J Exp Biol, 1979 Dec, 83, 293 - 304 Mechanism of inhibition of active potassium transport in isolated midgut of Manduca sexta by Bacillus thuringiensis endotoxin; Harvey WR et al.; After incubation at pH 10 or higher, Bacillus thuringiensis spores and endotoxin, at concentrations above 0.1 IU/ml, affected transport parameters in the isolated midgut of Manduca sexta larvae . (Toxic activity was lost during roughly 1 week at pH 11.) About 60% of the short-circuit current was inhibited, and the remainder was reversibly inhibited by anoxia . Electrical resistance was reduced by about 55% and oxygen uptake stimulated by about 30% . Influx of potassium from blood-side to lumen-side ('active' flux) was unaffected but flux in the reverse direction was nearly tripled . These results suggest that hydrolysis of the toxin yields an inhibitor of potassium transport, presumably a polypeptide . It is argued that inhibition is not primarily by uncoupling of oxidative phosphorylation, but instead by interference with an active depression of the efflux of potassium from lumen-side to blood-side. Nucleic Acids Res, 1979 Nov 24, 7(6), 1429 - 44 A thermostable, sequence-specific restriction endonuclease from Bacillus stearothermophilus: BstPI; Pugatsch T et al.; A restriction endonuclease, BstPI, was purified from a strain of B . stearothermophilus, and its cleavage specificity was determined . The enzyme cleaves at palindromic sites of the general structure: (Formula: see text) where N.N' can be any base pair . It produces phosphorylated 5'-termini which are single stranded over a length of 5 nucleotides . Ends generated by cleavage with BstPI can be rejoined by DNA ligase. Mol Biol (Mosk), 1979 Nov-Dec, 13(6), 1230 - 6 {Using a combined transcription-translation system for determining the role of cryptic plasmids of Bacillus thuringiensis}; Debabov VG et al.; The possibility of entomocyde crystal protein synthesis was studied using a heterological cell-free system with Bacillus thuringiensis plasmid DNA as template . The high level of template activity is usual for Bac . thuringiensis plasmid DNA . Immunochemical studies of the in vitro synthesized polypeptides showed that Bac . thuringiensis plasmid DNA does not direct crystal protein synthesis. Can J Microbiol, 1979 Nov, 25(11), 1227 - 31 Comparative studies of the mosquito-larval toxin of Bacillus sphaericus SSII-1 and 1593; Myers P et al.; Two strains of Bacillus sphaericus . SSII-1 and 1593, were bioassayed for toxic activity against second-instar larvae of the mosquito Culex pipiens quinquefasciatus . It was found that strain 1593 developed a level of toxicity 3000 times that of strain SSII-1 . Although the toxic activity of B . sphaericus SSII-1 was relatively unchanged throughout growth, an increase in activity of strain 1593 occurred as the bacteria began to sporulate . Strain differences were examined by (i) growth cycle experiments, (ii) bioassays of the toxicity of oligosporogenous mutants, and (iii) manganese limitation experiments . The toxin of strain 1593 was shown to be more stable than that of strain SSII-1 . Unlike the spores of strain SSII-1, the spores of B . sphaericus 1593 were found to be highly toxic . Thin sections of SSII-1 or 1593 cells did not reveal the presence of any inclusion body that might be related to toxicity. J Assoc Off Anal Chem, 1979 Nov, 62(6), 1247 - 50 Microbiological determination of penicillin G, ampicillin, and cloxacillin residues in milk; Vilim AB et al.; A fast cylinder plate microbiological method was developed for the quantitative determination of penicillin G, ampicillin, and cloxacillin in milk . Agar plates seeded with stable spores of Bacillus stearothermophilus var . calidolactis were used and incubated at 64 degrees C for 4 1/2 hr . Standard curves were obtained for the following ranges of concentration of antibiotics: 0.004-0.064 IU penicillin G/mL, 0.0025-0.04 microgram ampicillin/mL, and 0.03-0.48 microgram cloxacillin/mL . The method is suitable for detecting penicillin residues in milk and for quantitative milk-out studies of the above antibiotics used in treatment of bovine mastitis. Am J Clin Pathol, 1979 Nov, 72(5), 882 - 4 Septicemia caused by Pseudomonas paucimobilis; Slotnick IJ et al.; A case of septicemia caused by Pseudomonas paucimobilis is reported . This represents the first definitive case of disease produced by this yellow-pigmented, nonfermentative bacillus. J Bacteriol, 1979 Nov, 140(2), 699 - 706 Converting bacteriophage for sporulation and crystal formation in Bacillus thuringiensis; Perlak FJ et al.; Bacteriophage TP-13, a converting phage for sporulation and crystal formation in Bacillus thuringiensis, was isolated from soil . The phage converted anoligosporogenic (sporulation frequency, 10(-8), acrystalliferous mutant to spore positive, crystal positive at a high frequency . Each plaque formed by TP-13 in a lawn of sensitive cells contained spores and crystals . These spores were heat stable, and each one was capable of producing a plaque from which TP-13 could be reisolated . Conversion of cells to sporulation and crystal formation was independent of the ho-t used for TP-13 propagation . When converted cells were cured of TP-13, they lost the ability to produce spores and crystals . Incubation of TP-13 with antiserum prepared against purified phage particles prevented conversion . TP-13 has some characteristics similar to those of SP-15 and PBS-1, including large size, morphology, and adsorption specificity of motile cells . TP-13 mediated generalized transduction in several strains of B . thuringiensis at frequencies of 10(-6) to 10(-5) . Comparison of cotransduction values indicated that TP-13 transduced considerably larger segments of deoxyribonucleic acid than CP-51 or TP-10, two other transducing phages for B . thuringiensis. Cancer Treat Rep, 1979 Nov-Dec, 63(11-12), 2039 - 41 Phase I study of iv administration of methanol extraction residue of Bacillus Calmette-Guérin; Vogl SE et al.; Four weekly iv doses, ranging from 10 to 640 microgram/m2, of the methanol extraction residue of bacillus Calmette-Guerin (MER) were given to 22 patients with advanced cancer in an effort to determine a maximum tolerated dose . Fever and chills for 24-48 hours were common at doses greater than 80 microgram/m2 but were prolonged for 3 days to 9 weeks at 640 microgram/m2 . While moderate decreases in pulmonary vital capacity were seen at all dose levels, diffusing capacity decreased consistently only at higher doses . However, no roentgenographic changes suggestive of multiple lung granulomas were observed, such as were produced in dogs and mice by iv administration of MER . Autopsy failed to reveal granulomas in three patients . There were no objective tumor responses, nor were consistent changes observed in blood counts or skin tests to recall antigens . MER can be given iv with acceptable acute toxicity in doses up to 640 microgram/m2, at which point fever may be unduly prolonged . Patients given MER iv must be followed carefully for the development of interstitial pneumonitis. Br J Cancer, 1979 Nov, 40(5), 736 - 42 Immunotherapy using BCG during remission induction and as the sole form of maintenance in acute myeloid leukaemia; Summerfield GP et al.; Thirty-two adults with acute myeloid leukaemia (AML) were randomized to receive, from the time of diagnosis, either chemotherapy alone (C group) or chemotherapy plus Bacille Calmette-Guerin vaccine (BCG) (C+I group) . After remission induction and consolidation, chemotherapy was stopped in both groups but BCG was continued in the C+I group . The overall survival of the C+I group was significantly increased (P less than 0.05) . There was no significant increase in the duration of first remission in the C+I group (0.05 less than P less than 0.1) nor in the time from first relapse to death (0.05 less than P less than 0.1) . There was no significant difference in the incidence of first or second remissions, and the time taken to enter remission did not differ significantly between the two groups . Comparison with the results of other trials suggests that the use of maintenance chemotherapy in addition to immunotherapy produces longer remissions . Five patients in the C group developed leukaemic central-nervous-system (CSN) involvement, in comparison with none in the C+I group . CNS relapse did not produce a significant decrease in remission length (P greater than 0.1) but reduction in survival after CNS relapse was highly significant (P = 0.001) . These results suggest that administration of BCG from an early stage in the treatment of AML may protect the CNS against leukaemic infiltration and therefore serve as a simple, innocuous form of CNS prophylaxis. J Maxillofac Surg, 1979 Nov, 7(4), 293 - 8 Cancrum oris; Smith I; Cancrum oris is a gangrenous infection of the oral cavity . Debilitating diseases predispose to the condition . The exact bacteriology is uncertain although Vincent's Spirochaete and the fusiform bacillus in symbiosis have been considered to be the actual cause of the condition . The presentation of the disease is variable, for the condition may affect the soft tissues of the face or the bones of the facial skeleton or both concurrently . This disease has a high mortality rate . If recovery from toxaemia ensues, subsequent healing becomes a lengthy affair because of the destructive nature of the condition . Since the introduction of antibiotics as a form of therapy, a higher survival rate has been achieved . Because of the nature of the condition the morbidity rate is exceptionally high with functional disturbances and disfigurement being a legacy of the disease . Reconstructive procedures are generally required at some subsequent stage. J Biochem (Tokyo), 1979 Nov, 86(5), 1169 - 77 Studies on the allosteric nature of acetate kinase from Bacillus stearothermophilus; Nakajima H et al.; Fructose 1,6-bisphosphate (FBP) stimulates the reaction of Bacillus stearothermophilus acetate kinase (AK) . FBP changes the reaction curve for ATP from a sigmoidal type to a Michaelis-Menten one . The binding of FBP to AK was studied by an equilibrium dialysis method and by measuring changes in fluorescence . The extent of binding of FBP to the enzyme paralleled its activation . In addition, the binding constant for FBP increased in the presence of substrate, ATP . These results suggest that FBP is an allosteric activator of B . stearothermophilus AK . Only two moles of FBP bound to this tetrameric enzyme . No cooperativity was found for the binding of FBP . These observations support the previous conclusion, that a set of two subunits in the tetramer is a unit of the enzymatic function . A model is presented to interpret the sigmoidal kinetics for ATP, the absence of cooperativity for FBP binding, and the allosteric activation by FBP of this enzyme . The kinetic properties of the enzyme can be explained quantitatively by this model. Arch Fr Pediatr, 1979 Nov, 36(9), 926 - 9 {Septic arthritis due to a nontoxigenic diphtheria bacillus}; Guran P et al.; Septic arthritis of the hip in a 2 year old child is described . A nontoxigenic diphtheria bacillus was isolated in large numbers from the articular fluid . The same organism was isolated from excoriated skin lesions of the toes . The bacteriology, epidemiology and pathology of the infection are discussed. Clin Orthop, 1979 Nov-Dec, (145), 237 - 8 Bacillus cereus septic arthritis following arthrography; Robinson SC; Organisms of Bacillus species usually are common laboratory contaminants and nonpathogenic in humans . Recently, however, it has been suggested that cultures growing common Bacillus species may indeed represent significant infections and should not always be disregarded . A 24-year-old man developed Bacillus cereus septic arthritis following routine arthrography, which is an example of a serious orthopedic infection that can be caused by this supposedly nonpathogenic organism. Mikrobiologiia, 1979 Nov-Dec, 48(6), 1093 - 101 {Blooming and destruction of cyanobacteria in the drainage bassin of the hydrogen sulfide spring of Staraya Matsesty}; Gusev MV et al.; The growth of cyanobacteria belonging to the genera Oscillatoria and Anabaena (up to 2.1 x 10(7) filaments per 1 g of wet sample) was found in a water reservoir with a high content of sulfides (up to 9 mM) in Staraya Matsesta throughout the year . The spots of Oscillatoria are located in the spring in more illuminated areas as compared to Anabaena . In the spring, not only spots of actively growing cells were detected, but also accumulations of Oscillatoria cells being destroyed (blue spots) . Water-bloom spots in which Oscillatoria prevailed can transform into the spots of Anabaena . The main accompanying forms in the spots of Anabaena are long thin filaments of the flexibacterial type while short rods are found in the spots where Oscillatoria predominates . Heterotrophic enteric bacteria (48 x 10(4) cells per 1 g), Bacillus, Pseudomonas, and coryneform bacteria were also detected . Green bacteria (Chlorobium) and nonsulfur purple bacteria (Rhodomicrobium) were present in small quantities (16 x 10(3) cells per 1 g) as well as sulfate-reducing bacteria (5--15 x 10(2) cells per 1 g) and thiobacilli (40--60 cells per 1 g) . In the spring, stones were covered with pink spots of spherical motile purple bacteria and with yellow-green spots of filamentous green bacteria . The cyanobacteria from the spots are capable of oxygenated photosynthesis . Fixation of CO2 by them in situ is 0.08 mcg per 1 g of dry sample per hour or 0.06 mcg per 10(6) cells per hour, and is inhibited by 10(-5) M DCMU by 70%. J Gen Microbiol, 1979 Nov, 115(1), 1 - 11 Effects of lysozyme on Bacillus cereus 569: rupture of chains of bacteria and enhancement of sensitivity to autolysins; Westmacott D et al.; Bacillus cereus 569 is known to be resistant to lysis by lysozyme because of the