Eur J Biochem, 1975 Oct 15, 58(2), 453 - 60 Trypsin-kallikrein isoinhibitor K (type Kunitz) from snails (Helix pomatia) . Purification and characterization; Dietl T et al.; A basic proteinase inhibitor, isoinhibitor K, was purified by SE-Sephadex C-25 column chromatography from the mixture of acid-stable and heat-stable isoinhibitors of the snail (Helix pomatia) . Isoinhibitor K is homogeneous in polyacrylamide gel, cellulose acetate and polyacrylamide-dodecylsulfate electrophoresis . From the electrophoretic mobility in dodecylsulfate-polyacrylamide gel and apparent molecular weight of 6500 +/- 200 was estimated . From the amino acid composition the inhibitor consists of 58 amino acid residues . It contains three disulfide bridges, a C-terminal valine and a lysine residue at the reactive site . Isoinhibitor K inhibits the enzymes: bovine trypsin and chymotrypsin, porcine plasmin and pancreatic kallikrein, the trypsin-like component of Streptomyces griseus proteinase-pronase E, and fungi proteinase K from Tritirachium album Limber, which is only inhibited very slightly in contrast to the effect of the mixture of isoinhibitors . The inhibitory effect of isoinhibitor K against these enzymes is compared to that of the mixture or of other isoinhibitors . The following enzymes are not inhibited by isoinhibitor K: Aspergillus proteinase P and alkaline bacillus proteinase 2231 (Rohm), which both are inhibited by the mixture of isoinhibitors . Porcine elastase, bacterial proteinase N (M) (Rohm), and a trypsin-like proteinase from wheat are not inhibited, porcine acrosin and porcine serum kallikrein only to a very minor extent by the mixture of isoinhibitors . Reactive-site peptide-bond cleavage during inhibition could not be detected . Thus, the inhibitory behaviour is just as broad in specificity and as unusual as that of the trypsin-kallikrein inhibitor (Kunitz) from bovine organs . The N-terminus is blocked by pyroglutamic acid . Isoinhibitor K is the main component of the isoinhibitors secreted into the mucus and amounts to 35-40% of the mixture. Am J Clin Pathol, 1975 Oct, 64(4), 540 - 3 Bacteremia due to Succinivibrio dextrinosolvens . Report of a case; Southern PM Jr; An unusual case report of a patient with bacteremia due to Succinivibrio dextrinosolvens is presented . Heretofore, Succinivibrio spp . were thought to occur only as organisms in the rumens of herbivorous animals . Succinivibrio is an anaerobic, Gram-negative, curved, spiral bacillus with a polar monotrichous flagellar pattern . Volatile acids produced from glucose metabolism include succinic, acetic, formic and lactic . Various carbohydrate substrates are fermented to produce strong or weak acid end-points . Succinivibrio spp . will grow in bile, will not hydrolyze esculin or starch, and do not produce indol, catalase, lecithinase, lipase, or hemolysis . Bacteremia in this case was thought to have been the result of hypotension in the course of severe gastrointestinal hemorrhage . This is thought to represent the first published case of human infection due to Succinivibrio spp. J Biol Chem, 1975 Oct 10, 250(19), 7554 - 63 Substrate-protein interaction in tryptophanase from Bacillus alvei . Kinetic and spectral evaluations; Fenske JD et al.; This investigation studied the substrate protein interaction of the alpha, beta elimination reaction in tryptophanase (EC 4.1.99.1) . The results of this work are 2-fold . (a) The presence of multiple enzyme sites was found to be related to the observed kinetic patterns of inhibition . Indole analogues caused competitive inhibition in the tryptophanase reaction and noncompetitive inhibition in the dehydratase reaction . Inhibition patterns of alanine for these activities were reserved . (b) Under some conditions, compounds which bind presumably at the indole site modified the spectral and fluorescent characteristics of the enzyme . The addition of anthranilate to the enzyme resulted in a broad absorption band around 350 nm . This absorption band was distinct from that formed by alanine addition . Based on absorption data, both of these compounds could be bound simultaneously . The optical activity of tryptophanase was reported for the first time . Indole analogues caused greater conformational alterations in the circular dichroism spectra than 3-carbon analogues . The calculated anisotrophy factors, as well as fluorescent quenching data, suggest a more direct interaction between indole analogues and pyridoxal-P than between 3-carbon compounds that the coenzyme . It is proposed that the indole site is the dominant recognition site . The data are consistent with the three-dimensional aspects of space-filling models of Schiff's bases evaluated in terms of multiple site binding. Zh Mikrobiol Epidemiol Immunobiol, 1975 Oct, (10), 83 - 8 {Study of the process of interaction of the causative agent with the cells of the body and with a macrophage culture in experimental typhoid infection and carrier state}; Barshtein IuA et al.; Experiments were conducted on 117 rabbits and cells of the macrophage cultures in vitro by the methods of clinico-laboratory, quantitative microbiological, immunological, electron microscopic and microcinematographic examination; a study was made of the interaction of the typhoid causative agent with the cells of the organism and the macrophage cultures and also of some aspects of the immune response during acute typhoid infection and carrier state . Infection was modelled by the enteral, subconjunctival and intrabonemarrow infection with 24-hour culture of the typhoid bacillus (strain Ty2 4446) . Experiments demonstrated that structural reconstruction of both the causative agent and of the cells of the organism, of the culture macrophages and their organoids occurred in the course of the first hour after the infection . Homogenates of the lymphoid and myeloid tissues and also of the macrophages and polymorphonuclears possessed bactericidal activity against S . typhi . The degree of this activity largely depended on the pH of the medium . It was also shown that under conditions of the macrophage culture sodium aside inhibited the bactericidal activity of macrophages obtained from the intact and immune animals. Appl Microbiol, 1975 Oct, 30(4), 514 - 8 Efficacy of the inactivation of bacterial spores in white petrolatum and a hydrophilic ointment by gamma irradiation; Oie SH et al.; To evaluate the possibilities of using gamma irradiation for the sterilization of ointments, the effect of irradiation on spores of Bacillus pumilus and Bacillus sphaericus in dry material and in two different kinds of ointments was studied . The results indicate that for sterilization purposes irradiation was less effective in white petrolatum as compared to irradiation in the dry state . No such protective effect was found in a hydrophilic ointment . Accordingly, the sterilization dose needed for the sterilization of an ointment can be decided upon only after inactivation experiments with suitable test organisms in the actual preparation. Am J Vet Res, 1975 Oct, 36(10), 1545 - 7 Effect of aflatoxin on susceptibility of hamsters to Mycobacterium paratuberculosis; Larsen AB et al.; After oral administration of Mycobacterium paratuberculosis to hamsters, the organism passed the epithelial barrier of the intestine, and infection was established in the small intestine and mesenteric lymph nodes . The addition of aflatoxin to the ration of hamsters did not increase their susceptibility to M paratuberculosis but, rather, seemed to decrease susceptibility to the bacillus . Hamsters not treated with aflatoxin and infected with M paratuberculosis had higher bacterial counts in intestinal tract and mesenteric lymph node on necropsy than did infected hamsters that had been treated with aflatoxin . Aflatoxin-treated hamsters grew slowly, had an unthrifty appearance, and developed lesions of megalocytosis regardless of whether they were infected with M paratuberculosis. Eur J Biochem, 1975 Oct 1, 58(1), 185 - 92 Polyadenylation of RNA in vitro in isolated chromatin and nuclei; De Pomerai DI et al.; Poly(A) is added post-transcriptionally to RNA transcribed in vitro by endogenous form B DNA-dependent RNA polymerase bound to the template in isolated nuclei or chromatin . It is also added to processed fragments of the products from alpha-amanitin-resistant RNA polymerases A and/or C . The poly(A) segments are of similar size to those found in nuclear RNA pulse-labelled in vivo and are added onto the 3' terminus of RNA chains (whether pre-existing, completed during the incubation in vitro or created by fragmentation of larger RNA transcripts) . That poly(A) addition is not directly mediated by any of the nuclear DNA-dependent RNA polymerases is shown by the differential sensitivities of RNA and poly(A) syntheses to increasing ionic strength and transcriptional inhibitors such as the exotoxin from Bacillus thuringiensis. Acta Pathol Microbiol Scand {B}, 1975 Oct, 83(5), 513 - 8 The effect of bacitracin and Mn(II)ions upon the producer strain Bacillus licheniformis; Haavik HI; The peptide antibiotic bacitracin is inhibitory to growth of the producer strain Bacillus licheniformis only in the presence of excess manganese(II)ions . Both the early and the late growth are inhibited in a similar manner upon addition of bacitracin and manganese(II)ions . Thus, B . licheniformis does not develop resistance to its own antibiotic during growth . Added bacitracin is stimulatory to growth of B . licheniformis in media with a very low content of manganese(II)ions . These results support the hypothesis that bacitracin participates in the manganese transport of the producer strain B . licheniformis. Bull N Y Acad Med, 1975 Oct, 51(9), 1084 - 95 Antibiotics and endotoxic shock; McCabe WR; PIP: 2 topics of particular interest are addressed in this summary of data concerning antibiotics and endotoxic shock: 1) the antibiotic treatment of gram-negative bacillary infections complicated by shock differs from the treatment of similar infections not associated with shock; and 2) endotoxin per se is responsible for the occurrence of shock and other manifestation of gram-negative bacillary infections . With shock, inadequate tissue perfusion exists, requiring antibiotic administration be performed intravenously rather than by oral or intramuscular routes . In general, antibiotics act only to eradicate infection (i.e., kill bacteria) and in no way address the shock condition . Clinical results with polymyxins, implicated in some studies to neutralize endotoxin, assessed these drugs' clinical applicability by comparing the frequency of manifestations attributed to endotoxin, shock, and death occurring in patients with gram-negative bacteremia after treatment with polymyxin B or E . Treatment with 1 of the polymyxins failed to reduce the frequency of shock and death in gram-gegative bacteremia over that observed when other effective antibiotics were used initial therapy . Indeed, the frequency of complications was significantly greater in patients with ultimately fatal underlying diseases (P .05) and in all patients combined (P .001) . Human studies have attempted to define or pinpoint the relationship between endotoxin and disease symptoms (chills, fever, etc.), assuming an apparent correlation between the presence of circulating endotoxin and the frequency of fever, shock, or death compared with similar patients without detectable endotoxin; though the results did not preclude a role of endotoxin in symptomatology, they do cast considerable doubt on the widely held concept that circulating endotoxin is primarily responsible for the pathophysiologic changes observed during gram-negative infection . Using Limulus test data, the majority of studies found no connection between circulating endotoxin and presence of pathophysiologic changes . Factors affecting treatment of bacteremia are also discussed . Jpn J Exp Med, 1975 Oct, 45(5), 377 - 82 Tissue dispersion, cell harvest and fluid suspension culture by the use of bacterial neutral protease; Matsumura T et al.; Bacterial neutral protease of Bacillus polymyxa was found to disperse mammalian tissues and cells . Primary cell cultures were obtained from several tissues after treatments with 200 to 2,000 Kunitz unit per ml of this protease in either a phosphate buffer solution, a balanced salt solution or a tissue culture medium supplemented with serum . Monolayer cultures wither in their early passage levels or of established strains were harvested by a treatment with this protease, and proliferated again in monolayer after its removal . A growing culture of strain L-929 was kept in monodisperse suspension in the presence of this protease . In contrast to trypsin, this protease was found active in the presence of serum, stable during incubation and scarcely injured cells. Proc Natl Acad Sci U S A, 1975 Oct, 72(10), 4105 - 9 Comparison between macrophage activation and enhancement of nonspecific resistance to tumors by mycobacterial immunoadjuvants; Juy D et al.; It has repeatedly been observed that various bacterial preparations could increase the host's resistance to tumors . It has also been shown that after nonspecific activation by BCG (bacillus Calmette-Guerin), peritoneal macrophages could inhibit in vitro the growth of neoplastic target cells . In the present study a fraction extracted from Myobacterium smegmatis and referred to as interphase material was tested in view of measuring its ability to activate macrophages in vitro and in vivo . This preparation was previously shown to protect mice against a syngeneic leukemia and to increase the immune response of the guinea pig . Other water-soluble adjuvants devoid of demonstrable antitumor activity in vivo were also assayed . The results argue in favor of a correlation between adjuvant activity and the capacity of activating