Corynebacter

Nihon Kyobu Shikkan Gakkai Zasshi, 1996 Aug, 34(8), 864 - 9
{Roles of inflammatory cells in the pathogenesis of acute lung injury in guinea pigs exposed to heat-killed bacteria}; Tasaka S et al.; To study the contribution of polymorphonuclear (PMN) and mononuclear (MN) phagocytes to the development of acute lung injury, we studied lung injury after intratracheal instillation of lipopolysaccharide (0.02 mg/kg) in guinea pigs previously exposed to heat-killed Corynebacterium parvum . In on group, cyclophosphamide was given to deplete peripheral PMNs . In another group, gadolinium chloride (GdCl3) was injected to suppress the function of MNs . Four hours after instillation of lippoly soccharide, the animals were killed, bronchoalveolar lavage was done, and the lungs were examined histopathologically . 125I-labeled albumin was injected to estimate the endothelial damage, and 131I-labeled albumin was injected to correct for blood contamination in the samples . In the group given cyclophosphamide, lung injury was no less than in the control group . In contrast, lung injury was less sever in the group given GdCl3 than in the control group . These findings suggest that MN are important in the pathogenesis of lung injury, especially in individuals who are immunologically primed by infection.

Eur J Clin Microbiol Infect Dis, 1996 Aug, 15(8), 657 - 62
Species identities and antimicrobial susceptibilities of corynebacteria isolated from various clinical sources; Riegel P et al.; Over a 14-month period, 415 clinical isolates of coryneform gram-positive rods were recovered from various sources and identified to the species level according to recent identification schemes . Corynebacterium urealyticum, Corynebacterium striatum, Corynebacterium amycolatum, and Corynebacterium jeikeium predominated, accounting for 63% of all isolates . Corynebacterium accolens, Corynebacterium striatum, Corynebacterium argentoratense, Corynebacterium propinquum and Corynebacterium pseudodiphtheriticum were mostly recovered from the respiratory tract, whereas Corynebacterium afermentans, CDC group G, and Corynebacterium jeikeium were mainly isolated from blood . None of the isolates was identified as Corynebacterium diphtheriae or Corynebacterium xerosis . Ampicillin resistance was detected in Corynebacterium jeikeium (96%) and Corynebacterium urealyticum (99%) and varied among Corynebacterium amycolatum (56%) and CDC group G (26%) . These data emphasize the need for an accurate identification of coryneform organisms at the species level and for antimicrobial susceptibility testing of these organisms.

J Radiol, 1996 Aug, 77(8), 571 - 3
{Parietal calcifications of the kidney pelvis in Corynebacterium urealyticum urinary infection}; Maciejewski C et al.; Linear renal pelvis calcifications in a native kidney due to Corynebacterium urealyticum are described . These micro-organisms have an opportunistic behaviour and can be responsible for nosocomial urinary tract infection . Alkaline incrusted cystitis with linear vesical calcifications are considered as the most typical pattern . Renal pelvis may also be concerned, with parietal calcifications.

Proteins, 1996 Aug, 25(4), 514 - 6
Expression, purification, and crystallization of meso-diaminopimelate dehydrogenase from Corynebacterium glutamicum; Reddy SG et al.; The gene encoding the meso-diaminopimelate dehydrogenase (DAPDH) from Corynebacterium glutamicum was over-expressed and purified to homogeneity . Crystals of the binary DAPDH-NADP+ complex were obtained from solutions of polyethylene glycol 8000, 100 mM sodium cacodylate, pH 6.5, and 150-300 mM Mg(OAc)2 . The crystals diffract to 2.2 A, belong to the orthorhombic space group P2(1), and contain two molecules per asymmetric unit.

Arch Microbiol, 1996 Aug, 166(2), 76 - 82
A Corynebacterium glutamicum gene encoding a two-domain protein similar to biotin carboxylases and biotin-carboxyl-carrier proteins; Jager W et al.; Following the analysis of transposon Tn5432-induced mutants of Corynebacterium glutamicum ATCC 13032, a gene encoding a protein with a biotin-binding motif was cloned . The DNA sequence of this gene revealed an open reading frame encoding 591 amino acids with a calculated mol . mass of 63.4 kDa . The protein is composed of two domains, an N-terminal biotin carboxylase and a C-terminal biotin-carboxyl-carrier protein, that are highly similar to corresponding subunits from prokaryotic and eukaryotic biotin enzymes . Over 70% identity was found to a protein from Mycobacterium leprae proposed to be part of an acyl-CoA carboxylase . Since it was not possible to inactivate the C . glutamicum gene, the gene most likely encodes a subunit of the essential acetyl-CoA carboxylase, which catalyzes the committed step in fatty acid synthesis.

J Bacteriol, 1996 Aug, 178(15), 4412 - 9
Mutations in the Corynebacterium glutamicum proline biosynthetic pathway: a natural bypass of th proA step; Ankri S et al.; Two chromosomal loci containing the Corynebacterium glutamicum ATCC 17965 proB and proC genes were isolated by complementation of Escherichia coli proB and proC auxotrophic mutants . Together with a proA gene described earlier, these new genes describe the major C . glutamicum proline biosynthetic pathway . The proB and proA genes, closely linked in most bacteria, are in C . glutamicum separated by a 304-amino-acid open reading frame (unk) whose predicted sequence resembles that of the 2-hydroxy acid dehydrogenases . C . glutamicum mutants that carry null alleles of proB, proA, and proC were constructed or isolated from mutagenized cultures . Single proC mutants are auxotrophic for proline and secrete delta1-pyrroline-5-carboxylate, which are the expected phenotypes of bacterial proC mutants . However, the phenotypes or proB and proA mutants are unexpected . A proB mutant has a pleiotropic phenotype, being both proline auxotrophic and affected in cell morphology . Null proA alleles still grow slowly under proline starvation, which suggests that a proA-independent bypass of this metabolic step exists in C . glutamicum . Since proA mutants are complemented by a plasmid that contains the wild-type asd gene of C . glutamicum, the asd gene may play a role in this bypass.

Rev Inst Med Trop Sao Paulo, 1996 Jul-Aug, 38(4), 299 - 302
A single method to stain Malassezia furfur and Corynebacterium minutissimum in scales; Padilha-Goncalves A; The scales are collected by pressing small pieces of scotch tape (about 4 cm length and 2 cm width) onto the lesions and following withdrawal the furfuraceous scales will remain on the glue side . These pieces are then immersed for some minutes in lactophenol-cotton blue stain . Following absorption of the stain the scales are washed in current water to remove the excess of blue stain, dried with filter paper, dehydrated via passage in two bottles containing absolute alcohol and then placed in xylene in a centrifugation tube . The xylene dissolves the scotch tape glue and the scales fall free in the tube . After centrifugation and decantation the scales concentrated on the bottom of the tube are collected with a platinum-loop, placed in Canada balsam on a microscopy slide and closed with a cover slip . The preparations are then ready to be submitted to microscopic examination . Other stains may also be used instead of lactophenol-cotton blue . This method is simple, easily performed, and offers good conditions to study these fungi as well as being useful for the diagnosis of the diseases that they cause.

Biokhimiia, 1996 Jul, 61(7), 1294 - 302
{Participation of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate in reactions of bacteria on oxidative stress and their persistence in macrophages}; Ogrel' OD et al.; In aerated medium, Corynebacterium ammoniagenes cells accumulate 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEC) during heat shock and in the presence of O2--generating compounds or ozone . The ability to accumulate MEC was genetically transformed from C . ammoniagenes to E . coli XL-1; transformed E . coli (2-31 clone) accumulates MEC in the presence of glucose and glucose oxidase (generation of H2O2) or benzylviologen (generation of O2-); the viability of transformed bacteria inside the murine peritoneal macrophages also significantly increases . However, model conditions of phagosomes of warm-blooded animals (NO + H2O2 + O2-) did not cause MEC accumulation by C . ammoniagenes but increased the formation of polyphosphate which can be due to selective oxidative aberration of biosynthetic processes . Growth rate of Acanthamoeba castellanii on solid medium with bacterial lawn was not significantly different in C . ammoniagenes, C . ammoniagenes with preaccumulated MEC, E . coli XL-1, and E . coli 2-31 and did not depend on the accumulation of MEC by bacteria . Unlike the recipient E . coli strain, the transformed 2-31 clone synthesizes two nonpolar lipids (Rf = = 0.85 and 0.75; TCL on Silufol in hexane) and carotinoid pigments; this can be due to changes in metabolic pathways of isopentenylpyrophosphate that can be a precursor of MEC biosynthesis . Thus, MEC is involved in bacterial responses to certain components of oxidative stress and in bacterial persistence inside the macrophages.