presence of deacetylated glucosamine residues in its peptidoglycan, and cultures continued to grow even in the presence of lysozyme at 200 microgram ml-1 . However, lysozyme caused rupture of the chains of bacteria and promoted the rate of autolysis in a non-growing cell suspension, causing a doubling of the rate of release of radioactively labelled wall material . Heat-inactivated cells did not autolyse and were not lysed by lysozyme unless they were supplemented by unheated cells or cell-free autolysate . Enhancement of autolysin activity could also be effected by pre-treatment of heated cells with lysozyme . The action of lysozyme on isolated cell walls released some free reducing groups, indicating limited breakage of the polysaccharide chains of peptidoglycan, and it was concluded that lysozyme modified the peptidoglycan and made it more susceptible to autolysin(s) . Lysozyme also enhanced the rate of septum separation and the probable significance of the results in relation to the control of cell separation is discussed. Am J Pathol, 1979 Nov, 97(2), 235 - 46 Chronologic changes of activities of naphthol AS-D acetate esterase and other nonspecific esterases in the mononuclear phagocytes of tuberculous lesions; Tsuda T et al.; Nonspecific esterases of mononuclear phagocytes (MNs) were studied histochemically in the developing and healing tuberculous lesions produced in rabbit skin by bacille Calmette Guerin (BCG) . Nonspecific esterases were assayed with the following substrates: naphthol AS-D acetate (AS-D), naphthol AS-D chloroacetate (AS-D Chl), naphthol AS acetate (AS) and alpha-naphthyl acetate (alpha-N), beta-Galactosidase, a lysosomal enzyme of MNs, was also assayed as a marker of MN activation . The number of MNs hydrolyzing AS-D Chl, AS, and alpha-N increased for 2 to 4 weeks after infection . These chronologic changes were similar to that of beta-galactosidase . In contrast, MNs hydrolyzing AS-D appeared predominantly in the healing lesions five to six weeks after infection . These MNs had the morphologic features of balloon-like cells . They contained few lysosomes and gathered in clumps far from the caseous center . The activity of the AS-D esterase was almost completely inhibited by various trypsin inhibitors, but not by the serine esterase inhibitor of phenylmethylsulfonyl-fluoride . These results suggest that the AS-D esterase is a trypsin-like esterase which participates in the healing of tuberculous lesions. Proc Natl Acad Sci U S A, 1979 Nov, 76(11), 5882 - 5 A practicable immunological approach to block spermatogenesis without loss of androgens; Talwar GP et al.; The intrinsic capacity of the immune system to elicit immune response selectively against late developing sperm proteins has been mobilized to intercept spermatogenesis . Bacillus Calmette-Guerin given in appropriate doses intratesticularly is effective in bringing about this effect . In dogs and rhesus monkeys, the sperm count in the semen declined precipitously, and almost complete azospermia was attained in 4-6 weeks after immunization . The few sperm cells that were present were immotile . Examination of serial sections of testes in immunized rats showed about 98% of the tubules to be devoid of sperm . The tubules were partially or fully atrophied . The basement membrane was, however, intact and the pertubular cell layer was normal . Sertoli cell nuclei were apparently normal but the cytoplasm was vacuolated and, in most cells, partially disintegrated . The lumen of the tubules was exhausted of formed elements and at times filled with eosinophilic debris . Leydig cells were present and hyperplasia of interstitial cells was seen, with massive infiltration of leukocytes . Blood testosterone levels were in the normal range and Leydig cells were responsive to gonadotropins . Libido was intact . The method was applicable to a variety of mammalian species . The implications of the results are discussed. Cancer Res, 1979 Nov, 39(11), 4756 - 9 Enhancement of endotoxic shock by N-acetylmuramyl-L-alanyl-(L-seryl)-D-isoglutamine (muramyl dipeptide); Ribi EE et al.; We described elsewhere that the synergistic antitumor activity of endotoxic extracts from Re mutants of gram-negative bacteria and trehalose mycolate against guinea pig syngeneic line 10 tumor was abrogated after peptide substances accompanying these extracts had been removed . This activity could be restored by combining peptide-free endotoxin either with cell wall skeleton from Bacillus Calmette-Guerin, a polymeric mycolic acid-arabinogalactan-mucopeptide complex, or with a combination of two separate components, trehalose dimycolate and N-acetylmuramyl-L-alanyl-(L-seryl)-D-isoglutamine (MDP) . We report here that when a combination of endotoxin (150 microgram) and a mixture of MDP (150 microgram) and trehalose dimycolate (150 microgram) was inoculated into established dermal tumors, a significant number of the animals died, presumably of endotoxic shock . All surviving animals suffered severe but temporary lethargy . When administered alone intradermally in the dose levels tested, none of the components caused severe lethargy or lethality . The lethal effects of 159 microgram of MDP also occurred in combination with relatively weak endotoxic products, such as Pseudomonas vaccine (Pseudogen), and these effects did not depend upon the presence of malignant tissue . Guinea pigs inoculated i.v . were even more susceptible inasmuch as the addition of as little as 6 microgram of MDP to 150 microgram of Pseudogen, itself not lethal, caused the death of 80% of the animals. Nucleic Acids Res, 1979 Oct 25, 7(4), 997 - 1010 Structure of cloned ribosomal DNA cistrons from Bacillus thuringiensis; Klier AF et al.; A library of B . thuringiensis DNA has been prepared by using the plasmid pBR322 as a cloning vehicle and E . coli as a host cell . By screening this collection with specific probes, 17 clones were identified whose hybrid plasmids contain rRNA genes of B . thuringiensis . Several of these plasmids have been mapped with restriction endonucleases and by DNA-RNA hybridization . By using maps of overlapping fragments, we have been able to establish an overall map of the ribosomal gene cluster. Biochemistry, 1979 Oct 16, 18(21), 4532 - 6 Comparison of the effect of linear gramicidin analogues on bacterial sporulation, membrane permeability, and ribonucleic acid polymerase; Paulus H et al.; Various analogues of linear gramicidin were tested for their biological activity in restoring the normal spore phenotype of gramicidin-negative mutants of Bacillus brevis and for their ability to increase cation conductivity of black lipid membranes and to inhibit bacterial RNA polymerase . Whereas many biologically active gramicidin analogues had no effect on membrane permeability, all biologically active peptides were able to inhibit ribonucleic acid (RNA) polymerase . These observations make it unlikely that membranes are the site of action of gramicidin during bacterial sporulation, but they are consistent with the notion that gramicidin functions to control RNA synthesis during the transition from vegetative growth to sporulation (Sarkar & Paulus, 1972) . The relationship between peptide structure and the ability to restore normal sporulation and inhibit RNA polymerase showed that the eight amino-terminal residues have little influence on the function of gramicidin, whereas the highly nonpolar repeating sequence D-leucyl-L-tryptophan is essential for biological activity and may represent the site of interaction with RNA polymerase. Am J Pathol, 1979 Oct, 97(1), 137 - 48 Prostaglandin biosynthesis in pulmonary macrophages; Hsueh W; Cultured rabbit alveolar macrophages, prelabeled with 14C-arachidonic acid (AA), released into the medium a trace amount of labeled prostaglandins (PG) as well as their precursor, AA . Phagocytosis of zymosan, heat-killed Staphylococcus, or bacille Calmette-Guerin (BCG) increased the AA and PG release to 2--2.5 times control values . The released PGs consisted of PGE2, D2, F2 alpha, and 6-keto F1 alpha . Phagocytosis of latex particles had no effect on PG release . Indomethacin inhibited release of PGs but did not affect AA release at low doses . Analysis of the cellular lipids showed that zymosan decreased the radioactive label in phosphatidylcholine (PC), but not in other phospholipids or neutral lipids, suggesting that PC is the main source of AA for PG synthesis in pulmonary macrophages . Cytochalasin B (CB) at phagocytosis-inhibiting doses or below, markedly increased PG synthesis by zymosan-treated macrophages . These data suggest that PG release is not dependent on engulfment of the particles . Phagocytosis of zymosan (but not latex) also resulted in the release of two lysosomal enzymes, acid phosphatase and beta-glucuronidase, which appeared temporally associated with the release of PGs (but not to phagocytosis) . Furthermore, CB augmented the zymosan-stimulated release of these enzymes at the same doses stimulating PG synthesis . However, indomethacin, at a dose completely inhibiting PG synthesis, failed to block lysosomal enzyme release . Thus, the coincidental release of PGs and lysosomal enzymes is not the result of a regulatory role of PGs in the release of lysosomal enzymes, but probably is the result of a common pathway of stimulation . (Am J Pathol 97:137--148, 1979). Eur J Biochem, 1979 Oct, 100(1), 301 - 8 Cysteinyl-tRNA synthetase from Bacillus stearothermophilus . A structural and functional monomer; Bruton CJ et al.; A procedure is described for the purification of cysteinyl-tRNA synthetase as a side product of a multi-enzyme isolation from Bacillus stearothermophilus . The native and denatured enzyme are both shown to have a molecular weight of 54000 by gel filtration and sodium dodecyl sulphate/polyacrylamide gel electrophoresis respectively . Fingerprinting and peptide counting indicate that the polypeptide chain has a nonrepeating primary structure . The enzyme has only one binding site for each of its substrates (cysteine, ATP and tRNACys) as judged by equilibrium dialysis, active-site titration and fluorescence quenching . No evidence for the dimerisation of the enzyme in the presence of these substrates could be found . We conclude that cysteinyl-tRNA synthetase, which is the smallest aminoacyl-tRNA synthetase yet described, is both structurally and functionally monomeric. Eur J Biochem, 1979 Oct, 100(1), 29 - 30 Purification and properties of alanine dehydrogenase from Bacillus sphaericus; Ohashima T et al.; 1 . The bacterial distribution of alanine dehydrogenase (L-alanine:NAD+ oxidoreductase, deaminating, EC 1.4.1.1) was investigated, and high activity was found in Bacillus species . The enzyme has been purified to homogeneity and crystallized from B . sphaericus (IFO 3525), in which the highest activity occurs . 2 . The enzyme has a molecular weight of about 230 000, and is composed of six identical subunits (Mr 38 000) . 3 . The enzyme acts almost specifically on L-alanine, but shows low amino-acceptor specificity; pyruvate and 2-oxobutyrate are the most preferable substrates, and 2-oxovalerate is also animated . The enzyme requires NAD+ as a cofactor, which cannot be replaced by NADP+ . 4 . The enzyme is stable over a wide pH range (pH 6.0--10.0), and shows maximum reactivity at approximately pH 10.5 and 9.0 for the deamination and amination reactions, respectively . 5 . Alanine dehydrogenase is inhibited significantly by HgCl2, p-chloromercuribenzoate and other metals, but none of purine and pyrimidine bases, nucleosides, nucleotides, flavine compounds and pyridoxal 5'-phosphate influence the activity . 6 . The reductive amination proceeds through a sequential ordered ternary-binary mechanism . NADH binds first to the enzyme followed by ammonia and pyruvate, and the products are released in the order of L-ALANINE AND NAD+ . The Michaelis constants are as follows: NADH (10 microM), ammonia (28.2 mM), pyruvate (1.7 mM), L-alanine (18.9 mM) and NAD+ (0.23 mM) . 