Biochem J, 1985 Aug 1, 229(3), 663 - 8 Transcription of Rhodospirillum rubrum atp operon; Falk G et al.; The photosynthetic non-sulphur bacterium Rhodospirillum rubrum contains a cluster of five genes encoding the subunits of F1-ATPase {Falk, Hampe & Walker (1985) Biochem . J . 228, 391-407} . Transcription of these genes has been studied by two methods, transcriptional mapping with S1 nuclease and primer extension analysis . Thereby a 5'-end in RNA derived from this region has been demonstrated at a guanine residue 236 bases before the initiation codon of the gene for the delta-subunit, the first in this cluster . DNA sequences on the 5' side of this nucleotide show some similarity to promoters in Escherichia coli, but are not apparently related to sequences upstream of the Rhodopseudomonas blastica atp operon . A 3'-end in RNA derived from this gene cluster has been demonstrated by S1-nuclease mapping . This is found before a run of thymidylate residues in the DNA, on the 3' side of a region of dyad symmetry . In E . coli these features are characteristic of rho-independent transcriptional termination signals . It appears from these studies and from the organization of the genes that the five genes in the atp cluster may be co-transcribed from this promoter and that transcripts terminate at the region of dyad symmetry. J Biol Chem, 1985 Jul 15, 260(14), 8430 - 7 Branch point control by the phosphorylation state of isocitrate dehydrogenase . A quantitative examination of fluxes during a regulatory transition; Walsh K et al.; To understand how enzymatic pathways respond to changing external conditions, the fluxes through the tricarboxylic acid cycle and ancillary reactions were determined under three different growth conditions in Escherichia coli . The velocities through the major steps in each pathway were measured (a) for growth on acetate alone, (b) for growth on acetate plus glucose, and (c) during the transition caused by addition of glucose to cells growing on acetate . During the transition, the carbon flow through the Krebs cycle decreased by a factor of 5 despite an increase in the growth rate of the culture . Under these conditions, the dephosphorylation of isocitrate dehydrogenase caused a 4-fold increase in its activity . This, together with the decreased rate of substrate production and the kinetic parameters of the branch point enzymes, led to a cessation of the flux through the glyoxylate shunt . The decreased rate of acetyl-CoA turnover, not an inhibition of acetate transport, caused a slower rate of acetate uptake in the presence of glucose . The modulation of protein phosphorylation and metabolite levels is one of the regulatory mechanisms which enables the bacterium to make dramatic shifts between metabolic pathways within a fraction of a doubling time. Jpn J Cancer Res, 1985 Jul, 76(7), 657 - 62 Antitumor activity of Sphaerotilus natans and its slime fraction in mice; Mifuchi I et al.; Antitumor activity of the whole cell and slime of an aquatic sheathed bacterium, Sphaerotilus natans IAM 12068, against ascites form of Ehrlich carcinoma in ddY mice was investigated . Intraperitoneal injection of whole cells and the slime fraction showed remarkable antitumor activity against mice inoculated with 10(4) to 10(5) tumor cells, the slime fraction being more effective . To examine the chemical nature of the active principle in the slime fraction, separation by Sepharose 4B gel filtration was carried out and two fractions designated as GF-P-1 and GF-P-2, which are mainly composed of protein, carbohydrate, and lipid, were obtained . GF-P-1 fraction, which contains large amounts of fucose and unidentified sugar as neutral sugar, showed marked antitumor activity at half the dose of the slime fraction, whereas the antitumor activity of GF-P-2, which is composed mainly of protein, was weak . This finding indicates that GF-P-1 fraction of S . natans slime may be a main active principle . The consistently demonstrable antitumor activity of GF-P-1 was abrogated by treatment of mice with silica, an anti-macrophage agent, suggesting that the antitumor activity of GF-P-1 depends on the activation of macrophages. Nature, 1985 Jun 20-26, 315(6021), 686 - 8 Lattice mobility and anomalous temperature factor behaviour in cytochrome c'; Finzel BC et al.; Atomic temperature factors (B-values) obtained from X-ray refinement experiments provide empirical estimates of protein mobility that have been correlated with both theoretical simulations of protein dynamics and experimental studies of antibody reactivity . The comparison of B-values with protein solution properties requires adjustment of the apparent atomic mobilities to compensate for the effects of the crystal environment . Here we compare crystallographically independent subunits of the dimeric cytochrome c' from the bacterium Rhodospirillum molischianum to examine how lattice effects influence refined B-values . In addition to local effects on protein mobility at crystal contacts, we show that B-value differences up to 12 A between subunits result from lattice disordering effects that approximate to concerted rotations of the molecules about a crystal symmetry axis. Biochem J, 1985 Jun 15, 228(3), 769 - 71 A novel hopanoid, 30-(5'-adenosyl)hopane, from the purple non-sulphur bacterium Rhodopseudomonas acidophila, with possible DNA interactions; Neunlist S et al.; A novel hopanoid, 30-(5'-adenosyl)hopane, was isolated from the purple non-sulphur bacterium Rhodopseudomonas acidophila and identified . The significance of this triterpenoid in terms of bacteriohopanepolyol biosynthesis, membrane reinforcement and possible interactions with nucleic acids is discussed. J Biochem (Tokyo), 1985 Jun, 97(6), 1679 - 88 Intermolecular fructosyl and levanbiosyl transfers by levan fructotransferase of Arthrobacter ureafaciens; Tanaka K et al.; A purified levan fructotransferase preparation from the culture of the bacterium Arthrobacter ureafaciens, which produces di-D-fructose 2,6':6,2' dianhydride (difructose anhydride IV) from levan by an intramolecular levan fructosyl transfer (ILFT) reaction, was found to produce a trioligofructan and a tetraoligofructan from levan in the presence of levanbiose, indicating the intermolecular fructosyl and levanbiosyl transfer (LFT and LBT) reactions . The tri- and tetraoligofructans were identified to be levantriose and -tetraose respectively . Increase in the levanbiose concentration brought about increased production of both oligofructans with decreased formation of difructose anhydride IV, supporting the previous theory proposed by Tanaka et al . (1983) that the ILFT, LFT, and LBT reactions are catalyzed by the same enzyme . In addition, there existed a roughly stoichiometric relationship between the increase and decrease in the productions of these oligofructans, and the LBT reaction was found to occur more intensively than the LFT reaction . Acceptor specificity of the LFT and LBT reactions was studied using fifteen sugars including mono-, di-, and trisaccharides . The enzyme showed both of the reactions only with levanbiose, -triose, and kestose, indicating that the exposed non-reducing levanbiosyl residue was essential for the acceptor and suggesting the existence of a levanbiosyl acceptor site on the enzyme molecule. Proc Natl Acad Sci U S A, 1985 Jun, 82(11), 3616 - 20 Bacterial peptide chain release factors: conserved primary structure and possible frameshift regulation of release factor 2; Craigen WJ et al.; Escherichia coli peptide chain release factors are proteins that direct the termination of translation in response to specific peptide chain termination codons . The mechanisms of codon recognition and peptidyl-tRNA hydrolysis are unknown . We have characterized the genes encoding release factor 1 (RF-1) and release factor 2 (RF-2) to study the structure-function relationships of the proteins and their regulation in the bacterium . In this report, we present the gene structure of RF-1 and RF-2, and a partial peptide sequence of RF-2 . RF-1 and RF-2 are highly homologous in their primary structure . In addition, an in-frame premature opal (UGA) termination codon is located within the RF-2 coding region at amino acid position 26 . This region of the protein was sequenced by automated Edman degradation to confirm the predicted reading frame, and a second independent isolate of the RF-2 gene was identified and sequenced to confirm the DNA sequence . These results imply that a frameshift occurs prior to the premature termination codon, thus allowing for translation of RF-2 to be completed . This may represent a mechanism of translational control of RF-2 expression . An alternative possible means of translational regulation is discussed. Biochem J, 1985 Jun 1, 228(2), 391 - 407 Nucleotide sequence of the Rhodospirillum rubrum atp operon; Falk G et al.; The nucleotide sequence was determined of a 8775-base-pair region of DNA cloned from the photosynthetic non-sulphur bacterium Rhodospirillum rubrum . It contains a cluster of five genes encoding F1-ATPase subunits . The genes are arranged in the same order as F1 genes in the Escherichia coli unc operon . However, as in the related organism Rhodopseudomonas blastica, neither genes for components of F0, the membrane sector of ATP synthase, nor a homologue of the E . coli uncI gene are associated with this locus, as they are in E . coli. Science, 1985 May 31, 228(4703), 1101 - 3 Adaptation of the membrane lipids of a deep-sea bacterium to changes in hydrostatic pressure; DeLong EF et al.; The fatty acid composition of the cell membrane of the barophilic marine bacterium CNPT3 was found to vary as a function of pressure . Greater amounts of unsaturated fatty acids were present in bacteria growing at higher pressures . The results suggest adaptations in the membrane lipids to environmentally relevant pressures . This response to pressure appears to be analogous to temperature-induced membrane adaptations observed in other organisms. Biochim Biophys Acta, 1985 May 31, 807(3), 308 - 19 Soluble cytochrome composition of the purple phototrophic bacterium, Rhodopseudomonas sphaeroides ATCC 17023; Meyer TE et al.; A detailed study of the soluble cytochrome composition of Rhodopseudomonas sphaeroides (ATCC 17023) indicates that there are five c-type cytochromes and one b-type cytochrome present . The molecular weights, heme contents, amino acid compositions, isoelectric points, and oxidation-reduction potentials were determined and the proteins were compared with those from other bacterial sources . Cytochromes c2 and c' have previously been well characterized . Cytochrome c-551.5 is a diheme protein which has a very low redox potential, similar to certain purple bacterial and algal cytochromes . Cytochrome c-554 is an oligomer, which is spectrally similar to the low-spin isozyme of cytochrome c' found in other purple bacteria (e.g., Rhodopseudomonas palustris cytochrome c-556) . An unusual high-spin c-type heme protein has also been isolated . It is spectrally distinguishable from cytochrome c' and binds a variety of heme ligands including oxygen . A large molecular-weight cytochrome b-558 is also present which appears related to a similar protein from Rhodospirillum rubrum, and the bacterioferritin from Escherichia coli . None of the soluble proteins appear to be related to the abundant membrane-bound c-type cytochrome in Rps . sphaeroides which has a larger subunit molecular weight similar to mitochondrial cytochrome c1 and chloroplast cytochrome f. Science, 1985 May 24, 228(4702), 958 - 62 Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine; Young JF et al.; The circumsporozoite (CS) protein of the human malaria parasite Plasmodium falciparum may be the most promising target for the development of a malaria vaccine . In this study, proteins composed of 16, 32, or 48 tandem copies of a tetrapeptide repeating sequence found in the CS protein were efficiently expressed in the bacterium Escherichia coli . When injected into mice, these recombinant products resulted in the production of high titers of antibodies that reacted with the authentic CS protein on live sporozoites and blocked sporozoite invasion of human hepatoma cells in vitro . These CS protein derivatives are therefore candidates for a human malaria vaccine. J Biol Chem, 1985 May 10, 260(9), 5526 - 32 Bordetella pertussis invasive adenylate cyclase . Partial resolution and properties of its cellular penetration; Hanski E et al.; The existence of an invasive adenylate cyclase in dialyzed urea extracts of the bacterium Bordetella pertussis has been suggested recently . Gel filtration of B . pertussis dialyzed urea extract shows that the invasive enzyme constitutes only a small portion of the total adenylate cyclase activity found in the extract . Its size is different than the size of the two peaks of adenylate cyclase activity identified in the extract . Ca2+ is absolutely required for the penetration of the invasive enzyme, it also controls the rate of intracellular cAMP accumulation in human lymphocytes exposed to dialyzed extract . These characteristics may be attributed to the increase in the size of the invasive enzyme as found by gel filtration chromatography of the extract in the absence of Ca2+ . Removal of nonpenetrating adenylate cyclase that adheres to lymphocytes permits a direct assay of the intracellular enzyme . The time course of intracellular accumulation of adenylate cyclase activity is similar to the time course of intracellular accumulation of cAMP, suggesting that the invasive enzyme is rapidly deactivated, but not degraded, since it can be detected upon cell disruption . No appreciable amount of the enzyme is introduced when cells are incubated with extract at 4 degrees C for 120 min, then washed and incubated further