J Med Microbiol, 1990 Aug, 32(4), 247 - 53 Detection of diphtheria toxin in culture supernates of Corynebacterium diphtheriae and C . ulcerans by immunoassay with monoclonal antibody; Hallas G et al.; Monoclonal antibody (MAb) to diphtheria toxin was produced in mouse hybridomas, and shown by ELISA to be of sub-class IgG1 . Hybridomas were inoculated into mice to produce ascitic fluid from which MAb was purified by caprylic acid . The MAb was shown by immunoblotting to be directed against the A fragment of the toxin and also against the intact toxin molecule . After conjugation with fluorescein isothiocyanate, it was used in an immunoassay to detect toxin in culture supernates of Corynebacterium diphtheriae and C . ulcerans . The assay correlated well with the Elek test and with virulence in guinea-pigs; but it gave occasional false positive results, probably by binding of MAb to defective toxin. Proc Natl Acad Sci U S A, 1990 Aug, 87(15), 5968 - 72 Molecular cloning and DNA sequence analysis of a diphtheria tox iron-dependent regulatory element (dtxR) from Corynebacterium diphtheriae; Boyd J et al.; Although the structural gene for diphtheria toxin, tox, is carried by a family of closely related corynebacteriophages, the regulation of tox expression is controlled, to a large extent, by its bacterial host Corynebacterium diphtheriae . Optimal yields of tox gene products are obtained only when iron becomes the growth-rate-limiting substrate . Previous studies suggest that regulation of tox expression is mediated through an iron-binding aporepressor . To facilitate molecular cloning of the tox regulatory element from genomic libraries of C . diphtheriae, we constructed a tox promoter/operator (toxPO)-lacZ transcriptional fusion in Escherichia coli strain DH5 alpha . We report the molecular cloning and nucleic acid sequence of a diphtheria tox iron-dependent regulatory element, dtxR, and demonstrate that expression of beta-galactosidase from the toxPO-lacZ fusion is regulated by dtxR-encoded protein in an iron-sensitive manner . In addition, we show that expression of the toxPO-lacZ fusion is not affected by the E . coli iron-regulatory protein Fur and that the dtxR protein does not inhibit expression of fur-regulated outer-membrane proteins. J Hosp Infect, 1990 Aug, 16(2), 123 - 32 Antigenic cross-reactivity among isolates of group JK corynebacteria; Moore MK et al.; Using rocket and rocket-line immunoelectrophoresis and immunoblotting it was demonstrated that a group of antibiotic-resistant bacteria, including several authenticated Corynebacterium jeikeium strains, shared many cross-reacting antigens . Only weak cross-reactivity was demonstrated with representatives of three other skin corynebacteria, C . bovis, C . hofmanii and C . minutissimum or with non-coryneforms . Differences within C . jeikeium are sufficient to permit the use of immunoblotting as an epidemiological tool. Eur J Immunol, 1990 Aug, 20(8), 1761 - 5 Corynebacterium parvum (Propionibacterium acnes): an inducer of tumor necrosis factor-alpha in human peripheral blood mononuclear cells and monocytes in vitro; Rossol S et al.; The present study investigates the potential capacity of the immunostimulant Corynebacterium parvum (C.p.) to induce tumor necrosis factor-alpha (TNF-alpha) in human peripheral blood mononuclear cells (PBMC) and blood monocytes (BMo) in vitro . Both at the mRNA and protein level, stimulation of PBMC and BMo upon C.p . induces TNF-alpha . Compared to the hitherto used TNF-alpha inducers in vitro such as Sendai virus, phytohemagglutinin or lipopolysaccharide the C.p . stimulus displayed a threefold stronger induction of TNF-alpha production (p less than 0.001) . Using C.p . as an inducer it was possible to demonstrate that TNF-alpha production is regulated by prostaglandin E2; preincubation of the cells with prostaglandin E2 resulted in a reduced C.p.-mediated TNF-alpha production (p less than 0.001) . Coincubation of interferon-gamma (IFN-gamma) together with C.p . led to an enhanced release of TNF-alpha, supporting the assumption that C.p . is a potent TNF-alpha inducer . The additive effect of IFN-gamma and TNF-alpha on the receptor level was demonstrated by addition of IFN-gamma antibodies to the PBMC cultures . Under these conditions TNF-alpha production, stimulated by C.p . and IFN-gamma, was decreased by 30%, compared to the production in assays supplemented with C.p . alone . From these data we conclude that C.p . is a new inducer of TNF-alpha in vitro and a useful tool to study TNF-alpha production of PBMC and BMo from either healthy donors or from patients. Cancer, 1990 Jul 15, 66(2), 220 - 7 Evaluation of the worth of corynebacterium parvum in conjunction with chemotherapy as adjuvant treatment for primary breast cancer . Eight-year results from the National Surgical Adjuvant Breast and Bowel Project B-10; Fisher B et al.; During the 1970s, information obtained from animal tumor models and from patients with a spectrum of solid tumors indicated the worth of a variety of immunostimulating agents . These findings provided a biological and clinical rationale for conducting randomized trials to evaluate the worth of those agents . Consequently, in May 1977 the National Surgical Adjuvant Breast and Bowel Project (NSABP) implemented a randomized trial to determine whether Corynebacterium parvum (C . parvum, CP) plus chemotherapy would be more effective than chemotherapy alone in prolonging the disease-free survival (DFS) and survival (S) of patients with primary operable breast cancer and positive axillary nodes . The results of that trial through 8 years of follow-up fail to indicate that treatment with CP used in conjunction with l-phenylalanine mustard (L-PAM) plus 5-fluorouracil (PF) results in a better DFS and S than that observed after chemotherapy alone . Use of the immunomodulator has instead resulted in a poorer, but not statistically significant, outcome . Despite adjustments made to account for any imbalance in distribution of prognostic factors between the two treatment groups and despite considering treatment compliance as a factor, the unfavorable outcome persisted . A high incidence of fever and chills was associated with the administration of CP . The administration of hydrocortisone before each CP treatment reduced the frequency of those and other systemic effects . The failure to demonstrate a benefit from CP is in keeping with the failure of other nonspecific stimulating agents to contribute to the creation of a new paradigm for the treatment of breast cancer. Infect Immun, 1990 Jul, 58(7), 2198 - 203 Vaccination of humans against cutaneous leishmaniasis: cellular and humoral immune responses; Nascimento E et al.; Brazilian army conscripts were vaccinated against American cutaneous leishmaniasis by using nonliving polyvalent promastigote Leish vaccine 5 or Leish vaccine 6 (vaccines with five or six Leishmania stocks) with or without Corynebacterium parvum . No statistically significant differences in lymphocyte stimulation indices were found between vaccinated groups with or without C . parvum, but lymphocyte stimulation indices of all vaccinees were significantly higher (P less than 0.001) than those of the placebo group . A correlation of 90% was found between positive skin test results and positive lymphocyte stimulation indices . Eight major antigens with estimated molecular masses of 13.5, 25, 40, 63, 73, 85, 97, and 160 kilodaltons were recognized by Leish vaccine 5 sera . Our finding also demonstrated the predominance of immunoglobulin M antibody in sera of vaccinated subjects and that a component of Leish vaccine 5, gp63, was immunogenic in humans both at the T-cell level and at the antibody level. Sb Lek, 1990 Jul, 92(6-7), 212 - 8 {New aspects of intolerance of the aqueous iodinated x-ray contrast medium, adipiodone--fatal interaction of adipiodone-Ultrabil with the immunomodulators Corynebacterium parvum (CP) and Shigella dysenteriae endotoxin (LPS)}; Klein O et al.; The authors assume a new type of interaction of the aqueous X-ray contrast substance type adipiodone (Ultrabil 50% Spofa) with immunomodulators of Corynebacterium parvum (CP) and the endotoxin of Shigella dysenteriae (LPS) . The fatal type of interaction developed regularly as a result of the concurrent i.v . administration of the X-ray contrast substance adipiodone with the endotoxin of Shigella dys . (LPS) to laboratory animals (mice) which had five days previously a single dose of Corynebacterium parvum (CP) . The effect of the interaction was evaluated using LD50 of adipiodone (Ultrabil Spofa) which in interaction with the immunomodulators of CP and LPS was identical as the dose used in diagnostic practice . The authors considered the possible development of "oxygen stress" as a consequence interacting processes and complement activation. Mycopathologia, 1990 Jul, 111(1), 9 - 15 Antimicrobial activity of naphthoquinones from fusaria; Baker RA et al.; Twenty-two naphthoquinone compounds isolated or derived synthetically from culture extracts of Fusarium solani and F . oxysporum were examined for antimicrobial activity . Fifteen exhibited antibiotic activity against Staphylococcus aureus, and 12 were active against Streptococcus pyogenes, but none were active at the highest rate of 128 micrograms/ml against Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Proteus vulgaris, Serratia marcescens, or Pseudomonas aeruginosa . Of 8 plant pathogenic bacteria tested against 11 naphthoquinones, Corynebacterium poinsettiae was inhibited by 6 compounds, and Pseudomonas viridiflava was weakly inhibited by one . Only one of a group of 6 fluorescent soil pseudomonads was inhibited by one naphthoquinone . Antifungal activity of 10 compounds against 8 fungal plant pathogens was limited to inhibition of Phytophthora parasitica by one naphthopyran. Nippon Saikingaku Zasshi, 1990 Jul, 45(4), 785 - 95 {Studies on immunity to Babesia gibsoni in dogs immunostimulation by Bordetella bronchiseptica}; Ohgitani T et al.; Four species of bacteria, Corynebacterium anaerobium 578, Actinobacillus pleuropneumoniae G-4, Mycobacterium bovis BCG, and Bordetella bronchiseptica A-2, were injected intravenously into mice (5 weeks old, ICR-SPF) . The clearance of carbon from the blood stream and the weights of the spleen and liver were determined as indicators of RES stimulation . Mouse footpad reaction was assessed as an indicator of delayed-type hypersensitivity to each species of bacteria . The immuno-stimulative activity of each species of bacteria against bovine serum albumin was monitored by passive hemagglutination assay and the macrophage migration-inhibition test in guinea pigs . Based on the results of the experiments described above, B . bronchiseptica was selected as an immunostimulator (Ims) for immunization trials of the hemo-protozoan parasite, Babesia gibsoni, with inactivated merozoites of B . gibsoni (BgK) . Twelve dogs, pointers about 6 months old, were divided into four groups of three dogs each . Group 1 dogs were initially injected with Ims, and later injected with BgK and Ims (BgK+Ims) after a 3-week interval . Group 2 and Group 3 dogs were injected twice, at a 3-week interval, with BgK+Ims and BgK, respectively, and Group 4 served as a control . As the results, the serum antibody titres of Group 1 and 2 were several times higher than that of Group 3, and the cell-mediated immunity to parasites was noticeably stimulated by immunization with BgK+Ims . The peak level of parasitemia following the challenge were over 10% for Group 4 and 4.5% for Group 3, while levels for Group 1 and 2 were 2.5% and less than 1%, respectively . No such major clinical signs of babesiosis as jaundice and anemia were observed in Group 1 or 2. Br J Clin Pract, 1990 Jul, 44(7), 285 - 7 Diphtheritic polyneuritis in an elderly woman: clinical and neurophysiological follow-up; Roche S et al.; A 71-year-old English lady initially presented with a bulbar paralysis and, six weeks later, developed a generalised sensori-motor neuropathy . Corynebacterium diphtheriae mitis was cultured from her throat swab . Despite a good clinical recovery at one month, nerve conduction velocity was at its lowest . As far as the authors are aware, this is one of the few cases of neurophysiological and clinical follow-up in a British subject with diphtheritic peripheral neuropathy . This case emphasises the importance of giving antitoxin early. Clin Microbiol Rev, 1990 Jul, 3(3), 227 - 46 Coryneform bacteria in infectious diseases: clinical and laboratory aspects; Coyle MB et al.; Coryneform isolates from clinical specimens frequently cannot be identified by either reference laboratories or research laboratories . Many of these organisms are skin flora that belong to a large number of taxonomic groups, only 40% of which are in the genus Corynebacterium . This review provides an update on clinical presentations, microbiological features, and pathogenic mechanisms of infections with nondiphtheria Corynebacterium species and other pleomorphic gram-positive rods . The early literature is also reviewed for a few coryneforms, especially those whose roles as pathogens are controversial . Recognition of newly emerging opportunistic coryneforms is dependent on sound identification schemes which cannot be developed until cell wall analyses and nucleic acid studies have defined the taxonomic groups and all of the reference strains within each taxon have been shown by molecular methods to be authentic members . Only then can reliable batteries of biochemical tests be selected for distinguishing each taxon. J Clin Microbiol, 1990 Jul, 28(7), 1586 - 90 Comparative evaluation of nonradiometric BACTEC and improved oxoid signal blood culture systems in a clinical laboratory; Daley C et al.; The BACTEC NR660 blood culture system, which uses infrared