7 . The pro-R hydrogen at C-4 of the reduced nicotinamide ring of NADH is exclusively transferred to pyruvate; the enzyme is A-stereospecific. Eur J Biochem, 1979 Oct, 100(1), 189 - 96 {Characterisation of a new endopeptidase from sporulating Bacillus sphaericus which is specific for the gamma-D-glutamyl-L-lysine and gamma-D-glutamyl-(L)meso-diaminopimelate linkages of peptidoglycan substrates (author's transl)}; Vacheron MJ et al.; A new peptidase which splits substrates related to the peptidic chains of peptidoglycans was found in the cell cytoplasm of sporulating Bacillus sphaericus . This is a gamma-D-glutamyl-L-diaminoacid endopeptidase (endopeptidase II) . It was shown to have substrate requirements different from those of the previously described gamma-D-glutamyl-(L)meso-diaminopimelate endopeptidase (endopeptidase I) . The substrates for endopeptidase II are peptides of the general type: formula: (see text) . Unsubstituted N-terminal L-alanine was a strict requirement for endopeptidase II activity . Specific activities were variable with the nature and the substitution of the diaminoacid C-terminal groups . The role of endopeptidase II in the biosynthesis of the spore cortex is discussed. J Antibiot (Tokyo), 1979 Oct, 32(10), 1011 - 5 Characterization of methanol extraction residue (MER) from Bacillus Calmette-Guérin (BCG); Shaikh B et al.; Analysis of methanol extraction residue (MER) from Bacillus Calmette-Guerin (BCG) was carried out to determine some specific chemical compositional characteristics . Samples of MER were found to contain approximately 40% protein and/or peptide, 3% soluble lipids, 17% bound lipids, 8% elemental nitrogen, and less than 2% mycolic acids . Amino acid analysis showed the presence of alanine, glycine and glutamic acid as the major amino acids . The data are reported in terms of the range found for each constituent over the samples analyzed . Somewhat consistent results were obtained between different MER preparations, but notable compositional variations were observed in samples of MER suspensions. Pathol Biol (Paris), 1979 Oct, 27(8), 491 - 2 {Influence of reintervention by the bacillus of Calmette and Guérin (BCG) on the nonspecific resistance of the mouse to Schistosoma mansoni (author's transl)}; Tribouley J et al.; Significant resistance to Schistosoma mansoni appears in mice two weeks after treatment with BCG . The state of nonspecific resistance is considerably increased if the injection of BCG two weeks prior to the infestation test is carried out using mice who have already received BCG . Thus, BCG innoculation in an organism presentized to the bacillus antigens increases immunostimulation . According to these results, during primoinnoculation with BCG, immunostimulation is not significant before this second week, that is, the time necessary for a state of hypersensitivity to appear . Nevertheless, it should be noted that weak yet detectable immunostimulation can be observed before the 14th day following a BCG primoinfection . Thus, the possible role of other mechanisms should be considered. J Gen Microbiol, 1979 Oct, 114(2), 483 - 6 The growth and form of bacterial colonies; Wimpenny JW; A simple method is described for measuring the profile of bacterial colonies . Profiles were determined for colonies of Bacillus cereus, Escherichia coli and Staphylococcus albus of different ages . In spite of differences in cell morphology, the colony profiles had a common basic structure consisting of steeply rising leading edge connected by a ridge to an interior region where height also rose, though less steeply, to a flat or domed centre . The colony mass increased exponentially through part of the growth phase . It is suggested that net colony growth consists of a combination of leading edge growth, which is unrestricted and approaches the maximum specific growth rate of the organism, and diffusion-limited growth in the colony interior . Common elements of profiles from each species may be a consequence of such differences in growth rate. J Antibiot (Tokyo), 1979 Oct, 32(10), 995 - 1001 Studies on bacterial cell wall inhibitors . VIII . Mode of action of a new antibiotic, azureomycin B, in Bacillus cereus T; Spiri-Nakagawa P et al.; Azureomycin B, a new antibiotic which contains sugar, amino acid and phenol moieties and inhibits Gram-positive bacteria, was found to be a specific inhibitor of peptidoglycan synthesis in bacteria . The antibiotic lysed growing cells of Bacillus cereus T at a concentration of 10 micrograms/ml but did not affect resting cells . Microscopical observation revealed swelling and lysis of the bacterial rods when treated with azureomycin B . The incorporation of {3H}diaminopimelic acid or {14C}glucosamine into acid-insoluble fraction of growing cells of Bacillus cereus T was strongly inhibited in the presence of azureomycin B, but that of {14C}leucine, {3H}thymidine or {3H}uridine were not, at least until 5 minutes after the beginning of the incubation . The antibiotic caused accumulation of a nucleotide precursor in the cells which was identified as UDP-MurNAc-L-Ala-D-Glu-meso-Dpm-D-Ala-D-Ala . Thus the site of action was suggested to lie between this nucleotide and peptidoglycan in the pathway of peptidoglycan synthesis. C R Seances Acad Sci D, 1979 Oct 1, 289(6), 549 - 52 {Toxicity of Bacillus thuringiensis var . israelensis for Simulium larvae, vectors of onchocerciasis}; Guillet P et al.; On field trials in infested streams, the primary powder R 153-78 made of B . thuringiensis var . israelensis with a potency of 3,000 I.T.U . Aedes aegypti/milligramme is very toxic for Blackfly larvae, especially for Simulium damnosum s.l . larvae . The lethal concentration 100, in 24 h . is 0.2 X 10(-6) for a 10 min . treatment . This high toxicity, specific for Diptera larvae such as Mosquitoes and Blackflies, is related to the special characteristic of the bacterial crystal protein which has a clear serological and chemical individuality compared to the crystal proteins of the other B . thuringiensis serotypes essentially pathogenic for Lepidoptera larvae . These preliminary results are very promising and could lead, on further research, to the utilization of B . thuringiensis var . israelensis on large scale for onchoceriasis vector control in Africa. J Infect Dis, 1979 Oct, 140(4), 541 - 5 Effect of bacille Calmette-Guérin on the immune response of BALB/c mice to a tumor allograft; Treagan L et al.; The effect of dosage and route of inoculation of bacille Calmette-Guerin (BCG) on immune response to allogeneic tumor cells was investigated . BALB/c mice were tested 14 and 21 days after injection of EL-4 lymphoma for spleen-cell cytotoxicity against EL-4 cells in vitro and for complement-dependent, antibody-mediated lysis of tumor cells . BCG treatment had no measurable effect on the antibody-mediated lysis of tumor cells, but spleen-cell cytotoxicity was significantly increased in mice treated with 10(4) or 10(8) BCG by the intraperitoneal route; no such increase occurred when BCG was given by the oral or subcutaneous routes . The cytotoxic effector cells were primarily thymus-derived, since treatment of spleens with rabbit antiserum to mouse brain serum decreased cytotoxicity titers by approximately 90% . Within the framework of these experiments, the intraperitoneal route of BCG inoculation resulted in a more effective immune stimulation than the oral or subcutaneous routes. Appl Environ Microbiol, 1979 Oct, 38(4), 656 - 8 Toxicity of parasporal crystals of Bacillus thuringiensis subsp . israelensis to mosquitoes; Tyrell DJ et al.; Toxicity of Bacillus thuringiensis subsp . israelensis (ONR-60A/WHO 1897) parasporal crystals to three medically important mosquito larvae is described . The numbers of larvae killed are in relation to crystal dry weight . The crystals are lethally toxic to Aedes aegypti Linnaeus (mean 50% lethal concentration {LC50} = 1.9 x 10(-4) micrograms/ml), Culex pipiens var . quinquefasciatus Say (LC50 = 3.7 x 10(-4) micrograms/ml), and Anopheles albimanus Wiedemann (LC50 = 8.0 x 10(-3) micrograms/ml) . Purfied crystals of B . thuringiensis subsp . kurstaki, which are toxic to lepidopteran insects, are ineffective against the mosquito larvae . Likewise, B . thuringiensis subsp . israelensis parasporal crystals are not efficacious for larvae of the lepidopteran, Manduca sexta. Carbohydr Res, 1979 Oct, 75, 243 - 50 Purification and physiocochemical properties of an extra-cellular cycloamylose (cyclodextrin) glucanotransferase from Bacillus macerans; Stavn A et al.; An extracellular cycloamylose (cyclodextrin) glucanotransferase (EC 2.4.1.19) from Bacillus macerans was purified to homogeneity by adsorption on starch, ammonium sulfate fractionation, column chromatography on DEAE-cellulose, and gel filtration on Sephadex G-100 . The enzyme had a molecular weight of 67,000 and consisted of one polypeptide chain . The isoelectric point was pH 5.4 . Temperature and pH optima were 60 degrees and 5.4--5.8, respectively . The purified enzyme was quite stable at 50 degrees (pH 6.0), but lost approximately 80% of its activity at 60 degrees for 30 min (pH 6.0) . Prolonged digestion by trypsin did not affect the catalytic properties of the enzyme . The Km for starch was 5.7 mg/ml. Biochim Biophys Acta, 1979 Sep 11, 547(3), 531 - 43 Events surrounding the early development of Euglena chloroplasts . 16 . Plastid thylakoid polypeptides during greening; Bingham S et al.; Using sulfolipid to locate plastid thylakoid membranes in gradients from dark-grown resting cells it has been possible to study the plastid thylakoid membrane polypeptides of Euglena gracilis var . bacillaris undergoing light-induced chloroplast development . All plastid thylakoid bands seen in dark-growing wild-type cells and in mutant W3BUL in which plastid DNA is undetectable, are observed to increase in amount during plastid development . Others, which are undetectable in dark-grown wild-type and W3BUL increase greatly during plastid development and appear to be those associated with pigment-protein complexes . The data obtained from experiments where the polypeptides were labeled with 35S during development, either continuously or in pulses, were consistent with these findings . Cycloheximide strongly inhibited the increases in amount in all bands and chloramphenicol or streptomycin produced a lower level of inhibition in all bands indicating tight control of theformation of each plastid membrane constituent by the others . The formation of a polypeptide band of 25 000 molecular weight, thought to be a part of a pigment-protein complex of the thylakoid, and chlorophyll synthesis were inhibited identically by these antibiotics. Biochim Biophys Acta, 1979 Sep 11, 547(3), 512 - 30 Events surrounding the early development of Euglena chloroplasts . 15 . Origin of plastid thylakoid polypeptides in wild-type and mutant cells; Bingham S et al.; Techniques are described for the isolation of plastid thylakoid membranes from light-grown and dark-grown cells of Euglena gracilis var . bacillaris, and from mutants affecting plastid development . These membranes, which have minimal contamination with other cell fractions, are localized in sucrose gradients by using the thylakoid membrane sulfolipid as a specific marker . The plastid thylakoid membrane polypeptides isolated from these membranes were separated on SDS polyacrylamide gels and yielded patterns containing 30-40 polypeptides . Light-grown strain Z gave patterns identical with bacillaris . Since the plastid thylakoid polypeptide patterns obtained from dark-grown wild-type cells and from a bleached mutant W3BUL in which plastid DNA is undetectable are identical, it appears that the proplastid thylakoid polypeptides of wild-type cannot be coded in plastid DNA and are probably coded in nuclear DNA . The plastid thylakoid polypeptide patterns obtained from various dark-grown mutants, making large but abnormal chloroplasts, show a correlation between the amount of chlorophyll formed and the amount of a plastid thylakoid polypeptide thought to be associated wtth one of the pigment-protein light-harvesting complexes . Treatment with SAN 9789 (4-chloro-5-(methylamino)-2(alpha, alpha, alpha,-trifluoro-m-tolyl)-3-(2H(pyridazinone) known to block carotenoid synthesis at the level of phytoene, causes a progressive loss of all plastid thylakoid polypeptides during growth in darkness and results in the establishment of a new, lowere steady-state level of sulfolipid . At least ten of the plastid thylakoid polypeptides become labeled when isolated chloroplasts are supplied with radioactive amono acids; of these six are undectable in W3BUL and are, therefore, candidates for coding by plastid DNA. J S Afr Vet Assoc, 1979 Sep, 50(3), 217 - 9 Determination of penicillin residues in milk - a comparison of two methods; Katz KW et al.; Two tests for penicillin residues in milk using the test organisms Sarcina lutea and Bacillus stearothermophilus var . calidolactis respectively are presented . The test methods are described, the results compared and the advantages of the latter test demonstrated. J S Afr Vet Assoc, 1979 Sep, 50(3), 151 - 3 The effect of the dye-marking of mastitis remedies on the incidence of antibiotic residues in Pretoria's market milk supplies; Bester BH et al.; A 1973 survey on the incidence of inhibitory substances (mostly antiobiotic residues) in market milk supplied in Pretoria, on 3 195 herd milk samples, 65 tanker milk samples and 252 samples of pasteurised milk using the disc assay procedure with Bacillus stearothermophilus C 953 as test organism, revealed inhibitory substances equivalent to 0,005 IU penicillin/ml in 7.8% of the herd samples, 29,2% of the tanker samples and in 38.5% of the samples of pasteurised milk . In 38.9% of the positive herd milk samples and 73% of the samples of pasteurised milk, penicillin was indentified with the aid of the penicillinase test . Some of the pasteurised milk contained inhibitory substances equivalent to more than 1.0 IU penicillin/ml; in some of the herd milk samples this figure exceeded 5,0 IU penicillin/ml . A repeat survey was undertaken in 1977/78 to evaluate the effect of compulsory dye-marking of non-prescription mastitis remedies on the situation . In a total of 1 081 herd milk samples, 60 tanker milk samples and 112 samples of pasteurised milk, antibiotic residues were found in 2,13% of the herd milk, 11,7% of the tanker milk and 2,1% of the pasteurised milk samples, with a much lower average concentration of antibiotic residues . The compulsory dye-marking of mastitis remedies had a beneficial effect on the occurrence of antibiotic residues in milk but did not ensure their complete absence, presumably because dye-marking was not made compulsory in scheduled preparations. Appl Environ Microbiol, 1979 Sep, 38(3), 379 - 84 Catabolism of 5-chlorosalicylate by a Bacillus isolated from the Mississippi River; Crawford RL et al.; A strain of Bacillus brevis isolated from a polluted section of the Mississippi River was shown to utilize 5-chloro-2-hydroxybenzoate (5-chlorosalicylate) as a sale source of carbon and energy . Enzymic analyses of cell-free extracts prepared from 5-chlorosalicylate-grown cells demonstrated that the initial step in the pathway involved cleavage of the aromatic ring between C1 and C2 by a specific 5-chlorosalicylate 1,2-dioxygenase . Loss of chloride from the growth substrate occurred after ring fission and was probably enzyme mediated . An intermediate chlorolactone apparently lost chloride by enzymatic hydrolysis with formation of maleylpyruvate . Maleylpyruvate was further degraded by both glutathione-dependent and glutathione-independent mechanisms, with these reactions being identical to the terminal reactions of the gentisate pathway . It was suggested that this novel 5-chlorosalicylate pathway may have evolved by recruitment of enzymes from an ancestral gentisate pathway. J Antibiot (Tokyo), 1979 Sep, 32(9), 952 - 8 Metabolic products of microorganisms . 