Bacillus

Can Med Assoc J, 1977 Jul 9, 117(1), 33 - 6
Recurrent malignant melanoma: effect of adjuvant immunotherapy on survival; McCulloch PB et al.; Twenty-nine patients referred consecutively to a cancer clinic because of recurrent metastatic malignant melanoma were given 5 mg of Connaught Laboratories bacillus Calmette-Guerin (BCG) by multiple cutaneous puncture at weekly and later at monthly intervals . Eight were also treated with autologous tumour vaccine and three with intralesional BCG . This group was compared with a retrospective control group of 54 patients treated with surgery and radiotherapy alone after recurrence . Prognostic features such as site of primary and of first metastasis, disease-free interval, age and sex were similar in the two groups . However, the median survival from the time of first recurrence was 12 months in the control group but 21 months in the BCG-treated group . The major improvement was in patients with disease limited to the regional lymph nodes: the median survival was 16 months in the control group but over 32 months in the BCG-treated group . Autologous tumour vaccine appeared to have no effect on survival . Serial testing of immunocompetence did not offer any prognostic advantage, although the results of some tests correleated well with extent of disease.

Biochim Biophys Acta, 1977 Jul 7, 461(1), 109 - 23
Cyanide-resistant electron transport in sporulating Bacillus megaterium KM; Hogarth C et al.; The NADH oxidase activity of stage V mother-cell membranes, isolated from sporulating Bacillus megaterium KM, shows a greater inhibition by cyanide and displays this response at lower concentrations of cyanide than the stage V forespore inner membrane . Comparison of the effects of various respiratory inhibitors reveals that the difference in cyanide sensitivity between these membranes is located on the oxidase side of the 2-heptyl-4-hydroxyquinoline N-oxide-sensitive step . Both membranes contain cytochromes a+a3, b-562, b-555, c and d, with three potential oxidases: cytochromes a+a3, o and d . Cyanide difference spectra suggest that cytochromes b-562 and d may be the components involved in the cyanide-resistant electron transport pathway . Membrane ascorbate-N,N,N',N'-tetramethylphenylenediamine and ascorbate 2,6-dichlorophenolindophenol oxidase activities are highly sensitive to cyanide . Evidence is presented for terminal branching of the respiratory chain with branches differing in cyanide sensitivity . The cyanide sensitivity of the NADH oxidase of membranes prepared from various stages of sporulation is compared . Morphogenesis of the mother-cell plasma membrane to a cyanide-sensitive form during stages II and III of sporulation is postulated.

Mikrobiologiia, 1977 Jul-Aug, 46(4), 730 - 6
{Polylysogeny in Bacillus thuringiensis var . Galleriae cultures}; Kochkina ZM et al.; The culture of Bacillus thuringiensis var . galleriae was shown to be polylysogenic . As has been found earlier, the culture has a phage with the original structure of particles whose tail possesses a special substructure called a "collar" . The phage was described in detail as a phage of Bac . thuringiensis 1-97 with a "collar" . Further studies of the culture of Bac . thuringiensis var . galleriae 1-97 has shown that it also contains another phage which differs sharply from the first one in the morphology of particles, the spectrum of lytic action, and other properties . Electron microscopy of the cultural fluid of the Bac . thuringiensis var . galleriae cultures has revealed in them the presence of different amounts of the particles of both morphological types found in the strain 1-97 . These data suggest that the cultures of Bac . thuringiensis var . galleriae are polylysogenic . The methods for the isolation of the new phage are described.

Zh Mikrobiol Epidemiol Immunobiol, 1977 Jul, (7), 111 - 6
{Alteration of the natural humoral immunity in experimental diphtheritic infection induced by different doses of the agent}; Kolisnichenko NI; The authors carried out a complex study of natural humoral immunity factors in rabbits experimentally infected with various doses of diphtheria bacillus (500, 200, 100, 50 and 1 million, 10 000 and 1000 microorganisms) . Natural protection factors--the complement, properdin, and lysozyme proved to undergo similar dynamic changes with each infection dose . The extent and direction of the changes depended on the infection dose . No significant natural immunity changes were caused by a low dose . A dose of 10 000 microbes caused stimulation of properdin and lysozyme activity . Dose of 1, 5, and 50 million suppressed the activity of all the components, the most pronounced with the later dose . Changes occurring with the doses of 200 and 100 million microbes were phasic in character.

Biochem J, 1977 Jul 1, 165(1), 71 - 9
A pulse-radiolysis study of the manganese-containing superoxide dismutase from Bacillus stearothermophilus . A kinetic model for the enzyme action; McAdam ME et al.; The enzymic reaction mechanism of a manganese-containing superoxide dismutase from Bacillus stearothermophilus was studied by using pulse radiolysis . During catalysis (pH 8.9; 25 degrees C), changes occurring in the kinetics of substrate disappearance and in the visible absorption of the enzyme at 480 nm established that the simple two-step mechanism found for copper- and iron-containing superoxide dismutases is not involved . At a low ratio (less than 15) of substrate concentration to enzyme concentration the decay of O2--is close to exponetial, whereas at much higher ratios (greater than 100) the observed decay is predominantly zero-order . The simplest interpretation of the results invokes a rapid one-electron oxidation-reduction cycle ('the fast cycle') and, concurrently, a slower reaction giving a form of the enzyme that is essentially unreactive towards O2-- but which undergoes a first-order decay to yield fully active native enzyme ('the slow cycle') . The fast cycle involves the native enzyme EA and a form of the enzyme EB which can be obtained also by treating the form EA with H2O2 . Computer calculations made with such a simple model predict behaviour in excellent agreement with the observed results.

Am J Trop Med Hyg, 1977 Jul, 26(4), 663 - 78
Intracytoplasmic bacteria in Onchocerca volvulus; Kozek WJ et al.; Ultrastructural studies on Onchocerca volvulus disclosed intracellular organisms within the lateral chords of adult worms and of the larval stages . In the females the organisms were also present in the oogonia, oocytes, developing eggs and microfilariae . The organisms, found within vesicles of host (filarid) membrane and limited to the cytoplasm of infected cells, appeared to have a developmental cycle consisting of three morphologically distinct forms: a small spheroidal form up to 0.3 micronm in size, a bacillary form up to 1.5 micron7 in length and 0.7 micronm in diameter, and a third form, intermediate in size between the former and the latter, characterized by a dense inclusion . The intravesicular location and the developmental cycle consisting of three distinct forms are the two characteristics which suggest that these organisms are more similar to the chlamydiae than to the rickettsiae, in spite of their being transovarially transmitted . The significance of these findings with respect to the host-parasite relationship and pathogenesis of onchocerciasis is presently unknown and will require further study.

Can J Microbiol, 1977 Jul, 23(7), 931 - 2
{Effect of aflatoxin B1 on the enzymatic activities of Bacillus thuringiensis (Berliner)}; Boutibonnes P; At 20 degrees C, aflatoxin B1, at a sublethal dose, decreases the activity of alkaline phosphatase (EC 3.1.3.1), alpha-glucosidase (EC 3.2.1.20), esterase (EC 3.1.1.1), chymotrypsin (EC 3.4.21.1), leucine aminopeptidase (EC 3.4.11.1), and phosphoamidase (EC 3.9.1.1) biosynthesis in Bacillus thuringiensis (Berliner) . In contrast, at 41 degrees C no significant decrease was observed . At this temperature, the mycotoxin is not destroyed or metabolized and bacterial cells are resistant to the toxin.

Mol Biol (Mosk), 1977 Jul-Aug, 11(4), 901 - 8
{Physico-chemical properties of several phages of Bacillus thuringiensis}; Bogush VG et al.; A study was made of biological and physico-chemical properties of phages of Bac . thuringiensis as well as of a number of parameters of nucleic acids isolated from these phages . The phages contain double-stranded DNA . Molecular weights of DNA from three phages--Tg9, Tg10 and Tg13 have been determined by two independent methods: by measuring the contour length of DNA, from the sedimentation constant and for DNA of phage Tg10 also by endonuclease EcRI hydrolysis . These methods gave similar results . On the basis of the temperature of DNA melting the content of GC pairs was found equal to 37.9, 33.4 and 35.1 mole% for DNA's of phage Tg9, Tg10 and Tg13, respectively . On the basis of measuring the intervals of DNA melting a conclusion was made that DNA of the Tg9 and Tg13 phage has a random distribution of base pairs, while DNA of phage Tg10 displays some clustering of base distribution along the molecule . It has been shown that restrictase EcoRI hydrolyses phage Tg10 DNA into 6 fragments of different molecular weights; DNA's of Tg9 and Tg13 phages are not hydrolyzed . A possibility of existance of phage Tg10 DNA in linear and ring forms has been established . The characteristics of phage particles have been determined by electron microscopy.

Ann Sclavo, 1977 Jul-Aug, 19(4), 750 - 7
{Determination of residual antibiotic activity in cerebro-spinal fluid (author's transl)}; Aureli G et al.; A microbiological method for the determination of residual antibiotic activity in CSF is described . This disc-diffusion method, utilizing Bacillus stearotermophilus as test organism, is highly sensitive for many antibiotics, including those crossing the blood-brain barrier.

J Trop Med Hyg, 1977 Jul, 80(7), 144 - 6
A critical review of B.C.G . vaccination programme in Egypt; Madkour M et al.; PIP: Bacille Calmette-Guerin (BCG) vaccination became compulsory in all governorates of Egypt in 1974 . However, a study of 1138 infants attending a private clinic or child welfare center in Cairo found that 624 (55%) had not been vaccinated . There was no significant difference in the vaccination rate between the 2 socioeconomic groups (56% at the private facility and 54% at the public clinic) . 70% of the infants who showed evidence of vaccination and were tuberculin tested had negative results in the determination of postvaccination tuberculin sensitivity . The low rate of vaccination, despite its obligatory nature, appears attributable to concern on the part of both mothers and physicians about the frequency of complications . 90% of pediatricians at the university hospital in Cairo routinely advise their patients against BCG vaccination . The low incidence of tuberculin sensitivity found in this study could not be attributed to social class or racial differences or to an intrinsic defect in the vaccines used . Rather, the poor results are judged to stem from faulty technique of vaccination and/or damage of the vaccine . Administration of intradermal BCG requires a highly trained staff, and use of freeze-dried vaccine is preferable to ensure validity . Independent retesting to evaluate the efficacy of BCG vaccine in randomly selected groups by determining postvaccination conversion rates of tuberculin, vaccination scars, and lymphadenopathy is recommended . Also suggested is the possibility of vaccinating entrants to primary school . The medical service could then have more control over the vaccinations and would be able to monitor more easily the methods used .

Int J Lepr Other Mycobact Dis, 1977 Jul-Sep, 45(3), 221 - 7
Cell-mediated immune response in Mycobacterium leprae infected mice; Navalkar RG et al.; Positive skin reactions to homologous antigens in mice infected with Mycobacterium leprae were observed at the time that the bacillary multiplication was nearing the plateua level . Once developed, the skin reactivity persisted for a considerable length of time . Lymphocyte transformation studies indicated that the lymphocytes from infected animals were unable to respond to specific antigenic stimuli in the early phase of infection . Possible explanation for this failure could be that the lymphocytes although sensitized, were not capable of responding to the stimuli due to a rather low level of sensitization . The elevated lymphocyte stimulation index in the later stages of infection, coupled with skin reactivity indicates the production and proliferation of antigen-reactive lymphocytes which in turn govern the activity of mononuclear phagocytic cells through the production of lymphokines . Occurrence of all the events appears to coincide with the time closer to the plateauing of M . leprae in the foot pad of the mouse . It is suggested that the cell-mediated immune response in the M . leprae infected mouse is basically a T cell rather than a B cell response.

Appl Environ Microbiol, 1977 Jul, 34(1), 7 - 13
Epoxidation of aldrin to exo-dieldrin by soil bacteria; Ferguson JA et al.; Twenty-two strains of soil bacteria, including representatives of the genera Bacillus, Micromonospora, Mycobacterium, Nocardia, Streptomyces, Thermoactinomyces, and Pseudomonas and 10 unidentified gram-negative, motile, rod-shaped bacteria, were shown to degrade aldrin to its epoxide dieldrin . In every case, the exo-stereoisomer of dieldrin was produced exclusively.

Postgrad Med J, 1977 Jul, 53(621), 397 - 9
A case of Bacillus cereus bacteraemia; Barnham M et al.; A case is presented of Bacillus cereus bacteraemia in a patient receiving hepatic perfusion with 5-fluorouracil (5FU) for metastasis from a carcinoma of the breast . The literature concerning systemic B . cereus infections is briefly reviewed.

Cancer Res, 1977 Jul, 37(7 Pt 1), 2251 - 6
Monitoring immunotherapy with Bacillus Calmette-Guérin by antibody titer; Wile AG et al.; Antibody titer to Bacillus Calmette-Guerin (BCG) was determined by complement fixation in 111 normal volunteers and 83 melanoma patients . In 43 of these melanoma patients, sequential determination of antibody titer was made and correlated with clinical course.Low titers of antibody to BCG were found frequently in normal volunteers and in melanoma patients prior to BCG immunotherapy . The typical response to BCG immunotherapy was a rapid rise of antibody titer to high levels . Six of 19 patients with recurrence had disappearance of antibody preceding clinical recurrence by up to 5 months . In 13 of 40 patients receiving BCG, serum antibody was a better indicator of the patient's response than measurement of delayed cutaneous hypersensitivity to purified protein derivative . These results suggest that measurement of the antibody response to BCG may be useful in developing an optimum mode of BCG immunotherapy as well as predicting clinical recurrence in patients with melanoma.

Appl Environ Microbiol, 1977 Jul, 34(1), 102 - 4
Uptake of glucose and maltose by Bacillus popillae; Taylor DC et al.; Results of experiments with glucose and its analog, methyl alpha-D-glucopyranoside, indicated that when glucose was present at low concentrations, it was transported into Bacillus popilliae NRRL B-2309MC cells as glucose 6-phosphate by a phosphoenolpyruvate:sugar phosphotransferase system . An additional mode(s) of entry may be operative at higher glucose concentrations . Maltose appeared to enter the cells by a nonphosphorylative process and was hydrolyzed intracellularly to glucose . No phosphoryl donor was necessary for this hydrolysis.

J Neurosurg, 1977 Jul, 47(1), 35 - 43
Immunobiology of primary intracranial tumors; Mahaley MS Jr et al.; The avian sarcoma virus-induced glioma model in rats was used to study the effectiveness of immunotherapy, namely, Bacillus Calmette-Guerin (BCG) with or without sarcoma cells and/or chemotherapy with 1,3-bis (2-chloroethyl)-1-nitrosourea (BCNU), upon survival times . The most consistent prolongation of survival time was produced by triple therapy: BCG + intraperitoneally administered sarcoma cells + intravenously administered BCNU.

J Bacteriol, 1977 Jul, 131(1), 98 - 104
Bacillus megaterium mutant deficient in membrane-bound adenosine triphosphatase activity; Decker SJ et al.; An adenosine triphosphatase (ATPase) mutant of Bacillus megaterium was isolated and characterized . This mutant (designated A37) was unable to grow on nonfermentable carbon sources and possessed less than 5% of the wild-type ATPase activity . Oxygen uptake by the mutant was comparable to that in the wild type . Sporulation in the wild type occurred in both glucose- and nitrogen-limiting media; however, A37 sporulated only in the nitrogen-limiting medium . The inability of A37 to sporulate in glucose-limiting medium seemed to be due to insufficient adenosine 5'-triphosphate (ATP) levels during the sporulation stages . Fructose, which can generate ATP via substrate-level phosphorylation, is equally efficient in stimulating ATP synthesis in the wild type and A37 . Malate-stimulated ATP synthesis in the wild type was shown to have many characteristics associated with oxidative phosphorylation and was absent in the mutant . These data suggest that the ATPase deficiency results in the loss of oxidative phosphorylation.

Biochem J, 1977 Jul 1, 165(1), 81 - 7
A pulse-radiolysis study of the manganese-containing superoxide dismutase from Bacillus stearothermophilus; McAdam ME et al.; In the preceding paper the mechanism of catalysis of the manganese-containing superoxide dismutase from Bacillus stearothermophilus was shown to involve a 'fast cycle' and a 'slow cycle' {McAdam, Fox, Lavelle & Fielden, 1977 (Biochem . J . 165, 71-79)} . Further properties of the enzyme was considered in the present paper . Pulse-radiolysis studies, under conditions of low substrate concentration to (i.e . when the fast cycle predominates), showed that enzyme activity decreases as pH increases (6.5-10.2) . Activity was unaffected by the addition of H2O2 or NaN3 but slightly decreased by KCN . Both H2O2 and the reducing radical anion CO2-- caused a decrease in A480 of the native enzyme . The rate of the fast catalytic cycle was independent of temperature (5-55 degrees C), and as temperature increases the slow cycle becomes relatively more important . Arrhenius parameters of the rate contants were estimated . The possible identity of the various forms of the enzyme is considered.

Biochim Biophys Acta, 1977 Jun 22, 487(3), 487 - 94
Characteristics of a cytochrome P-450-dependent fatty acid omega-2 hydroxylase from bacillus megaterium; Matson RS et al.; The fatty acid (omega-2) hydroxylase from Bacillus megaterium ATCC 14581 was examined with respect to some general enzymatic properties attributed to an intact complex isolated in a partially purified state . Hydroxylase specific activity was found to increase with increasing protein concentration in a manner consistent with a reversible association of the components in the complex . There was a substantial kinetic lag phase for palmitate hydroxylation which was abolished by a substrate preincubation in the absence of NADPH . The substrate bound and presumably activated the hydroxylase complex without the formation of a substrate-derived intermediated . The oxidation of NADPH and the hydroxylation of palmitate were found to occur in a one to one molar ration, independent of the protein concentration . Finally, a cytochrome P-450 component of the complex was identified on the basis of its CO-binding difference spectrum . It appears, that this cytochrome P-450 component is not identical to P-450 meg of the steroid hydroxylase system of B . megaterium ATCC 13368, since progesterone, an active substrate for the latter, is not hydroxylated by the preparation from B . megaterium ATCC 14581.

Biochem J, 1977 Jun 15, 164(3), 669 - 74
Number and activity of active ribosomes in bacterial polyribosomes; Stenesh J et al.; Polyribosomes were isolated from Baccillus licheniformis, grown at 37 and 46 degrees C, and from Bacillus stearothermophilus, grown at 46 and 55 degrees C . The polyribosomes were incubated with either {3H}puromycin or {14C}phenylalanine . The number of active ribosomes (i.e . those to which growing polypeptide chains are attached) was calculated from the amount of {3H}peptidyl-puromycin formed . The activity of an active ribosome (i.e . the total number of amino acid molecules incorporated/unit time per active ribosome) was calculated from the uptake of {14C}phenylalanine . The number of active ribosomes per migrogram of RNA was as follows: for B . licheniformis, 1.66 X 10(12) and 1.72 X 10(12) at 37 and 46 degrees C respectively; for B . stearothermophilus, 2.59 X 10(12) at 46 and 55 degreesC respectively . The activity per active ribosome was as follows: for B . licheniformis, 0.61 and 0.05 at 37 and 46 degrees C respectively; for B . stearothermophilus, 0.58 and 0.42 at 46 and 55 degrees C respectively.

C R Acad Sci Hebd Seances Acad Sci D, 1977 Jun 13, 284(22), 2301 - 4
{Regulation of isoleucine and valine biosynthesis in Bacillus cereus T: possible role of threonine deaminase}; Raimond J et al.; Threonine desaminase: a possible regulatory element suggested by the analysis of isoleucine, valine pathway regulation in Bacillus cereus T.

Tropenmed Parasitol, 1977 Jun, 28(2), 202 - 4
{Latex-Chagestest-reactions of immunsera against Bartonella bacilliformis, Haemobartonella muris and Eperythrozoon coccoides (author's transl)}; Schottelius J; Sera from 10 patients positive for Carrion's disease were analysed by means of the latex Chagas test . Sera from 40 SPF-rats experimentally infected with Haemobartonella muris and 5 SPF-mice experimentally infected with Eperythrozoon coccoides were similarly tested . Both the human and mice sera were negative whereas positive reaction was observed with rat sera . The nature of this positive reaction has not been clarified.

Can J Microbiol, 1977 Jun, 23(6), 726 - 32
Simultaneous presence of antiviral activity and its degrader in Bacillus extracts; Smart KM et al.; Extracts of uninfected Bacillus species ATCC 27877 cells contained both an antiviral activity and a degrader for this activity . The antiviral activity was stable to 90 degrees C, caused a rapid inhibition of phage adsorption, and resulted in a gradual loss of phage infectivity . When extracts were incubated at 37 degrees C, the degrader inactivated the antiviral activity and the degrader was retained . When the extracts were heated at 90 degrees C, the degrader was inactivated and the antiviral activity was retained . A comparison was made of these two activities in extracts from cells of various incubation ages . The apparent association of maximal antiviral activity with prespore cells seemed due to the high content of degrader in vegetative and stationary cells . Both antiviral activity and its degrader appeared to be characteristics of vegetative cells.

Am J Med, 1977 Jun, 62(6), 949 - 53
Amikacin in the treatment of gram-negative pneumonia; Trenholme GM et al.; A clinical efficacy study of amikacin in the treatment of 15 adults with nonbacteremic, gram-negative bacillary pneumonia is presented . All patients had serious underlying illnesses (11 organic heart disease, five chronic obstructive pulmonary disease, one cancer); 11 had undergone major surgical procedures . All had required respiratory assistance during their hospitalization and all had recently received other antibiotics . Thirteen of 15 patients showed clinical improvement with amikacin therapy; the pathogen was also eradicated in 10 of the 13 . The mean minimum inhibitory concentration of amikacin for the 17 isolated pathogens was 3.13 microng/ml . The mean peak serum concentration of amikacin was 17.7 microng/ml . No evidence of ototoxicity or nephrotoxicity was seen . Seventy-three case reports submitted to the manufacturer by multiple investigators of patients with gram-negative pneumonia, treated with amikacin, are also reviewed . All isolated pathogens were sensitive to both amikacin and gentamicin . Fifty-four (74%) of these patients showed improvement with amikacin therapy.

J Immunol, 1977 Jun, 118(6), 2288 - 90
The effect of BCG on experimental cutaneous leishmaniasis in mice; Weintraub J et al.; The effect of administration of live Bacillus Calmette-Guerin (BCG), a nonspecific immunostimulant, on the course of experimental cutaneous leishmaniasis in mice was investigated . BALB/c mice were injected in the footpad with Leishmania tropica, NIH S-strain; in mice that were not pretreated with BCG this produced a reproducible fatal infection characterized by local inflammation, regional lymphadenopathy, and dissemination of parasites with hepatosplenomegaly . In mice that were pretreated with BCG and similarly infected with L . tropica there was a reduction in the severity of cutaneous disease and a significant (p less than 0.005) decrease in mortality without evidence of visceralization.

J Clin Invest, 1977 Jun, 59(6), 1017 - 26
Effect of chemotherapy and immunotherapy on tumor-specific immunity in melanoma; Mitchell MS et al.; The effects of chemotherapy, with nitrosoureas or dimethyl-triazeno-imidazole-carboxamide (DTIC), or immunotherapy with Bacillus Calmette-Guerin (BCG), on cell-mediated immunity (CMI), and serum blocking factor (BF) to melanoma cells were studied in 23 patients . Studies were performed with autologous or allogenic melanoma target cells obtained from recent biopsy, in 16 mm diameter plastic wells . Assays for lymphocyte-mediated cytotoxicity and BF were performed at weekly intervals over the course of 3-4 mo, with some studies extending beyond 3 yr . The specificity of cytotoxicity was good with these methods . Nine patients given nitrosoureas, predominantly methyl-chloroethyl-cyclohexyl-nitrosourea, showed a transient decline in CMI from 42.2 to 14% 3 wk after administration of a single dose of the agent, with a rapid recovery within 1 week . 10 patients given 5-day courses of DTIC at 3-wk intervals showed no decline in CMI after two courses, and 7 of the 10 had no decline even after three courses . Three of the four patients who achieved a remission lost BF previously present: BF reappeared in both patients studied during a subsequent relapse . BCG intradermally or intralesionally elevated CMI within 2 mo after initiation of therapy, but despite continuation of the injections CMI returned to base line in all but two of the nine patients studied . These results indicate that chemotherapy for melanoma with nitrosoureas or DTIC at these schedules is not profoundly immunosuppressive towards tumor-specific immunity, as measured by our procedures . Putative immunotherapy with BCG at these schedules was likewise only transiently stimulatory.

Acta Pathol Microbiol Scand {B}, 1977 Jun, 85(3), 219 - 26
Inhibition of the synthesis of phospholipase C in Bacillus cereus by a component of the growth medium; Valle KJ et al.; A low molecular weight compound, probably of peptide nature, present in the Beef Heart Extract component of the growth medium, inhibits post-transcriptionally the biosynthesis of phospholipase C by a strain of Bacillus cereus . The compound also prevents the increase of proteolytic activity in the growth medium, thus suggesting that the synthesis of another enzyme, an extracellular protease, is also inhibited, and that the inhibitory compound may therefore have a more general effect on exoenzyme synthesis in this strain of Bacillus cereus.

Zentralbl Bakteriol {Orig A}, 1977 Jun, 238(2), 255 - 62
{Detection of Bacillus cereus-toxins (author's transl)}; Katsaras K et al.; During the logarithmic state of growth, B . cereus produced extracellular toxins, which could be precipitated by ammoniumsulphate . The toxins were not dialysable . 9 strains of B . cereus were tested and showed lethal, hemolytic, and phospholipase-C-reactions . The lethal toxin injected i.v., killed mice within 20 minutes . An intradermal injection of the 9 toxin-precipitates in rabbits produced skin reactions, which could be neutralized by a specific antiserum against the B . cereus strain B-4ac . Relatively high doses (50 mg/ml) of the toxin caused fluid accumulation after injection in ligated loops of rabbit gut (Tab) . The hemolysin of the tested B . cereus strains was found to be thermolabile . The phospholipase-C activity withstood the temperature of 55 degrees C for a period of 10 minutes . The prepared specific antiserum, which inhibited the phsopholipase-C activity totally, had no effect on the hemolysis . Intradermal and enteropathogenic activities point to the presence of an enterotoxin . For detection of the enterotoxin, the skin-test in guinea pigs or rabbits seems to be most appropriate.

Eur J Biochem, 1977 Jun 1, 76(1), 299 - 307
The action of lysozyme on peptidoglycan with N-unsubstituted glucosamine residues . Isolation of glycan fragments and their susceptibility to lysozyme; Amano K et al.; 1 . A peptidoglycan preparation N-acetylated at about 30% of glucosamine residues was obtained by the treatment of the lysozyme-resistant cell wall paptidoglycan of Bacillus cereus with acetic anhydride at pH 7 . Fractionation of dialyzable material resulting from lysozyme digestion of the glycan component of this peptidoglycan preparation yielded five oligosaccharides designated as S1 to S5 besides the disaccharide GlcNAc-MurAc . 2 . Oligosaccharide S3, which accounted for about 30% of the disaccharide units recovered as disaccharides and oligosaccharides, was identified as GlcN-MurAc-GlcNAc-MurAc . Oligosaccharide S1, accounting for about 20% of the disaccharide units recovered, was characterized as GlcN-MurAc-GlcN-MurAc-GlcNAc-MurAc, while oligosaccharide S2, present in a smaller amount, as GlcNAc-MurAc-GlcN-MurAc-glcNAc-MurAc . Oligosaccharides S4, and S5, present in small amounts, were identified as GlcNAc-MurAc-GlcNAc-MurAc and MurAc-GlcNAc-MurAc, respectively . 3 . Oligosaccharides S1, S3 and S5 proved to be completely insusceptible to lysozyme, whereas S2 was digsted by lysozyme to produce GlcNAc-MurAc and S3 . S1 was found to act as a more potent inhibitor than S3 in lysozyme-catalyzed digestion of polysaccharides . 4 . The results obtained show that the lysozyme-catalyzed hydrolysis of peptidoglycan oligosaccharides had an obligatory requirement for the N-acetyl group on the glucosamine residue located in subsite C in the enzyme-substrate complex.

Br J Exp Pathol, 1977 Jun, 58(3), 273 - 80
A severe necrotic enterotoxin produced by certain food, food poisoning and other clinical isolates of Bacillus cereus; Turnbull PC et al.; The ability of certain strains of Bacillus cereus consistently to elaborate a filterable non-dialysable toxin capable of causing severe disruption and necrosis of the intestinal mucosa and submucosa is confirmed . This property is not universal to all B . cereus strains and different degrees of production of this toxin are exhibited by the different strains which produce it . The necrotic effect is produced by whole-cell cultures of the toxin producing strains in broth and in boiled rice . Some characteristics of this necrotic toxin are described and its relationship with the diarrhoeal and other known B . cereus toxins is discussed.

Proc Natl Acad Sci U S A, 1977 Jun, 74(6), 2417 - 21
Membrane ultrastructural changes during calcium phosphate-induced fusion of human erythrocyte ghosts; Zakai N et al.; Nascent calcium phosphate promotes the agglutination and fusion of human erythrocyte ghosts . Membrane phospholipids of erythrocyte ghosts treated with Ca2+ and phosphate ions become exposed to attack by phospholipase C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) (Bacillus cereus) . Freeze-fracture pictures of fused erythrocyte ghosts show the presence of regions deficient in intramemebrane particles in the protoplasmic face which we believe to be regions of fusion . Discontinuous regions of the protoplasmic and exoplasmic faces are observed, which are apparently intermediate stages in the process of fusion . TH-in-section electron micrographs reveal deposits of calcium phosphate in areas of contact and fusion of ghosts . Ca2+ in the presence of N-{tris(hydroxymethyl)methyl}glycine (Tricine) buffer causes the formation of blebs in the membrane but does not cause changes in the intramembrane particle pattern or induce fusion . It is suggested that nascent calcium phosphate acts by forming protein-free regions of phospholipid bilayer which can fuse readily.

Can J Microbiol, 1977 Jun, 23(6), 818 - 23
Phage types of Mycobacterium bovis, substrains of BCG; Mankiewicz E et al.; Nineteen substrains of Mycobacterium bovis, strain bacille Calmette-Guerin (BCG) used in laboratories throughout the world for the preparation of BCG vaccines were phage-typed with a battery of mycobacteriophages . The results revealed differences in their susceptibility to phage lysis that allow subdivision of these strains of BCG into two or more phage types.

J Clin Invest, 1977 Jun, 59(6), 1134 - 42
Circulating immune complexes detected by 125I-Clq deviation test in sera of cancer patients; Teshima H et al.; The presence of circulating immune complexes in freshly drawn sera of patients with various forms of malignancies was detected by the 125I-Clq deviation test of Sobel et al . More than 50% of the 459 cancer sera showed a high inhibition of 125I-Clq uptake by sensitized sheep erythrocytes when compared with sera of 50 healthy laboratory personnel . The levels were compared with levels of total hemolytic complement and immunochemical determinations of Cl1 and C3 . A correlation between high levels of circulating immune complexes and low levels of Clq was suggested . These immune complexes were separated by sucrose density gradient ultracentrifugation at low pH and were found to be heavier than 19S . Fluctuation of levels of immune complexes was evident when serial samples from the same patient were tested . Decrease of levels of immune complexes and a concomitant increase of Clq were detected after Calmette-Gueerin bacillus and autologous tumor cell treatment in some melanoma patients.