macrophages . Moreover, interphase material administered in vivo consistently induced stronger and more persistent stimulations of macrophages than the other preparations assayed. Proc Natl Acad Sci U S A, 1975 Oct, 72(10), 4037 - 41 Conditions controlling commitment of differentiation in Bacillus megaterium; Freese EB et al.; The developmental stage at which cells of Bacillus megaterium are committed to continue differentiation, i.e., sporulation, depends on both the previous growth medium and the new medium to which the cells are transferred for the commitment test . The latest "stage of no return," after which cells continue differentiation, no matter how rich in nutrients the medium, is reached as soon as the forespore is completely surrounded by a double membrane. Can J Microbiol, 1975 Oct, 21(10), 1464 - 7 Death rates of bacterial spores: nonlinear survivor curves; Han YW; Nonlinear survivor curves were obtained when spores of Bacillus cereus were heated in physiological saline solution . Curvilinear survivor curves did not appear to be caused by experimental artifacts but by the heterogeneity of spore population with regard to heat resistance. J Antibiot (Tokyo), 1975 Oct, 28(10), 764 - 9 The amino acid sequence of cerexin A (studies on antibiotics from the genus Bacillus . VII; Shoji J et al.; N-Bromosuccinimide cleavage reaction on cerexin A liberated allo-isoleucine . Treatment with conc . hydrochloric acid cleaved the antibiotic into two peptide fragments selectively at gamma-hydroxylysine residue . Deacylation with an enzyme preparation from Pseudomonas sp . afforded deacyl cerexin A . The amino acid sequences of these peptide fragments were examined by Edman degradation . From all the results, the entire amino acid sequence of cerexin A was deduced. Acta Pathol Microbiol Scand {B}, 1975 Oct, 83(5), 519 - 24 On the function of the polypeptide antibiotic bacitracin in the producer strain Bacillus licheniformis; Haavik HI; The growth of the bacitracin producing strain Bacillus licheniformis AL and the bacitracin-negative mutant SB 319 have been compared at different cultural conditions . Concentrations of the metal chelator EDTA which strongly inhibited the growth of the non-producer only slightly inhibited the growth of the bacitracin producer . The inhibitory effect of EDTA upon SB 319 was reversed by the addition of excess manganese(II)ions, cobalt(II)ions, or zinc(II)ions to the culture . The addition of several other ions had no such effect . The addition of bacitracin to the EDTA inhibited mutant also promoted growth . When the non-producer was mutated back to bacitracin production, the inhibitory effect of EDTA was lost . It is suggested that bacitracin may normally promote the uptake of several trace metals during growth of the producer organism. J Bacteriol, 1975 Oct, 124(1), 593 - 4 Protease associated with spores of Bacillus cereus; Tesone C et al.; A proteolytic activity is associated with the dormant spores of Bacillus cereus T and can be solubilized by washing the spores with 1 M KCl . This proteolytic activity is responsible for the attack of beta chains of ribonucleic acid-polymerase in extracts of dormant spores of this organism. J Bacteriol, 1975 Oct, 124(1), 542 - 9 Ribonucleic acid polymerase of germinating Bacillus cereus T; Hattori J et al.; It appears that a de novo synthesis of the deoxyribonucleic acid-dependent ribonucleic acid-polymerase in Bacillus cereus T takes place fairly late in outgrowth, at the onset of the vegetative cycle . Therefore, the ribonucleic acid-polymerase used by germinating spores is the one carried on from sporulating cells . However, the sporal enzyme is less soluble that the vegetative one, and its "core" is bound to two extra peptides . This complexing to other molecules could play a role in the regulation of gene expression during germination. Zentralbl Bakteriol {Orig B}, 1975 Oct, 161(2), 178 - 87 {Report of "lethal factor" synthetized by Bacillus cereus (author's transl)}; Balacescu C; Bacillus cereus growing in nutrition broth produces during logarithmic growth and in stationary phase a lethal factor extremely toxic for rodents . This toxin is only produced in the presence of oxygen and depends on bacterial replication . The highest titers of toxin are obtained using bacteria in a concentration of 10(7) to 10(9) per ml of nutrition broth . During regression phase of Bacillus cereus the titer of toxin declines to zero . Toxicity of the lethal factor becomes not altered in an alcaline or acidic medium . Temperatures exceeding 65 degrees C inactivate the toxin indicating its thermolability . Oral and rectal dispension of large quantities of the lethal factor does not induce toxic symptoms in rodents . However, when applicating toxin containing cultural filtrate of Bacillus cereus as an i.v., i.p., i.n . or s.c . injection it becomes highly toxic for mice, rats, hamsters, guinea pigs and rabbits . All animals injected by the different ways parenterally died. Biochem J, 1975 Oct, 151(1), 115 - 20 Poly(glucosylglycerol phosphate) teichoic acid in the walls of Bacillus stearothermophilus B65; Anderson AJ et al.; 1 . Walls of Bacillus stearothermophilus B65 contain a glycerol teichoic acid in which repeating structures consisting of 1-O-alpha-D-glucopyranosylglycerol phosphate are held together by phosphodiester linkage between the glycerol and glucose moieties of adjacent units . 2 . The walls are not agglutinated on incubation with concanavalin A, nor does the isolated teichoic acid form a precipitate with this lectin . 3 . No evidence was obtained of the presence of the glucosylated (1 leads to 2)-poly(glycerol phosphate) teichoic acid which has previously been reported to occur in walls of this bacterium. J Exp Med, 1975 Oct 1, 142(4), 887 - 902 Differential stimulation of murine lymphoma growth in vitro by normal and BCG-activated macrophages; Nathan CF et al.; Peritoneal macrophages from mice infected with Bacille Calmette-Guerin (BCG) and from normal mice were examined for their effects in vitro on thymidine uptake by 10 murine lymphomas, a murine fibroblast line, and a guinea pig hepatoma . Only the murine fibroblast line showed growth inhibition in the presence of BCG macrophages . For the majority of tumors, normal macrophages were profoundly stimulatory to tumor cell DNA synthesis, while BCG macrophages were much less stimulatory, without being frankly inhibitory . The effect of 2-mercaptoethanol on tumor cell growth was also studied . All lymphomas stimulated to grow more rapidly in vitro by normal macrophages were stimulated to a similar degree by 2-mercaptoethanol. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1975 Oct-Dec, 20(4), 239 - 43 {Protein changes in the jejunal juice and endointestinal exudation of albumin-I 131 in acute infectious enterocolitis}; Josan R et al.; The phenomenon of endointestinal protein exudation in acute infectious enterocolitis is studied . Total proteins were determined in 30 cases of acute enterocolitis and 50 of bacillary dysentery in the acute stages of the disease and convalescence . The proteinogram of the jejunal juice was performed in the acute stage and convalescence in 20 patients . In 16 patients and 5 controls endointestinal albumin elimination was determined quantitatively by means of 131I labeled albumin . The results showed increase in the total protein content in the jejunal juice in the course of acute infectious enterocolitis and bacillary dysentery and a return to normal values in convalescence . Electrophoresis of the jejunal juice in acute infectious enterocolitis showed the absence of fraction III with alpha1-globulin migration, and increased fractions I, II and IV probably due to the loss of endointestinal albumin, also confirmed by quantitative albumin determinations with 131I labeled albumin . In conclusion, patients with acute infectious enterocolitis present a marked loss of endointestinal albumins in the acute stage of the disease, with a return to normal values in convalescence. Int J Lepr Other Mycobact Dis . 1975 Oct-Dec;43(4):306. Mycobacterial antigens in antibody responses of leprosy patients; Kronvall G et al.; A reference system for M . smegmatis antigens in crossed immunoelectrophoresis was used to study antibody activities in serum samples of 91 leprosy patients . All polar and borderline lepromatous patients were positive . Mean numbers out of 14 M . smegmatis antigens involved were 4.3 and 3.5, respectively . Precipitins against antigen no . 1 were seen in all lepromatous cases . Antibodies against this antigen were detected in 50% of tuberculoid (polar, subpolar and borderline) cases . Antibody activity against M . avium and M . duvalii antigens was also detected using a staphylococcal radio-immuno-assay . Borderline and polar lepromatous cases showed elevated levels . Antigenic comparisons were made between four slow growing mycobacteria, fourteen fast growing mycobacteria and the leprosy bacillus using lepromatous serum pools as antibody reagents . Four of the antigens detected in M . leprae were also found in slow growing as well as fast growing species indicating a common occurrence among mycobacteria . Antigen no . 1 of M . duvalii, with an apparent molecular weight of 290,000, showed nonprotein characteristics . Further analysis of antigen no . 21, using lepromatous serum pools as antibody reagents, indicated the existence of at least two groups of antigenic determinants . In addition to determinants shared by all mycobacteria, there were antigenic structures apparently unique to M . leprae. Hoppe Seylers Z Physiol Chem, 1975 Oct, 356(10), 1613 - 23 D-glucose dehydrogenase from Bacillus megaterium M 1286: purification, properties and structure; Pauly HE et al.; 1) Glucose dehydrogenase from Bacillus megaterium has been purified to a specific activity of 550 U per mg protein . The homogeneity of the purified enzyme was demonstrated by gel electrophoresis and isoelectric focusing . 2) The amino acid composition has been determined . 3) The molecular weight of the native enzyme was found to be 116000 by gel permeation chromatography, in good agreement with the values of 120000 and 118000, which were ascertained electrophoretically according to the method of Hedrick and Smith and by density gradient centrifugation, respectively . 4) In the presence of 0.1% sodium dodecylsulfate and 8M urea, the enzyme dissociates into subunits with a molecular weight of 30000 as determined by dodecylsulfate gel electrophoresis . These values indicate that the native enzyme is composed of four polypeptide chains, each probably possessing one coenzyme binding site, which can be concluded from fluorescent titration of the NADH binding sites . 5) In polyacrylamide disc electrophoresis, samples of the purified enzyme exhibit three bands of activity, which present the native (tetrameric) form of glucose dehydrogenase and two monomeric forms (molecular weight 30000), arising under the conditions of pH and ionic strength of this method . 6) The enzyme shows a sharp pH optimum at pH 8.0 in Tris/HCl buffer, and a shift of the pH optimum to pH 9.0 in acetate/borate buffer . The limiting Michaelis constant at pH 9.0 for NAD is 4.5 mM and 47.5 mM for glucose . The dissociation constant for NAD is 0.69 mM . 7) D-Glucose dehydrogenase is highly specific for beta-D-glucose and is capable of using either NAD or NADP . The enzyme is insensitive to sulfhydryl group inhibitors, heavy metal ions and chelating agents. C R Acad Sci Hebd Seances Acad Sci D, 1975 Sep 29, 281(13), 953 - 6 {Demonstration of a single form of alpha-acetohydroxy acid synthetase in Bacillus cereus T}; Raimond J; Existence of one form of alpha-acetohydroxyacid synthetase shown by sucrose density gradient ultracentrifugation analysis, and inhibition in vitro by valine studies on the enzyme of Bacillus cereus T. J Biol Chem, 1975 Sep 25, 250(18), 7139 - 46 Reactivation of the lipid-depleted pyruvate oxidase system from Escherichia coli with cell envelope neutral lipids; Cunningham CC et al.; The pyruvate oxidase system of Escherichia coli is composed of a soluble flavoprotein, pyruvate oxidase (EC 1.2.2.2, pyruvate:ferricytochrome b1 oxidoreductase), and an electron transport system associated with the cell envelope-membrane fraction . The membrane particles contain 15% lipid by weight . Fractionation of the lipids revealed that abut one-third are neutral lipids and two-thirds are phospholipids . The relative ratio of ubiquinone to menaquinone within the neutral lipid fraction is 15:1 on a molar basis . Removal of the lipids from the membrane particles by extraction with aqueous acetone or hydrolysis of the phospholipids by treatment with Bacillus cereus phospholipase C results in a complete loss of electron transport activity . Analysis of the particles extracted with aqueous acetone revealed that practically all the neutral lipids and 65% of the phospholipids are removed by this treatment . Phospholipase treatment results in a loss of 75% of the membrane phospholipid phosphorus; however, the diglycerides and the neutral lipids produced by phospholipase hydrolysis remain associated with the particles . Addition of neutral lipid and a detergent, hepta-DL-alanyl dodecylamide to the acetone-extracted material results in a restoration of 37% of the original particle activity . Addition of neutral lipid and hepta-DL-alanyl dodecylamide to phospholipase-treated particles completely restores the original electron transport activity . Furthermore, addition of ubiquinone from either yeast (UQ6) or E . coli (UQ8) will restore pyruvate oxidase activity when the quinones are supplemented with photoinactivated neutral lipid . No restoration of activity to phospholipase-treated particles is noted upon the addition of either menaquinone 6 or menaquinone 8 to the reconstitution system . In fact, these compounds appear to suppress restoration of activity when they are added to reaction mixtures containing neutral lipid and phospholipase-treated particles. Biochemistry, 1975 Sep 23, 14(19), 4298 - 304 Structural role of pyridoxal 5'-phosphate, pyridoxal 5'-phosphate analogs, and other agents in the association of subunits of Bacillus alvei apotryptophanase; Isom HC et al.; Bacillus alvei apotryptophanase readily dissociates at low protein concentration and sediments at 5.7 S (dimer) in 0.01 M potassium phosphate (pH 7.8) from 9 to 33 degrees . With temperature held constant at 9 degrees, increasing the potassium, sodium, or ammonium phosphate buffer concentration increases the sedimentation value to 8.0 S . Increasing the monovalent cation concentration alone does not have the effect . Imidazole and pyridoxal compete with phosphate, preventing the effect . Raising the temperature to 26 degrees in the presence of high concentrations of potassium phosphate increases the sedimentation constant to 9.4 S . The addition of pyridoxal-P converts the dimer to a 9.4S tetramer . The conversion is dependent upon coenzyme concentration, temperature, and the nature of monovalent cation present . The Km for pyridoxal-P for the sodium form of the enzyme is more than tenfold greater than the Km for the potassium form of the enzyme . 