaciens: evidence for identical primary structures; Borgia PT et al.; The alpha-amylases from five strains of Bacillus amyloliquefaciens were compared to determine whether differences in primary structure are responsible for variations in catalytic properties previously reported among the enzymes . Amino acid analysis established virtually identical compositions for the proteins . Reaction with dimethylaminoaphthylene sulfonylchloride indicated the amino-terminal amino acid of each amylase to be valine . Carboxyl termini of the enzymes have been determined by digestion with carboxypeptidase A . The resulting kinetic data indicate tyrosine as the carboxyl terminus and leucine as the penultimate residue for all five proteins . Isoelectric focusing of the enzymes yielded isoelectric points in the pH range of 5.09 to 5.18 . Tryptic digests of the enzymes chromatographed on a cation-exchange column showed identical elution patterns . It is concluded that the primary structure of the amylase from the five strains is identical or exhibits only conservative substitutions. Cancer Res, 1978 May, 38(5), 1414 - 9 Induction of myeloid colony-stimulating activity in murine monocyte tumor cell lines by macrophage activators and in a T-cell line by concanavalin A; Ralph P et al.; Certain fibrosarcoma lines in culture and the WEHI-3 myelomonocytic leukemia cell line have previously been shown to secre