Cancer Res, 1977 Jun, 37(6), 1743 - 9
A cytokinetic analysis of bacillus calmette-guérin-induced growth control of a murine leukemia; Olsson L et al.; The cytokinetics of an isogeneic, transplantable, lymphoid leukemia, growing as an ascitic tumor in the C57BL/6 mouse, has been investigated during normal growth and during regression induced by weekly injections i.v . of 1.0 mg Bacillus Calmette-Guerin (BCG) . Survival was significantly prolonged in the BCG-treated group, and 27% of the mice were apparently cured . The tumor growth curves showed, furthermore, that BCG-rreated mice could be divided into two groups according to whether the ascitic tumor cell number was at control level or below that of the controls . By methods such as stathmokinetics, tritiated thymidine autoradiography, and cytophotometry, it was demonstrated that the proliferative activity was higher in BCG mice with a low tumor mass as compared to controls and BCG mice with a tumor mass similar to that of controls . The cytokinetic characteristics of BCG mice with a low ascitic tumor cell number were especially expressed by high mitotic activity, high initial labeling indices, short potential tumor doubling time, and a low number of G0-G1 cells . Furthermore the ascitic tumor cell loss rate was increased in these mice during the whole experimental period . It was deduced from the various parameters and especially from the cytophotometric and autoradiographic results that BCG induces a preferential kill of tumor cells in G0-G1 and the first part of the S phase . In addition, the cytological aspects of the ascitic tumor were found to be related to the cell kinetic pattern as the amount of small and large tumor cells increased and decreased, respectively, with accumulation of tumor cells in G0-G1.

Aust N Z J Surg, 1977 Jun, 47(3), 362 - 5
Complete remission of metastatic malignant melanoma following immunotherapy with Bacillus Calmette-Guerin (BCG): report of a case; Coates AS et al.; A complete response to BCG is described in a case of recurrent melanoma . In a woman aged 38 years, intracutaneous metastatic deposits confined to the limb of origin had occurred after excision of a malignant melanoma from the ankle, and elective groin dissection had shown two lymph nodes infiltrated with melanoma . BCG vaccine was applied to the buttock, initially by scarification and later by a multiple puncture gun . All metastases slowly regressed, and biopsy of a metastatic site at six months showed no tumour cells . The patient remains free of detectable disease 36 months after the commencement of therapy . It is inferred that BCG may facilitate remission of melanoma, perhaps by reason of antigenic cross-reactivity between BCG and surface components of human melanoma cells.

Nucleic Acids Res, 1977 Jun, 4(6), 1829 - 35
The specificity of S1 nuclease toward RNA-DNA hybrids as studied using isotopes of phosphorus-32 and phosphorus-33; Wittelsberger SC et al.; Hybrids were formed from Bacillus cereus DNA and ribosomal RNA . They were treated with various combination of S1 nuclease and ribonuclease, and the molar ratios of the RNA and DNA moieties remaining in the treated hybrids were determined using a 32P-33P dual-label technique . It was found that both S1 nuclease and ribonuclease are required to give hybrid with RNA and DNA in a perfect 1:1 molar ratio . It was noted that the dual-label technique which employs orthophosphate as the sole phosphorus source for both labels gives unambiguous molar ratios and obviates the need to calculate specific activities, make quench corrections, or correct for base content.

Can J Microbiol, 1977 Jun, 23(6), 695 - 700
Use of sepharose-conjugated BCG antibodies for the purification of tuberculin-active components; Laguerre M et al.; Bacillus Calmette-Guerin (BCG) antibodies conjugated with CNBr-activated Sepharose 4B led to the isolation of tuberculin-active components from the protoplasmic materials of BCG . From these results; it is concluded that most of the tuberculin-active components (sensitins) have mycobacterial antigenic properties.

J Bacteriol, 1977 Jun, 130(3), 1010 - 6
Oxygen-dependent inactivation of gramicidin S synthetase in Bacillus brevis; Friebel TE et al.; Incorporation of L-{14C}ornithine into gramicidin S by crude, unfractionated lysozyme extracts of Bacillus brevis ATCC 9999 was shown to represent the activity of the gramicidin synthetase complex . Frozen-thawed cells were the source of active extracts, but when cells were shaken in air at 37 degrees C, they rapidly lost activity in a first-order reaction with a half-life of 13 min . Protease inhibitors and inhibitors of energy metabolism had no effect on the inactivation process in frozen-thawed cells . Stabilization was achieved when the cells were shaken in nitrogen or helium instead of air . The addition of dithiothreitol produced a moderate degree of stabilization . The L-ornithine- and D-phenylalanine-activating activities of the gramicidin S synthetase complex were also lost during aeration of the cells . Crude cell-free extracts also lost activity when they were shaken in oxygen, but, in this case, inactivation was slower (half-life of 80 min) . Nitrogen also stabilized these cell-free extracts.

Antibiotiki, 1977 Jun, 22(6), 486 - 9
{Physicochmemical characteristics of the new antineoplastic antibiotic, nocamycin}; Brazhnikova MG et al.; Nocamycin is produced by Nocardiopsis syringae . It is recovered from the culture fluid by extraction with chloroform . The molecular weight of the crystalline antibiotic is 503, its melting point is 147--149 degrees, {alpha}20 degrees D = --50 degrees (c . 0.21, chloroform), lambdamax235 and 348 nm(E1%sm= = 150 and 420), the summation formula is C26 H33NO9, the biological activity is 100000 Units/mg with respect to Bacillus mycoides . Nocamycin forms salts with alkalies soluble in water . On hydrolysis with an alkali it forms carbonic acid having no ester bond (IP spectrum) and methoxylic group present in the antibiotic molecule . Nocamycin is a new natural substance.

J Biol Chem, 1977 May 25, 252(10), 3252 - 4
Complementation of peptides of barnase, extracellular ribonuclease of Bacillus amyloliquefaciens; Hartley RW; Recovery of ribonuclease activity by complementation of peptides of barnase is reported . Activity is restored to barnase-(1-102), which lacks eight amino acids from its COOH terminus, by combination with peptides-(88-110), -(95-110), or -(99-108), and also with succinylated peptide-(88-110) . The dissociation constants are about 8 X 10(-6) M for the first two combinations and little, if any, greater for the other two . Based on barnase-(1-102) concentration, up to 80% of native activity is obtained with peptide-(88-110) but only 5 to 20% with the others . The octapeptide-(103-110), equivalent to the residues missing from barnase-(1-102), does not complement barnase-(1-102), suggesting that an intact sequence about His-102 and Tyr-103 is required for activity . Barstar, the natural inhibitor of barnase, completely inhibits all activity of the complementing peptides.

J Biol Chem, 1977 May 25, 252(10), 3170 - 5
Inactivation of bacterial D-amino acid transaminase by beta-chloro-D-alanine; Soper TS et al.; Purified D-amino acid transaminase from Bacillus sphaericus catalyzes an alpha,beta elimination from the D isomer of beta-chloroalanine to yield equivalent amounts of pyruvate, chloride, and ammonia; the L isomer of chloroalanine is not a substrate for this transaminase . During the beta elimination there is a synchronous loss in enzyme activity; the Kinact for beta-chloroalanine was estimated to be about 10 micrometers . The alpha-aminoacrylate-Schiff base intermediate formed after beta elimination of chloride ion is probably the key intermediate that partitions between one inactivation event for every 1500 turnovers . In the presence of D-alanine and alpha-ketoglutarate, which are good substrates for the transaminase activity of this enzyme, beta-chloroalanine is a potent, competitive inhibitor (K1 = 10 micrometers) with D-alanine and a weak, uncompetitive inhibitor with alpha-ketoglutarate.

Biochim Biophys Acta, 1977 May 16, 467(1), 44 - 50
Vesicle penicillinase of Bacillus licheniformis 749/C . Apparent identity with the plasma membrane enzyme; Traficante LJ et al.; A substantial fraction of the total membrane penicillinase of Bacillus licheniformis 749/C is attached to the vesicles released during conversion of the cells to protoplasts . This enzyme was purified since there was indirect evidence that it differed from the enzyme that remained with the protoplast . The purified vesicle penicillinase has the same molecular weight and general properties as the plasma membrane (protoplast) enzyme and, similarly, contains a covalently linked phosphatidylserine residue . Treatment of the two enzymes with trypsin produced phosphatidylserine-containing peptides which could not be distinguished by gel or paper electrophoresis . The two membrane penicillinases are very similar, if not identical.

J Mol Evol, 1977 May 13, 9(3), 191 - 201
The rates of evolution in some ribosomal components; Hori H et al.; The rate of nucleotide substitution (k(nuc)) of 5s RNA was estimated to be (1.8 +/- 0.5) x 10(-10) per site per year by comparing the nucleotide sequences of human and Xenopus 5s RNA and using the geological time elapsed since the separation of mammals and amphibians . Similarly, k(nuc) of 5.8s rRNA was calculated to be 0.93 10(-1u) per site per year from the sequences of rat hepatoma cells and Saccbaromyces cerevisiae . For the comparison of these data with the amino acid substitution rate of known proteins, the k(nuc) values of 5s rRNA and 5.8s rRNA were converted to the rate of amino acid substitution (k(aa')) . The k(aa') values in pauling units were 0.4 and 2 0.3, respectively . The average k(aa) of ribosomal proteins was also estimated to be 0.2 0.3 pauling from the N-terminal amino acid sequences of seventeen 30s ribosomal proteins of Bacillus stearothermopbilus and Eschericbia coli . Thus, the evolutionary rates of these ribosomal components studied here are similar to each other; they considerably slower than that of the known cellular proteins . Most, if not all, of the replacements in ribosomal proteins occurred between amino acids of a chemically similar nature.

Arch Microbiol, 1977 May 13, 113(1-2), 23 - 31
{Studies on the thiamine transport system in Bacillus cereus (author's transl)}; Toburen-Bots I et al.; The thiamine transport system in Bacillus cereus exhibits rhythmical changes of resorption- and excretion-phases lasting 1-2 h . These main phases are subdivided in shorter ones with an average duration of 45 s . The velocity of the thiamine uptake is influenced by pH, temperature, age of cells, energy and substrate supply and thiamine concentration of the medium . The Michaelis-Menten-Kinetic can be used to describe the uptake: Km = 1.98 x 10(-8) M; Vmax = 1.19 x 10(-6) mol/g dry weight x min . The rate is enhanced by K+, Ca2+ and Mg2+, and inhibited by Pyrithiamin, EDTA, H+-ions, proton donors and proton acceptors; OH(-)-ions cause a change in the direction of transport . A theoretical explanation can be given by assuming a coupling of the thiamine permeation with proton movements in the membrane.

Vet Pathol, 1977 May, 14(3), 229 - 35
Intestinal Tyzzer's disease and spirochetosis in a guinea pig; McLeod CG et al.; Tyzzer's disease (bacillus pilirformis infection) was diagnosed in young guinea pig killed because of diarrhea and poor condition . There was necrosis and inflammation of the colon, cecum and ileum . Typical B . piliformis were in intestinal epithelial cells . Spirochetes were in large numbers in intestinal crypts and were often adjacent to the intracellular B . piliformis . Extracellular B . piliformis occasionally were surrounded by spirochetes.

Antibiotiki, 1977 May, 22(5), 411 - 3
{Stability of the amidine analogs of penicillin and deacetoxycephalosporin to the penicillinase of Bacillus licheniformis 749/c}; Veinberg GA et al.; Fermentative hydrolysis of 3 derivatives of 6-beta-amidinopenicillanic acid and I derivative of 7-beta-amidinodeacetoxycephalosporanic acid by penicillinase produced by Bacillus licheniformis 749/c was studied . It was found that 6-beta-{(hexahydro-IH-azepin-I-yl) methyleneamino} penicillanic acid, 6-beta-(N1 N-dimethylformamidino-N1) penicillanic acid and 6-beta {(morpholin-I-yl) methylenemino} penicillanic acid were hydrolyzed by the enzyme 50, 70, and 160 times respectively slower than benzylpenicillin . 7-beta-{(Hexahydro-IH-azepin-I-yl)' methylenemino} deacetoxycephalosporanic acid proved to be at least 10 times more stable to the effect of penicillinase than methicillin . In addition unlike the amidine analogues of penicillin the above compound had an inhibitory effect on penicillinase produced by Bacillus licheniformis 749/c.

J Protozool, 1977 May, 24(2), 335 - 40
Reduced pyridine nucleotide oxidases of Eugena gracilis var . bacillaris; Mohanty MK et al.; Cell-free extracts of a streptomycin-bleached strain of Euglena gracilis var . bacillaris have been examined for enzyme systems primarily responsible for the oxidation of reduced pyridine nucelotides . NADH lipoyl dehydrogenase, NADH and NADPH oxidase, NADH and NADPH diaphorase, and NADH and NADPH cytochrome c reductase have been demonstrated . The NADPH-linked enzymes had lower activity rates and were less sensitive to N-ethyl maleimide and p-hydroxymercuribenzoate than their NADH-linked counterparts . NADH cytochrome c reductase was the most sensitive to antimycin A . Michaelis-Menten constants (Km) determined were as follows: NADH diaphorase, 350 muM; NADPH oxidase 150 muM ; NADH lipoyl dehydrogenase, 0.35 muM . Enzyme activities after storage at -5 C indicate that the diaphorases are less labile than the other tested enzymes, and the differential activities of the NADH and NADPH linked enzymes suggest that functionally they may have different roles.

Appl Environ Microbiol, 1977 May, 33(5), 1105 - 11
Study of inhibition of outgrowth in Bacillus cereus T by ethyl picolinate; Pandey NK et al.; The effects of ethyl picolinate on germination, outgrowth, and sporulation of Bacillus cereus T were studied in a synthetic medium containing glucose . Ethyl picolinate specifically inhibited at two stages, outgrowth and sporulation . The initiation of germination and cell division was not affected . The inhibition of outgrowth by ethyl picolinate could be reversed by enrichment of inoculum with aspartic acid, asparagine, lysine, phenylalanine, and tyrosine among the amino acids and by oxalacetate . Nicotinic acid and nicotinamide also possessed this ability . Ethyl picolinate failed to block outgrowth when added to cultures incubated for a short time after inoculation . Enrichment of the medium with lysine plus zinc sulfate stimulated sporulation in the presence of ethyl picolinate to a significance degree.

Proc Natl Acad Sci U S A, 1977 May, 74(5), 1821 - 5
Rapid transmembrane movement of newly synthesized phospholipids during membrane assembly; Rothman JE et al.; The transbilayer distribution of phospholipids in Bacillus megaterium is asymmetrical, with twice as much phosphatidylethanolamine internally as externally (Rothaman, J . E . & Kennedy, E . P . (1977) J . Mol . Biol . 110,603-618) . We now report that the biosynthesis of phosphatidylethanolamine is also asymmetrical . Newly synthesized phosphatidylethanolamine was found first on the cytoplasmic side of the membrane of pulse-labeled cells and later was redistributed until the specific radioactivity of the outer face became equal to that of the inner face of the bilayer . The rate of transmembrane movement is at least 30,000 times faster than the rate of spontaneous diffusion (flip-flop) of phosphatidylethanolamine across artificial phospholipid bilayers, indicating that transmembrane movement must be a facilitated process in living cells, perhaps involving membrane proteins.

Medicine (Baltimore), 1977 May, 56(3), 241 - 54
Gram-negative bacillary pneumonia in the compromised host; Valdivieso M et al.; The clinical and radiological characteristics of 217 consecutive episodes of gram-negative bacillary pneumonia occurring in 189 adult cancer patients between November 1968 and December 1974 were analyzed . The majority of patients had acute leukemia (54%) . Fever larger than or equal to 101 degrees F was the single most common symptom and sign of the presence of infection (90%) . Next in frequency were crepitant rales (65%), cough (41%), dyspnea (19%) and chest pain (18%) . Radiographic evidence of pneumonia was found in 83% of cases and it consisted mainly of alveolar infiltrates involving both lung fields and predominantly the bases . Up to one-third of the patients had normal chestx-ray examinations at the onset of infection, though they subsequently became abnormal in 42% of them . The majority of patients (81%) whose initial chest x-rays did not reveal the presence of pneumonia were neutropenic (less than 1000 circumlating neutrophils/mm3) . Klebsiella sp . and Pseudomonas sp . were the most common infecting organisms . The overall cure rate was 61%; 70% for Klebsiella sp . infections and 64% for Pseudomonas sp . infections . Pulmonary abscesses occurred in 14% of the cases . Cures were related to the antibiotic sensitivity of the infecting organisms and to the number of circulating neutrophils during the period of infection . Best results were obtained with the administration of gentamicin, the newer aminoglycoside antibiotic sisomicin, tobramycin and amikacin, or the combination of gentamicin with carbenicillin or with cephalosporins . Early and vigorous therapy of gram-negative bacillary pneumonia with appropriate antibiotics has improved the prognosis of this infection at our institution.

Infect Immun, 1977 May, 16(2), 542 - 6
Common antigen of Mycobacterium leprae, M . lepraemurium, M . avium, and M . fortuitum in comparative studies using two different types of antisera; Kronvall G et al.; No . 21 mycobacterial antigens of Mycobacterium lepraemurium, M . avium, M . fortuitum, and M . leprae were compared in crossed immunoelectrophoresis using two different antibody sources, a serum pool from lepromatous leprosy patients (LSII) and a rabbit anti-M . smegmatis antiserum . M . lepraemurium, like M . avium, was found to contain the 21 A and 21 C determinants . M . fortuitum contained in addition a new type of determinant, 21 D.M . leprae antigen no . 21 carried the A as well as the B dertminants, the latter found so far only in the leprosy bacillus . The separate taxonomic position of M . leprae, suggested by earlier studies of the no . 21 antigen, is further supported by the present results, which also demonstrate the potential use of submolecular heterogeneity for such investigations.

J Bacteriol, 1977 May, 130(2), 869 - 76
Isolation and characterization of a polynucleotide phosphorylase from Bacillus amyloliquefaciens; Erickson RJ et al.; Bacillus amyloliquefaciens BaM-2 produces large amounts of extracellular enzymes, and the synthesis of these proteins appears to be dependent upon abnormal ribonucleic acid metabolism . A polynucleotide phosphorylase (nucleoside diphosphate:polynucleotide nucleotidyl transferase) was identified, purified, and characterized from this strain . The purification scheme involved cell disruption, phase partitioning, differential (NH4)2SO4 solubilities, agarose gel filtration, and diethylaminoethyl-Sephadex chromatography . The purified enzyme demonstrated the reactions characteristic of polynucleotide phosphorylase: polymerization, phosphorolysis, and inorganic phosphate exchange with the beta-phosphate of a nucleotide diphosphate . The enzyme was apparently primer independent and required a divalent cation . The reactions for the synthesis of the homopolyribonucleotides, (A)n and (G)n, were optimized with respect to pH and divalent cation concentration . The enzyme is sensitive to inhibition by phosphate ion and heparin and is partially inhibited by rifamycin SV and synthetic polynucleotides.

Mikrobiologiia, 1977 May-Jun, 46(3), 580 - 2
{Chemical composition of cells of a chemostatic culture of Bacillus megaterium during exposure to an alkaline pH value}; Sakharova ZV; The culture of Bacillus megaterium was grown under chemostat conditions at a rate of 0.2, 0.4 and 0.7 hr-1, the growth being limited by a low content of citrate, and at alkaline pH values . Critical pH values were obtained for each growth rate: 9.6, 9.2 and 7.6, respectively . The content of protein decreased at alkaline pH, while the synthesis of total lipids, poly-beta-hydroxybutric acid and phospholipids was stimulated . Some changes were found in the ratio between polyphosphates of high molecular weight.

J Biochem (Tokyo), 1977 May, 81(5), 1375 - 81
Two omega-amino acid transaminases from Bacillus cereus; Nakano Y et al.; Bacillus cereus strain K-22 produced two distinct omega-amino acid transaminases, one catalyzing the transamination between beta-alanine and pyruvic acid and the other that between gamma-aminobutyric acid and alpha-ketoglutaric aic . The two enzymes were partially purified and separated from each other by various chromatographies . beta-Alanine:pyruvic acid transaminase and gamma-aminobutyric acid:alpha-ketoglutaric acid transaminase were induced by the addition of beta-alanine and gamma-aminobutyric acid, respectively, to the growth medium . beta-Alanine transaminase showed an optimum pH of 10.0 and optimum temperature of 35 degrees C, and its Km values for beta-alanine and pyruvic acid were both 1.1 mM . gamma-Aminobutyric acid, epsilon-aminocaproic acid, 2-aminoethylphosphonic acid, and propylamine showed about 30-40% of the activity of beta-alanine as amino donors, and oxalacetic acid was as good an amino acceptor as pyruvic acid . The optimum pH and temperature of gamma-aminobutyric acid transaminase were 9.0 and 50 degrees C, respectively, and its Km value for gamma-aminobutyric acid was 2.8 mM, while that for alpha-ketoglutaric acid was 2.3 mM . gamma-Aminobutyric acid and delta-aminovaleric acid were good amino donors but other omega-amino acids were virtually inactive with gamma-aminobutyric acid transaminase; alpha-ketoglutaric acid, and to a lesser extent glyoxylic acid, were active amino acceptors . Sulfhydryl reagents specifically activated gamma-aminobutyric acid transaminase.

J Biochem (Tokyo), 1977 May, 81(5), 1367 - 73
Dihydrodipicolinate reductases from Bacillus cereus and Bacillus megaterium; Kimura K et al.; Dyhydrodipicolinate reductases were purified 100-fold from crude extracts of B . cereus and B . megaterium and their properties were compared with those of the reductase from B . subtilis . The molecular weights of the reductases of B . cereus and B . megaterium were fount to be 155,000 and 150,000, respectively . These reductases were shown to be free of flavin, unlike the B . subtilis enzyme, which contains flavin . Both NADPH and NADH acted as coenzymes for these two reductases . NADPH being three or four times more effective than NADH . The Km values for NADPH and dihydrodipicolinate were 8 micrometer and 62 micrometer, respectively, with B . cereus reductase, and 13 micrometer and 59 micrometer with B . megaterium reductase . The pH optima of the enzymes from B . cereus and B . megaterium were pH 7.4 and 7.2, respectively . The reductases were inhibited by dipicolinate noncompetitively with respect to dihydrodipicolinate and the Ki values were 85 micrometer and 140 micrometer, respectively . Lysine and diaminopimelate were not inhibitory . The properties of the reductases from B . cereus and B . megaterium were similar, but they differed considerably from those of the B . subtilis enzyme . However, all three Bacillus reductases were markedly inhibited by dipicolinate, unlike the enzyme from E . coli.

Biokhimiia, 1977 May, 42(5), 919 - 25
{Purification of phospholipase C from Bacillus cereus by chromatography on aminoalkylpolysaccharide adsorbents}; Gerasimene GB et al.; Purification of phospholipase C from Bac . cereus by chromatography on aminoalkylpolysaccharide adsorbents is described . The dependence of the degree of enzyme purification on the amount of ligant and effect of pH and buffer systems on the adsorption-desorption of phospholipase have been studied . At a pH below 9.0 phospholipase C is not retained by the adsorbents and is purified 4-5-fold and up to 23-fold, when aminoalkyl-Sepharose and hexamethylenediamine Sephadex are used respectively . With an increase in the pH value up to 10.0, the enzyme is bound by the adsorbent and is eluted with a 40-90% yield of activity and 7-10-fold purification . The resulting phospholipase C is highly purified and electrophoretically homogeneous . A mechanism of the enzyme-adsorbent interaction is discussed.

Biochim Biophys Acta, 1977 Apr 26, 487(1), 163 - 74
Membrane lipid metabolism of Bacillus Calmette-Guerin-induced rabbit alveolar macrophages; Wang P et al.; We examined the uptake of radiolabeled lysophospholipids and oleic acid by Bacillus Calmette-Guerin-induced rabbit alveolar macrophages either in the presence or absence of challenge particles . There was no difference in the uptake and metabolism of lysophospholipids by control or challenged cells for incubation periods up to 5 h . When incubated with {3H}oleic acid, challenged cells consistently exhibited a slightly greater uptake of radioactivity . Extraction of the whole cells revealed that the greater amount of radioactivity found in the challenged cells primarily was in triacylglycerol . There was no marked difference in the amount of radioactivity associated with the phospholipids in the whole cell extracts from control and challenged cells . When the macrophages were pre-labeled for 15 min with {3H}oleic acid and then reincubated in fresh medium in the presence or absence of autoclaved Escherichia coli B, more radioactivity was retained by the challenged cells, again in the form of triacylglycerol . Only in isolated plasma membrane fractions did we observe a difference in the amount of radioactivity associated with phospholipids from control and challenged cells . Plasma membranes isolated from Bacillus Calmette-Guerin-induced rabbit alveolar macrophages that had been incubated for 6 h with {3}oleic acid in the presence of E . coli B contained significantly higher level of radioactivity in all lipids than plasma membranes from control cells . Since the greatest and the most consistent difference between control and challenged cells is associated with the triacylglycerol molecule, it is postulated that this molecule may serve as a precursor in the synthesis of alveolar macrophage phospholipids, both by the reacylation pathway and the de novo pathway . It is possible that the high level of radiolabeled phospholipid found in the plasma membrane arose via the de novo pathway following the cleavage of an acyl group as we have found cytidine diphosphocholine phosphotransferase in the plasma membrane fraction (Wang, P., DeChatelet, L.R., and Waite, M . (1977) Biochim . Biophys . Acta 450, 311--321).

Biochemistry, 1977 Apr 5, 16(7), 1355 - 60
Isolation and identification of cytokinins from Euglena gracilis transfer ribonucleic acid; Swaminathan S et al.; Three ribonucleosides responsible for cytokinin activity in Euglena gracilis var Bacillaris tRNA have been isolated and identified as 6-(3-methyl-2-butenylamino)-9-beta-D-ribofuranosylpurine, 6-(4-hydroxy-3-methyl-cis-2-butenylamino)-9-beta-D-ribofuranosylpurine, and 6-(4-hydroxy-3-methyl-2-butenylamino)-2-methylthio-9-beta-D-ribofuranosylpurine . The structures of these compounds were assigned on the basis of their chromatographic properties and ultraviolet and mass spectra which were identical with those of the corresponding synthetic compounds . The elution profiles of cytokinin bioassay activity and of 35S radioactivity suggest the presence of a trace amount of 6-(3-methyl-2-butenylamino)-2-methylthio-9-beta-D-ribofuranosylpurine.

J Biochem (Tokyo), 1977 Apr, 81(4), 813 - 22
Carbohydrate metabolism in Bacillus brevis ATCC 9999; Asatani M et al.; Bacillus brevis ATCC 9999, which produces gramicidin S, was found to be unable to ferment various carbohydrates such as glucose, galactose, mannose, lactose, maltose, sucrose, and mannitol . This defect in carbohydrate utilization was attributed to the lack of the transport system(s) for various carbohydrates . This organism can take up only fructose and glycerol and form acids . The presence in this organism of the phosphoenolpyruvate: sugar phosphotransferase system for fructose, but not glycerol, as well as kinases for fructose and glycerol was demonstrated.

Appl Environ Microbiol, 1977 Apr, 33(4), 878 - 80
Bioassay for homogeneous parasporal crystal of Bacillus thuringiensis using the tobacco hornworm, Manduca sexta; Schesser JH et al.; A method for determining the toxicity of Bacillus thuringiensis subsp . kurstaki parasporal crystal to the tabocco hornworm, Manduca sexta, is described . The use of both mortality and weight loss data have provided a highly sensitive and reproducible bioassay that can be used to compare relative toxicities of crystals from other subspecies as well as toxic components contained therein.

Appl Environ Microbiol, 1977 Apr, 33(4), 865 - 70
Pyrolysis gas-liquid chromatography of the genus Bacillus: effect of growth media on pyrochromatogram reproducibility; Oxborrow GS et al.; Pyrolysis gas-liquid chromatography was performed on dried Bacillus microorganisms to evaluate the effects of growth media . Six cultures of Bacillus and six lot numbers of Trypticase soy agar (BBL) were used to test the hypothesis that a microorganism grown on various lot numbers of the same chromatogram . Also tested was the effect of three different media on chromatogram reproduction using the same six cultures . Results show little or no differences observed between the chromatograms of the individual Bacillus spp . grown on the six lot numbers of Trypticase soy agar . When chromatograms of the three different media were compared, several differences were observed, particularly in the areas most characteristic of individual species . Pryolysis gas-liquid chromatography can be a useful tool for the characterization or identification of the genus Bacillus if the chromatographic and cultural conditions are maintained.

Lab Anim, 1977 Apr, 11(2), 75 - 8
Studies on Tyzzer's disease: isolation and propagation of Bacillus piliformia; Fries AS; Mice from a colony with subclinical Tyzzer's disease were treated with prednisolone in order to activate a Bacillus piliformis infection . From the livers of these mice the organism was isolated and serially passaged in embryonated eggs . It resisted heating at 75 degrees C for 20 minutes and was still infective after storage at -80 degree C for 24 days, but not for 52 days or more . Infectively was maintained for more than a year by storing infectious liver tissue at -190 degrees C . B . piliformis was also isolated from the liver of a mouse with naturally-occurring Tyzzer's disease and maintained for more than 300 passages . Mice inoculated with different egg passages developed fatal Tyzzer's disease and B . piliformis was reisolated from their livers.

J Wildl Dis, 1977 Apr, 13(2), 114 - 6
Tyzzer's disease in wild-trapped muskrats in British Columbia; Chalmers GA et al.; A diagnosis of Tyzzer's disease (Bacillus piliformis infection) was made in a group of recently-trapped muskrats (Ondatra zibethica) . The major lesions consisted of enteritis and focal necrosis of the liver, with organisms resembling B . piliformis in hepatocytes on the periphery of these foci . The outbreak was associated with recent capture, housing, and the use of sulfaquinoxaline-medicated feed.

J Virol, 1977 Apr, 22(1), 194 - 202
Bacteriophage conversion of spore-negative mutants to spore-positive in Bacillus pumilus; Bramucci MG et al.; A pseudolysogenic phage, PMB1, was isolated from soil on the basis of its ability to increase the sporulation frequency of the oligosporogenic Bacillus pumilus strain NRS 576 (sporulation frequency, less than 1%) . Several spore-negative mutants (sporulation frequency, less than 10-8) derived from strain NRS 576, which were converted to spore positive by infection with PMB1, were subsequently identified . PMB1 repeatedly grown on a given spore-negative mutant (e.g., GW2) converted GW2 cells to spore positive . Each plaque-forming unit initiated the conversion of a spore-positive clone in semisolid agar overlays . GW2 cells remained spore positive as long as they maintained PMB1 . Return of PMB1-converted cells to the orginal spore-negative phenotype correlated with loss of PMB1 . In liquid media, PMB1 infection increased the sporulation frequency of mutant GW2 over 106-fold . More than half of the spore-negative mutants we isolated from strain NRS 576 were converted to spore positive by PMB1 infection . PMB1-induced spores of the spore-negative mutant GW2 were somewhat more heat sensitive than uninfected or PMB1-infected spores of the spore positive parent of GW2 . PMB1-induced spores of GW2 do not differ from wild-type spores in morphology by phase-contrast microscopy, dipicolinic acid content, or rate of sedimentation through Renografin gradients.

J Bacteriol, 1977 Apr, 130(1), 375 - 83
Characterization of the entomocidal parasporal crystal of Bacillus thuringiensis; Bulla LA Jr et al.; The parasporal crystalline protoxin of Bacillus thuringiensis contains a single glycoprotein subunit that has a molecular weight of approximately 1.2 X 10(5) . The carbohydrate consists of glucose (3.8%) and mannose (1.8%) . At alkaline pH, the proendotoxin is apparently solubilized and activated by an autolytic mechanism involving an inherent sulfhydryl protease that renders the protoxin insecticidal . Activation generates protons, degraded polypeptides, sulfhydryl group reactivity, proteolytic activity, and insect toxicity . Chemical modification of the sulfhydryl groups inhibits the proteolytic and insecticidal activities, suggesting that cysteine residues may be present in the active site of the protein.