184 . On the mode of action of cladosporin; Anke H; Cladosporin, a fungal isocoumarin derivative, strongly inhibits the uptake and thereby the incorporation of uracil and leucine into cells of Bacillus brevis and the incorporation of uridine but not leucine into cells of the ascitic form of Ehrlich carcinoma (ECA) of mice . Normal uptake was not restored by removal of the antibiotic . In cells of Escherichia coli A 19-15 (met-) the inhibition of methionine uptake is associated with the cessation of growth . In a methionine-prototrophic revertant from this organism, the uptake of methionine is still inhibited; growth, however, is hardly affected by cladosporin . In vitro no effect on the DNA-dependent RNA polymerase from E . coli and on the RNA polymerase II from wheat germ could be detected . The poly(U)-directed poly(Phe) synthesis was also not inhibited by cladosporin . It is concluded that cladosporin inhibits uptake processes which, for the case of essential nutrients, leads to loss of viability. Arch Esp Urol, 1979 Sep-Oct, 32(5), 417 - 26 {The BCG as coadjuvant in the treatment of bladder carcinoma}; Rodrigues Netto N Jr et al.; Thirty one patients with bladder carcinoma of different grades and stages were treated by endoscopic surgery and oral administration of bacillus Calmette-Guerin (BCG) . They were randomly separated in two groups: one was treated surgically and the other was treated surgically and with the BCG . According to the ability to develop an immune response to the antigens, patients were divided into three groups . The group who had been treated by endoscopic surgery only had 7 patients with recurrent bladder cancer during a median interval of 30 months . The group treated surgically and with BCG had only one patient with bladder cancer recurrence within a median follow-up period of 30 months . The results obtained with the use of BCG inducing tumor regression seem to indicate a favorable response. Antimicrob Agents Chemother, 1979 Sep, 16(3), 353 - 61 Correlation of aminoglycoside dosages with serum concentrations during therapy of serious gram-negative bacillary disease; Reymann MT et al.; We prospectively evaluated serum aminoglycoside (AMG) concentrations in 120 patients who received gentamicin or tobramycin for serious gram-negative bacillary disease . AMG serum concentrations were assayed by microbiological and radioimmunoassay techniques . Correlation between the two assay methods was good . When AMG doses were based on total body weight, there was no significant correlation between AMG dosage administered and serum concentrations in patients with either normal or abnormal renal function . The use of ideal body weight for calculation of AMG dosage improved this correlation significantly except in hemodialysis patients . AMG-induced nephrotoxicity occurred in 13 patients . No significant association was noted between the occurrence of toxicity and the specific AMG given or with other commonly recognized risk factors . Among study groups, peak AMG serum concentrations failed to exceed the minimal inhibitory concentration of the infecting organism in 17 to 33% of the cases . Serum inhibitory levels of greater than or equal to 1:8 were not associated with improved survival . There was no significant difference in mortality between the gentamicin- and tobramycin-treated groups . We advise base-line serum AMG levels in seriously ill patients with gram-negative bacillary disease and additional bacteriological studies in selected situations. Mikrobiologiia, 1979 Sep-Oct, 48(5), 880 - 6 {Electron microscopic study of the interaction between phages and Bacillus thuringiensis cells}; Smirnova TA et al.; The interaction of phages belonging to different morphological groups with the cells of Bacillus thuringiensis var . galleriae R and S variants was studied . No adsorption of phages Tg11 and Tg18 on the cells of R variant was found upon infection in a liquid medium . What is characteristic of phage Tg11 is that it is predominantly adsorbed at the poles of S variant cells . Phage Tg18 particles are uniformly distributed along the perimeter of S variant cells . Phage Tg13 is adsorbed on the both variant cells . Phage aggregates with the elements of cell walls having a tetrahonal assembly of the subunits can be revealed in phage Tg13 lysates . The size of the subunits is 7 nm and the distance between their centers is 11 nm . A structured element, apparently the T-layer, is involved in the adsorption of phage Tg13 on the cells. Am Rev Respir Dis, 1979 Sep, 120(3), 619 - 24 Various antigenic reactivities in delayed hypersensitivity among crystalline proteins from Mycobacterium phlei; Matsumoto J et al.; Comparisons were made of the delayed-type skin reactivity of 6 crystalline proteins purified from the cell extract of Mycobacterium phlei in guinea pigs sensitized with whole cells of the heat-killed bacillus . These highly purified proteins elicited varying degrees of cutaneous reaction . The most active protein had almost the same reactivity as purified protein derivative prepared from the culture filtrate of Mycobacterium phlei . On the other hand, the weakest protein did not elicit a marked cutaneous reaction even after injection of 3,000 times the amount of protein of the most potent one . The other 4 proteins showed moderate reactivities . The difference in antigenic potency between proteins is probably due not to the amounts of the proteins contained in the cells used for sensitization, but to their structure. Clin Orthop, 1979 Sep, (143), 15 - 29 A new look at the epidemiology of ankylosing spondylitis and related syndromes; Masi AT et al.; Among the rheumatic diseases, non so clearly illustrates the relations between host and environmental factors as the seronegative spondyloarthropathy group of disorders . The strongest association is with the histocompatibility antigen HLA-B27, which accounts for a striking susceptibility to these diseases and is present in over 90% of individuals with idiopathic ankylosing spondylitis . Next in importance appears to be a difference in sex penetrance with males predominating in all categories . The most dramatic sex relationship is with postvenereal Reiter's syndrome which has a male-to-female ratio of nearly 50:1 . Another potent host factor is age, with increased predisposition to onset at puberty and young adulthood in HLA-B27-positive patients . Environmental or possibly infectious agent influence are most apparent in Reiter's syndrome, where the antecedent circumstances of venereal contact and bacillary dysentery are frequent precipitating events . Secondary forms of peripheral arthritis, radiographic sacroiliitis, and ankylosing spondylitis frequently occur in psoriasis and inflammatory bowel disease; in the case of peripheral arthritis, there is no or a significantly reduced association with HLA-B27 compared to AS or RS . Secondary factor seem to be contributing to spondyloarthropathy in these disorders . These iterrelations emphasize the powerful effects of host characteristics on the type of rheumatic disease syndrome acquired and provide superb opportunities for more precise understanding of disease pathogenesis and ultimate control through the integration of epidemiologic, clinical, and laboratory research. Mikrobiologiia, 1979 Sep-Oct, 48(5), 868 - 72 {Structure and physico-chemical properties of bacterial flagella}; Biriuzova VI et al.; The structural organization of bacterial flagella was studied with Bacillus brevis and Escherichia coli MS 1350 . The presence of a spherical body at the basis of a flagellum was confirmed . The structural organization of ingredients of the flagellar appratus, its inner and outer part, was investigated . The molecular weight of protein subunits in the filamentous portion of the flagellum was assayed as well as their amino acid composition . The mode of attachment of the flagellum to the cell is discussed. Infect Immun, 1979 Sep, 25(3), 797 - 804 Inhibition by human serum of lymphocyte proliferation stimulated by purified protein derivative of tuberculin and bacillus Calmette-Guérin; Herman-Brand R et al.; The addition of normal human serum to murine lymphocyte cultures consistently depressed mitogen-induced transformation, as measured by deoxyribonucleic acid synthesis . Stimulation by the B-cell mitogens purified protein derivative-tuberculin, bacillus Calmette-Guerin, and lipopolysaccharide was consistently affected, but there was no inhibition of T-cells when human serum was added to concanavalin A-stimulated cultures . The inhibitory effects were not due to cytotoxic factors for B-lymphocytes or to specific antibodies in serum directed against the mitogens . Analogous results were found with guinea pig serum . Contact of the lymphocytes with the serum within the first 24 h of culture was necessary for inhibition. Cancer Res, 1979 Sep, 39(9), 3720 - 4 A comparative clinical and immunological assessment of methanol extraction residue of Bacillus Calmette-Guérin versus placebo in patients with advanced cancer; O'Connell MJ et al.; Eighty-four patients with advanced cancer refractory to conventional therapeutic modalities were randomly assigned in double-blind fashion to one of three intradermal treatment regimens: "high"-dose methanol extraction residue fraction of Bacillus Calmette-Guerin (MER) (2.0 mg); "low"-dose MER (0.5 mg); or 0.9% NaCl solution placebo . Toxicity, consisting primarily of cutaneous inflammation and ulceration, was limited to patients receiving MER and was most severe with the high-dose regimen . Pretreatment clinical and immunological parameters were comparable between patient groups . Although a significant number of patients had increases in various immune parameters according to the criteria used, there was no appearent advantage to MER given in either dosage schedule compared to placebo . Patient survival was not affected by either MER regimen compared to placebo . This investigation failed to demonstrate any significant clinical or immunological benefit from MER given in two dosage regimens in patients with advanced cancer with the laboratory methodology used and emphasizes the importance of appropriate controls in evaluating immunostimulants in humans. Cancer Res, 1979 Sep, 39(9), 3673 - 6 Toxicity of intrapleural Bacillus Calmette-Guérin treatment in animals; Filardi MJ et al.; The toxicity of intrapleural Tice strain Bacillus Calmette-Guerin (BCG) infection was tested in hamsters . Doses above 10(6) colony-forming units caused significant systemic infection, which could be controlled with conventional antituberculosis therapy . Living BCG in the pleural space did not prevent the healing of bronchial or vascular closures after pulmonary resection . Prophylactic intrapleural BCG (10(6) colony-forming units) significantly reduced tumor growth in the lungs of mice following i.v . injection of 5 x 10(5) syngeneic sarcoma cells . These animal experiments suggest that intrapleural BCG may be administered in the pleural space after lung resection in limited doses if followed by a complementary course of antimicrobial therapy. J Bacteriol, 1979 Sep, 139(3), 889 - 98 Regulation of phosphoglycerate phosphomutase in developing forespores and dormant and germinated spores of Bacillus megaterium by the level of free manganous ions; Singh RP et al.; The large depot of phosphoglyceric acid (PGA) which is accumulated within spores of Bacillus megaterium is greater than 99% 3-phosphoglyceric acid (3-PGA) . The 3-PGA depot is stable in forespores and dormant spores, but is utilized rapidly during spore germination . When spores were germinated in KBr plus NaF, the PGA depot was not utilized, but 13% of the 3-PGA was converted to 2-PGA . These data suggest phosphoglycerate phosphomutase as the enzyme which is regulated to allow 3-PGA accumulation during sporulation . Young isolated forespores, in which 3-PGA was normally stable, utilized their 3-PGA rapidly when incubated with Mn2+ plus the divalent cation ionophore X-537A; Mn2+ or ionophore alone or Mg2+ or Ca2+ plus ionophore was without effect . Young forespores contained significant amounts of Mn2+ . However, forespore Mn2+ exchanged slowly with exogenous Mn2+ and was removed poorly by toluene treatment . This suggests that much of the forespore Mn2+ is tightly bound to some forespore component . Since phosphoglycerate phosphomutase from B . megaterium has an absolute and specific requirement for Mn2+, these data suggest that the activity of this enzyme in vivo may be regulated to a large degree by the level of free Mn2+ . Indeed, the activity of this enzyme in forespore or dormant spore extracts was stimulated greater than 25-fold by Mn2+, whereas comparable extracts from cells or germinated spores were stimulated only two- to fourfold. Eur J Biochem, 1979 Sep, 99(3), 623 - 8 Effects of apramycin, a novel aminoglycoside antibiotic on bacterial protein synthesis; Perzynski S et al.; 1 . The novel aminoglycoside antibiotic apramycin is shown to be a potent inhibitor of protein synthesis in bacteria both in vivo and in vitro . 2 . In cell-free systems from Escherichia coli programmed with poly(U), apramycin induces translation errors, as assayed by incorporation of leucine, isoleucine and serine, although this effect occurs only to a limited extent . 