2'-Methyl, 2'-hydroxyl, 6-methyl, and the N-oxide of pyridoxal-P are active in the association of dimer to tetramer but to differing extents . Analogs altered in the 4'-formyl position are also inactive structurally . Anthranilic acid, a competitive inhibitor of tryptophan, and 8-anilino-1-naphthalenesulfonic acid (ANS), a competitive inhibitor of pyridoxal-P binding, are both active in affecting the dimer to tetramer association but tryptophan is not . The dimer and tetramer are spectrally distinguishable through circular dichroic measurements, fluroescence quenching with pyridoxal-P or pyridoxal, and fluorescence enhancement with ANS . Pyridoxal-P causes the release of ANS from an ANS-apoenzyme complex. Biochemistry, 1975 Sep 23, 14(19), 4291 - 7 Pyridoxal 5'-phosphate and analogs as probes of coenzyme-protein interaction in Baccillus alvei tryptophanase; Isom HC et al.; Trytophanase from Bacillus alvei was resolved from its coenzyme, pyridoxal phosphate, by treatment with cysteine followed by column chromatography . Spectrophotometric titration of apoenzyme with pyridoxal-P showed 1 mol of pyridoxal-P bound per 52,000 g of enzyme . Kinetic analysis of coenzyme binding showed hyperbolic activation curves with a Km of 1.6 muM . Pyridoxal-P was used as a natural active site probe in spectrophotometric studies to distinguish differences in the active center of holotryptophanase and reconstituted enzyme that were not apparent by other techniques . The pKa for holotryptophanase is 7.9 while the pKa for reconstituted apoenzyme is 8.4 . Apotryptophanase binds 2-nor, 2'-methyl, 2'-hydroxy, 6-methyl, and N-oxide pyridoxal-P to form analog enzymes distinguishable on the basis of absorption spectra and relative activity in catalyzing both the alpha, beta-elimination and beta-replacement reactions of tryptophanase . Apoenzyme also binds pyridoxal but pyridoxal analog enzyme is not active. Klin Wochenschr, 1975 Sep 15, 53(18), 881 - 3 Mechanisms by which tumors avoid destruction by the immune system BCG-catalyzed increase in IgG and IgA blocking activity of lymphocyte-mediated cytotoxicity; Hakim AA; The immunoglobulin IgM fraction from the serum of one week sarcoma-bearing BALB/c mice increased, the IgG and IgA fractions from the same animal had no effect on the lymphocytemediated cytotoxic release of 51Cr from 51Cr-labelled sarcoma cells . These latter two immunoglobulin fractions from serum of 14 Days, or more, sarcoma-bearing mice inhibited the lymphocyte-mediated cytotoxic activity . The IgA, IgG or IgM fractions from mice inoculated with bacillus Calmette-Guerin (BCG) had no effect, but if these same animals were inoculated with viable sarcoma cells (10(2) to 10(6) cells per mouse) IgG and IgA fractions inhibited the lymphocyte-mediated cytotoxic activity . The magnitude of inhibition was greatest with sera or IgG from tumor-bearing BCG-treated animals. C R Acad Sci Hebd Seances Acad Sci D, 1975 Sep 15, 281(11), 755 - 8 {A Bacillus thuringiensis Berliner mutant resistant to oxytetracycline, with temperature-sensitive sporulation}; Fargette F et al.; Sporulation of an oxytetracycline-resistant mutant is blocked at 37 degrees C and delayed at 30 degrees C; shift up and shift down (30-37 degrees C) and assays of some extra- and intracellular enzymes show that an early event (stage II) is concerned. Biochim Biophys Acta, 1975 Sep 12, 407(1), 61 - 72 On the nature of the cytosine-methylated sequence in DNA of Bacillus brevis var . G.-B; Vanyushin BF et al.; On growing the cells of Bacillus brevis S methionine-auxotroph mutant in the presence of {Me-3H}methionine, practically all the radioactivity incorporated into DNA is found to exist in 5-methylcytosine and N6-methyladenine . The analysis of pyrimidine isopliths isolated from DNA shows that radioactivity only exists in mono- and dinucleotides and the content of 5-methylcytosine in R-m5 C-R and R-m5 C-T-R oligonucleotides is equal . The analysis of dinucleotides isolated from DNA by means of pancreatic DNAase hydrolysis allows the nature of purine residues neighbouring 5-methylcytosine to be identified and shows that 5-methylcytosine localizes in G-m5 C-A and G-m5 C-Tr fragments . B . brevis S DNA methylase modifying cytosine residues recognizes the GCA/TGC degenerate nucleotide sequence which is a part of the following complementary structure with a two-fold rotational axis of symmetry: (5')...N'-G-C-T-G-C-N.. . (3') (3')...N-C-G-A-C-G-N'.. . (5') (Methylated cytosine residues are askerisked) . Cytosine-modifying DNA methylase activity is isolated from B . brevis cells; it is capable of methylating in vitro homologous and heterologous DNA . Hence DNA in bacterial cells can be undermethylated . This enzyme methylates cytosine residues in native and denatured DNA in the same nucleotide sequences . Specificity of methylation of cytosine residues in vitro and in vivo does not depend on the nature of substrate DNA . DNA methylases of different variants of B . brevis (R, S, P+, P-)) methylate cytosine residues in the same nucleotide sequences . It means that specificity or methylation of DNA cytosine residues in the cells of different variants of B . brevis is the same. J Biol Chem, 1975 Sep 10, 250(17), 6983 - 9 D-amino acid aminotransferase of Bacillus sphaericus . Enzymologic and spectrometric properties; Yonaha K et al.; D-Amino acid aminotransferase, purified to homogeneity and crystallized from Bacillus sphaericus, has a molecular weight of about 60,000 and consists of two subunits identical in molecular weight (30,000) . The enzyme exhibits absorption maxima at 280, 330, and 415 nm, which are independent of the pH (5.5 to 10.0), and contains 2 mol of pyridoxal 5'-phosphate per mol of enzyme . One of the pyridoxal-5'-P, absorbing at 415 nm, is bound in an aldimine linkage to the epsilon-amino group of a lysine residue of the protein, and is released by incubation with phenylhydrazine to yield the catalytically inactive form . The inactive form, which is reactivated by addition of pyridoxal 5'phosphate, still has a 330 nm peak and contains 1 mol of pyridoxal 5'-phosphate . Therefore, this form is regarded as a semiapoenzyme . The holoenzyme shows negative circular dichroic bands at 330 and 415 nm . D-Amino acid aminotransferase catalyzes alpha transamination of various D-amino acids and alpha-keto acids . D-Alanine, D-alpha-aminobutyrate and D-glutamate, and alpha-ketoglutarate, pyruvate, and alpha-ketobutyrate are the preferred amino donors and acceptors, respectively . The enzyme activity is significantly affected by both the carbonyl and sulfhydryl reagents . The Michaelis constants are as follows: D-alanine (1.3 and 4.2 mM with alpha-ketobutyrate and alpha-ketoglutarate, respictively), alpha-ketobutyrate (14 mM withD-alanine), alpha-ketoglutarate (3.4 mM with D-alanine), pyridoxal 5'-phosphate (2.3 muM) and pyridoxamine 5'-phosphate (25 muM). Appl Microbiol, 1975 Sep, 30(3), 489 - 92 Gamma-aminobutyric acid pathway and modified tricarboxylic acid cycle activity during growth and sporulation of Bacillus thuringiensis; Aronson JN et al.; Enzymatic analyses of Bacillus thuringiensis extracts suggest that a modified Krebs tricarboxylic acid cycle (without alpha-ketoglutarate dehydrogenase) can operate during sporulation in conjunction with the glyoxylic acid cycle and the gamma-aminobutyric acid pathway. J Exp Zool, 1975 Sep, 193(3), 361 - 7 DNA-RNA bodies in midgut cells of the stick insect, Bacillus rossius; Scali V et al.; In the anterior part of the midgut and in the Malpighian tubules of the stick insect Bacillus rossius, about 10% of the epithelial cells develop endonuclear bodies which appear as DNA-RNA masses; in these cells the usual nucleoli are no longer evident . The DNA-RNA bodies are first formed in third instar larvae, become numerous in the fourth instar and persist in adults . In all larval instars and adults a different kind of DNA-body has been noticed in the epithelial cells of the posterior midgut . The DNA-RNA bodies of the anterior midgut and of the Malpighian tubules have been interpreted as the result of somatic gene amplification, whereas the DNA masses of the posterior midgut are likely due to a virus infection. J Bacteriol, 1975 Sep, 123(3), 806 - 14 Evidence for extrusion of unfolded extracellular enzyme polypeptide chains through membranes of Bacillus amyloliquefaciens; Sanders RL et al.; The production of extracellular alpha-amylase and protease by protoplasts of Bacillus amyloliquefaciens has been achieved . The production of enzymically active protease was totally dependent on a high concentration of either Mg2+, Ca2+, or spermidine, but production of active alpha-amylase was not . This cation dependence of protease production was seen immediately upon addition of lysozyme to intact cells . The cations could prevent the inactivation of protease and alter the cytoplasmic membrane configuration of protoplasts . Production of active alpha-amylase and protease by protoplasts was totally inhibited by proteolytic enzymes such as trypsin, alpha-chymotrypsin, or the organism's purified extracellular protease . The evidence suggests that these degradative enzymes act specifically on the emerging polypeptide of the extracellular enzyme and that the polypeptide emerges in a conformation different from that of the native molecule. Proc Natl Acad Sci U S A, 1975 Sep, 72(9), 3463 - 7 Degradation of penicillin G to phenylacetylglycine by D-alanine carboxypeptidase from Bacillus stearothermophilus; Hammarstrom S et al.; D-Alanine carboxypeptidase from Bacillus stearothermophilus is a membrane-bound enzyme which is inhibited by covalent interaction with penicillin G . The penicilloyl enzyme spontaneously reactivates and simultaneously releases a penicillin G degradation product; 0.2 mumol of the latter was isolated after incubation of 4.2 mumol of {8-14C}penicillin G with 10 g of membrane protein . It was identified as phenylacetylglycine by chromatographic techniques, infrared spectroscopy, and mass spectrometry . A mechanism for the degradation is proposed in which the remaining part of penicillin G would be released as 5,5-dimethyl-delta2-thiazoline-4-carboxylic acid . The implications of this finding are discussed. Appl Microbiol, 1975 Sep, 30(3), 445 - 9 Liquid nitrogen cryo-impacting: a new concept for cell disruption; Smucker RA et al.; High-efficiency disruption of bacteria can be accomplished in 2 or more min by the new procedure of liquid nitrogen cryo-impacting . Release of the dipicolinic acid-Ca2+ chelate paralleled the breakage of Bacillus megaterium endospores . Lactate dehydrogenase activity was much better in supernates from liquid nitrogen cryo-impacting-broken Escherichia coli cells than in those from sonically treated and broken E . coli cells. Appl Microbiol, 1975 Sep, 30(3), 439 - 44 Degradation of 3-hydroxybenzoate by bacteria of the genus Bacillus; Crawford RL; The pathway whereby certain bacterial strains of the genus Bacillus degrade m-hydroxybenzoate is delineated . Of 12 strains examined, nine were tentatively classified as representatives of the species Bacillus brevis, two of Bacillus sphaericus and one of Bacillus megaterium . All strains degraded m-hydroxybenzoate via the same pathway . m-Hydroxybenzoate was hydroxylated to 2,5-dihydroxybenzoate (gentisate), which was oxidized by a gentisate 1,2-deoxygenase yielding maleylpyruvate . Maleylpyruvate was hydrolyzed without prior cis, cis to cis, trans isomerization yielding pyruvate and maleic acid . Numerous soils were examined by plate-count procedures and found to contain 10(4) to 10(6) aerobic sporeformers able to grow on m-hydroxybenzoate per g of dry soil. Eur J Biochem, 1975 Sep 1, 57(1), 221 - 30 Terminal-sequence analysis of bacterial ribosomal RNA . Correlation between the 3'-terminal-polypyrimidine sequence of 16-S RNA and translational specificity of the ribosome; Shine J et al.; The 3'-terminal sequences of 16-S ribosomal RNA from a number of bacteria have been determined by a stepwise degradation and 3'-terminal labelling procedure . The sequences obtained were: Bacillus stearothermophilus, -G(Z)approximately 5 Y-U-C-C-U-U-U-C-U (A); B . subtilis, -G(Z)approximately 7 Y-C-U-U-U-C-U; Caulobacter crescentus, -G(Z)3 Y-U-C-C-U-U-U-C-U; Pseudomonas aerugionosa, -G-Z-Z-Y-C-U-C-U-C-C-U-U(A), where Z is any nucleotide other than G . Thus, as previously found in Escherichia coli, all bacterial 16-S rRNAs contain a pyrimidine-rich tract at the 3'-terminus . In B . stearothermophilus and Ps . aeruginosa this region shows substantial heterogeneity involving the 3'-terminal adenylic acid . A low level of 3'-terminal heterogeneity cannot be excluded for the other bacterial 16-S rRNAs examined . The 3'-termini of bacterial 16-S rRNA can be divided into two groups on the basis of sequence homology . The first group comprises E . coli and Ps . aeruginosa; the second, B . stearothermophilus, B . subtilis and C . crescentus . This division correlates with a previous separation of bacterial ribosomes into two categories based on ability to translate different mRNA preparations {Stallcup, Sharrock & Rabinowitz (1974) Biochem . Biophys . Res . Commun . 