Am J Clin Nutr, 1977 Apr, 30(4), 592 - 8
Weight fluctuations after immunization in a rural preschool child community; Kielmann AA; After inoculations with diphtheria, pertussis, tetanus (DPT), smallpox Bacillus Calmette-Guerin (BCG), polio, and DPT + polio vaccine preparations, weight-for-age fluctuations were monitored in over 470 rural preschool children and compared to those in nonvaccinated control children matched for age, weight-for-age, season and year of immunization, and village affiliation . It was found that children immunized with live agents (BCG, smallpox, polio, DPT + polio) who also were below 6 months of age suffered statistically significant reductions in their weight-for-age compared to matched nonimmunized controls . Children inoculated with polio or smallpox who also were below 80% of the Harvard weight-for-age median experienced a larger decrease in their nutritional levels than those above, with correction for age distribution . It is suggested that in the developing world immunizations with live agents to children below 6 months of age should be given only if the infectious illness in which immunization is provided poses a real threat to health, or if vaccination coverage of children above 6 months of age would subsequently be difficult to achieve.

J Biochem (Tokyo), 1977 Apr, 81(4), 843 - 50
Production of copper coproporphyrin III by Bacillus cereus . II . Regulation of the biosynthesis of coproporphyrin III and its copper complex by oxygen and heavy metal ions; Yamada-Ankei T et al.; The effects of oxygen and heavy metal ions on the production of copper coproporphyrin III were studied in Bacillus cereus strain 2 . The formation of copper coproporphyrin III was found to be maximum when the cells were cultivated in G-medium at a low level of oxygen supply, but it was suppressed at extremely low oxygen supply levels . When the cells were cultured in metal-free G-medium, neither metal-coproporphyrin III nor coproporphyrin III was formed . In the presence of copper in the medium (400-100 micrometers), the formation of coproporphyrin III copper salt was maximum, but the addition of various heavy metal ions other than copper to the copper-free medium resulted in the formation of neither coproporphyrin III nor its metal chelates . Copper ions appear to be specifically required for coproporphyrin III formation.

J Biochem (Tokyo), 1977 Apr, 81(4), 835 - 42
Production of copper coproporphyrin III by Bacillus cereus . I . Purification and identification of copper coproporphyrin III; Yamada-Ankei T et al.; Bacillus cereus strains 2 and T did not form spores and accumulated a large amount of purple pigment inside the cells, when cultured in a yeast extract-ammonium salt medium with excess glucose . The pigment was extracted and crystallized as the ethyl ester . It was identified as copper coproporphyrin III.

J Antibiot (Tokyo), 1977 Apr, 30(4), 283 - 8
Microbial production of vitamin B12 antimetabolites . III Compound 102804 from Bacillus cereus; Kageyama M et al.; A basic compound with empirical formula C12H16N2O5 was isolated from Bacillus cereus 102804 fermentations of a soybean meal-glucose medium . The inhibitory activity of compound 102804 on growth of Gram-positive and Gram-negative bacteria growing in a chemically defined medium was reversed by vitamin B12, by L-methionine, and by D-methionine . It has no inhibitory activity for Escherichia coli (Davis 113-3) when grown in media containing L-methionine . The biosynthesis of 102804 coincides with the sporulation of the B . cereus, and the compound is not produced in the absence of sporulation.

J Bacteriol, 1977 Apr, 130(1), 242 - 8
Role of glutamate in the sporogenesis of Bacillus cereus; Charba JF et al.; Bacillus cereus T, sporulating in a chemically defined medium under optimum conditions, requires substrate quantities of glutamate during the first 4 h of sporogenesis . Seventy percent of the glutamate utilized was catabolized to CO2 during this period, with the remaining glutamate carbon assimilated into various spore constituents, principally protein and nucleic acid . The importance of glutamate as the primary source of reducing potential and energy for early stages of spore formation was investigated . Although the relative efficiency at which tricarboxylic acid cycle intermediates substituted for glutamate was suggestive of oxidation via the tricarboxylic acid cycle, only partial inhibition of glutamate oxidation by fluoroacetate was observed.

J Bacteriol, 1977 Apr, 130(1), 173 - 80
Specificity of siderophore receptors in membrane vesicles of Bacillus megaterium; Aswell JE et al.; Membrane vesicles of Bacillus megaterium strains SK11 and Ard1 bound the ferrischizokinen and ferriferrioxamine B siderhores (iron transport cofactors) . An approximately equimolar uptake of both labels of {3H, 59Fe}ferrischizokinen indicated binding of the intact chelate . Binding reached equilibrium in 2 to 5 min, was temperature independent, and was unaltered by the addition of several energy sources . A 91% dissociation of bound {Fe}ferrischizokinen was achieved in 60 s by the addition of excess ferrischizokinen . Ferriaerobactin, a siderophore which is structurally related to ferrischizokinen, caused no detectable release of bound {59Fe}ferrischizokinen . Of several other ferrigydroxamates tested, only ferriferrichrome A achieved the release (11%) of {Fe}ferrischizokinen . Rapid dissociation (92%) of bound {59Fe}ferriferrioxamine B by the addition of ferriferrioxamine B was observed, and a 67% release of {59Fe}ferriferrioxamine B was caused by ferriA2265, its structural relative . Ferrischizokinen, ferriferrichrome A, and ferrirhodotorulic acid produced a 6, 25, and 29% dissociation, respectively, of {59Fe}ferriferrioxamine B; ferriaerobactin caused no dissociation . {59Fe}ferriaerobactin was bound by the membranes, but its dissociation was not effected by unlabeled ferriaerobactin, suggesting no specific receptors for this chelate . The respective binding affinity constants and maximal binding capacities of membrane vesicles of strain SK11 were 2 x 10(7) M-1 and 280 pmol per mg of protein for ferrischizokinen and 7 x 10(7) M-1 and 37 pmol per mg of protein for ferriferrioxamine B . These values in strain Ard1 were, respectively, 1.4 x 10(7) M-1 and 186 pmol per mg of protein for ferrischizokinen and 11 x 10(7) M-1 and 23 pmol per mg of protein for ferriferrioxamine B . Separate, specific binding sites (receptors) for ferrischizokinen and ferriferrioxamine B exist on the vesicles . The ferrischizokinen receptors have a lower affinity but a higher binding capacity (eightfold) than that shown by the ferriferrioxamine B receptor . These receptors may be components of independent transport systems.

Ann Intern Med, 1977 Apr, 86(4), 456 - 71
Gram-negative rod bacteremia: microbiologic, immunologic, and therapeutic considerations; Young LS et al.; During the last 2 decades, Gram-negative rod bacteremia has become the leading infectious disease problem in American hospitals . With improvements in conventional microbiologic techniques, bacteremic infection can be diagnosed reliably within 3 days using only three sets of cultures . Clinical management still requires aggressive, presumptive use of antimicrobials in patients with the most adverse host factors . In the latter group, the use of combinations of antibiotics that interact synergistically in vitro has improved clinical results . In bacteremia due to anaerobes, particularly Bacteroides species, drainage of infected sites is probably more important than specific drug therapy . Various host defects have been associated with Gram-negative bacteremia; the most common in the nonleukopenic patient is impaired opsonization . The evidence that endotoxins are involved in the pathophysiology of Gram-negative bacillemia is inferential . Nevertheless, both clinical and experimental evidence suggest that active or passive immunization with endotoxin components or antigens similar to Gram-negative polysaccharides may be protective.

Am Rev Respir Dis, 1977 Apr, 115(4), 617 - 23
Mycobacterial antigens relating to experimental pulmonary cavity formation; Maeda H et al.; Lipid-protein mixtures were obtained from 2 strains of mycobacteria, and their cavity-forming activities were examined in rabbit lungs . The mixtures were separated into lipid and protein fractions by gel filtration on Sephadex LH-20 column . Neither lipid nor protein fraction alone had cavity-forming activity; however, restoration of the cavity-forming activity was observed by recombining the fractions . The activity was also reconstructed by combining the protein fraction with cell walls of bacille Calmette-Guerin . The protein fraction from Mycobacterium phlei was further separated into 2 fragments . The larger molecular fragment with a molecular weight of 10,000 daltons consisted of 60% protein and 40% carbohydrate and had cavity-forming activity in combination with the cell walls . The roles of lipid and protein of mycobacteria in cavity formation are discussed.

Pathol Biol (Paris), 1977 Apr, 25(4), 245 - 6
{Report of a case of subcutaneous abscess due to Eikenella corrodens in a splenectomized patient (author's transl)}; Peloux Y et al.; A case of subcutaneous abscess of the chest-wall due to Eikenella corrodens is reported in a patient recently splenectomized . This bacillus was grown in pure culture . Processing specimens and identification methods of this little Gram-negative bacillus are described . Eikenella corrodens is a commensal frequently encountered in mucous membrane surfaces . It is likely that this microorganism may be an "opportunist" pathogen.

Infect Immun, 1977 Apr, 16(1), 213 - 7
Protective role of complement in experimental Escherichia coli endocarditis; Durack DT et al.; Fourteen strains of Escherichia coli were tested for ability to cause infective endocarditis in rabbits prepared by prior placement of an intracardiac catheter . Strains that were resistant to the bactericidal action of serum caused E . coli endocarditis in 91.4% of rabbits, whereas serum-sensitive strains usually failed to cause persisting infection (11.3% infected, P less than 0.001) . Although serum-sensitive E . coli lodged on heart valves within 1 h after intravenous injection, they survived less than 24 h in most normal rabbits . In contrast to normals, all five C6-deficient rabbits injected with a serum-sensitive strain of E . coli developed infective endocarditis (P less than 0.005) . No correlation was found between the presence of K1 antigen and the incidence of experimental E . coli endocarditis . Thus, the ability of strains of E . coli to establish persisting endocardial infection in rabbits appears to be directly associated with resistance to the complement-mediated serum bactericidal system . These findings may explain in part the rarity of gram-negative bacillary endocarditis in patients; they also indicate that in certain special circumstances the serum bactericidal system can play a decisive role in host defense.

Lab Anim, 1977 Apr, 11(2), 69 - 73
Studies on Tyzzer's disease: application of immunofluorescence for detection of Bacillus piliformis and for demonstration and determination of antibodies to it in sera from mice and rabbits; Fries AS; Bacillus piliformis antigens were demonstrated in smear preparations from infected mouse livers by direct immunofluorescence technique . Mouse serum antibodies against B . piliformis were demonstrated by indirect immunofluorescence technique . The test was employed quantitatively both on sera from experimentally infected mice and on sera from clinically healthy mice from colonies infected with B . piliformis, and could be used for the quantitative demonstration of antibodies in sera from a stock of rabbits with Tyzzer's disease . It was found very useful for the detection of subclinical infection.

Acta Hepatogastroenterol (Stuttg), 1977 Apr, 24(2), 102 - 5
Clearance of viable Calmette-Guèrin bacillus by the in vitro isolated and perfused rat liver; Ruggiero G et al.; Liver efficiency in clearing circulating BCG (Clamette-Guerin bacillus) was studied using the isolated rat liver . Bacteria were added to the perfusate at a concentration of 1 X 10(6) cells/ml, and the medium was then sampled at subsequent intervals for 6 hours in 2 perfusions and for 1 hour in 7 perfusions . At the end of all perfusions, liver and bile samples were obtained and used for viable bacterial counts . For each perfusion the bactericidal activity which might have been exerted by serum present in the perfusate was also investigated . About 95% of BCG disappeared from the perfusate after 6 hours of perfusions, and 90% after 1 hour . Recovery of viable bacteria in the liver at 60 minutes averaged 80% of the inoculum . Recovery in bile was negligible . Control experiments indicated that extrahepatic factors in possible reduction of bacterial concentration in the perfusate did not interfere with hepatic removal, per se.

Z Orthop Ihre Grenzgeb, 1977 Apr, 115(2), 249 - 52
{Bone tuberculosis of the bovine type after BCG-vaccination (author's transl)}; Torklus D; It is reported about the BCG-osteomyelitis in early childhood . A long intervall between BCG-vaccination and the onset of clinical symptoms, the good general condition of the patient, nearly unrestricted joint function, often negative bacteriological culture for the tubercle bacillus and negative guinea-pig test are characteristics . Even with extensive lesions of the epiphysis and metaphysis of the long bone and formation of an abscess, the course of the disease seems to be fairly benign . Operation is recommended for getting biopsy material for histological proof of the tuberculous process and for quicker healing.

J Lab Clin Med, 1977 Apr, 89(4), 792 - 803
Tissue factor generation by human mononuclear cells: effects of endotoxin and dissociation of tissue factor generation from mitogenic response; Rickles FR et al.; The effects of the presence of endotoxin in a mononuclear cell culture system have been assessed . Endotoxin was shown to be mitogenic for human peripheral blood lymphocytes and capable of stimulating the generation of tissue factor . Concentrations of endotoxin, found to contaminate many commercial mitogens and antigens, activated mononuclear cells in a time-dependent manner . Generation of tissue factor was detected in cultures harvested from 2 to 72 hours following stimulation with endotoxin . Dose-response curves relating concentrations of endotoxin to mononuclear cell stimulation were determined; as little as 0.001 microng/ml . of E . coli endotoxin was capable of stimulating the generation of tissue factor in the cell cultures . The mitogenic effect of endotoxin was modest, however, and appeared to be unrelated to the ability of endotoxin to active tissue factor . Inhibition of DNA synthesis in the cell cultures by cytosine arabinoside or nonlethal irradiation failed to impair the generation of tissue factor . Endotoxin contamination of various reagents used in cell culture was evaluated with the Limulus assay, which detected as little as 1 X 10(-4) microng/ml . of endotoxin . Endotoxin was detected in preparations of phytohemagglutinin, purified protein derivative of the tubercle bacillus, mumps vaccine, tetanus toxoid, concanavalin A, and pokeweed mitogen . Because of the broad implications of contamination by endotoxin of various reagents, we assessed the specificity of the Limulus assay for the detection of endotoxin in the lectin, concanavalin A, and determined that the reaction was specific for endotoxin . Contamination by endotoxin of mononuclear cell culture systems should be considered as a possible factor in the production of various biological effects attributed to some commonly used mitogens and antigens.

Cancer Res, 1977 Apr, 37(4), 1191 - 6
Effects of methanol extraction residue of Bacillus calmette-Guérin in humans; Perloff M et al.; Forty patients with histologically confirmed neoplastic diseases were treated with the methanol extraction residue of Bacillus Calmette-Guerin (MER) . Thirty-six received concomitant chemotherapy . MER was initially given intradermally twice a month, 1 week apart, at a dose of 200 mug into each of five sites draining different lymph node-bearing areas on the anterior body surface . Thirty-seven patients developed local ulcerations at least 0.5 cm in diameter at MER injection sites . Typical lesion evolution was characterized by erythema and induration followed by vesicle formation and central necrosis . Either granulation tissue or a thick nonulcerated eschar preceded healing, leaving a linear, flat scar . Systemic toxicity consisted of malaise, fever, and myalgias on the day of MER administration . No hematological or biochemical changes directly attributable to MER were observed . Dose titrations in decreasing 10-fold dilutions in a linear array in a single anatomical region were carried out on 35 occasions . All patients but three developed at least a 5-mm induration to the 1-mug dose within 2 weeks of titration . Dose reductions were necessary in 19 instances . The minimal dose that produced a 1-cm inflammatory lesion with central necrosis was 0.01 mug . Serial biopsies were performed . These indicated a time-related series of changes from a nonspecific inflammatory lesion to an acute inflammatory response with microabscesses, followed by noncaseating granulomata and ultimately fibrosis . MER is a quantifiable nonviable immunostimulant that obeys dose-response relationships in its cutaneous lesions.

J Comp Neurol, 1977 Apr 1, 172(3), 381 - 407
Scanning electron microscopy of the subarachnoid space in the dog . V . Macrophages challenged by bacillus Calmette-Guerin; Merchant RE et al.; Mongrel dogs were anesthetized intraperitoneally with pentobarbitol . One cc of cerebrospinal fluid was drawn through a needle inserted into the cisterna magna and mixed with 1 cc (4-9 million viable BCG organisms) of freeze-dried bacillus Calmette-Guerin . One minute later this mixture was injected by the same needle into the cisterna magna . At 1 and 12 days postinjection, experimental animals were perfused with buffered aldehydes . Samples of the leptomeninges were post-fixed in OsO4 and routinely prepared for scanning and transmission electron microscopy . Leptomeningeal samples of untreated, control animals were similarly prepared . Scanning and transmission microscopy confirm that free cells resting on the subarachnoid linings and within the subpial connective tissue space of control animals possess the morphology of macrophages (Malloy and Low, '76) . Viable BCG in the subarachnoid space produces a 3-fold increase in the free cell population of the leptomeninges in 24 hours and a 10-fold increase in 12 days . These cells tend to form associations varying from loose aggregates to tight clusters . Approximately 80% of these free cells express macrophage morphology, with abundant plasma-lemmal microappendages and cytoplasmic vacuoles . Transmission electron microscopy of the free cell population of BCG-stimulated animals reveals at least two other members of the leukocyte series on the leptomeningeal linings.

Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1977 Apr-Jun, 22(2), 87 - 92
{Research on anti-diphtheria immunization in the city of Bucharest}; Spinu I et al.; Biological, sero-epidemiological and bacteriological investigations, carried out in the town of Bucharest for evaluating the present diphtheria vaccination programme showed: -- high anatoxin titers, exceeding the titer considered as protective, in the vaccinates ; -- a concentration of the level of the titers of 1--4 AU/ml, in over 62% of the subjects, which means that the protective titer was exceeded 33 to lucru: -- negative Schick reaction, in the mean 95.06%, in the subjects examined during the 1969--1975 period; -- isolation of the Bacillus diphtheriae in a proportion of 0.08%, none of the toxigenic gravis type . The authors, correlating these data, consider the present vaccination calendar too loaded both as number of inoculations and as antigen concentration per revaccination dose, and recommend reconsideration of the vaccination programme.

Antibiotiki, 1977 Apr, 22(4), 308 - 12
{Processes of spore formation and gramicidin C formation by Bacillus brevis var . G.B.}; Egorov NS et al.; Correlation between gramidicin C biosynthesis and sporulation in the process of Bac . brevis var . G.B . cultivation under various aeration conditions was studied . It was shown that biosynthesis of gramicidin C was characteristic of the young cells and its level was the highest during the culture active growth . The time of the sporulating forms appearance depended on the aeration rate which defined the quantitative composition of the population during the phase of the culture active growth and the stationary phase . Under the optimal aeration conditions the spore formation started during the phase of the culture active growth after some decrease in the maximum level of the cell productivity with respect to the antibiotic . When the aeration rate was increased the spore formation was shifted to later periods of the culture development, i.e . the stationary phase and the phase of the cell autolysis, the gap between the highest levels of gramicidin C buosynthesis and the beginning of sporulation being increased . Under certain aeration conditions the spore formation was not observed, while gramicidin C was synthesized . A conclusion has been made that there is no correlation between gramacidine C biosynthesis and sporualtion in Bacillus brevis var . G.B.

Proc Natl Acad Sci U S A, 1977 Apr, 74(4), 1478 - 82
Biological function of gramicidin: selective inhibition of RNA polymerase; Sarkar N et al.; This paper describes a novel biochemical effect of gramicidin, a class of peptide antibiotics produced by Bacillus brevis during the transition from vegetative growth to sporulation . Gramicidin inhibits RNA synthesis by purified RNA polymerase (nucleosidetriphosphate:RNA nucleotidyl-transferase, EC 2.7.7.6) by interfering with the binding of RNA polymerase to DNA . This effect seems to involve the destabilization of the "open" RNA polymerase-DNA complex, a mode of action consistent with the control of promoter selection . Selectivity in the inhibition of RNA synthesis by gramicidin is observed when transcription is partially blocked by low levels of actinomycin D . Since the inhibition of RNA synthesis by gramicidin is obtained in a highly purified system devoid of membranes, it must be distinct from the ionophoretic activity of the antibiotic . It is possible that this new mode of action reflects the function of gramicidin during bacterial sporulation.

Can J Biochem, 1977 Apr, 55(4), 453 - 7
Reversibility of the ampicillin-and nitrite-induced inactivation of beta-lactamase I; Durkin JP et al.; beta-Lactamase I was isolated from Bacillus cereus 569/H . Treatment with ampicillin in the presence of sodium nitrite at pH 4 or 5 resulted in the inactivation of the enzyme presumably by modification of a carboxyl group in the active site . However, this inactivation was rapidly, reversible at neutral pH and the available evidence points to the participation of a second carboxyl group which is involved in the reactivation process.

Nature, 1977 Mar 24, 266(5600), 328 - 33
Sequence and structure of D-glyceraldehyde 3-phosphate dehydrogenase from Bacillus stearothermophilus; Biesecker G et al.; The glyceraldehyde 3-phosphate dehydogenase holoenzyme of Bacillus stearothermophilus possesses precise 222 symmetry: in this respect it differs from the reported structure of the lobster muscle enzyme . Pairs of active sites are linked through a flexible polypeptide loop which probably mediates the structural changes giving rise to cooperative effects . Three additional salt bridges made by each subunit to others would make a major contribution to thermostability of the tetramer.

J Biol Chem, 1977 Mar 10, 252(5), 1745 - 7
Penicillinase-releasing protease of Bacillus licheniformis 749 Specificity for hydroxyamino acids; Aiyappa PS et al.; The membrane penicillinase of Bacillus licheniformis 749/C differs from the exopenicillinase in that it has an additional 24 amino acid residues and a phosphatidylserine at the NH2 terminus (Yamamoto, S., and Lampen, J.O . (1976) J . Biol . Chem . 251, 4095-4101) . The conversion of the membrane penicillinase to the exo form is probably carried out by a specific penicillinase-releasing protease (PR-protease) whose properties are generally consistent with the properties of penicillinase secretion . The substrate specificity of the PR-protease was determined by identifying the NH2 and COOH termini of the peptides produced by hydrolysis of ribonuclease B and beef insulin . The enzyme hydrolyzed only peptide bonds involving the carboxyl groups of serine or thrombine . Similar bonds in synthetic di- or tripeptides of L-serine were not cleaved . The existence of seryl-lysine and threonyl-glucamic acid bonds in the protease-susceptible (phospholipopeptide) region of the membrane penicillinase and the presence of only lysine or glutamic acid at the NH2 terminus of the exoenzyme released in vivo are consistent with the specificity of PR-protease; hence, we propose that this enzyme has an essential role in the formation of exopenicillinase . The PR-protease is a potential tool for protein sequence determination because of its narrow and novel substrate specificity.

Eur J Biochem, 1977 Mar 1, 73(2), 557 - 65
Specificity profiles of the membrane-bound gamma-D-glutamyl-(L)meso-diaminopimelateendopeptidase and LD-carboxypeptidase from Bacillus sphaericus 9602; Arminjon F et al.; Immunological studies on the carboxypeptidase Y mutant prel-1 of Saccharomyces cerevisiae revealed the origin of mutation in the structural gene of carboxypeptidase Y . The absence of carboxypeptidase Y has no effect on growth, even after drastic changes of growth conditions...

Invest Urol, 1977 Mar, 14(5), 369 - 72
Bacillus Calmette-Guérin and dinitrochlorobenzene immunotherapy of chemically induced bladder tumors; Lamm DL et al.; Immunologic incompetence is correlated with increased susceptibility to malignacy . Immunostimulation with such agents as Bacillus Calmette-Guerin (BCG) and dinitrochlorobenzene (DNCB) have resulted in regression of cutaneous maligancies . An evaluation of the effect of BCG and dinitrochlorobenzene sensitization and bladder irrigation in Fischer rats revealed no decrease in the incidence of bladder tumors induced by 2-formylamino-4-(5-nitro-2-furyl) thiazole (FANFT) . However, direct intralesional injection of BCG into early bladder tumors was effective in preventing the progression of such tumors in seven of ten BCG-sensitized rats (P less than 0.005) . No effect was seen when BCG was injected intralesionally in unsensitized rats or when control solutions were injected.

Cancer Res, 1977 Mar, 37(3), 721 - 5
Characterization of macrophage chemotaxins in tumor cell cultures and comparison with lymphocyte-derived chemotactic factors; Meltzer MS et al.; Culture fluids from five murine sarcomas were chemotactic for syngeneic peritoneal macrophages in vitro . Peritoneal macrophages from mice infected with Mycobacterium bovis, strain Bacillus Calmette-Guerin, were more responsive to the chemotactic factor in tumor cultures than were normal macrophages . Peritoneal granulocytes, however, did not significantly respond to this factor . The level of chemotactic activity in tumor cultures paralleled cell growth for all five tumors; maximal levels occurred during log growth . Culture medium alone or fluids from proliferating spleen cell cultures stimulated with mitogens did not have detectable chemotactic activity . Chromatography of the tumor culture fluids resulted in a single peak of chemotactic activity in the 15,000-molecular weight range on Sephadex G-100 and at about 7.5 mmho/cm specific conductance on diethylaminoethyl cellulose . By both biological and physicochemical characteristics, the chemotactic activity in tumor culture fluids was different from mouse lymphocyte-derived chemotactic factor.

Biochem J, 1977 Mar 1, 161(3), 677 - 85
Enzymic assays for isomers of 2,6-diaminopimelic acid in walls of Bacillus cereus and Bacillus megaterium; Day A et al.; An enzymic assay for individual isomers (meso-, LL- and DD-) of 2,6-diaminopimelate was developed . The enzyme 2,6-diaminopimelate decarboxylase specifically attacked meso-diaminopimelate and was used to measure this isomer manometrically . The meso- and LL-isomers were measured together manometrically in a coupled assay with diaminopimelate decarboxylase and diaminopimelate epimerase (which converts LL-diaminopimelate into meso-diaminopimelate) . The DD-isomer was not attacked by either enzyme and was measured, as residual diaminopimelate after the coupled assay, by a colorimetric method, which was also used to measure total diaminopimelate before enzymic treatments . The coupled enzymes were also used to prepare pure DD-isomer from chemically synthesized diaminopimelate . A mixture of diaminopimelate isomers was present in walls of four strains of Bacillus megaterium {in each about 75% (w/w) meso-, 18% LL- and 7% DD-} and in walls of two strains of Bacillus cereus (about 85% meso-, 8% LL- and 7% DD-) . One strain of B . cereus contained at least 95% meso-diaminopimelate, with only traces of LL- and DD-isomers . Peptidoglycan from Escherichia coli was assayed as containing at least 95% meso-isomer . The proportion of isomers in the wall of a strain of B . megaterium remained constant after growth in a variety of different media.

J Bacteriol, 1977 Mar, 129(3), 1272 - 80
Initiation of Bacillus spore germination by hydrostatic pressure: effect of temperature; Murrell WG et al.; Suspensions of Bacillus cereus T, B . subtilis, and B . pumilus spores in water or potassium phosphate buffer were germinated by hydrostatic pressures of between 325 and 975 atm . Kinetics of germination at temperatures within the range of 25 to 44 degrees C were determined, and thermodynamic parameters were calculated . The optimum temperature for germination was dependent on pressure, species, suspending medium, and storage time after heat activation . Germination rates increased significantly with small increments of pressure, as indicated by high negative deltaV values of -230 +/- 5 cm3/mol for buffered B . subtilis (500 to 700 atm) and B . pumilus (500 atm) spores and -254 +/- 18 cm3/mol for aqueous B . subtilis (400 to 550 atm) spores at 40 degrees C and -612 +/- 41 cm3/mol for B . cereus (500 to 700 atm) spores at 25 degrees C . The ranges of thermodynamic constants calculated at 40 degrees C for buffered B . pumilus and B . subtilis spores at 500 and 600 atm and for aqueous B . subtilis spores at 500 atm were: Ea = 181,000 to 267,000 J/mol; deltaH = 178,000 to 264,000 J/mol; deltaG = 94,000 to 98,300 J/mol; deltaS = 264 to 544 J/mol per degree K . These values are consistent with the concept that the transformation of a dormant to a germinating spore induced by hydrostatic pressure involves either hydration or a reduction in the visocosity of the spore core and a conformational change of an enzyme.

Surv Ophthalmol, 1977 Mar-Apr, 21(5), 429 - 35
Endogenous E . coli endophthalmitis; Shammas HF; A case of Escherichia coli septicemia with associated metastatic en dophthalmitis and endocarditis is presented . The ocular signs and symptoms were the initial manifestations of sepsis . Irreversible damage to the eye occurred in less than 24 hours . The pattern of metastatic bacterial endophthalmitis has changed since the introduction of potent antimicrobial agents, with an increased incidence of Gram-negative bacillemia . E . coli endophthalmitis carries a poor prognosis . Early diagnosis and systemic treatment will prevent the life-threatening complications of sepsis.

Proc Natl Acad Sci U S A, 1977 Mar, 74(3), 999 - 1003
Aminoglycoside-modifying enzyme of an antibiotic-producing bacterium acts as a determinant of antibiotic resistance in Escherichia coli; Courvalin P et al.; Bacillus circulans NRRL B-3312, a nonpathogenic bacterium that produces the aminoglycoside antibiotic butirosin, is known to contain an aminoglycoside phosphotransferase that is similar to the neomycin phosphotransferases of clinically isolated antibiotic-resistant bacteria . Purified DNAs from B . circulans and the plasmid ColE1-ApR were digested with EcoRI endonuclease and the resulting fragments covalently joined with polynucleotide ligase . The recombined DNA was used to transform E . coli and ampicillin-neomycin resistant colonies were selected . Analysis of several clones indicated that neomycin resistance in the E . coli transformants was due to the presence of the B . circulans phosphotransferase gene . This observation is consistent with the notion that anitbiotic-modifying enzymes from antibiotic-producing organisms may be the sources of antibiotic resistance in plasmid-containing bacteria.

Proc Natl Acad Sci U S A, 1977 Mar, 74(3), 1200 - 3
Endotoxin-induced serum factor controlling differentiation of bone-marrow-derived lymphocytes; Hoffmann MK et al.; Serum from bacillus Calmette-Guerin-infected mice injected with endotoxin induces the appearance of surface immunoglobulin, Ia antigen, and complement receptor on the surface of precursor bone-marrow-derived (B) cells . While endotoxin itself causes phenotypic conversion of both thymus-derived (T) cells and B cells in vitro, the endotoxin-induced serum factor was found to be a selective inducer of B cell differentiation . Spleen cells rendered immunodeficient by removal of B cells bearing the complement receptor regained the capacity to cooperate with helper T cells and to produce antibody against red cell antigens in vitro upon upon addition of the serum factor to the culture medium . Thus, a factor that controls selective phenotypic and functional differentiation of B cells has been identified and can now be characterized,

Proc Natl Acad Sci U S A, 1977 Mar, 74(3), 1254 - 8
Alterations of spore coat processing and protein turnover in a Bacillus cereus mutant with a defective postexponential intracellular protease; Cheng YS et al.; A mutant with an alteration in the major intracellular serine protease produced by postexponential Bacillus cereus was isolated by screening mutants defective in spore germination . The purified enzyme from the mutant is more labile to heat and alkaline pH than the protease from the wild type . Protease activity appears at the same time as in the wild type but only reaches 50% of the specific activity and decays more rapidly during sporulation . Coincident with the decay is a decrease in the rate of protein turnover . Generation of amino acids by turnover seems to be important for sporulation because the number of spores produced by the mutant is increased 4- to 10-fold by addition of casamino acids . As anticipated, the mutant produces spores that germinate poorly but, surprisingly, these spores are very deficient in coat protein . Coat antigen is present in cell extracts of mutant and wild type, however, both as large molecules not found on mature spores and as spore coat protein monomers . The large molecules rapidly disappear in a pulse chase experiment in the wild type with some increase in the coat monomers . In mutant extracts, however, this large coat antigen is slowly and improperly processed.

