J Mol Biol, 1988 Apr 5, 200(3), 611 - 2 Crystallization of haloalkane dehalogenase from Xanthobacter autotrophicus GJ10; Rozeboom HJ et al.; Haloalkane dehalogenases are enzymes that release chloride or bromide from n-halogenated alkanes . X-ray quality crystals of haloalkane dehalogenase from the 1,2-dichloroethane-degrading bacterium Xanthobacter autotrophicus GJ10 have been grown at room temperature from 64% saturated ammonium sulfate solutions (pH 6.2 to 6.4) . The crystals diffract in the X-ray beam to at least 2.4 A resolution (1 A = 0.1 nm) . Their space group is P2(1)2(1)2, with cell dimensions a = 94.1 A, b = 72.8 A, c = 41.4 A and alpha = beta = gamma = 90 degrees . There is one monomer (molecular weight 36,000) per asymmetric unit. Biochemistry, 1988 Apr 5, 27(7), 2450 - 61 Cytochrome c and c2 binding dynamics and electron transfer with photosynthetic reaction center protein and other integral membrane redox proteins; Moser CC et al.; To further the understanding of the details of c-type cytochrome action as a redox carrier between major electron-transfer proteins, the single-turnover kinetics time course of cytochrome c and cytochrome c2 oxidation by flash-activated photosynthetic reaction center (purified from the bacterium Rhodobacter sphaeroides) has been examined under a wide variety of conditions of concentration, ionic strength, and viscosity with reaction center present in detergent dispersion and phosphatidylcholine proteoliposomes . We find that the three-state model proposed by Overfield and Wraight {Overfield, R . E., & Wraight, C . A . (1980) Biochemistry 19, 3322-3327} is generally sufficient to model the kinetics time course; many similarities are found with the cytochrome c-cytochrome c oxidase interaction in mitochondria . Further, we find the following: (1) Significant "product inhibition" by oxidized cytochrome c (c2) bound to the reaction center is apparent . (2) The viscosity sensitivity of the electron transfer into the reaction center from bound cytochrome c (c2) suggests a physical interpretation of the distal state . (3) The exchange dynamics of oxidized and reduced cytochrome c (c2) are similar regardless of the state of activation of the reaction center . (4) Preferential binding of the oxidized form of cytochrome c is revealed upon reconstitution of the reaction center into neutral lipid vesicles, permitting an independent confirmation of the binding suggested by the kinetics . (5) Flash-activated electron-transfer kinetics in reaction center hybrid protein systems have shown that diffusion and competitive binding characterize the behavior of cytochrome c as a redox carrier between the reaction center protein and either the cytochrome bc1 complex or the cytochrome c oxidase. Biochemistry, 1988 Apr 5, 27(7), 2444 - 50 A hypothetical model of the flavodoxin-tetraheme cytochrome c3 complex of sulfate-reducing bacteria; Stewart DE et al.; A hypothetical model of the flavodoxin-tetraheme cytochrome c3 electron-transfer complex from the sulfate-reducing bacterium Desulfovibrio vulgaris has been constructed by using interactive computer graphics based on electrostatic potential field calculations and previous NMR experiments . Features of the proposed complex are (1) van der Waals contact between the flavin mononucleotide prosthetic group of flavodoxin and one heme of the cytochrome, (2) unique complementarity of electrostatic fields between the region surrounding this heme and the region surrounding the exposed portion of the flavin mononucleotide group of flavodoxin, and (3) no steric interferences between the two polypeptide chains in the complex . This complex is consistent with all structural and spectroscopic data available. J Biol Chem, 1988 Apr 5, 263(10), 4602 - 6 Actinomycin synthesis in Streptomyces antibioticus . Purification and properties of a 3-hydroxyanthranilate 4-methyltransferase; Fawaz F et al.; A methyltransferase, which utilizes 3-hydroxyanthranilic acid (HAA) as a substrate, has been purified to near homogeneity from 30-36-h mycelium of the bacterium Streptomyces antibioticus . The enzyme was obtained in approximately 20% yield with a purification of 130-fold . Polyacrylamide gel electrophoresis under denaturing conditions indicates that the enzyme is composed of a single subunit with Mr of about 36,000 . On chromatography in 0.5 M NaCl, the enzyme displays a molecular weight of about 37,000 . The specific activity of the enzyme in S . antibioticus mycelium is maximal between 30 and 36 h following inoculation of galactose/glutamic acid medium and, at those times post-inoculation, the specific activity is essentially the same in extracts of mycelium obtained from cultures grown on glucose rather than galactose as the carbon source . The enzyme activity is stimulated by Na2EDTA (in crude extracts) and by 2-mercaptoethanol and the methyltransferase shows a strong preference for HAA as substrate as compared with a number of HAA analogs . Thin layer chromatography of ethyl acetate extracts of large-scale incubation mixtures confirms that the product of the reaction is 4-methyl-3-hydroxyanthranilic acid . The reaction product was also a substrate for phenoxazinone synthase and was incorporated into actinomycin by S . antibioticus mycelium . Kinetic parameters for the methyltransferase reaction was determined. Microbiologica, 1988 Apr, 11(2), 169 - 71 Isolation of Erysipelothrix rhusiopathiae from a 55 year old man and a positive tracing of the infection to chicken-raising premise; Salamah AA; Erysipelothrix rhusiopathiae was isolated from the swollen finger of a 55 year old man . The swelling was due to a peck by an infected chicken . Tracing the infection to the chicken-raising premises has proven that the bacterium was present in some of the chicken and manure samples. Int J Clin Pharmacol Ther Toxicol, 1988 Apr, 26(4), 199 - 203 A study of the levels of tobramycin and gentamicin in human serum and renal cortex and of changes in in vitro bacterial sensitivity; Farago E et al.; Concentration of gentamicin and tobramycin in sera and renal tissue was determined . Though between the 2nd and 6th day of the treatment, gentamicin cumulated in the renal tissue, but it did not reach toxic value . Tobramycin did not show cumulation tendency . According to in vitro bacterium sensitivity studies, the wide scale use of tobramycin in the clinical practice caused the sudden increase of resistance . To avoid this, the rules of antibiotic treatment should be observed. Appl Environ Microbiol, 1988 Apr, 54(4), 937 - 44 Mineralization of phenanthrene by a Mycobacterium sp; Guerin WF et al.; A Mycobacterium sp., designated strain BG1, able to utilize the polycyclic aromatic hydrocarbon phenanthrene as the sole carbon and energy source was isolated from estuarine sediment following enrichment with the hydrocarbon . Unlike other phenanthrene degraders, this bacterium degraded phenanthrene via 1-hydroxy-2-naphthoic acid without accumulating this or other aromatic intermediates, as shown by high-performance liquid chromatography . Degradation proceeded via meta cleavage of protocatechuic acid . Different nonionic surfactants (Tween compounds) solubilized the phenanthrene to different degrees and enhanced phenanthrene utilization . The order of enhancement, however, did not correlate perfectly with increased solubility, suggesting physiological as well as physicochemical effects of the surfactants . Plasmids of approximately 21, 58, and 77 megadaltons were detected in cells grown with phenanthrene but not in those which, after growth on nutrient media, lost the phenanthrene-degrading phenotype . Given that plasmid-mediated degradations of aromatic hydrocarbons generally occur via meta cleavages, it is of interest that the addition of pyruvate, a product of meta cleavage, supported rapid mineralization of phenanthrene in broth culture; succinate, a product of ortho cleavage, supported growth but completely repressed the utilization of phenanthrene . The involvement of plasmids may have given rise to the unusual degradation pattern that was observed. J Bacteriol, 1988 Apr, 170(4), 1709 - 14 Purification and properties of benzoate-coenzyme A ligase, a Rhodopseudomonas palustris enzyme involved in the anaerobic degradation of benzoate; Geissler JF et al.; A soluble benzoate-coenzyme A (CoA) ligase was purified from the phototrophic bacterium Rhodopseudomonas palustris . Synthesis of the enzyme was induced when cells were grown anaerobically in light with benzoate as the sole carbon source . Purification by chromatography successively on hydroxylapatite, phenyl-Sepharose, and hydroxylapatite yielded an electrophoretically homogeneous enzyme preparation with a specific activity of 25 mumol/min per mg of protein and a molecular weight of 60,000 . The purified enzyme was insensitive to oxygen and catalyzed the Mg2+ ATP-dependent formation of acyl-CoA from carboxylate and free reduced CoA, with high specificity for benzoate and 2-fluorobenzoate . Apparent Km values of 0.6 to 2 microM for benzoate, 2 to 3 microM for ATP, and 90 to 120 microM for reduced CoA were determined . The reaction product, benzoyl-CoA, was an effective inhibitor of the ligase reaction . The kinetic properties of the enzyme match the kinetics of substrate uptake by whole cells and confirm a role for benzoate-CoA ligase in maintaining entry of benzoate into cells as well as in catalyzing the first step in the anaerobic degradation of benzoate by R . palustris. J Bacteriol, 1988 Apr, 170(4), 1438 - 44 The methanoreductosome: a high-molecular-weight enzyme complex in the methanogenic bacterium strain Gö1 that contains components of the methylreductase system; Mayer F et al.; The methanogenic bacterium strain Go1 harbors a high-molecular-weight enzyme complex containing methyl coenzyme M methylreductase as revealed by immunoelectron microscopy . This complex consists of a spherelike, hollow head piece, in the wall of which a number of copies of the methyl coenzyme M methylreductase are located . It is named Rc (c indicates collector) . Intimately bound to it is a group of additional subunits of unknown composition referred to as Rm (m indicates mediator) . Electron microscopy of negatively stained samples indicated that Rm contains a functional pore or channel which connects the internal volume of Rc with the outside . The RcRm complex is named Rs (s indicates spherelike) . This complex was often found detached from the inside of the cytoplasmic membrane when membrane vesicles were investigated . However, Rs was also seen attached to a third component of the complex located in the membrane, the attachment being mediated by Rm . This membrane part of the complex is designated Rt (t indicates translocator) . It consists of subunits with unknown composition . When Rs is attached to the membrane, the pore in Rm appears to be plugged by Rt . This indicates that the internal volume in Rc is in contact, via the pore in Rm, with Rt . The RcRmRt complex is referred to as methanoreductosome . Functional implications of the structural organization of the methylreductase system are discussed in view of methane formation and the creation of a transmembrane proton gradient used by the cell for ATP synthesis. J Bacteriol, 1988 Apr, 170(4), 1843 - 7 Roles of bacteriochlorophyll and carotenoid synthesis in formation of intracytoplasmic membrane systems and pigment-protein complexes in an aerobic photosynthetic bacterium, Erythrobacter sp . strain OCh114; Iba K et al.; Synthesis of bacteriochlorophyll and carotenoids was inhibited in an aerobic photosynthetic bacterium, Erythrobacter sp . strain OCh114, by alpha, alpha'-dipyridyl and diphenylamine . Formation of two pigment-protein complexes, reaction center-B870 (RC-B870) and B806, and development of the intracytoplasmic membranes of the cells were studied by spectral analysis and electron microscopy . Inhibition of bacteriochlorophyll synthesis by alpha, alpha'-dipyridyl, which was accompanied by a decrease in carotenoid synthesis, suppressed formation of intracytoplasmic membranes in the cells . Growth under illumination had a similar effect on formation of pigments and membranes . On the other hand, inhibition of carotenoid synthesis by diphenylamine did not suppress either development of the membrane system or bacteriochlorophyll synthesis . Formation of RC-B870 and B806 complexes, however, was differentially affected by blockage of carotenoid synthesis . In the presence of diphenylamine, the B806 complex was formed in a much smaller amount than the RC-B870 complex . These results suggest that, in Erythrobacter sp . strain OCh114, bacteriochlorophyll plays an essential role in intracytoplasmic membrane development, and carotenoids are important for assembly of pigment-protein complexes. Biochimie, 1988 Apr, 70(4), 503 - 17 Strategies for the development of bacterial transformation systems; Mercenier A et al.; An effective transformation system is a prerequisite for facile genetic manipulation of bacteria . Bacteria may be naturally competent for transformation or may be treated with various agents, such as Tris buffers or divalent metal ions, to induce competence . Transformation can also be accomplished by electroporation, or by fusion of protoplasts with PEG in the presence of transforming DNA . Unfortunately, the mechanism by which cells become permeable to DNA and the process by which DNA enters the cells is frequently unknown . In order to establish a transformation system for an untransformable bacterium, recipient strains and transforming DNA must be carefully selected . Since it is impossible to predict in advance which method of transformation will be successful with a particular bacterial strain, several techniques are usually evaluated . This review describes a number of factors that appear to be critical for developing a transformation