J Clin Microbiol, 1996 Jul, 34(7), 1711 - 6
Heterogeneity of diphtheria toxin gene, tox, and its regulatory element, dtxR, in Corynebacterium diphtheriae strains causing epidemic diphtheria in Russia and Ukraine; Nakao H et al.; Diphtheria toxin (tox) and its regulatory element (dtxR) from 72 Corynebacterium diphtheriae strains isolated in Russia and Ukraine before and during the current diphtheria epidemic were studied by PCR-single-strand conformation polymorphism analysis (PCR-SSCP) . Twelve sets of primers were constructed (eight for tox and four for dtxR), and three regions within tox and all four regions of dtxR showed significant variations in the number and/or sizes of the amplicons . Two to four different SSCP patterns were identified in each of the variable regions; subsequently, tox and dtxR could be classified into 6 and 12 different types, respectively . The great majority of epidemic strains from both Russia and Ukraine had tox types 3 and 4, and only in a single preepidemic strain isolated in Russia were all eight tox regions identical to those of C . diphtheriae Park-Williams No . 8 (tox type 1) . Epidemic strains from Ukraine can easily be identified by dtxR type 5, while the majority of the Russian epidemic strains have dtxR of types 2 and 8 . No differences in the tox regions between mitis and gravis biotype strains were observed . However, dtxR types 2, 5, and 8 were identified only in the gravis biotype, and dtxR type 1 was characteristic for the mitis biotype strains . PCR-SSCP is a simple and rapid method for the identification of variable tox and dtxR regions that allows for the clear association of tox and dtxR types with strains of distinct temporal and/or geographic origins.

FEMS Microbiol Lett, 1996 Jul 1, 140(2-3), 247 - 51
Improved electro-transformation of highly DNA-restrictive corynebacteria with DNA extracted from starved Escherichia coli; Ankri S et al.; Differences of up to 33 000-fold in electro-transformability of highly DNA restrictive corynebacteria are observed in the DNA of a shuttle plasmid extracted from Escherichia coli hosts propagated in different nutritional conditions . Growth of the host in minimal medium increases plasmid transformability, whereas growth on rich media decreases it . In the E . coli DH5 alpha host, the starvation-dependent increase DNA transformability is reverted by supplementing with methionine, an obligate 5-adenosyl-methionine (SAM) precursor . This suggests that an E . coli nutritionally modulated SAM-dependent DNA-methyltransferase may be involved in this phenomenon.

FEMS Microbiol Lett, 1996 Jul 1, 140(2-3), 221 - 5
Extended host range of a beta-related corynebacteriophage; Cianciotto NP et al.; Unlike most beta-related phages isolated from Corynebacterium diphtheriae, phage 782 readily plaqued on strains of C . ulcerans and C . pseudotuberculosis . The extended host range of phage 782 was not, however, due to a greater ability of the phage to absorb to bacterial surfaces . Using chemical mutagenesis, a number of phage mutants were isolated which had diminished capacities to infect C . ulcerans, suggesting the existence of a locus (ehr) for extended host range . Pre-lysogenization of C . ulcerans strains with phage 782, but not its mutant form or other beta-related phages, rendered them susceptible to infection by previously excluded phages . An examination of recombinant between phages 782, pie, and beta localized ehr to a 7 kilobase region of DNA including attP . The data are compatible with the notion that ehr encodes an anti-restriction function.

Biochim Biophys Acta, 1996 Jun 3, 1307(1), 13 - 6
Identification of a third thioredoxin gene from Corynebacterium nephridii; Lim CJ et al.; We identified and sequenced a gene encoding a third thioredoxin (C3) from Corynebacterium nephridii . The determined nucleotide sequence encodes a thioredoxin of 145 amino acid residues, which is larger than most thioredoxins found in microbial cells and contains 6 cysteine residues . C . nephridii thioredoxin C3 is able to serve as a subunit of T7 DNA polymerase . C . nephridii is the first nonphotosynthetic procaryotic organism known to carry three different thioredoxins.

Southeast Asian J Trop Med Public Health, 1996 Jun, 27(2), 274 - 8
Immunity to diphtheria in women of childbearing age in Delhi in 1994: evidence of continued Corynebacterium diphtheriae circulation; Singh J et al.; Blood samples from 171 full-term pregnant women (aged 18-38 years) of middle socioeconomic status from Delhi were tested for diphtheria antitoxins by indirect hemagglutination (IHA) test . History of primary immunization/clinical diphtheria during childhood was not ascertainable, but none had been revaccinated against diphtheria at any time . About 94% women had very high antitoxin titers (> or = 0.125 IU/ ml); none had antitoxin titer less than 0.015 IU/ml, the minimum protective level . The titers were uniformly high in all age groups . However, women having 2 or more children had significantly higher antitoxin titers than those having no or one child (p < 0.01) . The results from this study and historical data on diphtheria in Delhi are compatible with continued transmission of C . diphtheriae in recent times in Delhi which is of sufficient magnitude to boost the antitoxin levels in adults, especially mothers having two or more children . The study highlights the need of increasing the immunization coverage with DPT among children to reduce the transmission of Corynebacterium diphtheriae.

Free Radic Res, 1996 Jun, 24(6), 473 - 81
Detection of a stable free radical in the B2 subunit of the manganese ribonucleotide reductase (Mn-RRase) of Corynebacterium ammoniagenes; Griepenburg U et al.; Ribonucleotide reductases catalyze the irreversible reductive formation of 2'-deoxyribonucleotides required for DNA replication and cell proliferation, and a radical mechanism was assumed to be involved in this reaction . In order to search for a radical in the aerobic manganese ribonucleotide reductase (Mn-RRase) by electron paramagnetic resonance (EPR) the native metal-containing 100 kDa B2 subunit was deliberately prepared from the wild type strain Corynebacterium ammoniagenes ATCC 6872 . Enrichment by 2'5'-ADP Sepharose 4B affinity chromatography, fast protein liquid chromatography (FPLC) with SuperoseTM12 and concentration by vacuum evaporation allowed for the first time the detection of a stable free radical by EPR spectroscopy at 77 K . The EPR spectrum exhibits an easily saturable doublet of 1.8 mT splitting and a line width of 1.3 mT at g = 2.0040 . The EPR signal intensity showed a clear correlation with the enzymatic activity upon long-time storage at ambient temperature (294 K) and inactivation by the specific RRase inhibitor hydroxyurea (HU) . This leads to the assumption of a protein-linked radical, with functional significance, in the metal-containing 100 kDa B2 subunit of the MnRRase of Corynebacterium ammoniagenes.

Can J Microbiol, 1996 Jun, 42(6), 593 - 603
Characterization of bacterial communities in heavy metal contaminated soils; Roane TM et al.; Heavy metal pollution is a principle source of environmental contamination . We analyzed heavy metal impacted soil microbial communities and found that, in general, although lead adversely affected biomass, metabolic activity, and diversity, autochthonous lead- and cadmium-resistant isolates were found . In several metal-stressed soils, the microbial community consisted of two populations, either resistant or sensitive to lead . Additionally, a lead-resistant isolate was isolated from a control soil with no known previous exposure to lead, suggesting widespread lead resistance . Lead-resistant genera isolated included Pseudomonas, Bacillus, Corynebacterium, and Enterobacter species . Plasmids, ranging from 5 to 260 kb, were not detected through standard purifications from lead-resistant isolates . Positive correlations existed between antibiotic resistance and isolation habitat for lead-resistant strains, microbial metabolic activity and soil type, soluble lead concentration and microbial diversity, and arsenic concentration and total or viable cell concentrations.