at 37 degrees C . Concanavalin A inhibits cAMP accumulation irrespective of the time of its addition, and EGTA prevents penetration of the invasive enzyme even if added 20 min after addition of extract . These findings are different from those observed in other bacterial toxins thought to be internalized by receptor-mediated endocytosis . However, the cellular penetration of B . pertussis adenylate cyclase is cell-selective . It does not occur in human erythrocytes . In addition to human lymphocytes, S49 cyc- murine lymphoma, turkey erythrocytes, and rat oocytes accumulate cAMP in response to B . pertussis extract. Prikl Biokhim Mikrobiol, 1985 May-Jun, 21(3), 422 - 7 {Gas chromatographic determination of amino acid configuration in the optically active stationary phase}; Muratov VB et al.; The synthesis and physico-chemical properties of a novel optically active stationary phase--N-stearoyl-L-valyl-t-butylamide (I) applied for GLC separation of enantiomeric alpha-amino acids are described . The optical purity of the compound (I) is not less than 85% . The efficiency of the phase is shown on analyses of the configuration of amino acids in peptidolipids and glycopeptidolipids produced by the paraffin-oxidizing bacterium Mycobacterium paraffinicum. Infect Immun, 1985 May, 48(2), 417 - 21 Nonimmune binding of equine immunoglobulin by the causative organism of contagious equine metritis, Taylorella equigenitalis; Widders PR et al.; This study identifies nonimmune binding of equine immunoglobulin by the causative organism of contagious equine metritis . Immunoglobulin binding to the bacterium was strongest for immunoglobulin G (IgG) and less for IgM; IgA was not bound . Binding of equine IgG was inhibited by human IgG, but not by IgG of domestic animals . Immunoglobulin binding by the bacterium appeared to be directed towards an epitope in the hinge region of the immunoglobulin molecule. J Immunol, 1985 May, 134(5), 2900 - 7 Site-selective homing of antigen-primed lymphocyte populations can play a crucial role in the efferent limb of cell-mediated immune responses in vivo; Spangrude GJ et al.; Lymphoid cells of the mammalian immune system exhibit special migratory properties within their in vivo environment . This fundamental characteristic is important to the protection of the organism from infection and neoplastic transformation by a process termed immunologic surveillance . The importance of lymphocyte recirculation was addressed by investigating the role of site-selective homing of lymphocytes during the efferent phase of an in vivo adoptively transferred immune response . We approached this problem by using pertussis toxin (PT), a known inhibitor of lymphoid cell migration in vivo . PT is a protein secreted by the bacterium Bordetella pertussis, which induces a selective and long-lasting inhibition of the emigration of lymphocytes from the bloodstream into solid tissue . In this study, we demonstrate that the blockade of lymphocyte extravasation potential mediates inhibition of certain cell-mediated immune responses in vivo . We investigated the effect of PT on the extravasation of antigen-primed lymphocytes into solid tissue . The results show that the loss of lymphocyte homing potential after in vitro treatment of the primed cells with PT is accompanied by an inhibition of antigen-specific contact hypersensitivity after adoptive transfer . This result suggests that the process of lymphocyte extravasation into nonlymphoid tissue sites such as the skin shares fundamental similarities to the selective localization of circulating lymphocytes to lymph nodes . Furthermore, the inhibition of contact hypersensitivity observed after the i.v . adoptive transfer of PT-treated antigen-primed cells could be circumvented by transferring the PT-treated cells directly into a tissue site with the relevant antigen . In contrast, no inhibition in the number of antibody-forming cells present within the spleen was observed after an adoptive transfer of PT-treated sheep red blood cell-primed lymphocytes, a result that is in agreement with radiotracer data . Thus, the inhibitory effect of PT on the adoptive transfer of contact hypersensitivity was established to be a direct result of the PT-mediated alteration of cellular migration, because PT inhibits the entrance of lymphocytes into specific tissue sites without inhibiting other cellular functions . This conclusion is additionally supported by experiments that showed that lymphocytes that have been pretreated with PT exhibit normal in vitro responses, as assayed by mitogenesis in response to concanavalin A and by the differentiation and function of alloantigen stimulated or hapten-specific cytotoxic T lymphocytes.(ABSTRACT TRUNCATED AT 400 WORDS) J Clin Microbiol, 1985 May, 21(5), 838 - 9 Unusual bacterium, group Ve-2, causing peritonitis in a patient on continuous ambulatory peritoneal dialysis; Silver MR et al.; We report a case of peritonitis in a continuous ambulatory peritoneal dialysis patient with an unusual bacterium known as group Ve-2 . This is the first reported case of peritonitis attributable to this organism and only the second well-documented case of infection with this organism in the English literature. J Bacteriol, 1985 May, 162(2), 752 - 5 Presence of 2-methylthioribosyl-trans-zeatin in Azotobacter vinelandii tRNA; Ajitkumar P et al.; Hydroxylated cytokinin, 2-methylthio-N6-(4-hydroxy-3-methylbut-2-enyl) adenosine, was found in the tRNA of Azotobacter vinelandii . This cytokinin had the trans configuration, unlike the cis configuration reported for that from other bacteria . Culture-condition-dependent changes in the content of this thiocytokinin and a few other thionucleosides in the tRNA of this bacterium have been observed. J Dent Res, 1985 May, 64(5), 793 - 8 Rapid identification of periodontal pathogens in subgingival plaque: comparison of indirect immunofluorescence microscopy with bacterial culture for detection of Actinobacillus actinomycetemcomitans; Bonta Y et al.; The sensitivity of indirect immunofluorescence microscopy using specific polyclonal or monoclonal serodiagnostic reagents for Actinobacillus actinomycetemcomitans in subgingival dental plaque ranged from 82-100% as compared with culture on selective or non-selective media . This bacterium was found in 100% of the periodontally diseased sites examined in localized juvenile periodontitis patients and was statistically related to clinical indices of periodontal disease including the Gingival Index, Plaque Index, and Pocket Depth . Indirect immunofluorescence microscopy is a useful technique for the rapid and reliable determination of A . actinomycetemcomitans in human subgingival dental plaque which may be applied to the clinical diagnosis, treatment, and monitoring of periodontitis associated with A . actinomycetemcomitans. Arch Microbiol, 1985 May, 141(4), 273 - 8 The major soluble cytochromes of the obligately aerobic sulfur bacterium, Thiobacillus neapolitanus; Trudinger PA et al.; Four cytochromes were isolated from soluble extracts of the aerobic sulfur bacterium, Thiobacillus neapolitanus . The two most abundant proteins were purified to homogeneity and thoroughly characterized . Cytochrome c-554 (547) is a monomeric, small molecular weight protein which is unusual in having two well-resolved alpha peaks in UV-visible absorption spectra . The redox potential is 208 mV . Native cytochrome c-549 is oligomeric, but has a subunit size of about 26,000 . The yield of this protein could be improved dramatically by washing membranes with 30% ammonium sulfate, but the material solubilized by this method had a larger native molecular weight than that in the initial 0.1 M Tris-Cl extract and behaved differently on chromatography . The properties of cytochrome c-549 including subunit size and UV-visible absorption spectra are similar to mitochondrial cytochrome c1 and chloroplast cytochrome f, which suggests that it may be a modified form of the predominant membrane cytochrome . Based on cytochrome content, it is suggested that T . neapolitanus is not closely related to other thiobacilli. Arch Biochem Biophys, 1985 May 1, 238(2), 373 - 7 Identification of the components of a putative cytochrome bc1 complex in Rhodopseudomonas viridis; Wynn RM et al.; Chromatophore membranes isolated from the bacteriochlorophyll b-containing, photosynthetic purple nonsulfur bacterium, Rhodopseudomonas viridis, have been shown to contain a Rieske iron-sulfur protein, a cytochrome similar to cytochrome c1, and also at least one b-type cytochrome . These observations suggest the presence of a previously undetected cytochrome bc1 complex in this bacterium. J Bacteriol, 1985 May, 162(2), 804 - 9 Molecular and regulatory properties of glutamine synthetase from the phototrophic bacterium Rhodopseudomonas capsulata E1F1; Caballero FJ et al.; The glutamine synthetase of the phototrophic bacterium Rhodopseudomonas capsulata E1F1 was purified to homogeneity by a procedure which used a single affinity chromatography step . Like enzymes from other photosynthetic procaryotes, native glutamine synthetase from R . capsulata E1F1 was found to be a dodecameric protein of approximately 660 kilodaltons with identical subunits of about 55 kilodaltons each . The Stokes radius and S20,w of the native enzyme were 8.35 nm and 19.20, respectively . The enzyme exhibited different aggregation states with detectable oligomers of 1, 2, 3, 4, 6, 8, 10, and 12 subunits . Disaggregation of the glutamine synthetase occurred after the native protein was subjected to electrophoresis in polyacrylamide gels, as well as occurring spontaneously at low ionic strength . Glutamine synthetase from R . capsulata E1F1 was regulated by an adenylylation-deadenylylation mechanism, and the adenylylation state of the protein depended on the nitrogen source, growth phase, and light intensity . Ammonia repressed glutamine synthetase, whereas glycine, serine, alanine, valine, and aspartate were noncompetitive inhibitors of the glutamine synthetase biosynthetic activity. J Bacteriol, 1985 May, 162(2), 584 - 90 DNA transfer occurs during a cell surface contact stage of F sex factor-mediated bacterial conjugation; Panicker MM et al.; Donor bacteria containing JCFL39, a temperature-sensitive traD mutant of the F sex factor, were used at the nonpermissive temperature to accumulate stable mating pairs with recipient cells . At this stage in conjugation, extracellular F pili were removed by treatment with 0.01% sodium dodecyl sulfate . Upon then shifting to the permissive temperature for JCFL39, transfer of the F plasmid was observed . The mating pairs that were accumulated with JCFL39 at the nonpermissive temperature were readily observed by electron microscopy in wall-to-wall contact with the recipient bacteria . These results demonstrate that the traD product, which is known to be required in transferring DNA to a recipient bacterium, acts after the stage at which extracellular F pili are required . In addition, we concluded that DNA transfer takes place while donor and recipient cells are in surface contact and not necessarily through an extended F pilus as envisioned in some models of bacterial conjugation. Am J Vet Res, 1985 Apr, 46(4), 804 - 7 Treatment of acute ocular Moraxella bovis infections in calves with a parenterally administered long-acting oxytetracycline formulation; Smith JA et al.; Acute ocular Moraxella bovis infections were induced in the UV-irradiated eyes of 10 calves . Eight calves developed corneal ulcers in at least 1 eye and were used for the treatment experiment . One randomly selected group of 4 calves with corneal ulcers and M bovis infections in 7 eyes was given a long-acting oxytetracycline formulation in 2 IM dosages of 20 mg/kg of body weight each, 72 hours apart . The other 4 calves with corneal ulcers in 6 eyes and M bovis in all 8 eyes served as nontreated controls . Bilateral ocular cultures were obtained and clinical observations were made daily for 20 days after treatment . After administration of the long-acting drug, new ulcers did not develop in the treated calves, whereas 5 new ulcers developed in the control-group calves during this time . The average durations of increased lacrimation/ulcerated eye were 2 and 12 days after treatment in the treatment and control groups, respectively; the average durations of blepharospasm were 3 and 8 days, respectively . Moraxella bovis was not isolated from any of the eyes of the treatment-group calves for the first 6 days after the antibiotic was administered, but was isolated from 1 eye of 1 treated calf on posttreatment day 7 and daily thereafter, for a total of 14 positive cultures of 160 ocular cultures obtained from the treatment-group calves after treatment . The bacterium was isolated from all eyes and from 144 of 160 cultures from the control-group calves during this time. Appl Environ Microbiol, 1985 Apr, 49(4), 975 - 80 Photoreactivation of UV-irradiated Legionella pneumophila and other Legionella species; Knudson GB; Shortwave UV light was assessed as a feasible modality for the control of Legionnaires disease bacterium in water . The results of this study show that Legionella pneumophila and six other Legionella species are very sensitive to low doses of UV . However, all Legionella species tested effectively countered the germicidal effect of UV when subsequently exposed to photoreactiving light. Arch Biochem Biophys, 1985 Apr, 238(1), 97 - 110 Active transport of nonpolar amino acids in Chromatium vinosum; Cobb AD et al.; The photosynthetic purple sulfur bacterium, Chromatium vinosum, takes up the amino acids, L-phenylalanine and L-leucine, via two apparently different electrogenic, H+/amino acid symports . Na+ serves as an allosteric modulator for leucine transport, lowering the Km for leucine