system and presents a strategy for experimentation with novel bacteria. J Antibiot (Tokyo), 1988 Apr, 41(4), 433 - 8 Novel macrocyclic antibiotics: megovalicins A, B, C, D, G and H . I . Screening of antibiotics-producing myxobacteria and production of megovalicins; Miyashiro S et al.; New antibiotics belonging to macrocyclic were discovered and named megovalicins A, B, C, D, G and H . The antibiotics were accumulated endogeneously by a newly isolated myxobacterium identified as Myxococcus flavescens . Tank culture of the bacterium for 4 days gave 8.8 g cells/liter on a wet basis, and 4.8, 7.1, 20.0, 0.4, 3.75 and 15.0 micrograms of megovalicins A, B, C, D, G and H respectively were obtained from 1 g wet cells. Indian J Lepr, 1988 Apr, 60(2), 159 - 72 Evaluation of an enzyme immunoassay based on sonicate supernatant antigens of Mycobacterium w for immunodiagnosis of leprosy; Moudgil KD et al.; An enzyme immunoassay (EIA) based on sonicate supernatant antigens of a cultivable, atypical bacterium, Mycobacterium w (M . w), for immunodiagnosis of leprosy is described . M . w was selected after screening of sonicate supernatant antigens of seven cultivable mycobacteria in EIA . The results of the assay were compared with that of EIA using phenolic glycolipid-I (PGL-I) . The M . w assay was more sensitive than PGL-I based EIA, for detection of leprosy patients of all categories, including long term treated patients with low bacterial load . The M . w assay was highly sensitive (93.49%) for detection of active LL patients, and the difference in the positivity of the two assays for LL patients was statistically significant (p 0.05) . The combined positivity of the assays with M . w and PGL-I for LL was higher than that with either antigen alone . M . w assay, in addition, was also highly sensitive for detection of patients with active pulmonary tuberculosis. J Biol Chem, 1988 Mar 25, 263(9), 4374 - 80 Surface-enhanced resonance Raman scattering spectroscopy of bacterial photosynthetic membranes . The carotenoid of Rhodospirillum rubrum; Picorel R et al.; Resonance Raman scattering by the carotenoid, spirilloxanthin (Spx), in a suspension of chromatophores (cytoplasmic side out) isolated from the photosynthetic bacterium, Rhodospirillum rubrum, is greatly enhanced when the membranes are adsorbed onto the surface of an anodized Ag electrode . The phenomenon is the basis for surface-enhanced resonance Raman scattering (SERRS) spectroscopy . The Spx SERRS peaks observed were at 1505-1510, 1150-1155, and 1000-1005 cm-1 with laser excitation wavelengths ranging between 457.9 and 568.2 nm . Similar peaks were not observed with spheroplasts (periplasmic side out) isolated from the same species . The difference in signal detected in chromatophores and spheroplasts is not due to differences in membrane surface charge, presence of residual cell wall on the spheroplast surface, lack of adhesion of spheroplasts to metals, or large differences in pigment content per unit membrane area . Instead, the results indicate an asymmetric distribution of Spx in vivo across the membrane (i.e., it is located on the cytoplasmic side of the membrane) . The results also demonstrate that the SERRS effect is extremely distance sensitive, and the thickness of a single bacterial membrane (separating the Ag electrode from the carotenoid) is sufficient to prevent detection of Spx spectra . Studies of chromatophores from the F24 strain (a reaction centerless mutant) have pin-pointed B880 antenna complex as the source of the Spx SERRS spectra, and a schematic model of the minimal structural unit of B880 is presented . This work demonstrates the potential of the SERRS technique as a probe for surface topology of pigmented membranes. Derm Beruf Umwelt, 1988 Mar-Apr, 36(2), 39 - 44 {The spectrum of Lyme borreliosis from the dermatologic viewpoint}; Aberer E et al.; Erythema chronicum migrans, lymphadenosis benigna cutis and acrodermatitis chronica atrophicans represent the dermatological manifestations of the multi-organ disease Lyme borreliosis . Koch's requirements of evidence for an infectious disease, demonstration of the bacterium, transfer, and culture have proven Borrelia burgdorferi to be the causative agent of the above mentioned skin diseases . This justifies a penicillin therapy, that has been administered in Europe empirically for the last 30 years . Correct and prompt diagnosis is important since delayed treatment is less effective, presumably because the spirochete becomes sequestered in immune-privileged sites . Recent observations in several laboratories that antibody titers to Borrelia burgdorferi are also elevated in several other skin diseases and that the spirochete can be detected in tissue sections of different organs may imply extension of the dermatological spectrum of Lyme disease . The significance of these findings in such heterogeneous diseases as morphea, lichen sclerosus et atrophicans, etc . however awaits final examination. Eur J Biochem, 1988 Mar 1, 172(2), 413 - 9 Amino acid sequence determination and three-dimensional modelling of thioredoxin from the photosynthetic bacterium Rhodobacter sphaeroides Y; Clement-Metral JD et al.; The complete primary structure of thioredoxin from Rhodobacter sphaeroides Y has been determined by analysis of peptides after cleavage with cyanogen bromide, chymotrypsin and trypsin . Peptides were separated by HPLC and analyzed by liquid-phase and gas-phase sequencer degradations . The protein consists of 105 residues (Mr = 11,180); its amino acid sequence shows a clear homology to the five known thioredoxins from plant or bacterial sources, with 40-56% residue identity when the proteins are aligned at the active-site disulfide . Not only the active-site regions are conserved, but also residues which belong to the hydrophobic surface suggested to be important for binding of procaryote thioredoxins in redox interactions with other proteins (residues 75-76; 91-93 in Escherichia coli) . A three-dimensional model of Rb . sphaeroides thioredoxin has been derived from the E . coli crystallographic structure with computer graphics . This model indicates that the overall structures as well as the active sites are closely similar; however, the residue substitutions allow both proteins to adopt different local folding as shown in the hydrophobic core. Acta Eur Fertil, 1988 Mar-Apr, 19(2), 93 - 7 The incidence of Chlamydia trachomatis-induced infection in women suffering from cervicovaginitis; Pungetti D et al.; The results of a research on Chlamydia T . (direct survey of both the antigen in the uterine cervix and plasmatic antibodies) in a group of subjects suffering for cervico-vaginitis are provided . The incidence of the Chlamydia infection (proved by either the presence of this bacterium or antibody positivity) is not different from the values reported in literature . Conversely, the presence of neither specific cytological or colposcopic patterns nor of priviledged comites at vaginal level could be demonstrated . Our data, however, confirm a greater incidence of this infection in women reporting early sexual life and a high number of partners . As for the relationship between Chlamydia and contraceptives a slightly higher incidence of positivity in the cervix of patients using oestro-progestinics was registered, whereas no significant difference was noted in the use of other contraceptives IUD includedPIP: Physicians examined 173 sexually active, non pregnant women suffering from lower genitalia inflammation . They responded to questions pertaining to their past and recent obstetric/gynecological history, to their partner's possible urogenital inflammations, the age of 1st intercourse, number of partners, and contraceptive use . 27.2% of the patients tested positive using immunoenzymatic techniques for Chlamydia trachomatis (CT) . No specific symptoms of CT were observed . A correlation exists between early sexual intercourse and a large number of partners and a greater incidence of CT infections . Almost 98% of all CT positive patients reported 1st sexual intercourse between 16 and 21 years . Antibody positivity ranged from 33% (1st intercourse before 15 years) to 24% (1st intercourse between 16-21 years) and decreased to 5.89% when 1st intercourse occurred 21 years . In addition, CT positive patients had many partners . A greater positivity in the cervix occurred in those using oral contraceptives, however . On the other hand, no positivity was noted for those who used IUDs . Those women who used several contraceptives, such as oral contraceptives, IUD, and barrier methods, had a higher incidence of CT positivity (53.2%) than other groups . Perhaps this is due to clinical cervicovaginitis symptoms prompting the women to change techniques . Specific colposcopy patterns and cytological alterations which some physicians believe indicate CT infections did not identify patients with Chlamydia . These data suggest that it is impossible to make a diagnosis based on symptoms, past sexual history, and contraceptive use . Therefore the data indicate that immunoenzymatic tests are needed . Can J Microbiol, 1988 Mar, 34(3), 287 - 98 The role of bacterial hydrophobicity in infection: bacterial adhesion and phagocytic ingestion; Absolom DR; The role that bacterial surface hydrophobicity (surface tension) plays in determining the extent of adhesion of polymer substrates and phagocytic ingestion is reviewed . The early attachment phase in bacterial adhesion is shown to depend critically on the relative surface tensions of the three interacting phases; i.e., bacteria, substrate, and suspending liquid surface tension . When suspended in a liquid with a high surface tension such as Hanks balanced salt solution, the most hydrophobic bacteria adhere to all surfaces to the greatest extent . When the liquid surface tension (gamma LV) is larger than the bacterial surface tension (gamma BV), then for any single bacterial species the extent of adhesion decreases with increasing substrate surface tension (gamma SV) . When gamma LV less than gamma BV then adhesion increases with increasing gamma SV . Bacterial surface tension also determines in part the extent of phagocytic ingestion and the degree to which antibodies specifically adsorb onto the bacterium resulting in opsonization . The nonspecific adsorption of antibodies results in a considerable modification in the surface properties of the bacteria . Bacterial surface hydrophobicity can be altered significantly through exposure to subinhibitory concentrations of antibiotics, surfactants, lectins, etc . The effect of these changes on subsequent phagocytic ingestion is discussed. Infect Immun, 1988 Mar, 56(3), 673 - 81 Appearance of sialoglycoproteins in encysting cells of Entamoeba histolytica; Chayen A et al.; Amoeba-bacterium cultures of Entamoeba histolytica transferred to a hypoosmotic medium depleted of nutrients changed morphologically and biochemically . The cells ejected grains of rice starch, rounded up, and formed a distinct cell wall that was resistant to detergent, bound the sialic acid-specific lectin from Limulus polyphemus, and became fluorescent with Calcofluor M2R . A subpopulation of these cells displayed more than one nucleus . All these signs are characteristic of encysting cells and were also observed in cysts obtained from a human patient . The morphological changes were accompanied by the appearance of two new glycoproteins with apparent molecular sizes of 100 and 150 kilodaltons which contained sialic acid . Sialic acid has been reported to be absent from trophozoites of Entamoeba species . The presence of this sugar residue on cyst-specific proteins parallels recently reported findings during the encystation of the related reptilian parasite Entamoeba invadens . This may indicate a basic role for sialic acid in the encystation of Entamoeba parasites. Cell, 1988 Feb 26, 52(4), 599 - 607 At least three different RNA polymerase holoenzymes direct transcription of the agarase gene (dagA) of Streptomyces coelicolor A3(2); Buttner MJ et al.; Using a combination of gel filtration and anion exchange FPLC, three different RNA polymerase holoenzymes from Streptomyces coelicolor A3(2) have been separated . Each holoenzyme transcribes from only one of the four promoters of the S . coelicolor A3(2) dagA gene . Holoenzyme reconstitution experiments identified the sigma factors responsible for recognition of two of the promoters . The previously identified E sigma 49 transcribes from the dagA p3 promoter, whereas a novel species, E sigma 28, recognizes the dagA p2 promoter . Circumstantial evidence suggests that the third holoenzyme, which transcribes from the dagA p4 promoter, is the previously identified E sigma 35 . This level of transcriptional complexity supports the idea that RNA polymerase heterogeneity may play a central role in the regulation and coordination of gene expression in this biochemically and morphologically complex bacterium. J Immunol, 1988 Feb 15, 140(4), 1022 - 7 IFN-gamma regulates the isotypes of Ig secreted during in vivo humoral immune responses; Finkelman FD et al.; The lymphokine IFN-gamma has been shown in vitro to stimulate IgG2a secretion and inhibit IgG1 and IgE secretion by LPS-activated B lymphocytes . To determine whether IFN-gamma has a similar isotype regulatory role in vivo, we studied the abilities of rIFN-gamma and a mAb to IFN-gamma to modify the isotypes of Ig secreted in mice injected with a goat antibody to mouse IgD, which by itself induces large increases in levels of serum IgG1 and IgE and a relatively small increase in serum IgG2a . Multiple injections of IFN-gamma substantially inhibited production of IgG1 and IgE, and stimulated production of IgG2a in affinity purified goat antibody specific for mouse IgD-treated mice; anti-IFN-gamma antibody blocked the effects