Microbiologia, 1996 Jun, 12(2), 297 - 304
Genetic tools in pathogenic nocardioform actinomycetes; Navas J; Nocardioform actinomycetes are Gram-positive bacteria with high G+C content . Several species of Mycobacterium, Corynebacterium, Nocardia and Rhodococcus are important human or animal pathogens . Transposon mutagenesis and homologous recombination are powerful genetic tools used for the identification of pathogenicity determinants . Transposable elements with potential application to mutagenize pathogenic nocardioform actinomycetes are described . Homologous recombination experiments which have been recently achieved in mycobacteria and related actinomycetes are commented on.

Enferm Infecc Microbiol Clin, 1996 Jun-Jul, 14(6), 367 - 9
{Transient bacteremia caused by Corynebacterium urealyticum: apropos of 2 cases}; Buendia B et al.; BACKGROUND: Corynebacterium urealyticum is a pathogen mainly isolated from the urinary tract and seldom from the blood . We present two cases of bacteremia caused by multiresistant C . urealyticum isolated in two and three blood cultures respectively . PATIENTS AND METHODS: The two cases were studied . C . urealyticum was isolated from blood cultures and clinical charts were reviewed retrospectively . No history of prior antibiotic therapy was observed in either patient . Blood cultures were processed using BACTEC NC 730 system (Becton Dickinson) . The API Coryne system (BioMerieux) was used to identify both strains . RESULTS: Despite both patients having not received any antibiotic treatment, they improved clinically and microbiologically . Therefore, the episodes were considered as transitory bacteremias . CONCLUSION: Although C . urealyticum is not common, we believe that it is necessary to identify any diphtheromorphic microorganism in blood, when they are clinically significant.

Am J Pathol, 1996 Jun, 148(6), 1819 - 38
ICAM-1, VCAM-1, and MAdCAM-1 are expressed on choroid plexus epithelium but not endothelium and mediate binding of lymphocytes in vitro; Steffen BJ et al.; The expression of cell adhesion molecules (CAMs) in the choroid plexus was studied in normal brain and during experimental autoimmune encephalomyelitis (EAE) in the SJL/J mouse during inflammation induced by intracerebral injection of killed Corynebacterium parvum in the C3H/He mouse . Both ICAM-1 and VCAM-1, but not MAdCAM-1, were constitutively expressed on choroid plexus epithelium but not on the fenestrated capillary endothelial cells within the choroid plexus . During EAE, we observed an up-regulation of ICAM-1 and VCAM-1 and de novo expression of MAdCAM-1 on choroid plexus epithelial cells . In contrast, endothelial cells in the choroid plexus were not induced to express any of the investigated CAMs . In in situ hybridization analysis we demonstrated that ICAM-1, VCAM-1, and MAdCAM-1 were locally synthesized and that the amount of their mRNAs increased in the inflamed choroid plexus . In vitro, primary choroid plexus epithelial cells could be induced to express ICAM-1, VCAM-1, and MAdCAM-1 on their surface after treatment with proinflammatory cytokines such as tumor necrosis factor-alpha, interleukin-1, interferon-gamma, and lipopolysaccharide . To investigate the functional status of the expressed CAMs we performed Stamper-Woodruff binding assays on frozen sections of inflamed and naive brains . ICAM-1, VCAM-1, and MAdCAM-1 expressed in choroid plexus epithelial cells mediated binding of lymphocytes via their known ligands LFA-1 and alpha4-integrin, respectively . The expression of ICAM-1, VCAM-1, and MAdCAM-1 on choroid plexus epithelial cells together with the lack of their expression on the fenestrated choroid plexus endothelium raises the possibility that the epithelial blood-cerebrospinal-fluid barrier plays an important role in the immunosurveillance of the central nervous system.

Arch Microbiol, 1996 Jun, 165(6), 387 - 96
C3-carboxylation as an anaplerotic reaction in phosphoenolpyruvate carboxylase-deficient Corynebacterium glutamicum; Peters-Wendisch PG et al.; Phosphoenolpyruvate carboxylase (PEPCx) has recently been found to be dispensable as an anaplerotic enzyme for growth and lysine production of Corynebacterium glutamicum . To clarify the role of the glyoxylate cycle as a possible alternative anaplerotic sequence, defined PEPCx- and isocitrate-lyase (ICL)-negative double mutants of C . glutamicum wild-type and of the l-lysine-producing strain MH20-22B were constructed by disruption of the respective genes . Analysis of these mutants revealed that the growth on glucose and the lysine productivity were identical to that of the parental strains . These results show that PEPCx and the glyoxylate cycle are not essential for growth of C . glutamicum on glucose and for lysine production and prove the presence of another anaplerotic reaction in this organism . To study the anaplerotic pathways in C . glutamicum further, H13CO3--labeling experiments were performed with cells of the wild-type and a PEPCx-negative strain growing on glucose . Proton nuclear magnetic resonance analysis of threonine isolated from cell protein of both strains revealed the same labeling pattern: about 37% 13C enrichment in C-4 and 3.5% 13C enrichment in C-1 . Since the carbon backbone of threonine corresponds to that of oxaloacetate, the label in C-4 of threonine positively identifies the anaplerotic pathway as a C3-carboxylation reaction that also takes place in the absence of PEPCx.

Ann N Y Acad Sci, 1996 May 15, 782, 25 - 39
Construction of L-lysine-, L-threonine-, and L-isoleucine-overproducing strains of Corynebacterium glutamicum; Sahm H et al.; The gram-negative bacterium Corynebacterium glutamicum is used for the industrial production of amino acids, for example, of L-glutamate and L-lysine . By cloning and expressing the various genes of the L-lysine pathway in C . glutamicum, we would demonstrate that an increase of the flux of L-aspartate semialdehyde to L-lysine could be obtained in strains with increased dihydrodipicolinate synthase activity . Recently we detected that in C . glutamicum two pathways exist for synthesis of D,L-diaminopimelate and L-lysine . Mutants defective in one pathway are still able to synthesize enough L-lysine for growth, but the L-lysine secretion is reduced to 50 to 70% . Using NMR spectroscopy, we could calculate how much of the L-lysine secreted into the medium is synthesized via either one or the other pathway . Amplification of the feedback inhibition insensitive homoserine dehydrogenase and homoserine kinase in a high L-lysine-overproducing strain enabled channeling of the carbon flow from the intermediate aspartate semialdehyde towards homoserine, resulting in a high accumulation of L-threonine . For a further flux from L-threonine to L-isoleucine, the allosteric control of threonine dehydratase was eliminated.

Trop Anim Health Prod, 1996 May, 28(2), 158 - 62
Corynebacterium pseudotuberculosis infection and lymphadenitis (taloa or mala) in the camel; Afzal M et al.; Pure cultures of Corynebacterium pseudotuberculosis were obtained from 11 cases of lymphadenitis (known locally as taloa or mala) in camels . Camel isolates produced typical taloa in camels experimentally inoculated subcutaneously at the base of the external ear with 10(10) colony forming units . A sheep strain of C . pseudotuberculosis inoculated into camels produced a local abscess at the site of inoculation but did not produce taloa . Re-infection of camels recovered from experimental inoculation did not produce taloa suggesting the possibility of the development of a vaccine against lymphadenitis in camels.