spectroscopy to detect carbon dioxide generated by bacterial growth, was compared with the new medium formulation of the Oxoid Signal system . Two trials were conducted: a comparative study of 88 organisms in simulated blood cultures and a clinical trial of 3,321 paired patient blood culture samples . Both trials showed that overall the BACTEC system performed better in the recovery of organisms . The Oxoid system was unable to detect by signal the growth of the majority of yeasts, nonfermentative gram-negative bacilli, Neisseria meningitidis, Nocardia spp., and Corynebacterium jeikeium . There were no significant differences in the yield of Staphylococcus spp., members of the family Enterobacteriaceae, Streptococcus spp., or anaerobic organisms . BACTEC detected growth more quickly than did the Oxoid system; 61% of the isolates were detected by BACTEC at 24 h, while 49% of the isolates were detected by Oxoid . The Oxoid system had a high proportion (58.5%) of false-positives, compared with 7.7% for the BACTEC system . Despite the new medium formulation of the Oxoid system, its performance is still not equivalent to that of the BACTEC system. Oral Surg Oral Med Oral Pathol, 1990 Jul, 70(1), 96 - 108 The risk of intentional dissolution of the smear layer after mechanical preparation of root canals; Gutierrez JH et al.; The root canals of 27 caries-free human cuspids were divided into 3 groups of 9 teeth each . Group I was mechanically prepared with reamers and files . Alternate irrigation with 5.25% NaOCl and 3% hydrogen peroxide was used between each instrument . The group II root canals were prepared with the use of alternate irrigation with the same concentrations of NaOCl and hydrogen peroxide and finally flushed with 17% EDTA and NaOCl solution . Group III was prepared mechanically with the use of sterile distilled water as an irrigant followed by a final flush with 17% EDTA solution . The root canals of all specimens were obturated with gutta-percha points and Tubliseal by means of the lateral condensation technique . After the root canals were obturated, samples of each of the three groups were immersed in separate flasks containing Streptococcus mutans, Staphylococcus epidermidis, and Corynebacterium hofmannii . All the microorganisms tested invaded the coronal dentin--where the access cavity had been sealed with Fynal--and/or the main foramen. Int Endod J, 1990 Jul, 23(4), 196 - 202 Penetration of dentine by three oral bacteria in vitro and their associated cytotoxicity; Meryon SD et al.; Three common oral bacteria, namely Streptococcus sanguis, Actinomyces viscosus and Corynebacterium spp . were studied with regard to their ability to penetrate etched and unetched dentine and for their effect on underlying cell cultures . The test organisms were grown in cylinders above dentine slices 100 and 500 microns thick for 72 hours . The slices were in contact with tissue culture medium covering a layer of fibroblasts . Penetration of 100 microns slices was most rapid with S . sanguis, followed by A . viscosus and Corynebacterium . The pattern was similar but slightly delayed when 500 microns slices were used, but in most cases penetration had occurred by 72 hours . The presence of a smear layer had no effect on the results obtained . Following penetration, cell destruction was most extensive with S . sanguis, the most cytotoxic organism, followed by Corynebacterium and A . viscosus . In the limited number of dishes where no penetration occurred there was little effect on cell numbers. Trans R Soc Trop Med Hyg, 1990 Jul-Aug, 84(4), 589 - 92 The bacteriology of brain abscess: a local experience in Malaysia; Puthucheary SD et al.; 31 cases of intracranial abscess seen over a period of 10 years showed a peak incidence in the second and third decades of life with a male preponderance . Tetralogy of Fallot and other congenital cyanotic heart diseases were the predominant associated factors (32%) . The commonest site of infection was the frontal lobe . Gram-stained smears of pus proved to be extremely useful . The majority of the organisms (82%) were either microaerophilic or anaerobic bacteria with Streptococcus milleri being the most frequent isolate . With the exception of Corynebacterium species, all isolates were susceptible to penicillin or chloramphenicol, most being susceptible to both. Kansenshogaku Zasshi, 1990 Jun, 64(6), 661 - 7 {A clinical study of bacteremia in urology}; Tokunaga S et al.; We investigated 32 patients with bacteremia that occurred in the Department of Urology, School of Medicine, Kanazawa University between April, 1983 and March, 1989 . This incidence represented 1.9% of the total number of inpatients . The study group comprised 29 males and 3 females, and their age varied from 25 to 82 years with a mean age of 61.7 years . Twenty-two (75%) of the 32 patients had urologic malignancies . The majority of patients were compromised hosts who had one or more (average, 3.8) factors that promoted bacteremia . Urinary tract infections existed in 26 (86.0%) patients before the bacteremic episode and urine cultures revealed a species identical to that simultaneously isolated from the blood in 19 (73.1%) of the 26 patients . Out of the 26 patients, there were 22 (84.6%) with complicated pyelonephritis and 22 (84.6%) with an indwelling urinary tract catheter . In blood cultures, the most common isolate was Staphylococcus epidermidis and gram-positive cocci were cultured at a rate of 43.9% which was higher than that (39.0%) of gram-negative rods . In contrast, in urine cultures, gram-negative rods were isolated predominantly . S . epidermidis and Corynebacterium spp . isolated less frequently in blood than in urine, indicated contaminants . However, Enterococcus spp . and Candida albicans were recognized as causative organisms of bacteremia via the urinary tract, because the urine culture demonstrated a species identical to that obtained from blood in these bacteremic patients . Antibiotic sensitivity tests demonstrated that isolates from blood tended to show tolerance to beta-lactam antibiotics, but had good sensitivity to aminoglycosides.(ABSTRACT TRUNCATED AT 250 WORDS) Am J Vet Res, 1990 Jun, 51(6), 874 - 7 Alterations in the phospholipid composition and morphology of ovine erythrocytes after intravenous inoculation of Corynebacterium pseudotuberculosis; Brogden KA et al.; Corynebacterium pseudotuberculosis produces a sphingomyelin-specific phospholipase D exotoxin that is a major determinant in the pathogenesis of caseous lymphadenitis . The effect of this exotoxin on erythrocytes was assessed during experimentally induced infection of sheep . Blood was drawn at timed intervals, and the phospholipid composition of erythrocytes was determined by use of high-performance liquid chromatographic analysis of membrane extracts . Erythrocyte morphology was determined by use of transmission electron microscopy . Significant (P less than or equal to 0.05) decreases in erythrocyte membrane sphingomyelin content and significant (P less than or equal to 0.05) increases in phosphatidylglycerol content were observed 30 minutes after IV inoculation of C pseudotuberculosis . The concentration of other phospholipids remained unchanged . Initially, spherostomatocytes were formed that later became pitted at the cell surface . These pits or invaginations appeared as numerous vacuoles at the periphery of thin-sectioned cells . Pitting became progressively worse, leading to an extensive scalloped cell surface . Alterations in the phospholipid composition and morphology of ovine erythrocytes may contribute to pathophysiologic findings in sheep with acute infection induced by C pseudotuberculosis. Genetika, 1990 Jun, 26(6), 990 - 9 {Cloning and expression of the gene for diphtheria toxin and its subunits in Escherichia coli}; Garaev MM et al.; The results of cloning Corynebacterium diphtheriae phi 984 tox gene and its A and B subunits in Escherichia coli are presented . Regulatory sequences of tox gene are capable to promote effective expression in E . coli cells . A set of recombinant plasmids has been obtained which can determine the synthesis of A and B individual subunits and are suitable for constructing immunotoxins by gene engineering . The diphtheria toxin of 62 kDa synthesized in E . coli has enzymatic activity and reacts with antitoxin sera . Some sites for E . coli proteases are present in tox-specific polypeptides. Chem Phys Lipids, 1990 Jun, 54(3-4), 215 - 20 Mass spectrometric analysis of O-peracetylated derivatives of 1-monomycoloylglycerol isolated from Corynebacterium pseudotuberculosis, Rhodococcus rhodochrous and R . lentifragmentus; Ioneda T; 1-Monomycoloylglycerols from Corynebacterium pseudotuberculosis, Rhodococcus rhodochrous and R . lentifragmentus were reacted with acetic acid ahydride in the presence of pyridine . On infrared spectra the reaction products showed a sharp characteristic absorption of the acetyl ester group at 1235 cm-1; the hydroxyl group absorption (3400 cm-1) was absent . O-Peracetylated monomycoloylglycerols were analyzed by mass spectrometry under electron impact mode . The most common and representative peaks were associated to the following remarkable fragments: (a) peak at m/z 159 represented the backbone of the glycerol unit of peracetylated monomycoloylglycerols; it constituted the diagnostic and base peak of that group of compounds; (b) peak representing the glycerol moiety together with the alpha-subunit of the mycolic acid moiety; it gave the size of the chain length of the hydrocarbon side chain of that mycolic acid; and (c) a series of peaks of acylium ions minus 60 mass units (acetic acid) indicated the size of the chain length of the esterified mycolic acid . Therefore, acetylation of monomycoloylglycerol becomes the derivatized compound suitable for mass spectrometry . Moreover, the derivatives are more thermostable and resistant to pyrolysis . By associating these properties, separation and identification of homologs of monomycoloylglycerols are further expected. Immunobiology, 1990 Jun, 180(4-5), 405 - 18 Modulation of Forssman glycosphingolipid expression by murine macrophages: coinduction with class II MHC antigen by the lymphokines IL4 and IL6; von Kleist R et al.; In contrast to murine spleen M phi, resident peritoneal M phi from health mice express very little Forssman glycolipid antigen (Fo) . The following experiments suggest that Fo expression by peritoneal M phi may be associated with inflammation . Balb/c and CBA/J mice were given inflammatory stimuli by i.p . injection of live BCG, thioglycollate (TG), Corynebacterium parvum (CP), proteose peptone (PP), or LPS . Control animals received pyrogen-free saline . Expression of Fo and Ia antigen by peritoneal M phi was determined by immunofluorescence after 4 d . Application of TG or CP led to an up to 30-fold increase in Fo+, Ia+ double positive M phi over that in control animals . LPS caused mainly an increase in the percentage of double-positive M phi, whereas no effects were seen in BCG or PP treated animals . To clarify the possible involvement of cytokines in this process and to identify these, the effects of LPS and various cytokines on in vitro induction of Fo and Ia expression were studied in further experiments . LPS, IL6, and IL4 caused induction of up to 15% Fo+ and Ia+ M phi after a 4 d culture period . M phi colony stimulating factor (M-CSF) from lung-conditioned medium was also moderately active . IL1, TNF, and IL2 had no influence, whereas IFN-gamma only induced Ia . For a successful in vitro induction of Fo and Ia, a prior priming of the mice with PP appeared mandatory . This suggests that only M phi of a certain developmental stage can acquire Fo under the influence of the appropriate cytokines . The data may provide the first evidence for cytokine-mediated modulation of a glycolipid antigen of known chemical structure. J Bacteriol, 1990 Jun, 172(6), 3409 - 16 Membrane alteration is necessary but not sufficient for effective glutamate secretion in Corynebacterium glutamicum; Hoischen C et al.; We showed recently that secretion of glutamate in biotin-limited cells of Corynebacterium glutamicum is mediated by carrier systems in the plasma membrane (C . Hoischen and R . Kramer, Arch . Microbiol . 