3 . Apramycin inhibits the translocation step of protein synthesis both in vivo, in protoplasts of Bacillus megaterium, and in vitro, in cell-free systems from E . coli . It is proposed that this is the primary inhibitory effect of the drug. Zh Mikrobiol Epidemiol Immunobiol, 1979 Sep, (9), 41 - 5 {Immunologic study of the cellular components of bacillus pyocyaneus . V . Antigenic analysis of slime and its fractions}; Zaidner IG et al.; Various slime fractions were obtained from newly isolated mucoid strains of P . aeruginosa by the method of ultrafiltration or differential centrifugation with subsequent gel chromatography . Purified slime was found to react with a broader spectrum of typing O sera than the corresponding cell wall lipopolysaccharides . Erythrocytic diagnostic preparations produced on the basis of slime antigens allowed to reveale the presence of circulating antibodies against P . aeruginosa . The slime components with molecular weight of 30,000--100,000 daltons and greater contained common antigenic determinants, and the slime components with molecular weight of 10,000--30,000 daltons contained both specific antigenic determinants and those also common to the high molecular components. J Gen Microbiol, 1979 Sep, 114(1), 201 - 6 Nigericin-induced death of an acidophilic bacterium; Guffanti AA et al.; At an external pH of 3.5, nigericin (which catalyses an electroneutral H+/K+ exchange) abolished the transmembrane proton gradient (delta pH) of Bacillus acidocaldarius, causing a rapid acidification of the cytoplasm from approximately pH 6.0 to pH 3.5 . A pronounced loss of viability and fine-structural changes rapidly followed treatment with nigericin . A marked decline in respiration and an even more rapid decrease in cytoplasmic ATP were observed . Activity of at least one cytoplasmic enzyme decreased more slowly . There was no generalized loss in the integrity of the cytoplasmic membrane, as assayed by permeability to inulin or Na+ or by release of ultraviolet light-absorbing compounds . The loss of viability upon treatment with carbonyl cyanide m-chlorophenylhydrazone was similar to what observed with nigericin, so proton influx alone, rather than together with K+ efflux, was probably involved in the death of the organism . Moreover, acidification of the cytoplasm rather than abolition of the delta pH was the lethal event, since no loss of viability was observed when the delta pH was abolished by elevation of the external pH. Eur J Biochem, 1979 Aug 15, 99(1), 49 - 55 Biological role of gramicidin S in spore functions . Studies on gramicidin-S-negative mutants of Bacillus brevis ATCC9999; Marahiel MA et al.; Gramicidin-S-negative mutants of Bacillus brevis ATCC9999 have been isolated with a remarkly higher yield after ethidium bromide or acridine orange treatment, than after N-methyl-N'-nitro-N-nitrosoguanidine treatment . Four (MIV, Smr170, R5 and EB 16) of 38 isolated mutants were characterized with respect to the lesion in gramicidin-S-synthesizing activity . The mutants sporulate to the same extent as the parental strain except mutant Smr 170 which sporulates less . However, mutant spores were more heat-sensitive and possessed a reduced level of dipicolinic acid content . No significant difference was observed in the germination time of wild-type and mutant spores . All spores germinated after 80--110 min, but the outgrowth time was different: all gramicidin-S-negative mutants grew out immediately after germination whereas wild-type spores required a lag period of 9--10 h . When the mutants were allowed to sporulate in the presence of gramicidin S, the spores were found to be heat-resistant and their outgrowth postponed to the same period as the parent spores . The addition of gramicidin also eliminated the deficiency of dipicolinic acid . A new class of gramicidin-S-negative mutant, R5, which only activates L-valine and L-leucine, is described . A possible biological function of gramicidin S in the heat-resistance and in the timing of spore outgrowth is discussed. J Biol Chem, 1979 Aug 10, 254(15), 6838 - 41 Sequence-specific endonuclease Bgl I . Modification of lysine and arginine residues of the homogeneous enzyme; Lee YH et al.; The sequence-specific endonuclease Bgl I from Bacillus globigii (RUB561) has been purified to homogeneity as determined by denaturing polyacrylamide gel analysis . The active form of the enzyme is a single polypeptide with a molecular weight of 32,000 . The enzyme requires Mg2+ in the reaction mixture and displays a broad pH and monovalent cation requirement . Bgl I is not sensitive to sulfhydryl reagents but was affected by reagents that modify lysine and arginine residues . When lysine residues were modified by pyridoxal 5'-phosphate, both binding and catalysis were diminished while modification of arginine residues by 2,3-butanedione inhibited the enzyme activity but had no effect on its binding properties. J Biol Chem, 1979 Aug 10, 254(15), 6835 - 7 Identification of an NADH-linked disulfide reductase from Bacillus megaterium specific for disulfides containing pantethine 4',4''-diphosphate moieties; Swerdlow RD et al.; Bacillus megaterium contains an NADH-linked disulfide reductase that is specific for disulfides containing pantethine 4',4''-diphosphate moieties . This reductase is at its highest level in cells late in sporulation and in dormant spores, and could be involved in the formation and cleavage of coenzyme A-protein disulfides which take place late in sporulation and early in spore germination, respectively. Isr J Med Sci, 1979 Aug, 15(8), 668 - 74 Use of enzymatic and electron microscopy (freeze-etching) methods for studying ATP-dependent masking of erythrocyte membrane phospholipids; Loyter A et al.; Membrane phospholipids of ATP-depleted chicken, rat, and toad erythrocytes are more susceptible than fresh cells to hydrolysis by phospholipase C (Bacillus cereus), phospholipase A (bee venom), or the combination of these enzymes and sphingomyelinase . ATP depletion of chicken and rat erythrocytes greatly increased the membrane phospholipid fraction, which can be extracted by dry ether . Trinitrobenzene sulfonic acid attacked about 20% of the phosphatidylethanolamine of fresh chicken erythrocytes and 45% of ATP-depleted chicken erythrocytes . The intramembranous particles on the PF face of fresh chicken, rat, and toad erythrocytes are evenly distributed, while those in the same ATP-depleted erythrocytes are significantly clustered . The average distance between the intramembranous particles in the PF face of fresh chicken erythrocytes is about 13 nm, while in ATP-depleted erythrocytes it is about 30 nm . Studies with chicken erythrocytes revealed that ATP depletion is accompanied by dephosphorylation of certain membrane polypeptides . The correlation of dephosphorylation of membrane polypeptides, exposure of membrane phospholipids, and clustering of intramembranous particles is discussed. Am J Med, 1979 Aug, 67(2), 339 - 42 Legionnaires' disease . A cause of severe abscess-forming pneumonia; Lewin S et al.; In two previously well nonsmokers fatal pneumonia developed with extensive abscess formation . Legionnaires' bacillus was the only pathogen isolated . These cases indicate that Legionnaires' bacillus is capable of causing extensive necrosis of the lung. Cancer Res, 1979 Aug, 39(8), 3014 - 7 Effects of adriamycin and cyclophosphamide treatment on induction of macrophage cytotoxic function in mice; Stoychkov JN et al.; The effects of i.p . and s.c . Adriamycin and cyclophosphamide treatment of BALB/c x DBA/2F1 mice were studied alone and in combination with immunotherapeutic agents, pyran copolymer and Bacillus Calmette-Guerin, on macrophage cytotoxic ability, As assessed by direct viable cell counts of MBL-2 leukemia cells, both Adriamycin and cyclophosphamide produced growth-inhibitory macrophages . This function after s.c . cytostatic treatment peaked at Day 1 and decreased progressively, attaining normal control values by Day 6 . When adjuvants, such as pyran and B . Calmette-Guerin, were administered i.p . simultaneously with s.c . Adriamycin or cyclophosphamide, adjuvant-induced cytotoxic function was not markedly affected . A better knowledge of the influence of cytostatic agents alone or combined with immunoadjuvants on macrophage cytotoxic ability may be useful in designing more effective chemoimmunotherapy protocols. Biotechnol Bioeng, 1979 Aug, 21(8), 1345 - 59 Proteolytic denaturation and methods of improving the stability of glucose isomerase preparations; Adler D et al.; Evidence is provided in support of proteolytic denaturation of free and immobilized preparations of glucose isomerase from a Bacillus species . A number of methods to improve the stability with respect to proteolysis have been tested and their advantages as well as shortcomings are discussed . These methods include hollow-fiber treatment, gel permeation, thermal treatment, and addition of protease inhibitors . The half-life of the free and the cellulose acetate fiber-entrapped preparations of glucose isomerase can be significantly improved . For example, the hollow-fiber treatment can improve the half-life by an order of magnitude. J Antibiot (Tokyo), 1979 Aug, 32(8), 828 - 33 Antifungal activity upon Saccharomyces cerevisiae of iturin A, mycosubtilin, bacillomycin L and of their derivatives; inhibition of this antifungal activity by lipid antagonists; Besson F et al.; The antifungal activity of three antibiotics of the iturin group: iturin A, mycosubtilin, bacillomycin L and of eleven methylated and acetylated derivatives of these antibiotics was tested upon Saccharomyces cerevisiae . The lowest MIC values were found for natural antibiotics . The substitution of polar groups diminished the antifungal activity . Various lipids, sterols, fatty acids, fatty acid methyl esters and phospholipids were tested as inhibitors of the antifungal activity of iturin A, mycosubtilin and bacillomycin L . Cholesterol was the strongest inhibitor upon the three antibiotics; ergosterol, oleic acid and cis-vaccenic acid were less potent inhibitors . Among phospholipids, phosphatidyl choline inhibited bacillomycin L and iturin A while diphosphatidyl glycerol inhibited bacillomycin L and mycosubtilin . The inhibitory effect appeared to be dependent on the nature of both the hydrophilic group and the fatty acid part of phospholipids. Acta Neuropathol (Berl), 1979 Aug, 47(3), 183 - 8 Scanning electron microscopy of epiplexus macrophages responding to challenge by bacillus Calmette-Guerin; Merchant RE; The present investigation examined the morphological characteristics of epiplexus macrophages following a single intracisternal injection of the antigen, bacillus Calmette-Guerin (BCG) . Three days following injection of BCG (0.5 - 4.0 X 10(8) viable microorganisms), mongrel dogs were perfused with buffered aldehydes . The choroid plexus of the lateral and third ventricles was removed and routinely prepared for scanning or transmission electron microscopy . Choroid plexuses from normal animals (no BCG injection) were similarly prepared . Macrophages of normal animals possessed smooth cell surfaces with usually one to three cytoplasmic processes . Following BCG injection, a 10-fold increase in the epiplex macrophage population was observed . Furthermore, the majority of these cells presented an abundance of cell surface microappendages; including blebs, ruffles and microvilli . Cell processes and microvilli frequently mediated contacts between widely separated macrophages . These associations may play a role in the initiation and/or maintenance of the cellular immune response to BCG. Am J Pathol, 1979 Aug, 96(2), 595 - 610 The tumoricidal properties of inflammatory tissue macrophages and multinucleate giant cells; Poste G; Peritoneal exudate cells from C3H/HeN mice infected with bacille Calmette Guerin (BCG) and subcutaneous inflammatory macrophages from uninfected mice exhibit spontaneous cytotoxicity for tumor cells in vitro, but their tumoricidal activity can be increased by incubation in vitro with lymphokines released by mitogen- or antigen-stimulated lymphocytes . Inflammatory macrophages from these sites are only susceptible to activation in vitro by lymphokines for a short period (less than 4 days) following their initial emigration from the circulation to the site of inflammation . The expression of tumoricidal activity by activated macrophages is similarly short-lived (less than 4 days) . Once the tumoricidal state is lost it cannot be restored by further incubation with lymphokines in vitro . Fusion of macrophages to form multinucleate giant cells (MGCs) accompanies the loss of tumoricidal activity and the onset of resistance to activation by lymphokines, but the fusion process is not responsible for these changes, since unfused macrophages are similarly affected . Activation and acquisition of tumoricidal properties is confined to young macrophages recruited from the circulation during acute inflammation . Older macrophages and MGCs in chronic inflammatory lesions in which recruitment of new macrophages has ceased are nontumoricidal and are refractory to activation by lymphokines in vitro . These findings are discussed in relation to the efficiency of macrophage-mediated destruction of tumors in vivo and the amplification of macrophage antitumor activity by immunotherapeutic agents. Proc Natl Acad Sci U S A, 1979 Aug, 76(8), 3805 - 8 Characterization of the iron-sulfur centers in succinate dehydrogenase; Coles CJ et al.; Two techniques have been applied to the determination of the number and type (2-Fe, 4-Fe) of iron-sulfur centers in the iron-sulfur flavoprotein succinate dehydrogenase {succinate:(acceptor) oxidoreductase, EC 1.3.99.1} . One procedure uses p-CF3C6H4SH as an extrusion reagent and Fourier transform 19F nuclear magentic resonance as the method of detection and quantitation of extruded cores of these centers in the form of {Fe2S2(SRF)4}2- and {Fe4S4(SRF)4}2- (RF = p-C6H4CF3) . The second procedure, interprotein core transfer, involves thiol displacement of iron-sulfur cores followed by specific core transfer to the apoproteins of Bacillus polymyxa ferredoxin and adrenodoxin . Detection and quantitation are accomplished by