J Dairy Sci, 1996 May, 79(5), 838 - 45
Blood selenium, vitamin E, vitamin A, and beta-carotene concentrations and udder health, fertility treatments, and fertility; Jukola E et al.; We investigated the activity of glutathione peroxidase in whole blood; concentrations of vitamin E, vitamin A, and beta-carotene in serum; SCC; udder bacterial infections and the incidence of clinical mastitis; fertility treatments; and the success of first AI of 511 dairy cows for 1 yr . The mean Se content in whole blood and the concentrations of vitamin E, vitamin A, and beta-carotene concentrations in serum were 191 micrograms/L, 5.9 mg/L, 0.39 mg/L, and 12.9 mg/L, respectively . An increase in Se concentration in whole blood was associated with a decrease in all infections, including infections by Staphylococcus aureus, Actinomyces pyogenes, and Corynebacterium spp . (-17.7, -31.7, and -70.6%, respectively) . There was no association among the different infections or SCC and concentrations of vitamin E, vitamin A, or beta-carotene, but an association existed between vitamin A concentration and SCC . The lower Se concentration in whole blood did not increase incidence of clinical mastitis . The Se concentration in whole blood (200 micrograms/L) was accepted as a target value to optimize udder health . The incidence of fertility disorders (anestrus, subestrus, cystic ovaries, or delayed ovulation) was 34.4% . The pregnancy rate following first insemination was 48.6% . No significant association was observed among Se in whole blood; concentrations of total vitamin E, vitamin A, or beta-carotene in serum; and fertility disorders or success of first AI.

Int J Urol, 1996 May, 3(3), 202 - 6
Detection of Proteus mirabilis urease gene in urinary calculi by polymerase chain reaction; Takeuchi H et al.; BACKGROUND: Urea-splitting microorganisms cannot always be detected by stone or urine culture in patients with infection stones . Detection of genetic elements within the calculi by the polymerase chain reaction (PCR) may be a useful alternative . In this study, we assessed the usefulness of the PCR method in detecting the urease gene specific to Proteus mirabilis in urinary calculi . METHODS: Thirty-eight metabolic stones (calcium oxalate and/or calcium phosphate, uric acid, or cystine) and 49 struvite stones were examined . The PCR was applied with DNA extracted by boiling pulverized stone pieces . RESULTS: Of the 87 stones, PCR demonstrated the presence of the P . mirabilis urease elements ureC1 and ureC2 in 17, all of which were struvite . Stone culture and urine culture had been performed in 22 and 46 struvite stone cases, respectively, and the PCR was positive in all of the 10 culture-positive calculi and also in two calculi from which P . mirabilis was not isolated . CONCLUSION: PCR was reliable and convenient for detecting P . mirabilis in desiccated struvite calculi . Study to detect other species such as Ureaplasma or Corynebacterium would be useful in elucidating the role of bacterial infection in the formation of these stones.

J Hosp Infect, 1996 May, 33(1), 71 - 6
Bacterial contamination of autologous bone marrow during processing; Smith D et al.; As part of an audit of the processing of autologous bone marrow, we found that marrow was often contaminated with organisms potentially pathogenic to neutropenic recipients . One of 14 marrows studied was found to be contaminated before the processing stage and five others became contaminated during processing . The organisms isolated at these stages were Propionibacterium sp., coagulase-negative staphylococci, Staphylococcus aureus and coryneforms, suggesting that the skin was the likely source of contamination . Five out of the 11 marrows returned to patients were found to be contaminated after thawing . Two of these were marrows previously shown to be contaminated with coagulase-negative staphylococci before freezing, and from these coagulase-negative staphylococci were isolated again, in one case the strains were indistinguishable . New organisms isolated after thawing included Bacillus sp . and Corynebacterium sporogenes suggesting contamination from the environment . No infections attributable to these organisms were demonstrated in any of the patients studied.

J Antimicrob Chemother, 1996 May, 37(5), 1005 - 9
In-vitro activity of psychiatric drugs against Corynebacterium urealyticum (Corynebacterium group D2); Munoz-Bellido JL et al.; We tested the in-vitro activity of amoxycillin, amoxycillin/clavulanic acid, cefotaxime, gentamicin, trimethoprim-sulphamethoxazole, tetracycline, norfloxacin, ciprofloxacin, vancomycin, teicoplanin, clindamycin and five psychiatric drugs (chlorpromazine, sertraline, fluoxetine, paroxetine and risperidone) against 32 strains of Corynebacterium urealyticum . Resistance rates exceeded 90% for all antibiotics except glycopeptides, quinolones and tetracycline . Sertraline was the most active psychiatric drug . We tested the influence of sertraline on the activity of amoxycillin, amoxycillin/clavulanic acid, cefotaxime, gentamicin, trimethoprim-sulphamethoxazole, tetracycline and ciprofloxacin . We did not observe antagonism in any case . Sertraline enhanced the activity of ciprofloxacin and tetracycline against all strains (MIC decrease: 4-64-fold for ciprofloxacin, 2-32-fold for tetracycline).

J Clin Microbiol, 1996 May, 34(5), 1290 - 2
Endocarditis of native aortic and mitral valves due to Corynebacterium accolens: report of a case and application of phenotypic and genotypic techniques for identification; Claeys G et al.; Endocarditis of native aortic and mitral valves due to an organism identified as Corynebacterium accolens developed in a 73-year-old patient without predisposing factors . The organism was identified as C . accolens by biochemical identification, amplified rRNA gene restriction analysis, and DNA-DNA hybridization . This is the first case of C . accolens endocarditis reported, adding to the increasing number of Corynebacterium-related cases of endocarditis.

J Clin Microbiol, 1996 May, 34(5), 1275 - 6
Misidentification of toxigenic Corynebacterium diphtheriae as a Corynebacterium species with low virulence in a child with endocarditis; Pennie RA et al.; A 6-year-old boy presented to a university hospital in Malaysia with infective endocarditis complicating cyanotic congenital heart disease . Blood cultures showed a gram-positive, aerobic, coryneform-like bacillus identified by the hospital laboratory as Corynebacterium xerosis, but a reference laboratory identified the organism as a toxigenic strain of Corynebacterium diphtheriae . The two laboratories concurred on all biochemical test results except for sucrose fermentation.

J Clin Microbiol, 1996 May, 34(5), 1124 - 8
Most Corynebacterium xerosis strains identified in the routine clinical laboratory correspond to Corynebacterium amycolatum; Funke G et al.; A comprehensive study was performed on 25 bacterial clinical isolates originally identified as Corynebacterium xerosis . Three reference strains of C . xerosis were also included in the study . On the basis of a variety of phenotypic characteristics tested, all strains could be divided into two separate clusters: reference strains ATCC 373 (the type strain of C . xerosis) and ATCC 7711 showed yellow-pigmented, dry, rough colonies, fermented 5-keto-gluconate, exhibited strong leucine arylamidase and alpha-glucosidase activities, produced lactate as the major end product of glucose metabolism, were susceptible to most of the 19 antimicrobial agents tested, and showed an inhibition zone around disks containing the vibriocidal compound O/129 . In contrast, the remaining 26 strains including reference strain NCTC 7243 as well as all clinical isolates formed white-grayish, dry, slightly rough colonies, did not ferment 5-keto-gluconate, exhibited only weak leucine arylamidase and no alpha-glucosidase activity, produced large amounts of propionic acid as the end product of glucose metabolism, and were resistant to most antimicrobial agents tested, including O/129 . Chemotaxonomic (cellular fatty acids, mycolic acids, and G+C content) and molecular genetic (16S rRNA gene sequence) investigations revealed that the strains of the second cluster unambiguously belonged to the species C . amycolatum . Our data suggest that most strains reported in the literature as C . xerosis are probably misidentified and correspond to C . amycolatum.

Clin Infect Dis, 1996 May, 22(5), 851 - 2
A necrotic soft-tissue lesion due to Corynebacterium urealyticum in a neutropenic child; Saavedra J et al.; Corynebacterium urealyticum has been associated mainly with infections of the urinary tract . Other infections due to this organism are highly unusual . We report what we believe is the first case of necrotic infection of soft tissue due to C . urealyticum in a neutropenic child who was previously treated with chemotherapy . The infection was cured when the patient was treated with vancomycin and surgical debridement . The increase in the number of neutrophils may also have contributed to the patient's recovery.