151:342-347, 1989) . In view of the generally accepted hypothesis that glutamate efflux is directly caused by alterations of the membrane, it was necessary to examine the kind of correlation between changes in lipid content and composition of the bacterial membrane and glutamate secretion activity . Two new experimental approaches were used . (i) Changes in lipid content and composition were analyzed in glutamate-producing cells which were forced to switch to nonproducers by addition of biotin in a short-term fermentation . (ii) The time courses of both the fatty acid or phospholipid composition and the efflux activity were analyzed within the first minutes of the switch from high to low secretion activity . The following results were obtained . (i) The time course of the change in fatty acid or phospholipid content and composition was not related to the change in secretion behavior . (ii) There was no specific fatty acid or phospholipid compound which regulated glutamate efflux . (iii) High efflux activity could only be induced when the total lipid content of the membrane was reduced . (iv) Although consistently correlated to high secretion activity, membrane alteration was never a sufficient prerequisite for glutamate efflux in C . glutamicum. Agric Biol Chem, 1990 Jun, 54(6), 1491 - 8 Leucine dehydrogenase from Corynebacterium pseudodiphtheriticum: purification and characterization; Misono H et al.; Leucine dehydrogenase {EC 1.4.1.9} was purified to homogeneity from Corynebacterium pseudodiphtheriticum ICR 2210 . The enzyme consisted of a single polypeptide with a molecular weight of about 34,000 . Stepwise Edman degradation provided the N-terminal sequence of the first 24 amino acids, and carboxypeptidase Y digestion provided the C-terminal sequence of the last 2 amino acids . Although the enzyme catalyzed the reversible deamination of various branched-chain L-amino acids, L-valine was the best substrate for oxidative deamination at pH 10.9 and the saturated concentration . The enzyme, however, had higher reactivity for L-leucine, and the kcat/Km value for L-leucine was higher than that for L-valine . The enzyme required NAD+ as a natural coenzyme . The NAD+ analogs 3-acetylpyridine-NAD+ and deamino-NAD+ were much better coenzymes than NAD+ . The enzyme activity was significantly reduced by sulfhydryl reagents and pyridoxal 5'-phosphate . D-Enantiomers of the substrate amino acids competitively inhibited the oxidation of L-valine. Biochem Biophys Res Commun, 1990 May 16, 168(3), 1034 - 40 Inducible cytosolic enzyme activity for the production of nitrogen oxides from L-arginine in hepatocytes; Billiar TR et al.; The in vivo conditions needed for the induction of nitrogen oxide synthesis by hepatocytes were determined . Hepatocytes obtained from rats injected with killed Corynebacterium parvum spontaneously produced NO2(-)+NO3- in culture and were found to contain cytosolic enzyme activity for nitrogen oxide synthesis . The enzyme activity required both L-arginine and NADPH, and was not found in hepatocytes obtained from normal rats or rats injected with lipopolysaccharide (LPS) alone . In contrast, nonparenchymal cells were stimulated to synthesize NO2(-)+NO3- by LPS . These results show the presence of inducible cytosolic enzyme activity for nitrogen oxide synthesis in hepatocytes, which is distinct from nonparenchymal cell NO . synthesis. J Am Vet Med Assoc, 1990 May 15, 196(10), 1609 - 13 Differential antibody responses to Corynebacterium pseudotuberculosis in sheep with naturally acquired caseous lymphadenitis; Ellis JA et al.; Enzyme-linked immunosorbent assays (ELISA) with Corynebacterium pseudotuberculosis cell wall and bacteria-free supernatant with exotoxin preparations as antigens, and hemolysis inhibition tests were used to detect antibodies in the sera of adult range sheep with naturally acquired caseous lymphadenitis (CL) . The extent and severity of lesions were quantitated on the basis of a lesion score, derived from an examination of the carcass (peripheral lymphoid tissue) and viscera (including internal lymphoid tissue) at the time of slaughter . The overall prevalence of C pseudotuberculosis-positive CL lesions in 104 sheep was 31.7% . The cell wall ELISA detected antibodies in 96.9% (32/33) of sheep with C pseudotuberculosis-positive CL lesions . The exotoxin ELISA detected antibodies in 84.8% (28/33) of positive sheep in the same group . Both ELISA resulted in a high number of apparent false-positives, with 64.7% and 49.2%, respectively, positive optical density (OD) values in sheep with no gross CL lesions and no apparent C pseudotuberculosis infection . There was no significant relationship between the extent of lesion development (lesion score) and OD values in both cell wall (r = 0.472) and exotoxin (r = 0.464) ELISA . Similarly, there was no significant relationship between the titer of antitoxin antibodies, as measured by the hemolysis inhibition test, and the extent of disease . These investigations indicate that those ELISA that use crude C pseudotuberculosis antigens are of questionable utility in the field, where C pseudotuberculosis infection is endemic in many sheep populations . Furthermore, these studies suggest that antibodies that are reactive with components of C pseudotuberculosis and that develop in response to infection may have little impact on the recovery of the host. Med J Aust, 1990 May 7, 152(9), 458, 461 - 3 Endocarditis associated with prosthetic cardiac valves; Chen SC et al.; Clinical features, microbiology, therapy and outcome of 26 episodes of prosthetic valve endocarditis occurring at Westmead Hospital from 1979 to 1989 were examined retrospectively . Presentation with a new or changed cardiac murmur was associated with early onset infection (within 12 months of prosthetic valve insertion; P = 0.0033) . Corynebacteria were the commonest cause of early onset endocarditis (4 of 11 episodes) and Streptococcus viridans of late onset endocarditis (4 of 15 episodes) . Nine of 11 episodes responded to antimicrobial therapy and 12 of 15 to medical-surgical therapy . There was a trend towards increased mortality in patients with early onset endocarditis presenting with a new or changed cardiac murmur (4 of 9 v . 1 of 17, P = 0.068), suggesting early surgery should be considered in this group . Analysis of antibiograms and published reports indicated that vancomycin and an aminoglycoside should be recommended as empirical therapy for endocarditis occurring 12 to 18 months after prosthetic valve insertion. Pathol Biol (Paris), 1990 May, 38(5), 355 - 7 {Activity of cefuroxime against bacterial strains isolated from acute otitis media}; Simonet M et al.; The acute otitis media is a frequent infantile disease and, in 80% of cases, a bacterial strain can be isolated from the otorrhoea . Haemophilus influenzae and Streptococcus pneumoniae are the two major species isolated from auricular exudate, and represent two thirds of all isolated strains, with the others comprising Staphylococcus aureus, Branhamella catarrhalis, Pseudomonas aeruginosa, Enterobacteriaceae and corynebacteria . The treatment of this disease is based principally on beta-lactams (aminopenicillins, cephalosporins) administered by the oral route . Cefuroxime is a cephalosporin which is absorbed via the digestive tract in the form of cefuroxim-axetil . The activity of this compound was studied against 210 strains isolated from otorrhoea, collected from children who presented an acute otitis media during the first half of 1989 . These strains were: 112 strains of H . influenzae, of which 23 produced a beta-lactamase; 21 strains of Streptococcus pneumoniae; 3 strains of Streptococcus pyogenes; 10 strains of Branhamella catarrhalis of which 9 produced a beta-lactamase; 18 strains of S . aureus; 14 strains of Enterobacteriaceae, and 32 strains of corynebacteria . The minimal inhibitory concentration (MIC) of cefuroxime-axetil was measured by dilution in agar . The MICs of cefuroxime against H . influenzae were low and similar (MIC 50 = 1 mg/l; MIC 90 = 1 mg/l) regardless of whether the strain secreted a beta-lactamase . Overall, 90% and 98% of the 210 strains tested here were inhibited by 1 and 4 mg/l of cefuroxime respectively . These results show that the antibacterial spectrum of cefuroxime-axetil appears to be ideally suited to the bacterial strains isolated from acute otitis media. Rev Infect Dis, 1990 May-Jun, 12(3), 412 - 5 Unusual pathogens in narcotic-associated endocarditis; Szabo S et al.; We report the cases of three adults with a history of intravenous drug abuse who developed endocarditis caused by Corynebacterium xerosis, Neisseria subflava, and Neisseria flavescens, respectively . No cases of endocarditis caused by C . xerosis or N . flavescens and only one case caused by N . subflava have previously been reported in association with narcotic addiction . The prominent clinical features in all patients included poor response to antibiotic therapy, persistent fever, and major embolic events . Stigmata of infection with human immunodeficiency virus, as manifested by oral candidiasis, cervical lymphadenopathy, and serologic evidence, were present in two of the three patients . At our institution, where Staphylococcus aureus remains the most frequent etiologic agent of narcotic-associated endocarditis, the occurrence of these three cases in a 9-month period is striking . We speculate that infection with human immunodeficiency virus may play a role in the pathogenesis of endocarditis caused by these unusual organisms. Mol Biol Evol, 1990 May, 7(3), 247 - 54 Contrasting evolutionary histories of chloroplast thioredoxins f and m; Hartman H et al.; Fourteen thioredoxin sequences were used to construct a minimal phylogenetic tree by using parsimony . The bacterial thioredoxins clustered into three groups: one containing the photosynthetic purple bacteria, Escherichia and Corynebacterium; a second containing the photosynthetic green bacterium, Chlorobium; and a third containing cyanobacteria . These groupings are similar to those generated from earlier 16s RNA analyses . Animal thioredoxins formed a fourth group . The two thioredoxins of chloroplasts (f and m) showed contrasting phylogenetic patterns . As predicted from prior studies, spinach chloroplast thioredoxin m grouped with its counterparts from cyanobacteria and eukaryotic algae, but, unexpectedly, thioredoxin f grouped with the animal thioredoxins . The results indicate that, during evolution, thioredoxin m of contemporary photosynthetic eukaryotic cells was derived from a prokaryotic symbiont, whereas thioredoxin f descended from an ancestral eukaryote common to plants and animals . The findings illustrate the potential of thioredoxin as a phylogenetic marker and suggest a relationship between the animal and f-type thioredoxins. J Clin Microbiol, 1990 May, 28(5), 913 - 21 Protein G-based enzyme-linked immunosorbent assay for anti-MPB70 antibodies in bovine tuberculosis; Harboe M et al.; MPB70 is a highly species specific protein which is secreted from Mycobacterium bovis during culture . To investigate whether antibodies against MPB70 can be used as an indicator of infection with M . bovis, an enzyme-linked immunosorbent assay was developed, based on the use of biotinylated protein G, to provide a common indicator for antibody formation in different species . During experimental infection with M . bovis in cattle, a characteristic pattern of anti-MPB70 antibody production was observed with an initial flat plateau followed by a marked rise 18 to 20 weeks after infection . Skin testing with bovine tuberculin purified protein derivative (PPD), which was shown to contain antibody-reactive MPB70, was a potent stimulator of antibody production in infected animals . In experimentally infected cattle, we observed an inverse relationship between antibody activity and delayed-type hypersensitivity skin test reactions . In natural M . bovis infections, skin testing with PPD was also a potent stimulator of anti-MPB70 formation . Comparison between the enzyme-linked immunosorbent assay for antibodies to MPB70 and that for antibodies to the widely cross-reacting M . bovis BCG antigen 85B in animals with M . bovis, Mycobacterium avium, Mycobacterium paratuberculosis, and Corynebacterium pseudotuberculosis infections showed that formation of antibody to MPB70 was highly specific for infection with M . bovis . The use of an MPB70-containing PPD preparation for skin testing followed by this anti-MPB70 assay is a highly specific indicator of M . bovis infection . Adjustment of the test conditions is expected to provide an increased sensitivity of the procedure for the diagnosis of natural M . bovis infections. Pathol Biol (Paris), 1990 May, 38(5), 420 - 5 {Sensitivity curves of multiply resistant corynebacteria JK and D2}; Rauturier M et al.; Corynebacteria groups JK and D2 are opportunistic pathogens . They are sometimes responsible for severe infections, especially in immunocompromised patients . They are resistant to many antibiotics . Under these conditions it seemed important to discover which antibiotics among those regularly active, have a killing effect, what their bactericidal speed is, and how they react to combination with other antibiotics . Glycopeptids have a killing effect within 24 hours . Pristinamycin, rifampin, fusidic acid, when active, are bacteriostatic . Their combination with vancomycin is indifferent but in one case the combination vancomycin + rifampin is synergistic. J Dent Res, 1990 May, 69(5), 1131 - 7 pH regulation of urease levels in Streptococcus salivarius; Sissons CH et al.; Potential mechanisms for regulation of urease levels in Streptococcus salivarius were examined, including: induction by urea, nitrogen or carbon source repression, and effects of pH and CO2 (because CO2 enrichment enhanced urease detection on urea agar plates) . Regulation by either pH or CO2 was confirmed by comparison of the urease accumulation pattern during anaerobic growth under CO2 with that under N2 . Under CO2, there was an initial buffering plateau at pH 6.2 and a rate of Streptococcus salivarius urease accumulation three-fold that under N2, with a pH 7.6 plateau . With both gas phases there was also an increase in the rate of urease appearance coincident with the decrease in medium pH following the pH plateau . The effects of pH, CO2, and HCO3- on urease levels and on growth were separately assessed by culture in media containing 0, 25, 100 mmol/L KHCO3 buffered at different pH levels . There was an inverse relationship between the logarithm of the urease level after 24-hour growth and the pH during growth-the urease specific activity was 100-fold higher at pH 5.5, compared with pH 7.0 and above . HCO3-/CO2 (100 mmol/L) had little effect on urease levels, but was essential for growth at pH 5.5 . There was no significant urease induction by urea, or repression by ammonia or glucose . There was also evidence of pH regulation of urease levels in some staphylococci, Klebsiella pneumonia, and Corynebacterium renale, but not in Actinomyces naeslundii and several other species . We conclude that the external pH is a major factor regulating urease levels in S . salivarius and possibly some other species-a mechanism equivalent to urease repression by OH-. Cas Lek Cesk, 1990 Apr 20, 129(16), 498 - 500 {Monitoring natural killer cell activity