Biochim Biophys Acta, 1977 Feb 7, 459(2), 278 - 89
NMR characterization of three forms of ferredoxin from Desulphovibrio gigas, a sulphate reducer; Moura JJ et al.; A NMR and magnetic susceptibility study of the oxidized and reduced states of three different oligomers (forms) of a {4Fe-4S} ferrodoxin protein from Desulphovibrio gigas, FdI, FdI', and FdII was carried out . FdI and FdI' are different trimers and FdII a tetramer of the same basic subunit . A probable assignment of the contact shifted resonances is indicated . Since the temperature dependences of the contact shifted responances associated with each {4Fe-4S} are not all similar a delocalized model for the spin densities on the 4Fe does not apply . The exchange rate between oxidized and reduced states is slow on the NMR time scale . The three oligomers are not magnetically equivalent . Using the "three state hypothesis" terminology it is shown that FdIox is predominantly in the C2- state and changes upon reduction into the C3- state, while FdIIox is in the C- state and changes into the C2- state . FdI' does not easily fit into this classification . This study shows a similarity of magnetic behaviour between FdI and bacterial ferredoxins (e.g . Bacillus polymyxa) and between FdII and HiPIP from Chromatium sp . The influence of the quaternary structure on the stabilization of the different oxidation states of ferredoxins as well as on their redox potentials is discussed.

Mutat Res, 1977 Feb 1, 46(1), 19 - 26
Cytogenetic hazards from agricultural chemicals . I . A preliminary study on the responses of root meristems to exotoxin from Bacillus thuringiensis a constituent of a microbial insecticide, thuricide; Sharma CB et al.; It reported for the first time that the exotoxin, thuringiensin A, from Bacillus thuringiensis, a component of the insecticide thuricide, inhibits spindle and cytokinesis and induces micronuclei, chromocentric nuclei and minor deviations in spindle activity . The binucleate cells also undergo mitosis yielding biprophases and bimetaphases . Spindle seems to have been inhibited even in bimetaphase . Microtubular systems and chromosomes are implicated as the primary targets . Most effects resemble those of caffeine, colchicine, aminopyrin, chloral hydrate and vinblastine to different extents, and are therefore suggestive of the anti-neoplastic and mutagenic potentialties of the exotoxin . The extensive use of thuricide on crop plants, in view of its mutagenic potential, may be hazardous . The results also suggest that the exotoxin may be used as a pre-treating agent in chromosome analysis and as a candidate-tagging tool for synchronization and cell cycle analysis, besides its probable utility in studies on cancer cells.

J Bacteriol, 1977 Feb, 129(2), 1091 - 101
Bacterial parasite of a plant nematode: morphology and ultrastructure; Sayre RM et al.; The life cycle of a bacterial endoparasite of the plant-parasitic nematode Meloidogyne incognita was examined by scanning and transmission electron microscopy . The infective stage begins with the attachment of an endospore to the surface of the nematode . A germ tube then penetrates the cuticle, and mycelil colonies form in the pseudocoelom . Sporulation is initiated when terminal cells of the mycelium enlarge to form sporangia . A septum within each sporangium divides the forespore from the basal or parasporal portion of the cell . The forespore becomes enclosed by several laminar coats . The parasporal cell remains attached to the forespore and forms the parasporal microfibers . After the newly formed spores are released into the soil, these microfibers apparently enable a mature spore to attach to the nematode . These results indicate that the endoparasite is a procaryotic organism having structural features that are more common to members of Actinomycetales and to the bacterium Pasteuria ramosa than to the sporozoans or to the family Bacillaceae, as previous investigatios have concluded.

J Bacteriol, 1977 Feb, 129(2), 1051 - 8
Role of teichuronic acid in Bacillus licheniformis: defective autolysis due to deficiency of teichuronic acid in a novobiocin-resistant mutant; Robson RL et al.; nov-12, a novobiocin-resistant mutant of Bacillus licheniformis ATCC 9945, grows as long chains of cells, a characteristic of autolytic-deficient (Lyt-) mutants . Isolated walls from nov-12 autolyzed at a rate equal to 5% of that displayed by wild-type walls, thus confirming the Lyt- phenotype . Protein-free nov-12 walls displayed marked resistance to, and also failure to bind, added autolysin solubilized from wild-type walls . Comparison of isolated cell walls revealed a deficiency in teichuronic acid in the mutant . Lesser differences were observed in walls of this strain, including a reduction in galactose, an increase in the proportion of peptidoglycan, and small quantitative differences in peptidoglycan composition though the proportions of protein and teichoic acid were similar in walls of both strains . Autolytic sensitivity was studied in walls in which protein, teichoic acid, and teichuronic acid were removed successively by selective extraction procedures . Autolysis of wild-type walls was unaffected by removal or protein or teichoic acid, but teichuronic acid removal rendered wild-type walls as insensitive to autolysis as mutant walls had been throughout . Therefore, in this mutant, deficiency in teichuronic acid alone leads to the Lyt- phenotype and, hence, activity and binding of autolysin(s) are dependent upon teichuronic acid but not teichoic acid . Also, the potential rate of autolysis of cell walls in this organism was correlated with the proportion of teichuronic acid in the wall . The possible significance of these findings with respect to control of autolysis and cell separation is discussed.

J Bacteriol, 1977 Feb, 129(2), 1045 - 50
Morphological changes associated with novobiocin resistance in Bacillus licheniformis; Robson RL et al.; Spontaneously occurring novobiocin-resistant (Nov) mutants of Bacillus licheniformis ATCC 9945, resistant to low levels of novobiocin (15 mug/ml), were isolated with a frequency of 3 in 106 organisms . Such isolates grew well, but nearly all exhibited consistent plleiotropic alterations in colonial and cell morphologies . One mutant, nov-12, grew as chains of unseparated but clearly distinct daughter cells in the absence of novobiocin in liquid culture . When novobiocin was present, nov-12 grew as very long "filaments" which were, however, septate . Septa formed in the presence of the antibiotic were normal, except that no annular clevage of the septal wall was observed . Septa were also irregularly positioned along the filament . These observations were compared with previous findings on the effects of novobiocin and novobiocin resistance described for other organisms . It was concluded that the primary action of novobiocin might differ in gram-positive and gram-negative organisms . However, when the low-level novobiocin sensitivity, normally associated with gram-positive organisms, was genetically abolished in Nov strains of B . licheniformis they became susceptible to an action of novobiocin more analogous to that found for gram-negative organisms . The morphological alterations associated with the Nov phenotype in this organism, together with observations in other organisms, indicate that novobiocin resistance might be generally useful in the search for mutants of gram-positive organisms with altered cell walls.

J Bacteriol, 1977 Feb, 129(2), 1010 - 9
Biochemical localization of the alkaline phosphatase of Bacillus licheniformis as a function of culture age; Glynn JA et al.; Biochemical localization of the enzyme as a function of age of cell culture showed the alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) activity of Bacillus licheniformis MC14 predominantly in the particulate cell fraction in early- and mid-log cells . However, in late-log and stationary cells, increasing amounts of activity were found in the soluble fraction of lysed cells . Upon protoplast formation of these cells, the activity was released into the soluble fraction . No alkaline phosphatase activity was found in either the cytoplasmic fraction or in the cell medium during any phase of cell growth . The soluble fraction released on protoplast formation that contained alkaline phosphatase activity showed immunological cross-reactivity with antibody to the purified heat--salt-solubilized membrane alkaline phosphatase (F . M . Hulett-Cowling and L . L . Campbell, 1971) . Theparticulate membrane fraction containing a firmly associated alkaline phosphatase also showed similar cross-reactivity . Further, the effectiveness of nonionic detergents, ionic detergents, bile salts, and various concentrations of magnesium and sodium as solubilizing agents for this membrane-bound alkaline phosphatase was investigated . Hexadecyl pyridinium chloride (0.03 M) and magnesium and sodium salts (above 0.2 M) were effective solubilizing agents . The substrate specificities of the various fractions were determined and compared to the substrate specificities of the purified membrane alkaline phosphatase.

J Bacteriol, 1977 Feb, 129(2), 1162 - 4
Density, porosity, and structure of dried cell walls isolated from Bacillus megaterium and Saccharomyces cerevisiae; Scherrer R et al.; Helium displacement and nitrogen adsorption techniques were used to determine the density and porosity, respectively, of freeze-dried cell walls isolated from Bacillus megaterium KM and Saccharomyces cerevisiae . The densities were 1.302 and 1.180 g/cm3, respectively, suggesting noncrystalline solids . The porosities were extremely small, indicating that the cell walls had collapsed and become essentially impervious upon lyophilization.

Eur J Biochem, 1977 Feb, 72(3), 479 - 90
Membrane fractions from the outer layers of spores of Bacillus thuringiensis with toxicity to lepidopterous larvae; Scherrer PS et al.; Two membrane fractions, F1 and F2, have been purified from the outer layers of spores of Bacillus thuringiensis . Both fractions contain 6-7% cysteine and appear to be similar in composition . Amino acids account for about 75% of the dry weight, carbohydrate for about 2% and lipids for about 25% . The fractions are both toxic to Pieris brassicae and the toxicity is inactivated by antiserum to the toxic crystal of Bacillus thuringiensis . The fractions can be distinguished by examination under the electron microscope; both fractions show similar hexagonal patterns but with different spacings . The same fractions from an acrystaliferous mutant (cr) were prepared . These were identical in density and in appearance under the electron microscope; the amino acid analysis of fraction F2 from both strains was identical . However, the spores and fractions F1 and F2 from this strain lacked toxicity . Fraction F2 from the cr strain was used to prepare antiserum specific to fraction F2 . Using this anti-serum and anticrystal serum, crystal and F2 antigens were shown to appear simultaneously in sporulating cultures . Crystal and F2 antigens appeared some time before the maximum rate of uptake of {35s}cysteine . It is concluded that fraction F2 is derived from the exosporium and that fraction F1 probably originates from the spore coat . The exosporium in Bacillus thuringiensis appears to be synthesised during stages II and III of sporulation although uptake of {35S}cysteine occurs much later.

J Hyg (Lond), 1977 Feb, 78(1), 69 - 74
Serotypes of Bacillus cereus from outbreaks of food poisoning and from routine foods; Gilbert RJ et al.; A provisional serotyping scheme was used to type cultures of Bacillus cereus from 84 outbreaks of food poisoning in seven countries; 283 of the 337 (84%) cultures tested were typable . In 35 of the 61 outbreaks associated with a vomiting-type syndrome, foods, clinical specimens or both yielded H-serotype 1 only . Type 1 strains together with other sterotypes were isolated in seven outbreaks . In 14 outbreaks types 3, 4, 5, 8 or a mixture of serotypes were present . Untypable strains were isolated in five outbreaks . Two of the nine diarrhoeal-type outbreaks yielded serotype 1 only . Types 2, 6, 8, 9, 10 and a mixture of type 12 and an untypable strain appeared to be responsible for one outbreak each . Although 16 of the 18 recognized serotypes were present among cultures of B . cereus from various routine foods, only 156 of the 400 (39%) isolates tested were typable.

J Gen Microbiol, 1977 Feb, 98(2), 315 - 27
The enzymic degradation of an alkali-soluble glucan from the cell walls of Saccharomyces cerevisiae; Fleet GH et al.; An alkali-soluble glucan from the cell walls of Saccharomyces cerevisiae NCYC1109 has been hydrolysed with a purified endo-(1 leads to 3)-beta-D-glucanase and an endo-(1 leads to 6)-beta-D-glucanase from Bacillus circulans WL-12 . The products of enzyme action include various oligosaccharide and polysaccharide fractions which have been separated by gel filtration and characterized, giving new information on the fine structure of the glucan . The isolated cell walls have also been subjected to enzymic hydrolysis . The results suggest that part of the cell-wall mannan is held in place by a glucan component.

Cancer, 1977 Feb, 39(2), 570 - 4
Effect of dose, route, and schedule of BCG on antibody titer and survival of murine skin grafts across a weak histocompatibility barrier; Wile AG et al.; Skin was transplanted from male to female C57BL/6JRC mice . Bacillus Calmette-Guerin (BCG) was administered once subcutaneously, intramuscularly, or intraperitoneally either 14 days prior to grafting or on the day of grafting in a dose of 5 X 10(5), 5 X 10(6), or 5 X 10(7) microorganisms . Serum antibody to BCG was assayed using complement fixation . There was a linear inverse relation between skin graft rejection and dose of BCG (p less than .01) such that the lowest dose of BCG was the most effective . The IM and SQ routes were more effective than the IP route . Schedule did not have significant effect on skin graft survival . Time to appearance of antibody correlated directly with dose . The prolonged survival of skin grafts combined with early appearance of antibody to BCG suggested that high doses of BCG could result in antigenic competition.

Cancer Res, 1977 Feb, 37(2), 546 - 50
Direct toxic effects of immunopotentiators on monocytic, myelomonocytic, and histiocytic or macrophage tumor cells in culture; Ralph P et al.; Four murine monocyte, myelomonocyte, and histiocyte or macrophage tumor cell lines adapted to culture were growth inhibited by one or more of the following macrophage-activating substances: Mycobacterium bovis, Bacillus Calmette-Guerin strain, zymosan, lipopolysaccharide, and dextran sulfate, as well as tuberculin purified protein derivative, but not latex beads . Lipopolysaccharide was effective with one line at 4 ng/ml . All four lines actively phagocytosed zymosan and latex beads . In many cases the growth inhibition was apparently immediate but only cytostatic, and cell proliferation resumed upon removal of the drug . Bacillus Calmette-Guerin, live or boiled, was toxic to some of the tumor lines . Synthesis of lysozyme by all the cell lines in the monocyte series and production of granulocyte colony-stimulating factor by the myelomonocytic leukemia were not inhibited during several days of zero growth conditions in the presence of drugs . Since these agents had no direct effect on other hematopoietic tumor types (myeloma, T-lymphoma, mastocytoma) at the same or up to 10(4) higher concentrations, it is proposed that the sensitive tumors retain specific receptors for immunostimulants, either at the cell surface or within the cell in the case of phagocytosable particles . The binding of these agents to physiological receptors leads to stimulation and mitogenesis in normal macrophages and lymphocytes but leads to growth inhibition without affecting differenetiated functions in the corresponding tumor lines.

Ann Microbiol (Paris), 1977 Feb-Mar, 128(2), 151 - 66
{Mechanism of metabolic and ionic germination of "Bacillus licheniformis" spores treated with hydrogen peroxide (author's transl)}; Cerf O; Spores of Bacillus licheniformis 109-2A0 lost their refractility and absorbancy at 640 nm in the presence of metabolizable molecules (L-alanine) . The same occurred with spores treated with 4.4 mol/1 hydrogen peroxide, pH 2.0, at 65 degrees C, even after 5 min of treatment . In addition, these transformations could be promoted after 2 min of treatment by inorganic ions (KI) . This possibility occurs following a kinetics of activation . Thermodynamic parameters showed this activation to be combined with a molecular re-organization . Loss of refractility or absorbancy, induced by L-ala or KI, was inhibited by inhibitors of membrane functions or of L-alanine dehydrogenase, enzyme of which a noticeable activity was demonstrated in treated spores . Only 10% of spore calcium leaked during the treatment . Therefore loss of refractility or absorbancy caused by molecules metabolizable or not seemed to correspond to a physiological germination . The first even of the metabolic, as well as or the ionic germination could well be a modification of the spore membrane proton-motive force.

Proc Natl Acad Sci U S A, 1977 Feb, 74(2), 780 - 4
Biological function of gramicidin: studies on gramicidin-negative mutants; Mukherjee PK et al.; By the use of a rapid radioautographic screening procedure, two mutants of Bacillus brevis ATCC 8185 that have lost the ability to produce gramicidin have been isolated . These mutants produced normal levels of tyrocidine and sporulated at the same frequency as the parent strain . Their spores, however, were more heat-sensitive and had a reduced dipicolinic acid content . Gramicidin-producing revertants occurred at a relatively high frequency among the survivors of prolonged heat treatment and had also regained the ability to produce heat-resistant spores . A normal sport phenotype could also be restored by the addition of gramicidin to cultures of the mutant strain at the end of the exponential growth . On the other hand, the addition of dipicolinic acid could not cure the spore defect . These results provide strong evidence that the inability to produce gramicidin is directly responsible for the observed spore defects . Indeed, they unambiguously demonstrate a function of a peptide antibiotic in bacterial sporulation . The possibility that this function consists of the regulation of transcription during the transition from growth to sporulation is discussed.

Clin Exp Immunol, 1977 Feb, 27(2), 303 - 12
Antigen specific lymphocyte activity in vitro by peripheral blood leucocytes from Mantoux positive and negative human beings . I . Comparison of quantitative and qualitative differences in the PPD-specific lymphoproliferative response of lymphocytes from the two kinds of donors; Jensen B et al.; Lymphocytes from some PPD (purified protein derivative from tubercle bacillus) skin test negative (Mantoux negative=Mx--) human beings reacted against PPD in the lymphoproliferative assay with a time course and dose response very similar to those of lymphocytes from Mantoux positive (Mx+) individuals . Other Mx-- persons were PPD non-responsive in the lymphoproliferative assay . The PPD response of (immunoglobulin=Ig) Ig anti-Ig column passed lymphocytes (T-cells) from Mx--/LP+ (LP+=lymphoproliferative) persons was significantly reduced whereas the in vitro PPD response of T-lymphocytes from Mx+/LP+ was the same or increased . Purified B-lymphocytes from all kinds of tested individuals did not respond in vitro against PPD . Serological investigations indicated that one of the reasons for the negative skin reaction of individuals whose lymphocytes gave a positive lymphoproliferative response against PPD in vitro, is that such individuals had recirculating PPD of high molecular weight (greater than 900,000) and/or PPD anti-PPD antibody complexes in the serum . These substances could block the PPD-specific T-lymphocytes.

J Bacteriol, 1977 Feb, 129(2), 857 - 65
Levels of oxidized and reduced pyridine nucleotides in dormant spores and during growth, sporulation, and spore germination of Bacillus megaterium; Setlow B et al.; Dormant spores of Bacillus megaterium contained no detectable reduced nicotinamide adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) despite significant levels of the oxidized forms of these nucleotides (NAD and NADP) . During the first minutes of spore germination there was rapid accumulation of NADH and NADPH . However, this accumulation followed the fall in optical density that is characteristic of the initiation of spore germination . Accumulation of NADH and NADPH early in germination was not blocked by fluoride or cyanide, and it occurred even when germination was carried out in the absence of an exogenous source of reducing power . In addition to pyridine nucleotide reduction, de novo synthesis also began early in germination as the pyridine nucleotide levels increased to those found in growing cells . Midlog-phase cells grown in several different media had 20 to 35 times as much total pyridine nucleotide as did dormant spores . However, as growth and sporulation proceeded, the NADH plus NAD level fell four- to fivefold whereas the NADPH plus NADP level fell by a lesser amount . From min 10 of spore germination until midway through sporulation the value for the ratio of NADH/NAD is about 0.1 (0.03 to 0.18) while the ratio of NADPH/ANDP is about 1.4 (0.3 to 2.4) . Comparison of these ratios in log-phase versus stationary phase (sporulation) growth in all three growth media tested did not reveal any common pattern of changes.

J Bacteriol, 1977 Feb, 129(2), 1110 - 20
Isolation and properties of a thermostable restriction endonuclease (ENDO R-Bst1503); Catterall JF et al.; A restriction endonuclease was isolated from Bacillus stearothermophilus1503-4R (Bst1503) and purified to homogeneity . The enzyme required Mg2+ ion as a cofactor . Bst1503 exhibited maximal activity between pH 7.5 and 8.0, between 60 and 65 degrees C, and with about 0.2 mM Mg2+ . Bst1503 was not inactivated after exposure at 55 or 65 degrees C for up to 10 h . After 2 h of incubation at 70 degrees C, Bst1503 was inactivated by 65% . Bst1503 was rapidly inactivated at 75 degrees C . A single protein-staining band having a molecular weight of 46,000 was observed when Bst1503 was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The enzyme was found to exist in two active forms, the predominating form with an S value of 8.3 (180,000) and the second form with an S value of 5.4 (96,000) . No conversion between the 8.3S and 5.4S forms was observed after storage . Bst1503 recognized six sites in TP-1C deoxyribonucleic acid (DNA), one site in pSC101 and simian virus 40 DNAs, and three sites in lambdavir DNA . Bst1503 and BamHI were determined to be isoschizomers . The effect of temperatures on the activity and stability of BamHI was determined.

Arch Microbiol, 1977 Jan 11, 111(3), 271 - 82
{Cyclodextrin glucanotransferase from Klebsiella pneumoniae . 1 . Formation, purification and properties of the enzyme from Klebsiella pneumoniae M 5 al (author's transl)}; Bender H; 1 . The strain M 5 al of Klebsiella pneumoniae grows excellently with starches . We were able to show that besides the pullulanase associated with the external membrane of the cells the bacterium produces an inducible, extracellular cyclodextrin glucanotransferase {1,4-alpha-D-glucan-4-alpha-(1,4-alpha-glucano)-transferase (cyclising) (EC 2.4.1.19)} . Potato starch and cyclohexaamylose or cycloheptaamylose were found to be the best "inducing" carbon sources for the synthesis of the enzyme . When the bacteria are grown batchwise, maltose is a poorly "inducing" carbon source; larger quantities of the enzyme are synthesized by continuous cultivation with maltose as growth limiting factor . 2 . For the determination of the cyclodextrin glucanotransferase-activity an assay method wsa worked out . 3 . The enzyme could be separated from the culture filtrate and purified to more than 90% in few steps . At a total yield of 61.2% related to the activity of the culture filtrate employed we received an enzyme solution with the specific activity of 26.6 units/mg protein . Some properties of the enzyme are described . 4 . The products formed from amylopectin by the enzyme were analyzed . Somewhat more than half the amylopectin was found as cyclodextrins . 29.3% of the cyclodextrin fraction were cycloheptaamylose, 47.2% cyclohexaamylose and 10.7% exo-branched cyclohexaamylose . 12.8% of cyclohexaamylose were obtained from a cyclodextrin glucanotransferase-limit dextrin after debranching by pullulanase and exposing the product to the action of the glucanotransferase again . 5 . The importance of the cyclodextrin glucanotransferase for the utilization of starches by this strain of Klebsiella pneumoniae is discussed . After a first characterization the enzyme is compared to the amylase of Bacillus macerans.

Biochim Biophys Acta, 1977 Jan 11, 480(1), 295 - 304
Effect of ADP on ATPASE from a strain of Bacillus stearothermophilus; Hachimori A et al.; Bacillus stearothermophilus ATCC 12016 was unable to grow at temperatures below 40 degrees C . On incubating the bacteria at the temperatures, ATP in cells disappeared, ADP was accumulated and ATPase (EC 3.6.1.3) was inactivated . When the purified ATPase was incubated at the temperatures for 1 h with 0.17 mM ADP in the presence of MgCl2, the enzyme was completely inactivated . The inactivated enzyme was reactivated on dilution or dialysis or on warming at 65 degrees C . During the incubation of the enzyme sample, the absorbance spectrum of the enzyme changed . On further incubating the sample over 1.5 h, the second step of spectral change occurred together with the change of the circular dichrosim and the dissociation into a lower molecular weight species of the protein . When the enzyme was treated with ADP-MgCl2 at 65 degrees C, the inactivation and conformational change of the enzyme was not observed.

Biochim Biophys Acta, 1977 Jan 4, 464(1), 17 - 36
The lipid requirement of the (Ca2+ + Mg2+)-ATPase in the human erythrocyte membrane, as studied by various highly purified phospholipases; Roelofsen B et al.; 1 . When complete hydrolysis of glycerophosphlipids and sphingomyelin in the outer membrane leaflet is brought about by treatment of intact red blood cells with phospholipase A2 and sphingomyelinase C, the (Ca2+ + Mg2+)-ATPase activity is not affected . 2 . Complete hydrolysis of sphingomyelin, by treatment of leaky ghosts with spingomyelinase C, does not lead to an inactivation of the (Ca2+ + Mg2+)-ATPase . 3 . Treatment of ghosts with phospholipase A2 (from either procine pancreas of Naja naja venom), under conditions causing an essentially complete hydrolysis of the total glycerophospholipid fraction of the membrane, results in inactivation of the (Ca2+ + Mg2+)-ATPase by some 80--85% . The residual activity is lost when the produced lyso-compounds (and fatty acids) are removed by subsequent treatment of the ghosts with bovine serum albumin . 4 . The degree of inactivation of the (Ca2+ + Mg2+)-ATPase, caused by treatment of ghosts with phospholipase C, is directly proportional to the percentage by which the glycerophospholipid fraction in the inner membrane layer is degraded . 5 . After essentially complete inactivation of the (Ca2+ + Mg2+)-ATPase by treatment of ghosts with phospholipase C from Bacillus cereus, the enzyme is reactivated by the addition of any of the glycerophospholipids, phosphatidylserine, phosphatidylcholine, phosphatidylethanolamine or lysophosphatidylcholine, but not by addition of sphingomyeline, free fatty acids or the detergent Triton X-100 . 6 . It is concluded that only the glycerophospholipids in the human erythrocyte membrane are involved in the maintenance of the (Ca2+ + Mg2+)-ATPase activity, and in particular that fraction of these phospholipids located in the inner half of the membrane.

Genetika, 1977, 13(3), 496 - 501
{Isolation and preliminary characteristics of the extrachromosomal elements of Bacillus thuringiensis DNA}; Debabov VG et al.; Centrifugation of Bacillus thuringiensis 351 DNA in CsCl-ethidium bromide density gradient has revealed, besides the main band of chromosomal DNA (p = 1.56 g/cm2), some additional bands with higher density, which usually correspond cicle superhelix DNAs, are revealed . 6 discrete bands are observed under electrophoresis of total fraction of minor DNA bands, which suggests, that the preparation contains cicle DNAs of different size . The treatment with DNAse results in the appearance of 3 bands corresponding to opened cicle DNA forms, their molecular weight being 12-10(6)--4-10(6) daltons . Cicle super-helix and opened cicle DNA forms are found in minor fraction by means of electrone microscopy . Statistical analysis has revealed the presence of at least three types of cicle molecules of different size . The variant 351-10, free of extrachromosomal elements, is isolated after ethidium bromide treatment at high temperature . Possible mechanisms of cicle DNAs formation are discussed . Biological function of DNA extrachromosomes fragments in Bacillus thuringiensis is still obscure.

J Biol Stand, 1977, 5(2), 155 - 8
Acceptability of BCG vaccination; Mande R; PIP: Acceptability of Bacille Calmette-Guerin (BCG) vaccine has tended to reflect many factors unrelated to the real effects of the vaccine itself . In particular, the risk of tuberculosis in a given country at a given time has always been a more significant factor than the incidence of complications associated with the vaccine . A possible protective effect of BCG on the occurrence of acute leukemia has also led to the intensification of BCG vaccination among young children entirely on the initiative of physicians . The public is more concerned with anomalous local lesions at the site of intradermal vaccination than by real complications such as fatal dissemination, osteitis, and lupus . As a result, many physicians feel pressured by their patients to replace the intradermal route of administration with scarification and to replace strong strains with weaker strains of vaccine . It is of major importance, then, that public health authorities are correctly informed, with a clear understanding of the balance between the use of an active strain with some side effects but providing good immunity and the use of a weaker strain with fewer side-effects but with a lower level of protection . When tuberculosis is still an important public health problem in a country, relatively innocuous side-effects should be accepted . On the other hand, if tuberculosis is no longer a significant threat, efforts should be made to avoid any annoying consequences of BCG vaccination, no matter how minimal they may be .

Infection, 1977, 5(2), 117 - 8
{A microbiological method for the rapid determination of antibiotic concentrations during therapy (author's transl)}; Wahlig H; The assay described is an agar diffusion test using Bacillus stearothermophilus as test organism . Incubation is at 62 degrees C; results can be obtained rapidly within 2 to 3 hours . The test was developed for the determination of gentamicin concentrations in serum and other body fluids . Because of its high sensitivity the test strain is also suitable for the determination of other antibiotics . The test is designed primarily for the determination of concentrations in individual samples and thus for monitoring during therapy . The ease with which the test can be carried out is thus an important feature of it . On comparison with a standard method (agar diffusion test) using both serum samples from patients and samples with known gentamicin concentrations a highly significant correlation (r = 0.95) was seen between the two methods.

Biochem J, 1977 Jan 1, 161(1), 49 - 62
Folding domains and intramolecular ionic interactions of lysine residues in glyceraldehyde 3-phosphate dehydrogenase; Lambert JM et al.; 1 . Treatment with methyl acetimidate was used to probe the topography of several tetrameric glyceraldehyde 3-phosphate dehydrogenases, in particular the holoenzymes from rabbit muscle and Bacillus stearothermophilus . During the course of the reaction with the rabbit muscle enzyme, the number of amino groups fell rapidly from the starting value of 27 per subunit to a value of approx . five per subunit . This number could be lowered further to values between one and two per subunit by a second treatment with methyl acetimidate . The enzyme remained tetrameric throughout and retained 50% of its initial catalytic activity at the end of the experiment . 2 . Use of methyl {1-14C}acetimidate and small-scale methods of protein chemistry showed that only one amino group per subunit, that of lysine-306, was completely unavailable for reaction with imido ester in the native enzyme . This results is consistent with the structure of the highly homologous glyceraldehyde 3-phosphate dehydrogenase of lobster muscle deduced from X-ray-crystallographic analysis, since lysine-306 can be seen to form an intrachain ion-pair with aspartic acid-241 in the hydrophobic environment of a subunit-subunit interface . 3 . Several other amino groups in the rabbit muscle enzyme that reacted only slowly with the reagent were also identified chemically . These were found to be located entirely in the C-terminal half of the polypeptides chain, which comprises a folding domain associated with catalytic activity and subunit contact in the three-dimensional structure . Slow reaction of these 'surface' amino groups with methyl acetimidate is attributed to intramolecular ionic interactions of the amino groups with neighbouring side-chain carboxyl groups, a conclusion that is compatible with the reported three-dimensional structure and with the dependence of the reaction of ionic stength . 4 . Very similar results were obtained with the enzymes from B . stearothermophilus and from ox muscle and ox liver, supporting the view that the ion-pair involving lysine-306 and aspartic acid-241 will be a common structural feature in glyceraldehyde-3-phosphate dehydrogenases . The B . stearothermophilus enzyme was fully active after modification . 5 . No differences could be detected between the enzymes from ox muscle and ox liver, in accord with other evidence that points to the identify of these enzymes . 6 . The pattern of slowly reacting amino groups in the enzyme from B . stearothermophilus, although similar to that of the mammalian enzymes, indicated one or two additional intramolecular ionic interactions of lysine residues that might contribute to the thermal stability of this enzyme.