electron paramagnetic resonance of reduced proteins at low temperatures . Both procedures clearly show that succinate dehydrogenase contains two dimeric (Fe2S2) and one tetrameric (Fe4S4) centers per mole of histidyl flavin, accounting for all eight nonheme iron and eight labile sulfur atoms found by chemical analysis . These results remove uncertainties created by the less than stoichiometric amounts of binuclear centers detected by electron paramagnetic resonance after dithionite reduction and provide secure characterization of the iron-sulfur centers in this enzyme. Cancer Res, 1979 Aug, 39(8), 3262 - 7 Nonspecific adjuvant immunotherapy of lung cancer with cell wall skeleton of Mycobacterium bovis Bacillus Calmette-Guérin; Yasumoto K et al.; Nonspecific adjuvant immunotherapy with Bacillus Calmette-Guerin cell wall skeleton (BCG-CWS) was given to 155 lung cancer patients . Clinical effects of the BCG-CWS treatment were estimated by comparing the survival of the BCG-CWS group with that of a historical control group on the basis of 4-year results . Significant prolongation of survival time has been observed in Clinical Stages II, III (M0) and III (M1) . However, most Stage III patients who were given the BCG-CWS treatment died of cancer itself after marked prolongation of survival time . An increase in complete cure rate has been expected only in Stages I and II . Surgicopathological staging was used in resected cases . Resected cases at any stage were sensitive to treatment with BCG-CWS . Histologically, all types of lung cancer including squamous cell carcinoma, adenocarcinoma, and anaplastic carcinoma were sensitive to treatment with BCG-CWS . Intrapleural administration of BCG-CWS to patients with malignant pleurisy was effective in controlling the pleural effusion and prolonging the survival time . No serious complication has been experienced in our study. Zh Mikrobiol Epidemiol Immunobiol, 1979 Aug, (8), 63 - 7 {Cell structure and the pathogenicity of Brucella at different stages of L transformation}; Tolmacheva TA et al.; On the basis of changes in the biological properties and morphology of Br . abortus culture under the action of penicillin 3 stages of L-transformation in Brucella were determined . The prevalence of first bacilliform and then typical L-cells and rapid reversion hampering the determination of virulence were characteristic of the initial stage (passages 1-4) . Typical L-cells with the wrinkled surface, deep depressions and holes as well as a decrease in virulence and slight pathomorphological changes in the organs of the infected animals were characteristics of the intermediate stage (passages 5-10) . Typical L-cells and amorphous masses, a further decrease in virulence, pathomorphological changes of toxic character (only after the injection of L-culture in large doses) were characteristic of the late stage (from passage 11 and further on) . At all stages of L-transformation Brucella cultures showed a high reproductive capacity, binary division, the formation of elementary bodies by budding both inside and on the surface of L-cells. Br J Surg, 1979 Aug, 66(8), 577 - 9 Two gas-gangrene-like infections due to Bacillus cereus; Fitzpatrick DJ et al.; Two cases of postoperative gas-gangrene-like infection due to Bacillus cereus are reported, drawing attention to the fact that Bacillus cereus, a common environmental bacterium, can occasionally give rise to severe post-operative infection . Characteristics of the organism related to the epidemiology and pathogenesis of such infections are discussed. J Bacteriol, 1979 Aug, 139(2), 486 - 94 Localization of low-molecular-weight basic proteins in Bacillus megaterium spores by cross-linking with ultraviolet light; Setlow B et al.; Two low-molecular-weight basic proteins, termed A and B proteins, comprise about 15% of the protein of dormant spores of Bacillus megaterium . Irradiation of intact dormant spores with ultraviolet light results in covalent cross-linking of the A and B proteins to other spore macromolecules . The cross-linked A and B proteins are precipitated by ethanol and can be solubilized by treatment with deoxyribonuclease (75%) or ribonuclease (25%) . Irradiation of complexes formed in vitro between deoxyribonucleic acid (DNA) or ribonucleic acid and a mixture of the low-molecular-weight basic proteins from spores also resulted in cross-linking of A and B proteins to nucleic acids . The dose-response curves for formation of covalent cross-links were similar for irradiation of both a protein-DNA complex in vitro and intact spores . However, if irradiation was carried out in vitro under conditions where DNA-protein complexes were disrupted, no covalent cross-links were formed . These data suggest that significant amounts of the low-molecular-weight basic proteins unique to bacterial spores are associated with spore DNA in vivo. J Biochem (Tokyo), 1979 Aug, 86(2), 447 - 52 Cross-linking with diimidates of glutamine synthetase from Bacillus stearothermophilus; Sekiguchi T et al.; Glutamine synthetase {EC 6.3.2.1} from Bacillus stearothermophilus was modified with diethyl malonimidate (DEM), dimethyl adipimidate (DMA), and dimethyl suberimidate (DMS) . DMA modified most epsilon-amino groups . On modification with DMA, formation of 3 to 4 cross-links/subunit resulted in a large increase in thermostability . The activity, allosteric properties and fluorescence spectrum of the enzyme were not changed on cross-linking . The SDS-polyacrylamide gel electrophoretic profiles of DEM-, DMA-, and DMS-modified enzymes suggested that the interaction berween six subunits in each of the two hexagonal rings of the protein are heterologous and are different from those between the piled subunits on different rings. Z Lebensm Unters Forsch, 1979 Jul 27, 169(1), 4 - 8 The amylase-producing microflora of semi-preserved canned sausages: identification of the bacteria and characterization of their amylases; Mitrica L et al.; Thirteen strains of amylase-producing bacteria were isolated from semi-preserved canned sausages and their ingredients . All belonged to the genus Bacillus, and could be separated into 4 different groups . Two groups were different strain of B . subtilis, one was B . amyloliquefaciens and the last was B . macerans . The identification of the different bacteria species was supported by disc gel electrophoresis of the supernatant culture fluid, after growth . The amylases were characterized with regard to temperature optimum, pH optimum and thermostability . Although some of the amylases appear to be quite thermostable, the only explanation for starch degradation in semi-preserved foods seems to be the amylase production from outgrowing spores which survived the heat treatment. Biochemistry, 1979 Jul 10, 18(14), 3045 - 50 Potentiometric titrations and oxidation-reduction potentials of manganese and copper-zinc superoxide dismutases; Lawrence GD et al.; Bovine erythrocyte superoxide dismutase and two manganese-containing superoxide dismutases have been reduced by the indirect coulometric titration method with methylviologen as the mediator-titrant . On the basis of the titration data the manganese-containing superoxide dismutases contain 1 g-atom of metal per mol of enzyme (dimer) . E0' = +0.31 V for the enzyme from Escherichia coli which exhibits a complicated pH dependence above neutral pH . The Bacillus stearothermophilus manganese-containing enzyme has an E0' = +0.26 V and delta Em/pH is 50 mV . Bovine erythrocyte superoxide dismutase exhibits anomalous behavior in the coulometric titration curves, which is indicative of two nonequivalent copper centers in the enzyme . Addition K3Fe(CN)6 or K2IrCl6 to the enzyme solution, prior to coulometric titration, indicates that these anions bind preferentially to one of the copper centers. J Biol Chem, 1979 Jul 10, 254(13), 5672 - 83 Purification and characterization of a carboxypeptidase-transpeptidase of Bacillus megaterium acting on the tetrapeptide moiety of the peptidoglycan; DasGupta H et al.; The enzyme carboxypeptidase-IIW of Bacillus megaterium incorporates free diaminopimelate into purified bacterial walls . This enzyme can be solubilized from toluene-treated cells by LiCl extraction and has now been purified 106-fold to one major band on polyacrylamide gel electrophoresis . The enzyme has an apparent molecular weight of approximately 60,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by Sephadex G-100 gel filtration . Carboxypeptidase-IIW requires divalent cations and thiol group(s) for optimal activity . Product analysis indicates that the enzyme can hydrolyze the terminal D-alanine from the tetrapeptide of the peptidoglycan or replace it with a variety of amino acids with D-asymmetric centers for transpeptidation . Substrate specificity studies reveal that the enzymatic activity depends on the presence of N-acetyl-D-glucosamine of the GlcNAc-MurNAc-tetrapeptide . This specificity of carboxypeptidase-IIW for the N-acetyl-D-glucosamine explains in part the affinity of the enzyme for the cell wall of B . megaterium . The enzyme is compared to the carboxypeptidases-transpeptidases of other organisms with the similarities and differences discussed. Can Med Assoc J, 1979 Jul 7, 121(1), 45 - 54 Oral administration of BCG as an adjuvant to surgical treatment of carcinoma of the bronchus; Miller AB et al.; A controlled clinical trial of the value of bacille Calmette--Guerin (BCG) vaccine given orally to patients with resectable carcinoma of the lung was conducted in 18 centres across Canada . A total of 308 patients were included in the analysis, 155 in the BCG group and 153 in the control group . The two groups were similar at the time of admission to the trial . BCG (120 mg) was given orally at weekly intervals for 1 month, every 2 weeks up to 3 months and then every 3 months until the total duration of therapy was 18 months . Over a 3- to 5-year follow-up period after the operation there was no difference in survival between the two groups, the proportion alive at 2 years being 61% in the BCG group and 58% in the control group . There was also no evidence of differences in the time to the detection of recurrent or metastatic disease or in the distribution of such disease . An analysis of prognostic factors confirmed the poor survival associated with histologically confirmed lymph node involvement . It may be concluded that no favourable effect from the oral administration of BCG was demonstrated. Lab Anim, 1979 Jul, 13(3), 257 - 61 The influence of Bacillus piliformis (Tyzzer) infections on the reliability of pharmacokinetic experiments in mice; Fries AS et al.; The half-lives of warfarin and trimethoprim were significantly longer in mice acutely infected with Bacillus piloformis and in mice which ad clinically recovered from previous experimental infection with the organism . The volume of distribution of trimethoprim but not of warfarin was significantly greater in infected mice than in controls . Body clearances of warfarin was significantly reduced in both disease states . For trimethoprim this parameter was only reduced in the acute state of the disease . The importance of careful control of Tyzzer's disease in laboratory animals for use in pharmacological research is stressed. J Antibiot (Tokyo), 1979 Jul, 32(7), 734 - 9 In vitro actions of some antibiotics on phospholipases; Sugatani J et al.; The effects of some antibiotics on activities of phospholipase A2, B and C were investigated in vitro . Tetracyclines, macrolides, chloramphenicol and carbenicillin inhibited the activity of Crotalus adamanteus phospholipase A2 towards phospholipids of egg-yolk emulsions . When the ability to inhibit the activity of Penicillium notatum phospholipase B towards mixed micelles of phosphatidylcholine and Triton X-100 was investigated, polymyxin B was found to be inhibitory while chloramphenicol and carbenicillin were found to stimulate the activity of the phospholipase . The activity of Bacillus cereus phospholipase C towards the mixed micelles was inhibited by bleomycin, oleandomycin and chloramphenicol. Chronobiologia, 1979 Jul-Sep, 6(3), 187 - 201 Circadian influence on the immunization of mice with live Bacillus Calmette-Guérin (BCG) and subsequent challenge with Ehrlich ascites carcinoma; Tsai TH et al.; Non-specific immunostimulation with Bacillus Calmette-Guerin (BCG) is of current interest in the treatment of cancer . The main objective of the series of experiments described in this paper was to evaluate the influence the host's circadian system has on a . the stimulation of the immune system with BCG and b . the subsequent efficiency of that stimulated immune system against the Ehrlich Ascites Carcinoma (EAC) . There was a circadian rhythm in the length of survival time in non-immunized mice challenged with the EAC . Mice receiving an EAC challenge during the middle of the light period survived significantly longer than those challenged with the EAC around the time of transition from dark to light . Mice immunized with BCG and challenged with EAC also demonstrated a circadian rhythm in the length of survival 30 days after EAC challenge with 86% survivors in the mice treated at 10(00) and 60% survivors in the mice treated at 07(00) . The same relationship was also observed 70 and 80 days after EAC challenge . Eighty days after EAC challenge, a circadian rhythm was apparent in the frequency of solid tumors at the site of the initial EAC injection . The highest incidence of solid tumors occurred at 13(00) . A circadian rhythm was found in the increase in body weight between the first and second BCG or saline injections . Rectal temperatures recorded on the 8th, 12th and 16th day after EAC challenge were characterized by circadian rhythmicity . In the mice without development of ascites, the peak temperature consistently occurred at 01(00) . In the mice with ascites there was a phase