in the blood of patients with malignant pleural effusion after intrapleural administration of Corynebacterium parvum}; Marel M et al.; In a group of 14 patients with a malignant pleural effusion treated by intrapleural administration of Corynebacterium parvum, in addition to other treatment of the generalized malignant process the activity of Nk cells was investigated before administration, on the 7th and 21st day after instillation of the vaccine . The mean rise of this activity was significant at the 5% level of significance . The activity of Nk cells was equally significantly influenced by age . The survival of patients, although longer in patients who responded positively, was not significantly correlated with the increased activity of Nk cells . Its increase was observed more frequently in the group of patients given the English preparation Coparvax . In the discussion some aspects of the relationship of Nk activity and anti-tumourous immunity are discussed. Br J Ophthalmol, 1990 Apr, 74(4), 247 - 8 Mixed haematogenous endophthalmitis caused by Candida albicans and CDC fermentative corynebacterium group A-4; Barker C et al.; We report a case of mixed haematogenous endophthalmitis in which Candida albicans and Centres for Disease Control (CDC) corynebacterium group A-4 were isolated together from an aspirate of vitreous humour. Am J Infect Control, 1990 Apr, 18(2), 82 - 5 Efficiency of skin sterilization for a venipuncture with the use of commercially available alcohol or iodine pads; Choudhuri M et al.; Skin sterilization for a venipuncture is routinely done with commercially available alcohol or iodine pads . Selection of the antiseptics, alcohol or iodine, however, in most situations has been made on the basis of very little scientific data . With many patients with granulocytopenia who are undergoing venipunctures, the choice of antiseptic may be an important factor in preventing infections . We investigated two widely and commercially available disinfectant pads, alcohol and iodine, in the efficacy of skin sterilization . Seventy subjects (35 adults and 35 children) were randomly selected for this study . A designated area of the right and left forearm was sterilized either with alcohol or with an iodine pad in a predetermined uniform fashion . Specimens were obtained for cultures before and after sterilization . The bacterial cultures were performed with the use of blood agar plates and trypticase soy broth . For data analysis growth of any organisms on agar plates or trypticase soy broth after sterilization was interpreted as a sterilization failure . The iodine swab was significantly more efficient than the alcohol swab; the former yielded an 80% sterilization rate whereas the latter resulted in a rate of 61% (p less than 0.02) . If, however, the growth only in agar plates was compared, the alcohol wipe yielded no growth in 83% and iodine in 84%, virtually identical success rates . Bacillus spp . predominated the residual organisms after either the alcohol or the iodine wipe . A variety of other organisms, however, including Staphylococcus and Corynebacterium spp., grew after alcohol but not after iodine sterilization.(ABSTRACT TRUNCATED AT 250 WORDS) J Immunol, 1990 Apr 1, 144(7), 2725 - 9 Microbiostatic effect of murine-activated macrophages for Toxoplasma gondii . Role for synthesis of inorganic nitrogen oxides from L-arginine; Adams LB et al.; Recent studies show the importance of a single amino acid, L-arginine, as a necessary substrate for activated macrophage-mediated cytotoxic activity for tumor target cells and microbiostatic function for Cryptococcus neoformans . The present studies were carried out to determine the role of the L-arginine-dependent macrophage effector function on the microbiostatic effects of activated macrophages on the obligate intracellular protozoan, Toxoplasma gondii . A guanidino methylated derivative of L-arginine, NGmonomethyl-L-arginine (NGMMA), a competitive inhibitor of the L-arginine-dependent effector pathway, virtually abolished the normally potent microbiostatic effect of macrophages for Toxoplasma gondii after activation of the macrophages in vitro by IFN-gamma and LPS or in vivo by i.p . injection of killed Corynebacterium parvum . Addition of supplemental L-arginine to the culture medium overcame the capacity of NGMMA to block activated macrophage-mediated microbiostasis of Toxoplasma . The ability of NGMMA to inhibit the microbiostatic capacity of activated macrophages for Toxoplasma gondii correlated with almost total inhibition of synthesis of nitrite, nitrate, and L-citrulline from L-arginine . Therefore, as is the case for tumor target cells and C . neoformans, the synthesis of inorganic nitrogen oxides from a terminal guanidino nitrogen atom of L-arginine appears to be essential for murine cytotoxic activated macrophage mediated microbiostatic capacity for T . gondii. Pathol Biol (Paris), 1990 Apr, 38(4), 302 - 6 {Antibiotherapy in the patient with granulocytopenic cancer}; Van Der Auwera P et al.; Granulocytopenia is the leading alteration of the natural host defense mechanisms, whether caused by an underlying disease or associated with anticancer chemotherapy and radiation therapy . Severe granulocytopenia predisposes to septicemia which is now more often due to Gram positive than to Gram negative bacteria . The empiric therapy of febrile episodes with rapidly bactericidal antibiotics has dramatically modified the prognosis of septicemia . The optimal treatment remains controversial although the usual regimen include both a cephalosporin (or a large spectrum penicillin) with an aminoglycoside . The empiric treatment with a specific anti-Gram positive antibiotic such as vancomycin does not modify prognosis, adds significant side-effects and increases the cost . Monotherapy has been associated with the need for treatment modification in 30-80% of the episodes depending on the type of infection (fever of unknown origin, clinically or microbiologically documented infection) . The patients who remain febrile despite adequate antibacterial empiric treatment beneficiate of an empiric antifungal treatment . Care should be taken about the recent emergence of vancomycin-resistant Staph . haemolyticus, Corynebacterium JK and non-JK, and non-aeruginosa pseudomonas (only susceptible to cotrimoxazole) . More fungal infections are observed with a significant emergence of non-albicans Candida, dermatophytes and filamentous fungi (P . boydii, Fusarium, ...) associated with disseminated infections. J Bacteriol, 1990 Mar, 172(3), 1256 - 61 Cloning, nucleotide sequence, and expression in Escherichia coli of the phospholipase D gene from Corynebacterium pseudotuberculosis; Hodgson AL et al.; The phospholipase D (PLD) gene from Corynebacterium pseudotuberculosis has been cloned, sequenced, and expressed in Escherichia coli . Analysis of DNA sequence data reveals a major open reading frame encoding a 31.4-kilodalton protein, a size consistent with that estimated for the PLD protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Comparison of these data with the amino-terminal protein sequence indicates that the mature PLD protein is preceded by a 24-residue signal sequence . Expression of the PLD gene in E . coli is initiated from the corynebacterial promoter, and the resulting protein has sphingomyelinase activity . Primer extension mapping localized the 5' end of the PLD gene mRNA to a site 5 to 7 base pairs downstream of a region similar to the consensus sequence for E . coli promoters . Northern and Southern blot analyses suggest that the gene is transcribed from mRNA approximately 1.1 kilobases in length and that it is present in a single copy within the C . pseudotuberculosis genome. J Am Vet Med Assoc, 1990 Mar 1, 196(5), 760 - 2 Hypertrophic osteopathy in a dog with a chronic lung abscess; Hesselink JW et al.; A 2-year-old dog was examined because of gradual lameness of all 4 limbs and weight loss . Hypertrophic osteopathy was diagnosed . Radiography revealed a mass in the caudal lobe of the right lung . At necropsy, the mass was determined to be a chronic abscess . Corynebacterium pyogenes was cultured from the lesion . Although hypertrophic osteopathy in dogs is commonly associated with a thoracic lesion, most often neoplastic, the chronic lung abscess in this dog might have been formed as a result of a paralaryngeal abscess that was treated 3 months before the onset of the lameness. J Gen Microbiol, 1990 Mar, 136 ( Pt 3), 515 - 9 Characterization and partial purification of L-asparaginase from Corynebacterium glutamicum; Mesas JM et al.; A high L-asparaginase (L-asparagine amidohydrolase: EC 3.5.1.1) activity was found under conditions of lysine overproduction in cultures of Corynebacterium glutamicum . L-Asparaginase was purified 98-fold by protamine sulphate precipitation . DEAE-Sephacel anion exchange, ammonium sulphate precipitation and Sephacryl S-200 gel filtration . The asparaginase protein was subjected to PAGE under non-denaturing conditions, identified by an in situ reaction and eluted from the gel in an active form . The estimated Mr from gel filtration and SDS-PAGE was 80,000 . The L-asparaginase activity was inhibited by the L-asparagine analogue 5-diazo-4-oxo-L-norvaline . Neither D-asparagine nor L-glutamine was a substrate for the enzyme . L-Asparaginase was produced constitutively: its role may be that of an overflow enzyme, converting excess asparagine into aspartic acid, the direct precursor of lysine and threonine. Taehan Chikkwa Uisa Hyophoe Chi, 1990 Mar, 28(3), 299 - 301 {A microbiologic study on anaerobic bacteremia after dental extraction}; Lee JG et al.; It has been known for many years that tooth extraction can cause bacterial dissemination into blood stream, that is, bacteremia . Besides endocarditis--causing bacteria, alpha-hemolytic streptococci, anaerobes are involved in the post- extraction bacteremia . They are of clinical importance because they can cause abscesses in the various parenchymatous organs such as the liver, lung, and brain through anachoresis . The purpose of this study is to establish the types of species of anaerobes associated with post--extraction bacteremia, and to give the baseline to the succeeding study of sensitivity tests of various antibiotics, thus clinicians can chose the right ones . 10ml of blood was sampled pre- and post- operatively from 32 patients and incubated in anaerobic chamber . The followings are the results of this study . 1 . All the preoperative blood samples showed negative culture . 2 . Postoperative blood samples from 8 out of 10 patients (80%) in group 1, from 1 out of 15 (73.3%) in group 2, and 2 out of 7 (28.6%) in group 3 yielded anaerobic and facultative species . 3 . Organisms detected were species belonging to the genera Streptococcus, Bacteroides, Actinomyces, Staphylococcus, Peptostreptococcus, Peptococcus, Propionebacterium, Fusobacterium, nonenterococcus, and Corynebacterium. Zhonghua Yu Fang Yi Xue Za Zhi, 1990 Mar, 24(2), 77 - 9 {Survey of microbes in hospital environment}; Duan FR; Bacterial count and species distribution in hospital ward was investigated . It was shown that the in air total bacterial count was maximum in the second season of the year and the number of Streptococcus above the standard was in the first season . Bacteria in the air of the ward by number was in this order: Micrococcus tetragenus, Staphylococcus, Bacillus subtilis, Streptococcus and Corynebacterium . According to the number of bacteria on the equipment and door handle of the ward, the order, was B . subtilis, Pseudomonas and Staphylococcus . Disinfectant solution used for tools and instruments was proved to be effective . The observation is useful to hospital administration. J Bacteriol, 1990 Mar, 172(3), 1663 - 6 High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria; Schafer A et al.; We report on the mobilization of shuttle plasmids from gram-negative Escherichia coli to gram-positive corynebacteria mediated by P-type transfer functions . Introduction of plasmids into corynebacteria was markedly enhanced after heat treatment of the recipient cells . High-frequency plasmid transfer was also observed when the restriction system of the recipient was mutated . On the basis of our data, we conclude that efficient DNA transfer from gram-negative to gram-positive bacteria, at least to coryneform bacteria, is conceivable in certain natural ecosystems. J Biotechnol, 1990 Mar, 13(4), 257 - 66 Microbial glycolipid production under nitrogen limitation and resting cell conditions; Kim JS et al.; Rhodococcus erythropolis is able to synthesize an anionic trehalose-2,2',3,4-tetraester during cultivation on n-alkanes . Preconditions for an overproduction are nitrogen limitation, temperature- and pH-shift . The optimum carbon source was technical grade n-C-10, which led to 0.35 g g-1 of glycolipid per n-alkane . Electron microscopical observations showed that n-C-14,15 (technical grade) grown cells contained numerous lipid inclusions in contrast to n-C-10 (technical grade) grown cells . Nocardia corynebacteroides synthesizes a novel pentasaccharide lipid and as size products small amounts of trehalose-corynomycolates . Optimum precursors for overproduction are n-alkanes from n-tetradecane to n-hexadecane with yields in the range of 0.17 g g-1 of glycolipid per carbon source. Microb Pathog, 1990 Feb, 8(2), 157 - 62 Changes in ovine erythrocyte morphology due to sphingomyelin degradation by Corynebacterium pseudotuberculosis phospholipase D; Brogden KA et al.; Corynebacterium pseudotuberculosis produces a sphingomyelin-specific phospholipase D exotoxin that is a major determinant in the pathogenesis of caseous lymphadenitis . We assessed the changes induced in the morphology and sphingomyelin concentration of ovine erythrocytes after incubation