Ann Intern Med, 1977 Jan, 86(1), 1 - 5
Unidentified gram-negative rod infection . A new disease of man; Butler T et al.; A Gram-negative bacillus that defies identification was isolated from blood cultures of 17 patients with fever . Fifteen patients were male adults, and 14 patients had underlying diseases, including previous splenectomy in five, which impair host defenses against infection . Illnesses occurred in the summer and autumn in 14 cases and had been recently preceded by dog bites in 10 cases . Clincal syndromes included cellulitis in seven cases, primary bacteremia without localization in four, purulent meningitis in four, and endocarditis in three . Three patients died . The organism grows slowly on blood or chocolate agar in 10% CO, is oxidase- and catalase-positive, and is negative for nitrate reduction, indole production, and urease . It produces acid from glucose, lactose, and maltose . These features distinguish it from all previously described and classified bacteria . Furthermore, the epidemiologic features of the patients suggest that this organism is an opportunistic invader and may have an animal reservoir in nature.

Am J Vet Res, 1977 Jan, 38(1), 81 - 7
Lymphocyte transformation in the dog: response of lymphocytes from normal and immune dogs to phytohemagglutinin, coccidioidin, and purified-protein derivative; Thilsted JP et al.; Antigen- and mitogen-induced in vitro transformation of dog lymphocytes was quantitated by pulse labeling with 3H-thymidine (3H-TdR) . Dosages of antigen and duration of incubation were varied to determine the dose and incubation time that would allow a clear distinction between sensitized and nonsensitized dogs . Each of the 2 antigens tested, coccidioidin and purified protein derivative of tuberculin, induced a higher rate of 3H-TdR incorporation in lymphocyte cultures from dogs immunized to the homologous antigen than in lymphocyte cultures from dogs immunized to either the heterologous antigen or no antigen . Lymphocyte cultures from all dogs showed a high rate of 3H-TdR incorporation in response to phytohemagglutinin stimulation . Sequential lymphocyte transformation tests were done on 4 dogs immunized with either Calmette-Guerin bacillus alone or Calmette-Guerin bacillus in Freund's complete adjuvant . A positive response to purified protein derivative became evident in all 4 dogs 11 days after immunization and peaked at 18 days after immunization.

Can J Surg, 1977 Jan, 20(1), 25 - 30
Adjuvant immunostimulation in malignant melanoma with oral Bacille Calmette-Guérin; MacGregor AB et al.; Results of the administration of oral bacille Calmette-Guerin (BCG) as an adjunct to standard treatment in 62 patients with malignant melanoma indicate that this drug is of value in preventing distant spread of the disease when it is limited to one region and in its early stages . BCG increases survival in patients with visceral metastases . In those in whom these metastases are surgically resectable, it inhibits the development of further metastases . Oral BCG treatment does not affect the course of the disease in patients with massive hepatic or intracranial metastases.

Med Pediatr Oncol, 1977, 3(3), 237 - 42
The phase II evaluation of Bacillus Calmette-Guerin plus megestrol acetate in patients with metastatic renal adenocarcinoma; Laucius JF et al.; The case is reported of a patient with pulmonary metastases from a renal adenocarcinoma who experienced subjective improvement and objective tumor regression on Bacillus Calmette-Guerin (BCG) and megestrol acetate therapy . In a subsequent Phase II trial, no objective responses were noted among 15 patients treated with megestrol acetate (160 mg/day X 56 days) and BCG (five immunizing doses intradermally, every 2 weeks X 5) . It is concluded that this treatment regimen is not clinically useful in patients with metastatic renal adenocarcinoma.

Microbios, 1977, 19(77-78), 231 - 42
Studies on the bacterial spore coat . 6 . Effects of alkali extraction on the spore of Bacillus thiaminolyticus; Minami J et al.; Thin sections of the spore of Bacillus thiaminolyticus Matsukawa and Misawa show a characteristic surface structure with five ridges, and a series of three distinct layers . The outer layer of the spore coat was peeled off by SDS sonic treatment, and then the middle layer was solubilized by alkali extraction of the SDS sonic-treated spore . The spores subjected to these treatments were still refractile, heat resistant, and contained dipicolinic acid, but lost their resistance to mechanical shock.

Ann Nutr Aliment, 1977, 31(4-6), 831 - 40
{Antibacterial properties of aflatoxin B1: cytotoxic effects on Bacilus thuringiensis (Berliner)}; Boutibonnes P et al.; In a sensitive strain of Bacillus thuringiensis (Berliner), aflatoxin B1 inhibited growth with a dose above 5.0 microgram/ml . With subinhibitory levels (0.5 - 5.0 microgram/ml) physiological damage (decreased growth rate) and cellular alteration (filamentous cells) were noted . With these doses the mycotoxin disturbs various metabolisms : DNA synthesis (gradual blockage of the specific bacteriophage multiplication and dispersion of nuclear apparatus in giant cells); protein synthesis (decrease of protease secretion (2.0 microgram/ml), thuricin excretion (3.0 microgram/ml) and inhibition of endotoxin formation (4.0 microgram/ml); specific activities (bacterial motility, flagellar arrangment and sporulation were respectively affected with 1.0, 0.5 and 4.0 microgram/ml) . The mycotoxin did not affect the metabolism of a resistant mutant isolated in the presence of a lethal dose of aflatoxin B1 (mutation rate : 1.10(8) with 20 microgram/ml) . The numerous bacterial responses to aflatoxin B1 indicate that the probable site of its toxic binding may not be restricted to a particular locus on the DNA . On the contrary these observations suggest that there exist many combining affinities of the mycotoxin for intracellular sites or interference with a key-function such as the messenger-RNA synthesis.

Ann Biol Clin (Paris), 1977, 35(4), 325 - 8
{Characterization of the thio-dependent hemolysin of Bacillus alvei}; Kiredjian M; Bacillus alvei hemolytic toxin has been characterized in five main steps from a 18 hours culture supernatant fluid . 50% ammonium sulfate precipitations are followed by Sephadex gel filtrations . After that we proceed to an isoelectric focusing in a narrow pH gradient (from 4 to 6) . A final step consists in a gel filtration allowing to separate sucrose and ampholins from toxin preparation . The homogeneity of the alveolysin preparation is controlled on double immunodiffusion, immunoelectrophoretic analysis and polyacrylamid gel electrophoresis . From this study we can conclude that B . alvei hemolysin has a molecular weight of 45,000 and an isoelectric point of 5.1 . 80 ng/HU of cholesterol represent the 50% inhibitory dose for the hemolytic activity of the alveolysin . Lethal dose calculated on mice is found to be 300 HU.

Ann Nutr Aliment, 1977, 31(4-6), 607 - 15
Toxigenic Aspergilli isolated from different sources in Egypt; Moubasher AH et al.; 45 cultures of Aspergillus, belonging to 25 species isolated from different sources in Egypt, were tested for toxicity to chicken embryos and to Bacillus megaterium growth . Mycotoxins were detected by TLC analysis . 30 isolates out of 45 exhibited toxicity . 3 isolates were recorded as high-toxin producers . (A . egyptiacus, A . carneus and A . terricola) . The 2 former produced unknown toxins, while A . terricola produced aflatoxins (B1, B2, G1 and G2) . Aflatoxins production by A . terricola is reported for the first time.

Microbios, 1977, 18(72), 131 - 40
Studies on the bacterial spore coat 6 effects of alkali extraction on the spore of Bacillus thiaminolyticus; Minami J et al.; Thin sections of the spore of Bacillus thiaminolyticus Matsukawa and Misawa show a characteristic surface structure with five ridges, and a series of three district layers . The outer layer of the spore coat was peeled off by SDS sonic treatment, and than the middle layer was solubilized by alkali extraction of the SDS sonic-treated spore . The spores subjected to these treatments were still refractile, heat resistant, and contained dipicolinic acid, but lost their resistance to mechanical shock.

Tissue Cell, 1977, 9(1), 73 - 85
Studies on the in vivo cellular reactions of insects: an ultrastructural analysis of nodule formation in Galleria mellonella; Ratcliffe NA et al.; Cellular aggregates or nodules which formed in Galleria mellonella larvae in response to injections of killed Bacillus cereus were examined during their first 24 hr of development . The initial aggregation stage was very rapid, occurring within 1 min, and was brought about by the release of an adhesive flocculent material by granular haemocytes in contact with the bacteria . This material, which subsequently became increasingly electron dense owing to melanin deposition, consolidated and by 5 min ramified all the intercellular spaces . The granular cells degenerated, releasing portions of cytoplasm form their peripheries, and eventurally appeared in the matrix as voids containing loosely flocculent material . No bacterial breakdown was seen, however . From 2-4 hr, plasmatocytes began to encapsulate the necrotic mass, and by 24 hr a complete sheath, composed of three discrete regions of cells, was formed . These findings are discussed in relation to the various proposed mechanisms of nodule and capsule formation.

Biochimie, 1977, 59(3), 329 - 36
{Purification of an extracellular thiol-dependent hemolysin from Bacillus alvei}; Alouf JE et al.; Alveolysin a sulfhydryl-dependent cytolytic extracellular protein released by Bacillus alvei has been purified by salting-out by ammonium sulfate, gel filtration, isoelectric focusing on pH gradient and chromatography on DEAE-cellulose . The purified protein after reduction by thiols (active hemolytic form) proved homogeneous by disc polyacrylamide gel electrophoresis and by gel immunodiffusion . The molecular weight was 60,000 daltons, Two molecular forms of pI 5.1 and 7.0 were detected by gel isoelectrofocusing . The toxin was lethal to the mouse . Lytic activity was inhibited by cholesterol and antistreptolysin O anstisera . Immunological cross-reaction was observed between alveolysin and streptolysin O.

Antibiotiki, 1977, 22(2), 136 - 40
{Action of neotelomycin derivatives on bacillus magaterium cells}; Polin AN et al.; The effect of neotelomycin derivatives on the cells of Bac . megaterium was studied . Derivatives with modification of one of the two active centers of the antibiotic molecule, i.e . the free alpha-amine group of the residue of asparaginic acid or hydrophobic triptophanic structure were studied . The derivative with modified indol rings of the residues of beta-methyl-and dehydrotriptophane induced the same though lower damages as the natural antibiotic: increased permeability of the cytoplasmic membranes, protoplast lysis, suppression of the dehydrogenase activity . The activity of this derivative was due to the free amino group and amounted approximately to 3 per cent of the activity of neotelomycin . The derivative with the free amino group of the asparaginic acid residue replaced by the benzoylic group showed a high antibacterial activity but had almost no effect on the membrane permeability and a very low lytic effect . The capacity of this derivative to inhibit the bacterial dehydrogenase activity remained relatively high . Possibly the free amino group of the asparaginic acid residue provided neotelomycin with the capacity for damaging the structure of the bacterial cytoplasmic membranes . No detectable damages in the membrane state after exposure to the benzoylic derivative, as well as its high antibacterial activity are evident of the fact that the mechanism of action of the benzoylic derivative on the cells was in principal different from that of the derivative preserving the free amino group . The triptophane structure was probably not only the center actively affecting the cell but also the factor that provided the antibiotic molecule with conformation most favourable for the action of the free amino group on the membrane structures.

Acta Chem Scand B, 1977, 31(4), 273 - 7
Inactivation of phospholipase C from Bacillus cereus by a carboxyl group modifying reagent; Little C et al.; Phospholipase C from Bacillus cereus was inactivated by incubation with either of the carboxyl reagents, a water-soluble carbodimide plus a nucleophile or Woodward's reagent K . With the former reagent, the incorporation into the enzyme of the first mol of nucleophile caused a 4-5-fold increase in the Km for dihexanoyllecithin with no significant effect on the Vm . The second mol of nucleophile incorporated caused no further change in Km but destroyed most of the catalytic activity . Modification of the enzyme by carbodiimide plus nucleophile did not alter the relative activity of the enzyme towards micelles and monomolecularly dispersed solutions of diheptanoyllecithin . Furthermore, inactivation by this reagent did not significantly decrease the ability of the enzyme to bind to a substrate-based affinity gel . It was concluded that phospholipase C contains a single carboxyl group that is essential for catalytic activity . The enzyme also contains a total of 4-5 reactive/exposed carboxyl groups.

Biochem J, 1977 Jan 1, 161(1), 159 - 65
Phospholipase C from Bacillus cereus . Evidence for essential lysine residues; Aurebekk B et al.; 1 . Phospholipase C was inactivated by exposure to the three amino-group reagents, ethyl acetamidate, 2,4,6-trinitrobenzensulphonic acid and pyridoxal 5'-phosphate plus reduction . 2 . Inactivation by pyridoxal 5'-phosphate showed the characteristics of Schiff's base formation with the enzyme . The pyridoxal 5'-phosphate-treated enzyme after reduction had an absorbance maximum at 325 mm and 6-N-pyridoxyl-lysine was the only fluorescent component after acid hydrolysis . 3 . For complete inactivation, 2 mol of pyridoxal 5'-phosphate or 7 mol of 2,4,6-trinitrophenyl were incorporated/mol of enzyme . 4 . The two apparently essential lysine residues were much more reactive to pyridoxal 5'-phosphate than the other 19 lysine residues in the enzyme . 5 . Binding of phospholipase C to a substrate-based affinity gel caused marked protection against inactivation by pyridoxal 5'-phosphate . For complete inactivation of the gel-bound enzyme, 5 mol of pyridoxal 5'-phosphate were incorporated/mol of enzyme and there was no evidence of two especially reactive lysine residues . 6 . On application of pyridoxal 5'-phosphate-treated enzyme (remaining activity 30% of original) to a column of the affinity gel, some material bound and some did not . The latter contained very little enzyme activity and was heavily incorporated with reagent (9.06 mol/mol of enzyme) . The former had a specific activity of 34% of that of the control and contained 1.29 mol of reagent/mol of enzyme . 7 . Thus phospholipase C appears to contain two lysine residues that are essential for enzyme activity, but probably not for substrate binding.

Folia Microbiol (Praha), 1977, 22(1), 1 - 11
Intracellular proteolytic activity during sporulation of Bacillus megaterium; Chaloupka J et al.; Intracellular proteolytic activity increased during incubation of the sporogenic strain of Bacillus megaterium KM in a sporulation medium together with excretion of an extracellular metalloprotease . The exocellular protease activity in a constant volume of the medium reached a 100-fold value with respect to the intracellular activity . Maximal values of the activity of both the extracellular and intracellular enzyme were reached after 3-5 h of incubation . After 7 h 20-50% cells formed refractile spores . The intracellular proteolytic system hydrolyzed denatured proteins in vitro at a rate up to 150 mug mg-1 h-1 and native proteins at a rate up to 70 mug mg-1 h-1 . Degradation of proteins in vivo proceeded from the beginning of transfer to the sporulation medium at a constant rate of 40 mug mg-1 h-1 and the inactivation of beta-galactosidase at a rate of 70 mug mg-1 h-1 . The intracellular proteolytic activity was inhibited to 65-88% by EDTA, to 23-76% by PMSF . Proteolysis of denatured proteins was inhibited both by EDTA and PMSF more pronouncedly than proteolysis of native proteins; 50-65% of the activity were localized in protoplasts . Another strain of Bacillus megaterium (J) characterized by a high (up to 90%) and synchronous sporulation activity was found to behave in a similar way, but the rate of protein turnover in this strain was almost twice as high . The asporogenic strain of Bacillus megaterium KM synthesized the exocellular protease in the sporulation medium, but its protein turnover was found to decrease substantially after 3-4 h . The intracellular proteolytic system of the sporogenic strain J and the asporogenic strain KM were also inhibited by EDTA and PMSF.

Am Rev Respir Dis, 1977 Jan, 115(1), 151 - 4
Bacillus cereus pneumonia . Survival in a patient with cavitary disease treated with gentamicin; Leff A et al.; Cavitary pneumonitis due to Bacillus cereus occurred in a patient with acute lymphoblastic leukemia . Pneumonia due to this organism is uncommon and recovery has not been previously reported . The present report is the first description of cavitary pneumonia due to B . cereus with a successful outcome . Recovery was attributed to appropriate antimicrobial therapy and induction of leukemic remission with chemotherapy.

Neoplasma, 1977, 24, 225 - 31
Analysis of the results of treament of lung cancer patients with BCG according to the scheme of application of the bacillus; Hadziev S et al.; BCG has successfuly been applied in lymphoid and in myeloid leukemia but no positive results have been demonstrated in solid tumors except malignant melanoma after intratumoral injection . The authors' approach consists in applying BCG as an only antitumor treatment, much smaller doses and in much longer intervals between them than used by other investigators . The analysis of the treatment schemes of BCG, applied to 171 lung cancer patients showed that positive responses have been obtained only in patients without peripheral dissemination of the disease and that in doses from 0.0001 mg to 0.05 mg . The results obtained depended on the frequency of BCG application: the 5 years survival rate, the mean survival period and the rate of the marked X-ray regression have been found best in patients treated once, good in patients treated in intervals longer than 30 days between the applications of the mycobacteria and worse in patients treated in intervals shorter than 20 days . A direct inverse correlation has been discovered between the mean survival period of the treated patients and the number of the applications of BCG in the first 6 months of the treatment.

Am J Anat, 1977 Jan, 148(1), 143 - 8
Identification of challenged subarachnoid free cells; Merchant RE et al.; Three distinct types of free cell contours are recognizable in scanning electron microscopy (SEM) on the leptomeningeal sheaths of dogs twelve days after an intrathecal injection of bacillus of dogs twelve days after an intrathecal injection of bacillus Calmette-Guerin (BCG) . Macrophages posses abundant plasmalemnal blebs which are shown in transmission electron microscopy (TEM) to be composed of large membrane-bound vacuoles . Smooth surfaced lymphoblasts exhibit many basal microvilli that rest upon and often indent the plasmalemma of an underlying pial cell . Neutrophils display many microvilli over their rounded, chrysanthemum-like surfaces . The consistency with which these external features are expressed suggests that each cell type possesses characteristic surface topography, at least under these conditions of challenge.

Cancer Res, 1977 Jan, 37(1), 293 - 8
Changes in distribution of human malignant melanoma membrane antigens in the presence of human antibody by immunofluorescence; Leong SP et al.; Four viable human melanoma cell lines demonstrated full-surface fluorescence (FSF) after incubation at room temperature with antisera from three melanoma patients receiving autologous or homologous immunization with irradiated cultured melanoma cells and Bacillus Calmette-Guerin . By sequential immuno-fluorescent staining, it had been shown that FSF was followed by capping and extrusion of antigen-antibody complexes . One melanoma cell line was tested against its autologous antiserum . In the presence of excess antigen, FSF cells peaked to 25% of the total cell population at 30 min . Maximum capping (20%) was noted at 3 hr . When excess antibody was present, FSF cells were 30% at 2 min and capped cells peaked to 25% at 1.5 hr . Capping ceased at 4 degrees under both conditions . When the serum-treated cells were reincubated after fixation with methanol, 80 to 85% of the cells showed FSF at 2 min to 13.5 hr . Negative controls were preimmune sera and skin fibroblasts . These experimental data suggest the presence of embedded and protruding melanoma membrane antigens in the phospholipid layer . Methanol amy dissolve the lipid layer and expose the embedded antigens . The extrusion of melanoma antigen-human antibody complexes in vitro seems to be a possible mechanism of antigen shedding by melanoma cells in vivo.

J Immunol, 1977 Jan, 118(1), 242 - 7
Lack of interaction of circulating T cells with phytohemagglutinin in bacillary positive untreated lepromatous leprosy patients--identification of subpopulation of lymphocytes by shifts in electrophoretic mobility; Talwar GP et al.; Incubation of human peripheral blood lymphocytes from normal healthy subjects with phytohamagglutinin (PHA), causes the reduction of the surface charge of a subpopulation of T cells by 1363 +/- 242 e.s.u./cm2 . The affected subpopulation was predominantly the high charge-bearing cells identifiable with early (10 min) rosette-forming cells with sheep erythrocytes . Purified lymphocytes obtained from untreated bacillary-positive, lepromatous leprosy patients contained high charge-bearing T lymphocyte subpopulation . However, incubation with PHA did not result in the shift of electrophoretic mobility of these cells, suggesting the absence of interacting sites for the mitogen on the surface of these cells . The absence of mitogen-interacting sites is not an inherent trait of leprosy patients; the surface charge of lymphocytes from Dapsone-treated bacillary-negative subjects was reduced upon incubation with PHA . A close correlation was found between the number of cells whose charge alters on incubation with PHA and the transformation index obtained with this mitogen.

Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Pneumoftiziol, 1977 Jan-Mar, 26(1), 35 - 8
{Immediate results of strictly supervised, intermittent treatment with rifampicin and ethambutol in patients with chronic pulmonary tuberculosis}; Josefovics F et al.; Of the 118 patients suffering from chronic pulmonary tuberculosis, living within the district of the Tirgu Mures Tuberculosis Dispensary, 100 accepted a strictly surveyed treatment and of these 90 continued for at least 6 months . The present paper reports on the immediate radiologic and bacteriological results and the factors influencing them . The following conclusions were drawn: 1) A 2/7 Rifampicin/Etambutol strictly supervised treatment is the most efficient method for neutralizing chronic bacillary sources . 2) The age and origin of the patients, the duration and extent of the pulmonary process, duration of the treatment and associated diseases are the factors that furnish the prognosis of the expected results . 3) Apart from the very good results obtained the method cannot solve all chronic cases and the classical antiepidemic measures must be applied in continuation.

J Bacteriol, 1977 Jan, 129(1), 554 - 5
Presence of cytochrome c in Desulfomonas pigra; Sperry JF et al.; Desulfomonas pigra, a gram-negative, nonmotile anaerobic, sulfate-reducing bacillus isolated from human feces, was found to have cytochrome c and a desulfoviridin-like pigment.

Allerg Immunol (Leipz), 1977, 23(3), 221 - 5
{Immune responses and blood group genetics in patients with asthma bronchiale (author's transl)}; Ksenofontow JP; The combination of MN(Hp 2-2) blood group is more frequent in bronchial asthma patients than in the normal population . The highest changes of blood serum proteinogramm were observed within the first week after anti-typhus-paratyphus-tetanus vaccination in persons with haptoglobulin of 2-2 type, especially in combination with NN and MN blood groups of MN system . The most intense increase of antibody titer to typhoid bacillus was observed in a month after the vaccination in persons with 2-2 type of haptoglobin.

Acta Biol Med Ger, 1977, 36(11-12), 1515 - 22
Relationships between intracellular proteolytic activity and protein turnover in Bacillus megaterium; Chaloupka J et al.; When incubated in a sporulation medium, the sporogenous strains of Bacillus megaterium degrade proteins at a rate of 4-10% X h-1 . The maximal rate of protein turnover is reached after 3-4 hrs at the time of development of forespores and then decreases again . The rate of protein turnover in the asporogenous strain decreases steadily under similar conditions from 3-8% X h-1 at the beginning of incubation to 1% X h-1 after 5-6 hrs in the sporulation medium . The rate of degradation of proteins in vitro in protoplast lysates is similar or higher than the rate of protein turnover . The exocellular, as well as periplasmic proteolytic activity, is suppressed by amino acids more severely than the activity in protoplasts . Mutants devoid of the exocellular proteolytic enzyme contain also less proteolytic activity in the periplasm than in the protoplasts, in contrast to the wild strain . However, their rate of protein turnover, as well as the degradation of abnormal proteins is similar to that in the wild strain . This supports a view that the proteolytic system in protoplasts is involved in intracellular protein catabolism . The periplasmic enzyme can be considered as a kind of the exocellular proteinase.

Vet Med Nauki, 1977, 14(6), 27 - 32
{Gentamycin binding with blood proteins}; Boiadzhieva A et al.; An 'in vitro' microbiologic method was employed to follow up the activity of gentamycin diluted in bovine and swine sera and bound to proteins isolated from them, in the course of continuous storing . It was found that the protein solutions of gentamycin retain their activity for one year under refrigerator conditions (4 degrees C), whereas with the freeze-drying of these solutions storage could be effected at room temperature . It was established through the method of balanced dialysis that gentamycin binds with the proteins of swine sera, and to a largest extent with native serum (30 per cent) and to a lower one--with globulins (15--16.2 per cent) . The protein complexes of gentamycin proved entirely reversible and the process of binding was shown to have no influence on the minimal bacteriostatic concentration of the antibiotic with regard to Bacillus mycoides HB2.

Mikrobiologiia, 1977 Jan-Feb, 46(1), 33 - 7
{Relationship between culture and conditions, the qualitative composition of a culture of Bacillus brevis and its formation of gramicidin C}; Udalova TP et al.; Qualitative composition of the population of Bacillus brevis was found to depend on growth conditions . Under optimum aeration conditions for growth of the culture and biosynthesis of gramicidin C, the population was polymorphous and contained cells and spores of different age . No spores and prospores were found in the culture in conditions of weak and strong aeration . The population consisted only of young, intensively coloured cells with homogeneous or weakly differentiated content in conditions of strong aeration . Cells with the differentiated protoplasm containing large, distinctly seen vacuoles were found in conditions of weak aeration . Gramicidin C was synthesized by young cells with basophilic homogeneous or weakly differentiated protoplasm.

J Biochem (Tokyo), 1977 Jan, 81(1), 269 - 72
Biosynthesis of enzyme-bound formylvaline and formylvalylglycine . A possible initiation complex for gramicidin A biosynthesis; Akashi K et al.; A partially purified fraction (Component I) which catalyzed the ATP-PPi exchange reaction dependent only on valine and glycine among the five constituent amino acids of gramicidin A was obtained from crude extracts of Bacillus brevis ATCC 8185, which produces both gramicidin A and tyrocidines . This fraction was found to bind valine and glycine as thioesters . The addition of a formylTHFA synthesizing system to Component I brought about the formation of Component I-bound formylvaline and formylvalylglycine.

Acta Chem Scand B, 1977, 31(4), 267 - 72
Phospholipase C from Bacillus cereus . Action on some artificial lecithins; Little C; The hydrolysis by phospholipase C from B . cereus of several lecithins of different fatty acyl chain length was examined . The enzyme showed significant activity towards mono-molecularly dispersed short chain lecithins and the reaction obeyed normal Michaelis-Menten kinetics . Rate vs . substrate concentration curves obtained with dihexanoyl-, diheptanoyl- and dioctanoyllecithins showed marked discontinuities in the region of the known critical micelle concentrations for these substrates and distinctly higher rates were obtained just above these levels . Using these three lecithins at levels below their respective critical micelle concentrations, rate increases were noted if the reactions were allowed to proceed to a sufficiently great extent . The presence of deoxycholate in the reaction system had little or no effect on the rate of enzyme-catalysed hydrolysis of lecithins of fatty acyl chain length less than or equal to Cbeta, but for fatty acyl chain lengths greater than C10, significant rate increases occurred . The pH profile for the enzyme activity was also examined.

Mikrobiologiia, 1977 Jan-Feb, 46(1), 15 - 20
{Physiologo-biochemical properties of a chemostatic culture of Bacillus megaterium at different pH values}; Sakharova ZV et al.; Properties of the chemostat culture of Bacillus megaterium were studied during its growth in the state of stress caused by unfavourable acid and alkaline values of pH . The effect of pH is not specific since changes occur in both energy exchange and constructive metabolism . The common action of hydrogen and hydroxyl ions is a decrease in the economic coefficient as greater amounts of the energy substrate are being used, the repression of oxidative enzymes, a decrease in the pool of ATP, and an increase in the respiration quotient . Protein synthesis is inhibited and synthesis of nitrogenless reserve substances is stimulated . Acid and alkaline conditions have also a specific action in the cells of Bac . megaterium . In the alkaline medium, metabolites are liberated into the medium, the cells are enriched with carbohydrates, and acetate is accumulated in the medium . In the acid medium, the cells are enriched with acid insoluble polyphosphates . Idle consumption of the energy substrate takes place.

J Biochem (Tokyo), 1977 Jan, 81(1), 71 - 8
DTNB modification of SH groups of isocitrate dehydrogenase from Bacillus stearothermophilus purified by affinity chromatography; Nagaoka T et al.; A new method of affinity chromatography using blue dextran-Sepharose 4B resin was established to purify NADP+-dependent isocitrate dehydrogenase {EC 1.1.1.42} from Bacillus stearothermophilus in high yield . The purified preparation was found to be homogeneous on disc gel electrophoresis . The SH groups of the enzyme were modified with 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) to determine the number of SH groups per molecule and their contribution to the enzyme activity . One SH group was titrated with DTNB per subunit (the native enzyme consisted of two subunits) and after complete denaturation with 4 M guanidine-HCl the number of titratable SH groups remained unchanged . ORD and CD measurements showed that the alpha-helical conformation of the polypeptide backbone was unaffected by DTNB modification, though the near ultraviolet CD spectrum was evidently altered . The fluorescence derived from tryptophanyl residue(s) was quenched by the modification to 30% of the native level, which may indicate the presence of SH in the vicinity of tryptophanyl residue(s) . A remarkable decrease of the enzyme activity was detected upon modification with DTNB, but there was some discrepancy between the rate of inactivation and that of modification of SH groups . The presence of substrate and Mg2+ gave partial protection against modification of the SH groups by DTNB . Complete protection of the native enzyme activity against heating at 65 degrees was observed in the presence of substrate and Mg2+, but the thermostability of the enzyme was markedly reduced by modification of the SH groups.

Biotechnol Bioeng, 1977 Jan, 19(1), 87 - 99
Production of Schardinger beta-dextrin by soluble and immobilized cyclodextrin glycosyltransferase of an alkalophilic Bacillus sp; Nakamura N et al.; Succinylated cyclodextrin glycosyltransferase (EC 3.2.1.19) of an alkalophilic Bacillus sp . was adsorbed on a vinylpyridine copolymer . The enzyme had about 25% of the activity of soluble enzyme added . No increase of pH or thermal stability of the enzyme was observed by the adsorption, whereas optimum temperature for the enzyme action was shifted from 50 to 55 degrees C . The enzyme converted starch to cyclodextrine without significant loss of activity under the conditions of 4 times reusing of 6 hr conversion by the batch system or 2 weeks continuous reaction by the column system at 55 degrees C and pH 8.0 . About 46% of the potato starch solution {15% (w/v)} was converted to cyclodextrins by the enzyme, and 52% was converted by the simultaneous action of the enzyme and alkaline pullulanase of alkalophilic Bacillus sp . (No . 202-1) . These values were almost the same as those obtained by the soluble enzyme or enzymes system.

J Bacteriol, 1977 Jan, 129(1), 191 - 7
Penicillinase-releasing protease of Bacillus licheniformis: purification and general properties; Aiyappa PS et al.; The membrane penicillinase of Bacillus licheniformis 749/C is a phospholipoprotein which differs from the exoenzyme in that it has an additional sequence of 24 amino acid residues and a phosphatidylserine at the NH2 terminus . In exponential-phase cultures, the conversion of membrane penicillinase to exoenzyme occurs at neutral and alkaline pH . An enzyme that will cleave the membrane penicillinase to yield the exoenzyme is present (in small amounts) in exponential-phase cells and is released during their conversion to protoplasts . The enzyme is found in the filtrate of a stationary-phase culture of the uninduced penicillinase-inducible strain 749 and has been purified to apparent homogeneity from this source . The protease has an approximate molecular weight of 21,500 and requires Ca2+ ions for stabilization . It has a pH optimum of 7.0 to 9.5 for hydrolysis of casein and for the cleavage of membrane penicillinase . Both activities are inhibited by diisopropylfluorophosphate; hence, the enzyme is a serine protease . This enzyme may be entirely responsible for the formation of exopenicillinase by this organism, since the other neutral and alkaline proteases of strain 749 have little, if any, activity in releasing exopenicillinase . The enzyme has been termed penicillinase-releasing protease.