from 66 to 15 microM . C . vinosum cells also contain a system that transports both isoleucine and valine . The isoleucine/valine system has the attributes of a H+/amino acid symport at pH less than 7.5 but appears to function as a H+/Na+ (Li+)/amino acid symport at pH greater than or equal to 7.5 . Na+ gradients produce an allosteric lowering of the Km values for both isoleucine and valine, from 14 to 7 microM and from 34 to 17 microM, respectively . C . vinosum also accumulates D-alanine in an energy-dependent reaction . The transport process appears to involve the electrogenic cotransport of D-alanine and Na+ . The Km value for D-alanine was determined to be 9 microM . Unlike the previously characterized C . vinosum L-alanine/Na+ symport, Na+ gradients did not affect the Km for D-alanine transport . L-Alanine and glycine, but not alpha-aminoisobutyric acid, act as competitive inhibitors for D-alanine transport. J Bacteriol, 1985 Mar, 161(3), 1146 - 55 Differential expression of protein S genes during Myxococcus xanthus development; Downard JS et al.; Protein S, the most abundant protein synthesized during development of the fruiting bacterium Myxococcus xanthus, is coded by two highly homologous genes called protein S gene 1 (ops) and protein S gene 2 (tps) . The expression of these genes was studied with fusions of the protein S genes to the lacZ gene of Escherichia coli . The gene fusions were constructed so that expression of beta-galactosidase activity was dependent on protein S gene regulatory sequences . Both the gene 1-lacZ fusion and the gene 2-lacZ fusion were expressed exclusively during fruiting body formation (development) in M . xanthus . However, distinct patterns of induction of fusion protein activity were observed for the two genes . Gene 2 fusion activity was detected early during development on an agar surface and could also be observed during nutritional downshift in dispersed liquid culture . Gene 1 fusion activity was not detected until much later in development and was not observed after downshift in liquid culture . The time of induction of gene 1 fusion activity was correlated with the onset of sporulation, and most of the activity was spore associated . This gene fusion was expressed during glycerol-induced sporulation when gene 2 fusion activity could not be detected . The protein S genes appear to be members of distinct regulatory classes of developmental genes in M . xanthus. J Reprod Med, 1985 Mar, 30(3 Suppl), 279 - 83 A nonculture test for identification of Chlamydia trachomatis; Amortegui AJ et al.; PIP: The standard isolation test for Chlamydia trachomatis involves growth of C . trachomatis in cycloheximide-treated McCoy cells, but isolation is expensive and complicated and requires special equipment, knowledge, and at least 48 hours . A recent approach to a simple, accurate, and inexpensive test without isolation involves use of enzyme immunoassays . In March 1984, the Chlamydiazyme assay from Abbott Laboratories which detects chlamydial antigens in cervical and urethral swab specimens, was tested in 209 abortion patients in Pittsburgh . 2 swab speicmens were obtained from each patient, 1 to be analyzed for C . trachomatis antigens with Chlamydiazyme and 1 to be used for chlamydial isolation tests . 80% of the patients were white and 70% were unmarried . Their ages ranged from 18-38 years with a mean age of 24.4 years . 72 had had urogenital symptoms within the preceding month . C . trachomatis was isolated in McCoy cells after subculturing from 18 of 209 patients (8.6%) . C . trachomatis antigens were found with Chlamydiazyme in 13 of 18 patients (72.2%) whose specimens yielded the bacterium . The specificity of the enzyme immunoassay procedure was 98.4% (188 of 191) . Overall, 201 of 209 samples (96.2%) were identified correctly with Chlamydiazyme, as compared to isolation after subculture . Of the 18 isolates, only 13 were positive on primary isolation and 5 grew on subsequent subculture . 10 of the 13 primary isolate specimens and 3 of the 5 which grew on subculture were reactive with Chlamydiazyme . The same number of infections was detected with Chlamydiazyme and primary isolation, although only 10 of 13 patients were positive with both methods . 3 specimens were reactive with Chlamydiazyme although organisms were not isolated . 2 of the 3 patients had vaginal discharge . 2 of the 3 were recultured within 6 weeks and both yielded positive Chlamydiazyme results in the absence of C . trachomatis isolation . It was considered unlikely that C . trachomatis would be isolated since all patients had received prophylactic tetracycline treatment after the abortion procedures . Radiobiologiia, 1985 Mar-Apr, 25(2), 258 - 60 {A method of estimating the effectiveness of ionizing radiation and magnetic fields for phage induction in lysogenic bacterial culture}; Anosova MG et al.; Either the difference delta N of the content of free phage particles in the experiment and the control or K ratio of these values can be used to estimate the effectiveness of ionizing radiation or other agents inducing phage formation in a lysogenic bacterium culture . The estimation technique the results of which are nearly independent of the fluctuations in the number of phage particles in the control, the inductor dose being invariable, is the most adequate one . The induction of phage in E . coli K12 (lambda) culture by X-rays and magnetic field is an example illustrating that the K ratio, which can be called "the induction coefficient", is in a good agreement with the requirement mentioned above . A possible nature of the phenomenon observed is discussed. Biochimie, 1985 Mar-Apr, 67(3-4), 343 - 7 The SOS system; d'Ari R; In the bacterium Escherichia coli DNA damaging treatments such as ultraviolet or ionizing radiation induce a set of functions called collectively the SOS response, reviewed here . The regulation of the SOS response involves a repressor, the LexA protein, and an inducer, the RecA protein . After DNA damage an effector molecule is produced--possibly single stranded DNA--which activates the RecA protein to a form capable of catalysing proteolytic cleavage of LexA . The repressors of certain temperate prophages are cleaved under the same conditions, resulting in lysogenic induction . SOS functions are involved in DNA repair and mutagenesis, in cell division inhibition, in recovery of normal physiological conditions after the DNA damage is repaired, and possibly in cell death when DNA damage is too extensive . The SOS response also includes several chromosomal genes of unknown function, a number of plasmid encoded genes (bacteriocins, mutagenesis), and lysogenic induction of certain prophages . DNA damaging treatments seem to induce DNA repair and mutagenic activities and proviral development in many species, including mammalian cells . In general, substances which are genotoxic to higher eukaryotes induce the SOS response in bacteria . This correlation is the basis of the numerous bacterial tests for genotoxicity and carcinogenicity. Eur J Biochem, 1985 Feb 15, 147(1), 47 - 52 Identification of the major human sialoglycoprotein from red cells, glycophorin AM, as the receptor for Escherichia coli IH 11165 and characterization of the receptor site; Jokinen M et al.; The pyelonephritogenic Escherichia coli strain 1 H 11165 specifically agglutinates human erythrocytes carrying the M blood group antigen . The polymorphic forms of this antigen, M and N, are located in the NH2-terminal region of the major human red-cell sialoglycoprotein, glycophorin A . Radioactively labeled glycophorin A from M cells specifically bound to the bacteria . Purified glycophorin AM, but not glycophorin AN, efficiently inhibited for binding . Mild periodate treatment oxidized the NH2-terminal serine in glycophorin AM and this resulted in loss of binding to the bacteria . High concentrations of serine and alkali-labile oligosaccharides derived from glycophorin AM inhibited the binding, whereas the synthetic M-specific NH2-terminal pentapeptide Ser-Ser-Thr-Thr-Gly did not . Neuraminidase treatment of glycophorin AM did not destroy the binding . The most efficient inhibition of binding was observed with the N-terminal glyco-octapeptide obtained from glycophorin AM by CNBr cleavage . This peptide contains both the essential serine residue and the alkali-labile oligosaccharides, which both are recognized by the bacterium. J Bacteriol, 1985 Feb, 161(2), 750 - 7 Three-dimensional architecture of the cell sheath and septa of Methanospirillum hungatei; Shaw PJ et al.; The methanogenic bacterium Methanospirillum hungatei exists as filaments which have a very unusual cell wall architecture, comprising a long cylindrical sheath within which there may be many individual cells arranged in a line . The sheath has a two-dimensional crystalline structure, and the cells are separated within the tube by septa which also have a crystalline structure . We have used computer image processing of tilted-view electron micrographs to analyze the structure in negative stains of both of these components in three dimensions . The repeating unit of the sheath consists of four approximately spherical domains ca . 2.5 nm in diameter arranged in a row . Based on observations of the type of lattice imperfections that occur, we suggest that each of the domains represents a separate polypeptide subunit and that the subunits are incorporated into the wall one by one . The septa are circular plates of remarkably constant size . They are normally found as double layers . They are hexagonally symmetrical and consist of trimerically associated subunits which interact about dimer axes to form an open network containing large pores ca . 15 nm in diameter. J Cell Sci, 1985 Feb, 73, 389 - 98 Isolation and characterization of macronuclei of Paramecium caudatum infected with the macronucleus-specific bacterium Holospora obtusa; Freiburg M; Macronuclei from Paramecium caudatum infected with Holospora obtusa may be isolated on sucrose step gradients . Macronuclei containing primarily infectious forms can be separated from those bearing predominantly reproductive forms . RNA polymerase activity in infected macronuclei is greater by a factor of 5 than that in uninfected macronuclei . Proteinase activity is also significantly higher. Antibiot Med Biotekhnol, 1985 Feb, 30(2), 109 - 13 {Effect of lipopolysaccharide of the pseudotuberculosis bacterium on the functional activity of macrophages}; Kuznetsova TA et al.; Study of the effect of lipopolysaccharide (LPS) of Y . pseudotuberculosis on the culture of peritoneal macrophages of guinea pigs has established that in concentrations of 1-10 micrograms/ml LPS stimulated the absorption and digestive activity of the macrophages . In concentrations of 50-100 micrograms/ml it had a cytotoxic effect on the cell elements and inhibited their phagocytic function . The study of the LPS effect on absorption and liberation of 14C-labeled bacteria of Y . pseudotuberculosis showed that the biopolymer increased 2-3 times the uptake of the labeled antigen from the macrophages and lowered its elimination . In the experiments on mice infected with a labeled culture of S . aureus no increase in the uptake under the effect of the pseudotuberculosis LPS was observed, whereas an increase in the rate of the label elimination was stated . The difference was statistically significant . The results of the experiments are indicative of the specific effect of LPS . The study demonstrated a significant role of LPS of Y . pseudotuberculosis in the pathogenesis and protection of the host from this infection. J Exp Med, 1985 Feb 1, 161(2), 409 - 22 Isolation and characterization of the cytoplasmic and outer membranes of the Legionnaires' disease bacterium (Legionella pneumophila); Gabay JE et al.; Legionella pneumophila, the etiologic agent of Legionnaires' disease, is phagocytized in an unusual way and multiplies in human mononuclear phagocytes in a novel phagosome . As a first step toward understanding these L . pneumophila-phagocyte interactions, we have studied the envelope of L . pneumophila Philadelphia 1 strain . We isolated cell envelopes by treating whole bacterial cells with lysozyme and EDTA to convert them to spheroplasts, then lysing the spheroplasts osmotically or sonically . We resolved the cell envelopes into two membrane fractions by isopycnic centrifugation . We localized NADH oxidase to the fraction of buoyant density 1.145, which we designated cytoplasmic membrane, and lipopolysaccharide (LPS) to the fraction of density 1.222, which we designated outer membrane . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the L . pneumophila outer membrane contains a single major protein species migrating at 28,000 mol wt; this is the major protein of the bacterium . The cytoplasmic membrane also contains a single major protein species migrating at 65,000 mol wt . Surface iodination of the bacteria and agglutination and immunofluorescence studies with rabbit antibody produced against the purified major outer membrane protein (MOMP) revealed that this protein is exposed at the cell surface . We isolated LPS from L . pneumophila membranes by SDS-EDTA treatment . The pattern obtained by subjecting the LPS to SDS-PAGE and staining the gel with silver nitrate suggests that L . pneumophila LPS might be atypical . We studied patient serologic responses to cell envelope components of L . pneumophila Philadelphia 1, a serogroup 1 organism . Sera from patients with evidence of infection with serogroup 1 L . pneumophila contained large amounts of antibody to this strain . Few of these antibodies recognized the MOMP of L . pneumophila . In contrast, greater than 98% of these antibodies were directed against the LPS . This indicates that LPS is the dominant serogroup antigen and the major antigen responsible for the reactivity of patient sera in the indirect fluorescent antibody assay, currently the principal diagnostic assay for Legionella infection. J Bacteriol, 1985 Feb, 161(2), 810 - 2 Induction of chloramphenicol and tetracycline resistance in Flexibacter sp . strain FS-1; Barcak GJ et al.; The gliding bacterium Flexibacter sp . strain FS-1 exhibits inducible resistance to chloramphenicol (Cmr) and tetracycline (Tcr) . Either chloramphenicol or tetracycline alone induced a Cmr Tcr phenotype . The resistance is apparently not plasmid encoded. Biochem Biophys Res Commun, 1985 Jan 31, 126(2), 698 - 704 Heterologous hybridization of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) restores the enzyme activities; Incharoensakdi A et al.; The catalytic core (A8) and small subunit (B) of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) were isolated from two species of cyanobacteria (Aphanothece halophytica and Synechococcus ACMM 323) as well as from the photosynthetic purple sulfur bacterium, Chromatium vinosum . The subunit B is essential for the activity of all three enzymes . The heterologous hybridization of RuBisCO molecules from the three organisms was attempted and the reconstitution of the catalytically active hybrid was achieved between A8 derived from either Aphanothece or Synechococcus and subunit B from Aphanothece, Synechococcus or Chromatium . However, reconstitution of the enzymically active hybrid between A8 from Chromatium and B subunits from the cyanobacteria could not be achieved . Experiments by using high performance liquid column chromatography also showed the formation of a heterologous hybrid possessing RuBP carboxylase activity. Biochem Biophys Res Commun, 1985 Jan 31, 126(2), 685 - 91 Evidence of tyrosine kinase activity in the photosynthetic bacterium Rhodospirillum rubrum; Vallejos RH et al.; The photosynthetic bacterium Rohodospirillum rubrum evidenced tyrosine protein phosphorylation under photoautotrophic conditions in the presence of {32P}Pi . The stability to alkaline treatment of the {32P} bound to the cell-free extract proteins suggested that tyrosine residues were carrying the labelling . One- and two-dimensional high voltage paper electrophoresis analysis revealed that such extracts do contain {32P}-phosphotyrosine residues . Furthermore, the association of alkali stable {32P} bound to specific proteins of the cell-free extract was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis combined with KOH treatment of the gel . A definite argument in favor of protein kinase(s) phosphorylating tyrosine residues in R.rubrum proteins was obtained by partial purification of a tyrosine kinase activity from cell-free extract capable of phosphorylating synthetic peptides that only contain a single tyrosine residue as phosphate acceptor. Biochem J, 1985 Jan 15, 225(2), 441 - 8 The methane mono-oxygenase reaction system studied in vivo by membrane-inlet mass spectrometry; Joergensen L; A membrane-inlet mass spectrometer connected to an open-system cuvette was used for direct measurement of dissolved methane and O2 in bacterial samples of strain OU-4-1, a type II methanotrophic bacterium . A technique was applied for keeping the concentration of dissolved methane or O2 in the sample constant while the concentration of the other dissolved gas was varied . This allowed the reaction mechanism of methane mono-oxygenase to be studied in vivo . The enzyme was found to follow a random bi-reactant mechanism with respect to binding of methane and O2 . Binding of one substrate decreased the affinity for the other . The true binding constants were 1 microM for methane and 0.14 microM for O2 . Studies of HCN inhibition confirmed the random bi-reactant mechanism . HCN was found to be a non-exclusive inhibitor with a binding constant of 0.4 microM. FEBS Lett, 1985 Jan 7, 179(2), 208 - 12 Purification of arogenate dehydrogenase from Phenylobacterium immobile; Mayer E et al.; Phenylobacterium immobile, a bacterium which is able to degrade the herbicide chloridazon, utilizes for L-tyrosine synthesis arogenate as an obligatory intermediate which is converted in the final biosynthetic step by a dehydrogenase to tyrosine . This enzyme, the arogenate dehydrogenase, has been purified for the first time in a 5-step procedure to homogeneity as confirmed by electrophoresis . The Mr of the enzyme that consists of two identical subunits amounts to 69000 as established by gel electrophoresis after cross-linking the enzyme with dimethylsuberimidate . The Km values were 0.09 mM for arogenate and 0.02 mM for NAD+ . The enzyme has a high specificity with respect to its substrate arogenate. Nature, 1985 Jan 3-9, 313(5997), 22 - 7 RNA polymerase heterogeneity in Streptomyces coelicolor; Westpheling J et al.; Two forms of RNA polymerase holoenzyme have been identified in the filamentous differentiating bacterium Streptomyces coelicolor . They contain different species of sigma factor and are distinguishable by their ability to recognize different promoter classes . These and other holoenzyme forms may in part determine the selective expression of different gene sets in this morphologically-complex bacterium. Adv Exp Med Biol, 1985, 189, 107 - 27 Immunological aspects of type 1 and 2 diabetes mellitus; Lernmark A et al.; IDDM occurs predominantly among individuals being class II antigen HLA-DR 3 and/or 4 positive, while NIDDM is not associated with HLA-D . Although the HLA-DR 3 or 4 specificities are prerequisites for IDDM to develop, their high frequencies (about 60%) in the background population preclude tissue typing as a predictive test, underlined by the observation that less than 50% of monozygotic twins are concordant for IDDM . The presence of a number of immune abnormalities argues that the causes of IDDM may be sought in an altered immune reaction against antigens present in the pancreatic B cells and/or in the environment . The majority of IDDM patients of short duration show both cellular and humoral autoimmunity against the pancreatic B cells . Similar phenomena may be observed in patients initially diagnosed as NIDDM and treated with oral hypoglycemic agents . It has been speculated that these patients have a retarded form of IDDM . It is possible that the combination of specific Class II antigen molecule(s) and an invading antigen (virus, bacterium, chemical etc.) presented to the immune system triggers the formation of effector cells such as B lymphocytes and cytotoxic T lymphocytes which also cross-react with the pancreatic B cells . Multiple exposures to this or related antigens throughout several years may eventually lead a sufficient loss of pancreatic B cells to cause insulin dependence. J Lab Clin Med, 1985 Jan, 105(1), 124 - 31 Effect of prior influenza virus infections on susceptibility of AKR/J mice to respiratory challenge with Legionella pneumophila; Berendt RF et al.; Influenza virus administered intranasally to AKR/J mice, followed 3 days later by Legionella pneumophila inoculated intranasally, caused significantly greater mortality than did either of the two agents administered alone . Viable concentrations of both bacteria and viruses dropped in sequentially infected animals, despite the ultimate fatal outcome . Viral concentrations, however, did not decrease as rapidly in sequentially infected as in singly infected mice . Histopathologic lesions were consistent with viral replication aided by elaboration of a bacterial toxin . This observation contrasts with the more commonly observed sequence in which the bacterium proliferates after the virus interferes with host defense . Cell-free preparations were found to have toxic activity. Infect Immun, 1985 Jan, 47(1), 301 - 5 Effect of local and parenteral immunization on implantation of Actinomyces viscosus T6 in rats; Olsson J et al.; Groups of rats immunized in the vicinity of the major salivary glands or immunized intraperitoneally with Actinomyces viscosus T6 and their sham-immunized controls were infected with the homologous bacterium . Significantly higher levels of salivary and serum antibody were induced by intraperitoneal than by salivary gland immunization . There were also significant inverse correlations between the levels of salivary and serum antibody and the levels of implantation of A . viscosus T6 . The level of implantation of A . viscosus T6 was significantly lower in the immunized animals than in the controls . However, antibody had limited capacity to inhibit the establishment of this bacterium. Gene, 1985, 34(2-3), 219 - 26 Broad-host-range plasmid vector for the in vitro construction of transcriptional/translational lac fusions; Nano FE et al.; A broad-host-range plasmid was constructed that allows the in vitro formation of beta-galactosidase fusions . DNA from the photosynthetic bacterium Rhodopseudomonas sphaeroides was cloned into this plasmid and a number of R . sphaeroides isolates were recovered that had varying levels of beta-galactosidase activity . beta-galactosidase antigenic activity from the fusion strains could be localized immunologically in polypeptides with an Mr of 120 000 or greater . Expression of beta-galactosidase activity under control of fusion derivatives was either very low or nonexistent in Escherichia coli relative to R . sphaeroides, indicating that R . sphaeroides promoters or translational start signals function poorly in E . coli. Prep Biochem, 1985, 15(5), 321 - 34 Purification of hydrogenase by fast protein liquid chromatography and by conventional separation techniques: a comparative study; Kovacs KL et al.; Hydrogenase was purified from the photosynthetic bacterium Thiocapsa roseopersicina to homogeneity by various methods . Conventional techniques included separation of the crude protein extract on Phenyl-Sepharose CL 4B, DEAE-cellulose DE52, and chromatofocusing columns or on preparative polyacrylamide gel-electrophoresis . The same protein was isolated by fast protein liquid chromatography (FPLC) in two steps . Comparison of the two different approaches clearly show the superiority of the FPLC method both in enzyme recovery yield and in time requirement. Crit Rev Clin Lab Sci, 1985, 21(4), 323 - 81 Legionella and Legionnaires' disease: a review with emphasis on environmental studies and laboratory diagnosis; Winn WC Jr; Legionella pneumophila and related species are important causes of epidemic bacterial pneumonia and nosocomial infection . This review will discuss this new family of bacteria and the diseases they produce . The classification, general microbiologic characteristics, and ecology of the bacteria will be reviewed and the epidemiology and clinical aspects of the infection will be discussed . More emphasis will be given to issues that are more directly related to laboratory workers and with which the author has had more direct experience: pathology, laboratory diagnosis of human infection, pathogenesis of the infection, and virulence mechanisms of the bacterium . Therapy and prevention of the infection will be discussed more briefly. Arch Oral Biol, 1985, 30(3), 291 - 4 Peptidoglycan from the potentially pathogenic oral bacterium Actinomyces viscosus is a B-cell mitogen; Baker JJ; Cell walls and peptidoglycan from Actinomyces viscosus, strain M-100 were compared for their ability to act as mitogens with spleen cells from germ-free Fischer rats . The cell walls were prepared from trypticase soy broth grown whole cells using a French press, followed by two consecutive washes with 0.1 M tris-HCl buffer, pH 8.0, 1 M NaCl and distilled water . Peptidoglycan was prepared from cell walls by three consecutive formamide extractions at 165 degrees C . On a dry-weight basis, the peptidoglycan was a significantly-better mitogen than cell walls, suggesting that the peptidoglycan is the major mitogenic component of A . viscosus cell walls . Mononuclear spleen cells were separated on a Nylon-wool column into a non-adherent subpopulation enriched for T lymphocytes and a weakly-adherent, plunger-removable subpopulation enriched for B lymphocytes . The non-adherent T-cell subpopulation responded strongly to the T-cell mitogen PHA, but was unresponsive to both the peptidoglycan and cell walls from A . viscosus . In contrast, the weakly-adherent enriched B-cell subpopulation was less responsive to PHA, but was strongly stimulated by A . viscosus peptidoglycan and cell walls . These results indicate that peptidoglycan and cell walls from A . viscosus are B-cell mitogens. Prikl Biokhim Mikrobiol, 1985 Jan-Feb, 21(1), 114 - 21 {Bioluminescent method of determining picomolar amounts of nicotinamide-adenine dinucleotide using an immobilized extract of the luminescent bacterium Beneckea harveyi}; Lebedeva OV et al.; The luminous bacteria Beneckea Harveyi were immobilized on BrCN-sepharose and cellulose films activated with cyanuric chloride . Preparations with high luciferase and FMN-reductase activities were obtained, which showed no background luminescence without NADH being added . The storage conditions for the preparations obtained were optimized, and their kinetic parameters and thermostability were studied . Standard curves for NADH determining within the concentration range 1 nM-1 microM were plotted with the detection level of 1 picomol NADH . The preparations are very promising for bioluminescent assay due to their high activity, simple production, high stability during storage and a possibility for the repeated use. Arch Oral Biol, 1985, 30(11-12), 855 - 8 The comparative cariogenicity and plaque-forming ability in vivo of four species of the bacterium Actinomyces in gnotobiotic rats; Shakespeare AP et al.; Gnotobiotic WAG/RIJ rats on a high sucrose diet were monoinfected with 12 strains of Actinomyces spp . Moderate levels of caries were induced by a single