with broth culture filtrates or purified exotoxin . Sphingomyelin was hydrolysed nearly in parallel with shape change . Morphologically, spherostomatocytes were seen initially, and later these contained numerous vacuoles at the periphery . Vacuoles seen in thin sections with transmission electron microscopy appeared as pits with scanning electron microscopy . Pitting became progressively worse with time, leading to extensive scalloping of the membrane surface . Chemically, significant decreases (P less than or equal to 0.05) in erythrocyte membrane sphingomyelin content and significant increases (P less than or equal to 0.05) in the content of glycerophospholipids (i.e . phosphatidylglycerol) were observed in erythrocytes incubated with broth culture filtrates or purified exotoxin . These alterations may contribute to the pathophysiology of acute infections induced by C . pseudotuberculosis. Immunobiology, 1990 Feb, 180(2-3), 124 - 37 Effects of stimulated or immunologically activated macrophages on the induction of immune responses to Listeria monocytogenes; Yoshikai Y et al.; The influences of peritoneal macrophages induced by proteose peptone, Corynebacterium parvum (C . parvum) or Bacillus Calmette Guerin (BCG) on the initiation and development of immune responses and protection against Listeria monocytogenes infection were studied in mice . Mice treated intraperitoneally (i.p.) with proteose peptone 4 days previously showed much the same level of protection against an intraperitoneal infection with Listeria as untreated mice . Mice treated i.p . with C . parvum 4 days previously, of which peritoneal macrophages had increased abilities for intracellular killing of Listeria and O2- generation as compared with peptone-elicited macrophages, exhibited an enhanced resistance against the listerial infection . The degree of immune responses, as assessed by delayed footpad reaction (DFR), was rather depressed in these mice because C . parvum-activated macrophages acting as scavenger cells reduced the amount of effective antigenic stimulation . BCG-activated peritoneal macrophages from mice treated i.p . with BCG 14 days previously showed a strong ability for antigen presentation in correlation with increases in the number of Ia-bearing macrophages and in the level of interleukin 1 (IL 1) production . These mice showed an early appearance of DFR response and a markedly enhanced resistance against the listerial infection . These results suggested that the differences in macrophage activities as scavenger cells, cytokine-secreting cells and antigen presenting cells may account for the differences in the responsiveness against listerial infection in peptone-, C . parvum- and BCG-treated mice. J Dairy Sci, 1990 Feb, 73(2), 351 - 6 Growth of Corynebacterium bovis in mammary secretions during physiological transitions of the bovine mammary gland; Oliver SP et al.; An in vitro microassay was used to evaluate growth of five strains of Corynebacterium bovis in mammary secretions collected from quarters of five Holstein cows at 0, 14, and 28 d of involution, at parturition, and 14 d after parturition . Variation in growth among different strains of Corynebacterium bovis was observed . Corynebacterium bovis grew well in mammary secretions obtained at the last milking of lactation, at parturition, and 14 d after parturition . However, growth of four strains of Corynebacterium bovis in mammary secretions obtained at 14 and 28 d of involution was reduced significantly . In contrast, a streptomycin-resistant strain of Corynebacterium bovis grew well in mammary secretions obtained during involution . These data suggest that mammary secretions support growth of Corynebacterium bovis during lactation but inhibit growth during the nonlactating period . Inhibition of growth in secretions collected during the nonlactating period may be associated with the high rate of spontaneous elimination of Corynebacterium bovis intramammary infection from cessation of milking to parturition . Conversely, enhanced growth in milk may be related to persistent Corynebacterium bovis intramammary infections during lactation. Histopathology, 1990 Feb, 16(2), 187 - 9 Diphtheria with a difference--a rare Corynebacterium fatality with associated apoptotic cell death; Leek MD et al.; A case of infection with Corynebacterium ulcerans (C . ulcerans), resulted in the sudden death of a previously healthy 73-year-old woman . Death from Corynebacterium diphtheriae (C . diphtheriae) infection is well-documented . Fatalities following infection with C . ulcerans are unreported; this would appear to be the first documented death due to such infection. Eur Respir J, 1990 Feb, 3(2), 240 - 2 Human pulmonary infection with Corynebacterium equi; Egawa T et al.; A 28 year old man with no underlying disease developed a cavity and multiple nodules in the lung from which Corynebacterium equi was isolated . He experimented with organic solvents and microorganisms including Corynebacterium species for several years . Computed tomography of his pulmonary lesions revealed that these nodules were related to the bronchial tree . Histologically, the lesions were compatible with nonspecific inflammatory changes . The clinician must suspect the pulmonary infections with Corynebacterium species even if the patient has no underlying disease. Am J Vet Res, 1990 Feb, 51(2), 200 - 2 Effect of muramyl dipeptide on immunogenicity of Corynebacterium pseudotuberculosis whole-cell vaccines in mice and lambs; Brogden KA et al.; Colostrum-deprived lambs and CF1 mice were vaccinated with water-in-oil emulsion vaccines containing nonviable whole cells (WC) of Corynebacterium pseudotuberculosis with and without muramyl dipeptide (MDP) . Efficacy of vaccines was determined from the survival of mice and lesions in lambs after IV injection of 10(4) colony-forming units of C pseudotuberculosis . In mice, protection was related to the concentration of WC in the vaccine . At 50, 100, or 150 micrograms of WC, protection was good (78.8%) . At 10 or 25 micrograms of WC, protection was considerably less (54.7%) . At high WC concentrations, protection could only be moderately increased to 82.3% with high (50 and 100 micrograms) concentrations of MDP or increased to 90% protection with low (5 and 10 micrograms) concentrations of MDP . At low WC concentrations, protection significantly decreased to 32% (P less than 0.025) with high concentrations of MDP, but significantly increased to 72.5% (P less than 0.025) with low concentrations of MDP . Therefore, the amount of protection with lower concentrations of WC and MDP was comparable with the amount of protection with higher concentrations of WC without MDP . In lambs, high prechallenge antibody titers (geometric mean titers from 5.1 to 5.4 by day 35) were observed after vaccination with WC . Protection and vaccination site abscesses in lambs were related to the concentration of WC and MDP . Pulmonary or vaccination site abscesses were not observed in 4 of 4 lambs vaccinated with 1 mg of WC + 50 micrograms of MDP. J Antimicrob Chemother, 1990 Feb, 25 Suppl B, 1 - 5 The antimicrobial activity of fusidic acid; Verbist L; Fusidic acid, a fusidane that interferes with protein synthesis via the translocase enzyme, is mainly notable for its activity against staphylococci, coagulase-positive and negative, whether or not they are resistant to methicillin and related penicillins . It is also active against corynebacteria and against many genera of strict anaerobes and microaerophiles . Mutants showing resistance, by more than one mechanism, may readily be selected in vitro . Combination of fusidic acid with other antibiotics generally gives addition or indifference, but also delays emergence of resistant mutants. Chem Phys Lipids, 1990 Feb, 52(3-4), 259 - 62 Effect of low mole fraction of trehalose dicorynomycolate from Corynebacterium diphtheriae on water permeability and electrical capacitance of lipid bilayer membranes; Imasato H et al.; The effect of incorporation of different proportions of trehalose dicorynomycolates (TDC) into lecithin bilayer membranes was studied . It was found that TDC, induces a 14% decrease of water osmotic permeability (42.6 +/- 3.9 to 36.8 +/- 2.7 microns/s) at 1.6 mole%, suggesting that this substance leads to an increase of the degree of packing of the constituent lipid molecules . A condensing effect of TDC was also apparent from membrane electrical capacitance (Cm) measurement . By incorporating TDC into bilayer membranes, the value of Cm experienced a decrease of 29% at 1.6% mole fraction . This finding was taken to reflect an increase in membrane thickness, known in many examples, to be related to the condensing effect. J Med Microbiol, 1990 Feb, 31(2), 137 - 49 A pyrolysis-mass spectrometry study of Corynebacterium spp; Hindmarch JM et al.; Clinical (66) and collection (38) strains of Corynebacterium spp., including C . jeikeium and CDC group D2, and of Listeria monocytogenes were examined . Conventional characters used in species identification were assessed by a microbiochemical method, and pyrolysis-mass spectrometry (Py-MS) was performed with a Horizon Instruments PYMS 200X . Classification based on Py-MS data yielded clusters that corresponded with species identification and classification groups from conventional data . One small group of clinical strains, homogeneous in conventional tests and Py-MS, comprised isolates from sputum samples from patients undergoing ventilation; they were similar to collection strains of C . renale and C . striatum; the latter species has been implicated in chest infection . Another group, similar to C . minutissimum in both systems, comprised clinical strains isolated from urogenital specimens . L . monocytogenes strains were clearly distinct from Corynebacterium spp . Groups comprising CDC D2 strains and C . jeikeium were resolved, and were similar to other Corynebacterium spp . Two collection strains of C . xerosis were distinct in conventional tests and Py-MS. Enferm Infecc Microbiol Clin, 1990 Feb, 8(2), 88 - 90 {Corynebacterium jeikeium septicemia}; Sanz JC et al.; An adult male with non-Hodgkin lymphoma, who had been admitted to the hospital for more than one month and had received previous antibiotic therapy, developed Corynebacterium jeikeium septicemia . The organism was isolated in four blood cultures and it was multirresistant, being only sensitive to vancomycin, ciprofloxacin, norfloxacin and rifampin . The patient improved clinically and was bacteriologically cured with intravenous vancomycin therapy. Nippon Juigaku Zasshi, 1990 Feb, 52(1), 11 - 8 Cloning and expression of a pili gene of Corynebacterium renale in Escherichia coli; Abe S et al.; A plasmid gene library of Corynebacterium renale piliated strain No . 109P+ was prepared in Escherichia coli in order to study the chemical structure of the pili of C . renale . Of 3,000 recombinant clones tested, 5 reacted with anti-pili anti-serum . The gene products of these clones reacted with anti-pili monoclonal antibodies 8/4, 5/2 and B20/3 but lacked the reactivity with 13/4 . SDS-PAGE analysis revealed that the expressed protein had a molecular mass of 48 kilodalton and deletion analysis showed that the encoding region for this protein was localized within a 1.4 kilobase gene including a promoter sequence . Immunoelectron microscopy showed that mouse antibodies raised to the expressed protein bound to the entire surface of the pili of C . renale . These results indicate that the cloned gene encodes a major structural protein of C . renale pili. J Immunol, 1990 Jan 15, 144(2), 699 - 706 Characterization of two proteins from Leishmania donovani and their use for vaccination against visceral leishmaniasis; Jaffe CL et al.; Two proteins from Leishmania donovani, dp72 and gp70-2, have been previously utilized to specifically serodiagnose patients with visceral leishmaniasis . The proteins were shown by ELISA and Western blotting with monoclonal and polyclonal antibodies to be present in both stages of the parasite . Antibodies to gp70-2 recognize in promastigotes multiple discrete bands of similar m.w . which are common to several isolates of L . donovani . The total amount of Ag and number of bands observed per isolate is not constant . Lectin blots with Con A show gp70-2 to be a glycoprotein . Dp72 shows pronounced microheterogeneity between isolates of L . donovani . The Brazilian isolates examined appear to possess a lower m.w . form (64,000 or 68,000) of this molecule . No reactions were observed with dp72 and lectins in Western blots; and neither tunicamycin, N-glycanase, endoglycosidase H nor F affected the migration of {35S}-methionine-labeled protein on SDS-PAGE . A mAb against dp72 also cross-reacted in Western blots with a 60-kDa protein in Leishmania major, Leishmania aethiopica, and Leishmania tropica . No reaction was observed between the purified promastigote surface protease (gp63) and either monoclonal or polyclonal antibodies produced to dp72 or gp70-2 . The ability of the pure proteins to provide protection against a challenge by L . donovani amastigotes was examined . BALB/c mice were immunized with gp70-2 and/or dp72 by using Corynebacterium parvum as an adjuvant . Mice immunized with gp70-2 were not protected; however, mice receiving dp72 showed a 81.1% reduction in the liver parasitemia compared with the adjuvant controls. Infect Immun, 1990 Jan, 58(1), 131 - 6 Cloning and expression of the phospholipase D gene from Corynebacterium pseudotuberculosis in Escherichia coli; Songer JG et al.; A toxic phospholipase D (PLD) is putatively involved in pathogenesis of Corynebacterium pseudotuberculosis infections . We report here the cloning and expression of the PLD gene (pld) in Escherichia coli . A cosmid library of DNA from C . pseudotuberculosis biovar ovis isolate