J Bacteriol, 1977 Jan, 129(1), 184 - 90
Vesicle penicillinase of Bacillus licheniformis: existence of periplasmic-releasing factor(s); Traficante LJ et al.; In earlier studies of the membrane-bound penicillinase of Bacillus licheniformis 749/C, the enzyme present in the vesicles that were released during protoplast formation and the enzyme retained in the plasma membrane of protoplasts appeared to differ (i) in their behavior on gel permeation chromatography in the presence or absence of deoxycholate and (ii) in their tendency to convert to the hydrophilic exoenzyme (Sargent and Lampen, 1970) . We have now shown that these vesicle preparations contain a soluble, heat-sensitive enzyme(s) that is released along with the vesicles during protoplast formation . The enzyme will convert the vesicle penicillinase to a form that resembles exopenicillinase, and this conversion can be inhibited by deoxycholate under certain circumstances . Sedimentation of such vesicle preparations at 100,000 X g produces vesicles which contain penicillinase that behaves as the plasma membrane enzyme obtained from protoplasts . Exopenicillinases released by growing cells at pH 6.5 and by washed cells or protoplasts at pH 9.0 have the same NH2-terminal residues (lysine and some glutamic acid); in addition, the various release systems show a parallel sensitivity to inhibition by deoxycholate, quinacrine, chloroquine, and o-phenanthroline . The formation of exopenicillinase (by cleavage of the membrane-bound enzyme) may well be dependent on the action of the releasing enzyme.

Pflugers Arch, 1976 Dec 28, 367(2), 177 - 81
The effect of an inhibitor of adenylate cyclase on the development of pyrogen, prostaglandin and cyclic AMP fevers in the rabbit; Willies GH et al.; An exotoxin of Bacillus thuringiensis known to inhibit adenylate cyclase in vitro has been used to investigate the role of cyclic AMP in the pathogenesis of fever in the rabbit . Intra-hypothalamic microinjections of the exotoxin are non-pyrogenic and significantly attenuate the hyperthermia caused by intrahypothalamic microinjections of both bacterial pyrogen (endotoxin) and prostaglandin E1 . The hyperthermia produced by dibutyrl cyclic AMP is not affected by the exotoxin . These results support the idea that adenylate cyclase is activated during the development of fever in the rabbit.

J Biol Chem, 1976 Dec 25, 251(24), 7853 - 62
Purification and properties of a specific proteolytic enzyme present in spores of Bacillus magaterium; Setlow P; A proteolytic enzyme with high activity on the specific, low molecular weight dormant spore proteins (termed A and B proteins) degraded during spore germination has been purified approximately 1000-fold from Bacillus megaterium spores . This enzyme accounts for greater than 85% of the proteolytic activity toward the A and B proteins in crude spore extracts . The protease has a pH optimum of approximately 7.5, is inactivated by phenylmethylsulfonyl fluoride and EDTA (10 mM), and is inhibited approximately 70% by NaCl (1 M) . The enzyme is unstable and requires glycerol, divalent cations, and high enzyme concentrations for maximum stability . The protease shows a high degree of specificity for the A and B proteins, since high levels of enzyme catalyze no detectable bond cleavage on a variety of amide, ester, peptide, or other protein substrates . The enzyme is an endoprotease and digestion of the A and B proteins in vitro generates a number of peptide fragments at least one of which appears identical with a fragment isolated from lysates of spores carrying out hydrolysis of endogenous A and B proteins . In vitro, the peptide products can be rapidly degraded to amino acids by an aminopeptidase which has also been partially purified from spores . Although high levels of the protease are present in extracts of dormant spores, and of spores germinated for a few minutes, the enzyme is undetectable in log phase and early stationary phase cells . Furthermore, the protease disappears rapidly (t1/2 approximately 30 min) and completely (greater than 90%) as the process of germination proceeds.

Biochim Biophys Acta, 1976 Dec 20, 450(3), 311 - 21
Enzymes of phospholipid synthesis in Bacillus Calmette-Guerin induced rabbit alveolar macrophage . Characterization and localization of cytidine diphosphocholine phosphotransferase and monoacylphospholipid acyltransferase; Wang P et al.; The rabbit alveolar macrophage is capable of renewing its plasma membrane by at least two metabolic pathways . It contains (1) a monoacylphospholipid acyltransferase, which catalyzes the synthesis of diacylphospholipids by recycling monoacylphospholipids produced by the action of phospholipases and (2) a cytidine diphosphocholine phosphotransferase (CDPcholine phosphotransferase), which catalyzes the last step in the synthesis de novo of diacylglycerophosphocholine . These activities have been characterized in the cell homogenate with respect to time, protein, pH optimum (for CDPcholine phosphotransferase), substrate specificity (for monoacylphospholipid acyltransferase) and cation requirement ( for CDPcholine phosphotransferase) . Monoacylphospholipid acyltransferase activity is localized solely in the endoplasmic reticulum . On the other hand, the CDPcholine phosphotransferase activity can be measured in the endoplasmic reticulum and in the plasma membrane, characterized by both differential and gradient sedimentation techniques . In addition to the normal route of phospholipid synthesis in the endoplasmic reticulum, the rabbit alveolar macrophage may thus possess the capacity for in situ synthesis of phospholipids of plasma membrane as a mechanism for membrane renewal following phagocytosis.

Biochim Biophys Acta, 1976 Dec 8, 452(2), 580 - 96
Dihydrodipicolinic acid synthase of Bacillus licheniformis . Quaternary structure, kinetics, and stability in the presence of sodium chloride and substrates; Halling SM et al.; Dihydrodipicolinic acid synthase (L-aspartate-beta-semialdehyde hydro-lyase (adding pyruvate and cyclising), EC 4.2.1.52) obtained from Bacillus licheniformis was purified to homogeneity . Its molecular weight was 108 000 to 117 500, depending on the concentration of NaCl and substrates present, and it contained four subunits of identical molecular weight (28000) . The Km values for pyruvate and L-aspartic semialdehyde were approximately 5.3 Km values for pyruvate and L-aspartic semialdehyde were approximately 5.3 and 2.6 mM, respectively . It was previously shown that pyruvate and a high sodium chloride concentration contributed to the stability of the enzyme . The effect of these substances and the other substrate, L-aspartic semialdehyde, on molecular weight was determined . None of these three substances significantly affected the apparent molecular weight . The effect of sodium chloride, pyruvate, and L-aspartic semialdehyde on enzyme structure was studied by determining the effect of their presence on inactivation of the enzyme by several chemical denaturants and heat . Pyruvate dramatically protected against inactivation by all of the denaturants . Sodium chloride protected against inactivation by sodium dodecyl sulfate, guanidine-HCl, urea, and heat, but somewhat facilitated inactivation by ethanol . L-Aspartic semialdehyde had no significant effect on inactivation by sodium dodecyl sulfate and ethanol; it rendered the enzyme slightly more sensitive to inactivation by guanidine-HCl and urea . The thermal melting curve obtained for the enzyme in the presence of L-aspartic semialdehyde was biphasic . The activity was reduced approximately 50% by heating for 30 min at temperatures between 50 and 80 degrees C . Only by heating at temperatures above 80 degrees C did the inactivation become complete . The partially inactivated enzyme could be reactivated by heating after removal of the L-aspartic semialdehyde . Pyruvate prevented the partial inactivation and facilitated reactivation . The only difference detected between the native enzyme and the partially inactivated form of the enzyme was that the latter had a reduced V . It is known that in other spore-formers, dihydrodipicolinate synthase increases in activity late in sporulation . This increase may be important for normal sporulation to occur . The possibility is discussed that the intracellular pool sizes of pyruvate and L-aspartic semialdehyde might have an influence on the level of dihydrodipicolinate synthase activity, by controlling the amount of partial inactivation of the enzyme that occurs in vivo.

Arch Biol Med Exp (Santiago), 1976 Dec, 10(1-3), 61 - 4
In vitro degradation of ribosomes; Mora G et al.; The cytoplasmic ribosomes from Euglena gracilis var . bacillaris are found to be of two types taking into consideration their stability "in vitro" . In the group of unstable ribosomes the large subunit is degraded . The other group apparently does not suffer any degradation under the conditions described . However the RNAs extracted from both types of ribosomes are degraded during sucrose density gradients . The degradation of the largest RNA species has been reported previously, but no comment has been made about the stability of the ribosome itself.

Chir Ital, 1976 Dec, 28(6), 665 - 74
{Recent aspects of pulmonary tuberculosis in thoracic surgery}; Puchetti V et al.; Pulmonary tuberculosis--although appreciable reduced as regards number of cases--is still far from being wiped out and even today constitutes a real problem in the field of thoracico-pulmonary surgical pathology . Infection with Koch's bacillus can from this point of view arise basically according to two procedures: a) as superimposed element--unexpected guest--on a pulmonary disease of surgical interest, sometimes being mistaken for it (e.g . a tubercular inflammation of slow evolution mistaken for a cancer of the lung) or b) taking the form of pulmonary relics of an earlier and apparently extinguished Kochian infection, the starting point of an always possible revival of the infection during the postoperative period . In the former case the so-called attack therapy must be adopted without delay in accordance with well tested doses and schedules; in the latter case by means of a chemoprophylaxis, carried out in with rifampicin (600/1200 mg/day) in the cases reported and also continued in the postoperative period.

J Biochem (Tokyo), 1976 Dec, 80(6), 1457 - 60
The site-specific deoxyribonuclease from Bacillus pumilus (endonuclease R.Bpu1387); Ikawa S et al.; A new site-specific endonuclease (DNase) was isolated from the cells of Bacillus pumilus AHU 1387 strain . This enzyme (endonuclease R.Bpu 1387) introduced double-stranded scissions at unique sites on DNA's of coli phage lambda, lambdadvl, coli phage T7, Bacillus phage phi105C, Bacillus phage SP10, and Simian Virus 40, in the presence of magnesium ion . The activity was stimulated by the presence of NaCl.

Br J Exp Pathol, 1976 Dec, 57(6), 706 - 12
Skin reaction and antibody responses in guinea-pigs sensitized to human leukaemia cells or their nuclei in combination with Bacillus Calmette-Guérin; Barnes RM et al.; Guinea-pigs sensitized by subcutaneous injection of chronic lymphatic leukaemia (CLL) cells combined with Bacillus Calmette-Guerin (BCG) displayed good skin reacitons 24 and 48 h after challenge with CLL cells . Equally good responses were also demonstrated using nuclei from the leukaemic cells in combination with BCG . These reactions were significantly greater than those produced in the same manner but without BCG . Sera form the animals were examined for the presence of antibodies against CLL cells by cytotoxicity and immunofluorescence techniques . Only samples from guinea-pigs innoculated with CLL cells were found to contain significant antibodies . Histological examination showed that whereas leukaemic cells persisted at the sensitizing injection site leukaemic cell nuclei could not be visualized . It is suggested that because leukaemic cell nuclei in combination with BCG are able to induce good skin reactivity without provoking a vigorous humoral antibody response they may have possible advantages over leukaemic cells when used for immunotherapy.

J Antibiot (Tokyo), 1976 Dec, 29(12), 1281 - 5
Production and isolation of cerexins C and D studies on antibiotics from the genus Bacillus . XVII; Shoji J et al.; From the culture broth of Bacillus cereus 60-6, in which antibiotic production was improved a new antibiotic, named cerexin C, was isolated . Similarly, a new antibiotic, cerexin D was isolated from the culture broth of Bacillus cereus Gp-3 . Cerexins C and D are closely related to cerexins A and B in their physico-chemical and antimicrobial properties . In fact cerexins C and D are peptides essentially identical with cerexins A and B except for the presence of a lysine residue in place of the gamma-hydroxylysine residue.

J Antibiot (Tokyo), 1976 Dec, 29(12), 1275 - 80
The structure of cerexin B (studies on antibiotics from the genus Bacillus . XVII); Shoji J et al.; The structure of cerexin B was examined . The constitutent fatty acids were elucidated by gas chromatography and mass spectrometry to be beta-hydroxy isodecanoic acid, beta-hydroxy decanoic acid, beta-hydroxy isoundecanoic acid and beta-hydroxy anteisoundecanoic acid . The configurations of constituent amino acids were determined as asparagine (2D, 1L), valine (D), phenylalanine (D), allo-threonine (D), tryptophan (D), and allo-isoleucine (D) from their optical activities . Treatment with conc . hydrochloric acid cleaved at the gamma-hydroxylysine residue to give two peptide fragments, one of which (the N-terminal side) was then deacylated with Polymyxin Acylase . Their amino acid sequences were examined by EDMAN degradation . From the results and analogy to cerexin A, the structure of cerexin B was deduced.

J Antibiot (Tokyo), 1976 Dec, 29(12), 1268 - 74
The total structure of cerexin A (studies on antibiotics from the genus Bacillus . XVI; Shoji J et al.; The constituent fatty acid of cerexin A was elucidated to be beta-hydroxy isoundecanoic acid by gas chromatography, mass spectrometry and nuclear magnetic resonance . The asparaginyl asparagine linkage in the amino acid sequence of the antibiotic, which has been previously described, was proved to be a normal alpha-carboxylpeptide bond by dehydration-reduction procedure . The three asparagine residues in the sequence were isolated separately and their chiralities were determined . The structure of cerexin A was concluded from the above results.

J Gen Microbiol, 1976 Dec, 97(2), 289 - 96
Immunofluorescence detection of nitrogenase proteins in whole cells; Rennie RJ; Fluorescent antibodies (FA) prepared against the Mo-Fe and Fe proteins of nitrogenase from Klebsiella pneumoniae M5aI were used to detect these protein components in toluene-treated whole cells that were actively reducing acetylene . The FA were highly specific, staining only nitrogenase component proteins originating from Klebsiella . Cross-reactions between the FA and purified nitrogenase proteins from other dinitrogen-fixing micro-organisms did not occur, except in the case of Bacillus polymyxa . The tests rapidly and accurately assayed the component proteins in Klebsiella mutants and derivatives to which Klebsiella nif genes had been transferred either by plasmid or by other means . Cross-reactions also indicated the degree of relatedness between nitrogenase proteins from dinitrogen-fixing micro-organisms of various origins.

Cancer Res, 1976 Dec, 36(12), 4459 - 66
A comparison of in vitro cell-mediated reactivity against syngeneic tumor cells by various lymphoid cell populations from Bacillus Calmette-Guérin-tumor-cured, tumor-sensitized, tumor-bearing, and normal inbred guinea pigs; Fidler IJ et al.; The cell-mediated reactivity (CMR) in vitro of normal, line 10 hepatocardinoma tumor-bearing, Bacillus Calmette Guerin (BCG)-line 10 tumor-cured, and line 1 sensitized guinea pigs against synegeneic line 10 hepatocarcinma was measured . This study was designed to determine whether the in vitro CMR correlates with the clinical stage of disease in a guinea pig immunotherapy model . A comparison was also made between the CMR of effector cells from active regional lymph nodes, pooled distant lymph nodes and spleen non-glass-adherent mononuclear cells (lymphocytes), peritoneal exudate macrophages (PEM), and peripheral blood leukocytes . In addition, the immunological specificity of the in vitro cytotoxicity was investigated by comparing the reactivities against line 10 and the highly antigenic syngeneic line 1 hepatocarcinoma as well as normal syngeneic guinea pig embryo cells . Pooled lymphocytes and PEM from normal guinea pigs were cytotoxic to line 1 hepatocarcinoma cells but not to line 10 hepatocarcinoma cells . Pooled lymphocytes and PEM obtained from animals sensitized in vivo to line 1 tumor exhibited in vitro cytotoxicity against line 1 cells, which was comparable to that cytotoxicity obtained from normal, naive donors . Various effector cells from BCG-tumor-cured guinea pigs destroyed line 10 cells but to different degrees . In the order of their decreasing cytotoxic effect against line 10 cells, the effector cells from BCG-tumor-cured guinea pigs were: glass-adherent regional lymph node cells and PEM, followed by non-glass-adherent regional lymph node cells, peripheral blood leukocytes, and pooled spleen and lymph node lymphocytes . Effector cells obtained from animals bearing line 10 tumors were all cytotoxic in vitro against line 10 cells . There were no qualitative differences in CMR between effector cells of tumor-bearing and BCG-tumor-cured guinea pigs . Although some quantitative differences between tumor-bearing and tumor-cured guinea pigs were observed, these differences were not large enough to reliably predict the clinical status of the donor in a blind test.

Cancer Res, 1976 Dec, 36(12), 4444 - 58
Morphological evidence for the translocation of lysosomal organelles from cytotoxic macrophages into the cytoplasm of tumor target cells; Bucana C et al.; In this study we have recorded in detail the morphological sequence of interactions between activated macrophages and tumor target cells in vitro . The study is unique because it involves the combination of several microscopic techniques that are used sequentially to study a single cell-to-cell interaction . Many such cellular interactions were examined first by time lapse cinematography; then the effector cell was identified by specific immunofluorescence, and the areas of interaction were processed for scanning and then transmission electron microscopy . The tumor target cells were guinea pig line 10 hepatocarcinoma cells . The effectors were peritoneal exudate cells (PEC) harvested from syngeneic strain 2 guinea pigs that had been cured of a line 10 tumor by intratumoral injection of Bacillus Calmette-Guerin . Host-target cell interactions between (a) line 10 cells and PEC from Bacillus Calmette Guerin-tumor-cured animals, (b) line 10 cells and PEC from normal animals and (c) syngeneic guinea pig embryo cells and PEC from Bacillus Calmette-Guerin-tumor-cured animals were studied . These comparisons demonstrate that the mechanism of tumor cell killing by activated macrophages is a nonphagocytic process . Our results suggested that the macrophage-tumor cell interaction is initiated by a recognition phase that results in extracellular release of lysosomes through macrophage exocytosis and clasmatosis . Neoplastic target cell susceptibility may be the result of an active or passive uptake of lysosomes and consequently cytolysis.

Jpn J Microbiol, 1976 Dec, 20(6), 529 - 35
Germination of unactivated spores of Bacillus cereus T . Effect of preincubation with L-alanine or inosine on the subsequent germination; Shibata H et al.; Heat-activated spores of Bacillus cereus T germinate rapidly in the presence of L-alanine alone or inosine alone . In contrast, unactivated spores can not germinate in the presence of either germinant alone but rapidly in the presence of both germinants . The highest level of cooperative action of L-alanine and inosine on the germination was observed when they were present in a ratio 1:1 . Preincubations of unactivated spores with L-alanine or inosine had opposite effects on the subsequent germination in the presence of both germinants: preincubation with L-alanine stimulated the initiation of subsequent germination, while preincubation with inosine inhibited it . These results suggest that germination of unactivated spores initiated by L-alanine and inosine includes two steps, the first initiated by L-alanine and the second prompted by inosine . The effect of preincubation of unactivated spores with L-alanine was not diminished by washings . The pH dependence of the preincubation of unactivated spores was not so marked as that of the subsequent germination in the presence of inosine.

Can J Microbiol, 1976 Dec, 22(12), 1743 - 50
Isolation and characterization of a bacteriocin produced by Bacillus stearothermophilus strain NU-10; Yule R et al.; Broth-grown cultures of Bacillus stearothermophilus strain NU-10 produce a bacteriocin which exerts lethal activity on other strains of the bacterium . Optimal production occurs during late maximum stationary phase of growth, at neutral pH, and 55-65 degrees C . The bacteriocin can be substantially purified by a combination of precipitations, centrifugations, and gel filtrations . The thermocin is composed of protein and carbohydrate . It is partially destroyed by proteolytic enzymes but is resistant to DNase, RNase, and various chemical treatments . The bacteriocin has a small molecular weight and exhibits considerable thermostability.

Science, 1976 Nov 26, 194(4268), 954 - 6
Bacillus thuringiensis enzyme-digested delta endotoxin: effect on cultured insect cells; Murphy DW et al.; Cells from three insect cell lines responded to the enzyme-digested delta endotoxin of Bacillus thuringiensis with swelling, lysis, and vesicle formation . Sufficient toxin was taken up in 1 minute to cause half-maximal cell damage . Cytoxic activity was neutralized by specific antiserum to the endotoxin.

C R Acad Sci Hebd Seances Acad Sci D, 1976 Nov 22, 283(13), 1563 - 6
{Variations of enzymatic activities in an asporogenic mutant of Bacillus megaterium for various nutritional deficiencies}; Chatelain S et al.; An asporogenous mutant of Bacillus megaterium KM, blocked at stage I of sporulation, is cultivated in different conditions which usually lead to sporulation (Nitrogen, carbon, phosphate starvation) . Aklaline phosphatase appears only in inorganic phosphate starvation like the wild type, but a glucose dehydrogenase activity also appears in phosphate starvation and so, is dissociated from sporulation.

Biochim Biophys Acta, 1976 Nov 19, 450(2), 154 - 64
Studies on phosphatidylinositol phosphodiesterase (phospholipase C type) of Bacillus cereus . I . purification, properties and phosphatase-releasing activity; Ikezawa H et al.; A phosphatidylinositol phosphodiesterase from the culture broth of Bacillus cereus, was purified to a homogeneous state as indicated by polyacrylamide gel electrophoresis, by ammonium sulfate precipitation and chromatography with DEAE-cellulose and CM-Sephadex . The enzyme (molecular weight: 29000 +/- 1000) was maximally active at pH 7.2-7.5, AND NOT INFLUENCED BY EDTA, ophenanthroline, monoiodoacetate, p-chloromercuribenzoate or reduced glutathione . The enzyme specifically hydrolyzed phosphatidylinositol, but did not act on phosphatidylcholine, phosphatidylethanolamine and sphingomyelin, under the conditions examined . The products from phosphatidylinositol of enzyme reaction were diacylglycerols and a mixture of myoinositol 1- and 1, 2-cyclic phosphates, suggesting that the enzyme was a phosphatidylinositol-specific phospholipase C . The enzyme released alkaline phosphatase quantitatively from rat kidney slices . A kinetic analysis was made on the release of alkaline phosphatase . The results suggest that phosphatidylinositol-specific phospholipase C can specifically act on plasma membrane of rat kidney slices.

Experientia, 1976 Nov 15, 32(11), 1465 - 6
The exotoxin of Bacillus thuringiensis: a new C-mitotic agent; Sharma CB et al.; Thuringiensin A, an exotoxin from Bacillus thuringiensis, a constituent of the microbial insectide thuricide has been found to inhibit mitotic spindle, condense and scatter chromosomes . It may therefore be a promizing tool in future cell biological studies.

Experientia, 1976 Nov 15, 32(11), 1373 - 4
The citrate synthase from Bacillus Stearothermophilus; Higa AI et al.; Citrate synthase has been purified to apparent homogeneity from Bacillus stearothermophilus . Its kinetic and regulatory properties, and molecular weight, are similar to those of the enzymes from suitable mesophilic counterparts, but its thermal stability is considerably greater.

JAMA, 1976 Nov 8, 236(19), 2187 - 9
Skeletal and soft tissue sarcomas . Treatment with adjuvant immunotherapy; Townsend CM Jr et al.; Because there is evidence for an active immunologic response against sarcoma, a clinical trial of adjuvant immunotherapy using bacillus Calmette-Guerin (BCG) and tumor cell vaccine was begun . Eleven of 18 patients with localized soft tissue sarcoma who received immunotherapy are free of disease, compared to only 5 of 15 treated by operation alone who are free of disease . Furthermore, immunotherapy also prolonged the median disease-free interval from 7.3 months to 15 months in the patients who experienced recurrence of their disease.

Arch Microbiol, 1976 Nov 2, 110(23), 279 - 86
Toxicity and accumulation of thallium in bacteria and yeast; Norris P et al.; Thallium sulphate inhibited microbial growth, with Bacillus megaterium KM more sensitive to the metal than Saccharomyces cerevisiae and Escherichia coli . Inhibition of B . megaterium KM and S . cerevisiae, but not of E . coli, was alleviated by increasing the potassium concentration of the medium; inhibition of respiration of S . cerevisiae, but not of E . coli, was similarly alleviated . Thallium was rapidly bound, presumably to cell surfaces, by S . cerevisiae and E . coli, and was progressively accumulated by energy-dependent transport systems (probably concerned primarily with potassium uptake) with both organisms . Thallium uptake kinetics suggested more than one transport system operated in yeast, possibly reflecting a multiplicity of potassium transport systems . Apparent Km and Ki values for competitive inhibition of thallium uptake by potassium indicated S . cerevisiae to have a higher affinity for thallium uptake than for potassium, while E . coli had a transport system with a higher affinity for potassium than for thallium . The likely systems for thallium transport are discussed . A mutant of E . coli with tenfold decreased sensitivity to thallium was isolated and apparently effected surface binding of thallium in amounts equivalent to the wild type organism, but showed no subsequent uptake and accumulation of the metal from buffer, even though it was able to accumulate potassium to normal intracellular concentrations during growth.

Parazitologiia, 1976 Nov-Dec, 10(6), 552 - 4
{Effect of the bacterial preparation, insectin, on the fleas Xenopsylla cheopis}; Ershova LS et al.; The effect of 0.2-2 mg/g of insectine (Bacillus insectus) upon the Ist-IInd stage larvae of X . cheopis is very negligible but it manifests itself in the subsequent low (9 to 30%) development and hatching of imagos . The III stage larvae are more resistent and after the effect of 10 mg/g of the preparation 9% of larvae formed cocoons which later developed into imagos . Adult fleas poorly responded to 0.2-2 mg/g of insectine . However, 10-30 mg/g of insectine caused in four days a mass death of fleas and their population decreased to 70% as compared to control.

Mikrobiologiia, 1976 Nov-Dec, 45(6), 1075 - 81
{Microbiological studies of the Wanda Lake (Antarctica)}; Kriss AE et al.; The deep-water (68 m) Wanda Lake in the Antarctica is noticeable by three characteristics: a relatively high temperature in the bottom layer, elevated salinity, and the presence of H2S . Only several (less often, dozens) saprophytic microorganisms per 40 ml of water are encountered in the lake . The total number of bacteria varies from 6-10(3) to 172-10(3) cells/ml . The highest content of the total bacterial population, and saprophytes, is found in intermediate layers, 30 and 40-50 m deep, respectively . Microbial strains isolated from water and ooze belong to the genera Pseudomonas, Chromobacterium, Bacillus, and Mycobacterium . Yeast organisms were also found . Sulphate reducing bacteria were detected only at one station in ooze of the lake while thionic bacteria could not be determined at all . Photosynthetic bacteria were isolated from ooze at all four stations.

J Infect Dis, 1976 Nov, 134 SUPPL, S286 - 90
Amikacin in treatment of infections caused by gram-negative bacteria resistant to gentamicin and other aminoglycosides: clinical and bacteriologic results; Daikos GK et al.; Amikacin (250 or 500 mg) was administered intramuscularly twice daily at 12-hr intervals to 34 patients with infections due to various gram-negative bacteria . Usually one or more aggravating factors were present, such as serious underlying pathology or therapy with steroids or immunosuppressants . Clinical isolates from most patients were resistant to gentamicin and other aminoglycosides . The overall response to therapy was excellent in 20 patients; in eight patients clinical response was good, but the organism persisted . Six patients showed some improvement without complete resolution of the infection or eradication of the causative organism . There were no complete clinical or bacteriologic failures . Ototoxicity was not observed in any patient . Levels of blood urea nitrogen and serum creatinine increased in two patients but returned to pretreatment levels within two weeks after therapy . No other adverse reactions were noted . Amikacin may replace gentamicin as initial therapy in serious gram-negative bacillary infections, particularly when resistance to gentamicin is a problem.

Int J Radiat Biol Relat Stud Phys Chem Med, 1976 Nov, 30(5), 419 - 32
Effects of some -OH scavengers on the radiation sensitization of bacterial spores by p-nitroacetophenone and O2 in suspension; Ewing D; Spores of Bacillus megaterium were irradiated in suspension with 50 kVp X-rays under three reference conditions: in anoxia (i.e., 100 per cent N2); in anoxia with 2mM p-nitroacetophenone (PNAP), a concentration that shows the maximum amount of sensitization by this agent; and in air . The responses were then measured when different concentrations of an hydroxyl radical scavenger were also present . Allyl alcohol, t-amyl alcohol, t-butanol, ethanol, glycerol, the formate ion, and methanol were the -OH scavengers that were used . A comparison of the effects these additives have on the three reference responses provides indirect, comparative information on the sensitizing processes of PNAP and O2 . Neither t-amyl alcohol nor t-butanol affects the response in air or N2 alone . Both these additives, however, can reduce the sensitization from PNAP slightly . The other additives have significant protective effects under the three conditions . In general, the additives have similar effects in air and in 100 per cent N2 but much greater protective effects in PNAP . Although the sensitizing mechanisms are not identified by these experiments, the effects of O2 and PNAP, as judged by the desensitizing actions of these additives, appear quite different.

Proc Natl Acad Sci U S A, 1976 Nov, 73(11), 4200 - 4
Oligomeric intermediate in peptidoglycan biosynthesis in Bacillus megaterium; Fuchs-Cleveland E et al.; An oligomeric intermediate in the biosynthetic pathway of peptidoglycan was isolated from B . megaterium . The oligomer has been identified as {disaccharide(pentapeptide)}12 pyrophosphoryl undecaprenol.

Cancer Lett, 1976 Nov, 2(2), 93 - 100
Influence of five different postnatal lifelong treatments on the transplacental carcinogenicity of ethylnitrosourea in Sprague-Dawley rats; Habs M et al.; Sprague-Dawley rats received in 10 mg/kg body weight (group I) or 30 mg/kg body weight (group II) ethylnitrosourea (ENU) orally on the 19th day of pregnancy . Their offspring were treated with Bacille Calmette-Guerin, human albumin, hydrocortisone, cyclophosphamide or nicotine starting on the 6th day of life . The ENU treatment significantly reduced the life expectancy of all offspring . Treatment of the offspring did not influence tumor frequency, induction time or localization of neurogenic malignant tumors . Cyclophosphamide treatment of group II offspring increased the number of females bearing mammary carcinomas.

Mikrobiologiia, 1976 Nov-Dec, 45(6), 966 - 72
{Chitinase of Bacillus thuringiensis}; Chigaleichik AG; Strains of Bacillus thuringiensis were shown to hydrolyse various forms of chitin around growing colonies on a solid medium . In conditions of submerged cultivation on a medium containing demineralized crab shells, Bac . thuringiensis var . caucasicus INMI Arm . 837 manifests the chitinolytic activity at the beginning of the stationary growth phase . The activity of chitinase which is of a constitutive nature increases when the bacterium is cultivated at pH 7.2 . The maximum rate of hydrolysis of colloid chitin by chitinase prepared from the cultural broth is displayed at pH 8.0 and 60 degrees C.

J Bacteriol, 1976 Nov, 128(2), 651 - 7
Subunits of the alkaline phosphatase of Bacillus licheniformis: chemical, physicochemical, and dissociation studies; Hulett FM et al.; The alkaline phosphatase (orthophosphoric monoester phosphydrolase, EC 3.1.3.1) of Bacillus licheniformis MC14 was studied in an attempt to determine the number of subunits contained in the 120,000-molecular-weight native enzyme . Two moles of arginine was liberated per mole of native enzyme by carboxypeptidases A and B in the presence of sodium dodecyl sulfate . The effect on the native enzyme of progressively lowering the solvent buffer pH was monitored by determining the molecular weight by sedimentation equilibrium analysis, the sedimentation coefficient, the frictional coefficient, and the percent alpha-helix content of the enzyme . The alkaline phosphatase dissociates into two subunits around pH 4 . At pH 2.8 a further decrease in S value, but no change in molecular weight, is observed, indicating a change in conformation . The frictional coefficients and percent alpha-helix content agree with this interpretation . A subunit molecular weight of 59,000 was calculated from sodium dodecyl sulfate gels.