58, 92-98} . We have previously proposed that the precise base sequence at the 3'-terminus of 16-S rRNA determines the intrinsic capacity of bacterial ribosomes to translate a particular cistron {Shine & Dalgarno (1975) Nature (Lond.) 254, 34-38} . No difference was found in the 3'-terminal heptanucleotide sequence of 16-S rRNA from bacteriophage T7-infected E . coli, as compared to that in uninfected cells . Thus, the T7-induced alteration in translational specificity of E . coli ribosomes is probably not mediated by modification of the terminal seven nucleotides of the smaller rRNA . The 3'-terminal sequences of the 23-S rRNA species were also determined . The sequences obtained were: B stearothermophilus and B . subtilis, -Y-C; C . crescentus, -Y-C-U; Ps . aeruginosa, -Y-C-A; E . coli, -G-Y-U-U-A-A-C-C-U-U . No evidence for 3'-terminal heterogeneity was found . The results obtained are discussed in relation to possible base-pairing roles for the 3'-end of 16-S rRNA in bacterial protein synthesis. J Bacteriol, 1975 Sep, 123(3), 1197 - 207 Cell wall growth of Bacillus megaterium: cytoplasmic radioactivity after pulse-labeling with tritiated diaminopimelic acid; de Chastellier C et al.; Study of the cell wall growth in Bacillus megaterium by pulse-labeling a DAP- Lys- mutant with tritiated diaminopimelic acid (DAP) had revealed the presence of intracytoplasmic radioactivity . The nature of this radioactivity was studied on one hand by autoradiographic analysis of bacteria treated in different ways and on the other hand by chromatography of the radioactive compounds extracted with boiling water . It is shown that cytoplasmic radioactivity corresponds neither to free DAP nor to DAP metabolized into lysine, but to murein precursors . Autoradiographic analysis of bacteria in which all murein precursors were removed gives exactly the same cell wall growth pattern as the one previously obtained for untreated bacteria . It can be concluded that, in B . megaterium, cell wall elongation occurs by diffuse intercalation of newly synthesized murein along the cylindrical part of the cell wall and that only cross wall formation occurs in a precise growth zone. J Bacteriol, 1975 Sep, 123(3), 1184 - 96 Study of cell wall growth in Bacillus megaterium by high-resolution autoradiography; de Chastellier C et al.; Growth of the cell wall of Bacillus megaterium was studied by pulse-labeling the cell wall of a DAP- Lys- mutant for a very short time with tritium-labeled diaminopimelic acid . The distribution of radioactivity along the cell wall was examined by high-resolution autoradiography on isolated cell walls and thin sections of bacteria . The results indicate that cell wall elongation occurs by diffuse intercalation of newly synthesized murein into the expanding cell wall during exponential growth, as well as during germination, and that the only zone of highly localized diaminopimelic acid incorporation is found at the cross wall during its synthesis . This zone contains about 30% of the radioactivity incorporated into the cell wall . Analysis of autoradiographs of thin sections of bacteria shows that the total radioactivity incorporated per bacterium doubles during the life cycle . This doubling occurs in the cylindrical part of the cell wall but not in the polar caps . This seems to indicate that elongation of the bacterium is not constant during the life cycle but increases with the length of the cell. Appl Microbiol, 1975 Sep, 30(3), 354 - 61 Metalloprotease from Bacillus thuringiensis; Li E et al.; Bacillus thuringiensis var . kurstaki was shown to produce an extracellular, metal chelator-sensitive protease during the early stages of sporulation . Protease production in nutrient broth was dependent upon supplementation with Mn2+ or Ca2 . The addition of Ca24 was required for enzyme stabilization... Mikrobiologiia, 1975 Sep-Oct, 44(5), 791 - 4 {Change in the activity of the enzymatic systems of Bacillus anthracoides spores during germination and under the action of Ca hypochlorite}; Bekhtereva MN et al.; The activity of the enzymes of the citric acid cycle, glycolysis, and hexose monophosphate pathway was studied during germination of the spores of Bacillus anthracoides and upon their treatment with calcium hypochlorite . No activity of isocitrate dehydrogenase and glucose-6-phosphate dehydrogenase was found in the extracts of the vegetative cells, contrary to the spores and initiated spores . The activity of other enzymes changes but slightly in the course of germination of the spores . Treatment of the spores with calcium hypochlorite inhibited their initiation and germination and the activity of several enzymes, especially malate dehydrogenase, isocitrate dehydrogenase, and fumarase. Proc Natl Acad Sci U S A, 1975 Sep, 72(9), 3656 - 60 Enhancement of immunity against murine syngeneic tumors by a fraction extracted from non-pathogenic mycobacteria; Lamensans A et al.; The data reported here demonstrate that a preparation extracted from nonpathogenic mycobacteria such as Mycobacterium smegmatis and hereafter referred to as interphase material protected mice against Ehrlich ascitic carcinoma, L-1210 leukemia, and another syngeneic lymphoid leukemia . Furthermore, mice treated by this preparation were much less susceptible to endotoxins than when stimulated by BCG (bacillus Calmette-Guerin) or M . smegmatis cells . Moreover, guinea pigs treated by interphase material administered in Freund's incomplete adjuvant showed an increased immune response, yet their sensitivity to tuberculin was much weaker than that of controls sensitized with Freund's complete adjuvant . Finally, resistance to Columbia SK virus infection could be demonstrated when interphase material was administered to mice prior to virus challenge. Prikl Biokhim Mikrobiol, 1975 Sep-Oct, 11(5), 742 - 5 {Effect of the method of cell disintegration on the aspartate kinase activity of preparations from Bacillus polymyxa var . Ross.}; Chigaleichuk AG et al.; The influence of methods of cell disintegration of Bac . polymyxa on aspartate kinase activity (EC 2.7.2.4) was examined . The disruption by means of the Hews press yielded a more active preparation as compared with ultrasonic disintegration . The supernatant treatment with streptomycin sulphate increased the preparation activity 2-fold . Dialysis of the fraction with a high activity of aspartate kinase inactivated the enzyme by 80--85% . The relationship between the aspartate kinase activity and the portein and ATP concentration in the reaction mixture was established. Can J Microbiol, 1975 Aug, 21(8), 1270 - 2 Soil bacteriostasis: inhibition of spore germination and microcolony development in agar discs incubated on nonsterile soils; Davis RD; An agar-disc method showed that bacterial spores were subject to a bacteriostatic effect exerted by soil . Soil either inhibited spore germination or reduced microcolony development of the six Bacillus strains tested . Bacteriostasis was relieved by the addition of nutrients to soil-exposed discs after they had been removed from the soil. Can J Biochem, 1975 Aug, 53(8), 827 - 33 Carboxyl-terminal sequences of procaryotic ribosomal proteins from Escherichia coli, Bacillus stearothermophilus, and Halobacterium cutirubrum; Duggleby RG et al.; The carboxyl-terminal amino acid sequences of two ribosomal proteins, Escherichia coli L12 and E . coli S4, and the proteins believed to be their equivalents from Bacillus stearothermophilus and Halobacterium cutirubrum, were determined . These proteins are known to be required for peptide chain termination (L12) and in ribosome assembly (S4) . The carboxyl-terminal sequences obtained suggest that the E . coli and B . stearothermophilus proteins have retained structural homology in this region, whereas the H . cutirubrum proteins have not. Eur J Biochem, 1975 Aug 1, 56(1), 15 - 22 Role of ribosomal protein S1 in portein synthesis: effects of its addition to Bacillus stearothermophilus cell-free system; Isono S et al.; Products of the f2 phage RNA-directed protein synthesizing systems in vitro, derived from Escherichia coli and Bacillus stearothermophilus, were analyzed . In contrast to other reports, it was found that B . stearothermophilus ribosomes synthesized phage coat protein when the high-speed (100 000 X g) supernatant (S100) and ribosomal wash (crude initiation factors) of homologous origin were used . Furthermore, a marked stimulation of coat protein synthesis was observed with B . stearothermophilus ribosomes when either S100 or ribosomal wash or both from E . coli cells were used instead of the respective homologous components . The principle responsible for this stimulation was identified as the 30-S ribosomal protein S1 present in S100 and ribosomal wash . Purified S1 from E . coli at roughly one-to-one molar ratio to ribosomes resulted in about 10-fold stimulation of the incorporation of {14C}valine when added to the B . stearothermophilus system . This stimulation was observed mainly in the synthesis of coat protein and replicase. J Lab Clin Med, 1975 Aug, 86(2), 349 - 59 Radioimmunoassay, acetylating radio-enzymatic assay, and microbioassay of gentamicin: a comparative study; Stevens P et al.; Gentamicin is an aminoglycoside antibiotic widely used to treat gram-negative bacillary infections . Because it has a low therapeutic index, monitoring of serum levels may help to insure adequacy of dosage and avoid toxicity . Microbiological assays are relatively slow and can be complicated by the presence of other antimicrobials . Radioimmunoassay (RIA) and acetylating radio-enzymatic assay (ARA) are new methods for gentamicin assay which offer the following advantages: rapidity (less than 3 hours); no interference by other antibiotics; RIA is extremely sensitive and ARA is versatile (being useful in the measurement of other aminoglycosides) . Correlation coefficients determined by linear regression analysis of assays on 36 patient samples performed in duplicate on 2 different days demonstrated no significant difference in measurement of gentamicin by each of the methods . Factors such as numbers of specimens, cost, and time involved will affect the decision of the method to be applied in individual laboratories. Int J Radiat Biol Relat Stud Phys Chem Med, 1975 Aug, 28(2), 165 - 76 Two components in the radiation sensitization of bacterial spores by p-nitroacetophenone: the -OH component; Ewing D; p-Nitroacetophenone (PNAP) sensitizes Bacillus megaterium spores under anoxic conditions to the lethal effects of 50 kVp X-rays . Concentrations between approximately 5 X 10(-4) M and 3-8 X 10(-3) M produce the maximum effect, an increase of about 30 per cent over the anoxic response when the spores are irradiated in water . Compounds that scavenge -OH decrease, but cannot completely eliminate, this maximum amount of sensitization . These results indicate that PNAP acts to increase spores' radiation sensitivity through two separable types of chemical reactions: one which involves -OH and one which does not . Possible mechanisms responsible for these two components of damage are discussed . In these experiments 1/15 M phosphate buffer acts in several unexpected ways . This concentration itself increased the anoxic spore response by about 9 per cent (relative to the anoxic response in water) . In addition, although the maximum amounts of sensitization were the same, the amounts of sensitization from lower PNAP concentrations differed when the suspending fluid was buffer instead of water . An interaction was also seen during the PNAP-t-butanol experiments; again, the responses at low PNAP concentrations were different in buffer and in water . No mechanisms for these actions of this buffer were suggested, although somewhat similar effects may occur with other organisms . Clearly, such effects must be recognized and evaluated before quantitative analyses of the actions of sensitivity-modifying agents can be completed. Can J Microbiol, 1975 Aug, 21(8), 1192 - 7 Germination of Bacillus cereus endospores: a proposed role for heat shock and nucleosides; Yousten AA; Spores of Bacillus cereus T prepared in a glucose - yeast extract - salts broth germinated in L-alanine or more rapidly in L-alanine plus inosine or adenosine . The nucleosides alone were not germinative . Inosine was shown to produce no pregerminative changes in spores that prepared them for more rapid germination later in L-alanine . Experiments which measured the interaction of nucleosides, heat shock, and D-alanine on germination rates suggested that nucleosides may potentiate L-alanine-induced germination by causing discrimination against D-alanine at the L-alanine binding site(s) in the spore . D-Alanine is germinative when used with inosine probably because of L-alanine formation by alanine racemase . Heat shock, a prerequisite to D-alanine plus inosine-induced germination, may facilitate entry of inosine into the spore in amounts needed to discriminate against D-alanine. J Bacteriol, 1975 Aug, 123(2), 717 - 23 Passage of a membrane protein through the walls of toluene-treated Bacillus megaterium cells; Fan DP et al.; Based on autoradiographic and microscopic evidence, it seems likely that a membrane protein