Whetten 1 was constructed and screened for PLD-producing recombinants by plating them on LB agar containing sheep erythrocytes and equi factors . One recombinant, designated pCpO1, yielded a gene product which displayed synergistic hemolytic and sphingomyelinase D activities, both of which are characteristic of PLD . Subcloning into pUC19 yielded a recombinant, pCpO50, which contained a 1.8-kilobase insert . Analysis of supernatant fluids and cell extracts of cultures of E . coli(pCpO50) revealed sphingomyelinase activity and a protein of about 31,000 Mr, neither of which were detected in E . coli(pUC19) . The 31-kilodalton protein also reacted with antibodies in serum from a sheep naturally infected with C . pseudotuberculosis, serum which also contained PLD-neutralizing antibodies . When Southern blots of BamHI digests of DNA from biovar ovis and biovar equi isolates of C . pseudotuberculosis were probed with pCpO50, bands of 4.8 and 1.9 kilobases, respectively, were seen, suggesting that the genome organization of pld is different for isolates from the two biovars. Can J Vet Res, 1990 Jan, 54(1), 195 - 7 In vitro susceptibility of selected veterinary bacterial pathogens to ciprofloxacin, enrofloxacin and norfloxacin; Prescott JF et al.; The minimum inhibitory concentrations (MIC) of ciprofloxacin, enrofloxacin, and norfloxacin were tested for approximately ten clinical isolates of each of Actinobacillus pleuropneumoniae, Actinobacillus suis, Actinomyces pyogenes, Corynebacterium pseudotuberculosis, Erysipelothrix rhusiopathiae, Haemophilus parasuis, Haemophilus somnus, Pasteurella haemolytica, Pasteurella multocida, Rhodococcus equi, Streptococcus equi, Streptococcus suis and Streptococcus zooepidemicus . Ciprofloxacin and enrofloxacin had similar activity and were more active than norfloxacin . All isolates had an MIC of 1.0 microgram/mL or less for ciprofloxacin and enrofloxacin, and these drugs had particularly marked activity against the gram-negative bacteria tested. Oncology, 1990, 47(1), 62 - 4 Malignant pleural effusions: meaning of pleural-fluid pH determination; Foresti V et al.; In 36 patients with malignant pleural effusions, we determined the pH and the glucose concentration of the pleural fluid . Twenty-one of 36 patients (58.3%) had a low pH (less than 7.30) and 15 had a normal pH (greater than or equal to 7.30; 7.13 +/- 0.12 vs . 7.37 +/- 0.05; p less than 0.0005) . The patients with low pH had significantly lower glucose concentrations than those with normal pH (2.7 +/- 1.4 vs . 6.3 +/- 2.9 mmol/l; p less than 0.0005) . Twenty-one of 34 patients (61.7%) had a glucose concentration lower than a cut-off value of 4.4 mmol/l; of these, 17 (81%) had a low pH . The mean survival in the low-pH group was 4.8 +/- 4.4 months, whereas the mean survival in the normal-pH group was 5 +/- 8 months (p greater than 0.4) . Twelve of 36 patients (33.3%) were treated with intrapleural Corynebacterium parvum (CBP) injections . Fourteen of 21 low-pH patients (66.6%) survived more than 2 months, and 4 of them are still alive . Six of 15 normal-pH patients (40%) survived more than 2 months, and 1 of them is still alive . Three of the 5 living patients were treated with CBP (2 in the low-pH group and 1 in the normal-pH groups) . Our results confirm that pH and glucose concentrations in the pleural fluid of patients with malignant effusions are frequently low . However, the survival and the response to CBP pleurodesis in patients with low-pH malignant effusions are the same as those in patients with normal-pH malignant effusions. J Clin Microbiol, 1990 Jan, 28(1), 8 - 10 Native valve endocarditis caused by an organism resembling Corynebacterium striatum; Markowitz SM et al.; An organism resembling Corynebacterium striatum was isolated from the blood of a patient with acute aortic valvular insufficiency and no history of valvular heart disease . At autopsy, histopathologic examination of the aortic valve revealed pleomorphic gram-positive bacilli and destruction of valvular tissue . Our isolate differed from other nondiphtherial corynebacteria, including the type strain of C . striatum (ATCC 6940), in its ability to reduce nitrite . Nitrite reduction may be useful for distinguishing strains of corynebacteria. Arteriosclerosis, 1990 Jan-Feb, 10(1), 8 - 16 Effects of activation on lipid and lipoprotein metabolism in murine macrophages; Kraemer FB et al.; The effects of activation on lipid and lipoprotein metabolism were examined in resident murine macrophages, inflammatory cells elicited by thioglycolate, primed cells elicited by pyran copolymer, and activated cells elicited by Corynebacterium parvum . Low density lipoprotein receptors were reduced by 70%, while scavenger receptors were reduced 60% in activated cells . Basal cholesteryl ester and triglyceride synthesis were increased fourfold in activated cells, whereas the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase was high in resident cells and progressively declined by greater than 80% in activated cells . Activities of neutral cholesteryl esterase and neutral triglyceride lipase were increased two- to fourfold in inflammatory, primed, and activated macrophages . These results demonstrate the diverse changes in lipid and lipoprotein metabolism that occur with activation and emphasize how the behavior of macrophages in atherosclerotic lesions can be altered by activation. Arch Ophthalmol, 1990 Jan, 108(1), 107 - 12 Pathogenicity of Acanthamoeba and a Corynebacterium in the rat cornea; Badenoch PR et al.; Acanthamoeba keratitis is a sight-threatening disease that is difficult to treat . The development of an animal model is necessary for many of the studies required to improve visual outcome in human patients . A rat model is proposed that is dependent on coinoculation of amebae and corynebacteria into the corneal stroma . The infective dose was determined for a virulent Acanthamoeba isolate and was used to screen 17 other isolates, including 7 from the human cornea . A total of 6 were infective in the rat cornea . The model should be useful for controlled in vivo studies of this intractable condition. Mikrobiyol Bul, 1990 Jan, 24(1), 79 - 87 {Immunotoxins}; Gedikoglu S; An immunologic approach besides surgical, radiotherapeutic, chemotherapeutic methods in the treatment of cancer was proposed for the first time by Paul Ehrlich in 1906 . The structure, produced by conjugating the antibodies against the tumor cells with a toxin molecule by Moolten and Cooperband (1970) was named as Immunotoxins . Producing antibodies with the desired amount and purity was achieved by improving the Hybridoma technology, Immunotoxins were tried to be produced by conjugating the monoclonal antibodies with the different toxic agents . Removing the receptor binding region of the toxin molecule was demonstrated to be more effective although good results were obtained from the studies utilising the toxin of Corynebacterium diphtheriae . Toxin production with the desired amount and structure was accomplished by the improved genetic methods . Although in vivo applications of the immunotoxins are still limited, it is believed that they will be important therapeutic agents in the future. Acta Microbiol Hung, 1990, 37(2), 201 - 6 Revision of the validity of CAMP tests for Listeria identification . Proposal of an alternative method for the determination of haemolytic activity by Listeria strains; Vazquez-Boland JA et al.; The validity of CAMP tests with Staphylococcus aureus and Rhodococcus (Corynebacterium) equi as defined for Listeria identification was revised . This characterization method appeared to be unreliable for two reasons: first, a positive CAMP test with R . equi is not specific for Listeria ivanovii as Listeria monocytogenes (and Listeria seeligeri) give also a clear positive reaction; second, doubtful reactions could be observed with S . aureus when assaying haemolytic and non-haemolytic Listeria strains (possibility of false negative and false positive results; subjectivity of the interpretation) . The use of a Microplate technique previously described instead of CAMP tests is proposed for the reliable demonstration of the haemolytic character of Listeria in the routine identification of these organisms. Immunopharmacol Immunotoxicol, 1990, 12(3), 457 - 77 Augmentation of host defense against bacterial infection pretreated intraperitoneally with an alpha-glucan RBS in mice; Takeda Y et al.; Protection against Listeria monocytogenes and Escherichia coli in mice was enhanced by an intraperitoneal (i.p.) administration of polysaccharide "RBS" . Peritoneal macrophages from mice administered i.p . with 30 mg/kg doses of RBS 4 days earlier exhibited increased scavenger functions as assessed by in vivo phagocytosis, in vitro intracellular killing and generation of superoxide anion . When cytokine production of the macrophages was assessed by biological assay and Northern blotting analysis, interleukin (IL)-1 production and IL-1 alpha gene expression were significantly increased in macrophages from RBS-treated mice . On the other hand, tumor necrosis factor (TNF) alpha gene was expressed in macrophages from RBS-treated mice at a much reduced level as compared with those in mice treated i.p . with Corynebacterium parvum on 4 days earlier . In correlation with expression of TNF gene in the macrophages, RBS-treated mice were less susceptible to the lethal toxicity of LPS than C.parvum-treated mice . In RBS-treated mice, in vivo elimination of bacteria was enhanced at the early phase of infection with L.monocytogenes or E.coli, resulting in augmentation of host defense against these bacterial infection . These results suggest that adequately enhanced activities of macrophages acting as scavenger phagocytes play important roles in the enhanced resistance against bacteria in mice treated i.p . with RBS. Semin Surg Oncol, 1990, 6(6), 364 - 8 Immunotherapy of gynecologic malignancies; Foon KA et al.; Systemic Corynebacterium parvum and BCG have limited activity in gynecologic malignancies . Although intraperitoneal C . parvum is active, its toxicity is prohibitive . Intraperitoneal alpha-interferon is an active second line agent for minimal residual disease following combination chemotherapy . Intraperitoneal interferon trials are ongoing . Alpha-interferon is also active against lower genital tract condyloma acuminata . Sufficient numbers of patients have not been evaluated to determine the activity of interleukin-2(IL-2) in gynecologic malignancies . Radioisotope labeled monoclonal antibodies can image gynecologic malignancies and may have a future therapeutic role . The last decade has witnessed a substantial growth in immunotherapy and has demonstrated a role for biologic agents in cancer therapy . Continued improvement in biologic therapies should lead to major advances in gynecologic cancer diagnosis and therapy. J Environ Pathol Toxicol Oncol, 1990 Jan-Apr, 10(1-2), 41 - 4 Antimicrobial action of roquefortine; Kopp-Holtwiesche B et al.; Roquefortine, a secondary metabolite of Penicillium roqueforti, was investigated with regard to its effect on microorganisms . The growth of gram-positive bacteria was inhibited by roquefortine, whereas gram-negative bacteria were not influenced at all . The minimal inhibitory concentrations for gram-positive organisms were about 80 micrograms/mL cells on the average . The sensitive bacterium Corynebacterium flaccumfaciens was chosen for characterizing the antimicrobial action of the mycotoxin . Addition of low roquefortine concentrations at the beginning of bacterial growth resulted in prolongation of both the lag and logarithmic phases . High toxin concentrations (100 micrograms/mL cells) added to growing cells caused prolongation of logarithmic growth and decreased maximum cell density . This effect could be abolished when the cells, inhibited during the logarithmic growth, were centrifuged, washed, and transferred into fresh nutrient medium . In that case a second prolonged lag phase occurred, but the following logarithmic growth was comparable to normal cell growth . These results indicate that the antimicrobial effect of roquefortine is bacteriostatic but not bactericide . The growth inhibition might be a consequence of a roquefortine influence on bacterial respiration . The uptake of oxygen was reduced to 50% at a toxin concentration of 100 micrograms/mL cells. Graefes Arch Clin Exp Ophthalmol, 1990, 228(5), 458 - 60 External eye flora as a nutrient source for Acanthamoeba; Larkin DF et al.; Certain bacteria cause excystment of Acanthamoeba from cyst to trophozoite form and are then ingested by migrating trophozoites . We studied the response of Acanthamoeba cysts to inoculation on agar seeded with three types of commensal eye bacteria and Escherichia coli . Amoebae excysted on all bacteria tested, and the migration rate of Acanthamoeba trophozoites on each was compared . Acanthamoeba migrated with equal speed on E . coli and Staphylococcus epidermidis . Migration was observed, but was more slow on Micrococcus and Corynebacterium . Commensal bacteria on the eyelids, conjunctiva and tear film may have a role in pathogenesis of Acanthamoeba keratitis. Rev Elev Med Vet Pays Trop, 1990, 42(4), 512 - 4 Prevalence of mastitis in imported Friesian cows in Sudan; Abdelrahim AI et al.; Three hundred twenty-two lactating Friesian cows were examined for mastitis by different diagnostic techniques . The predominant pathogens encountered were Staphylococci, Streptococci, Corynebacterium and Escherichia coli spp. Med Lab Sci, 1990 Jan, 47(1), 36 - 41 Corynebacteria: incidence among samples submitted to a clinical laboratory for culture; Marshall RJ et al.; Over the period of one year, 83 corynebacteria isolates were identified in our laboratory, and their clinical relevance assessed by reference