of IFN-gamma and in fact enhanced IgG1 and IgE secretion and inhibited the IgG2a response in these mice . The role of IFN-gamma in the selection of isotypes of Ig produced in response to injection of mice with the bacterium Brucella abortus (BA) was also studied, because killed, fixed BA are known to stimulate IFN secretion and a predominantly IgG2a antibody response . Anti-IFN-gamma antibody strongly suppressed IgG2a secretion and stimulated IgG1, but not IgE, secretion in BA-immunized mice . BA suppressed IgG1 and IgE secretion and enhanced IgG2a secretion in affinity purified goat antibody specific for mouse IgD-injected mice; treatment of these mice with anti-IFN-gamma antibody reversed the effects of BA on IgG1 and IgG2a secretion, but not the suppressive effect of BA on IgE secretion . These observations demonstrate that IFN-gamma has an important and perhaps unique physiologic role in the stimulation of IgG2a secretion and in the suppression of secretion of IgG1, whereas bacterial antigens can suppress IgE secretion by other mechanisms in addition to IFN-gamma secretion. Infect Immun, 1988 Feb, 56(2), 395 - 9 Effect of dietary restriction on total and bacterium-specific mucosal secretory immunoglobulin A in bile-diverted intestinal self-filling blind loops; Lichtman SN et al.; The effect of starvation on the mucosal secretory immunoglobulin A (sIgA) response to bacterial antigens was studied in bile-free rat self-filling blind loops constructed at the end of a Roux-en-Y branch of jejunum . Rats were fed a 50% restricted diet for 1 to 4 weeks after surgery . sIgA was measured in the mucosa and lumen by an enzyme-linked immunosorbent assay . Dietary restriction caused a final rise of luminal sIgA which was less than 50% of that of normally fed controls Luminal bacteria counts were not different in the two groups . The percentage of total sIgA precipitated with intestinal bacteria was not significantly affected by dietary restriction, and there was no change in the specific binding of sIgA to several bacterial species . Nonprecipitated sIgA exhibited a low but significant specific binding to bacteria in both diet-restricted and fed rats . Diet restriction therefore reduced the total sIgA response to luminal bacteria, but the specific bacterial binding capacity per microgram of sIgA was not altered . In these short-term experiments diet-restricted animals appeared to be capable of secreting sIgA in excess of requirement, since the nonprecipitable luminal fraction contained free sIgA with binding capacity for bacteria . The ability of sIgA to react with specific antigens may therefore be of more significance as an indicator of bacterial susceptibility than the measurement of total sIgA. J Gen Virol, 1988 Feb, 69 ( Pt 2), 385 - 93 Transcriptional mapping of the bacteriophage Mu DNA; Barron C et al.; The transcription of temperate phage Mu throughout lytic development was analysed quantitatively by hybridization of pulse-labelled RNA to full-length Mu DNA and to plasmids that define Mu DNA segments covering the whole phage genome . The transcription rate (i.e . binding data corrected for the incorporation rate of the radioactive precursor, for the size of the DNA template, and for the number of phage genomes present in the bacterium at the time of analysis) revealed three defined phases of Mu transcription: early (0 to 9 min), intermediate (between 9 and the interval 14 to 17 min) and late (from the interval 14 to 17 min onward) . The analysis also revealed that the region comprising the genes involved in phage morphogenesis was organized into two independent 'late' transcription units. J Bioenerg Biomembr, 1988 Feb, 20(1), 41 - 58 Molecular genetics of F1-ATPase from Escherichia coli; Futai M et al.; We have reviewed recent molecular biological studies on F1-ATPase of Escherichia coli and emphasized the advantages of using the bacterium in studies on this important enzyme . All subunits had homologies of varied degrees with those from other organisms . Mutations of F1 subunits caused defects in catalysis and assembly . Defects of the mutant enzymes were studied extensively together with the determination of the amino acid substitutions . Extensive molecular biological studies may help greatly in understanding the normal mechanism and assembly of the complex. Histochem J, 1988 Feb, 20(2), 108 - 16 Pitfalls in ecto-5'-nucleotidase enzyme cytochemistry as demonstrated by the immunogold-labelling technique on macrophages; Cramer EM et al.; We have used both the enzyme cytochemical method with lead nitrate as a capture agent and an immunological method at the electron microscope level to localize plasma membrane 5'-nucleotidase in rat peritoneal resident macrophages during the initial interactions of latex beads or heat-killed Escherichia coli with the cell during phagocytosis . In macrophages at rest, cytochemical reaction product was evenly distributed along the external surface of the plasma membrane . However, when the cells were phagocytosing latex beads or bacteria, reaction product covered the entire surface of the adhering particles . To determine whether the apparent redistribution of 5'-nucleotidase onto the adhering particle was fact or artifact, we localized 5'-nucleotidase using a monoclonal antibody and an immunogold labelling technique . In macrophages binding or beginning to ingest bacteria, gold particles were distributed along the plasma membrane, except at the sites of cell-bacterium internalization . More significantly, the adhering bacteria were free of gold particles and therefore had no 5'-nucleotidase on their surfaces . Latex beads proved to be unsuitable as a test particle because the gold particles stuck to them non-specifically . We conclude that the artifactual redistribution of lead-phosphate reaction product is a major drawback of enzyme cytochemical methods when used on cell surfaces and that the immunogold labelling technique is more reliable. Eur J Biochem, 1988 Jan 15, 171(1-2), 67 - 72 Purification and characterization of a bacterial dehalogenase with activity toward halogenated alkanes, alcohols and ethers; Janssen DB et al.; An enzyme that is capable of hydrolytic conversion of halogenated aliphatic hydrocarbons to their corresponding alcohols was purified from a 1,6-dichlorohexane-degrading bacterium . The dehalogenase was found to be a monomeric protein of relative molecular mass 28,000 . The affinity for its substrates was relatively low with Km values for short-chain haloalkanes in the range 0.1-0.9 mM . The aliphatic dehalogenase showed a much broader substrate range than has been reported for halidohydrolases so far . Novel classes of substrates include dihalomethanes, C5-C9 1-halo-n-alkanes, secondary alkylhalides, halogenated alcohols and chlorinated ethers . Several of these compounds are important environmental pollutants, e.g . methylbromide, dibromomethane, 1,2-dibromoethane, 1,3-dichloropropene, and bis(2-chloroethyl)ether . The degradation of chiral 2-bromoalkanes appeared to proceed without stereochemical preference . Optically active 2-bromobutane was converted with inversion of configuration at the chiral carbon atom, suggesting that the dehalogenase reaction proceeds by a nucleophilic substitution involving a carboxyl group or base catalysis. Eur J Biochem, 1988 Jan 15, 171(1-2), 245 - 52 The hopanoids of the purple non-sulfur bacteria Rhodopseudomonas palustris and Rhodopseudomonas acidophila and the absolute configuration of bacteriohopanetetrol; Neunlist S et al.; Five complex hopanoids have been detected in the purple non-sulfur bacterium Rhodopseudomonas acidophila . Next to the polyfunctionalized methylcyclopentane bacteriohopanetetrol ether already isolated from Methylobacterium organophilum, 35-carbamoylbacteriohopane-32,33,34-triol, 34,35-dicarbamoylbacteriohopane-32,33-diol and two nucleoside analogues, (22R)-30-(5'-adenosyl)hopane and (22S)-30-(5'-adenosyl)hopane were isolated and identified by spectroscopic and chemical methods . In Rhodopseudomonas palustris, however, only 35-amino-bacteriohopane-32,33,34-triol was detected . Chemical correlation between adenosylhopane and bacteriohopanetetrol, as well as comparison of derivatives obtained from bacterial and synthetic hopanoids, permitted the determination of the configurations of all asymmetric centres of the side-chain of bacteriohopanetetrol as 22R, 32R, 33R and 34S . According to the stereochemistry, this side-chain could be a D-ribose derivative linked through its C-5 carbon atom to the hopane skeleton. Eur J Biochem, 1988 Jan 15, 171(1-2), 351 - 4 Engineered rat insulin I analogue having a B16 Tyr/Asp replacement exhibits unchanged susceptibility to cleavage by insulin proteinase; Varley JM et al.; An analogue of rat insulin I was produced by oligonucleotide-directed mutagenesis of a cloned rat preproinsulin I cDNA, followed by expression of a resulting mutant gene in Escherichia coli K-12 and proteolytic cleavage of mutant proinsulin isolated from this bacterium . The Tyr-to-Asp replacement at residue B16 in the insulin analogue had been expected to diminish the rate of cleavage of the molecule by the enzyme insulin proteinase, since the bond TyrB16-LeuB17, invariant in all mammalian species, had been proposed by other authors as one of the early, major sites of proteolytic attack . In the event the substitution had no measurable effect on the rate of degradation by insulin proteinase . Thus we find no support in these experiments for the hypothesis that the site in question is of primary importance in the degradation of rat insulin I by the enzyme. Br J Rheumatol, 1988, 27 Suppl 2, 32 - 3 Cross-reactivity studies on bacteria believed to be associated with inflammatory bowel disease (IBD), ankylosing spondylitis (AS) and reactive arthritis (ReA); Pease PE et al.; Six bacteria believed to be associated with IBD and/or AS and ReA were investigated for antigenic cross-reactivity using an ELISA . Considerable degrees of cross-reactivity were detected and it is suggested that if bacterial antigens are involved aetiologically in the diseases, no specific bacterium is concerned. J Biochem (Tokyo), 1988 Jan, 103(1), 121 - 5 Purification and characterization of ferredoxin from Desulfovibrio vulgaris Miyazaki; Ogata M et al.; Two ferredoxins, Fd I and Fd II, were isolated and purified from Desulfovibrio vulgaris Miyazaki . The major component, Fd I, is an iron-sulfur protein of Mr 12,000, composed of two identical subunits . The absorption spectra of Fd I and Fd II have a broad absorption shoulder near 400 nm characteristic of iron-sulfur proteins . The purity index, A400/A280, of Fd I is 0.69, and its millimolar absorption coefficient at 400 nm is 3.73 per Fe . It contains two redox centers with discrete redox behaviors . The amino acid composition and the N-terminal sequence of Fd I are similar to those of Fd III of Desulfovibrio africanus Benghazi and Fd II of Desulfovibrio desulfuricans Norway . Fd I does not serve as an electron carrier for the hydrogenase of D . vulgaris Miyazaki, but it serves as a carrier for pyruvate dehydrogenase of this bacterium . The evolution of H2 from pyruvate was observed by a reconstructed system containing purified hydrogenase, cytochrome C3, Fd I, partially purified pyruvate dehydrogenase, and CoA . The H2-sulfite reducing system can be reconstructed from the purified hydrogenase, cytochrome C3, Fd I and desulfoviridin (sulfite reductase), but the reaction rate is very slow compared to that of the crude extract at the same molar ratio of the components. J Wildl Dis, 1988 Jan, 24(1), 176 - 7 Actinobacillosis in free-ranging snowshoe hares (Lepus americanus) from Alaska; Zarnke RL et al.; Actinobacillus capsulatus was isolated from lung, liver, and/or spleen tissue of three snowshoe hares (Lepus americanus) in Alaska . This is the first report of the isolation of this bacterium from free-ranging hares . Actinobacillus capsulatus may have a negative impact on the population density of hares. Appl Environ Microbiol, 1988 Jan, 54(1), 153 - 7 Existence of a new type of sulfite oxidase which utilizes ferric ions as an electron acceptor in Thiobacillus ferrooxidans; Sugio T et al.; A new type of sulfite oxidase which utilizes ferric ion (Fe3+) as an electron acceptor was found in iron-grown Thiobacillus ferrooxidans . It was localized in the plasma membrane of the bacterium and had a pH optimum at 6.0 . Under aerobic conditions, 1 mol of sulfite was oxidized by the enzyme to produce 1 mol of sulfate . Under anaerobic conditions in the presence of Fe3+, sulfite was oxidized by the enzyme as rapidly as it was under aerobic conditions . In the presence of o-phenanthroline or a chelator for Fe2+, the production of Fe2+ was observed during sulfite oxidation by this enzyme under not only anaerobic conditions but also aerobic conditions . No Fe2+ production was observed in the absence of o-phenanthroline, suggesting that the Fe2+ produced was rapidly reoxidized by molecular oxygen . Neither cytochrome c nor ferricyanide, both of which are electron acceptors for other sulfite oxidases, served as an electron acceptor for the sulfite oxidase of T . ferrooxidans . The enzyme was strongly inhibited by chelating agents for Fe3+ . The physiological role of sulfite oxidase in sulfur oxidation of T . ferrooxidans is discussed. Biomed Biochim Acta, 1988, 47(6), 537 - 42 Polyclonal IgG synthesis of mononuclear cells induced by the homopolymer of 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl units isolated from Brucella melitensis; Montag T et al.; Brucella melitensis attaches selectively to B lymphocytes . The bacterium induces in vitro only a slight proliferation of mononuclear cells and inhibits the spontaneous IgG synthesis . In order to characterize these effects, the O-chain of the lipopolysaccharide of Brucella melitensis (a