strain of Actinomyces naeslundii and low levels by two strains of Actinomyces viscosus, three strains of A . naeslundii and one strain of Actinomyces israelii . No caries was induced by single strains of A . viscosus and A . israelii or by three Actinomyces odontolyticus strains . Only fissure caries was observed . Scanning electron microscopy showed that all strains colonized the fissures and most colonized the lingual surface of the teeth, but to a limited extent . Production of abundant and dense plaque was not always accompanied by caries. Arch Oral Biol, 1985, 30(9), 661 - 6 The mitogenicity for murine splenocytes of specific surface components of the oral periodontopathic bacterium, Actinomyces viscosus; Halfpap LM et al.; Many characterized fractions obtained from A . viscosus were examined to identify the macromolecules responsible for mitogenicity for lymphocytes . Spleen-cell suspensions of CBA/J mice were cultured with 50-200 micrograms dry weight of A . viscosus strains T14V and T14AV cellular components . Lipopolysaccharide (LPS) (Escherichia coli) was used as a positive control . Mechanical disruption with a French pressure cell or sonication produced preparations with a stimulation of 69,082 and 45,183 counts above background (CAB), respectively . Mitogenic activity was also present in the culture supernatant (38,000 CAB) . Other poorly mitogenic fractions included the peptidoglycan, cell-wall fractions, muramidase digests of cell walls, and the microcapsule extracted from whole cells with 0.5 M MgNO3 . The results suggest that mitogenic activity is not associated with the isolated cell-wall structure . The activity was released from the cell surface by physical shearing forces, as well as released into the medium growth. Recent Results Cancer Res, 1985, 98, 135 - 41 Effect of intensive adjuvant chemotherapy on wound healing in 69 patients with osteogenic sarcomas of the lower extremities; Bertermann O et al.; Reported surgical adjuvant trials in humans have resulted in little clinically significant impairment of wound healing . Such trials are often carried out with subtherapeutic doses or with the drugs administered relatively late in the wound healing process . It is the objective of our study to investigate the effect of intensive pre- and postoperative chemotherapy (BCD, ADR, HD-MTX) on wound healing in patients with osteogenic sarcomas . Wound healing was defined in our study as lack of infection . In a series of 110 patients with osteogenic sarcomas we analyzed the data of 69 patients with lower extremity lesions: of these, 54 patients had distal femur lesions and 15 had upper tibia and fibula lesions . All the patients underwent en-bloc resections and insertion of a prosthetic device . Pre- and postoperative antibiotics were given routinely . In 80% of our patients (55/69) an uneventful postoperative course was recorded with respect to wound healing . None of these required a secondary operative procedure . Debridement of the wound or debridement of the wound followed by skin grafting had to be performed in 14% (10/69) of the patients . Most of the amputations were performed early in this series of cases . After secondary surgery wound healing was uneventful in most of the patients . No patient died as a consequence of wound infection . Mixed bacterial infections were found in 13/14 patients . No single specific bacterium could be identified . One patient developed a fungal infection (aspergillosis) . Eight infections were secondary to skin necrosis . In this series we later found the serious effects of the skin necrosis and slough could be reduced by early intervention with a muscle pedicle flap.(ABSTRACT TRUNCATED AT 250 WORDS) Gene, 1985, 38(1-3), 233 - 7 A shuttle vector plasmid for studying carcinogen-induced point mutations in mammalian cells; Seidman MM et al.; We have constructed a shuttle vector plasmid for studying mutagenesis in mammalian cells . The plasmid replicates in cell lines permissive for SV40 virus as well as in the bacterium Escherichia coli and carries a bacterial suppressor tRNA gene (supF) that can serve as a mutagenesis marker . The plasmid replicates as efficiently as SV40 virus in African Green Monkey kidney CV1 cells, indicating that all traces of the inhibitory sequences normally found in pBR322 and its derivatives have been removed . The design of the plasmid and the small size of the mutagenesis target gene decrease the probability of recovering spontaneous deletion mutations that have been shown to occur at high frequency during passage in mammalian cells . The frequency of spontaneous-mutant plasmids recovered after passage in CV1 cells is substantially lower than with other vectors described previously . When the plasmid DNA is treated with UV radiation before passage in CV1 cells, mutants are observed at a frequency about 20-fold above the spontaneous background. Gene, 1985, 34(2-3), 367 - 70 Cloning of a multicopy plasmid from the actinorhizad nitrogen-fixing bacterium Frankia sp . and determination of its restriction map; Normand P et al.; An 8.3-kb multicopy plasmid, pFQ31, from the nitrogen-fixing Frankia sp . strain ArI3, was cloned into Escherichia coli plasmid vectors and analysed physically . pFQ31 has no detectable sequence homology with another Frankia plasmid, pFQ32, which is present in the same host . Derivatives of pFQ31 with an antibotic resistance marker were introduced into Streptomyces lividans, which is taxonomically related to Frankia, but no stable replication could be achieved. Acta Microbiol Pol, 1985, 34(1), 5 - 14 Autolysis of Caulobacter crescentus grown in the presence of glycine; Markiewicz Z et al.; Caulobacter crescentus was grown in complex medium supplemented with low (0.05%) concentration of glycine, a component of the murein peptide side chains of this bacterium . Murein synthesized in the presence of glycine was poorly crosslinked and the rate of its synthesis was slowed down compared to the control cells . The glycine-grown cells were considerably more sensitive to the chelating agent EDTA and Tris buffer than the control cells and also lysed faster when incubated with beta-lactam antibiotics . No changes in phospholipid composition in the presence of glycine were observed and the outer membrane protein composition of the glycine-grown cells was altered only in the amount of 130 000 protein which forms the surface array of C . crescentus . The effects of glycine can thus be tentatively put down to the reduced crosslinkage of murein synthesized in its presence. Acta Microbiol Pol, 1985, 34(2), 121 - 9 Murein hydrolases of Caulobacter crescentus; Markiewicz Z; Caulobacter crescentus was found to exhibit a similar autolytic response to a variety of factors affecting the structure of the cell envelope and interfering with murein synthesis as several other species of bacteria . Autolysis was accompanied by the hydrolysis of murein with the release of soluble degradation products . Several murein hydrolases with different bond specificity were found and except for the absence of DD-carboxypeptidase and LD-carboxypeptidase activities the make-up of these enzymes resembled that of the well studied bacterium Escherichia coli. Biochem Biophys Res Commun, 1984 Dec 28, 125(3), 1025 - 32 A cytoplasmic nickel-iron hydrogenase with high specific activity from Desulfovibrio multispirans sp . N., a new species of sulfate reducing bacterium; Czechowski MH et al.; A hydrogenase from a new species of sulfate reducing bacterium has been isolated and characterized . In contrast to other hydrogenases isolated from Desulfovibrio, this enzyme is found in the cytoplasmic fraction rather than in the periplasm . The specific activity of the enzyme, as measured in the hydrogen evolution assay, is twice as high as the specific activity of the hydrogenase from D . gigas . It also differentiates itself from the periplasmic Desulfovibrio hydrogenases by being more active in the hydrogen evolution rather than in the hydrogen uptake assay . The enzyme was shown to contain 0.9 atoms of nickel, 11 atoms of iron and 10 atoms of labile sulfide per mole of enzyme . It exhibits an unusually low intensity of the g = 2.31 nickel EPR signal in the isolated enzyme but shows a normal intensity for the g = 2.19 nickel EPR signal when reduced under hydrogen. J Biol Chem, 1984 Dec 25, 259(24), 15502 - 8 Purification and properties of the activating enzyme for iron protein of nitrogenase from the photosynthetic bacterium Rhodospirillum rubrum; Saari LL et al.; The oxygen-labile, activating enzyme for iron protein from the photosynthetic bacterium, Rhodospirillum rubrum, was purified 11,800-fold using a combination of chromatophore washing, DE52-cellulose chromatography, hydroxylapatite chromatography, reactive red-120 cross-linked agarose chromatography, reactive red-120 cross-linked agarose chromatography, and Sephadex G-75 gel filtration . Activating enzyme appeared homogeneous on silver-stained sodium dodecyl sulfate-polyacrylamide gels, and the staining intensity of the activating-enzyme band was correlated with the activating-enzyme activity observed in in vitro assays . Either formaldehyde fixation or higher acrylamide concentration was required to accurately assess the purity of activating enzyme on silver-stained gels . Activating enzyme was stable for 30 days at 4 degrees C . Dithiothreitol was a necessary component for the stability of partially purified activating enzyme . NaCl inhibited the coupled assay for activating enzyme . The pI of activating enzyme was determined to be 6.5 . Activating enzyme is composed of a minimum of 336 amino acids and a minimum calculated Mr is 32,032 . The Mr of activating enzyme was estimated to be 21,700 by analytical gel filtration and 32,800 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . An absorption maximum at 280 nm was observed for the activating enzyme. J Biol Chem, 1984 Dec 10, 259(23), 14458 - 62 Molecular structure of trimethylamine dehydrogenase from the bacterium W3A1 at 6.0-A resolution; Lim LW et al.; An electron density map of trimethylamine dehydrogenase has been calculated at 6.0-A resolution . Protein phases were based on two isomorphous mercury derivatives with similar binding properties, and on anomalous scattering measurements . The map has been averaged about the noncrystallographic 2-fold axis, plotted on transparent sheets and used to construct a wooden model . The elipsoidal dimer has a large inter-subunit interface . Each subunit appears to contain three closely associated domains with the iron-sulfur cluster located between two of them . The map suggests an alpha/beta-structure for two of the domains and a large helix content for the third. FEBS Lett, 1984 Dec 10, 178(2), 343 - 50 Two distinct quinone-modulated modes of antimycin-sensitive cytochrome b reduction in the cytochrome bc1 complex; Robertson DE et al.; Reduction of cytochrome b-560 (analogous to cyt b-562 of mitochondria) via an antimycin-sensitive route has been revealed in chromatophores of the photosynthetic bacterium, Rhodopseudomonas sphaeroides Ga . Indeed, the results suggest that two reductive mechanisms can be operative . One is consistent with the idea that the quinol generated at the reaction center QB site enters the Q pool and, via the Qc site, equilibrates with cytochrome b-560 . The other reductive mode circumvents redox equilibrium with the pool; we consider that this could result from a direct encounter of the reaction center with the bc1 complex perhaps involving a direct QB-Qc site interaction . This latter reaction is suppressed by occupancy of the Qc site, not only by antimycin but by ubiquinol and ubiquinone. J Mol Biol, 1984 Dec 5, 180(2), 385 - 98 X-ray structure analysis of a membrane protein complex . Electron density map at 3 A resolution and a model of the chromophores of the photosynthetic reaction center from Rhodopseudomonas viridis; Deisenhofer J et al.; X-ray analysis of three-dimensional crystals of the photosynthetic reaction center from the purple bacterium Rhodopseudomonas viridis led to an electron density distribution at 3 A resolution calculated with phases from multiple isomorphous replacement . The protein subunits of the complex were identified . An atomic model of the prosthetic groups of the reaction center complex (4 bacteriochlorophyll b, 2 bacteriopheophytin b . 