Microbiology, 1996 May, 142 ( Pt 5), 1297 - 309
Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif; Patek M et al.; Relatively limited information about promoter structures in Corynebacterium glutamicum has been available until now . With the aim of isolating and characterizing such transcription initiation signals, random Sau3A fragments of C . glutamicum chromosomal DNA and of the corynebacterial phage phi GA1 were cloned into the promoter probe vector pEKplCm and selected for promoter activity by chloramphenicol resistance of transformed C . glutamicum cells . The nucleotide sequence of ten chromosomal and three phage fragments was determined and the transcriptional start (TS) sites were localized by primer extension analyses . Additionally, the promoters of five previously isolated C . glutamicum genes were cloned and mapped . All of the isolated promoters were also functional in the heterologous host Escherichia coli . A comparative analysis of the newly characterized promoter sequences together with published promoters from C . glutamicum revealed conserved sequences centred about 35 bp (ttGcca) and 10 bp (TA.aaT) upstream of the TS site . The position of these motifs and the motifs themselves are comparable to the -35 and -10 promoter consensus sequences of other Gram-positive and Gram-negative bacteria, indicating that they represent transcription initiation signals in C . glutamicum . However, the C . glutamicum consensus hexamer of the -35 region is much less conserved than in E . coli, Bacillus, Lactobacillus and Streptococcus.

J Bacteriol, 1996 May, 178(9), 2656 - 61
NADPH-dependent reductive ortho dehalogenation of 2,4-dichlorobenzoic acid in Corynebacterium sepedonicum KZ-4 and Coryneform bacterium strainNTB-1 via 2,4-dichlorobenzoyl coenzyme A; Romanov V et al.; Corynebacterium sepedonicum KZ-4, described earlier as a strain capable of growth on 2,4-dichlorobenzoate (G.M . Zaitsev and Y.N . Karasevich, Mikrobiologiya 54:356-369, 1985), is known to metabolize this substrate via 4-hydroxybenzoate and protocatechuate, and evidence consistent with an initial reductive dechlorination step to form 4-chlorobenzoate was found in another coryneform bacterium, strain NTB-1 (W.J.J . van den Tweel, J.B . Kok, and J.A.M . de Bont, Appl . Environ . Microbiol . 53:810-815, 1987) . 2-Chloro-4-fluorobenzoate was found to be converted stoichiometrically to 4-fluorobenzoate by resting cells of strain KZ-4, compatible with a reductive process . Experiments with cell extracts demonstrated that Mg - ATP and coenzyme A (CoA) were required to stimulate reductive dehalogenation, consistent with the intermediacy of 2-chloro-4-fluoro-benzoyl-CoA and 2,4-dichlorobenzoyl-CoA thioesters . 2,4-Dichlorobenzoyl-CoA was shown to be converted to 4-chlorobenzoyl-CoA in a novel NADPH-dependent reaction in extracts of both KZ-4 and NTB-1 . In addition to the ligase and reductive dehalogenase activities, hydrolytic 4-chlorobenzoyl-CoA dehalogenase and thioesterase activities, 4-hydroxybenzoate 3-monooxygenase, and protocatechuate 3,4-dioxygenase activities were demonstrated to be present in the soluble fraction of KZ-4 extracts following ultracentrifugation . We propose that the pathway for 2,4-dichlorobenzoate catabolism in strains KZ-4 and NTB-1 involves formation of 2,4-dichlorobenzoyl-CoA, NADPH-dependent ortho dehalogenation yielding 4-chlorobenzoyl-CoA, hydrolytic removal of chlorine from the para position to generate 4-hydroxybenzoyl-CoA, hydrolysis to form 4-hydroxybenzoate, oxidation to yield protocatechuate, and oxidative ring cleavage.

J Clin Invest, 1996 May 1, 97(9), 2038 - 44
Alpha-melanocyte-stimulating hormone reduces endotoxin-induced liver inflammation; Chiao H et al.; Alpha-Melanocyte-stimulating hormone (MSH) is a potent anti-inflammatory agent in many models of inflammation, suggesting that it inhibits a critical step common to different forms of inflammation . We showed previously that alpha-MSH inhibits nitric oxide (NO) production in cultured macro-phages . To determine how alpha-MSH acts in vivo, we induced acute hepatic inflammation by administering endotoxin (LPS) to mice pretreated with Corynebacterium parvum, alpha-MSH prevented liver inflammation even when given 30 min after LPS administration . To determine the mechanisms of action of alpha-MSH, we tested its influence on NO, infiltrating inflammatory cells, cytokines, and chemokines . Alpha-MSH inhibited systemic NO production, hepatic neutrophil infiltration, and increased hepatic mRNA abundance for TNF alpha, and the neutrophil and monocyte chemokines (KC/IL-8 and MCP-1) . We conclude that alpha-MSH prevents LPS-induced hepatic inflammation by inhibiting production of chemoattractant chemokines which then modulate infiltration of inflammatory cells . Thus, alpha-MSH has an effect very early in the inflammatory cascade.

Biochemistry, 1996 Apr 23, 35(16), 5292 - 9
Sarcosine oxidase contains a novel covalently bound FMN; Willie A et al.; Sarcosine oxidase from Corynebacterium sp . P-1 is a heterotetrameric protein containing three different enzymes: noncovalent FAD, noncovalent NAD+, and covalently bound flavin which is released as 8 alpha-(N3-histidyl)riboflavin upon complete hydrolysis of the protein . The following results show that the covalent flavin is not at the FAD level, as previously proposed, but it is rather as 8 alpha-(N3- histidyl)FMN coenzyme . First, no AMP is released when the protein moiety is treated with phosphodiesterase or subjected to mild acid hydrolysis . The enzyme contains a total of 5 mol of phosphate . Only one phosphate is covalently bound . The other four phosphates are noncovalent and attributed to noncovalently bound FAD and NAD+ . The 31P NMR spectrum of native enzyme exhibits resonances due to a single phosphate monoester an two pyrophosphates . Only a resonance due to phosphate monoester is observed after removal of the noncovalent cofactors and proteolytic digestion of the protein moiety . The 8 alpha-(N3-histidyl)FMN found in corynebacterial sarcosine oxidase represents a novel type of covalent flavin . Studies with sarcosine oxidases from Arthrobacter sp . and Pseudomonas sp . show that these heterotetrameric enzymes also contain covalently bound FMN plus noncovalently bound FAD and NAD+, similar to corynebacterial sarcosine oxidase . In contrast, two monomeric sarcosine oxidases (from Bacillus sp . and an unidentified microorganism) were found to contain only covalently bound FAD.

Biochemistry, 1996 Apr 9, 35(14), 4485 - 91
Mechanism-based inhibition of ribonucleoside diphosphate reductase from Corynebacterium nephridii by 2'-C-methyladenosine diphosphate; McFarlan SC et al.; The interaction of the adenosylcobalamin-dependent ribonucleoside diphosphate reductase of Corynebacterium nephridii with 2'-C-methyladenosine diphosphate (2'-C-methylADP) has been investigated in more detail {Ong, S . P., McFarlan, S . C., & Hogenkamp, H . P . C . (1993) Biochemistry 32, 11397-11404} . This nucleotide analog partitioned between normal reduction to 2'-deoxy-2'-C-methyladenosine diphosphate and decomposition to adenine, 2-methylene-3(2H)-4-methylfuranone, and presumably pyrophosphate . Reaction of the reduced enzyme with 2'-C-methylADP caused the development of a chromophore at 318 nm that is characteristic of the modification of the enzyme by the furanone {Harris, G., Ator, M., & Stubbe, J . (1984) Biochemistry 23, 5214-5225} . Incubation of {5'-3H2}-2'-C-methylADP with reduced reductase resulted in the covalent incorporation of the radiolabel into the protein and into aquocobalamin . A similar incubation of the enzyme, the labeled nucleotide analog, and dithiothreitol resulted in the formation of three radioactive hydrophilic compounds . Mass spectroscopic analysis of one of these compounds showed the presence of 2-methylene-3(2H)-4-methylfuranone . 2'-Deoxy-2'-C-methylADP is a very effective promoter of the tritium exchange reaction between {5'-3H2}adenosylcobalamin and the solvent, confirming that the exchange reaction is an integral part of the overall reduction . All these observations are consistent with the proposal that 2'-C-methylADP serves as a substrate and a mechanism-based inhibitor of the ribonucleotide reductase of C . nephridii, indicating that the enzyme is able to catalyze the conversion of the nucleotide analog to a 2'-deoxy-2'-C-methyl-3'-ketonucleotide that can collapse to the reactive 2-methylene-3(2H)-4-methylfuranone . Surprisingly, 2'-C-methylADP did not serve as either a substrate or an inhibitor of the ribonucleoside diphosphate reductase of Escherichia coli.