polysaccharide consisting of poly-4,6-dideoxy-4-formamido-alpha-D-mannoside) was prepared and its influence on lymphocyte functions tested . This compound had no effect on blast transformation whereas IgG synthesis of the cells was stimulated more than tenfold by the addition of the Brucella polymannoside. Proteins, 1988, 3(3), 184 - 6 Crystallization and preliminary X-ray diffraction study of a protein with a high potential rubredoxin center and a hemerythrin-type Fe center; Sieker LC et al.; A newly discovered iron-containing protein, isolated from the bacterium Desulfovibrio vulgaris (Hildenborough, NCIB 8303), has been crystallized . The molecule appears to be a dimer of mass 44kDa . This protein has iron centers with spectrascopic similarities to those in rubredoxins and in hemerythrins . The X-ray diffraction shows symmetry consistent with space group I222 or I212121 . Cell parameters are a = 49.2 A, b = 81.3 A, c = 100.1 A, and alpha, beta, gamma = 90 degrees . X-ray diffraction data have been collected to 3.0 A, and a search for useful heavy atom derivatives is in progress for the analysis of the crystal structure of this Fe-protein. Antonie Van Leeuwenhoek, 1988, 54(6), 483 - 96 Bilophococcus magnetotacticus gen . nov . sp . nov., a motile, magnetic coccus; Moench TT; The morphological, biochemical, and magnetotactic properties of a single magnetic bacterium are reported . Although this bacterium has not been cultured axenically, the unusual magnetotactic behavior has allowed the collection of cell material of sufficient quantity and purity to allow characterization . The results indicate that this organism represents a new genus of colorless, sulfur-depositing bacteria, albeit of uncertain affiliation . The name proposed for this new genus/species, Bilophococcus magnetotacticus, reflects the most distinctive traits of morphology, motility, and magnetic mineral formation . Classification is based on type descriptive material. Adv Virus Res, 1988, 35, 219 - 49 Viral-bacterial synergistic interaction in respiratory disease; Babiuk LA et al.; In the present review we have identified how viruses can alter the host's susceptibility to bacterial infections by altering both environmental conditions in the lung which favor bacterial replication as well as by suppressing the host's defense mechanisms which prevent clearance of the bacteria . In many instances, these interactions are extremely complex but similar for many viruses . If the virus can overcome the initial host defense mechanisms, which include local antibody and mucus, the virus initiates tissue damage as a result of direct replication within the epithelial cells lining the mucosal surfaces of the respiratory tract . As a result of virus infection, the host cells respond by producing a variety of mediators including various types of interferons, which can alter both virus replication and host response . Replication also produces by-products of virus infection capable of initiating an inflammatory process, which in turn, through release of other mediators, can further modify lung defense mechanisms and encourage bacterial adherence and growth . The bacterium, in turn, releases chemotactic factors which encourage infiltration of specific effector cells into the lung . These effector cells can cause tissue damage and immunopathology, which encourage rapid bacterial growth and may result in death of the animal . In order to be able to control this complicated scenario, it is important either to prevent the initial infection with viruses or to reduce the degree of immunosuppression, so that bacterial clearance can occur rapidly before microcolony formation and extensive lung damage occur . Once a large amount of bacterial replication and lung damage is present, the use of antibiotics is generally of limited value . A schematic illustration of the complexity of the various interactions and counteractions occurring during virus--bacterial synergistic interactions is presented in Fig . 1. Bull Soc Pathol Exot Filiales, 1988, 81(4), 726 - 31 {A new sensitivity test for substances used as antiseptics or disinfectants . I . Biological studies}; Dodin A et al.; In this first article, the authors present a fast method of determining the bactericide and bacteriostatic power of the germs, based on the disparition of the succino-dehydrogenase activity of a bacterium, objectified by a neotetrazolium chloride reaction . This reaction allows a colorimetric qualitative and quantitative determination of the activity of a preparation and the duration of its contact with the bacteria . This method allows a study in electronic microscopy which will be published in a second article. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1988 Jan, 185(4-5), 350 - 67 {The use of UV rays for the disinfection of water . I . Microbiologic studies of drinking water}; Martiny H et al.; As a physical disinfection method without chemicals required the ultraviolet irradiation was tested for disinfection of drinking water . The survival was measured as a function of exposure to radiation for S . enteritidis, E . cloacae, C . freundii, S . marcescens, E . coli, K . pneumoniae und S . faecium . The bacteria were grown in trypton soya broth until they were well into the exponential phase . Two different UV-disinfection units were tested . Both consist of cylindrical shaped chambers with one low-pressure mercury-discharge lamp with their longitudinal axis parallel to the chambers . With 10(6) cfu/ml the experiments were done with three different rates of flow of 7,2 m3/h, 4,0 m3/h and 2,0 m3/h . The minimum exposures to radiation necessary to cause a 99.999% reduction were 10-86 mWs/cm2 depending on the test bacterium and on the UV-disinfection unit . The minimum doses ranged for S . enteritidis up to 13 mWs/cm2, for E . coli up to 21 mWs/cm2, for K . pneumoniae up to 39 mWs/cm2, for S . faecium up to 42 mWs/cm2, for E . cloacae up to 43 mWs/cm2, for C . freundii up to 72 mWs/cm2, and for S . marcescens up to 86 mWs/cm2. Arch Biochem Biophys, 1988 Jan, 260(1), 134 - 8 Lysine and arginine transport in the photosynthetic bacterium Chromatium vinosum; Kim YA et al.; The photosynthetic purple sulfur bacterium Chromatium vinosum can take up both arginine and lysine in the light and, to a lesser extent, in the dark . Competitive inhibition experiments suggest the likely presence of two transport systems in this bacterium: One capable of transporting either lysine or arginine and a second capable of transporting arginine but not lysine . Uptake of both amino acids is electrogenic and appears to involve the cotransport of neither protons nor sodium ions . It is suggested that the transport occurs via an electrogenic uniport. Cold Spring Harb Symp Quant Biol, 1988, 53 Pt 1, 11 - 7 Roles of methylation and phosphorylation in the bacterial sensing system; Koshland DE Jr et al.; Chemotaxis is an intriguing model system for the study of second-messenger pathways . One of the puzzles of second-messenger pathways in eukaryotic cells has been that many of these pathways interact, with one pathway either desensitizing or sensitizing an alternate messenger pathway . The chemotaxis system offered a particularly interesting chance to analyze such systems, because one of them is a methylation pathway and the other a phosphorylation pathway . The above description indicates that these two pathways interact with each other in a highly sophisticated way to produce an extremely important survival system . The stimulus on a receptor activates an excitation system that operates through phosphorylation, or the inhibition of phosphorylation, depending on whether the stimulus is a repellent or an attractant . That system ultimately generates or inhibits phosphorylation of a small peptide, the CheY protein, which apparently is the response regulator . The instant that this fast excitation is generated by a change in gradient, the change in conformation of the protein sets in motion a second process, i.e., the adaptation . That is a device which, over a longer period of time, has two functions: It serves as the comparator, which allows the comparison of the past with the present, essential for deletion of a gradient; it also sets in motion the reset to zero, so that the bacterium will not be overwhelmed by any one stimulus but can use all of its receptors to optimize its environment . These two systems by themselves are adequate for chemotaxis, but there is a further elegent complexity: The excitation system feeds back into the adaptation system to produce an asymmetry in the responses . The reason for that asymmetry is that the bacterium wishes to travel in the wrong direction only long enough to produce a detectable signal that it is migrating incorrectly . It wishes to keep swimming in the positive direction as long as the signals indicate that the direction is favorable . Hence, the feedback between the phosphorylation and methylation system involves further fine tuning. Acta Otolaryngol Suppl, 1988, 454, 167 - 74 Bacterial adherence and upper respiratory tract disease: a correlation between S . pyogenes attachment and recurrent throat infections; Galioto GB et al.; The attachment of bacteria to mucosal surfaces is the initial event in the pathogenesis of infectious diseases . The authors present a study about the adherence of strain of S . pyogenes isolated in subjects with recurrent tonsillitis . A correlation was found between the adherence ability of the bacterium and the number of episodes per year . The study about the SIgA, the infection development and the bacterium adherence showed a direct correlation between SIgA levels and the number of phlogistic episodes . The importance of the role assumed by bacterial adherence in the genesis of the phlogistic process is to be emphasized yet again. Adv Exp Med Biol, 1988, 239, 1 - 11 Lymphokine regulation of macrophage effector activities; Nacy CA et al.; Our concept of the regulation of macrophage activation is ever expanding and contracting . In regard to the number of LK that regulate macrophages killing activities, we have entered a new phase . In the beginning there was one macrophage activation factor, MIF; then there were many macrophage activation factors, most uncharacterized and bearing a variety of names . Then came IFN, a genetically cloned single reagent that induced destruction of virtually every target assessed; all activities of macrophages were assumed to be regulated by IFN . Once again, however, the LK universe is expanding: the number of single, cloned reagents that induce macrophage killing activities is amazing . With just two targets, a fibrosarcoma cell and an intracellular amastigote of L . major, we can identify 5 different macrophage activation factors, four of which are cloned and sequenced . As more recombinant reagents become available, the story of macrophage activation is likely to become even more complex . It is fascinating not only that certain of the LK are capable of inducing single effector reactions in the absence of effects on other effector activities, but also that at least one effector reaction requires the cooperation of several molecularly distinct LK . The complexity of LK activation factors that regulate a single effector reaction in vitro is compounded by the complexity in effector cell populations . For example, inflammatory macrophages exposed to LK kill the fibrosarcoma tumor target 5 to 10-fold better than an equal number of resident peritoneal macrophages . In contrast, LK treated resident macrophages eliminate intracellular amastigotes of leishmania far more efficiently than inflammatory cells . Thus, changes in cell populations dramatically affect the capacity to demonstrate a single effector reaction . Further, simple changes in assay conditions also determine whether an effector reaction can be observed in vitro . And superimposed upon all these layers of complexity is the target itself . The mechanisms a macrophages uses to block the replication of a virus may be totally ineffective in the destruction of a multicellular helminth, such as Schistosoma mansoni . And there is no reason to suspect that the extracellular destruction of a tumor target occurs by the same means that the macrophage uses to kill an intracytoplasmic bacterium, such as a rickettsia.