essential for peptidoglycan synthesis can be extracted from uhlysed toluene-treated Bacillus megaterium cells . Furthermore, this protein can be added back to the membrane through the wall to reconstitute peptidoglycan synthesis . Autoradiograms also show that peptidoglycan is synthesized from externally added nucleotide precursors over the entire length of the toluene-treated bacterial . The amounts of peptidoglycan made is to small to be visible by thin section electron microscopy. J Bacteriol, 1975 Aug, 123(2), 598 - 603 Lipid metabolism during bacterial growth, sporulation, and germination: an obligate nutritional requirement in Bacillus thuringiensis for compounds that stimulate fatty acid synthesis; Nickerson tkw et al.; The regulation of fatty acid biosynthesis by compounds that are required for growth of Bacillus thuringiensis was investigated using an vivo assay developed to measure fatty acid synthesis in germinating spores . A minimal glucose-ammonium-salts medium does not support growth even though previous radiorespirometric studies have shown B . thuringiensis to possess intact tricarboxylic acid and Embden-Meyerhof-Parnas pathways . Abundant growth does occur, however, when this medium is supplemented with either glutamate, aspartate, citrate, thiosulfate, cystine, or ethylenediaminetetraacetic acid . Cells held under nongrowing conditions incorporate acetate into fatty acids; fatty acid synthesis is stimulated by the compounds that permit growth . These alternate nutritional requirements are not manifestations of a vitamin or trace metal deficiency and do not reflect a chelation phenomenon . These results indicate a direct correlation between the capacity of these compounds to promote growth and to stimulate formation of fatty acids. J Bacteriol, 1975 Aug, 123(2), 463 - 70 Germination and peptidoglycan solubilization in Bacillus megaterium spores; Hsieh LK et al.; During initiation of Bacillus megaterium QM B1551 spore germination, trichloroacetic acid-soluble, nondialyzable peptidoglycan fragments with an average molecular weight of 20,000 were excreted . This solubilization of peptidoglycan was measured in vitro as the amount of trichloroacetic acid-soluble hexosamine released from a suspension of broken spores . HgC12, a potent inhibitor of initiation, had no effect on the in vitro solubilization of peptidoglycan . In vivo, HgC12 had no effect on peptidoglycan release from spores that had lost heat resistance, but HgC12 did block complete absorbance loss . These results suggest that mercury inhibits some reactions that normally occur before loss in heat resistance but not the subsequent peptidoglycan release, and mercury inhibits other reactions involved with complete absorbance loss. Can J Microbiol, 1975 Aug, 21(8), 1236 - 46 Biological characteristics of an enterotoxin produced by Bacillus cereus; Spira WM et al.; An enterotoxin synthesized during exponential growth by Bacillus cereus produces fluid accumulation in rabbit ileal loops, alters vascular permeability in the skin of rabbits, and kills mice when injected intravenously . All activities are eluted simultaneously from a Sephadex G-75 column and are distinct from the hemolysin and egg yolk turbidity factor of B . cereus . The enterotoxin is a true exotoxin . It interacts with intestinal receptor sites in a highly transient manner in the ileal loop system . Rabbit immune serum produced against the culture fluids from one strain of B . cereus neutralized the three biological activities in all other strains tested except strain B-6-ac for which none of the activities were neutralized . Enterotoxin proved to be unstable under a wide variety of conditions; ionic strength was especially critical . Enterotoxin was most stable in a pH range of 5.0 to 10.0, but lost activity rapidly outside this range . Alkylation provided some protection of enterotoxin activity in crude preparations but failed to protect activity during purification procedures . It did not appear to affect critically the enterotoxin molecule itself, since elution profiles on Sephadex G-75 chromatography were unchanged after alkylation. Can J Microbiol, 1975 Aug, 21(8), 1144 - 50 Biochemical changes during sporulation of Bacillus stearothermophilus; Orlowski M; The process of sporulation was studied in Bacillus stearothermophilus . A medium is described that supports good growth and sporulation of the organism . In this medium, which contains glucose, salts, and amino acids, acetate starts to accumulate before any of the glucose is catabolized . Enzymes of the tricarboxylic acid cycle are present at all times during growth and sporulation and are found in dormant spores . As the glucose in the culture is consumed, acetate rapidly increases and the pH of the medium drops . The acetate rapidly disappears during sporulation and the pH rises . Dipicolinic acid appears during sporulation and several key-enzyme activities fluctuate in a characteristic pattern. Cancer Res, 1975 Aug, 35(8), 1985 - 90 Observations on trace proteins in plasma of febrile patients by cationic disc electrophoresis in acrylamide gel at pH 3.8; Young CW et al.; Cationic disc electrophoresis at pH 3.5 in 6 M urea-containing acrylamide gels permits analysis of plasma and other body fluids for the presence of trace proteins with pI greater than or equal 5 and M.W . less than 60,000 . These charge and size characteristics would include rabbit and human granulocytic pyrogen, human monocytic pyrogen and, by inference, other similar candidate pyrogenic proteins . Semiquantitation of the trace protein content can be achieved by densitometric scanning of gels stained with Amido schwarz . Duplicate analyses were performed on plasma samples from 133 individuals: normal, 15; afebrile advanced cancer, 18; afebrile Hodgkin's disease, 30; febrile Hodgkin's disease, 33; other febrile lymphoma, 13; febrile advanced cancer without infection, 12; and pyogenic fever, 12 . Plasma from most of the febrile patients, particularly from febrile Hodgkin's disease patients, contained trace proteins not detectable in the afebrile individuals studied . In patients with Hodgkin's disease the quantity of trace proteins present in plasma correlated well with overall severity of Hodgkin's pyrexia, but not with spontaneous hr to hr fluctuations in the fever . Marked reduction in plasma levels of the trace proteins occurred with response to antitumor therapy . Elevated plasma levels of these proteins can be induced by intratumoral inoculation of Bacillus Calmette-Guerin . They appear concomitant with the febrile response. J Cell Biol, 1975 Aug, 66(2), 233 - 42 The relation between sporulation and the induction of antibiotic synthesis and of amino acid uptake in Bacillus brevis; Lee SG et al.; The induction and localization of tyrocidine-synthesizing enzymes is shown to be parallel, during growth of Bacillus brevis (ATCC 8185, American Type Culture Collection, Rockville, Md.), with the induction of uptake of constitutive amino acids and of components of pantetheine, a coenzyme of tyrocidine synthesis . Antibiotic synthesis appears at the end of logarithmic growth when the first soluble enzymes may be obtained from homogenates . During this period, binding proteins for metabolite uptake were isolated by intensive sonication which, when studied by chromatography, were identified by the appearance of low molecular weight fractions binding the radioactively marked metabolites; their induction was prevented by addition of rifampicin . The major purpose of this study was a comparison of antibiotic production and sporulation, the progress of which was followed by electron microscopy . The onset of tyrocidine synthesis and metabolite uptake coincided with the appearance of septum formation indicating that sporulation had progressed to stage II . With the progress of spore encapsulation, the tyrocidine production migrated from the soluble fraction into the forespore, terminating with the separation of forespores from the sporangium membrane . The resulting concentration of antibiotic in the forespore may indicate its function in sporulation, the nature of which, however, was not explored. J Biochem (Tokyo), 1975 Aug, 78(2), 253 - 9 Purification of several bacteriolytic enzymes by affinity chromatography on lysozyme-lysate of Micrococcus lysodeikticus cell wall coupled with sepharose; Yoshimoto T et al.; Using lysozyme-lysate of Micrococcus lysodeikticus cell wall coupled with Sepharose, several bacteriolytic enzymes were purified from crude preparations of animal and microbial origin . Quail egg-white, human milk and salivary lysozymes {EC 3.2.1.17} were adsorbed onto the adsorbent at pH 5-7 and eluted with 2M NaCl at pH 10 . By means of these treatments, lysozymes were purified 20-250 fold with activity recoveries of 60-80%, and the quail lysozyme thus purified was shown to be discelectrophoretically homogeneous . Some bacteriolytic enzymes of microbial origin were also highly purified by using this affinity adsorbent . A bacterial lysozyme from Bacillus sp . ML-208 showed high affinity for the ligand and was not eluted under the conditions mentioned above, but was recovered by elution with 2M guanidine-HCl at pH 5.8, resulting in a 500-fold increase in the specific activity . A Pseudomonas-lytic enzyme from Streptomyces sp . P-51 was easily released from the adsorbent by elution with 0.5M NaCl at pH 5.0 . A staphylolytic F2 enzyme from S . griseus S-35 and a chitinase {EC 3.2.1.14} from yam, both of which were completely inert toward M . lysodeikticus cell wall, passed through the adsorbent column . A modified ligand, in which muramic acid and glucosamine residues were N,O-acetylated, failed to adsorb any of these animal and bacterial lysozymes . Some of the enzymatic properties and bacteriolytic action spectra of these purified enzymes are also described in this paper in comparison with those of hen egg-white lysozyme. Biochemistry, 1975 Jul 29, 14(15), 3344 - 50 Tyrosyl-tRNA synthetase from Escherichia coli . Stoichiometry of ligand binding and half-of-the-sites reactivity in aminoacylation; Jakes R et al.; The tyrosyl-tRNA synthetase from Escherichia coli binds only 1 mol of tRNA, tyrosine, and tyrosyl adenylate per mol of enzyme dimer . However, like the enzyme from Bacillus stearothermophilus, once one active site is occupied by tyrosyl adenylate the other becomes accessible to bind a further molecule each of tyrosine and ATP . Both bacterial enzymes show biphasic kinetics with respect to tyrosine in the aminoacylation of tRNA . Equilibrium dialysis experiments show that this is due to 2 mol of tyrosine binding in the presence of ATP and tRNA . A method is given for a correction for the effects of hydrolysis of the charged tRNA on the aminoacylation kinetics. C R Acad Sci Hebd Seances Acad Sci D, 1975 Jul 28, 281(4), 313 - 6 {Inhibition of cytotoxic effects of B1 aflatoxin towards the bacterial cell by coumarin . Role of other interfering factors}; Boutibonnes P et al.; The coumarin has with some limits a repressive effect on the antibacterial power of Aflatowin B1 . It behaves towards cells of Bacillus thuringiensis (Berliner) treated by the myocotoxic, like a curative agent . Therefore, the reaction of microbial cells on the cytotoxic effect of Aflatoxin B1 depends on the aeration of the culture and chiefly on the "inoculum effect". Biochim Biophys Acta, 1975 Jul 27, 397(1), 188 - 93 Purification and some properties of alkaline pullulanase from a strain of bacillus no . 202-1, an alkalophilic microorganism; Nakamura N et al.; Pullulanase (pullulan 6-glucanohydrolase EC 3.2.1.41) was purified about 290-fold from the culture fluid of Bacillus No . 202-1 by DEAE-cellulose adsorption, acetone fractionation, (NH4) 2SO4 precipitation and DEAE--cellulose column chromatography followed by Sephadex G-200 molecular sieve chromatography . The enzyme gave a single band of protein by disc polyacrylamide gel electrophoresis . The molecular weight was estimated as 92 000 by sodium dodecyl sulfate gel electrophoresis . The isolectric point was lower than pH 2.5 . The optimum pH for enzyme action was about 8.5-9.0 . The action of the enzyme on amylopectin and glycogen resulted in increase in the iodine coloration of 85% and 70%, respectively . The enzyme completely hydrolyzed 1,6-alpha-glucosidic linkages in amylopectin, glycogen and pullulan. Can Med Assoc J, 1975 Jul 26, 113(2), 127 - 8 Interpretation of the PPD skin test in BCG-vaccinated children; Joncas JH et al.; Skin testing with 5 tuberculin units (TU) of purified protein derivative (PPD) of tuberculin stabilized with polysorbate (Tween) 80 was done 3 months and 1 year after immunization with bacille Calmette-Guerin (BCG) vaccine in two groups of children: one group vaccinated at birth and another group at age 6 years . Interpretation of the PPD skin test with 5 TU is possible in children 1 year and older vaccinated with BCG at birth: if the diameter of induration is more than 10 to 12 mm the reaction cannot be ascribed to BCG vaccination and is highly suggestive of supervening infection with Mycobacterium tuberculosis or occasionally atypical mycobacteria . In contrast, the interpretation of a PPD test in children vaccinated at age 6 years is extremely difficult. Biochim Biophys Acta, 1975 Jul 14, 399(1), 31 - 41 Biosynthesis of edeine: II . Localization of edeine synthetase within Bacillus brevis Vm4; Kurylo-Borowska Z; Edeine-synthesizing polyenzymes, associated with a complex of sytoplasmic membrane and DNA, were obtained from gently lysed cells of Bacillus brevis Vm4 . The polyenzymes-membrane-DNA complex, isolated from dells intensively synthesizing edeines (18--20 h culture) contained edeine B . Edeine B was found to be bound covalently t o the edeine synthetase . The amount of edeine bound to polyenzymes was 0.1--0.3 mumol/mg protein, depending on the age of