MMWR Morb Mortal Wkly Rep, 1996 Apr 5, 45(13), 271 - 3
Diphtheria outbreak--Saraburi Province, Thailand, 1994; Codon usage in the Mycobacterium tuberculosis complex; Department of Molecular Biology, Uppsala University, SwedenThe usage of alternative synonymous codons in Mycobacterium tuberculosis (and M . bovis) genes has been investigated . This species is a member of the high-G+C Gram-positive bacteria, with a genomic G+C content around 65 mol% . This G+C-richness is reflected in a strong bias towards C- and G-ending codons for every amino acid: overall, the G+C content at the third positions of codons is 83% . However, there is significant variation in codon usage patterns among genes, which appears to be associated with gene expression level . From the variation among genes, putative optimal codons were identified for 15 amino acids . The degree of bias towards optimal codons in an M . tuberculosis gene is correlated with that in homologues from Escherichia coli and Bacillus subtilis . The set of selectively favoured codons seems to be quite highly conserved between M . tuberculosis and another high-G+C Gram-positive bacterium, Corynebacterium glutamicum, even though the genome and overall codon usage of the latter are much less G+C-rich.

Curr Microbiol, 1996 Apr, 32(4), 225 - 8
The ability of a recombinant Escherichia coli strain to synthesize 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate correlates with its tolerance to in vitro induced oxidative stress and to the bactericidal action of murine peritoneal macrophages; Ogrel OD et al.; A number of bacteria are able to synthesize 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (BOSS) in response to oxidative stress . Here we show that the ability to synthesize BOSS can be genetically transferred from Corynebacterium ammoniagenes to Escherichia coli . A total DNA library from C . ammoniagenes ATCC 6872 established in the pBluescript SKII+vector backbone was transfected into E . coli XL-1 blue . Recombinant clone 2-31, which was resistant to redox-cycling agents, was selected . NMR studies showed that this clone was able to synthesize BOSS . We also studied the resistance of clone 2-31 to the bactericidal action of macrophages . Clone 2-31 cells had better survival within murine peritoneal macrophages than parental E . coli XL-1-blue cells . Since the ability to synthesize BOSS correlates with increased survival of bacteria within macrophages, we suggest that the pathogenicity of Corynebacteria could be mediated through the synthesis of BOSS.

J Tenn Med Assoc, 1996 Apr, 89(4), 115 - 6
Pulmonary abscess due to Corynebacterium striatum; Batson JH et al.; Non-diphtheria corynebacteria are normal commensals of the skin and mucous membranes of humans . Increasingly, however, these saprophytic organisms are being recognized as pathogens . Patients infected with these bacteria typically have an underlying immunosuppressive process and/or an indwelling venous catheter . Pleuropulmonary infection with Corynebacterium striatum is rare . We present a patient with diabetes mellitus who developed an intrapulmonary abscess due to C . striatum.

Antimicrob Agents Chemother, 1996 Apr, 40(4), 930 - 3
Comparison of antimicrobial susceptibilities of Corynebacterium species by broth microdilution and disk diffusion methods; Weiss K et al.; Corynebacterium species are increasingly being implicated in foreign-body infections and in immunocompromised-host infections . However, there are no specific recommendations on the method or the criteria to use in order to determine the in vitro activities of the antibiotics commonly used to treat Corynebacterium infections . The first aim of our study was to compare the susceptibilities of various species of Corynebacterium to vancomycin, erythromycin, and penicillin by using a broth microdilution method and a disk diffusion method . Second, the activity of penicillin against our isolates was assessed by using the interpretative criteria recommended by the National Committee for Clinical Laboratory Standards for the determination of the susceptibility of streptococci and Listeria monocytogenes to penicillin . Overall, 100% of the isolates were susceptible to vancomycin, while considerable variations in the activities of erythromycin and penicillin were noted for the different species tested, including the non-Corynebacterium jeikeium species . A good correlation in the susceptibilities of vancomycin and erythromycin between the disk diffusion and the microdilution methods was observed . However, a 5% rate of major or very major errors was detected with the Listeria criteria, while a high rate of minor errors (18%) was noted when the streptococcus criteria were used . Our findings indicate considerable variations in the activities of erythromycin and penicillin against the various species of Corynebacterium . Because of the absence of definite recommendations, important discrepancies were observed between the methods and the interpretations of the penicillin activity.

J Clin Microbiol, 1996 Apr, 34(4), 918 - 23
Detection of Mycobacterium tuberculosis by PCR amplification with pan-Mycobacterium primers and hybridization to an M . tuberculosis-specific probe; Tevere VJ et al.; Nucleic acid amplification techniques such as the PCR are very useful in the rapid diagnosis of infections by Mycobacterium tuberculosis . However, recent studies have shown that the accuracy of results can vary widely when tests are performed with nonstandardized reagents . We have developed a PCR assay for the detection of M . tuberculosis that is both rapid and accurate . The assay reagents are standardized and quality controlled . False-positive results due to carryover contamination are prevented by the incorporation of dUTP coupled with uracil-N-glycosylase restriction . This assay also employs pan-Mycobacterium amplification primers, allowing for flexibility in the mycobacterial species that can be identified from a single amplification reaction . The amplification is very sensitive; amplification products generated from as few as three bacteria can be detected by agarose gel electrophoresis . DNAs isolated from 33 of 34 mycobacterial species tested were amplified efficiently . Only DNA from Mycobacterium simiae did not amplify . The amplification is also very specific . Amplification products were generated only from the DNAs of bacteria in closely related genera such as Corynebacterium . The nonmycobacterial amplicons do not pose a problem, as they do not hybridize to mycobacterium-specific probes . Hybridization of amplicons to an M . tuberculosis-specific probe allows for the unambiguous identification of M . tuberculosis complex organisms . The clinical performance of this PCR assay was evaluated against that of culture in 662 respiratory specimens . Sensitivities of 100 and 73.1% were obtained from smear-positive and -negative respiratory specimens, respectively . The corresponding specificities were 100 and 99.8% . The high sensitivity and specificity, coupled with the potential for detecting a wide range of mycobacteria, make this assay a useful tool in the clinical management of mycobacterial infections.

Arch Biochem Biophys, 1996 Apr 1, 328(1), 51 - 6
Destruction of cholera toxin receptor on HeLa cell membrane using microbial endoglycoceramidase; Yamamoto K et al.; The sensitivity of HeLa cells to cholera toxin decreased by Corynebacterium sp . endoglycoceramidase treatment . This endo-enzyme destroyed the cholera toxin receptor, ganglioside G(M1), on the cell surface membrane by liberating intact oligosaccharide from it, which was confirmed by the decrease of intracellular cAMP accumulation and the results of the analysis of released oligosaccharide with a combination of pyridylamination method and HPLC . Fluorescence microscopy using the immunofluorescence method revealed that the amount of cholera toxin attached to the cells decreased in endoglycoceramidase-treated cells . The enzyme acted on cellular glycosphingolipids without addition of any activator protein which is required by other similar enzymes . Corynebacterium endoglycoceramidase is a useful tool to elucidate the function of glycosphingolipids on the cell surface in situ.