advance in the rectal temperature rhythm of 3 h so that the peak in the rhythm consistently occurred at 22(00) . In the mice with ascites a further finding was an increasing hypothermia as the ascites continued to develop; however, this hypothermia was not detectable during the time of the peak (10(00)) in the temperature rhythm . The mice which did not die by the 80th day after EAC challenge were challenged again with 5.0 x 10(6) EAC cells, and during the next 46 days circadian variations were observed in the numbers of mice which survived . Similar changes were observed during an additional 46 days after a third EAC challenge of 41.5 x 10(6) cells. Mikrobiologiia, 1979 Jul-Aug, 48(4), 716 - 22 {Bacteriocin-like factor in Bacillus thuringiensis}; Netyksa EM et al.; The paper describes Bacillus thuringiensis var . galleriae strains possessing a bacteriocin-like factor (factor K) which inhibits the cultures of a number of serotypes of Bac . thuringiensis . The K+ cultures exhibit no antibacterial effect on other gram-positive and gram-negative microorganisms . The maximal level of factor K is found by the 8--9th hour of cultivation . No correlation has been established between the strucutred elements in the preparations of K+ cultures and the antibacterial activity . Apparently, factor K can be transferred to other strains. Mikrobiologiia, 1979 Jul-Aug, 48(4), 681 - 8 {Ultrastructural organization of Sulfobacillus thermosulfidooxidans cells and spores}; Golovacheva RS; The ultrastructural organization of Sulfobacillus thermosulfidooxidans cells and spores was studied by electron microscopy using the techniques of negative contrasting and thin sections . The cell wall was found to be 20--25 nm thick and to consist of a rigid layer bound to the cytoplasmic membrane with crosspieces . A microcapsule about 20 nm thick having an ordered structure is closely associated with the cell wall . A layer of slime which can reach 0.2--0.3 mcm and more in thickness is located at the outside . The flagellar apparatus is absent . The membranous apparatus has a peculiar structure . On thin cross-sections of intact cells, the cytoplasmic membrane profile is stained sharply asymmetrically . Numerous membrane invaginates located in the cytoplasm are morphologically simple vesicular formations and structures of a rather complicated unusual configuration . Lamella-like structures are also encountered, which divide, as it were, the cytoplasm into compartments . The contours of the intracellular membranes look thicker as compared with the cytoplasmic membrane . Small vesicular formations are occasionally encountered in the periplasm . Vast areas with inclusions of unknown chemical nature having a fine granular structure are located in the cytoplasm . The process of sporogenesis comprises the same stages as in Bacillus, and spores have a strucutre typical of the endospores. J Bacteriol, 1979 Jul, 139(1), 126 - 31 Effect of alanine-containing dipeptides on germination of Bacillus thiaminolyticus spores; Watabe K et al.; Stereoisomeric alanylalanine (Ala-Ala) derivatives were examined for their effects on germination of Bacillus thiaminolyticus spores . L-Ala-L-Ala and L-Ala-glycine were effective in inducing germination, and their activities were completely inhibited by D-Ala . L-Ala-D-Ala and glycine-D-Ala competitively prevented L-Ala-induced germination . Sarcosine- or beta-Ala-containing L-alanyldipeptides and eight kinds of alanyltripeptides did not show any detectable effect on germinability or any inhibitory effect . No detectable amounts of Ala were found in germination exudates when alanylpeptides were incubated with spores . The ability of these peptides to induce or inhibit germination depends on their steric conformation and a certain distance between the primary amino group and the free carboxyl groups . Involvement of L-Ala dehydrogenase in the initiation of germination is unlikely because L-Ala-L-Ala was not a substrate and L-Ala-D-Ala was not an effective inhibitor of enzyme activity. Surg Gynecol Obstet, 1979 Jul, 149(1), 17 - 21 Treatment for melanoma of the lower extremity with intralesional injection of bacille Calmette Guérin and hyperthermic perfusion; Storm FK et al.; Twenty-seven patients with locally recurrent melanoma of the lower extremity were treated during a six year period . Recurrences were noted with primary lesions, at Clark's level II, III, IV or V, greater than 1.7 millimeters in depth of invasion and included intransit metastases, satellitosis, subcutaneous metastases and combinations of the three . Median time to recurrence was 12 to 14 months whether or not lymphadenectomy had been performed or lymph node metastases were present . Local recurrence was treated initially with intratumor bacille Calmette Guerin; 20 of 27 patients had complete or transient disease control and 14 patients were alive at 1.5 to 55.0 months . More responders reacted to dinitrochlorobenzene and purified protein derivative skin tests, although these parameters did not predict response to therapy . If intratumor bacille Calmette Guerin therapy did not control local disease or if the disease progressed toward the upper third of the thigh, patients underwent hyperthermic perfusion with L-phenylalanine mustard . Nine patients underwent ten therapeutic perfusions with objective response in seven of nine, with five being alive at two to 65 months . Intratumor bacille Calmette Guerin therapy and subsequent hyperthermic perfusion in bacille Calmette Guerin failures are rational treatment alternatives for locally recurrent melanomas of the extremity. Cancer Res, 1979 Jul, 39(7 Pt 1), 2544 - 6 Bacillus Calmette-Guérin enhancement of colony-stimulating activity and myeloid colony formation following administration of cyclophosphamide; Ladisch S et al.; Pretreatment of mice with Bacillus Calmette-Guerin (BCG) enhances recovery of the peripheral granulocyte count from cyclophosphamide (CTX)-induced leukopenia . In the present study, in vitro bone marrow culture was used to assess the effect of BCG pretreatment on serum levels of myeloid colony-stimulating activity and on regeneration of bone marrow myeloid colony-forming units (CFU-c) following CTX . C57BL/6 mice received either BCG or 0.9% NaCl solution i.p . 8 days prior to the administration of CTX (300 mg/kg i.p.) . Following CTX, peak serum levels of myeloid colony-stimulating activity were strikingly higher in the BCG-pretreated mice {1320 +/- 30 (S.E.) units} than in the 0.9% NaCl solution-pretreated mice (764 +/- 37 units) . BCG pretreatment also resulted in higher numbers of marrow CFU-c during recovery from CTX (e.g., 43.9 +/- 1.8 versus 20.0 +/- 0.9 myeloid colonies/10(5) marrow cells, Day 3 post-CTX) . However, neither the rate of decline nor the absolute nadir of CFU-c, nor CFU-c cell cycle characteristics were affected by the pretreatment . The initial effect of i.p . BCG pretreatment on recovery from CTX-induced granulocytopenia is an augmentation of serum myeloid colony-stimulating activity which precedes the enhanced regeneration of CFU-c and the accelerated recovery of the peripheral granulocyte count. Zentralbl Bakteriol {Orig A}, 1979 Jul, 244(2-3), 275 - 84 The bacillus cereus toxin: isolation of permeability factor; Ezepchuk Yu V et al.; The Bacillus cereus protein has been obtained from culture fluid in homogenic form as indicated by SDS-disc electrophoresis and immunodiffusion not described before . The protein has a molecular weight of 100000 daltons . Purification was accomplished by the following steps: (1) removal of ballast nitrous components with DE-32 cellulose at pH 7.2; (2) removal of the proteins from the culture filtrate (deluted four times by water) with DE-32 cellulose at pH 8.6; (3) elution by 0.005 M tris-HCl buffer at pH 7.0 containing 0.5 M NaCl; (4) column rechromatography on DE-32 cellulose at pH 8.6 . The isolated protein was identified as a vascular permeability factor acording to the bluing zone in rabbit skin tests or to the bluing lung tissue in mice. Eur J Biochem, 1979 Jul, 98(1), 261 - 5 Binding of thiostrepton to a complex of 23-S rRNA with ribosomal protein L11; Thompson J et al.; Thiostrepton binds with high affinity and with a 1 : 1 stoichiometry to a complex formed between Escherichia coli 23-S ribosomal RNA and ribosomal protein L11 of E . coli or the homologous protein BM-L11 of Bacillus megaterium . In the presence of T1 ribonuclease, protein BM-L11 and thiostrepton protect from degradation a fragment of E . coli 23-S RNA estimated to be about 50 nucleotides in length. Eur J Biochem, 1979 Jul, 97(2), 541 - 5 Methylation of basic proteins in ribosomes from wild-type and thiostrepton-resistant strains of Bacillus megaterium and their electrophoretic analysis; Cannon M et al.; Ribosomes, radioactively labelled in vivo with both {1-14C}methionine and {methyl-3H}methionine, have been isolated from both wild-type and thiostrepton-resistant strains of Bacillus megaterium and their constituent proteins separated by two-dimensional gel electrophoresis . Ribosomes from the wild-type strain possess one basic protein that is extensively methylated . In contrast no such protein can be detected in ribosomes from the thiostrepton-resistant strain. Dig Dis Sci, 1979 Jul, 24(7), 560 - 4 Rod-shaped organism in the liver of a patient with Whipple's disease; Viteri AL et al.; Histological review of a liver biopsy from a patient with known Whipple's disease revealed a prominence of Kupffer cells containing PAS-positive granules . Electron microscopy revealed rod-shaped organisms in the Kupffer cells but the presence of these structures were not associated with overt liver injury . This is thought to be the first reported demonstration of these bacillary bodies in this location. J Biol Chem, 1979 Jun 10, 254(11), 4863 - 75 Cleavage of a COOH-terminal hydrophobic region from D-alanine carboxypeptidase, a penicillin-sensitive bacterial membrane enzyme . Characterization of active, water-soluble fragments; Waxman DJ et al.; D-Alanine carboxypeptidase (CPase), a detergent-soluble penicillin-sensitive membrane enzyme of Bacillus stearothermophilus, Mr = 46,500, was digested with either trypsin or alpha-chymotrypsin to yield water-soluble fragments, designated T-CPase and Chy-CPase, respectively, each of Mr = approximately 45,000 . These fragments were generated and purified in milligram quantities by digestion of CPase covalently immobilized on a penicillin affinity column . They retained full enzymatic activity, became significantly more resistant to thermal inactivation, and lost micellar detergent binding upon proteolysis . Each was derived from CPase by loss of a COOH-terminal hydrophobic peptide . CPase was reconstituted into bacterial lipid vesicles in an enzymatically active form . Penicillin-binding sites were equally distributed on both sides of the lipid bilayer, suggesting a random orientation of the CPase molecules . Neither T-CPase nor Chy-CPase reconstituted into lipid vesicles when treated in an identical manner . CPase was slowly cleaved from the surface of these vesicles by either trypsin or alpha-chymotrypsin, yielding T-CPase and Chy-CPase, respectively . These results demonstrate that CPase is comprised of a water-soluble catalytic domain and a COOH-terminal hydrophobic region which mediates the anchoring of this enzyme to the bacterial membrane. Nature, 1979 Jun 7, 279(5713), 500 - 4 Structure and control of phosphofructokinase from Bacillus stearothermophilus; Evans PR et al.; The allosteric enzyme phosphofructokinase binds its substrate fructose-6-phosphate between two subunits of the tetramer, and allosteric effectors between another pair of subunits . The effector binding site accommodates both the activator and the inhibitor . The substrate cooperativity and allosteric control are mediated by these ligand bridges between subunits. Biochim Biophys Acta, 1979 Jun 6, 568(2), 395 - 407 The solubilisation of the membrane-bound D-alanyl-D-alanine carboxypeptidase of Bacillus coagulans NCIB 9365; McArthur HA et al.; Protoplast membranes and the particulate D,D-carboxypeptidase of Bacillus coagulans NCIB 9365 were extremely resistant to disruption by either detergents or urea . A combination of urea and the non-ionic detergent Genapol X-100 was required to achieve a significant solubilisation of membrane protein and D,D-carboxypeptidase in an active form; the pH optimum for this treatment was pH 7.5 . Solubilisation of the enzyme was accompanied by a two-fold enhancement of activity . Kinetic results indicated that the enhancement may be due to an alteration in the conformation of the enzyme following disruption of membrane structure. Zentralbl Bakteriol {B}, 1979 Jun, 168(5-6), 538 - 45 Study on the possibility of adopting a complex method in Blattella germanica (L.) control on board ships; Ulewicz K et al.; Campaings to control the German cockroach, Blattela germanica, were conducted at 14-day intervals over a period of 5.5 months on two merchant vessels of similar size and type . On one ship only the chemical "Gertoks", carbamate insecticide containing 1% propoxur was applied, and on the second ship two preparations simultaneously, a chemical and a biological--"Biotrol 25 W" containing Bacillus thuringiensis . - In order to assess the success of the control, the extent of infestation in the accomodation was determined immediately before and 24 h after the spraying . - The susceptibility of the cockroaches caught on the ships, to the two preparations applied, was also determined . It was found that "Gertoks" was an efficient preparation, the biopreparation "Biotrol 25 W" being less efficient . - A certain increase in resistance of the cockroaches to both preparations, was noted . - The above observations indicate that a considerable drop in numbers of population of cockroaches can be achieved if the ship's crew carry out systematic thorough spraying of ships quarters infested by cockroaches, with Gertoks" . They also confirm previous suggestions that complex method in B . germanica control on board ships is possible, using chemical and biological preparations containing Bac . thuringiensis spores simultaneously. Appl Environ Microbiol, 1979 Jun, 37(6), 1113 - 7 Comparison of the efficacy of steam sterilization indicators; Lee CH et al.; Twenty-one commercially available chemical steam sterilization indicators were processed in an empty autoclave for various times at temperatures between 240 and 270 degrees F (ca . 