to patients whose clinical notes were available . Eleven species of corynebacteria were identified including four biotypes of C . jeikeium; six organisms were non-typeable; C . jeikeium and C . xerosis predominated . Species identified in the literature as causing clinical infection were also isolated--though in smaller proportions--as were strains of C . jeikeium which were not multi-resistant to antibiotics . Immuno-compromised patients and those with renal impairment had an increased frequency of corynebacteria . The isolation of C . jeikeium from the blood of a neonate suggests that this may be a potential pathogen in these patients . Antibiotic susceptibility of an organism was not a reliable marker of significance, and a reliable biotyping scheme should be adopted. Immunopharmacol Immunotoxicol, 1990, 12(3), 363 - 87 Characterization of Corynebacterium parvum-induced suppressor cells of mouse NK and ADCC activity; Santoni A et al.; Treatment of mice with Corynebacterium parvum (Cp) resulted in a substantial decrease in natural killer activity in the spleen at 10 days . The decrease in cytotoxicity was associated with the presence of splenic nonadherent (NA) suppressor cells, capable of inhibiting natural as well as antibody-dependent cellular cytotoxicity (ADCC) . The nonadherent suppressor cells appeared to be null cells, lacking detectable expression of Thy 1, L3T4 (CD4), Lyt 2 (CD8), or asialo-GM1 and could be physically separated from cells with NK activity by centrifugation on Percoll discontinuous density gradients . Our results indicate that Cp can negatively modulate cytolytic functions of NK cells by inhibiting the effector phase of cytotoxicity. Cancer Immunol Immunother, 1990, 31(2), 121 - 7 Cyclophosphamide and abrogation of tumor-induced suppressor T cell activity; Hoover SK et al.; Previously we have demonstrated that the in vitro generation of P815-specific anti-tumor cytotoxic T lymphocytes (CTL) was suppressed by splenic suppressor T cells from late tumor-bearing hosts (TBH) . Suppression is not caused by in vitro growth of P815 from splenic metastases, since suppression was also seen with spleen cells from late TBH mice bearing a hypoxanthine/aminopterin/thymidine-sensitive subline (PHS-5) of P815 in the presence of HAT . Cyclophosphamide has been shown to inhibit the induction of suppressor cells selectively in a number of immune responses, but evidence that it can inhibit active tumor-induced suppressor T cells is limited . We have found that suppressor T cells already induced by P815 in syngeneic late TBH are sensitive to low doses of cyclophosphamide (50 mg/kg) given 1 day before spleen harvest, but the in vitro CTL response of late TBH spleen cells could not be restored by pretreating the mice with cyclophosphamide, even when exogenous interleukin-2 was added to the cultures . Although 50 mg/kg cyclophosphamide did not inhibit the CTL response of spleen cells from mice immunized with P815 + Corynebacterium parvum, the same dose of cyclophosphamide eliminated the CTL response of spleen cells from early TBH . Interleukin-2 (IL-2) did not overcome this effect of cyclophosphamide, suggesting a direct effect on CTL . "Ultra-low" -dose cyclophosphamide (10 mg/kg) did not adversely effect early TBH CTL but was still able to eliminate suppressor T cell activity from late TBH . Nevertheless, late TBH CTL remained unresponsive after pretreatment of mice with ultra-low-dose cyclophosphamide, even when exogenous IL-2 was added in vitro . CTL precursor frequency analyses demonstrated that cyclophosphamide pretreatment had little or no effect on the numbers of CTL precursors from early TBH . Late TBH CTL precursor cells were not detectable in these studies, with or without suppressor T cell inhibition by cyclophosphamide pretreatment . Thus, it appears that most CTL precursor cells may be lost or irretrievably inactivated in the spleens of late TBH mice. Immunology, 1990 Jan, 69(1), 127 - 33 Relation between locomotion, chemotaxis and clustering of immune cells; Wilkinson PC; Experiments were designed to discover whether locomotor or chemotactic events are needed for clustering of lymphocytes with accessory cells or, conversely, whether clustering precedes the activation of lymphocyte locomotion . The time-courses of clustering and locomotor activation were compared and the behaviour of moving cells during cluster formation was filmed . Human lymphocytes direct from blood were activated by culture for 24-48 hr with anti-CD3 antibody or in allogeneic mixed leucocyte reactions (AMLR) . The proportion of clustered and locomotor lymphocytes was low at the beginning of culture . Clusters appeared during the first few hours, before the increase in numbers of locomotor lymphocytes . Filming gave no evidence that the cells attracted one another chemotactically to form clusters . Rather, cells made chance contact by random locomotion and then remained adherent, though lymphocytes very close (less than or equal to 10 microns) to clusters did show increased pseudopod formation towards the cluster . However, the behaviour of motile lymphocytes responding to monocytes or macrophages given a phagocytic stimulus was different . Human monocytes which ingested opsonized zymosan released a material during but not following phagocytosis which caused an immediate increase in polar shape-change in lymphocytes . Macrophages from Corynebacterium parvum-induced mouse peritoneal exudates, given a phagocytic stimulus (opsonized Candida albicans), acted as sources of chemotactic gradients which attracted nearby lymphocytes to form clusters . This was due to brief release of a material immediately following phagocytosis, but after 15 min or so the macrophages no longer attracted nearby cells . These experiments suggest that, during induction of an immune response to a non-phagocytic stimulus, clusters form slowly by random contact followed by preferential adhesion . However, after phagocytosis, there may be a chemotactic response to the ingesting macrophage . This may help to focus lymphocytes onto macrophages which present microbial antigens. Eur J Clin Microbiol Infect Dis, 1990 Jan, 9(1), 30 - 3 Comparative in vitro activity of the new erythromycin derivative dirithromycin against gram-positive bacteria isolated from cancer patients; Rolston KV et al.; The in vitro activity of dirithromycin (LY-237216), a new macrolide erythromycin derivative, was compared to that of four other agents (clarithromycin, erythromycin, roxithromycin, clindamycin) against 334 gram-positive isolates obtained from cancer patients . Dirithromycin was similar in potency and antimicrobial spectrum to the other agents tested . It was very active against beta-haemolytic streptococci and Streptococcus pneumoniae, and moderately active against penicillin and methicillin susceptible Staphylococcus aureus, Bacillus spp., Listeria monocytogenes and Corynebacterium jeikeium . Erythromycin resistant organisms were also resistant to dirithromycin. Pol Arch Weter, 1990, 30(3-4), 155 - 61 {Changes in the aortic arch of calves with bacterial infections}; Balbierz H et al.; The autopsy and histopathological changes in the wall of aortic arch were presented in calves died of pulmonary and intestinal disorders . The bacteriological examinations of the internal organs demonstrated: E . coli, Pseudomonas aeruginosa Corynebacterium et al . Gelatinous edematous++ and hemorrhagic inflammatory centers of aortic arch adventitia were distinctly visible during thorax examination . In aortic arch the diagonal fissures of endothelium and deeper layers of the wall were found . In microscopical examination the heavy degenerative changes in the arch endothelium and deeper layers were observed . The fissures were from 0.5 to several millimeters in length and penetrated 1/3 to 1/2 thickness at the wall . The authors suggested that starting point of this changes are Gram-negative bacillary inflammatory centers in lungs . The bacterial endotoxins and vasoactive substances liberated by altered cells penetrated the aortic arch by the tissues continuity and in contact way. Chemotherapy, 1990, 36(6), 403 - 6 In vitro activity of fleroxacin against aerobic gram-positive bacteria including Corynebacterium jeikeium; Hohl P et al.; Fleroxacin (Ro 23-6240, AM-833), a new fluoro-4-quinolone, was tested in vitro against 273 gram-positive clinical isolates . Norfloxacin, a quinolone mostly used in urinary tract infections, was included as a standard . Overall, in vitro activities of fleroxacin and norfloxacin were superposable, but interestingly, fleroxacin was two- to fourfold more active against Staphylococcus aureus, and 1 mg/l of fleroxacin inhibited all 5 multiply resistant Corynebacterium jeikeium tested . While both quinolones were similarly active weight by weight, fleroxacin with its more favorable pharmacokinetic properties harbors the added promise of clinical usefulness in systemic infections against susceptible isolates. Vopr Med Khim, 1990 Jan-Feb, 36(1), 27 - 31 {Deoxyribonuclease from Corynebacterium diphtheriae: dynamics of synthesis and properties}; Iusupova DV et al.; Diphtheritic bacteria of PW-8 Massachusetts strain produced into cultural medium only one nucleotidase--endoDNAase . The enzyme was synthesized by the cells during the exponential phase of growth . The DNAase was purified 500-fold and exhibited properties specific to neutral-alkaline DNAases (pH optimum about 7.5, absolute requirements for Me2+, single-step mechanism of substrate hydrolysis) . The following properties were typical for the enzyme: absence of distinct specificity to structure of bases surrounding the hydrolyzed bond, formation of 5'-end phosphate groups and slightly higher preference to denatured DNA. FEMS Microbiol Lett, 1990 Jan 1, 54(1-3), 299 - 301 DNA sequence homology between attB-related sites of Corynebacterium diphtheriae, Corynebacterium ulcerans, Corynebacterium glutamicum, and the attP site of gamma-corynephage; Cianciotto N et al.; Chromosomal restriction fragments of Corynebacterium ulcerans and C . diphtheriae, containing an integration site for corynephages of the beta family, show homology on Southern blots . Homologous DNA in also found in the soil isolate C . glutamicum, although this strain is not susceptible to beta-corynephages . Three of these DNA fragments, one for each bacterial strain, and a fragment of gamma-corynephage DNA previously shown to contain the phage integration site, were cloned and sequenced . Alignment of the 3 bacterial sequences shows a very high degree of homology in a stretch of ca 120 nucleotides, whereas the rest of the sequences is generally non-homologous . Within this common bacterial portion, a segment of ca . 96 nucleotides (core sequence) is also highly homologous to the phage sequence . The first half (ca . 50 bp) of the core sequence is identical in all aligned sequences whereas the second half, which is largely occupied by a stem-and-loop structure, contains point mutations peculiar to each clone . The described sequences are likely to be involved in phage integration/excision processes. FEMS Microbiol Lett, 1990 Jan 1, 54(1-3), 119 - 23 Localization of an origin of replication in Corynebacterium diphtheriae broad host range plasmid pNG2 that also functions in Escherichia coli; Serwold-Davis TM et al.; Subcloning and protoplast transformation studies identified a 2.6 kb fragment of Corynebacterium diphtheriae plasmid pNG2 which contains an origin of replication (oriR) . Molecular combination of the 2.6 kb oriR cartridge with Escherichia coli plasmid pUC18CmR enabled the E . coli cloning vector to replicate within several species of Corynebacterium host cells . A 2.6 kb plasmid formed from the oriR cartridge alone is capable of replicating in E . coli . This suggests that a single origin could be used in vectors shuttling between Corynebacterium spp . and E . coli. Acta Vet Hung, 1990, 38(4), 231 - 8 Survey of Actinobacillus (Haemophilus) pleuropneumoniae infection in swine by different methods; Molnar E; Lung and serum samples from pigs that died or were emergency-slaughtered in a pooled, conventional fattening herd were examined to survey Actinobacillus pleuro-pneumoniae infection and to compare the sensitivity of different testing methods . A total of 110 lungs were used for cultural isolation of the agent and direct immunofluorescence (IF) of impression smears . Boiled lung suspensions were tested by coagglutination (Co-A) and agar gel precipitation (AGP) . Eighty-seven sera were tested along with lung samples from the same pigs . The lungs yielded a varied bacterial flora most often containing Pasteurella multocida and less frequently Actinomyces (Corynebacterium) pyogenes, E . coli and Salmonella . A . pleuropneumoniae was isolated from 30 lungs: from 22 lungs it grew out in pure culture, from 7 as mixed culture with P . multocida and from 1 as mixed culture with A . pyogenes . The number of positive samples obtained by the different methods was as follows: coagglutination test (with boiled lung suspensions): 63 (57.3%); immunofluorescence: 43 (39.2%); AGP test (with serum): 31 (35.6%); AFP test (with boiled lung suspension): 25 (22.7%) . A total of 23 samples (20.7%) were negative by all serological tests and by cultural isolation . Most samples gave positive results by two or more tests while 26 samples only by one test (most often, on 13 occasions, by the Co-A test) . The Co-A test detected antigenic components of serotypes that have not been isolated in Hungary so far . This indicates that it is not enough to test one strain from a given lung sample: several colonies must be cultured and serotyped.