cells . Detachment of deeine synthetase with a covalently bound edeine B from the membrane-DNA complex was accomplished by a treatment with (NH4)2-SO4 at 45--55% saturation or by DEAE-cellulose column fractionation . In contrast to other components of the complex, the edeine-polyenzymes fragment was not adsorbed to the DEAE-cellulose . Sephadex G-200 column chromatography separated the edeine-polyenzymes complex into 3 fractions . Edeine-polyenzymes complex, obtained from lysozyme-Brij-58-DNAase treated cells, contained edeine B bound to two protein fractions of mol . wt 210 000 and 160 000 . Edeine-polyenzymes complex detached from the complex with the membrane and DNA contained edeine B, bound only to protein fraction of mol . wt 210 000 . Edeine A was not found in the edeine-polyenzymes complex . No accumulation of free antibiotics within 16--22 h old cells of B . brevis Vm4 was detected . The edeine-polyenzymes complex associated with the DNA-membrane complex has shown no antimicrobial activity . By treating of above with alkali, edeine B of specific activity: 80 units/mjmol was released . The complex of DNA-membrane associated with edeine-polyenzymes complex was able to synthesize DNA, under the conditions described for synthesis, directed by a DNA-membrane complex . Edeine when associated with this complex did not effect the DNA-synthesizing activity. Appl Microbiol, 1975 Jul, 30(1), 20 - 5 Physiology of sporeforming bacteria associated with insects: metabolism of Bacillus popilliae grown in third-instar Popillia japonica Newman larvae; St Julian G et al.; The timing and relative participation of concurrent pathways of carbohydrate metabolism as well as the extent of terminal respiratory activity were determined by radiorespirometry with 14-C substrates and by enzyme assays for vegetative and sporulating cells of the bacterium Bacillus popilliae cultured in whole, intact Popillia japonica (Japanese beetle) larvae . During vegetative proliferation, the pentose phosphate pathway predominates in the bacterial cells with minor involvement of the Embden-Meyerhof-Parnas pathway . As the cells proceed through sporulation, pentose phosphate and Embden-Meyerhof-Parnas activity remains constant . No tricarboxylic cycle activity is evident during growth and sporulation of B . popilliae . The results demonstrate (i) predominantly aerobic metabolism for carbohydrate assimilation within in vivo sporulating cells, (ii) a major contrast to the metabolism of other aerobic sporeforming bacteria that exhibit derepression of tricarboxylic acid cycle enzymatic activity at the onset of sporulation, and (iii) no causal necessity of the cycle to B . popilliae sporogeny. Biochim Biophys Acta, 1975 Jul 7, 395(3), 373 - 80 Association of ribosomal subunits . IV . Polyamines as active components of the association factor from Bacillus stearothermophilus; Garcia-Patrone M et al.; The association of ribosomal subparticles induced by several associating agents has been studied under different conditions . The following observations were made: 1 . Spermidine was able to produce the association of subunits, and the concentration and temperature curves of this reaction were similar to those obtained with association factor . The product formed in the latter case was more stable . 2 . The association at low Mg2+ concentration was higher with association factor than with polyamines . 3 . The temperature-dependent binding of spermidine to 30-S subunits formed an active complex, which was converted into the 30S-50S couples by the addition of 50-S subparticles at low temperature . A similar behaviour has been previously shown for the complete association factor and its low molecular weight fraction . 4 . The same unstable form of 30S-50S couples has been obtained either with spermidine or with the low molecular weight component (AFII) of the association factor . In both cases the protein fraction AFI was able to complete the reaction by stabilizing the subunit couple . 5 . After glutaraldehyde fixation the products of the reactions with spermidine or association factor behaved in a similar way when they were submitted to long sucrose-gradient centrifugations . 6 . The analysis of association factor preparations has shown that they contain spermidine as well as spermine . The polyamine levels in association factor could account for part of the total associating activity. J Neurol, 1975 Jul 2, 209(3), 181 - 7 {Bacillary dysentery with involvement of the central and peripheral nervous system (author's transl)}; Pilz P et al.; A man, age 50, fell ill with polyneuropathy followed by parkinsonism and organic psychosis and died in a shock 6 weeks later . Serologic examination suggested bacillary dysentery, but the patient had no diarrhoea . The neuropathological examination did not reveal any organic substrat of parkinsonism . Peripheral nerves showed mucoid degeneration, segmental demyelination and lymphocytic infiltration of peri- and endoneurium . Many Renaut bodies were found which seemed to arise from mucoid masses organized by cells of the endoneurium . Polyneuropathy and parkinsonism are well known neurological complications of bacillary dysentery and favour this diagnosis in accord with the serological findings. Bull Soc Pathol Exot Filiales, 1975 Jul-Aug, 68(4), 367 - 70 {Demonstration of Pseudomonas pseudomallei (Whitmore's bacillus) in the mud of Iranian ricefields (author's transl)}; Pourtaghva M et al.; Whitmore bacillus had killed a horse and a mule in a herd bred for serum production in 1970 (Baharsefat and Amjadi) . Out of 157 soil samples from rice fields in Northern Iran it was possible to isolate 19 times Pseudomonas pseudomallei (types I and II) possessing a very high pathogenicity for animals . Attempts at an evaluation of the human incidence of the disease. Mol Biol (Mosk), 1975 Jul-Aug, 9(4), 602 - 8 {Some properties of 1 P+f bacteriophage for Bacillus brevis var . G.-B and its nucleic acid}; Dobritsa AP et al.; The 1 P+f phage, a virulent mutant of the moderate P+ phage for Bac . brevis var . G.-B., consists of a hexagonal head (90x90 nm) and a long non-contractile tail (340 nm) . This phage is characterized by a relatively long latent period (90-110 min) and a low yield (40-50 particles per cell) . The 1P+f phage is quite stable at pH values from 1 to 11, insensitive to osmotic shock, treatment with chloroform and acridine orange . The sensitivity of the phage to thermal treatment and UV-radiation has been studied . The nucleic acid of the P+f phage is double-stranded DNA of AT-type (GC equals 34.5 mole %) which contains 5-methylcytosine (0.18 mole %) and N6-methyladenine (0.32 mole%) . The level of methylation of cytosine and adenine residues in DNA of the 1 P+f phage does not depend on the host studied (Bac . brevis, P- and S variants) . The specificity of methylation of cytosine residues in the S and P- cells appears to be the same . DNA of the 1 P+f phage strongly differs from DNA of the host in nucleotide composition (GC equals 45.7 mole %) . Nevertheless, phage DNA is very similar to DNA from Bac . subtilis in the character of pyrimidine distribution (the amount of different pyrimidine isopliths) . This may testify to a somewhat common character of the nucleotide sequence organization in DNA of the phage and its host. Prikl Biokhim Mikrobiol, 1975 Jul-Aug, 11(4), 550 - 5 {Study of catalase and proteolytic activities of different variants of Bacillus mesentericus}; Vasilevskaya IA et al.; Catalase and proteolytic activity of the culures and morphological variants of Bacillus mesentericus fuscus, Bac . mesentericus vulgatus were studied . The variants were obtained as a result of prolonged cultivation of the stock strains in the potato mash under the layer of vaseline oil . The level of catalase activity varies in different morphological variants of the same culture, changes with age and depends on the storage conditions . The catalase activity in the rough, smooth and papillar variants that were freshly isolated from the potato mash was 1.5=2.5 times lower than that in the variants long kept on the agar medium . The quantitative indexes of the proteolytic activity of different variants also varied. J Biochem (Tokyo), 1975 Jul, 78(1), 115 - 21 A nucleoside triphosphate-dependent deoxyribonuclease from Bacillus laterosporus . The mode of action of the enzyme; Anai M et al.; The acid-soluble products of exhaustive digestion of native DNA with Bacillus laterosporus DNase consist of 6.5% of mononucleotides and 93.5% of oligonucleotides with an average chain length of 3.2 . The results of viscometric studies and inactivation of transforming DNA indicate the existence of acid-insoluble intermediates and the selective degradation of the population of substrate molecules rather than a random nucleolytic action . Furthermore, sucrose density gradient analysis of partially digested DNA showed that the initial DNA added as a substrate disappeared progressively during the reaction, being replaced by much more slowly sedimenting acid-insoluble materials, which were eventually degraded into acid-soluble end products during the reaction; products intermediate in size between these two components were not detectable . Studies with DNA labeled at the 3'-terminus indicate that Bacillus laterosporus DNase does not attack DNA from 3'-hydroxyl ends to yeild acid-soluble or acid-insoluble materials in a random manner . The results presented in this paper indicate that the nature of the attack of B . laterosporus nuclease is similar to that previously proposed for Micrococcus luteus DNase . The possibility of the sequential release of acid-insoluble intermediate fragments as well as acid-soluble products from the terminal portion of DNA by the enzyme is discussed. J Bacteriol, 1975 Jul, 123(1), 377 - 9 Evidence for a nonrandom base sequence in a Bacillus pumilus plasmid: EcoR1 endonuclease digestion of pPL576; Lovett PS et al.; EcoR1 endonuclease digested the Bacillus pumilus plasmid pPL576 (molecular weight similar to 28 X 10-6) into three distinct size classes of linear fragments . The molecular weights of the fragments are 13.0 X 10-6, 0.5 X 10-6, and 6.5 X 10-6 by sucrose gradient analysis . By electron microscope analysis the three fragments account for about 99% of the intact plasmid . These results indicate that pPL576 molecules contain a nonrandom base sequence, and are consistent with the interpretation that pPL576 is autonomous and not the result of cyclization of random chromosome fragments. Surgery, 1975 Jul, 78(1), 66 - 75 Effect of bacillus Calmette-Guerin immunotherapy on tumor antigen-induced lymphocyte-stimulated protein synthesis in melanoma patients; Roth JA et al.; Changes in in vitro lymphocyte-stimulation protein synthesis (SPS) of 40 melanoma patients following incubation with 3M KCl extracts of allogenic melanoma, lung carcinoma, and sarcoma antigens and phytohemagglutinin (PHA) were quantitated by measuring H-3-leucine uptake . One of eleven "untreated" melanoma patients stimulated significantly to the melanoma antigen . However, this lymphocyte response was not significantly different from that of the normal subjects . Patients who received systemic bacillus Calmette-Guerin (BCG) by the tine technique for 3 months and for 6 months had significant increase in lymphocyte protein synthesis following incubation with melanoma antigen . There were no significant differences in PHA responses between the "untreated" melanoma patients and the BCG-treated group . Testing of serial lymphocyte samples from nine melanoma patients before treatment and at monthly intervals thereafter confirmed these observations . Furthermore, no change in serial complement-fixing antibody titers to melanoma antigen was noted in the BCG-treated patients . These results demonstrated that in vitro lymphocyte responses to melanoma antigen may be augmented by BCG therapy. Cancer Res, 1975 Jul, 35(7), 1779 - 90 Effects of methanol extraction residue and therapeutic irradiation against established isografts and simulated local recurrence of mammary carcinomas; Yron I et al.; Female BALB/c mice carrying established isografts or simulated local recurrence implants of 2 rapidly growing mammary adenocarcinomas were treated either by injection of the methanol extraction residue (MER) fraction of killed Bacillus Calmette-Guerin organisms (given s.c . or into the tumor) or by focal X-irradiation or by both . None of the modalities of therapy effected cures, but in many instances there was a significant retardation of tumor cevelopment and prolongation of the lives of the mice . Administration of MER alone offered protection in a number of cases but less often than the other forms of treatment . Combined therapy with MER and irradiation was, on the whole, the most successful therapeutic intervention . MER or irradiation administered alone enhanced the neoplastic process only on rare occasions; this appeared to be the case even more infrequently with combined treatment . MER was most likely to be effective alone or in combination when small quantities were used and when only 1 treatment or 1 cycle of combined therapy was given . The therapeutic action of MER was not dependent on direct introduction of the agent into a neoplastic focus; s.c . administration distal to the tumor site was almost always at least as satisfactory as injection directly into the tumor mass and indeed was often more efficacious. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jul, (7), 68 - 72 {Current data on the polymorphism of Rickettsia prowazekii and burneti in cultured cells}; Gulevskaia SA et al.; In cultivation of Rickettsia prowazeki (strains Breinl and E) in the cell cultures of guinea pig kidneys (GPK) and chick embryo fibroblasts (CEF) ultrastructure of rickettsia of unusual shape (filamentous, irregularpleomorphic and spheroplast-like) were revealed along with rickettsia of the usual shape and size . The polymorphism was less pronounced in the GPK and the CEF cells of Rickettsia burneti (strain M-44) . It is supposed that rickettsial polymorphism was not associated with their developmental cycle and served as a morphological expression of the changes in the microorganism under the effect of unfavourable ecological conditions . The appearance of filamentous forms could be associated with disturbed cell division process; changed rigidity of the cell wall could serve as the cause of appearance of pleomorphic rickettsia . In difference from polymorphism, the cycle of rickettsial development is considered to be (in the basis of modern electron microscopic data) as a biological replacement of the vegetative (rod-like, bacillary) forms by those more stable in the external environment, resting (coccoid). Mikrobiologiia, 1975 Jul-Aug, 44(4), 720 - 6 {Bacillus megaterium as a possible source of protein}; Zalabak V et al.; The amino acid composition of protein was studied with Bacillus megaterium . Most of the essential amino acids are present in sufficient amounts, with the exception of sulphur-containing amino acids and probably tryptophan . The content of methionine is 2 percent . The proteins of the biomass are easily digested by trypsin . The culture of Bac . megaterium grows in a mineral medium containing 5 percent of sucrose or glucose . The yield of dry biomass recalculated for the sugar consumed decreases from 0.6 to 0.3 with an increase of the sugar concentration . The addition of molasses or corn steep increases the yield of the biomass to 15--20 g of dry matter per one litre of the medium . The biomass grown under these conditions contains 8--9 percent of nitrogen, 40 percent of protein, up to 30 percent of poly-beta-hydroxybutyrate in the form of granular inclusions, and 10--14 percent of RNA. Am Rev Respir Dis, 1975 Jul, 112(1), 43 - 7 Tuberculin-induced lymphocyte transformation and skin reactivity in monkeys vaccinated or not vaccinated with Bacille Calmette-Guérin, then challenged with virulent Mycobacterium tuberculosis; Chaparas SD et al.; The in vitro lymphocyte transformation test was compared to the skin test at intervals after aerogenic administration of bacille Calmette-Guerin vaccine to monkeys, and also at monthly intervals after aerogenic challenge of monkeys vaccinated and not vaccinated with virulent strain H37Rv of Mycobacterium tuberculosis . Individual responses varied, but several consistent patterns of sensitivity development could be discerned . The lymphocyte transformation test was more sensitive, and often positive when the skin test was negative, doubtful, or feeble . Conversion to tuberculin reactivity was detected by lymphocyte transformation in vitro earlier in the disease or after vaccination, and persisted longer after sensitivity to the skin test had waned or after the animals had become anergic by the skin test . Monkeys not vaccinated, but challenged, developed larger in vitro skin reactions and responses than animals that were either vaccinated and challenged or only vaccinated; however, the unvaccinated, challenged monkeys developed anergy to tuberculin and progressive disease more rapidly than other groups, and their cells became less responsive to phytohemagglutinin in vitro. J Bacteriol, 1975 Jul, 123(1), 354 - 65 Properties of Bacillus cereus spore coat mutants; Aronson AI et al.; Two classes of spore mutants have been selected in Bacillus cereus T, those producing lysozyme-sensitive spores, and those producing spores dependent upon lysozyme for germination . One mutant from each class was studied in detail and found to have defective packing of the spore coat layers . The major spore coat poplypeptide appeared to be altered on the basis of gel electrophoretic profiles and/or peptide maps of half-syctine-containing peptides . The spores of the mutants of both classes were sensitive to lysozyme and failed to respond to the germinants L-alanine plus adenosine . The spores were also more sensitive to octanol than the parental strain, but contained the same amount of dipicolinic acid and were equally heat resistant . The reversion frequencies in both cases were consistent with an initial point mutation, suggesting that an alteration in the major coat polypeptide accounted for the phenotypic properties studied. Mikrobiologiia, 1975 Jul-Aug, 44(4), 645 - 50 {Effect of hydrogen ions on the morphology and ultrastructure of Bacillus megaterium}; Poglazova MN et al.; The morphology and ultrastructure of the cells of Bacillus megaterium depend on the concentration of hydrogen ions in the medium which affect, first of all, the function of division . The changes induced by hydrogen ions are temporary and disappear when the cells are put under normal conditions of growth . The "acid-resistant" strain of Bac . megaterium remains viable at a very high concentration of hydrogen ions (pH=4.2) and, though its morphology changes drastically, no essential damages are found, i.e . the cells become adapted to new conditions . The changes are reversible. Ann Surg, 1975 Jul, 182(1), 15 - 21 Wound infection during the Yom Kippur war: observations concerning antibiotic prophylaxis and therapy; Klein RS et al.; Eighty-eight episodes of wound associated infection were identified among 624 consecutively admitted battlefield casualties . Ninety-one per cent of infections occurred during the administration of antibiotic therapy or prophylaxis and 65% were associated with the use of multiple antibacterial agents . Gram negative bacillary and mixed microbial infection predominated and were found to increase in relative incidence after the second day of hospitalization . Appropriate therapy, based on disc sensitivity testing, was administered in only 33% of infectious episodes . The practice of antibiotic wound prophylaxis may contribute to the incidence and nature of infection in battlefield wounds . Problems unique to the handling of battlefield wounded are discussed in comparing the present data with those of other war associated and civilian studies. J Gen Microbiol, 1975 Jul, 89(1), 93 - 101 The location of bacterial antigens on sections of Bacillus cereus by use of the soluble peroxidase--anti-peroxidase complex and unlabelled antibody; Short JA et al.; The location of antigens on sections of bacteria using the soluble peroxidase-anti-peroxidase complex in conjunction with unlabelled antibody is described . Using this technique, spore antigens have been detected in the cytoplasm of vegetative cells during forespore septum formation and subsequent stages of sporulation . Antigenic sites were first associated with poly-beta-hydroxybutyric acid granules and subsequently were found in increasing quantities in the cytoplasm of the sporangium . Vegetative cell antigens were located on the cell wall and in the cortical region during sporulation . During germination antigens were located in the cortical region, and during outgrowth on the cell wall . These findings are discussed in the light of existing biochemical data. Parazitologiia, 1975 Jul-Aug, 9(4), 373 - 6 {2 new species of microsporidians (Protozoa, Microsporidia) from mosquitoes of the family Chironomidae}; Voronin VN; Two new species of microsporidians are described from the mosquitoes of the genus Chironomus collected in water bodies of the north-eastern USSR . Bacillidium chironomi sp . n . injuring the adipose tissue of the fourth stage larvae of Chironomus dorsalis Mg . has rod-shaped spores 15 (11 to 19) x 0.7 (0.6 to 0.9) mu in size . Duboscqia chironomi sp . n . injures the adipose tissue of the fourth stage larvae of Chironomus plumosus L . Fresh spores of this species are egg-shaped, 6.2 (5.8 to 6.5) x 3.8 (3.6 to 4.0) mu in size . When stained according to the Romanovsky-Gimza method the spores are 5.4 (4.9 to 5.8) x 3.8 (3.6 to 4.1) mu in size . Some peculiarities of the nuclei fission in the sporants of Duboscqia chironomi sp . n., are discussed. J Biochem (Tokyo), 1975 Jul, 78(1), 105 - 14 A nucleoside triphosphate-dependent deoxyribonuclease from Bacillus laterosporus . Purification and characterization of the enzyme; Anai M et al.; A deoxyribonuclease, which requires nucleoside triphosphate for reaction, has been purified about 150-fold from extracts of Bacillus laterosporus . Potassium phosphate and ethylene glycol stabilize the purified enzyme . The enzyme degrades double-stranded DNA about 100 times faster than heat-denatured DNA in the presence of nucleoside triphosphate . Double-stranded DNA is not degraded to any measurable extent in the absence of ATP, but the enzyme exhibits activity toward denatured DNA in the absence of nucleoside triphosphate, and this activity seems to be an intrinsic property of this enzyme protein . The optimum pH is 8.5 and the maximum activity is obtained in the copresence of Mg2+ (8.0 X 10(-3)M) and Mn2+ (7.0 X 10(-5)M) . ATP and dATP are most effective and nucleoside di- or monophosphates are ineffective . ATP is converted to ADP and inorganic phosphate during the reaction and the ratio of the amount of ATP cleaved to that of hydrolyzed phosphodiester bonds of DNA is about 3:1 . An inhibitor of the enzyme was observed in bacterial extracts prepared by sonic disruption; the inhibitory substance is produced in the bacteria in the later stages of cell growth . Preliminary results show that the inhibitor emerged near the void volume of a Sephadex G-200 column, and was relatively heat-stable, RNase-resistant, and DNase-sensitive. Boll Ist Sieroter Milan, 1975 Jun 26, 54(2), 98 - 104 Study of the sporulation of Bacillus thuringiensis var . thuringiensis; Rendic S et al.; During the submerged cultivation of Bacillus thuringiensis var . thuringiensis in 300- and 3000-liter fermentors, lysis occurred at the end of the exponential phase of growth . New vegetative cells were subsequently formed which usually sporulated . At time of lysis, the amount of soluble sugar was 1-12 g/liter, pH value dropped to 5.3-5.8 from the original PH 6.8 and started to rise after all the cells had lysed . The proteolytic activity was low during the lysis and increased as the sporulation commenced. Biochim Biophys Acta, 1975 Jun 24, 391(2), 435 - 47 Bacillus cereus beta-lactamase . Reaction with N-bromosuccinimide and the properties of the product; Ogawara H et al.; The effect of N-bromosuccinimide on the enzymatic activity and the conformation of a Bacillus cereus beta-lactamase (penicillin amido-beta-lactamase EC 3.5.2.6) was studied . Incubation with 10 muM N-bromosuccinimide caused over 95% decrease of the enzymatic activity within 15 min . Spectrophotometric titration with N-bromosuccinimide showed that the reaction proceeded in two steps . The half-inactivated enzyme was prepared by the reaction with N-bromosuccinimide and its properties examined . Amino acid analysis showed that the half-inactivated enzyme contained one residue of tryptophan less while other amino acid contents were similar . Neither the molecular weight nor the mobility in disc electrophoresis was changed . However, the affinity to a cephalexin-CH-Sepharose column was increased, and the Km value for cloxacillin was one-third that of the native enzyme, although that for benzylpenicillin was similar . These results indicate that a tryptophan residue sensitive to N-bromosuccinimide is essential for the maintenance of the rigid conformation and that its oxidation alters the enzyme in a manner such that a substrate with a bulky group in its side chain can form an enzyme-substrate complex more easily . In the native enzyme, the value of (f(a))(eff) (Lehrer, S.S . (1971) Biochemistry 10, 3254-3263), did not vary significantly in the absence or the presence of cloxacillin . In contrast, in the half-inactivated enzyme the presence of cloxacillin affected the conformation such that over two thirds of the tryptophyl fluorescence were accessible for quenching by KI, although about half was accessible in the absence of cloxacillin. Biochim Biophys Acta, 1975 Jun 24, 391(2), 326 - 33 The metal ion dependence of phospholipase C from Bacillus cereus; Little C et al.; 1 . The zinc content and metal ion dependence of phospholipase C(phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) from Bacillus cereus have been examined . 2 . The native enzyme contained about 2 atoms of tightly bound zinc/molecule . 3 . Incubation of the enzyme with EDTA or with o-phenanthroline caused inactivation . The inactivation was accompanied by the removal of one zinc atom from the enzyme and could be fully reversed by the addition of Zn2+ or Co2+ to the enzyme and partly reversed by Mn2+ or Mg2+ . 4 . Prolonged exposure to o-phenanthroline removed the second zinc atom also and produced an enzyme species which was reactivated by Zn2+ only . Full reactivation was accompanied by the binding of about two zinc atoms to the enzyme . 