116 and 132 degrees C) . The time required to reach a sterilized reading at each temperature was plotted on a semilogarithmic time-temperature plot and compared with the time-temperature sterilization curve for Bacillus stearothermophilus . Five of the indicators had time-temperature kinetics similar to those of B . stearothermophilus, but three of these overestimated the effect of processing . Two of the indicators overestimated the effect of processing and were less sensitive to temperature changes when was B . stearothermophilus . Thirteen of the indicators had time-temperature curves that crossed the B . stearothermophilus plot . One indicator produced such ambiguous results that no determinations could be made with it . Out of 21 indicators tested, only 2 appear to be capable of accurately integrating the time-temperature effect at temperatures between 240 and 270 degrees F . The other indicators should be used only after careful analysis of their suitability for use at a given temperature. J Bacteriol, 1979 Jun, 138(3), 999 - 1009 Isolation, characterization, and in vitro assembly of the tetragonally arrayed layer of Bacillus sphaericus; Hastie AT et al.; The tetragonally arranged cell wall layer (T-layer) of Bacillus sphaericus NTCC 9602 was isolated and characterized . Parallel studies were made on a spontaneous variant of the wild-type strain which had a T-layer subunit of altered molecular weight . A purification method for the T-layers was devised which involved separation of the cell walls from the cytoplasmic contents, urea dissociation of the T-layer from the cell walls, removal of soluble contaminants by differential centrifugation, and finally selective adsorption of uncleaved subunits to sacculi . The purified subunits retained the capacity to form an assembly in vitro with the same lattice parameters as that observed on whole cells or cell walls and could readsorb to the cell walls from which they had been extracted . Both the wild-type and the variant subunits behaved as single, homogeneous polypeptide chains . Carbohydrate assay and isoelectric point determinations revealed that both subunit types were acidic glycoproteins . Values obtained for thebuoyant density, isoelectric point, and extinction coefficient differed minimally; major differences were observed in the molecular weight and the characterisitc width of cylinders formed by in vitro-assembled T-layer of the wild-type and variant . Assembled T-layer was subject to alkaline or acid dissociation and in acid titration dissociated at its isoelectric point. J Bacteriol, 1979 Jun, 138(3), 1010 - 21 Specific interaction of the tetragonally arrayed protein layer of Bacillus sphaericus with its peptidoglycan sacculus; Hastie AT et al.; Tetragonal layer protein (T-layer) isolated from Bacillus sphaericus NTCC 9602 (wild type) or 9602 Lmw (variant) bonded specifically to the sacculi (peptidoglycan) of either cell type . Only uncleaved T-layer subunits were capable of specific recognition of the B . sphaericus sacculi; other Bacillus strains and gram-positive bacterial sacculi would not adsorb B . sphaericus strain 9602 T-layer . The peptidogylcan did not function as a template since isolated T-layer subunits self-assembled into characteristic pattern . Upon reassociation with sacculi, T-layer assemblies were randomly oriented patches compared with more continuous strictly oriented pattern on cells or fresh cell walls . T-layer associated with the sacculus was less susceptible to conditions that dissociated in vitro-assembled T-layer . Mild proteolysis of both wild-type and variant T-layer subunits by a variety of enzymes reduced the molecular weight by 18,000 in all cases, indicating that one region of the molecule was particularly susceptible to cleavage . Subunits from which the minor fragment had been cleaved upon aging retained the capacity to assemble in vitro, but would no longer adsorb to sacculi . Thus, the ability of T-layer to form networks was separate from its ability to bind cell walls, and the 18,000-dalton piece of the T-layer polypeptide was necessary for attachment to the cell wall. Surg Gynecol Obstet, 1979 Jun, 148(6), 867 - 70 Intratumor bacillus Calmette-Guerin therapy for chest wall recurrence of carcinoma of the breast; Pardridge DH et al.; Intratumor bacillus Calmette-Guerin administration is effective for treating chest wall recurrence from carcinoma of the breast in selected patients . Our results suggest that bacillus Calmette-Guerin therapy can be used alone, with systemic chemotherapy, hormonal therapy or in previously irradiated tissue . It represents local treatment for local disease that is well tolerated . Intratumor bacillus Calmette-Guerin treatment is a useful clinical method for the treatment of local chest wall disease. Chest, 1979 Jun, 75(6), 685 - 7 Pulmonary manifestations in patients with malignant melanoma during BCG immunotherapy . A preliminary report; Bilgi C et al.; Ten patients with malignant melanoma who were treated with bacillus Calmette-Guerin (BCG) were studied with chest roentgenograms, scintiscans with radioactive 67gallium citrate, and the single-breath diffusing capacity of carbon monoxide (Dsb) . Three patients (stage 1B) with normal findings had no evidence of progression of disease . Seven patients had abnormal findings on at least one test . Two patients (stage 3B and stage 4B) with focal uptake in the lungs on the scintiscan had metastatic melanoma . Three patients (stage 3B) with diffuse uptake on the scintiscan and a reduced Dsb had no evidence of progression of disease . One patient (stage 1B) with an abnormal chest roentgenogram had a benign pleural effusion, and one patient (stage 4B) with a reduced Dsb was free of melanoma . A diffuse increase in the uptake of radioactive 67gallium in the lungs and a low Dsb may not indicate progression of melanoma . On the other hand, a focal uptake of 67gallium in the lungs is highly suggestive of metastatic disease. Int J Lepr Other Mycobact Dis, 1979 Jun, 47(2), 153 - 9 Lymphocyte transformation test in healthy contacts of patients with leprosy . II . Influence of consanguinity with the patient, sex, and age; Menzel S et al.; The study was carried out in the Gurage area of Ethiopia, where 53 household contacts of lepromatous patients, 37 household contacts of tuberculoid patients, and 91 control persons were examined with the lymphocyte transformation test (LTT) for their responses to whole and sonicated antigen preparation from M . leprae to BCG, M . avium, M . gordonae and phytohemagglutinin . The potential influence of host factors, namely the state of consanguinity with the leprosy patient, sex and age on the LTT responses was evaluated . In the 35 household contacts of "active," i.e., highly bacilliferous, lepromatous patients, consanguinity with a lepromatous patient was not associated with a significant depression of the LTT responses to M . leprae antigens . Male household contacts of active lepromatous patients showed significantly greater LTT responses to M . leprae antigens than female household contacts . Possible confounding factors for this finding are discussed . Sensitization of M . leprae antigens was present already in a high proportion of the 6 to 14 year old household contacts of active lepromatous patients, which was the youngest age group examined in our study . No significant results were found in any of the other patient contact groups with regard to the host factors examined. Int J Lepr Other Mycobact Dis, 1979 Jun, 47(2), 138 - 52 Lymphocyte transformation test in healthy contacts of patients with leprosy . I . Influence of exposure to leprosy within a household; Menzel S et al.; Fifty-three household contacts of lepromatous patients, 37 household contacts of tuberculoid patients, and 91 control persons were examined with the lymphocyte transformation test (LTT) for their responses to whole and sonicated antigen preparations from M . leprae, to BCG, M . avium, M . gordonae, and phytohemagglutinin (PHA) . The study was carried out in the Gurage area of Ethiopia in 15 households with a leprosy patient and 15 matched control households . Household contacts of lepromatous patients showed significantly greater LTT responses to antigens from M . leprae than the controls, whereas household contacts of tuberculoid patients did not respond differently from controls . Household contacts of lepromatous patients had significantly greater responses to M . leprae antigens when the index patients were "active," i.e., highly bacilliferous, than when they were "inactive," i.e., having a low bacillary load . The degree of sensitization, as indicated by the LTT response, in different exposure groups paralleled the degree of probable infectivity of the index patient . A preparation of antigen from whole M . leprae proved to be more sensitive and more specific in the LTT than did a sonicated preparation . A significant degree of cross-reactivity was found among the various mycobacteria in their LTT responses. Cancer Res, 1979 Jun, 39(6 Pt 1), 2125 - 31 Antibody-induced movement of common melanoma membrane antigens on the surface of unfixed human melanoma cells; Leong SP et al.; Antisera to common human melanoma antigens were obtained from melanoma patients receiving autologous immunization with their own irradiated cultured melanoma cells and Bacillus Calmette-Guerin . The antibody thus derived was used to detect common antigens on the plasma membrane of three different human melanoma cell lines by membrane immunofluorescence . The antigen-antibody complexes on the surface of melanoma cells would move to a pole (capping) and would subsequently be extruded into the extracellular milieu at room temperature . Approximately 25 to 30% of viable cells were positive by immunofluorescence . However, when the cells were fixed with methanol, 60 to 70% of cells demonstrated membrane binding . Capping was inhibited at 0 degrees or when the cells were pretreated with vinblastine sulfate . It can be concluded that common tumor antigens exist on the surface of viable human melanoma cells and that the redistribution of antigen-antibody complexes is an active process . The extrusion of antigen-antibody complexes in vitro may represent a mechanism of antigenic modulation in vivo and could indicate a basic method of tumor survival since presumably the antigen-denuded cell is viable and capable of replication but not of recognition by subsequent effector immune events. Biochem J, 1979 Jun 1, 179(3), 459 - 63 Identification of histidine residues that act as zinc ligands in beta-lactamase II by differential tritium exchange; Baldwin GS et al.; 1 . Four histidine-containing peptides have been isolated from a tryptic digest of the Zn2+-requiring beta-lactamase II from Bacillus cereus . One of these peptides probably contains two histidine residues . 2 . The presence of one equivalent of Zn2+ substantially decreases the rate of exchange of the C-2 proton in at least two and probably three of the histidine residues of these peptides for solvent 3H . 3 . It is concluded that peptides containing at least two of the three histidine residues acting as Zn2+ ligands at the tighter Zn2+-binding site of beta-lactamase II have been identified. Zentralbl Bakteriol {B}, 1979 Jun, 168(5-6), 525 - 32 {Disinfectant testing with Bacillus spores (author's transl)}; Bohm R et al.; This paper deals with special conditions for testing the sporocidal effect of disinfectants . A method for the production of spore suspensions (Fig . 1) and the negative influence of heating to its resistance against disinfectants was described (Fig . 2) . It was shown that spores of a non-virulent strain of Bac . anthracis could be used for this procedure (Fig . 3) . In performing the suspension--and the germ-carrier test it was necessary to watch the effect of postinhibition and of inactivators . The application of 10 microliter disposible loops in the suspension-test was recommended . Since the test for the evaluation of the influence of organic matter on the results of disinfection was unreliable (Fig . 4), it was proposed to replace it by invention of a safety-factor . The germ-carrier test should be done with constant amounts of spores, dried to the surface under defined conditions . Exposition of germ-carriers to disinfectants should be done in a chamber with constant temperature and rel . humidity, since the results are strongly influenced by these factors (Fig . 5). Biochem J, 1979 Jun 1, 179(3), 509 - 14 Unfolding and refolding of phospholipase C from Bacillus cereus in solutions of guanidinium chloride; Little C et al.; 1 . Protein-fluorescence studies indicated that phospholipase C from Bacillus cereus is denatured in solutions of guanidinium chloride . The denaturation was not thermodynamically reversible and followed biphasic kinetics . 2 . Guanidinium chloride solutions released the structural Zn2+ from the enzyme and rendered all histidine residues chemically reactive . In the presence of free Zn1+ the enzyme was much more resistant to denaturation . Also, the addition for free Zn2+ to the denatured enzyme induced refolding . 3 . The Zn2+-free apoenzyme was much more sensitive to guanidinium chloride than was the native enzyme and the denaturation appeared to be thermodynamically reversible . 4 . Guani