1 non-heme iron, 1 menaquinone, 4 heme groups) was built . The arrangement of the ring systems of the bacteriochlorophyll b and bacteriopheophytin b molecules shows a local 2-fold rotation symmetry; two bacteriochlorophyll b form a closely associated, non-covalently linked dimer ("special pair") . A different local 2-fold symmetry axis is observed for the heme groups of the cytochrome part. Biophys J, 1984 Dec, 46(6), 781 - 6 Dispersal of motile bacteria from a plane layer; Cridland JV et al.; The dispersal of an initially well-defined concentration of the motile bacterium Escherichia coli was measured under nonchemotactic conditions . The distribution of bacteria along a glass observation cell was measured by recording the intensity of light scattered by the organisms . For comparison, the diffusion of fluorescein was also measured by determining the distribution of fluorescence throughout the observation cell . The dispersal of bacteria from a plane layer, under nonchemotactic conditions, can be adequately described by the Gaussian solution of the diffusion equation. Can J Microbiol, 1984 Dec, 30(12), 1467 - 76 Oxygen toxicity in Treponema pallidum: deoxyribonucleic acid single-stranded breakage induced by low doses of hydrogen peroxide; Steiner BM et al.; The effect of hydrogen peroxide on Treponema pallidum was investigated . The in vitro loss of virulence (as measured by rabbit inoculation) of T . pallidum was accelerated by as low as 100 microM hydrogen peroxide in the complex maintenance medium used . Higher doses led to rapidly accelerated death with 500 microM hydrogen peroxide causing sterilization of the medium within 3 to 4 h . Since hydrogen peroxide is known to cause single-stranded breaks in DNA, the effect of hydrogen peroxide on the treponemal genome was examined . Extensive breakage was caused by 100 microM hydrogen peroxide as determined on alkaline sucrose gradients . A limit was reached at 250 microM and above . Single-stranded breaks could be demonstrated as early as 5-10 min after exposure to hydrogen peroxide when the treponemes were exposed to 250 microM hydrogen peroxide; accelerated death was evident by 2 h past exposure demonstrating that DNA breakage was preceding death . Treponemal death caused by penicillin did not result in DNA breakage . The repair-proficient bacterium Escherichia coli K-12 was compared with T . pallidum . It required 10-100 times more hydrogen peroxide to cause various levels of breakage . Escherichia coli K-12 rapidly repaired DNA breakage once hydrogen peroxide was removed by addition of catalase . Treponema pallidum, in comparison, showed little or no repair in vitro . Addition of catalase or dithiothreitol to the medium protected against all but a low level of breakage; this may reflect on the ability of catalase and reducing agents to protect T . pallidum against oxygen toxicity in vitro. J Bacteriol, 1984 Dec, 160(3), 971 - 5 Isolation of a recombination-deficient mutant of Rhodopseudomonas capsulata; Genthner FJ et al.; To facilitate genetic analysis in the purple photosynthetic bacterium Rhodopseudomonas capsulata, a recombination-deficient derivative was sought . A UV irradiation-sensitive mutant (FG106F) was isolated after mutagenesis, and two procedures were used to determine the recombinational capacity of the mutant . First, recombinants were not detected after transduction of this derivative by the phage-like vector gene transfer agent . Second, an R-prime plasmid containing appropriately marked genes for photosynthesis was introduced by conjugation, and again no recombinants were observed . Additional phenotypes displayed by the mutant that are characteristic of a defect in recombination were an increased sensitivity to DNA-damaging antibiotics and a tendency to filament. J Bacteriol, 1984 Dec, 160(3), 1137 - 45 Cloning of the major protein of the Caulobacter crescentus periodic surface layer: detection and characterization of the cloned peptide by protein expression assays; Smit J et al.; A precisely ordered crystalline array is found on the surface of the bacterium Caulobacter crescentus CB15 . Using an immunological assay, we identified recombinant bacteriophage clones expressing the predominant protein of this structure from a lambda 1059 library of C . crescentus CB15 DNA . A single 4.4-kilobase HindIII fragment encoded a polypeptide whose antigenic determinants, molecular weight, and peculiar solubilization properties were identical with those of the authentic predominant polypeptide (130K) of the surface array . The 130K protein was produced as a discrete product as a result of gene transcription initiated from a lambda promoter; several experiments suggested that the Caulobacter promoter for this gene is not efficiently recognized by the Escherichia coli transcription machinery . Genomic Southern analysis revealed a single copy of the 130K protein gene per genome . The 130K protein gene was hybridized with DNA of two closely related laboratory strains of C . crescentus which have lost their ability to produce a surface array . One of these strains, CB2, possesses an homologous copy of the 130K gene, whereas DNA from the other strain, CB13B1a, showed a lesser degree of hybridization to the 130K gene probe; genomic fragments which did hybridize were of different sizes in CB13 as compared with those of CB15 . These findings are discussed in relation to studies of the surface array function and its role in cellular morphogenesis in this stalk-forming bacterium. Sci Total Environ, 1984 Nov, 39(3), 237 - 49 Incidence of Legionella organisms in selected Ontario (Canada) cities; Dutka BJ et al.; The distribution of Legionella pneumophila in water inside buildings was examined by means of culture methods . Cooling tower sumps and condenser valves harboured the organism at the highest frequency and in the highest concentrations . The bacterium was also frequently isolated from potable water systems, including hot and cold mixed taps, drinking water fountains and showers . When water quality parameters were examined, only elevated pH, total particulate nitrogen and alkalinity were correlated with the occurrence of L . pneumophila . Survival of the organism in water was increased at slightly basic pH and lower temperatures . The proliferation of the organism in water within buildings is probably due to a number of interrelated environmental factors that influence its survival and growth. Arch Microbiol, 1984 Nov, 139(4), 319 - 25 Construction of a gene bank of Rhodopseudomonas capsulata using a broad host range DNA cloning system; Klug G et al.; A gene bank of the phototrophic bacterium Rhodopseudomonas capsulata was constructed using the binary plasmid system pRK290/pRK2013 . Fragments of about 20 kb of chromosomal DNA of R . capsulata strain 37b4 were inserted into the cloning vector pRK290 . The hybrid plasmids of the gene bank, maintained in Escherichia coli HB101 were transferred by conjugation to R . capsulata strains defective in the photosynthetic apparatus with frequencies of 5 X 10(-4) to 5 X 10(-2) . Phototrophically growing transconjugants occurred with frequencies of 5 X 10(-7) to 5 X 10(-6) . Recombination between the hybrid plasmids and the R . capsulata chromosome was shown . The hybrid plasmid pRCF1002, carrying a 25 kb insert of R . capsulata wild type DNA, was isolated from one E . coli clone of the gene bank . It reconstituted some bacteriochlorophyll- and photosynthetic negative mutants to phototrophic growth. Chem Biol Interact, 1984 Nov, 52(1), 79 - 92 Irreversible binding of the chloropromazine radical cation and of photoactivated chlorpromazine to biological macromolecules; de Mol NJ et al.; The irreversible binding of chlorpromazine radical cation (CPZ+.) and photoactivated chlorpromazine (CPZ) to calf thymus DNA in vitro and bacterial macromolecules in intact bacterium cells was investigated . CPZ+ . may be formed in vivo metabolically and photochemically . CPZ+ . and photo-activated CPZ bind covalently to double- and single-strand DNA . The conformation of the DNA appeared to be important for the CPZ+ . reactivity: though CPZ+ . is less stabilized by complex formation with single-strand DNA, the reaction rate and the binding capacity of DNA-complexed CPZ+ . with single-strand DNA is larger than with double-strand DNA . Photoactivated CPZ binds considerably to proteins, DNA and RNA in the intact bacterium cells . In spite of the relatively short lifetime of CPZ+ . in the presence of the cells CPZ+ . also binds irreversibly to bacterial DNA, RNA and proteins . The consequences of covalent binding for the cytotoxicity and genotoxicity of CPZ+ . and photoactivated CPZ and the possible role for CPZ+ . as an intermediate in the photobinding of CPZ is discussed. Genetika, 1984 Nov, 20(11), 1783 - 91 {Inheritance of hybrid plasmid pAS*-21 by cells of the purple phototropic nitrogen-fixing bacterium Rhodopseudomonas sphaeroides}; Dubeikovskii AN et al.; Clones of purple nitrogenfixing prototrophic bacterium Rhodopseudomonas sphaeroides carrying an incertion into the chromosome of the hybrid pAS8-121 (RP4-ColE1 (repA::Tn7} plasmid were analysed . It is revealed that plasmid integration could be due to both Tn7 and other migrating elements (IS8 and possibly, to resident migrating elements of purple bacteria) . The plasmid pAS 8-121 can be autonomously transferred from the cointegrate state into Escherichia coli K-12; the plasmid is not inherited autonomously in cells of the purple bacterium . In E . coli cells R' derivatives of the plasmid carrying R . sphaeroides chromosomal fragments may be formed . The R' plasmids with fragments of plasmid DNA substituted for chromosomal material of R . sphaeroides were selected in E . coli K-12 cells among deleted (Kms) derivatives of pAS8-121. J Mol Biol, 1984 Oct 25, 179(2), 185 - 214 Rhodopseudomonas blastica atp operon . Nucleotide sequence and transcription; Tybulewicz VL et al.; The nucleotide sequence has been determined of a 12,368 base-pair region of DNA cloned from the non-sulphur photosynthetic bacterium Rhodopseudomonas blastica . It contains a cluster of six genes of which five encode the subunits of F1-ATPase; the sixth codes for an unknown protein . The genes are arranged in the same order as in the Escherichia coli unc operon, except that the unknown gene is placed between those for gamma and beta subunits . Neither the genes for F0 subunits, nor a homologue of the E . coli uncI gene is associated with this locus . The six genes are transcribed from a single promoter and we have designated this region the R . blastica atp operon . The two distal genes, beta and epsilon, may also be transcribed from a second promoter . Initiation and termination points for transcription have been identified by primer extensions and S1 nuclease mapping experiments . Signals involved in initiation of translation (Shine and Dalgarno sequences) and termination of transcription in the photosynthetic bacterium resemble those in E . coli . However, no common features can be identified in these two bacteria between 5' regions adjacent to sites of initiation of transcription . The sequence also contains a gene that encodes a protein homologous to discoidin, a cell surface lectin of Dictyostelium discoideum thought to be involved in cell--cell aggregation . Seven other reading frames have not been identified. Biochim Biophys Acta, 1984 Oct 17, 777(1), 41 - 55 Cell-cycle-specific biosynthesis of the photosynthetic membrane of Rhodopseudomonas sphaeroides . Structural implications; Yen GS et al.; Structural changes association with the intracytoplasmic membrane during the cell cycle of the photosynthetic bacterium Rhodopseudomonas sphaeroides have been studied by freeze-fracture electron microscopy . The isolated intracytoplasmic membrane vesicles, chromatophores, were fused in order to obtain large fracture faces, allowing more precise measurements and statistical analysis of both intramembrane particle density and size determinations . The intramembrane particle density of the protoplasmic face (PF) of the intracytoplasmic membrane, (from 4970 to 8290/micrometers 2), was shown to be a linear function of the protein/phospholipid ratio (from 2.5 to 5.1, w/w) of the intracytoplasmic membrane . Under constant light intensity, both the average particle size and particle size distribution remained unchanged during the cell cycle . These results provide the structural basis for the earlier reported cell-cycle-specific variations in both protein/phospholipid ratio and alternation in phospholipid structure of the intracytoplasmic membrane of R . sphaeroides during photosynthetic growth . The average particle diameter in the PF face of the intracytoplasmic membrane was 8.25, 9.08 and 9.75 nm at incident light intensities of 4000, 500 and 30 ft X cd, respectively . When chromatophores were fused with small, unilamellar liposomes, the intramembrane particle density decreased as input liposome phospholipid increased, whereas the particle size remained constant and particle distribution became random. J Mol Biol, 1984 Oct 15, 179(1), 151 - 5 Crystallization and preliminary x-ray diffraction study of the 3-Fe ferredoxin II from the bacterium Desulfovibrio gigas; Sieker LC et al.; The 3-Fe ferredoxin (FdII) from the bacterium Desulfovibrio gigas has been crystallized at pH 5.0 and 23 degrees C in two different crystal forms . One form is monoclinic, space group C2, with unit cell parameters a = 40.78 A, b = 44.98 A, c = 26.47 A, beta = 104.6 degrees, and one monomer of the FdII tetramer per asymmetric unit . The molecule can be either the monomer of molecular weight 6400 or a dimer of twice this molecular weight with 2-fold symmetry coincident with the 2-fold axis of the crystal . The other crystal form is orthorhombic, space group P2(1)2(1)2 and unit cell parameters a = 109.5 A, b = 37.0 A, c = 30.5 A . The asymmetric unit of this crystal contains two monomers of FdII . The orthorhombic crystal has not been reproduced since the initial crystallization. Gan To Kagaku Ryoho, 1984 Oct, 11(10), 2227 - 35 {Preparation of repeatedly and effectively usable L-asparaginase by a chemical modification}; Nishimura H et al.; The anti-tumor activity of L-asparaginase (EC . 