5 . The results are consistent with the view that phospholipase C is a zinc metalloenzyme. C R Acad Sci Hebd Seances Acad Sci D, 1975 Jun 23, 280(24), 2801 - 4 {Chemically controlled depolymerization of beta hydroxybutyric lipids (PHB) in Bacillus megaterium . Isolation and structure of the oligomers of D(-)beta hydroxybutyric acid}; Hauttecoeur B et al.; Partial depolymerisation of PHB by chemical means to the appearance of homologous polymers from PHB constituted of short carbon chains and oligomers which represent the first elements of this macromolecule . The chemical structure of these oligomers ranging from dimer to heptamer has been essentially deduced from their mass spectrum and then confirmed by studying their physical, chemical and biological properties. Biochim Biophys Acta, 1975 Jun 23, 388(3), 305 - 17 Omega-1, Omega-2 and Omega-3 hydroxylation of long-chain fatty acids, amides and alcohols by a soluble enzyme system from Bacillus megaterium; Miura Y et al.; A soluble enzyme preparation from Bacillus megaterium, previously shown to hydroxylate free fatty acids to isomeric mixtures of Omega-1, Omega-2 and Omega-3 monohydroxy fatty acids in the presence of NADPH and O2, has now been shown to act also on fatty amides but not only hydrocarbons or fatty acid methyl esters . Using 14-C-labelled substrates, both the chain-length specificity and the positional specificity of hydroxylation was determined for fatty acids, alcohols and amides . The most active saturated fatty acid (pentadecanoic) was hydroxylated at a rate 10 times greater than the most active amide (myristamide) and 14 times faster than the most active alcohol (1-tetradecanol) . Among the saturated fatty acids, the order of activity as hydroxylation substrates was C15 greater than C16 greater than C14 greater than C17 greater than C13 greater than C18 = C12 . For amides the order was C14 greater than C12 greater than C15 greater than C16 while for alcohols it was C14 greater than C13 = C15 greater than C12 greater than C15 . Four cis-monounsaturated fatty acids were also tested . Oleic, palmitoleic and cis-12-octadecenoic acids were more active than their saturated analogs but cis-5-tetradecenoate was less active than myristate . For all of the substrates mentioned above, with the possible exception of several unsaturated acids, the alkyl chains were monohydroxylated to give isomeric mixtures of the Omega-1, Omega-2 and Omega-3 derivatives . The distribution of these three isomers varied with chain-length and type of substrate but generally, the Omega-2 position was favored . The terminal methyl (Omega) group of these substrates was never hydroxylated and there did not appear to be significant hydroxylation of methylene carbons beyond the Omega-3 position . Based on the data presented here and in a previous paper, a model is proposed for the enzyme-substrate complex which involves hydrophobic binding and sequestering of the terminal methyl group of the substrate and electrostatic binding of the substrate's polar functional group. Eur J Biochem, 1975 Jun 16, 55(1), 131 - 9 Morphogenesis of the membrane-bound electron-transport system in sporulating Bacillus megaterium KM; Wilkinson BJ et al.; The properties of electron transport systems present in soluble and particulate fractions of spores of Bacillus megaterium KM?HAVE BEEN COMPARED WIth those of similar fractions prepared from exponential-phase vegetative cells of this organism . The timing and localization of modifications of the electron transport system occurring during sporulation have been investigated by using a system for separating forespores from mother cells at all stages during development {8} . Spore membranes contained cytochromes a + a3, and o at lower concentrations than in vegetative membranes, and in addition cytochrome c, which was not found in exponential-phase vegetative membranes . An NADH oxidase activity of similar specific activity was found in both spore and vegetative membranes but DL-glycerol 3-phosphate and L-malate oxidase activities were found only in vegetative membranes . A soluble NADH oxidase of low specific activity was found in spores and vegetative cells which probably involves a flavoprotein reaction with oxygen because the activity was stimulated by FAD or FMN and difference spectra of concentrated soluble fractions showed spectra typical of a flavoprotein . Particulate NADH oxidase was sensitive to all classical inhibitors of electron transport tested whereas soluble NADH oxidase was insensitive to many of these inhibitors . Cytochrome c was formed between stage I and II of sporulation and this coincided with a five-fold increase in NADH-cytochrome c reductase activity . Forespore membranes had lower contents of cytochromes than sporangial cell membranes but similar levels of NADH and L-malate oxidases; DL-glycerol 3-phosphate oxidase activity could not be detected in either membranes by stage III of sporulation . This characterization of spore electron transport systems provides a basis for suggestions concerning initial metabolic events during spore germination and the effect of a number of germination inhibitors. Int J Cancer, 1975 Jun 15, 15(6), 897 - 911 Tumour inhibition mediated by BCG in immunosuppressed rats; Moore M et al.; Two rat sarcomas (CC5 and P7) which grow progressively on transplantation into normal syngeneic hosts failed to develop when injected in admixture with the Glaxo strain of Bacillus Calmette-Guerin (BCG) . Under comparable conditions, the local development of a third neoplasm (P8) was temporarily inhibited and the number of pulmonary metastases significantly reduced . Experiments were undertaken to determine the extent to which the anti-tumor action of BCG required an immunocompetent host . Rats were immunosuppressed by sub-lethal whole-body irradiation (450 R), with or without prior thymectomy and challenged with inocula of mixed BCG and tumour cells when their capacity to respond to bacterial, tumour and unrelated antigens was maximally depressed . In non-sensitized immunosuppressed rats, the ability of BCG to limit tumour outgrowth was abrogated only in the case of sarcoma CC5 . For this neoplasm, immunogenic in syngeneic hosts by conventional criteria, there was a statistically significant difference in the number of tumours in immunosuppressed rats (51%) compared with non-sensitized immunocompetent controls (6%) . Presensitization to either bacterial or tumour antigens, prior to thymectomy and/or irradiation, fully restored the tumour-inhibitory capacity of BCG . By contrast, sarcoma P7 was not significantly less susceptible to BCG-induced regression in non-sensitized immunosuppressed rats than in nonsensitized normal rats; and sarcoma P8 similarly failed to reveal any significant differences in susceptibility to BCG affecting primary or secondary tumour development . It is concluded that tumours may vary widely in their sensitivity to host reactions aroused by BCG . Certain neoplasms, exemplified by sarcoma CC5, require participation of an immune reaction of delayed hypersensitivity type for optimal destruction at BCG sites, while for others (e.g . sarcoma P7) an immunoreactive component of this type is not essential . By contrast, a third category of tumour (e.g . sarcoma P8) is relatively resistant to host reactions induced by the mycobacteria . An important component of BCG-mediated tumour inhibition is not dependent on an immunologically intact host. Biochemistry, 1975 Jun 3, 14(11), 2367 - 70 A two-state conformational transition of the extracellular ribonuclease of Bacillus amyloliquefaciens (barnase) induced by sodium dodecyl sulfate; Hartley RW; Barnase, the extracellular ribonuclease of Bacillus amyloliquefaciens, is shown to undergo a reversible two-state conformational transition at 0.65 mM sodium dodecyl sulfate (SDS) AAT 37 DEGREES . The prinicipal evidence is based on the equivalence of two independent values of the SDS-barnase binding ratio; about 14 mol of SDS/mol of barnase . Both were derived from fluorometric titration data, one being based on simple conservation of SDS and the other on the use of Wyman's theory of linked functions . No SDS is bound to barnase at SDS concentrations below the transition region. Biochem J, 1975 Jun, 148(3), 513 - 20 Enzymological aspects of the pathways for trimethylamine oxidation and C1 assimilation of obligate methylotrophs and restricted facultative methylotrophs; Colby J et al.; Extracts of trimethylamine-grown W6A and W3A1 (type M restricted facultative methylotrophs) contain trimethylamine dehydrogenase whereas similar extracts of Bacillus PM6 and Bacillus S2A1 (type L restricted facultative methylotrophs) contain trimethylamine mono-oxygenase and trimethylamine N-oxide demethylase but no trimethylamine dehydrogenase . Extracts of the restricted facultatives and of the obligate methylotroph C2A1 contain hexulose phosphate synthase-hexulose phosphate isomerase activity; hydroxypyruvate reductase was not detected . Neither the restricted facultatives nor the obligates 4B6 and C2A1 contain all the enzymes of the hexulose phosphate cycle of formaldehyde assimilation as originally proposed by Kemp & Quayle (1967) . Organisms PM6 and S2A1 lack transaldolase and use a modified cycle involving sedoheptulose 1,7-diphosphate and sedoheptulose diphosphatase . The obligates 4B6 and C2A1, and the type M organisms W6A and W3A1, use a different modification of the assimilatory hexulose phosphate cycle involving the Entner-Doudoroff-pathway enzymes phosphogluconate dehydratase and phospho-2-keto-3-deoxygluconate aldolase . The lack of fructose diphosphate aldolase and hexose diphosphatase in these organisms may be a partial explanation of their restricted growth-substrate range . Enzymological evidence suggests that all the obligates and the restricted facultatives use a dissimilatory hexulose phosphate cycle to accomplish the complete oxidation of formaldehyde to CO2 and water. J Gen Virol, 1975 Jun, 27(3), 305 - 12 Electron microscopical observations of the structure of the virus of viral haemorrhagic septicaemia (VHS) of rainbow trout (salmo gairdneri); Olberding KP et al.; Negative staining of particles of the Danish F-1 strain of viral haemorrhagic septicaemia virus grown in rainbow trout gonad-2 cells revealed three different types of particles: bacilliform, bullet-shaped, and long particles . The average size of the first two types was 165 x 65 nm . The long particles had a length of up to 3000 nm and showed a close resemblance to Marburg virus . Well defined surface projections could be found in all particle types . Without any special treatment we were able to demonstrate different disintegration stages of VHS virus particles in all preparations. Jpn J Microbiol, 1975 Jun, 19(3), 187 - 92 Changes of ultrastructure in spore coat of Bacillus thiaminolyticus during germination and outgrowth; Watabe K et al.; Electron microscopic observation showed that the spore coat of Bacillus thiaminolyticus consisted of at least four layers; a high electron dense outer spore coat layer with five prominent ridges, a middle spore coat layer including two layers of a high and a low electron density, and an inner spore coat layer composing six to seven laminated layers . Rapid breakdown of the cortex and swelling of the core occurred in spores which were allowed to germinate by L-alanine for 45 min, whereas no change of surface feature was observed by scanning electron microscopy . Germination and outgrowth of spores in nutrient broth proceeded, being accompanied by morphological changes, in three steps; the first is a rapid breakdown of the cortex and swelling of the core, the second degradation of the inner layer at prominent region of the spore coat, and the last rupture of the spore coat and emergence of a young vegetative cell. Appl Microbiol, 1975 Jun, 29(6), 717 - 21 Inhibition of phenylamide hydrolysis by Bacillus sphaericus with methylcarbamate and organophosphorus insecticides; Engelhardt G et al.; The degradation of the phenylamide herbicides monolinuron, linuron, and solan by cultures of Bacillus sphaericus ATCC 12123 was inhibited by the methylcarbamate insecticides metmercapturon, aldicarb, propoxur, and carbaryl and by the organophosphorus insecticides fenthion and parathion . The extent of inhibition was largest with metmercapturon and smallest with parathion inhibition of hydrolysis of the two phenylurea herbicides was greater than of the acylanilide compound . Tests with crude enzyme preparations of aryl acylamidase derived from B . sphaericus showed that the inhibition of the hydrolysis of linuron with methylcarbamates is a competitive one . The insecticides tested did induce the enzyme, nor could they serve as its substrate. Br J Exp Pathol, 1975 Jun, 56(3), 265 - 70 The effect of a tubercle lipid adjuvant on the distribution of injected foreign red blood cells; Donald KJ et al.; Increased localization occured of injected foreign red cells in the spleen and lungs of animals treated with a tubercle bacillary lipid adjuvant given intravenously . The distribution changes varied depending upon the time interval between injections of the adjuvant and the foreign red cells . These changes offer an explanation of the augmentation of haemolysins and haemagglutinins previously shown for the lipid. J Bacteriol, 1975 Jun, 122(3), 1322 - 38 Ultrastructural studies of sporulation in Bacillus sphaericus; Holt SC et al.; Spore septum formation in Bacillus sphaericus 9602 occurs 2 h after the end of exponential growth at one end of the vegetative cell, which retains a uniform diameter . The apparently rigid spore septum contains an inner cell wall layer which disappears when the sporulation septum "bulges" into the mother cell cytoplasm . This process occurs simultaneously with terminal swelling at the end of the cell containing the spore septum . It is suggested that the inner cell wall layer is peptidoglycan and that its dissolution and the terminal swelling are consequences of a localized autolysis . Engulfment of the forespore by membrane proliferation results in the production of a forespore surrounded by two flexible, closely apposed membranes . These membranes appear to become more rigid as a peptidoglycan-like layer appears between them, concomitant with the condensation of the forespore nucleoid into a crescent-shaped structure . After nuclear condensation, visible development of distinct cortex, primordial cell wall, and spore coat layers begin, and the forespore cytoplasm assumes an appearance similar to that of a refractile spore . The spore coats consist of an amorphous inner layer, a lamellar midlayer, and a structured outer layer . As cortex synthesis and spore coat assembly continue, exosporium development commences close to that portion of the mother cell plasma membrane wh