(ABSTRACT TRUNCATED AT 400 WORDS) Cell Motil Cytoskeleton, 1988, 11(1), 46 - 63 Characterization of gliding motility in Flexibacter polymorphus; Ridgway HF et al.; Motility of the marine gliding bacterium Flexibacter polymorphus was studied by using microcinematographic techniques . Following adhesion to a glass surface, multicellular filaments and individual cells usually began to glide within a few seconds at a speed of approximately 12 micron per second (at 23 degrees C) . Adhesion to the glass surface was evidently mediated by multitudes of extremely fine extracellular fibrils . Gliding velocity was independent of filament length but directly related to electron-transport activity and substratum temperature in the range 3-35 degrees C . The rate of gliding was inversely related to medium viscosity, suggesting that the locomotor apparatus functions at constant torque . Forward motion was occasionally interrupted by direction reversals, somersaults (observed primarily in single cells of short filaments), or spinning of filaments tethered by one pole . The frequency of direction reversal was found to be an inverse function of filament length . Translational motility was invariably accompanied by sinistral revolution about the longitudinal axis of a filament . The sense and pitch of revolution were constant among filaments of different length . Polystyrene microspheres or India ink particles adsorbed to gliding cells were actively displaced in either direction, their movement tracing either a regular zigzag or helical path along the filament surface . Because microspheres were also observed to move on nonmotile filaments, particle translocation was evidently not obligatorily linked to gliding locomotion . Multiple particles adsorbed to a single filament often moved independently . The data are consistent with a motility mechanism involving limited motion in numerous mechanically independent (yet functionally coordinated) domains on the cell surface. Proteins, 1988, 4(1), 63 - 70 Model of a complex between the tetrahemic cytochrome c3 and the ferredoxin I from Desulfovibrio desulfuricans (Norway strain); Cambillau C et al.; A three-dimensional model of an electron-transfer complex between the tetrahemic cytochrome c3 and the ferredoxin I from the sulfate-reducing bacterium Desulfovibrio desulfuricans (Norway strain) has been generated through computer graphics methods . The model is based on the known X-ray structure of the cytochrome and on a model of the ferredoxin that has been derived through computer graphics modeling and energy minimization methods, from the X-ray structure of the homologous ferredoxin from Peptococcus aerogenes . Four possible models of interaction between the two molecules were examined by bringing in close proximity each of the four hemes and the redox center (4Fe-4S) of the ferredoxin and by optimizing the ion pairs interactions . One of these models shows by far the "best" structure in terms of charges, interactions, and complementarity of the topology of the contact surfaces . In this complex, the distance between the iron atoms of the ferredoxin redox center and the hemic iron atom is 11.8 A, which compares well with those found between redox centers in other complexes . The contact surface area between the two molecules is 170 A2. Virchows Arch A Pathol Anat Histopathol, 1988, 412(6), 563 - 72 Cat scratch disease . An epidemiological and ultrastructural study of lymphadenitis caused by Warthin-Starry positive bacteria; Kudo E et al.; The aetiological agent of cat scratch disease (CSD) has been unknown for more than 30 years . Recently, a micro-organism clearly shown with Warthin-Starry silver (W-S) stain was found and thought to be a possible cause of the disease . In this study, 32 cases of regional lymphadenopathy histologically compatible with CSD and 20 contrasting cases of lymphadenopathy were examined retrospectively with W-S stain . W-S positive pleomorphic organisms were clearly demonstrated in 20 of the 32 suspected cases of CSD, but in none of the other cases . The onset of disease in these 20 cases with W-S positive organisms occurred between July and January . This seasonal variation in the onset of disease was highly significant (P less than 0.005) and was not due to a single epidemic . Moreover, some characteristic morphological features of the organism were found by electron microscopic observations . Ultrastructurally, the organism was a bacterium showing a chain-like arrangement, septal formation, branching and clubbed ends. J Immunol, 1987 Dec 15, 139(12), 4203 - 7 Lysis of cells infected with typhus group rickettsiae by a human cytotoxic T cell clone; Carl M et al.; Cytolytic human T cell clones generated in response to the intracellular bacterium Rickettsia typhi were characterized . Growing clones were tested for their ability to proliferate specifically in response to antigens derived from typhus group rickettsiae or to lyse targets infected with R . typhi or Rickettsia prowazekii . Two clones were able to lyse targets infected with typhus group rickettsiae . One of these clones was more fully characterized because of its rapid growth characteristics . This cytolytic clone was capable of lysing an autologous infected target as well as a target matched for class I and II histocompatibility leukocyte antigens (HLA) . It was not capable, however, of lysing either a target mismatched for both class I and II HLA or a target partially matched for class I HLA . In addition, the clone exhibited specificity in that it was able to lyse an autologous target infected with typhus group rickettsiae, but did not lyse an autologous target infected with an antigenically distinct rickettsial species, Rickettsia tsutsugamushi . These results demonstrate, for the first time, that cells infected with intracellular bacteria can be lysed by human cytotoxic T lymphocytes. Biochim Biophys Acta, 1987 Dec 14, 922(3), 287 - 93 Inhibition of acetoacetyl-CoA synthetase from rat liver by fatty acyl-CoAs; Ito M et al.; The activity of acetoacetyl-CoA synthetase from rat liver was found to be negatively regulated by coenzyme A, fatty acyl-CoAs and acetoacetyl-CoA in vitro . With increasing concentrations of coenzyme A (substrate inhibition occurring at concentrations higher than 50 microM) the pH optimum shifted toward the acidic side (7.5-8.5 with 5 microM coenzyme A and 6.5-7.0 with 500 microM coenzyme A), in parallel with progressively decreasing enzyme activity . Fatty acyl-CoAs of various chain lengths dose-dependently inhibited acetoacetyl-CoA synthetase from rat liver, but much less effectively a similar enzyme from a bacterium, Zoogloea ramigera I-16-M . Palmitoyl-CoA, the most potent inhibitor of the rat liver enzyme, with an apparent Ki value of 9.8 microM, apparently inhibited the enzyme below its critical micellar concentration, not due to its detergent action . Acetoacetyl-CoA showed product inhibition with a Ki value of 15 microM . These results suggest a possible physiological regulation mechanism for this enzyme with respect to fatty acid biosynthesis. J Bacteriol, 1987 Dec, 169(12), 5801 - 7 Involvement of transport in Rhodobacter sphaeroides chemotaxis; Ingham CJ et al.; The chemotactic response to a range of chemicals was investigated in the photosynthetic bacterium Rhodobacter sphaeroides, an organism known to lack conventional methyl-accepting sensory transduction proteins . Strong attractants included monocarboxylic acids and monovalent cations . Results suggest that the chemotactic response required the uptake of the chemoeffector, but not its metabolism . If a chemoeffector could block the uptake of another attractant, it also inhibited chemotaxis to that attractant . Sodium benzoate was not an attractant but was a competitive inhibitor of the propionate uptake system . Binding in an active uptake system was therefore insufficient to cause a chemotactic response . At different concentrations, benzoate either blocked propionate chemotaxis or reduced the sensitivity of propionate chemotaxis, an effect consistent with its role as a competitive inhibitor of uptake . Bacteria only showed chemotaxis to ammonium when grown under ammonia-limited conditions, which derepressed the ammonium transport system . Both chemotaxis and uptake were sensitive to the proton ionophore carbonyl cyanide m-chlorophenylhydrazone, suggesting an involvement of the proton motive force in chemotaxis, at least at the level of transport . There was no evidence for internal pH as a sensory signal . These results suggest a requirement for the uptake of attractants in chemotactic sensing in R . sphaeroides. Acta Pathol Jpn, 1987 Dec, 37(12), 1973 - 7 Nonspecific simple eosinophilic granulomatous prostatitis with eosinophilia in peripheral blood . A case report; Sugiura H et al.; Nonspecific simple eosinophilic granulomatous prostatitis is extremely rare and in the present paper, the first case showing eosinophilia in the peripheral blood is reported . The patient was a 55-year-old Japanese man who was admitted because of difficulty in urination over a period of several years . The laboratory findings revealed marked eosinophilia (36%) in the peripheral blood, but the patient's past history showed neither bronchial asthma nor any allergic tendency . Transurethral resection of the prostate was performed and the histopathologic findings revealed a picture of non-caseating granulomatous prostatitis with massive eosinophilic infiltration without fibrinoid necrosis or vasculitis . Fragments of the prostatic urethra also showed the same findings . No fungus, bacterium or parasite was found . Although remnants of smooth muscle fibers were noted in the granulomas, neither immunoglobulins nor complement components could be demonstrated, and the etiology remained undetermined. J Bacteriol, 1987 Dec, 169(12), 5808 - 14 Methylation-independent and methylation-dependent chemotaxis in Rhodobacter sphaeroides and Rhodospirillum rubrum; Sockett RE et al.; In vivo and in vitro methylation, methanol production assays, and the use of specific antibodies raised against the sensory transducing protein Tar in Escherichia coli all failed to demonstrate the presence of methyl-accepting chemotaxis proteins (MCPs) in the photosynthetic bacterium Rhodobacter sphaeroides, although such proteins did exist in another photosynthetic bacterium, Rhodospirillum rubrum . The range of chemicals to which Rhodobacter sphaeroides responds, the lack of an all-or-none response, and the lack of true repellents indicate an alternative chemosensory pathway . The existence of MCPs in Rhodospirillum rubrum means that the lack of MCPs is not the result of a phototrophic metabolism, but may be connected to the unidirectional flagellar motor of Rhodobacter sphaeroides. J Bacteriol, 1987 Dec, 169(12), 5575 - 8 Actinomycin synthesis in Streptomyces antibioticus: enzymatic conversion of 3-hydroxyanthranilic acid to 4-methyl-3-hydroxyanthranilic acid; Jones GH; A methyltransferase which utilizes 3-hydroxyanthranilic acid (HAA) as a substrate was identified in detergent-treated extracts of the bacterium Streptomyces antibioticus . The enzyme catalyzes the transfer of methyl groups from {14C}S-adenosylmethionine to HAA, but does not catalyze the methylation of 3-hydroxy-DL-kynurenine . Enzyme, substrate, time, and pH dependencies for the methyl transfer reaction were examined . Reaction products obtained from scaled-up reaction mixtures were fractionated by chromatography on Dowex 1, and the Dowex 1 fractions were examined by paper and thin-layer chromatography . One Dowex fraction was shown to contain a radioactive product with the chromatographic properties of 4-methyl-3-hydroxyanthranilic acid (MHA), a known intermediate in the biosynthesis of actinomycin . Available evidence indicates that the conversion of HAA to MHA is an early step in the biosynthesis of actinomycin by S . antibioticus and other actinomycin-producing streptomycetes. Eur J Biochem, 1987 Nov 16, 169(1), 167 - 73 The occurrence of 2'-5' oligoadenylates in Escherichia coli; Trujillo MA et al.; The use of a highly specific radioimmunoassay and of HPLC permitted us to confirm the occurrence of 2'-5' oligoadenylates {p chi (A2'p5')nA} in several strains of Escherichia coli . Cellular concentrations of 2'-5' oligoadenylates ranged from 50 nM to 300 nM . The mixture of 2'-5' oligoadenylates consisted primarily of pppA2'p5'A, pA2'p5'A,A2'p5'Ap and A2'p5'A under normal conditions of growth . None of them activated RNase L . Infection of the bacteria with the single-stranded DNA phage M13 or induction of a heat-inducible, non-lytic mutant of phage lambda led to a significant increase in the total pool of 2'-5' oligoadenylates, paralleling the progressive inhibition of growth . Likewise, the inhibition of protein synthesis with chloramphenicol stimulated the accumulation of 2'-5' oligoadenylates . Furthermore, the inhibition of bacterial growth by either phage or by chloramphenicol brought about a change in the composition of the 2'-5' oligoadenylate pool; 5'-phosphorylated 2'-5' oligoadenylates accumulated and became the major components . The findings indicate a parallelism between the effects of viral infection on the synthesis of 2'-5' oligoadenylates in eukaryotes and similar effects subsequent to phage growth in the bacterium E . coli. J Biol Chem, 1987 Nov 5, 262(31), 14983 - 9 Respiratory enzymes of Thiobacillus ferrooxidans . A kinetic study of electron transfer between iron and rusticyanin in sulfate media; Blake RC 2nd et al.; Thiobacillus ferrooxidans is a chemolithotrophic bacterium capable of fulfilling all of its energy requirements from the oxidation of soluble ferrous sulfate . Rusticyanin is a soluble blue copper protein found in abundance in the periplasmic space of this bacterium . The one-electron transfer reaction between soluble iron and purified rusticyanin has been studied by stopped flow spectrophotometry in acidic solutions containing sulfate . Second order rate constants for the reduction of rusticyanin by Fe2+, FeHSO4+, and FeSO4(0) were 0.022, 0.73, and 2.30 M-1 s-1, respectively . The pseudo-first order rate constant for the reduction of rusticyanin exhibited substrate saturation when the concentration of the total ferrous ion was varied in solutions of limiting sulfate . This saturation behavior was quantitatively described using the values of the second order rate constants listed above and the distribution of the total ferrous ion into its water-, bisulfate-, and sulfate-coordinated forms . Second order rate constants for the oxidation of rusticyanin by Fe3+ and FeSO4+ were 0.73 and 0.26 M-1 s-1, respectively . The electron transfer reactions between iron and rusticyanin monitored in vitro were far too slow to support the hypothesis that rusticyanin is the primary oxidant of ferrous ions in the iron-dependent respiratory electron transport chain of T . ferrooxidans. J Bacteriol, 1987 Nov, 169(11), 5279 - 88 Protein inclusions produced by the entomopathogenic bacterium Xenorhabdus nematophilus subsp . nematophilus; Couche GA et al.; The entomopathogenic bacterium Xenorhabdus nematophilus subsp . nematophilus produces two types of intracellular inclusion bodies during in vitro culture . Large cigar-shaped inclusions (designated type 1) and smaller ovoid inclusions (designated type 2) were purified from cell lysates, using differential centrifugation in discontinuous glycerol gradients and isopycnic density gradient centrifugation in sodium diatrizoate . The inclusions, composed almost exclusively of protein, are readily soluble at high and low pH values and in the presence of cation chelators such as EDTA, anionic detergents (sodium dodecyl sulfate), or protein denaturants (urea, NaBr) . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified inclusions revealed a single 26-kilodalton protein (IP-1) in type 1 inclusions and a 22-kilodalton protein (IP-2) in type 2 inclusions . Analysis of these proteins by isoelectric focusing in the presence of 8 M urea showed that IP-1 is acidic and IP-2 is neutral . Furthermore, each protein occurred in multiple forms differing slightly in isoelectric point . Other variations in peptides released by trypsin digestion, immunological properties, and amino acid composition revealed significant structural differences between IP-1 and IP-2 . Kinetic studies using light microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting procedures showed that inclusion protein synthesis occurs only during the second half of exponential culture growth . Synthesis of inclusion proteins and their aggregation to form inclusions occurred concurrently . Possible functions for these abundant proteins are discussed. Infect Immun, 1987 Nov, 55(11), 2701 - 5 Influence of growth temperature on virulence of Legionella pneumophila; Edelstein PH et al.; The effect of growth temperature on the virulence of a strain of broth-grown serogroup 1 Legionella pneumophila (Wadsworth F889) was examined by growing the bacterium at different temperatures and then infecting guinea pigs (by intratracheal injection) and guinea pig alveolar macrophages . The 50% lethal dose for guinea pigs infected with 25 degrees C-grown F889 was log10 5.0 CFU and that for 41 degrees C-grown F889 was log10 5.7 CFU, or a fivefold difference . Guinea pig alveolar macrophages were infected in quadruplicate with log10 3.8 CFU of F889 cells grown at either 25 or 41 degrees C . Counts of F889 in the alveolar macrophages infected with 25 degrees C-grown bacteria were 40% greater after 1 day of incubation (P = 2 X 10(-4)) than were counts in the alveolar macrophage suspensions inoculated with 41 degrees C-grown bacteria . However, the counts were not significantly different after 3 days of incubation . Examination of cover slip cultures of guinea pig alveolar macrophages infected with 25 degrees C-grown or 41 degrees C-grown bacteria showed that the bacteria grown at the lower temperature were twice as likely to be macrophage-associated after 1 h of incubation than were the bacteria grown at the higher temperature . Growth at the lower temperature was also associated with a change in reactivity with monoclonal antibodies, but not with a change in plasmid content . Thus, environmental temperature may play an important role in modulating the virulence of L . pneumophila, possibly by affecting bacterial adherence to host cells. EMBO J, 1987 Nov, 6(11), 3515 - 20 Biological consequences of segmental alterations in mRNA stability: effects of deletion of the intercistronic hairpin loop region of the Rhodobacter capsulatus puf operon; Klug G et al.; It has been proposed that intercistronic stem and loop structures located in the puf operon of the photosynthetic bacterium Rhodobacter capsulatus account for segmental differences in transcript stability and consequently, the differential expression of the B870 and reaction center (RC) proteins encoded by puf . We report here that deletion of these structures leads to a failure to detect as discrete fragments the B870-encoding 0.49 kb and 0.50 kb mRNA segments located upstream from the site of the hairpins . The absence of these stable transcript fragments is associated with altered stoichiometry of the B870 and RC pigment-protein complexes in the bacterial intracytoplasmic membrane and a decreased rate of cell growth under photosynthetic conditions . These results support the view that the hairpin loop structures of the puf intercistronic region function in vivo to impede exoribonucleolytic degradation of upstream mRNA and establish that segmental variations in mRNA stability have a biologically important role in regulating the expression of puf operon genes. J Exp Med, 1987 Nov 1, 166(5), 1377 - 89 Phagocytosis of Legionella pneumophila is mediated by human monocyte complement receptors; Payne NR et al.; We have examined receptors mediating phagocytosis of the intracellular bacterial pathogen, Legionella pneumophila . Three mAbs against the type 3 complement receptor (CR3), which recognizes C3bi, inhibit adherence of L . pneumophila to monocytes by 64 +/- 8% to 74 +/- 11% . An mAb against the type 1 complement receptor (CR1), which recognizes C3b, inhibits adherence by 68 +/- 1% . mAbs against other monocyte surface antigens do not significantly influence adherence . Monocytes plated on substrates of L . pneumophila membranes modulate their CR1 and CR3 receptors but not Fc receptors; such monocytes bind 70% fewer C3b-coated erythrocytes and 53% fewer C3bi-coated erythrocytes than control monocytes . Adherence of L . pneumophila to monocytes in nonimmune sera is dependent on heat-labile serum opsonins; adherence is markedly reduced in heat-inactivated serum (84% reduction) or buffer alone (97% reduction) compared with fresh serum . mAbs against CR1 and CR3 receptors also inhibit L . pneumophila intracellular multiplication and protect monocyte monolayers from destruction by this bacterium . This study demonstrates that human monocyte complement receptors, CR1 and CR3, mediate phagocytosis of L . pneumophila . These receptors may play a general role in mediating phagocytosis of intracellular pathogens. Microbiol Sci, 1987 Nov, 4(11), 338 - 41 Temporal and spatial regulation of differentiation in Caulobacter crescentus; Newton A; Asymmetric cell division in the aquatic bacterium C . crescentus has proved to be a fruitful model for the study of cell differentiation . An understanding of the temporal and spatial mechanisms responsible for complex developmental programmes leading to polar morphogenesis is possible in these cells using a combination of genetic, biochemical, and molecular approaches. Biochemistry, 1987 Oct 6, 26(20), 6521 - 6 Purification and partial characterization of the membrane-bound cytochrome o(561,564) from Vitreoscilla; Georgiou CD et al.; Cytochrome o(561,564) terminal oxidase was solubilized from the membrane fraction of the bacterium Vitreoscilla sp., strain C1, and purified by differential pH dialysis, gel filtration chromatography, and ion-exchange chromatography . Subunit molecular weights, determined on sodium dodecyl sulfate-polyacrylamide gels by the Ferguson plot method, were 49,500 and 23,500 . There were two protohemes IX, two coppers, and 45 mol of phosphorus per mole of protomer (73,000) . The molecular weight of the cytochrome o complex estimated by chromatography on Sephacryl-400 in deoxycholate was 265,000, which is consistent with the enzyme complex under these conditions being a dimer (146,000) with the remaining molecular weight contribution arising from bound phospholipid, deoxycholate, and possibly other, smaller subunits . Difference spectra of the dithionite-reduced enzyme have split alpha absorption maxima at 561 and 564 nm at room temperature and 558 and 561 nm at 77 K . The CO difference spectrum at room temperature has absorption maxima at 570, 534, and 416 nm . Dissociation constants for CO and cyanide binding to the reduced and oxidized forms of the oxidase are 5.2 microM and 3.5 mM, respectively . The hemes in the cytochrome are one electron accepting centers, both with midpoint potentials around +165 mV at pH 7.0 . The enzyme is highly autoxidizable, and its menadiol oxidizing activity is stimulated by phospholipids. Int J Radiat Biol Relat Stud Phys Chem Med, 1987 Oct, 52(4), 603 - 13 Major E . coli heat-stress protein do not translocate: implications for cell survival; Yatvin MB et al.; When Escherichia coli are exposed to heat stress, the majority of proteins in the process of synthesis at the time of heat stress are rapidly translocated to the outer membrane of the bacterium . The synthesis of most of these proteins appears to take place on membrane-bound polyribosomes . With the temperature shift, overall protein synthesis is inhibited while the synthesis of a small group of proteins is initiated . These proteins are not translocated, but remain in the cytosolic compartment, and they are identifiable as heat-stress proteins . Both the translocation phenomenon and the retention of heat-stress proteins in the cytosolic compartment in proximity to the nucleoid could counteract the effects of heat stress . The translocated proteins may operate by stabilizing the outer membrane prior to the induction of heat-stress proteins and the latter, which are confined to the cytoplasmic compartment, may serve to protect the integrity of the nucleoid structures. J Cell Biol, 1987 Oct, 105(4), 1821 - 8 Localizing the subunit pool for the temporally regulated polar pili of Caulobacter crescentus; Smit J; The pili of the stalked bacterium Caulobacter crescentus are assembled at a specific time in the life cycle at one pole of the cell and are composed of the monomer protein, pilin . A previous study demonstrated that the onset of pilin synthesis occurs well before pili appear on the surface, suggesting that pilin accumulates within the cell . In the present study, an electron microscope immunocytochemistry assay was used to determine the subcellular location of this unassembled pilin and its fate during pilus assembly and cell division . Populations of synchronously growing cells were embedded in epoxy resin at selected times during the cell cycle . Ultrathin sections were treated with pilin-specific antibody, followed by protein A coupled to colloidal gold . It was determined that the cellular location for unassembled pilin was the cell cytoplasm . All cell membranes and regions of nuclear material were poorly labeled . Quantitation demonstrated that label density increased during the period of pilin synthesis and declined during the period of pilus assembly and maintenance . The pilin pool was not unequally segregated at division; e.g., to the daughter cell that is elaborating pili . Mutants which have simultaneously lost the ability to produce flagella, pili, and other polar organelles, possibly due to alterations in the specialized region of polar organelle assembly, were also examined by the immunocytochemistry technique . There was no significant difference in the pilin pool size relative to the wild type, indicating that pilin synthesis continues in the absence of a functioning assembly site . This pattern of synthesis and assembly for the pilus is significantly different from that of the polar flagellum which is produced at the same time and location on the cell surface . These findings are discussed in relation to the hypothesized organization center at the cell pole which may have a major role in directing the assembly of all the polar structures. Biochim Biophys Acta, 1987 Sep 25, 921(2), 275 - 80 Enzymes involved in the formation of 3 beta, 7 beta-dihydroxy-12-oxo-5 beta-cholanic acid from dehydrocholic acid by Ruminococcus sp . obtained from human intestine; Akao T et al.; Ruminococcus sp . PO1-3 from human intestinal flora reduced dehydrocholic acid to 3 beta-hydroxy-7,12-dioxo-5 beta-cholanic acid by means of the enzyme 3 beta-hydroxysteroid dehydrogenase (Akao, T., Akao, T., Hattori, M., Namba, T . and Kobashi, K . (1986) J . Biochem . (Tokyo) 99, 1425-1431) . This bacterium and its crude extract gave rise to another product, showing a lower RF value on TLC, from dehydrocholic acid . The product was identified as 3 beta, 7 beta-dihydroxy-12-oxo-5 beta-cholanic acid . The crude extract reduced 7-ketolithocholic acid and its methyl ester, but not 6-ketolithocholic acid and 12-ketochenodeoxycholic acid, in the presence of NADPH, and oxidized ursodeoxycholic acid and beta-muricholic acid, but not cholic acid, chenodeoxycholic acid, deoxycholic acid and hydrocholic acid, in the presence of NADP+ . Therefore, besides 3 beta-hydroxysteroid dehydrogenase, 7 beta-hydroxysteroid dehydrogenase was shown to be present in this bacterium . The two dehydrogenases were clearly separated from each other by butyl-Toyopearl 650 M column chromatography . From dehydrocholic acid, 7 beta-hydroxy-3,12-dioxo-5 beta-cholanic acid was produced by 7 beta-hydroxysteroid dehydrogenase and 3 beta, 7 beta-dihydroxy-12-oxo-5 beta-cholanic acid was produced by combination of two enzymes, 7 beta- and 3 beta-hydroxysteroid dehydrogenase. FEBS Lett, 1987 Sep 14, 221(2), 299 - 304 Phenylhydrazine as probe for cofactor identification in amine oxidoreductases . Evidence for PQQ as the cofactor in methylamine dehydrogenase; van der Meer RA et al.; Homogeneous methylamine dehydrogenase (primary-amine:(acceptor) oxidoreductase (deaminating), EC 1.4.99.3, MADH) from the bacterium Thiobacillus versutus was treated with the inhibitor phenylhydrazine (PH) . Derivatization of the cofactor in MADH took place in a fast reaction to give compound I . A different product, compound II, was formed in a slow reaction at high O2 concentrations . The compounds I and II could be removed from the protein by proteolysis with pronase and purified to homogeneity . Products showing identical absorption spectra and chromatographic behaviour were isolated from the reaction mixture after incubating pyrroloquinoline quinone (PQQ) with PH . Upon dissolving in dimethyl sulphoxide, both the enzyme-derived as well as the model-system-derived compounds I and II were nearly quantitatively transformed into PQQ . The conclusion is, therefore, that MADH from T . versutus contains covalently bound PQQ, removable from the protein with pronase, and not a structural analogue of this cofactor without the carboxylic acid groups, as was recently proposed for MADH from Bacterium W3A1 {(1986) Biochem . Biophys . Res . Commun . 