(ABSTRACT TRUNCATED AT 250 WORDS) Nat Immun Cell Growth Regul, 1990, 9(6), 387 - 96 Nonspecific stimulation of host defense by Corynebacterium kutscheri . II . Isolation of the active moiety; Kita E et al.; The isolation and determination of biological activities of the active component of Corynebacterium kutscheri were attempted in the present investigation . The antitumor effect was confined to the subcellular particle fraction of this bacterium and was associated with a molecule of glycoprotein nature (40,000-38,000 Daltons) isolated from this fraction by affinity chromatography with concanavalin A-Sepharose 4B . This substance exerted mitogenic activity on C3H/HeJ splenocytes and T cells, stimulatory activity on macrophages, and further exhibited antitumor effect on P388 leukemia in CDF1 mice . The Winn assay disclosed that the antitumor effect induced by this substance was dependent on L3T4+ T cells . Furthermore, both the mitogenic and antitumor activity of this moiety were resistant to heating at 100 degrees C for 30 min or RNase digestion, but sensitive to trypsin digestion, or low or high pH . These results indicate that the antitumor effect of C . kutscheri is attributable to the heat-stable glycoprotein moiety which can directly stimulate T cells and macrophages. Med Dosw Mikrobiol, 1990, 42(3-4), 178 - 83 {Milk chloride level as an indicator of bovine mastitis}; Sender G et al.; The aim of this study was to determine an influence of udder infection on milk chloride level and on milk productivity of cows of black and white race . Bacteriological analysis was performed by bacterial isolation from milk collected in sterile conditions from single lobes of mammary gland . The study was aimed to detect the following bacteria: Streptococcus agalactiae, Streptococcus uberis, Streptococcus agalactiae, Staphylococcus aureus, Staphylococcus epidermidis, Actinomyces pyogenes, Enterobacteriaceae rods, Corynebacterium bovis, and Micrococcus sp . Milk chloride level was determined by burette method in 1250 milk samples collected from entire udder . Milk productivity was determined on the day of bacterial isolation . Statistical analysis of the results of the study on udder infection, milk chloride level, and milk productivity of mammary gland did reveal lack of a simple correlations between those parameters and in the indirect manner indicated an influence of inappropriate maintenance conditions of tested cows on the health condition of their udder . It seems possible that alkalosis and acidosis in cows, taken into consideration in discussion section, could constitute a factor influencing the frequency of mastitis incidence. Appl Microbiol Biotechnol, 1990 Jan, 32(4), 443 - 8 Cloning of a DNA fragment from Corynebacterium glutamicum conferring aminoethyl cysteine resistance and feedback resistance to aspartokinase; Thierbach G et al.; The Corynebacterium glutamicum/Escherichia coli shuttle vector plasmid pZ1 was used to clone the S-(2-aminoethyl)-D,L-cysteine (AEC)-resistance gene from a lysine-excreting, AEC-resistant strain of C . glutamicum, the aspartokinase activity of which was released from feedback inhibition by mixtures of lysine and threonine or AEC and threonine respectively . A recombinant plasmid designated pCS2 carrying a 9.9-kb chromosomal insert that conferred AEC resistance and the ability to excrete lysine to its host was isolated . The aspartokinase activity of the pCS2-carrying strain was resistant towards inhibition by mixtures of lysine and threonine or AEC and threonine respectively . By deletion analysis the DNA region conferring AEC resistance to the host and feedback resistance to its aspartokinase activity could be confined to a 1.2-kb DNA fragment. J Biol Chem, 1989 Dec 5, 264(34), 20438 - 47 Glutaredoxin from rabbit bone marrow . Purification, characterization, and amino acid sequence determined by tandem mass spectrometry; Hopper S et al.; A glutaredoxin was purified from rabbit bone marrow, and its amino acid sequence was determined by high performance tandem mass spectrometry . The sequences of peptides generated by digestion with trypsin alone or in combination with thermolysin were determined from their collision-induced dissociation (CID) mass spectra . Alignment of these sequences and additional sequence information were obtained from the collision-induced dissociation mass spectra of peptides obtained from digestion of the intact protein with Staphylococcus aureus V8 protease and alpha-chymotrypsin . The resulting sequence of 106 amino acids is as follows: Ac-Ala-Gln-Glu-Phe-Val-Asn-Ser-Lys-Ile-Gln-Pro-Gly-Lys-Val-Val-Val-Phe- Ile-Lys-Pro-Thr-Cys-Pro-Tyr-Cys-Arg-Lys-Thr-Gln-Glu-Ile-Leu-Ser-Glu-Leu- Pro-Phe - Lys-Gln-Gly-Leu-Leu-Glu-Phe- Val-Asp-Ile-Thr-Ala-Thr-Ser-Asp-Met-Ser-Glu-Ile- Gln-Asp-Tyr-Leu-Gln-Gln-Leu-Thr-Gly-Ala-Arg- Thr-Val-Pro-Arg-Val-Phe-Leu-Gly-Lys-Asp-Cys-Ile- Gly-Gly-Cys-Ser-Asp-Leu-Ile-Ala-Met-Gln-Glu-Lys- Gly-Glu-Leu-Leu-Ala-Arg-Leu-Lys-Glu-Met-Gly- Ala-Leu-Arg-Gln . This glutaredoxin strongly resembles the corresponding calf and pig proteins (known as glutaredoxin and thioltransferase, respectively) with respect to its primary structure and enzymatic activity as a GSH:disulfide thioltransferase, an activity also found for the glutaredoxin from Escherichia coli . However, rabbit glutaredoxin was not active as a hydrogen donor for the reduction of ribonucleotides in the presence of the ribonucleotide reductases from rabbit bone marrow, Lactobacillus leichmannii, and Corynebacterium nephridii. FEMS Microbiol Lett, 1989 Dec, 53(3), 299 - 303 High efficiency electroporation of intact Corynebacterium glutamicum cells; Liebl W et al.; High-frequency electroporation of whole Corynebacterium glutamicum cells without enzymatic pretreatment was achieved . Under optimized conditions concerning growth stage, washing of cells, cell concentration and pulse parameter transformation efficiencies of far more than 10(7) transformants per microgram pWST4B plasmid DNA were reached . Using electroporation, linearised and subsequently religated plasmid as well as chimeric ligase reaction products were directly introduced into C . glutamicum with reasonable efficiencies . Electrotransformation efficiency was reduced about 10(5)-fold for plasmid DNA cycled through E . coli JM83 . Restriction deficient mutants of C . glutamicum were isolated which could be efficiently transformed with foreign DNA. Immunology, 1989 Dec, 68(4), 520 - 5 Systemic administration of IL-2 induces lymphokine-activated killer (LAK) cells capable of killing macrophages in various tissues; Kamitani T et al.; Murine lymphokine-activated killer (LAK) cells induced by systemic high-dose recombinant human interleukin 2 (IL-2) administration lysed fresh syngeneic peritoneal macrophages (M phi) . LAK cells lysed resident peritoneal M phi and M phi activated in vivo with thioglycollate (TG), Corynebacterium parvum (C . parvum), or Bacillus Calmette-Guerin (BCG) . The induction of anti-M phi cytolytic activity was seen in the spleen, liver, lung, lymph nodes and peritoneal cavity, but was not observed in the thymus . Fluorescence analysis revealed that the majority of infiltrated cells in the peritoneal cavity of IL-2-administered mice were Thy-1+, asialo GM1+, L3T4-, Ly2- . Surface marker analysis on peritoneal exudate cells (PEC) from IL-2-administered mice with depletion techniques using antibody (Ab) and complement (C) indicated that Thy-1+, asialo GM1+, L3T4-, Ly 2- cells were responsible for anti-M phi lysis . These studies indicate that the in vivo administration of IL-2 induces LAK cells capable of killing M phi in various tissues. Nippon Juigaku Zasshi, 1989 Dec, 51(6), 1173 - 8 Studies on route of immunization with a mixture of killed parasites and adjuvants against Babesia rodhaini infection in mice; Saeki H et al.; A series of experiments were undertaken to determine the most effective route of immunization with a mixture of killed Babesia rodhaini antigen (S antigen) and formalin-fixed Corynebacterium parvum (Propionibacterium acnes) bacterin (CPB) against challenge infection with B . rodhaini 3 weeks later . The mice pretreated with S antigen and CPB mixture intraperitoneally, but not intramuscularly, were significantly resistant to intraperitoneal (IP) or intravenous (IV) challenge with 10(6) organisms . The survival rates were 70.0 (IP challenge) and 60.0% (IV challenge) respectively . Fairly protective activities were equally produced in mice intravenously pretreated with S antigen and CPB with survival rates of 60.0% against IV challenge, but 30% against IP . These results indicated that the IP injection of S antigen and CPB mixture is desirable route for immunization against subsequent IP or IV challenge with B . rodhaini . On the other hand, lower protective effect was reconfirmed in the mice treated with S antigen and Freund's Complete adjuvant, regardless of immunization routes in the additional experiment . The survival rates were 33.3, 14.3 and 11.8% in the intraperitoneally, intramuscularly and subcutaneously-treated mice respectively against IP challenge with 10(6) organisms. Gynecol Oncol, 1989 Dec, 35(3), 378 - 82 Second-look laparotomy in the patient with minimal residual stage III ovarian cancer (a Gynecologic Oncology Group Study); Creasman WT et al.; One hundred eighty-six patients with minimal residual Stage III ovarian cancer (tumor mass less than or equal to 3 cm) were treated in a prospective randomized protocol (melphalan with or without Corynebacterium parvum) . As per protocol 84 patients were eligible and underwent a second-look laparotomy with 41 (49%) having negative findings for persistent malignancy . Factors which affected survival after second look were presence or absence of macroscopic disease, age, and grade . Depending upon these prognostic factors, survival at 4 years after second look ranged from 31 to 100% . The role of second-look laparotomy is examined relative to these results. J Clin Microbiol, 1989 Dec, 27(12), 2869 - 70 Corynebacterium xerosis as a cause of vertebral osteomyelitis; Krish G et al.; We report a patient who developed Corynebacterium xerosis vertebral osteomyelitis 6 months following a decompressive laminectomy . Prolonged parenteral and subsequent oral therapy for 11 months resulted in apparent cure . This is the first reported case of vertebral osteomyelitis caused by C . xerosis. J Clin Microbiol, 1989 Dec, 27(12), 2736 - 43 Comparison of the nasal bacterial floras in two groups of healthy subjects and in patients with acute maxillary sinusitis; Jousimies-Somer HR et al.; The nasal bacterial flora was studied in 183 healthy men entering military service (entry group), 103 healthy recruits in service (service group), and 185 recruits with acute maxillary sinusitis . The 267 nasal and ipsilateral sinus aspirate findings in the same patients with acute maxillary sinusitis were compared pairwise . In the entry group presumed sinus pathogens were only rarely isolated from the nasal cavities: Haemophilus influenzae in 4%, Streptococcus pneumoniae in 1%, Branhamella catarrhalis in 3%, and Streptococcus pyogenes in 0% . The corresponding isolation frequencies in the service group were 19, 13, 3, and less than 1%, respectively, and those in the group with acute maxillary sinusitis were 61, 25, 7, and 6%, respectively . Suppression of the major components of the normal nasal flora, Corynebacterium sp., coagulase-negative staphylococci, Propionibacterium acnes, and Staphylococcus aureus, was seen in the group with acute maxillary sinusitis and also occasionally in the service group . When a sinus aspirate culture yielded a presumed sinus pathogen, the same pathogen was found in the nasal samples in 91% of the cases . The predictive value of a pathogen-positive nasal finding was highest (93.8%) for S . pyogenes, followed by 77.7% for H . influenzae and 68.7% for S . pneumoniae, and lowest (20%) for B . catarrhalis. J Immunol, 1989 Dec 1, 143(11), 3641 - 6 Regulation of macrophage physiology by L-arginine: role of the oxidative L-arginine deiminase pathway; Albina JE et al.; The L-arginine content of the extracellular fluid in sites of predominant macrophage infiltration is reduced below plasma levels due to the activity of macrophage-derived arginase . Investigation of the effects of altered L-arginine availability on macrophage physiology reveals that culture of rat peritoneal macrophages in media containing L-arginine in the concentrations present in inflammatory lesions (less than 0.1 mM) enhances activation-associated functions . In contrast, culture in the higher L-arginine concentrations found in standard tissue culture media (0.4 to 1.2 mM) suppresses most macrophage functions (superoxide production, phagocytosis, and protein synthesis) . An exception is the tumor cytotoxicity of Corynebacterium parvum-elicited macrophages which is enhanced by culture in supraphysiologic concentrations of L-arginine . Work reported here investigated the mechanisms for these L-arginine-dependent effects and, more specifically, the role of the recently described oxidative L-arginine deiminase pathway in the regulation of macrophage physiology . Overnight culture of resident or C . parvum-elicited peritoneal macrophages in media containing increasing concentrations of L-arginine (6 microM to 1 mM) resulted in: inhibition of electron transport chain activity (resident and C . parvum-elicited macrophages), increased lactate production (resident macrophages), and decreased ATP content (resident and C . parvum-elicited macrophages) . In line with these findings, viability was markedly decreased after 2 days of culture when the initial L-arginine concentration was greater than or equal to 0.1 mM . As shown before, increasing media concentrations of L-arginine were associated with suppression of superoxide production and cytotoxicity in resident macrophages, and with reduced superoxide production and increased cytotoxicity in C . parvum-elicited macrophages . All L-arginine-dependent metabolic a