J Infect Dis, 1996 Apr, 173(4), 1014 - 8
Reactogenicity and immunogenicity of a protein-conjugated pneumococcal oligosaccharide vaccine in older adults; Powers DC et al.; Healthy adults > or = 50 years old were immunized with either pentavalent Corynebacterium diphtheriae C7 (beta197) cross-reactive material (CRM197) protein-conjugated pneumococcal vaccine (CV) containing 10 microgram each of capsular oligosaccharides from serotypes 6B, 14, 18C, 19F, and 23F or with licensed (23-valent, 25 microgram/serotype) pneumococcal polysaccharide vaccine (PV) . Adverse reactions, predominantly local in nature, occurred in 20 of 23 CV recipients versus 13 of 23 PV recipients (P<.05) . Compared with mean postvaccination antibody concentrations in PV recipients, those induced by CV were not significantly different for serotypes 6B, 14, 18C, and 23F and were lower for 19F (P<.05) . Six months later, reimmunization with PV of subjects who had initially received CV elicited a slight boost in antibody concentrations to levels that were not significantly higher than those achieved after the primary vaccination or than those in persons given a single dose of PV . Pneumococcal vaccines containing protein-conjugated oligosaccharides may offer no advantage over currently licensed preparations containing unconjugated polysaccharides for immunization of healthy older adults.

Nat Struct Biol, 1996 Apr, 3(4), 382 - 7
Identification of the primary metal ion-activation sites of the diphtheria tox repressor by X-ray crystallography and site-directed mutational analysis; Ding X et al.; The diphtheria tox repressor, DtxR, is a 226 amino acid transition metal ion-activated regulatory protein that controls the expression of diphtheria toxin in toxigenic Corynebacterium diphtheriae . The previously solved three-dimensional DtxR structures have identified two potential metal ion binding sites which may play a role in the activation of DNA binding by the repressor . We have used both X-ray crystallographic and site-directed mutational analysis of DtxR(C102D)-Ni2+ complexes and DtxR to identify the metal ion-binding site which results in the activation of the repressor . We demonstrate that DtxR contains both a primary and an ancillary metal ion binding site . The primary site functions directly in the activation of DNA binding . In contrast, the ancillary site contributes weakly, if at all, to activation.

J Biol Chem, 1996 Mar 8, 271(10), 5398 - 403
Functional and genetic characterization of the (methyl)ammonium uptake carrier of Corynebacterium glutamicum; Siewe RM et al.; Under nitrogen starvation conditions, Corynebacterium glutamicum was found to take up methylammonium at a rate of 20 +/- 5 nmol.min-1.(mg dry weight)-1 . The specific activity of this uptake was 10-fold lower when growing the cells under sufficient nitrogen supply, indicating a tight regulation on the expression level . The methylammonium uptake showed Michaelis-Menten kinetics with an Km of 44 +/- 7 microM and was completely inhibited by the addition of 10 microM ammonium . This finding and the fact that methylammonium was not metabolized by C . glutamicum strongly suggests that the uptake carrier actually represents an ammonium uptake system . Methylammonium uptake was strictly dependent on the membrane potential . From the pH optimum and the accumulation of methylammonium in equilibrium, it could be deduced that only one net charge is transported and, thus, that methylammonium is taken up in its protonated form via an uniport mechanism . The amt gene encoding the (methyl)ammonium uptake system was isolated and characterized . The predicted gene product of amt consists of 452 amino acids (Mr = 47,699) and shows 26-33% identity to ammonium transporter proteins from Saccharomyces cerevisiae and Arabidopsis thaliana . According to the hydrophobicity profile, it is an integral membrane protein containing 10 or 11 membrane-spanning segments.

Presse Med, 1996 Mar 2-9, 25(8), 327 - 9
{Diphtheria: apropos of an epidemic}; Bricaire F; Since Ramon developed the anti-diphtheria anatoxin in 1924 widespread vaccination has almost eliminated diphtheria, but since the acquired immunity is anatoxic and not anti-bacterial, carriage of the causal agent Corynebacterium diphteriae remains possible . In 1990, 1214 declared cases of diphtheria inaugurated an epidemic which spread through Russia, Ukraine and neighboring countries and even reached a few subjects in Europe and North America . Mortality in Russia was 10.15 per 100000 cases in 1993 . Higher rates were observed in children . The question is raised as to the level of protection in Western countries despite generalized vaccination programs . In France, a recent survey showed that only 49.3% of the 1004 subjects evaluated had complete protection and 20.4% had no protection at all . While 95% of young adults in the 15 to 24 age range were protected, the rate of protection was below one-third in subjects over 65 . These results emphasize the importance of anti-diphtheria vaccination programs and continued surveillance in adult populations . Current French legislation requires vaccination before the age of 18 months but without any requirement for re-vaccination in adults . The current situation clearly demonstrates that the risk of a diphtheria epidemic still exists, even in our Western countries . To completely protect the population, the present vaccination policy should include re-vaccinations of the adult population every 10 years, as for tetanus . Use of a preparation containing a reduced dose of vaccine, given with the tetanus and polio booster shots, is to be recommended . Re-vaccination with DT or the reduced dose dT would also be indicated after injury instead of tetanus alone . Surveillance and typing of C . diphteriae strains isolated from clinical cases should also be maintained.

Zhonghua Zhong Liu Za Zhi, 1996 Mar, 18(2), 123 - 6
{Thoracoscopy in malignant pleural effusions}; Zhang D et al.; To assess the value of thoracoscopy in malignant pleural effusions, the procedure and results of thoracoscopy by using a fiberoptic bronchoscope and a rigid cold-light thoracoscope in 130 cases with malignant pleural effusion are reported . The overall diagnostic rate was 91.5% (119/130) . The malignant pleural mesothelioma in 24 cases and metastatic cancers in 95 cases were histopathologically confirmed . Talcum powder, tetracycline and Corynebacterium parvum were separately sprayed through thoracoscope into pleural cavity in 69, 10 and 10 patients, and the success rates of complete and lasting pleurodesis were 87.0%, 5/10 and 8/10 respectively . Postoperative complications included transient fever and chest pain, local subcutaneous emphysema in 6 cases and tumor seeding at thoracoscopy site in 4 cases . It is concluded that thoracoscopy is simple, safe, reliable and of high practical value in the diagnosis of malignant pleural effusions and in assessment before exploratory thoracotomy, and that transendoscopical administration of drugs for pleurodesis is a very effective method for controlling malignant pleural effusions . The efficacy of the talc poudrage is better than tetracycline and Corynebacterium parvum.

Vet Microbiol, 1996 Mar, 49(1-2), 1 - 9
Genetic differences between nitrate-negative and nitrate-positive C . pseudotuberculosis strains using restriction fragment length polymorphisms; Sutherland SS et al.; Corynebacterium pseudotuberculosis has been classified into two biotypes according to ability to breakdown nitrate (Biberstein et al., 1971) . Restriction enzyme analysis (REA) has shown to reflect this differentiation, but numerous bands generated by this technique make interpretation difficult (Songer et al., 1988) . Restriction fragment length polymorphism's (RFLP's) has become an accepted genetic tool and was used in this study to determine if differences in nitrate reduction and other phenotypic characteristics could be identified genetically . Thirteen C . pseudotuberculosis isolates from four species of domestic animals from different parts of the world were investigated for phenotypic and genetic differences . Three closely related bacteria, Corynebacterium ulcerans, Actinomyces pyogenes (previously C . pyogenes),and Rhodococcus equi (previously C . equi) were included in the study to determine if the RFLP bands were unique to C . pseudotuberculosis . All C . pseudotuberculosis isolates were positive for urease production . Some differences in maltose and sucrose fermentation ability and nitrate reduction were recorded . Genetic differences were identified between the nitrate-positive group and the nitrate-negative group using non-radioactive ribosomal RNA (rRNA) probes Southern blotted to restriction digests of ApaI, PstI, and SstI . A small number of bands were seen, with distinct differences between the nitrate-positive and the nitrate-negative strains . No genetic variations were seen between strains which reflected differences in carbohydrate fermentation . Strains isolated from different animal species and from different parts of the world could not be differentiated genetically using these three restriction enzymes.