141, 562-568} . The properties of compounds I and II suggest that they are the 'azo adduct' and the 'hydrazone adduct' of PH and PQQ at the C(5)-position, respectively . The finding that the reaction of a hydrazine with PQQ can lead to two different products, in enzymes as well as in a model system, has important implications for the interpretation of recent comparative studies aimed at detection of PQQ in amine oxidoreductases with Raman spectroscopy. J Biol Chem, 1987 Sep 5, 262(25), 12096 - 103 Non-adenylylated bis(5'-nucleosidyl) tetraphosphates occur in Saccharomyces cerevisiae and in Escherichia coli and accumulate upon temperature shift or exposure to cadmium; Coste H et al.; A new set of bis(5'-nucleosidyl) tetraphosphates, the Bp4B' nucleotides (B and B' = C, G, or U not equal to A), are demonstrated in living cells . In exponentially growing Saccharomyces cerevisiae, cellular concentrations of Cp4U, Up4U, Gp4G, Cp4C, Gp4U, and Gp4C are 210, 200, 60, 50, 40, and 30 nM, respectively . It is likely that these nucleotides originate from the action of diadenosine-5',5"'-P1,P4-tetraphosphate alpha,beta-phosphorylase, an enzyme recently found in yeast . Upon temperature shift or exposure to cadmium, the Bp4B' nucleotides strongly accumulate in the yeast cells . In Escherichia coli, the same nucleotides occur, and similar effects of temperature shift or of cadmium are observed . However, in the bacterium, the origin of these nucleotides is not known . To quantitate these nucleotides in cellular extracts, specific procedures were developed . In the first step, after purification of the mixture of Np4N' (N and N' = A, C, G, or U) nucleotides, the Ap4N nucleotides are specifically removed by incubation with lysyl-tRNA synthetase . In the second step, the Bp4B' species are resolved with the help of anion-exchange high performance liquid chromatography . In the third step, the concentration of each Bp4B' is measured using three coupled enzymatic reactions to produce ATP and bioluminescence . With this strategy, 0.01 pmol of any Bp4B' nucleotide can be reliably detected. Infect Dis Clin North Am, 1987 Sep, 1(3), 595 - 614 Legionella infections; Ching WT et al.; Legionnaires' disease is a distinct clinical entity caused by Legionella pneumophila . Following an epidemic of pneumonia in Philadelphia in 1976, it was found that the bacterium had in fact been first isolated in 1947 . Other species of Legionella have been identified, many of which are indistinguishable from L . pneumophila infection . Legionella species also cause extrapulmonary infections and a mild nonpneumonic form of disease known in its epidemic form as Pontiac Fever. Urologe A, 1987 Sep, 26(5), 256 - 62 {Genital chlamydia infections--clinical aspects, diagnosis and therapy}; Korting HC et al.; Non-gonococcal urethritis and its counterpart in women have become the most frequent genital infection worldwide . As Chlamydia trachomatis is the major causative agent interest has focused on this bacterium . While genital chlamydial infection in men often is manifest the opposite holds true for women . Major complications such as pelvic inflammatory disease can nevertheless turn up . Therefore efficient diagnostic tools are badly needed . If tissue culture procedures are not available direct specimen tests can be performed using fluorescence labelled monoclonal antibodies . For therapy tetracyclines and erythromycin are still the drugs of choice although the cure rates are not totally acceptable . Therefore evaluation of the new quinolones deserves interest. Genetika, 1987 Sep, 23(9), 1708 - 10 {Inhibition of colicin synthesis during integration of transposons into the ColE1 plasmid in Escherichia coli}; Krashennikova LV; Insertions of transposons into ColE1 plasmid have been shown to influence the plasmid-specified colicin synthesis . The quantity of colicin produced by a single bacterium being unchanged, a portion of colicin-producing cells in the population of those containing insertion mutants was 10(1)-10(4)-fold lower than in the case of ColE1 . The effect of transposon was only observed in cis . Insertions were located in different sites of plasmid in both orientations . No secondary DNA rearrangements were formed in regions adjacent to the points of insertions . On the basis of data obtained . It is concluded that the phenomenon can be connected with neither of known types of mutations induced by transposons . A possibility of existence of a new type of such mutations is discussed. J Biol Chem, 1987 Aug 15, 262(23), 11012 - 9 Steady-state kinetic analysis for the reaction of ammonium and alkylammonium ions with methylamine dehydrogenase from bacterium W3A1; McIntire WS; The steady-state kinetic mechanism for the reaction of n-alkylamines and phenazine ethosulfate (PES) or phenazine methosulfate (PMS) with methylamine dehydrogenase from bacterium W3A1 is found to be of the ping-pong type . This conclusion is based on the observations that 1/v versus 1/{methylamine} or 1/{butylamine} plots, at various constant concentrations of an oxidizing substrate, and 1/v versus 1/{PES} or 1/{PMS} plots, at various constant concentrations of a reducing substrate, are parallel . Additionally, the values of kcat/Km for four n-alkylamines are identical when PES is the oxidizing substrate, as were the kcat/Km values for four reoxidizing substrates when methylamine was the reducing substrate . Last, analysis of steady-state kinetic data obtained when methylamine and propylamine are presented to the enzyme simultaneously and PES and PMS are used simultaneously also supports the involvement of a ping-pong mechanism . The enzymic reaction with either methylamine or PES is dependent on the ionic strength, and the data indicate that each interacts with an anionic site on methylamine dehydrogenase . The presence of ammonium ion at low concentration activates the enzyme, but at high concentration this ion is a competitive inhibitor in the reaction involving methylamine and the enzyme . A complete steady-state mechanism describing these ammonia effects is presented and is discussed in light of the nature of the pyrroloquinoline quinone cofactor covalently bound to the enzyme. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1987 Aug, 266(1-2), 116 - 26 Interaction of Escherichia coli and macrophages: alteration by treatment of bacteria with beta-lactam antibiotics; Opferkuch W et al.; Antibiotics are known to exert an influence on the host-parasite relationship either by impairment of immunocompetent cells or by alteration of the bacterium, such as changes of surface properties or the production of toxins . The main problem in investigating the effect of antibiotics on the surface properties of bacteria consists in morphological changes of bacteria (round cell or filament formation) after treatment e.g . with beta-lactam antibiotics . These changes of morphology lead to problems in the comparison of such bacterial forms with untreated organisms . Therefore, in this study outer membrane vesicles from bacteria were used as a model to investigate the effect of antibiotics on the surface properties of Escherichia coli with regard to the interaction with mouse peritoneal macrophages tested by chemiluminescence reaction . It could be shown that these membrane vesicles induce a luminol dependent chemiluminescence response . Treatment of E . coli with different beta-lactams lead to an increase of the stimulating properties . The relative effectiveness of certain antibiotics depended on the particular E . coli strain . Analysis of the different adhesions involved in the stimulation of macrophages revealed that only mannose-sensitive adhesins were increased after treatment with beta-lactam antibiotics . No stimulation of the membrane-bound NAD(P)H-oxidase could be found following the reaction with outer membrane vesicles . Even the treatment of bacteria with antibiotics did not evoke such a reaction. J Periodontol, 1987 Aug, 58(8), 540 - 5 Tissue localization of Actinobacillus actinomycetemcomitans in human periodontitis . II . Correlation between immunofluorescence and culture techniques; Christersson LA et al.; Recent immunohistological studies have suggested that Actinobacillus actinomycetemcomitans is present in the gingival tissues in juvenile periodontitis lesions . The present study examined tissue bound A . actinomycetemcomitans by bacterial culture and immunohistological demonstration of antigen in tissue . A total of 14 periodontitis lesions were examined . Eleven biopsies were obtained from gingiva adjacent to A . actinomycetemcomitans infected pockets, while the remaining three control biopsies were obtained from gingiva adjacent to pockets where subgingival A . actinomycetemcomitans infection could not be detected . Each biopsy was hemisected, one half was used for immunofluorescence microscopic examination while the other half was processed for culture of A . actinomycetemcomitans . The latter section was surface-disinfected, repeatedly washed and then minced to release bacteria from within the tissues . Aliquots from the serial washings and the minced tissue suspension were cultured on medium selective for A . actinomycetemcomitans . Surface disinfection and serial washings gradually decreased cultivable A . actinomycetemcomitans in the washings aliquots . Following tissue disruption, an increase in colony-forming units of A . actinomycetemcomitans was seen from eight of the 11 test biopsies . This bacterium could not be detected in washings or minced tissue suspensions from the control biopsies obtained from lesions in which subgingival A . actinomycetemcomitans was previously not detected . A positive correlation was seen between the presence of A . actinomycetemcomitans antigens in the gingival biopsies and; (1) A . actinomycetemcomitans colony-forming units released from the minced tissues (r = 0.90, p = 0.000), as well as; (2) the colony-forming units from the periodontal pocket (r = 0.62, P = 0.017).(ABSTRACT TRUNCATED AT 250 WORDS) Zentralbl Bakteriol Mikrobiol Hyg {B}, 1987 Aug, 184(6), 495 - 500 {Multiplication and killing temperatures of naturally occurring legionellas}; Schulze-Robbecke R et al.; Suspensions of legionellae and associated bacteria found in a hot water distribution system were maintained in the exponential phase of growth without addition of nutrients by constant agitation and oxygenation and by periodically transferring them to autoclaved tap water . The suspensions were exposed to different temperatures to determine growth and inactivation kinetics of legionellae kept under conditions similar to their natural environment . Multiplication of the legionellae was found to occur in a temperature range between 20 and 43 degrees C and inactivation was observed above 50 degrees C . Decimal reduction times decreased with increasing temperatures . These findings do not support the hypothesis that naturally occurring legionellae are more heat resistant than strains of the bacterium cultured on artificial media . Frequent failure to eradicate legionellae from hot water systems by elevating the water temperature indicates that it is impossible to achieve effective temperature levels concomitantly in all parts of the system. J Bacteriol, 1987 Aug, 169(8), 3801 - 8 Cloning of the gene for myxobacterial hemagglutinin and isolation and analysis of structural gene mutations; Romeo JM et al.; Myxobacterial hemagglutinin (MBHA) is a major developmentally induced protein that accumulates during the period of cellular aggregation in the bacterium Myxococcus xanthus . It has been shown that this lectin is targeted to the cell surface and periplasmic space of developmental cells, suggesting that it may play a role in cell-cell recognition or agglutination . We have cloned the structural gene for MBHA by using synthetic deoxyoligonucleotides containing sequences deduced from the amino acid sequence of MBHA and have used the cloned gene to construct strains of M . xanthus that cannot synthesize MBHA . We found that although the MBHA-deficient strains are delayed in their developmental time course, they are otherwise able to aggregate and sporulate normally . Our results suggest that MBHA may function to increase the efficiency of fruiting-body formation but is not a critical component of cellular aggregation. J Bacteriol, 1987 Aug, 169(8), 3669 - 78 Cloning of the gene for phosphoribulokinase activity from Rhodobacter sphaeroides and its expression in Escherichia coli; Hallenbeck PL et al.; A 3.4-kilobase EcoRI restriction endonuclease fragment has been cloned from the facultatively photoheterotrophic bacterium Rhodobacter sphaeroides and shown to contain the structural gene (prkA) for phosphoribulokinase (PRK) activity . The PRK activity was characterized in Escherichia coli, and the product of the reaction was identified . The prkA gene was localized to a 1,565-base-pair EcoRI-PstI restriction endonuclease fragment and gave rise to a 33-kilodalton polypeptide both in vivo and in vitro . The gene product produced in E . coli was shown to be identical to the gene product produced in R . sphaeroides . The amino acid sequence for the amino-terminal region deduced from the DNA sequence confirmed that derived for partially purified PRK derived from both E . coli and R . sphaeroides . In addition, the 3.4-kilobase EcoRI restriction endonuclease fragment coded for a 37-kilodalton polypeptide of unknown function, and preliminary evidence indicates that this DNA fragment is linked to genes coding for other activities significant in photosynthetic carbon assimilation . The genetic organization and proposed operon