J Bacteriol, 1976 Nov, 128(2), 515 - 21
Stable, inducible thermoacidophilic alpha-amylase from Bacillus acidocaldarius; Buonocore V et al.; Bacillus acidocaldarius Agnano 101 produces an inducible thermoacidophilic alpha-amylase . The enzyme production occurs during the stationary phase of growth in the presence of compounds with alpha-1,4-glucosidic linkages . The enzymatic activity is both present in the culture medium and associated with the cells; the enzymes purified from both sources show identical molecular and catalytic properties . The purified amylase has a single polypeptide chain of molecular weight 68,000 and behaves like an alpha-amylase with affinity constants for starch and related substances of 0.8 to 0.9 mg/ml . The pH and temperature optima for activity are 3.5 and 75degreesC, respectively . The amylase is stable at acidic pH (below 4.5) . Its thermal stability is strictly dependent upon protein concentration; the half-life at 60degreesC of the amylase in a 70-mug/ml solution is about 5 days.

Biochim Biophys Acta, 1976 Oct 19, 448(3), 401 - 10
Membrane associated phospholipoproteins of Bacillus lichenformis 749; Aiyappa PS et al.; The membrane-bound penicillinase of Bacillus licheniformis 749/C is a phospholipoprotein that differs from the hydrophilic exoenzyme in that its polypeptide chain carries an additional 25 residues (mostly hydrophilic) with phosphatidylserine as the NH2-terminus . To determine if other phospholipoproteins are present in the plasma membrane, the penicillinase-inducible strain 749 was grown without inducer in the presence of {2-(3)H} glycerol . Electrophoretic separation of the membrane proteins (after removal of free lipids) showed an association of 3H-activity with certain of the proteins which could not be broken by lipid solvents and strongly denaturing conditions . Pronase digestion of the membrane proteins (after solvent extraction) released phosphatidylserine, thus indicating the covalent linkage of protein and phospholipid . Treatment of the isolated membranes with trypsin solubilized the protein portion of some of the phospholipoproteins (as with penicillinase), but not the 3H-labelled fragment . Penicillinase should be considered as the first observed example of a group of phosphatidylserine-containing proteins present in the plasma membrane of B . licheniformis 749 and 749/C.

J Biol Chem, 1976 Oct 10, 251(19), 6154 - 6
Identification of an isolated protein essential for peptidoglycan synthesis as the N-acetylglucosaminyltransferase; Taku A et al.; We have identified a protein extractable from Bacillus megaterium membranes by LiCl as the enzyme in peptidoglycan synthesis which catalyzes the transfer of N-acetylglucosamine from UDP-N-acetylglucosamine to N-acetyl-D-muramyl-L-analyl-D-gamma-glutamyl-meso-diaminopimelyl-D-alanyl-D-alanine-P-P-lipid . This identification demonstrates the isolation and characterization of a membrane-associated enzyme involved in a complex series of reactions without the ready availability of a substrate for the enzyme.

Biochemistry, 1976 Oct 5, 15(20), 4399 - 403
Purification and properties of guanosine 5', 3'-polyphosphate synthetase from Bacillus brevis; Sy J et al.; A ribosome-independent guanosine 5',3'-polyphosphate synthetase has been highly purified from Bacillus brevis (ATCC 8185) . The enzyme has a molecular weight of 55,000, as measured by sucrose density gradient centrifugation . Like the ribosome-connected stringent factor of Escherichia coli, it catalyzes the synthesis of the guanosine 5', 3'-polyphosphates by a pyrophosphoryl transfer mechanism from adenosine triphosphate (ATP) to guanosine di- or triphosphates (GDP, GTP) . It has an apparent Km of 0.14 mM for GDP and 0.77 mM for GTP, and is specific for the guanosine ribonucleotides as pyrophosphoryl acceptors . Several ATP analogues were tested for their ability to donate the pyrophosphoryl group . Mg2+ was required as a counter ion for the nucleotide substrate; however, an excess of Mg2+ was inhibitory . The property of the B . brevis enzyme is compared with the ribosome-linked enzyme of E . coli and an extracellular enzyme excreted by several types of Streptomyces reported upon recently.

Rev Rhum Mal Osteoartic, 1976 Oct, 43(10), 577 - 82
{Dysenteric rheumatism . What remains in 1976?}; Doury P et al.; Joint manifestations that occur during or after an episode of dysentery, grouped under the name of dysenteric rheumatism, have been known since antiquity, and have been mainly found in military circles, in camps, or during military campaigns . After reviewing the principal epidemics of rheumatism complicated by joint manifestations, the authors show that the only different feature about what is called dysenteric rheumatism is the particular character of the joint manifestations which may be isolated or associated with other manifestations, notably ocular or urethral, forming then what is called the Fiessinger-Leroy-Reiter syndrome . The role of the dysentery bacillus in the genesis of the joint lesions is not clear . Although it may been suspected on the basis of epidemiological or bacteriological arguments in some cases, it has never been formally demonstrated and is often non-existent . The term dysenteric rheumatism is too restrictive and should henceforth be abandoned.

J Biochem (Tokyo), 1976 Oct, 80(4), 867 - 74
Metabolism of threo-beta-methylmalate by a soil bacterium; Suzuki S et al.; Studies on threo-beta-methylmalate metabolism in a soil bacterium of the genus Bacillus which can utilize threo-beta-methylmalate as a sole carbon source were carried out . When DL-threo-beta-methylmalate was incubated with a cell-free extract of the bacterium, citramalate was found to be formed . Similarly, formation of threo-beta-methylmalate from DL-citramalate was confirmed . These dicarbosylic acids were identified by gas chromatography-mass spectrometry . Examination of inducibility, substrate specificity, and cofactor requirement of the enzymes involved in the reactions showed the existence of two interconversion reactions between the threo-beta-methylmalate and citramalate . One was an interconversion reaction between L-threo-beta-methylmalate and L-citramalate via mesaconate and the other was an interconversion reaction between D-threo-beta-methylmalate and D-citramalate via citraconate . These reactions were both reversible and were catalyzed by distinct and inducible enzymes . It is suggested that the two reactions participate in the catabolism of threo-beta-methylmalate.

Scand J Clin Lab Invest, 1976 Oct, 36(6), 553 - 9
Parenteral administration of phospholipase C in the rat . Distribution, elimination, and lethal doses; Otnaess AB et al.; Infusions of tissue thromboplastin induce intravascular coagulation in animals . Phospholipase C (PLC) (EC 3.1.4.3) from Bacillus cereus has a marked protective effect against such infusions and might be of value in the therapy of certain types of intravascular coagulation . We have therefore studied the toxicity, half-life, and effect on lipolysis of purified PLC in rats, using parenteral administration of 14C-labelled enzyme . Following intravenous injection, the plasma half-life was 5.2-5.4 min, and LD50 was approximately 1.6-1.7 mg/kg . The effect of PLC on lipolysis was moderate . The enzyme does not appear to be bound to any plasma macromolecules, and there was no accumulation of labelled enzyme in tissues other than kidney.

Biochem J, 1976 Oct 1, 159(1), 55 - 63
Amino acid sequence of a four-iron-four-sulphur ferredoxin isolated from Bacillus stearothermophilus; Hase T et al.; 1 . The primary structure of a 4Fe-4S ferredoxin from Bacillus stearothermophilus was determined and shown to consist of a single polypeptide chain of 81 amino acid residues . The molecular weight of the holoprotein is about 9120 . 2 . There are only four cysteine residues in the molecule; three of these are located near the N-terminus as a Cys-X-X-Cys-X-X-Cys segment, and the fourth cysteine residue is followed by a proline and located in the C-terminal half . 3 . The Fe-S chromophore in B . stearothermophilus ferredoxin was previously well characterized and was shown to consist of a single 4Fe-4S cluster . This ferredoxin sequence establishes for the first time the relative location of the four cysteine residues necessary to bind the 4Fe-4S cluster of a 4Fe ferredoxin, and is in agreement with the criteria for the relative positions of the cysteines proposed from X-ray-crystallographic studies on an 8Fe (two 4Fe-4S clusters) ferredoxin . 4 . The sequence of B . stearothermophilus ferredoxin is homologous in many segments to that of other bacterial ferredoxins, the degree of homology being greater towards ferredoxins from Desulfovibrio gigas and photosynthetic bacteria than to Clostridial ferredoxins . 5 . The presence of a relatively higher number of glutamic acid and lower number of cysteine residues in the molecule may explain the greater thermal stability and oxygen-insenstivity of this ferredoxin.

Biochem J, 1976 Oct 1, 159(1), 121 - 32
Subsite mapping of enzymes . Application of the depolymerase computer model to two alpha-amylases; Allen JD et al.; In the preceding paper (Allen and Thoma, 1976) we developed a depolymerase computer model, which uses a minimization routine to establish a subsite map for a depolymerase . In the present paper we show how the model is applied to experimental data for two alpha-amylases . Michaelis parameters and bond-cleavage frequencies for substrates of chain lengths up to twelve glucosyl units have been reported for Bacillus amyloliquefaciens, and a subsite map has been proposed for this enzyme {Thoma et al . (1971) J . Biol . Chem . 246, 5621-5635} . By applying the computer model to the experimental data, we have arrived at a ten-subsite map . We find that a significant improvement in this map is achieved by allowing the hydrolytic rate coefficient to vary as a function of the number of occupied subsites comprising the enzyme-binding region . The bond-cleavage frequencies, the enzyme is found to have eight subsites . A partial subsite map is arrived at, but the entire binding region cannot be mapped because Michaelis parameters are complicated by transglycosylation reactions . The hydrolytic rate coefficients for this enzyme are not constant.

Infect Immun, 1976 Oct, 14(4), 934 - 41
Evidence for two immune inhibitors from Bacillus thuringiensis interfering with the humoral defense system of saturniid pupae; Edlund T et al.; Mutants of Bacillus thuringiensis lacking either beta-exotoxin or gamma-endotoxin were compared for their virulence using pupae of a giant silk moth . Known doses of viable log-phase bacteria were injected, and the response was followed as the number of viable bacteria in the hemolymph . The results obtained imply that, in the system used, neither the beta-exotoxin nor the gamma-endotoxin and the sporeforming ability are of importance for virulence . Results with sterile culture filtrate from B . thuringiensis have given evidence for the production of two inhibitors, A and B, which interfere with the humoral defense system in pupae of Hyalophora cecropia . Inhibitor A, which blocked the lysis of Escherichia coli,was precipitated by trichloroacetic acid and sensitive to heating . Inhibitor B, which blocked the killing of Bacillus cereus, was soluble in trichloroacetic acid and resistant to 90 degrees C for 5 min . Both inhibitors are believed to contribute to the insecticidal nature of B . thuringiensis.

Appl Environ Microbiol, 1976 Oct, 32(4), 508 - 10
Commercial formulations of Bacillus thuringiensis for control of Indian meal moth; Schesser JH; Doses of four commercial formulations and one experimental formulation of Bacillus thuringiensis Berliner were mixed with the diet used to rear colonies of the Indian meal moth Plodia interpunctella (Hubner) . Indian meal moth eggs were introduced to the treated diet, and the resultant adult emergence was tabulated . The experimental formulations ranked as follows in efficacy in controlling the Indian meal moth: Dipel (50% lethal concentration {LC50}, 25 mg/kg) greater than Bactospeine WP (LC50, 100 mg/kg) greater than Thuricide (LC50, 150 mg/kg) greater than IMC 90007 (LC30, 180 mg/kg) greater than Bactospeine Flowable (LC50, 440 mg/kg).

J Dairy Sci, 1976 Oct, 59(10), 1830 - 4
Fatty acids of vegetative cells and spores of Bacillus licheniformis; Martin JH; Lipids were extracted from vegetative cells and spores of Bacillus licheniformis . Vegetative cells were grown in nutrient broth and spores on nutrient agar . Total lipid approximated 2.89% of the dry weight of vegetative cells and 2.09% of the dry weight of spores . The fatty acids were prepared as methyl esters and analyzed by gas chromatography and mass spectrometry . There were six fatty acids in concentrations greater than 5% of the total lipid in both spores and vegetative cells, but only palmitic acid was common to both . Fatty acids from vegetative cells in quantities of 5% or more of the total lipid material were lauric, myristic, palmitic, palmitoleic, and linoleic acids . Fatty acids from spores in concentrations greater than 5% of the total lipid were isopentadecylic, palmitic, Carbon-17 iso, and three other long or branched chain fatty acids which were not identified . Spores contained more long and branched chain fatty acids with odd numbers of carbon atoms than did vegetative cells.

J Bacteriol, 1976 Oct, 128(1), 506 - 9
Light-induced inhibition of sporulation in Bacillus licheniformis; Propst-Ricciuti C et al.; Sporulation of Bacillus licheniformis is inhibited by broad-spectrum light . This phenomenon is intensity dependent and is a near-ultraviolet and blue light effect.

J Bacteriol, 1976 Oct, 128(1), 149 - 56
Teichoic acids and lipids associated with the membrane of a Bacillus licheniformis mutant and the membrane lipids of the parental strain; Button D et al.; Bacillus licheniformis 6346 MH-1 and a phosphoglucomutase-deficient poorly lytic mutant, B . licheniformis 6346 MH-5, both contain cardiolipin, phosphatidyl ethanolamine, and phosphatidyl glycerol but are devoid of phosphoglycolipids . Gentiobiosyl diglyceride is present in the parent organism but glycolipids are absent from the mutant . Lipoteichoic acid was extracted from the whole cells of MH-5 with hot aqueous phenol and contained fatty acids, glucosamine, and 1,3-polyglycerol phosphate . The fatty acids were predominantly of the branched-chain type and were esterified to hydroxyl groups of a terminal glycerol residue . The polyglycerol phosphate chains contained, on average, 32 to 40 glycerol residues, some of which were substituted at the secondary hydroxyl group with alpha-N-acylglucosaminyl residues . Phenol extraction of the supernatant fluid that remained when walls were removed from preparations of disrupted cells of MH-5 yielded membrane teichoic acid, which consisted of substituted polyglycerol phosphate but was devoid of fatty acids.

Scand J Haematol, 1976 Oct, 17(4), 305 - 11
Protection of rats by phospholipase C from Bacillus cereus against the effects of intravenous infusions of purified tissue thromboplastin; Giercksky KE et al.; Infusions of purified tissue thromboplastin in rats cause the accumulation of fibrin and platelets in the lungs and produce marked changes in the platelet count and in the coagulation factors V, VII and VIII . Tissue thromboplastin in a dose corresponding to less than 2 mug of protein per rat is lethal when given as a bolus injection . Simultaneous i.v . administration purified phospholipase C effectively prevents all these changes and protects rats from otherwise lethal doses of tissue thromboplastin . The necessary doses of phospholipase C are well below the toxic level for phospholipase C alone.

J Gen Microbiol, 1976 Oct, 96(2), 393 - 9
On the role of bacitracin peptides in trace metal transport by Bacillus licheniformis; Haavik HI; Bacitracin markedly increased the toxic effect of several divalent metal ions towards growth of the producer strain Bacillus licheniformis ATCCI4580 . Magnesium ions antagonized the toxic effect of these divalent cations both in the presence and absence of bacitracin . It is suggested that bacitracin increases the uptake of several divalent metal ions . The function of the bacitracin peptides may be to extract essential divalent cations from 'waiting sites' on the surface of the cells and transfer the cations to the transport mechanisms in the cytoplasmic membrane.

South Med J, 1976 Oct, 69(10), 1317 - 22
Atypical radiographic appearance and clinical presentation of pulmonary tuberculosis; Lian CF et al.; Fourteen patients with unusual presentation of pulmonary tuberculosis are described . Often, exposure to the tubercle bacillus was not apparent . In other cases, there was tuberculous infection or reactivation of a previous focus in individuals with altered immunity . The radiographic presentation often differed from the classic cavitating upper lobe disease . Persistent pneumonia, cavities, and nodules were seen in other areas of the lung including the anterior segments and the lung bases . Isolated effusion and solitary nodules were also evident . Increased suspicion of tuberculosis should be maintained when evaluating persistent parenchymal consolidation, effusion, and cavitating pulmonary disease.

Proc Natl Acad Sci U S A, 1976 Oct, 73(10), 3628 - 32
Evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions; Hozumi N et al.; A high-molecular-weight DNA from Balb/c mouse early embryo or from MOPC 321 plasmacytoma (a k-chain producer) was digested to completion with Bacillus amyloliquefaciens strain H restriction enzyme (BamH I) . The resulting DNA fragments were fractionated according to size in preparative agarose gel electrophoresis . DNA fragments carrying gene sequences coding for the variable or constant region of k chains were detected by hybridization with purified, 125I-labeled, whole MOPC 321 K MRNA and with its 3'-end half . The pattern of hybridization was completely different in the genomes of embryo cells and of the plasmacytoma . The pattern of embryo DNA showed two components, one of which (molecular weight=6.0 million) hybridized with C-gene sequences and the other (molecular weight=3.9 million) with V-gene sequences . The pattern of the tumor DNA showed a single component that hybridized with both V-gene and C-gene sequences and that is smaller (molecular weight=2.4 million) than either of the components in embryo DNA . The results were interpreted to mean that the Vk and Ck genes, which are some distance away from each other in the embryo cells, are joined to form a contiguous polynucleotide stretch during differentiation of lymphocytes . Such joining occurs in both of the homologous chromosomes . Relevance of these findings with respect to models for V-C gene joining, activation of a specific V k gene, and allelic exclusion in immunoglobulin gene loci is discussed.

J Clin Pathol, 1976 Oct, 29(10), 941 - 8
Studies on the production of enterotoxins by Bacillus cereus; Turnbull PC; Evidence is presented for the existence of three distinct enterotoxins detected in concentrated cell-free culture filtrates of selected Bacillus cereus strains . The first was a product capable of stimulating the adenylate cyclase-cyclic-AMP system in intestinal epithelial cells and, possibly through this, causing fluid accumulation in ligated ileal sections ("loops") of young rabbits . This was elaborated by a strain isolated from an incident of diarrhoea and which caused diarrhoea in 6 of 10 monkey feedings . The second was tentatively identified as a factor which caused fluid accumulation in rabbit loops but not, apparently, through stimulation of the adenylate cyclase-cyclic-AMP system; this was elaborated by a strain isolated from raw rice which failed to produce symptoms in eight monkey feedings . Together, the behaviour of these two factors indicates that diarrhoea caused by B . cereus enterotoxin may be a cyclic-AMP-mediated event . The third, here referred to as "pyogenic toxin", caused severe tissue damage in the ileal mucosa and was elaborated by a strain isolated from a brain abscess . A factor produced by a strain isolated from an outbreak of vomiting which caused vomiting in 10 of 24 monkey feedings could bot be detected in tests reported here but appears to be a fourth enterotoxin type . Cytopathic effects in tissue cultures, suckling mouse tests, and assays of glycerol production by fat cells were not found to be of value in the detection of any of the enterotoxins.

J Clin Pathol, 1976 Oct, 29(10), 938 - 40
Identification of a novel enterotoxigenic activity associated with Bacillus cereus; Melling J et al.; A strain of Bacillus cereus isolated from a food poisoning outbreak characterized by vomiting has been shown to be capable of causing vomiting when cultures grown on rice, but not other media, were fed to Rhesus monkeys . In contrast, a strain isolated from a diarrhoeal outbreak produced diarrhoea, but not vomiting, when grown on various media in similar feeding trials . Furthermore, culture filtrates from the diarrhoeal strain caused fluid accumulation in ligated rabbit ileal loops whereas those from the vomiting strain did not . It is proposed that at least two enterotoxins are involved, one responsible for the vomiting and one for the diarrhoeal symptoms.

J Med Chem, 1976 Oct, 19(10), 1228 - 31
Structure of the peptide antibiotic polypeptin; Sogn JA; Polypeptin, a basic peptide antibiotic isolated from Bacillus circulans, was separated into two components by countercurrent distribution . The two components, polypeptin A and polypeptin B, had identical amino acid compositions but varied in the structure of the hydroxy acid constituent attached to the alpha-amino group of the peptide chain . Polypeptin A contained 3-hydrosy-4-methylhexanoic acid and polypeptin B contained 3-hydrosy-5-methylhexanoic acid . T-HE STEROCHEMISTRY OF THESE HYDROXY ACIDS WAS NOT DETERMINED . Studies involving partial acid hydrolysis and chemical synthesis are consistent with the lactone sturcture for polypeptin A . Polypeptin B differs only in the position of the methyl group in the hydroxyacyl moiety.

Can J Biochem, 1976 Oct, 54(10), 854 - 65
Evidence against the involvement of adenosine 3',5'-cyclic monophosphate in glucose inhibition of beta-galactosidase induction in Bacillus megaterium; Yeung KH et al.; When Bacillus megaterium cells are grown on D-galactose as the sole carbon source, the cells actively synthesize beta-galactosidase (beta-D-galactoside galactohydrolase, EC 3.2.1.23) . However, D-galactose, when added to a glucose-grown culture, did not induce beta-galactosidase, apparently because of the glucose inhibition of the transport of galactose . On the other hand, when glucose was added to a galactose-grown culture, the transport of galactose continued at a reduced but significate rate, whereas further synthesis of beta-galactosidase was halted . Adenosine 3',5'-cyclic monophosphate (camp) or guanosine 3',5'-cyclic monophosphate (Cgmp) did not relieve the glucose inhibition of beta-galactosidase synthesis in the preinduced culture . A method which gave a reproducible assay of c{32P}AMP in Escherichia coli did not detect cAMP or cGMP in a B . megaterium culture undergoing beta-galactosidase induction, but revealed the extracellular accumulation of two unknown phosphorylated compounds . Cell-free extracts prepared from galactose-grown cells did not catalyze the degradation of cAMP or cGMP.

J Gen Microbiol, 1976 Oct, 96(2), 415 - 22
Production of gramicidin S synthetases by Bacillus brevis in continuous culture; Matteo CC et al.; The effects of different nutrient limitations on the production of the two enzymes of gramicidin S biosynthesis were studied during continuous culture of Bacillus brevis . Gramicidin S synthetases I and II were produced in the chemostat under carbon, nitrogen, phosphorus or sulphur limitation . The growth rate, rather than the nature of the limitation, was the major controlling factor in regulating the level of the gramicidin S synthetases . Synthetase production was low at high dilution rates (0.45 to 0.50 h-1) but increased as the dilution rate was lowered . The highest specific activities occurred at dilution rates that were different for each type of limitation: 0.40 h-1 for nitrogen, 0.32 h-1 for carbon, 0.24 h-1 for sulphur and 0.20 h-1 for phosphorus . Phosphorus limitation gave the highest specific activities . At low dilution rates (0.10 to 0.15 h-1), enzyme activities were again low . Sporulation occurred under carbon limitation, but at a lower dilution rate than that which supported optimal gramicidin S synthetase formation . The specific productivity of the synthetases in the chemostat was higher than the highest productivity obtained in batch growth.

Ann Microbiol (Paris), 1976 Oct, 127B(3), 295 - 307
{Studies on the mechanism of interaction of glutaraldehyde with microorganisms (author's transl)}; Navarro JM et al.; The antimicrobial action of glutaraldehyde on Candida lipolytica increases with reagent concentration, pH and duration of contact . The simultaneous study of the size distribution of the particles shows an agglutination of the cells . This result is confirmed by direct observation of the cells by electron microscopy . Cell agglutination also occurs with other microorganisms (Saccharomyces carlsbergensis, Bacillus megaterium, Escherichia coli) and increases their settling rate . The formation of such intercellular bonds seems to confirm the hypothesis of a preferential action of glutaraldehyde on the outer layers of the cells . The effect of pH on the antimicrobial action of glutaraldehyde is accounted for by the fact that this reagent does not act, under neutral and alcaline conditions, as a dialdehyde but as an unsaturated polymer . This mechanism also accounts for various observations made on microorganisms-glutaraldehyde interactions.

Biochim Biophys Acta, 1976 Sep 27, 441(3), 423 - 32
Selective utilization of endogenous unsaturated phosphatidylcholines and diacylglycerols by cholinephosphotransferase of mouse lung microsomes; Kottgen E et al.; In the presence of CMP, cholinephosphotransferase of mouse lung microsomes catalyzes the conversion of endogenous phosphatidylcholines into 1,2-diacyl-sn-glycerols and CDPcholine . 2 . In this conversion cholinephosphotransferase shows a distinct preference for those molecular species of phosphatidylcholine which contain an unsaturated fatty acid . The enzyme hardly utilizes endogenous depalmitoylglycerophosphocholine as a substrate . 3 . Membrane-bound 1,2-diacyl-sn-glycerols were also prepared by treatment of mouse lung microsomes with a pure phospholipase C from Bacillus cereus . These 1,2-diacyl-sn-glycerols were subsequently utilized as substrate by cholinephosphotransferase in the formation of phosphatidylcholine . In the latter reaction, cholinephosphotransferase exhibited a pronounced preference for unsaturated 1,2-diacyl-sn-glycerols and hardly utilized the endogenous 1,2-depalmitoyl-sn-glycerol . 4 . The low affinity of cholinephosphotransferase for either dipalmitoylglycerophosphocholine or 1,2-dip

Eur J Biochem, 1976 Sep 15, 68(2), 581 - 9
Membrane-bound DD-carboxypeptidase and transpeptidase activities from Bacillus megaterium KM at pH 7 . General properties, substrate specificity and inhibition by beta-lactam antibiotics; Marquet A et al.; 1 . The membranes from Bacillus megaterium KM contained a DD-carboxypeptidase with optimum activity under the following conditions: pH 7; ionic strength, 1.3 M; temperature, 40 degrees C and below 20 degrees C . It did not require any divalent cation, but was inactivated by Cu2+ and Hg2+ . It was stimulated by 2-mercaptoethanol and low concentrations of p-chloromercuribenzoate . 2 . The membrane preparation also catalyzed a simple transpeptidation reaction using as carboxyl acceptors D-alanine or glycine . 3 . The conditions for optimum activity, temperature-inactivation, temperature-dependence of the activity, carboxyl donor specificity, sensitivity to beta-lactam antibiotics, and insensitivity to potential peptide inhibitors of both enzyme activities, was identical . The DD-carboxypeptidase showed inhibition by D-alanine and Ac2-L-Lys-D-Ala . 4 . The inhibition by beta-lactam antibiotic was reversible for both enzymic activities and the time-dependence for their recovery was identical . 5 . The DD-carboxypeptidase was very sensitive to changes in the configuration and size of the side-chains of the C-terminal dipeptide of the substrate . Amino acid residues at the C-terminus that precluded the peptide from being a DD-carboxypeptidase substrate were not acceptors in the transpeptidation reaction . Dipeptides were not acceptors for the 'model transpeptidase' . 6 . It is suggested that both activities are catalysed by the same enzyme molecule, whose physiological role is not the formation of peptide crosslinks during peptidoglycan biosynthesis.

Biochim Biophys Acta, 1976 Sep 14, 445(2), 350 - 63
Lysine- and lysine-plus-threonine-inhibitable aspartokinases in Bacillus brevis; Hitchcock MH et al.; Two aspartokinase (ATP:L-aspartate 4-phosphotrasferase, EC 2.7.2.4) enzyme activities have been identified and partially purified from Bacillus brevis . Aspartokinase I is subject to both inhibition and repression by lysine, and has a molecular weight in the region of 110 000 . Aspartokinase II is a lysine-stabilised enzyme, inhibited multivalently by lysine plus theonine and has a molecular weight in the region of 95 000 . This attern of aspartokinase activity has not been described previously and is unusual in that one end product (lysine) regulates two isoenzymes catalysing the first reaction of a branced biosynthetic pathway . In the absence of lysine, aspartokinase II changes to a more unstable non-inhibitable enzyme . Both enzymes are stabilised by sulphydryl reducing agents and have similar affinities for ATP, aspartate and lysine . However, there is no evidence for a view that they are products of a common gene . Problem concerned with the regulation of aspartokinase activities in Bacillus species are discussed.

C R Acad Sci Hebd Seances Acad Sci D, 1976 Sep 13, 283(4), 371 - 3
{The effect of repeated amputations on planarian regeneration in the presence of the heat-stable toxin from Bacillus thuringiensis}; Ziller C; The exotoxin of Bacillus thuringiensis, which is an inhibitor of RNA synthesis, inhibits Planarian regeneration . Planarians which have been cut twice at the same level are able to regenerate after the second section in the presence of the toxin . This indicates that the first amputation stimulates a synthesis of stable RNAs which thus are available for regeneration at the moment of the second section.

JAMA, 1976 Sep 13, 236(11), 1264 - 6
Gram-negative bacillary meningitis therapy . Polyradiculitis following intralumbar aminoglycoside administration; Hollifield JW et al.; Gram-negative bacillary meningitis is a serious threat to patients with head trauma or altered immune systems, or those who have had neurosurgical procedures . The aminoglycoside antibiotics administered systemically and into the cerebrospinal fluid (CSF) have proved useful in the treatment of these infections . Intralumbar and parenteral gentamicin sulfate and tobramycin sulfate were administered to an acromegalic woman with Klebsiella meningitis . She had objective evidence of adhesive arachnoiditis and symptoms of polyradiculitis . Cessation of the aminoglycoside administration resulted in symptomatic improvement, and reinstitution of therapy caused a recurrence of neurologic symptoms . Polyradiculitis may be a complication of the intralumbar administration of aminoglycosides.

Biochim Biophys Acta, 1976 Sep 7, 443(3), 348 - 59
The relationship between environmental temperature, cell growth and the fluidity and physical state of the membrane lipids in Bacillus stearothermophilus; McElhaney RN et al.; A definite and characteristic relationship exists between growth temperature, fatty acid composition and the fluidity and physical state of the membrane lipids in wild type Bacillus stearothermophilus . As the environmental temperature is increased, the proportion of saturated fatty acids found in the membrane lipids is also markedly increased with a concomitant decrease in the proportion of unsaturated and branched chain fatty acids . The temperature range over which the gel to liquid-crystalline membrane lipid phase transition occurs is thereby shifted such that the upper boundary of this transition always lies near (and usually below) the temperature of growth . This organism thus possesses an effective and sensitive homeoviscous adaptation mechanism which maintains a relatively constant degree of membrane lipid fluidity over a wide range of environmental temperatures . A mutant of B . stearothermophilus which has lost the ability to increase the proportion of relatively high melting fatty acids in the membrane lipids, and thereby increase the phase transition temperature in response to increases in environmental temperature, is also unable to grow at higher temperatures . An effective homeoviscous regulatory mechanism thus appears to extend the growth temperature range of the wild type organism and may be an essential feature of adaptation to temperature extremes . Over most of their growth temperature ranges the membrane lipids of wild type and temperature-sensitive B . stearothermophilus cells exist entirely or nearly entirely in the liquid-crystalline state . Also, the temperature-sensitive mutant is capable of growth at temperatures well above those at which the membrane lipid gel to liquid-crystalline phase transition is completed . Therefore, although other evidence suggests the existence of an upper limit on the degree of membrane fluidity compatible with cell growth, the phase transition is completed . Therefore, although other evidence suggests the existence of an upper limit on the degree of membrane fluidity compatible with cell growth, the phase transition upper boundary itself does not directly determine the maximum growth temperature of this organism . Similarly, the lower boundary does not determine the minimum growth temperature, since cell growth ceases at a temperature at which most of the membrane lipid still exists in a fluid state . These observations do not support the suggestion made in an earlier study, which utilized electron spin resonance spectroscopy to monitor membrane lipid lateral phase separations, that the minimum and maximum growth temperatures of this organism might directly be determined by the solid-fluid membrane lipid phase transition boundaries . Evidence is presented here that the electron spin resonance techniques used previously did not in fact detect the gel to liquid-crystalline phase transition of the bulk membrane lipids, which, however, can be reliably measured by differential thermal analysis.