3.5.1.1.) has been conclusively demonstrated . Its therapeutic application, however, has been hampered by its short clearance time in the circulation and high immunogenicity . In order to solve these problems, polyethylene glycol, a linear synthetic and non-immunogenic polymer, was introduced covalently into the amino groups in the enzyme molecule . Monomethoxypolyethylene glycol with molecular weight of 5000 was activated with cyanuric chloride to obtain activated PEGs {2, 4-bis (O-methoxypolyethyleneglycol)-6-chloro-s-triazine} . L-Asparaginase lost its immunoreactivity against its antibodies after the modification of 52 out of 92 amino groups in the molecule . This modified asparaginase retained 11% of its enzymic activity under the physiological condition . When the modified asparaginase was administered in rodents, it diminished the serum asparagine level and its enzymic activity persisted in the circulation 10 to 20 times longer than that of native enzyme . Furthermore, repeated injections of modified asparaginase did not induce any significant anti-asparaginase antibody production . Modified asparaginase showed a superior anti-tumor activity to the native counterpart irrespective of the presence of anti-asparaginase antibodies, when it was tested with murine Gardner lymphoma . This chemical modification should make it possible for the first time to repeatedly and effectively use L-asparaginase obtained from a bacterium. Environ Res, 1984 Oct, 35(1), 228 - 36 Growth rate of the bacterium Sphaerotilus natans in lead-containing medium and the effect of calcium ions; Medon P et al.; Growth rate of bacteria Sphaerotilus natans in lead-containing medium (1 X 10(-4) M Pb2+) and bioaccumulation of lead in the bacterial cells were investigated together with the effect of calcium ions on these processes . A rapid increase in biomass was observed in the presence of calcium and lead whereas calcium alone had no visible effect on the bacterial growth . The increase in biomass in the presence of lead and calcium was accompanied by a pronounced drop in lead content in the bacterial cells . This suggests that calcium ions protect the bacterial cells against lead poisoning. J Submicrosc Cytol, 1984 Oct, 16(4), 619 - 23 Photosynthetic reaction centers in artificial membranes: estimating protein dimensions by freeze-fracture and freeze-etching; Miller KR et al.; Because estimates of size and shape for membrane proteins are difficult to obtain directly, many workers have incorporated purified proteins into artificial lipid bilayers . Freeze-fracturing has then been used to provide a measure of the approximate size and shape of the membrane protein . We have formed reconstituted membranes containing the photosynthetic reaction center of Rhodopseudomonas viridis, a photosynthetic bacterium . The size and shape of this reaction center is accurately known from studies of negatively stained crystals of the protein to be approximately 4.5 X 6.0 nm . Freeze-fracture images of the reaction center in phosphatidyl choline liposomes show particles formed after reconstitution with an average diameter of 12.3 nm, much larger than the actual size of the protein . Deep-etched images of the surfaces of the liposomes, in which each individual reaction center complex can be seen clearly, show why the diameter of the protein is exaggerated in freeze-fracture . The individual reaction centers tend to cluster into large groups, allowing several individual reaction centers to be visualized as a single (much larger) particle in freeze-fracture . Freeze-fracturing, although a valuable tool in the analysis of membrane structure in natural and artificial membranes, must be used with caution in the estimation of molecular sizes and shapes. J Appl Bacteriol, 1984 Oct, 57(2), 279 - 90 A general-purpose system for characterizing medically important bacteria to genus level; Feltham RK et al.; A computer program and accompanying data matrix have been prepared for bacteria of medical interest, to assist the assignment of an unidentified bacterium to the most likely genus . The results on a set of relatively simple tests are entered . The program prints the more likely genera, followed by a list of diagnostic tables in Cowan & Steel (1974) and Buchanan & Gibbons (1974) . Where available, identification matrices for further computer-assisted study, are presented . This program may be of particular help in laboratories where a wide range of bacteria have to be identified. Arch Biochem Biophys, 1984 Oct, 234(1), 73 - 81 Comparison of the properties of two hydrogenases from the photosynthetic bacterium Chromatium vinosum; Serra JL et al.; Chromatium vinosum hydrogenases I and II were purified to specific activities of 9.6 and 28.0 units/mg protein, respectively . They have the same isoelectric point (pI = 4.1), and their visible spectra are typical of iron-sulfur proteins . Hydrogenase II in general was more stable than hydrogenase I . Both enzymes lost their activities slowly during storage in air, and this inactivation was more apparent in preparations of hydrogenase I . Bovine serum albumin helped to stabilize hydrogenase I against thermal and storage inactivation . The pH optima of H2-evolution activity of hydrogenases I and II were 7.4 and 5.4, respectively . Neither enzyme was able to evolve H2 from reduced ferredoxins as the sole electron carrier, but ferredoxins had an effect on the activity with methyl viologen as carrier to hydrogenase I . None of the natural compounds tested was able to serve as a physiological donor for H2 production . Hydrogenase I was more susceptible than hydrogenase II to inhibition by heavy metal ions and other enzyme inhibitors . Both enzymes were reversibly inhibited by CO with Ki values of 12 and 6 Torr for hydrogenase I and II, respectively . Hydrogenase I was more sensitive to denaturation by urea and guanidinium chloride while hydrogenase II was more susceptible to sodium dodecyl sulfate . Both enzymes were rapidly and irreversibly inactivated by dimethyl sulfoxide . Hydrogenase I evolved H2 from methyl viologen and ferredoxin photoreduced by chloroplasts . The enzymes differed in their iron and acid-labile sulfur contents. Biochem Biophys Res Commun, 1984 Sep 28, 123(3), 1234 - 9 Purification and properties of cytochrome b from photosynthetic bacterium Rhodopseudomonas sphaeroides R-26; Yu L et al.; Cytochrome b of R . sphaeroides R-26 has been purified from the isolated cytochrome b-c1 complex to homogeneity . The purification procedure involves Triton X-100 and urea solubilization, calcium phosphate column chromatography at different pH values, and ammonium sulfate fractionation . The purified protein contains 23 nmol heme per mg protein and has an apparent molecular weight of 43,000, as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis . The spectral characteristics of purified cytochrome b are similar to those of cytochrome b in the active cytochrome b-c1 complex but with a lower absorbance . The amino acid composition has been determined and compared with cytochrome b purified from other sources. J Biochem (Tokyo), 1984 Sep, 96(3), 691 - 9 Proton correlation NMR studies of metabolism in Rhodopseudomonas palustris; Imai Y et al.; A 1H correlation NMR study is reported, on the metabolism of a photosynthetic bacterium, Rhodopseudomonas palustris, in dark and light anaerobic conditions . Alkali treatment as well as sonication of the cells were employed to follow the process of accumulation and decomposition of poly-beta-hydroxybutyrate (PHB) which is the reserve material for the bacterium . It was shown that synthesis of PHB from trans-crotonate proceeds in the granules of the cells . It was also demonstrated that under anaerobic light conditions photometabolism and glycolysis generally compete with concomitant synthesis and decomposition of PHB, respectively, and that glycolysis gradually replaces photometabolism with aging of the cells . In contrast, glycolysis is always predominant in the dark and PHB is primarily used as the carbon source . It was observed that photo-induced transport of beta-hydroxybutyrate through the membrane occurs when photometabolism and glycolysis are equally active in the light . The implications of this observation are briefly discussed. Isr J Med Sci, 1984 Sep, 20(9), 836 - 9 Spiroplasmas and the transfer of genetic material by transformation and transfection; Bove JM et al.; Two plasmids, pMH1 with 7 kbp and pM41 with 8 kbp were purified from Spiroplasma citri strains MH and M4 respectively . On the basis of guanine + cytosine content and restriction enzyme mapping, the two plasmids are different . The linearized pMH1 plasmid was introduced into Escherichia coli plasmid vector pBR328 and could be cloned in E . coli . Using radioactive probes specific for each plasmid, we found that pM41 was present in three additional S . citri strains and in three other spiroplasmas not belonging to the S . citri species . pMH1 was found as a free 7-kbp plasmid only in the S . citri strain MH . However, the pMH1 probe hybridized strongly with high molecular weight DNA of several S . citri strains and strains of spiroplasmas other than S . citri . The major membrane protein of S . citri, spiralin, is strongly antigenic and rabbit antibodies against whole S . citri cells strongly react with spiralin . Thus, the enzyme-linked immunosorbent assay (ELISA) has been used to screen E . coli clones that were transformed with HindIII-generated S . citri DNA fragments inserted into the HindIII site of pBR328 . One E . coli transformant strongly reacted in ELISA with S . citri polyclonal antiserum . The same transformant also gave a positive reaction with monospecific antiserum against spiralin . These results demonstrate that a gene from S . citri, the spiralin gene, could be expressed in a bacterium . The isometric virus SV4, infecting honeybee spiroplasmas of Group I-2, was shown to possess circular single-stranded DNA of molecular weight 1.7 X 10(6) Da . Transfection of spiroplasma G1 with purified DNA of SV4 was achieved . These experiments open the way to the introduction of foreign genes into spiroplasmas. J Biochem (Tokyo), 1984 Sep, 96(3), 585 - 92 Ferredoxins from the photosynthetic purple non-sulfur bacterium Rhodopseudomonas palustris . Isolation and amino acid sequence of ferredoxin I; Minami Y et al.; Two ferredoxins, ferredoxins I and II, were prepared from Rhodopseudomonas palustris . They were separated on a Sephadex column after carboxymethylation and ferredoxin I, the major component, was subjected to an amino acid sequence study . The protein was composed of 63 amino acid residues and the sequence was as follows: (sequence; see text) . The molecular weight was calculated to be 6,718, excluding iron and sulfur atoms . The distribution of the nine cysteine residues was similar to but clearly distinct from those of ferredoxins of other photosynthetic bacteria . Comparison of this ferredoxin with those of other bacteria suggests that the photosynthetic bacteria evolved on separate lines . Ferredoxin II was also subjected to analyses of amino acid composition and terminal sequences, but no further study was possible due to the limited material . Although the composition was different from that of ferredoxin I, the terminal sequences were exactly the same as those of ferredoxin I. Pediatr Infect Dis, 1984 Sep-Oct, 3(5), 467 - 86 The concept of pertussis as a toxin-mediated disease; Pittman M; Pertussis (whooping cough), a two-stage process of disease (respiratory colonization and toxin-mediated disease) is caused by B . pertussis . The bacterium is unique . It is a pathogenic parasite with habitat only in human beings . Growth in the pathogenic form, both in in vitro and in vivo, requires conditions that permit the expression of pertussis toxin (PT) (also known as histamine-sensitizing factor, lymphocyte-leukocyte-promoting factor, islet-activating factor and pertussigen) . The expression of growth and PT appear to be genetically interrelated . For multiplication in vitro the medium must be free of substances, such as fatty acids, that inhibit the enzymatic action required for elaboration of PT . In vivo the bacteria are uniquely localized to the cilia of the respiratory epithelium where they multiply . In situ the bacteria inhibit natural defenses of the respiratory tract (cilial, phagocytic and other activities); they tend not to spread and do not invade the underlying tissue . The extent of the areas of colonization, directly related to the number of bacteria in the infecting inoculum, influences the amount of toxin elaborated and consequently the intensity of the clinical symptoms . Other factors that influence the clinical disease are the inordinate susceptibility of the infant and genetically controlled susceptibility . A specific role for PT in the initial establishment of the infection is not clear, but it seems definite that PT-specific immunity influences the clearance of colonization in about 4 to 5 weeks . The clinical symptoms become manifest when the bacteria are waning . This clearance is influenced by the synthesis of IgA antibodies and pertussis toxin antibodies that may act by inhibiting the "enzyme" required for growth or by another mechanism . The pathology of the disease is the result of altered cellular functions of toxin-sensitized cells, not by histologic damage . PT is composed of two functional components like other exotoxins that cause infectious disease (e.g . diphtheria, cholera) . Certain sites on one component enable PT to bind to specific receptors on tissue cells and enter the cell . The toxin ADP ribosylates a regulatory protein of the cytoplasmic membrane and thereby alters the function of the cell . Affected (sensitized) cells are insulin-secretory islets of the pancreas, lymphocytes and leukocytes, heart cells and others that have not been clearly identified, e.g . those that effect paroxysms and neurologic disturbances . The altered function of the cell in vitro is irreversible, and the restoration of the function of a particular tissue in vivo appears to be dependent on the renewal of the cells.