structure of this DNA fragment are discussed. J Immunol, 1987 Jul 15, 139(2), 551 - 6 Cytolytic activity of human peripheral blood leukocytes against Legionella pneumophila-infected monocytes: characterization of the effector cell and augmentation by interleukin 2; Blanchard DK et al.; The present study was an in vitro attempt to define the effector mechanisms against the intracellular bacterium Legionella pneumophila . Monocytes from human peripheral blood leukocytes (PBL) were infected in vitro with L . pneumophila and cultured for 2 days to allow intracellular replication of the bacterium . Cells were then labeled with 51Cr and used as targets in a 4-h 51Cr-release assay . We report here that autologous nonadherent PBL effectively lysed infected monocytes, and this activity was enhanced when the effector cells were precultured with IL 2 for 2 days . The IL 2-activated killer cells were also cytolytic against uninfected cultured monocytes, but cytotoxicity was higher against Legionella-infected target cells in a dose-dependent manner . The effector cells were located in Percoll density fractions that were enriched for large granular lymphocytes . The phenotype of the effector cell activated by IL 2 was determined to be OKM1+, OKT11+, partially Leu-11+, and negative for Leu-M1, OKT4, OKT8, and Leu-7, indicating that it is neither a T cell nor a monocyte, and is possibly and NK subset that is Leu-11+ and Leu-7- . Cold target inhibition studies indicated that a similar recognition structure is shared by both infected and uninfected monocytes, but differs from that on K562 tumor target cells . Thus, in addition to tumor surveillance and controlling viral infections, killer cells can be activated to provide protection against intracellular bacterial infections. FEBS Lett, 1987 Jul 13, 219(1), 244 - 8 Electron paramagnetic resonance and magnetic circular dichroism studies of a hexa-heme nitrite reductase from Wolinella succinogenes; Blackmore RS et al.; The nature of the heme centers in the hexa-heme dissimilatory nitrite reductase from the bacterium Wolinella succinogenes has been investigated with EPR and magnetic circular dichroism spectroscopy . The EPR spectrum of the ferric enzyme is complex showing, in addition to magnetically isolated low-spin ferric hemes with g values of 2.93, 2.3 and 1.48, two sets of signals at g = 10.3, 3.7 and 4.8, 3.21, which we assign to two pairs of exchange coupled hemes . The MCD spectra show that the isolated hemes are bis-histidine coordinated and that there is one high-spin ferric heme . The exchange coupling is lost on treatment with SDS. Avian Dis, 1987 Jul-Sep, 31(3), 597 - 600 Lack of protection against Bordetella avium in turkey poults exposed to B . avium-like bacteria; Jackwood MW et al.; Six laboratory experiments were designed to determine whether poults infected with the nonpathogenic Bordetella avium-like (BAL) bacteria would develop immunity to B . avium (BA), the causative agent of turkey coryza . The BAL bacteria were isolated from poults given that organism, but few colonies were observed by 3 weeks postexposure . No serum-agglutinating antibody to the BAL bacterium was detected in poults exposed to that organism . Poults exposed to BAL bacteria either once or twice at different ages were not protected from infection or disease following experimental challenge between 1 and 7 weeks of age with pathogenic BA. Can J Microbiol, 1987 Jul, 33(7), 642 - 6 Effects of temperature upon the cell-free translation system from Coxiella burnetii; Donahue JP et al.; The rate and extent of coliphage Q beta RNA translation by cell-free extracts prepared from Coxiella burnetii were studied . When translations were conducted at temperatures elevated above 37 degrees C, both polypeptide elongation and frequency of initiation were by comparison increased . The ratios of products synthesized from the polycistronic phage mRNA also changed upon increases in translation temperature, especially at 45 degrees C . Although the organism is a moderate acidophile, initiation of protein synthesis in extracts did not occur below pH 6.2, and was superior when the pH was 6.8-7.2 . The results are discussed in context with the known physiological characteristics of this obligate intracellular bacterium. Biochem J, 1987 Jul 1, 245(1), 139 - 43 Structure of two new aminophospholipids from Methanobacterium thermoautotrophicum; Kramer JK et al.; The methanogenic bacterium Methanobacterium thermoautotrophicum (A.T.C.C . 29183) was shown to contain two new aminophospholipids . These are 2-aminoethyl phosphate ester of diphytanylglycerol diether and a sugar containing bisdiphytanyldiglycerol tetraether . The two aminophospholipids were stable to acid methanolysis except for the sugar on the bisdiphytanyldiglycerol tetraether . Strong acid (6 M-HCl) hydrolysed the alkyl ether and aminophosphate ester bonds . The structure of the phosphate linkage was demonstrated by 31P n.m.r., and the 2-ethanolamine structure was elucidated by 1H- and 13C-n.m.r . spectroscopy and by fast-atom-bombardment m.s. J Bacteriol, 1987 Jul, 169(7), 3076 - 81 Substitution of Co alpha-(5-hydroxybenzimidazolyl)cobamide (factor III) by vitamin B12 in Methanobacterium thermoautotrophicum; Stupperich E et al.; Methanobacterium thermoautotrophicum grown on mineral medium contains 120 nmol of Co alpha-(5-hydroxybenzimidazolyl)cobamides (derivatives of factor III) per g of dry cell mass as the sole cobamide . The bacterium assimilated several corrinoids and benzimidazole bases during autotrophic growth . The corrinoids were converted into factor III; however, after three transfers in 5,6-dimethylbenzimidazole (200 microM)-supplemented mineral medium, derivatives of factor III were completely replaced by derivatives of vitamin B12, which is atypical for methanogens . The total cobamide content of these cells and their growth rate were not affected compared with factor III-containing cells . Therefore, the high cobamide content rather than a particular type of cobamide is required for metabolism of methanogens . Derivatives of factor III are not essential cofactors of cobamide-containing enzymes from methanogenic bacteria, but they are the result of a unique biosynthetic ability of these archaebacteria . The cobamide biosynthesis include unspecific enzymes, which made it possible either to convert non-species-derived corrinoids into derivatives of factor III or to synthesize other types of cobamides than factor III . The cobamide biosynthesis is regulated by its end product . In addition, the uptake of extracellular cobamides is controlled, and the assimilated corrinoids regulate cellular cobamide biosynthesis. Mol Gen Mikrobiol Virusol, 1987 Jul, (7), 11 - 3 {Use of the plasmid R89S replicon for constructing integration vectors}; Zinchenko VV et al.; It has been shown that the plasmid R89S derivatives can be used as integrative vectors for bacteria in which the plasmid is unable to replicate autonomously . The chromosomal and plasmid fragments of phototrophic bacterium Rhodobacter sphaeroides have been cloned in plasmid pVZ365, a SmRKmR-derivative of R89S . The obtained recombinant plasmids were mobilized into R . sphaeroides cells by the I pcP-group conjugative plasmid R751 . The frequencies of the SmR-transconjugants formation are 3.7.10(-5) to 5.6.10(-3) per recipient cell . The formation of the SmR-transconjugants has not been revealed in case of the plasmid pVZ365 mobilization . The recombinant molecules containing R . sphaeroides plasmid fragments have been shown to integrate into endogenous plasmids and form cointegrates with them. J Clin Periodontol, 1987 Jul, 14(6), 370 - 2 Cell surface hydrophobicity of Actinobacillus actinomycetemcomitans Y4; Kozlovsky A et al.; Oral bacteria colonize the dento-gingival tissues in a selective manner . Hydrophobic reactions have been suggested as one of the major mechanisms of adhesion . Hydrophobicity of Actinobacillus actinomycetemcomitans Y4 (Aa) cells was studied in vitro using adherence to the liquid hydrocarbon, octane . Adherence of Aa cells to octane varied from 60-90%, depending on the medium in which they were grown, age of the culture and the buffer in which the assay was carried out . These data suggest that Aa is a hydrophobic bacterium, the hydrophobicity of which is expressed to a varying degree, and may have a role in its adherence to oral tissues. Infect Immun, 1987 Jul, 55(7), 1607 - 9 Evidence for sialyl glycoconjugates as receptors for Bordetella bronchiseptica on swine nasal mucosa; Ishikawa H et al.; The nature of the receptors for Bordetella bronchiseptica was investigated by using the in vitro adherence assay system . The results indicated that sialyl glycoconjugates acted as receptors on swine nasal mucosa . These results were obtained by two independent approaches: inhibition of epithelial cell adherence with sialic acid-containing compounds but not with compounds lacking sialic acid residues and loss of adherence after treatment of epithelial cells with periodate or neuraminidase . B . bronchiseptica seems to have strong affinity for mucin . This may help the bacterium to colonize the mucosal surfaces of the swine nasal cavity. J Bacteriol, 1987 Jul, 169(7), 3035 - 43 Amino acid concentrations in Rhodospirillum rubrum during expression and switch-off of nitrogenase activity; Kanemoto RH et al.; The amino acid concentrations in the phototrophic bacterium Rhodospirillum rubrum were measured during growth under nif-repressing and nif-derepressing conditions . The effects of ammonium, glutamine, darkness, phenazine methosulfate, and the inhibitors methionine sulfoximine and azaserine on amino acid levels of cells were tested . The changes were compared to changes in whole-cell nitrogenase activity and ADP-ribosylation of dinitrogenase reductase . Glutamate was the dominant amino acid under every growth condition . Glutamine levels were equivalent when cells were grown on high-ammonia (nif-repressing) medium or glutamate (nif-derepressing) medium . Thus, glutamine is not the solitary agent that controls nif expression . No other amino acid correlated with nif expression . Glutamine concentrations rose sharply when either glutamate-grown or N-starved cells were treated with ammonia, glutamine, or azaserine . Glutamine levels showed little change upon treatment of the cells with darkness or ammonium plus methionine sulfoximine . Treatment with phenazine methosulfate resulted in a decrease in glutamine concentration . The glutamine concentration varied independently of dinitrogenase reductase ADP-ribosylation, and it is concluded that an increase in glutamine concentration is neither necessary nor sufficient to initiate the modification of dinitrogenase reductase . No other amino acid exhibited changes in concentration that correlated consistently with modification . Glutamine synthetase activity and nitrogenase activity were not coregulated under all conditions, and thus the two regulatory cascades perceive different signal(s) under at least some conditions. Biochem Biophys Res Commun, 1987 Jun 15, 145(2), 868 - 75 Spectral properties of nitric oxide complex of cytochrome c' from Rhodopseudomonas capsulata B100; Yoshimura T et al.; The spectral properties for NO complexes of ferric and ferrous cytochrome c' from photosynthetic bacterium Rhodopseudomonas capsulata B100 are reported . The electronic absorption, MCD, and EPR spectra have been compared with those of the NO complexes of the other cytochromes c' and horse heart cytochrome c . The NO-ferrous cytochrome c' would be a mixture of NO complexes with six- and five-coordinate nitrosylheme, suggesting that the heme-iron to histidine bond in the ferrous cytochrome c' is more stable than that from chemoheterotrophic bacteria . The reaction product of ferric cytochrome c' with NO exhibited the spectra similar to NO-ferric derivatives of the other hemoproteins, which indicates the formation of NO-ferric cytochrome c'. Mol Gen Genet, 1987 Jun, 208(1-2), 152 - 8 Effect of UTP and GTP pools on attenuation at the pyrE gene of Escherichia coli; Poulsen P et al.; We have used the galK gene, minus its promoter, to quantitate transcription of the orfE--pyrE operon of Escherichia coli in front of and after the intercistronic attenuator . Expression of the hybrid genes was studied in a bacterium with mutations that permit changes in the UTP and GTP pools during exponential growth . It was found that the greater part of pyrE gene regulation by the nucleotides takes place at the intercistronic attenuator and that promoter control contributes only little, ca . twofold . When pools of both UTP and GTP were high only 5%-6% of the mRNA chains were continued into the pyrE gene . However, when the UTP pool was reduced (from 1.3 to 0.2 mumol/g dry weight) nearly 100% of transcription passed the attenuator . Likewise, a reduction in the GTP pool (from 3.2 to 0.8 mumol/g dry weight) resulted in 25%-30% escape of attenuation . Regulation by attenuation disappeared when a premature stop-codon was introduced near the end of orfE such that translational coupling to transcription was prevented in the attenuator area . Therefore, we attribute the modulation of attenuation to nucleotide-induced variations in the kinetics of mRNA chain elongation . In support for this it was found that an RNA polymerase mutant with reduced RNA chain growth rate transcribed past the pyrE attenuator at a high frequency in the presence of a high UTP pool, but only when coupling of translation to transcription was allowed at the end of orfE. J Bacteriol, 1987 Jun, 169(6), 2667 - 74 Nucleotide sequence of a gene cluster involved in entry of E colicins and single-stranded DNA of infecting filamentous bacteriophages into Escherichia coli; Sun TP et al.; Mutations in fii or tolA of the fii-tolA-tolB gene cluster at 17 min on the Esche