Mikrobiologiia, 1976 Sep-Oct, 45(5), 884 - 7
{Electron microscopic study of intracellular growth of phage specific for Bacillus thuringiensis var . galleriae}; Bespalova IA et al.; Ultrathin sections of Bacillus thuringiensis var . galleriae infected by a specific phage were studied by electron microscopy . Stages of the phage reproduction within the cell, from adsorption of phage particles to liberation of the phage from the cell, have been revealed . Growth of the phage follows that of DNA-containing phages . The yield of the phage is very high.

J Clin Pathol, 1976 Sep, 29(9), 806 - 11
Pseudotumour of the lung caused by infection with Bacillus sphaericus; Isaacson P et al.; A patient is described who suffered from severe chronic asthma complicated by repeated chest infections . She developed a large gelatinous pseudotumour of the lung which was found to be caused by Bacillus sphaericus.

J Bacteriol, 1976 Sep, 127(3), 1568 - 70
Electron microscope studies of conditional spore cortexless mutants of Bacillus sphaericus; Imae Y et al.; Spore cortex of conditional cortexless mutants of Bacillus sphaericus 9602 was not detectable by electron microscopy unless the medium was supplemented with meso-alpha,epsilon-diaminopimelic acid during sporulation . Other spore structures appeared normal . Spore shape was quite irregular in the absence of meso-alpha,epsilon-diaminopimelic acid.

Vopr Virusol, 1976 Sep-Oct, (5), 597 - 601
{Physicochemical characteristics of DNA and the amino acid composition of Bacillus phages}; Polkhovskii VA; From five Bacillus phages nucleic acids were isolated and identified as double-stranded DNA of the AT-type . Their melting point was 85-86.5 degrees C, molecular weight 23.4-43.7X10(6) daltons . The content of DNA in phage particles was 39.9-44.0%, the content of protein 55.7-56.1% . The examination of the amino acid composition in phages 125/50 and 617 revealed a low level of serine and the lack of methionine, while the remaining amino acid composition of Bacillus phages was close to that of E . coli T2 and T3 phages.

Am Rev Respir Dis, 1976 Sep, 114(3), 549 - 54
The pathogenicity of spheroplasts of Mycobacterium tuberculosis; Ratnam S et al.; Guinea pigs were injected with spheroplasts of Mycobacterium tuberculosis and were sacrificed after various intervals to study the state of the organisms and to evaluate the extent of disease . The spheroplasts persisted in the body of the animals for a considerable length of time . Their presence, however, was not associated with either the development of tuberculin hypersensitivity or pathologic changes . It was found that the spheroplasts were pathogenic only when reverted in the animals to bacterial forms . The reversion of the spheroplasts to bacillary state and the subsequent initiation of infection were dependent on the duration of their stay within the host.

Eur J Biochem, 1976 Sep, 68(1), 169 - 76
Occurrence in Bacillus licheniformis of two species of 5-S RNA with multiple differences in primary structure; Raue HA et al.; Bacillus licheniformis was found to contain two species of 5-S RNA . One of these, the primary structure of which has been published previously {H . A . Raue, T.J . Stoof and R.J . Planta (1975) Eur . J . Biochem . 59, 35--42} accounts for 80--90% of the total cellular amount of 5-S RNA . The other one, comprising 10--20% of the total amount, differs in primary structure from the major species at eight positions . All base changes are either purine leads to purine or pyrimidine leads to pyrimidine substitutions . Half of the changes are located within the 5'-terminal part (15 nucleotides) of the molecule, the other half in the 3'-terminal region (22 nucleotides) . At least one of the base changes is in a region which in B . subtilis has been implicated in processing of precursor 5-S RNA . The data are consistent with the existence of a single 5-S RNA cistron with a primary structure different from that of all other 5-S RNA cistrons in B . licheniformis.

J Bacteriol, 1976 Sep, 127(3), 1324 - 30
Ferric hydroxamate transport without subsequent iron utilization in Bacillus megaterium; Arceneaux JE et al.; Iron transport and utilization were examined in Bacillus megaterium Ard1, a mutant that is resistant to the hydroxymate antibiotic A22765 and whose growth is inhibited by the structurally similar hydroxamate Desferal . Rapid, low-level uptake of Desferal-50Fe was observed; such uptake was temperature and energy independent . Gel filtration chromatography of the cytoplasmic fraction of protoplasts labeled with Desferal-55Fe for 30 to 120 s demonstrated only unchanged esferal-55Fe in the cytoplasm . Although B . megaterium Ard1 showed transport of Desferal-59Fe by a process that resembles facilitated diffusion, this organism was unable to transfer iron from this chelate to cellular macromolecules for metabolic use . High-level transport of the ferric hydroxamate schizokinen-59Fe by B . megaterium Ard1 was both temperature and energy dependent . Within 30 s, protoplasts labeled with schizokinen-55Fe contained iron associated with certain macromolecules and in an apparent "pool" of schizokinen-55Fe in the cytoplasmic fraction . Prior transport of Dseferal-55Fe by protoplasts of strain Ard1 did not interfere with subsequent transport and utilization of schizokinen-59Fe . These studies suggest that transport of ferric hydroxamates may occur by a facilitated diffusion-type process; transfer of iron to cellular macromolecules may drive high-level transport of the chelate and may be the step at which energy is required in the iron transport-assimilation process.

Cancer Res, 1976 Sep, 36(9 pt.1), 3255 - 9
Mechanism of Bacillus Calmette-Guérin-induced suppression of metastases in a poorly immunogenic fibrosarcoma; Liotta LA et al.; Bacillus Calmette-Guerin (BCG) reduces the rate of spontaneous pulmonary metastases from a poorly immunogenic fibrosarcoma . To be effective, BCG (1 X 10(6) organisms) must be given in admixture with (1 X 10(6)) tumor cells at the time of transplantation . Reduction of metastasis at this dosage of BCG occurs without a change in the size of the primarytumor or the extent of necrosis within it . Tumors transplanted in admixture with spleen cells from BCG-exposed donors reduced the number of metastases, while spleen cells from normal or tumor-bearing donors had no effect on metastases . Fewer total tumor cells and clumps are collected from the venous effluent of tumors transplanted with BCG than from control tumors . The BCG-treated tumors have more host macrophages intimately associated with the effluent tumor cells then did controls . These data indicate that BCG can inhibit the metastatic potential of a weakly immunogenic fibrosarcoma . The mechanism of this effect appears to be a depression of entry or tumor cells into the tumor vascular channels which may be related to the interaction of tumor cells with BCG-stimulated macrophages.

Can J Microbiol, 1976 Sep, 22(9), 1293 - 9
The respiratory metabolism of Mycobacterium lepraemurium; Kato L et al.; The respiratory metabolism of Mycobacterium lepraemurium isolated from Sprague-Dawley rats lepromata using several substrates was investigated . None of the intermediates of the glycolysis cycle as well as of the tricarboxylic acid cycle except succinate was oxidized by purified whole suspensions of M . lepraemurium . Likewise, many sulfur compounds such as cystine, thiourea, thioacetate, thiodiglycol, mercaptoact and some sulfhydryl compounds, e.g., cysteine, dithioerythritol, dithiorthritol, and penicillamine were readily oxidized by murine bacillary suspensions, whereas thioglycolate, thioglucose, and reduced glutathione were oxidized at a slow rate . Succinate was not or was very poorly oxidized by normal cells probably because of impermeability of the cell wall but the addition of succinate to the cell suspensions frozen for 1 min at -40 degrees C considerably enhanced oxygen uptake over the endogenous value . The oxidation of succinate was unaffected by inhibitors rotenone, atabrine, and amytal but was markedly inhibited by thenoyltrifluoroacetone, antimycin A, 2-N-heptyl-4-hydroxyquinoline-N-oxide, and cyanide . The thiol-binding agents, p-hydroxymercuribenzoate and N-ethylmaleimide were also effective inhibitors of succinate oxidation but the process was not affected by uncouplers dinitrophenol, dibromophenol, pentachlorophenol, and carbonyl-cyanide-m-chlorophenylhydrazone . The results indicated that succinate oxidation by M . lepraemurium was mediated by oxidative enzymes involving an electron transport chain with oxygen as the terminal electron acceptor.

J Thorac Cardiovasc Surg, 1976 Sep, 72(3), 333 - 8
Regional immunotherapy with intrapleural BCG for lung cancer; McKneally MF et al.; A living vaccine, the bacille Calmette Guerin (BCG) strain of Mycobacterium tuberculosis bovis, has been administered in a single postoperative intrapleural dose as a controlled equivalent of postoperative empyema in 38 patients following pulmonary resection for lung cancer . This form of regional immunotherapy is reasonably well tolerated if the vaccine is given in a limited dose and if a follow-up course of isoniazid (INH) is administered . In this report, the technical details of this new therapeutic program are reviewed . The preliminary findings in a randomized prospective clinical train of the technique indicate that patients with Stage I lung cancer are significantly benefitted by the treatment . Patients with more advanced disease are unimproved by this form of therapy.

J Bacteriol, 1976 Sep, 127(3), 1472 - 81
Electron microscope study of sporulation and parasporal crystal formation in Bacillus thuringiensis; Bechtel DB et al.; A comprehensive ultrastructural analysis of sporulation and parasporal crystal development is described for Bacillus thuringiensis . The insecticidal crystal of B . thuringiensis is initiated at the start of engulfment and is nearly complete by the time the exosporium forms . The crystal and a heretofore unobserved ovoid inclusion develop without any clear association with the forespore septum, exosporium, or mesosomes . These observations contradict previous hypotheses that the crystal is synthesized on the forespore membrane, exosporium, or mesosomes . Formation of forespore septa involves densely staining, double-membrane-bound, vesicular mesosomes that have a bridged appearance . Forespore engulfment is subpolar and also involves mesosomes . Upon completion of engulfment and the following cytoplasmic changes occur: decrease in electron density of the incipient forespore membrane; loss of bridged appearance of incipient forespore membrane; change in stainability of incipient forespore, forespore, and mother cell cytoplasms; and alteration in staining quality of plasma membrane . These changes are involved in the conversion of the incipient forespore into a forespore and reflect "commitment" to sporulation.

Appl Environ Microbiol, 1976 Sep, 32(3), 400 - 4
Defined conditions for synthesis of Bacillus cereus enterotoxin by fermenter-grown cultures; Glatz BA et al.; A strain of Bacillus cereus produced high levels of enterotoxin when grown in a semidefined medium in a laboratory scale fermenter . The optimum conditions for enterotoxin synthesis by cultures grown in this medium, which contained Casamino Acids and yeast extract, were found to be: inoculation of vigorously gorwing culture at the 1% level, addition of glucose at a concentration of 1%, control of culture pH at 8.0, incubation at 32 degrees C, use of a moderate stirring rate, and addition of air at low flow rates to minimize foaming . The enterotoxin yield in fermenter-grown cultures was approximately 20 to 50 times higher than the yield obtained in shake flask cultures.

Biochim Biophys Acta, 1976 Aug 18, 442(2), 184 - 96
The restriction endonucleases in Bacillus amyloliquefaciens N strain . Substrate specificities; Shibata T et al.; Two species of restriction endonuclease were isolated by gel filtration and DEAE-cellulose chromatography from a cell-free extract of Bacillus amyloliquefaciens (B . subtilits) N strain; a lower molecular weight endonuclease (endonuclease R.BamNI) and a higher molecular-weight one (endonuclease R.BamNx) . Both of them required only Mg2+ for their activities . Endonuclease R.BamNx introduced a larger number of site-specific scissions in Excherchia coli phage lambda DNA that endonuclease R.BamNI did . Endonuclease R.BamNx cleaved Bacillus phage phi 105C DNA at the specific sites which are classified into two groups: one type of sites is modified by B . amyloliquefaciens H strain in vivo while the other is not affected . It was also active on DNA'S OF E . coli phage T7, lambdadvl, Simian virus 40 (SV40) and colicinogenic factor ColEI and was inactive on DNAs of Bacillus phages phi 29 and M2 . Endonuclease R.BamHI isolated from H strain by Wilson and Young . This endonuclease was active on DNAs of phage lambda, lambdadvl and SV40, adn was inactive on DNAs of phages phi 105C, phi 29, M2 and T7, and ColEI DNA.

J Am Vet Med Assoc, 1976 Aug 15, 169(4), 419 - 20
Naturally occurring Tyzzer's disease as a complication of distemper and mycotic pneumonia in a dog; Poonacha KB et al.; Tyzzer's disease, distemper, and mycotic pneumonia were diagnosed at necropsy of a 6-week-old mixed breed pup . The illness was characterized by lethargy, poor appetite, dysentery, and convulsions, the latter developing just prior to death . The necropsy findings included pale, dilated large intestine and swollen, dark liver . Histologically, multifocal hepatic necrosis, similar to that of Tyzzer's disease in other species of animals, was observed . Bacillus piliformis was found in hepatocytes and necrotic areas of the liver.

Mol Gen Genet, 1976 Aug 10, 147(1), 1 - 12
Conditional mutations in the translational apparatus of Bacillus subtils; Dubnau E et al.; Four temperature sensitive mutants of B . subtilis were isolated by localized mutagenesis in the major ribosomal gene cluster, and charcterized genetically and biochemically . Three are mutations which cause temperature sensitivity in the elongation factor Ef-G, and one which has a similar effect on the elongation factor Ef-Tu . They map in a cluster near strA, with the temperature sensitive mutations in Ef-G mapping between the strA gene and the temperature sensitive mutation in Ef-TU.

Jpn J Microbiol, 1976 Aug, 20(4), 303 - 8
Effects of BCG (Bacillus Calmette-Guérin) vaccines on immune responses in mice . I . Possible effect of BCG on helper T cells; Kitamura Y et al.; The effects of killed and living BCG on antibody production against hamster erythrocytes (HRBC) and the 2, 4, 6-trinitrophenyl (TNP) group were studied in SL mice . Killed and living BCG, each in doses of 0.008 mg, 0.08 mg, 0.8 mg and 8 mg per mouse, were intravenously inoculated 7 days prior to primary immunization with HRBC . Secondary immunization was carried out 28 days later with TNP-HRBC . Anti-HRBC and anti-TNP antibodies were estimated by a hemagglutination test . The results showed that pretreatment with killed or living BCG enhanced the antibody production against both HRBC and TNP . Comparing the effects of these two BCG preparations, it was noted that killed BCG augmented the anti-HRBC antibody production more effectively than living BCG . In regard to the anti-TNP antibody production, living BCG exhibited a greater augmenting effect than killed BCG . This difference in the modes of action of killed and living BCG was remarkable when two groups given 8 mg of killed and living BCG were compared . In addition, it was shown that living BCG at a dose as high as 8 mg was able to augment the anti-TNP antibody production, even in the absence of preceding immunization with HRBC.

J Antibiot (Tokyo), 1976 Aug, 29(8), 813 - 7
Isolation of a new peptide antibiotic complex, B-43 (studies on antibiotics from the genus Bacillus . XV); Shoji J et al.; A new peptide antibiotic complex B-43, active against Gram-positive and Gram-negative bacteria, was isolated from a strain of Bacillus circulans . This antibiotic contains aspartic acid, valine, isoleucine, leucine, phenylalanine and 2,4-diaminobutyric acid . It seems to be related to polypeptin and antibiotic complex 4205, but differs in that it contains aspartic acid residue.

J Antibiot (Tokyo), 1976 Aug, 29(8), 809 - 12
Isolation of a new peptide antibiotic, 339-29 (studies on antibiotics from the genus Bacillus . XIV); Shoji J et al.; A new antibiotic named 339-29, active against Gram-positive bacteria was isolated from a strain of Bacillys pumilus . The hydrochloride is soluble in water and aqueous alcohols . The antibiotic is a basic peptide containing valine (3), isoleucine (1), Leucine (2), tyrosine (1), lysine (3) and a fatty acid.

Appl Environ Microbiol, 1976 Aug, 32(2), 312 - 4
Heat-induced requirements for sucrose or magnesium for expression of heat resistance in Bacillus cereus forespores; Busta FF et al.; The addition of 0.6 M sucrose of 0.016 M Mg2+ to the enumeration medium was required for early expression of heat resistance (10 min at 70 degrees C) in stage V Bacillus cereus forspores . The addition of Mg2+ to the sporulation medium did not remove this requirment for sucrose of Mg2+ . The heat damage did not affect forespore germination or outgrowth, but injured cells in the absence of sucrose or Mg2+ were not capable of cell division . The heat-induced sublethal damage apparently affected the forspore component(s) that could be repaired or was capable of normal function in the presence of added Mg2+ or sucrose.

J Gen Microbiol, 1976 Aug, 96(2), 381 - 90
Commitment to sporulation in Bacillus megaterium and uptake of specific compounds; Cooney PH et al.; Commitment of Bacillus megaterium cells to continue the sporulation process was tested at different times during the developmental period with respect to either addition of different carbon sources (sugars or amino acids) or dilution into media containing these . Organisms grown in minimal medium containing sucrose as sole carbon source were committed earliest with respect to aspartic or glutamic acid as sole carbon source, later with respect to fructose, glucose, glycerol or sucrose, and latest with respect to nutrient medium supplemented with casein hydrolysate . Addition of both aspartate and a carbohydrate resulted in later commitment than addition of either compound alone . The initial uptake rates of aspartate, glutamate, glucose and sucrose increased toward the end of growth in complex medium (but not in minimal medium for glucose and sucrose) and then decreased during the developmental period.

J Gen Microbiol, 1976 Aug, 96(2), 268 - 76
Inhibition of RNA synthesis in Chlorella pyrenoidosa and Bacillus megaterium by the pine-blight toxin, dothistromin; Harvey AM et al.; Dosthistromin, an anthraquinone derivative produced by the pine-blight fungus, Dothistroma pini, inhibits the growth of Chlorella pyrenoidosa and Bacillus megaterium . At growth inhibitory concentrations, dothistromin strongly inhibits incorporation of {3H}uridine into RNA of both species . With B . megaterium, marked inhibition of {3H}uridine incorporation is apparent within 5 min of addition of dothistromin, but only a slight inhibition of {3H}thymidine incorporation into the DNA-containing fraction or of {14C}leucine incorporation into protein is detectable after 10 min.

J Bacteriol, 1976 Aug, 127(2), 829 - 31
Low-frequency, pbsi-mediated plasmid transduction in Bacillus pumilus; Bramucci MG et al.; Three bacteriophages were tested for ability to transduce the plasmid of pPL10 between W mutant derivatives of Bacillus pumilus NRS 576 . Phage PBP1- and PMB1-generated plasmid transductants occurred at about 10% the frequency of transductants for a chromosome marker . Phage PBS1-generated plasmid transductants occurred at less than 0.1% the frequency of transductants for a chromosome marker . Possible reasons for the extremely reduced capacity of PBS1 to generate plasmid transductants are discussed.

J Urol, 1976 Aug, 116(2), 180 - 3
Intracavitary Bacillus Calmette-Guerin in the treatment of superficial bladder tumors; Morales A et al.; Patients with recurrent superficial bladder tumors have been treated by vesical and intradermal administration of Bacillus Calmette-Guerin . The pattern of recurrence in 9 patients has been altered favorably . Although the findings are still preliminary they appear to hold promise of a new therapeutic approach to the treatment of a group of neoplasms for which effective therapy is still lacking.

South Med J, 1976 Aug, 69(8), 986 - 93
The histopathologic diagnosis of leprosy; Mansfield RE et al.; The objective of this paper is to encourage histopathologists to recognize and diagnose leprosy so that it can be treated effectively at the earliest stage possible, before irreversible deformities result . Recognition of the earliest form of leprosy-indeterminate-is emphasized . Histopathologic descriptions are made and illustrated for the principal types of leprosy . Emphasis is placed on (1) the need for dependable acid-fast staining of leprosy bacillus, (2) the non-specific infiltrate in indeterminate leprosy, and (3) the involvement of nerves in all types of leprosy.

Appl Environ Microbiol, 1976 Aug, 32(2), 217 - 21
Effects of added germination agents on loss of optical density in electron-irradiated spores; Stogaitis G et al.; Spores of Bacillus megaterium ATCC 14581, subjected to partial-cell iradiation, were exposed to either lysozyme, H2O2, or glucose in an attempt to reduce or eliminate the nonmonotonic behavior in curves of percentage of germination versus energy, obtained when such spores were resuspended in phosphate buffer alone . Except at the lower doses . H2O2 effectively eliminated this anomalous dip in these curves, whereas lysozyme amplified it greatly . Glucose was generally ineffective . Coinciding with the increases in optical density when lysozyme was present was the formation of an occluding product.

Arch Microbiol, 1976 Aug, 109(1-2), 105 - 8
Effect of the pH of culture medium on the alkalophilicity of a species of Bacillus; Koyama N et al.; The amino acid incorporation and alpha-amino-isobutyric acid (AIB) uptake of an alkalophilic Bacillus grown at pH 8.2 (the pH 8-bacteria) were much less pH dependent (less alkalophilic) than those of the organisms grown at pH 10.0 (the pH 10-bacteria), respectively . The rate of AIB uptake of the pH 10-bacteria was almost the same as that of the pH 8-bacteria, while the rate of amino acid incorporation of the pH 10-bacteria was higher than that of the pH 8-bacteria in alkaline environments . The colloidal titration with clupein showed that the amount of negative charge on the pH 10-bacteria was greater than that of the pH 8-bacteria in alkaline environments . Considerable difference in protein composition was observed between the membranes of the pH 8- and 10-bacteria while no difference was observed in phospholipid composition.

Biochemistry, 1976 Jul 27, 15(15), 3342 - 6
Enzyme hyperspecificity . Rejection of threonine by the valyl-tRNA synthetase by misacylation and hydrolytic editing; Fersht AR et al.; Valyl-tRNA synthetase from Bacillus stearothermophilus activates thereonine and forms a 1:1 complex with threonyl adenylate, but it does not catalyze the net formation of threonyl-tRNAVal at pH 7.78 and 25 degrees C in the quenched flow apparatus it decomposes at a rate constant of 36s-1 . During this process there is a transient formation of Thr-tRNAVal reaching a maximum at 25 ms and rapidly falling to zero after 150 ms . At the peak, 22% of the (14C) threonine from the complex is present as (14C) Thr-tRNA . The reaction may be quenched with phenol and the partially mischarged tRNA isolated . The enzyme catalyzes its hydrolysis with a rate constant of 40s-1 . The data fit a kinetic scheme in which 62% of the threonine from the threonyl adenylate is transferred to the tRNA . This may be compared with the rate constant of 12s-1 at which 84% of the valine is transferred to tRNAVal from the enzyme-bound valyl adenylate, and the rate constant of 0.015s-1 for the subsequent hydrolysis of Val-tRNAVal . Inhibition studies indicate a distinct second site for hydrolysis . The translocation of the aminoacyl moiety between the two sites could be mediated by a transfer between the 2'-and 3'-OH groups of the terminal adenosine fo the tRNA . The hyperspecificity of the enzyme is based on discriminating between the two competing substrates twice: once against the undesired substrate in the synthetic step, and once against the desired substrate in the destructive step.

Arch Otorhinolaryngol, 1976 Jul 20, 212(3), 203 - 11
A clinical study on the ototoxic effects of tobramycin; Lehmann W et al.; In a clinical, prospective, non-randomized study, tobramycin has shown its high antibacterial activity in Gram-negative bacillary infections . The incidence of ototoxic side effects was very low: none of 18 patients studied before, at the end and 9 months after completion of tobramycin therapy showed any change in pure tone audiograms . Five of these 18 patients had renal impairment: 3 of them developed subjective and objective evidence of vestibular dysfunction, as demonstrated by caloric vestibulometry . However, these bilateral vestibular lesions all proved to be reversible a few weeks after cessation of therapy . Despite the low incidence and the reversibility of the ototoxic side effects, it seems advisable to monitor patients on long-duration tobramycin therapy for signs of vestibular dysfunction, particularly in cases of renal insufficiency.

J Biol Chem, 1976 Jul 10, 251(13), 4102 - 10
The hydrophobic membrane penicillinase of Bacillus licheniformis 749/C . Characterization of the hydrophilic enzyme and phospholipopeptide produced by trypsin cleavage; Yamamoto S et al.; The membrane penicillinase of Bacillus licheniformis 749/C is a phospholipoprotein carrying extra residues of asparagine or aspartate, serine, glutamine or glutamate and glycine not present in the exoenzyme (Yamamoto, S., and Lampen, J.O . (1976) J . Biol . Chem . 251, 4095-4101) . Cleavage of the membrane enzyme with trypsin yielded a phospholipipopeptide and a hydrophilic penicillinase differing from exopenicillinase only by the absence of the NH2-terminal lysine residue . Phosphatidylserine was isolated from a pronase digest of the phospholipopeptide . The partial sequence of the phospholipopeptide is: phosphatidylserine-(Ser3, Glx5, Asx7, Gly5)-Asp-Gin-Ser-Lys-COOH with the lysine being the NH2-terminal residue of the usual exoenzyme . The fatty acids present in the membrane enzyme and in the phospholipopeptide had essentially the same composition (predominantly n-16:0, ante iso-17:0, n-18:0, and n-18:1) . These acids were also found in the total membrane lipids, although in very different proportions; thus, the phosphatidic acid residue of the phosphatidylserine is probably formed by the usual synthetic pathway for membrane phospholipids, but some special feature of the process affects the nature of the component fatty acids.

J Biol Chem, 1976 Jul 10, 251(13), 4095 - 101
Purification of plasma membrane penicillinase from Bacillus licheniformis 749/C and comparison with exoenzyme; Yamamoto S et al.; The membrane penicillinase of Bacillus licheniformis 749/C has been demonstrated to be a phospholipoprotein . The homogeneous enzyme gives a positive reaction for phosphorous and for unsaturated fatty acids, has a molecular weight of 33,000 in contrast to 29,000 for the exoenzyme, and contains 8 to 9 additional residues of aspartate or asparagine, 4 to 5 of serine, 7 of glutamate or glutamine, and 4 to 5 of glycine per mole . The COOH-terminal sequence of both membrane and exoenzymes is -Met-Asn-Gln-Lys-COOH; hence the extra peptide portion present in the membrane enzyme is not attached to the COOH-terminus of the exoenzyme . Procedures which readily detected the lysine residue at the NH2 terminus of the exoenzyme did not yield a positive test with the membrane form . The NH2 terminus of the membrane enzyme may be blocked by or linked to the phospholipid . A procedure for the preparation of membrane penicillinase on a large scale and an improved method for purification of the exoenzyme have been developed.

Jugosl Ginekol Opstet, 1976 Jul-Aug, 16(4), 301 - 7
{Vaginal flora in women from Gevgelija}; Demerdziev K; The author gives data on the vaginal flora of women aged 20--64 in the commune of Gevgelia obtained in the course of an early mass detection of the carcinoma of the uterine cervix in this area . The action covered 70.5% of women from urban areas and 85.3% of women from rural areas . The microbiological analysis of vaginal flora was made in two ways: first, by taking smears and by vital staining and classifying them after Jirovec, Peter, and Malik; another preparation was sent to the cytologist and, after fixing and staining, was interpreted microbiologically . The results of both methods are shown in tabular form, according to women's age and residence . They do not differ much between themselves . The purity of vaginal secretion in women from rural areas is higher than that in women from urban areas . The occurrence of Trichomonas vaginalis is twice as frequent in women from rural than from urban areas . There is a significant difference in the existence of Doderlein's bacillus in the postmenopausal period between women from urban and from rural parts of the commune . In women from urban areas over 60 Doderlein's bacillus was present only in 39.28% and in those from rural areas in as many as 63.07% of this group of women.

Prikl Biokhim Mikrobiol, 1976 Jul-Aug, 12(4), 495 - 500
{Relationship between exoprotease activity, spore and crystal formation and virulence of bacteria of the group Bacillus thuringiensis}; Naidenova-Kuzmanova I; Experiments were carried out to investigate the activity, spore and crystal formation and virulence of Bacillus thuringiensis . Bacteria of this group showed different exoprotease activity . Strains of one serotype differed in the level and time of occurrence of peak enzyme activity . Exoprotease activity of the bacteria cannot be indicative of their toxicity because it did not correlate with their virulence . Exoprotease activity can be used to characterize bacteria during their classification and to determine finer differences between strains within one serotype.

Zentralbl Bakteriol {Orig B}, 1976 Jul, 162(1-2), 138 - 44
{Hydrolysis of methoxysubstituted phenylureas, acylanilides and phenylcarbamates by a microbial aryl acylamidase (author's transl)}; Engelhardt G et al.; The phenylurea herbicide linuron is hydrolyzed by Bacillus sphaericus ATCC 12123 quantitatively forming 3,4-dichloroaniline, CO2, and N,O-dimethylhydroxylamine . The inducible enzyme responsible for this hydrolysis was purified to homogeneity as judged by polyacrylamide gel electrophoresis . Its molecular weight was 75 000 +/- 10% . Studies on its substrate specificity showed that either whole cells as the linuron-induced enzyme hydrolyze a large number of herbicidal and fungicidal acylanilides, the methoxysubstituted phenylureas and the phenylcarbamate propham at the carbonyl-aniline bond . This would classify the enzyme as an aryl acylamidase (E.C . 3.5.1) . Hydrolysis of phenylamides by whole cells and by the enzyme is inhibited by different methylcarbamate and organophosphorus insecticides . Inhibition of hydrolysis of linuron by the aryl acylamidase by methylcarbamates is a competitive one.

Nord Vet Med, 1976 Jul-Aug, 28(7-8), 381 - 4
The detection of penicillin in buttermilk; Pekkanen TJ et al.; When 59 buttermilk samples were tested for the presence of antibiotics or chemostherapeutic substances by a method using the spores of Bacillus stearothermophilus var . chalidolactis all samples gave a positive reaction corresponding to about 0.01 IU penicillin per ml . When the buttermilk samples were diluted 1:1 (v/v) by a phosphate buffer, pH 6.8, only 2 were positive which were confirmed to contain penicillin . The mean pH of the unbuffered buttermilks was found to be 4.53 +/- 0.11 and when buffered 6.46 +/- 0.05 . An in vitro addition of benzylpenicillin to buttermilk, to a concentration of 0.01 IU per ml, resulted in a significantly higher concentration of penicillin in the casein fraction than in the buttermilk or in the whey (Table I).

Mikrobiologiia, 1976 JUL-AUG, 45(4), 738 - 40
{Oxygen consumption by suspensions of Bacillus anthracoides spores}; Bekhtereva MN et al.; The respiration rate of spore suspensions of Bacillus anthracoides 96 was assayed by mass spectrometry employing a hermetically sealed reaction vessel constructed for this purpose . The rate of respiration was found to depend on the method of preparing suspensions, the duration of their storage at +4 degrees C, the physiological state of spores, and the action of a disinfectant containing chlorine on them.

Mikrobiologiia, 1976 JUL-AUG, 45(4), 690 - 4
{Ultrastructure of bacteriophages specific for Bacillus thuringiensis var . galleriae}; Rautenshtein RI et al.; Fine structure of the phage specific for Bacillus thuringiensis var . galleriae was studied . The head of the phage is of the elongated hexagonal shape, 600 X 360 A . The tail is 400 X 40 A large and has a thickening in the basal part (70 X 120 A) . A so-called "collar" structure is found at the site where the head joins the tail . This a disc-shaped plate with a diameter of 120 A and a hole in the centre; 12 fillaments are attached to it radially and terminated with tines . It is possible that the structure is necessary for adsorption of the phage on the cell.

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