(ABSTRACT TRUNCATED AT 400 WORDS) Proc Natl Acad Sci U S A, 1984 Sep, 81(18), 5816 - 20 Construction of Tn5 lac, a transposon that fuses lacZ expression to exogenous promoters, and its introduction into Myxococcus xanthus; Kroos L et al.; A promoterless trp-lac fusion fragment was inserted near one end of the bacterial transposon Tn5 in the correct orientation to fuse lacZ gene expression to promoters outside Tn5 . The resulting transposon, Tn5 lac, retains the kanamycin-resistance gene of Tn5 and transposes in Escherichia coli at 6% the frequency of Tn5 to many different sites in a bacteriophage lambda target . Expression of beta-galactosidase, the product of the lacZ gene, from Tn5 lac insertions in phage lambda depends both on insertion into a transcription unit in the correct orientation and on the regulation of the promoter of the transcription unit, verifying that by transposition Tn5 lac can fuse lacZ expression to outside promoters . An insertion of Tn5 lac in bacteriophage P1 was isolated and used to introduce Tn5 lac into Myxococcus xanthus, a bacterium that undergoes multicellular development . Stable kanamycin-resistant transductants are obtained that contain no P1 DNA sequences but have Tn5 lac inserted at different sites in the Myxococcus chromosome . Individual transductants express different levels of beta-galactosidase . A chromogenic substrate of beta-galactosidase, 5-bromo-4-chloro-3-indolyl beta-D-galactoside, is toxic in Myxococcus when cleaved in large amounts . In principle, Tn5 lac could be used to assay transcription in any bacterium in which Tn5 can transpose and beta-galactosidase can be measured. J Bacteriol, 1984 Aug, 159(2), 776 - 9 Changes in cell surface hydrophobicity of Myxococcus xanthus are correlated with sporulation-related events in the developmental program; Kupfer D et al.; Cell surface hydrophobicity was measured in the bacterium Myxococcus xanthus during vegetative growth, fruiting body formation, and glycerol-induced spore formation by the method of Rosenberg et al . (FEMS Microbiol . Lett . 9:29-33, 1980) . A significant decrease in cell surface hydrophobicity was observed 12 to 36 h after fruiting body formation and 60 to 120 min after glycerol-induced sporulation . The hydrophilic shift was correlated with the ability of the cells to sporulate but not with their ability to aggregate . Sucrose gradient purification removed the hydrophilic substance from the fruiting body spores but not from the glycerol-induced spores . The change in cell surface hydrophobicity in M . xanthus should be a useful developmental marker. Hastings Cent Rep . 1984 Aug;14(4):17. A cotton dust study unmasked; Levine C; KIE: The Dan River Company, citing news reports damaging to its image, has abandoned a proposed study to test a theory that byssinosis (brown lung disease) is caused by a bacterium growing in cotton rather than by inhalation of cotton dust . With state approval to exceed federal standards on cotton dust exposure, the company submitted the study to the Occupational Safety and Health Administration as a "variance," not as human subjects research . Levine contends that the proposal violated all major criteria of the federal regulations for protection of research subjects--scientific objectivity, balanced risks and benefits, and voluntary and informed consent . J Bacteriol, 1984 Aug, 159(2), 693 - 9 Carbon monoxide dehydrogenase from Rhodospirillum rubrum; Bonam D et al.; The carbon monoxide dehydrogenase from the photosynthetic bacterium Rhodospirillum rubrum was purified over 600-fold by DEAE-cellulose chromatography, heat treatment, hydroxylapatite chromatography, and preparative scale gel electrophoresis . In vitro, this enzyme catalyzed a two-electron oxidation of CO to form CO2 as the product . The reaction was dependent on the addition of an electron acceptor . The enzyme was oxygen labile, heat stable, and resistant to tryptic and chymotryptic digestion . Optimum in vitro activity occurred at pH 10.0 . A sensitive, hemoglobin-based assay for measuring dissolved CO levels is presented . The in vitro Km for CO was determined to be 110 microM . CO, through an unknown mechanism, stimulated hydrogen evolution in whole cells, suggesting the presence of a reversible hydrogenase in R . rubrum which is CO insensitive in vivo. Eur J Biochem, 1984 Jul 2, 142(1), 21 - 8 Purification, characterization and redox properties of hydrogenase from Methanosarcina barkeri (DSM 800); Fauque G et al.; A soluble hydrogenase from the methanogenic bacterium, Methanosarcina barkeri (DSM 800) has been purified to apparent electrophoretic homogeneity, with an overall 550-fold purification, a 45% yield and a final specific activity of 270 mumol H2 evolved min-1 (mg protein)-1 . The hydrogenase has a high molecular mass of approximately equal to 800 kDa and subunits with molecular masses of approximately equal to 60 kDa . The enzyme is stable to heating at 65 degrees C and to exposure to air at 4 degrees C in the oxidized state for periods up to a week . The overall stability of this enzyme is compared with other hydrogenase isolated from strict anaerobic sulfate-reducing bacteria . Ms . barkeri hydrogenase shows an absorption spectrum typical of a non-heme iron protein with maxima at 275 nm, 380 nm and 405 nm . A flavin component, identified as FMN or riboflavin was extracted under acidic conditions and quantified to approximately one flavin molecule per subunit . In addition to this component, 8-10 iron atoms and 0.6-0.8 nickel atom were also detected per subunit . The electron paramagnetic resonance (EPR) spectrum of the native enzyme shows a rhombic signal with g values at 2.24, 2.20 and approximately equal to 2.0 . probably due to nickel which is optimally measured at 40 K but still detectable at 77 K . In the reduced state, using dithionite or molecular hydrogen as reductants, at least two types of g = 1.94 EPR signals, due to iron-sulfur centers, could be detected and differentiated on the basis of power and temperature dependence . Center I has g values at 2.04, 1.90 and 1.86, while center II has g values at 2.08, 1.93 and 1.85 . When the hydrogenase is reduced by hydrogen or dithionite the rhombic EPR species disappears and is replaced by other EPR-active species with g values at 2.33, 2.23, 2.12, 2.09, 2.04 and 2.00 . These complex signals may represent different nickel species and are only observable at temperatures higher than 20 K . In the native preparation, at high temperatures (T greater than 35 K) or in partially reduced samples, a free radical due to the flavin moiety is observed . The EPR spectrum of reduced hydrogenase in 80% Me2SO presents an axial type of spectrum only detectable below 30 K. Biophys J, 1984 Jul, 46(1), 57 - 64 Magnetosome dynamics in magnetotactic bacteria; Ofer S et al.; Diffusive motions of the magnetosomes (enveloped Fe3O4 particles) in the magnetotactic bacterium Aquaspirillum magnetotacticum result in a very broad-line Mossbauer spectrum (T approximately 100 mm/s) above freezing temperatures . The line width increases with increasing temperature . The data are analyzed using a bounded diffusion model to yield the rotational and translational motions of the magnetosomes as well as the effective viscosity of the material surrounding the magnetosomes . The results are {theta 2} l/2 less than 1.5 degrees and {x2} 1/2 less than 8.4 A for the rotational and translational motions, respectively, implying that the particles are fixed in whole cells . The effective viscosity is 10 cP at 295 K and increases with decreasing temperature . Additional Fe3+ material in the cell is shown to be associated with the magnetosomes . Fe2+ material in the cell appears to be associated with the cell envelope. J Bacteriol, 1984 Jul, 159(1), 26 - 35 Isolation and characterization of nonspreading mutants of the gliding bacterium Cytophaga johnsonae; Chang LE et al.; Three approaches were taken to isolate a total of 153 nonspreading mutants derived from our laboratory strain of Cytophaga johnsonae, UW101, or from its auxotrophic derivative, UW10538 . Characterization of 109 of these mutants led to their placement in five general categories: (i) motile, nonspreading (MNS) mutants whose cells are motile to various degrees but whose colonies fail to spread on agar gels under any conditions of incubation; (ii) conditional nonspreading (CNS) mutants with motile cells whose colonies require more moisture to spread on agar gels than do those of wild-type cells; (iii) filamentous conditional motility (FCM) mutants whose cells grow as nonmotile filaments or as motile cells with wild-type morphology, depending on conditions of incubation; (iv) short, tumbling, nonspreading (STN) mutants with short cells that tumble constantly; and (v) truly nonmotile (TNM) mutants whose cells never move and whose colonies never spread under any conditions tested . All TNM mutants exhibited a remarkable pleiotropy not seen in the other four classes of mutants: all were resistant to 39 phages to which wild-type cells are sensitive, and all were unable to digest chitin, which is digested by wild-type cells . The correlation between ability to move and phage sensitivity was strengthened further by showing that 150 additional TNM mutants derived from UW101 and 43 TNM mutants derived from 29 independent isolates of C . johnsonae were resistant to all phages to which their parents were sensitive . Furthermore, motile revertants of TNM mutants became phage sensitive, and temperature-sensitive mutants were motile and phage sensitive at 25 degrees C and nonmotile and phage resistant at 32 degrees C . Evidence supports the conclusion that any mutation rendering cells truly nonmotile invariably alters cell surface-associated properties such as phage sensitivity and chitin digestion merely as a consequence of changing a moving cell surface to a static surface. J Bacteriol, 1984 Jul, 159(1), 222 - 7 Isolation of pigmentation mutants of the green filamentous photosynthetic bacterium Chloroflexus aurantiacus; Pierson BK et al.; Mutants deficient in the production of bacteriochlorophyll c (Bchl c) and one mutant lacking colored carotenoids were isolated from the filamentous gliding bacterium Chloroflexus aurantiacus . Mutagenesis was achieved by using UV radiation or N-methyl-N'-nitro-N-nitrosoguanidine . Several clones were isolated that were deficient in Bchl c synthesis . All reverted . One double mutant deficient both in Bchl c synthesis and in the synthesis of colored carotenoids under anaerobic conditions was isolated . Isolation of a revertant in Bchl c synthesis from this double mutant produced a mutant strain of Chloroflexus that grew photosynthetically under anaerobic conditions and lacked colored carotenoids . Analysis of pigment contents and growth rates of the mutants revealed a positive association between growth rate and content of Bchl c under light-limiting conditions. Microbiologica, 1984 Jul, 7(3), 263 - 6 A bald superfertile U.V.-resistant strain in Streptomyces coelicolor A3(2); Puglia AM et al.; The 'bald'(bld) mutants of filamentous bacterium Streptomyces coelicolor A3(2) are characterized by a lack of aerial myceliumand spores . A 'bald' mutant was isolated exhibiting unusual features . It forms slightly sculptured colonies producing a red-orange mycelial pigment, large amounts of agarase and methylenomycin A; it is also highly resistant to U.V . killing . The bld mutation (bld F1) never reverted to bld+ phenotype and was localized very closed to met A. Rev Infect Dis, 1984 Jul-Aug, 6(4), 579 - 86 Classics in infectious diseases . Toxic products of Bacterium enteritidis and of related micro-organisms . By Sara Elizabeth Branham . Journal of Infectious Diseases 1925; Biosynthesis of the sulfonolipid 2-amino-3-hydroxy-15-methylhexadecane-1-sulfonic acid in the gliding bacterium Cytophaga johnsonae; The biosynthesis of the sulfonolipid 2-amino-3-hydroxy-15-methylhexadecane-1-sulfonic acid (capnine) was studied by measuring the incorporation of possible precursors into the lipid by cells grown in the presence of precursors which were labeled with stable isotopes . Cells grown on yeast extract in the presence of DL-{3,3-2H2}serine contained 40.1 mol% of the protein-bound serine and 5.0 mol% of the protein-bound cysteine derived from the labeled serine . Cells grown in the presence of DL-{3,3-2H2}cystine acid contained 86.4 mol% of the molecules that had two deuteriums . These results are consistent with the possibility that biosynthesis of capnine occurs by the condensation of 13-methylmyristoyl-coenzyme A with cysteic acid, in a reaction analogous to the condensation of a palmitoyl-coenzyme A with serine to form 3-keto-sphinganine during the biosynthesis of sphingolipids. Infect Immun, 1984 Jul, 45(1), 101 - 6 Antigen from Francisella tularensis: nonidentity between determinants participating in cell-mediated and humoral reactions; Sandstrom G et al.; After tularemia vaccinations, most individuals respond with cell-mediated and humoral immunity as disclosed by the lymphocyte stimulation test and enzyme-linked immunosorbent assay (ELISA), respectively . There is, however, no correlation between the magnitudes of the two responses, and some individuals show one of the responses only . We now report that the two responses are directed towards different antigenic determinants of the bacterium . Ether-water extraction of the live vaccine strain of Francisella tularensis gave a high yield of material reacting in ELISA as well as in the lymphocyte stimulation test . However, the specificity of the extract was low insofar as it reacted not only with lymphocytes and antibodies of tularemia-vaccinated individuals but also to a fairly high extent with those of nonvaccinated individuals . By using the extract as a starting material, a preparatory procedure was developed, resulting in antigen of high specificity in the two tests . The antigen prepared was a high-molecular-weight, carbohydrate-protein complex . Proteinase K treatment of the antigen abolished the lymphocyte-stimulating activity but did not decrease ELISA activity at all . Periodate treatment, on the other hand, greatly reduced ELISA activity but did not decrease the lymphocyte-stimulating activity . Thus, determinants of F . tularensis responsible for immunospecific lymphocyte stimulation seem to reside in protein, whereas ELISA activity seems to be due mostly to carbohydrate determinants. Biochem Biophys Res Commun, 1984 Jun 29, 121(3), 755 - 61 Inhibition of photosy