Antimicrob Agents Chemother, 1996 Mar, 40(3), 720 - 6
In vitro antimicrobial activities and spectra of U-100592 and U-100766, two novel fluorinated oxazolidinones; Jones RN et al.; Two new fluorinated oxazolidinones, U-100592 and U-100766, were evaluated against more than 659 gram-positive and -negative organisms and compared with glycopeptides, erythromycin, clindamycin, clinafloxacin, and chloramphenicol . U-100592 and U-100766 were usually equally potent, but the MICs at which 90% of the isolates are inhibited (MIC90s) of U-100592 for some staphylococci and enterococci were slightly lower than those of U-100766 (1 versus 2 micrograms/ml) . The MIC90 of U-100592 and U-100766 for oxacillin-resistant Staphylococcus aureus was 2 micrograms/ml, the same as observed for oxacillin-susceptible strains . The oxazolidinone MICs for other Staphylococcus spp . were < or = 2 micrograms/ml (MIC50, 0.5 to 1 microgram/ml) . All enterococci were inhibited by < or = 4 and < or = 2 micrograms of U-100592 and U-100766 per ml, respectively . Against 152 vancomycin-resistant enterococci (five species), both compounds had a narrow range of MICs (0.25 to 2 micrograms/ml) and a MIC90 of 1 microgram/ml . Corynebacterium jeikeium, Bacillus spp., and all tested streptococci were inhibited (< or = 4 micrograms/ml) . Members of the family Enterobacteriaceae and other gram-negative bacilli were not susceptible (MIC50, > 64 micrograms/ml) to either oxazolidinone . Three potencies of U-100592 and U-100766 disks were tested (5, 15, and 30 micrograms), and acceptable correlations (r = 0.81 to 0.90) with the measured MICs were observed . Best discrimination of the tentatively susceptible organisms (MICs, < or = 4 micrograms/ml) was demonstrated with the 30-micrograms disk concentration . The oxazolidinones demonstrated a dominant bacteristatic action . These oxazolidinones (U-100592 and U-100766) appear promising for treatment of gram-positive organisms that demonstrate resistance to contemporary therapeutic agents.

Hum Gene Ther, 1996 Mar 1, 7(4), 525 - 9
Coexpression of interleukin-4 and B7.1 in murine tumor cells leads to improved tumor rejection and vaccine effect compared to single gene transfectants and a classical adjuvant; Cayeux S et al.; To improve the vaccine potency of gene-modified tumor cells, using retroviruses, we have expressed the B7.1 gene in J558L cells and a subline previously transfected with the gene for interleukin-4 (IL-4) . Complete longterm tumor eradication occurred in only 73-82% of syngeneic BALB/c mice injected with IL-4 or B7.1 transfectants or tumor cells mixed with the adjuvant Corynebacterium parvum . In contrast, none of the mice injected with J558-IL4/B7.1 cells developed a tumor, thus demonstrating that IL-4 and B7.1 together induced a more potent antitumor immune response compared to either molecule alone . Immunization/challenge experiments demonstrated that IL-4/B7.1 co-transfected cells possessed improved and tumor-specific vaccine potency when compared to single gene transfectants and, more importantly, to a tumor cell/C . parvum mixture . Furthermore, irradiation of vaccine cells almost completely abrogated the vaccine effect . Together, our results mean a step toward an improved tumor cell vaccine that acquires efficacy by the concerted action of IL-4 and B7.1 and the use of viable cells.

Am J Respir Crit Care Med, 1996 Mar, 153(3), 1047 - 55
Heat-killed Corynebacterium parvum enhances endotoxin lung injury with increased TNF production in guinea pigs; Tasaka S et al.; Corynebacterium parvum (CP) is known to increase susceptibility to endotoxin, which is associated with increased production of tumor necrosis factor (TNF) . We investigated the effect of CP-priming on the pathogenesis of acute lung injury caused by intratracheal Escherichia coli endotoxin (lipopolysaccharide {LPS}) . Guinea pigs were divided into four groups: (1) control (n=6), (2) CP-alone (n=6), (3) LPS-alone (n=6) and (4) CP + LPS (n=6) . A CP dose of 4 mg/kg was injected intraperitoneally 7 d before the study . Animals were observed for 4 h after intratracheal administration of 0.02 mg/kg of LPS . The lung wet-to-dry weight ratio (W/D), {125I} albumin concentration ratio of lung tissue to plasma (T/P) and of bronchoalveolar lavage (BAL) fluid to plasma (B/P) and differential cell count in BAL fluid were examined . In the LPS-alone group, neither excess lung water nor increased albumin leakage was observed . The CP + LPS group showed increased lung water and albumin leakage as compared with the other three groups (p<0.05) . We also observed increased cell counts in BAL fluid (p<0.05), in the CP + LPS group . The spleen weight was increased in guinea pigs pretreated with CP, indicating reticuloendothelial system (RES) activation . In the CP + LPS group, the TNF level was increased in both plasma and BAL fluid . We conclude that pretreatment with CP enhances LPS-induced acute lung injury in parallel with increasing TNF production, which suggests that the activation of mononuclear phagocytes contributes to increased susceptibility to intratracheal endotoxin in guinea pigs.

Biochem Biophys Res Commun, 1996 Feb 15, 219(2), 537 - 42
Cloning of m-fluorophenylalanine-resistant gene and mutational analysis of feedback-resistant prephenate dehydratase from Corynebacterium glutamicum; Chan MS et al.; Corynebacterium glutamicum was mutated by nitrosoguanidine and five m-fluorophenylalanine (mFP)-resistant mutants were isolated . The mutants were resistant to phenylalanine-mediated feedback inhibition of the prephenate dehydratase activity . Cloning and characterization of the mFP-resistant gene revealed that mutant prephenate dehydratase, encoded by the phe A gene, confers the mFP-resistant phenotype upon C . glutamicum . To determine the amino acid residues to which variation may result in the feedback resistance of prephenate dehydratase, the phe A gene was modified by site-directed mutagenesis and the activities of mutant enzymes were assayed in the presence of phenylalanine . The data indicated that Arg-202 and Gly-224 located at the C-terminal region of prephenate dehydratase were important residues regarding the feedback resistance . Variations of these residues rendered the enzyme insensitive to phenylalanine inhibition . The results also suggested that Gly-224 may reside at the entrance of phenylalanine-binding pocket.

J Endourol, 1996 Feb, 10(1), 31 - 4
Corynebacterium urealyticum (CDC Group D2) associated with staghorn calculus: treatment by percutaneous debulking and chemolysis; Nadler RB et al.; We report the formation of a staghorn calculus in a transplanted kidney caused by infection with a urea-splitting Corynebacterium group D2 organism . The stone was debulked percutaneously followed by intravenous vancomycin administration and urinary acidification with oral acetohydroxamic acid, leading to clearance of nearly all of the stone.

Antimicrob Agents Chemother, 1996 Feb, 40(2), 497 - 9
Influence of external factors in resistance of Corynebacterium urealyticum to antimicrobial agents; Garcia-Bravo M et al.; Corynebacterium urealyticum is usually resistant to multiple antibiotics . We analyzed whether previous hospitalization and/or the use of antibiotics was a factor associated with the appearance of resistance to different antibiotics in C . urealyticum . Our findings suggest that resistant strains of C . urealyticum are likely to be acquired directly from the hospital environment and that the use of antibiotics in the hospital setting could favor the appearance of multiresistant strains.

J Clin Microbiol, 1996 Feb, 34(2), 409 - 12
Validation of catheter semiquantitative culture technique for nonstaphylococcal organisms; Dooley DP et al.; The catheter semiquantitative culture roll tip technique has been validated as a discriminator between non-catheter-related bacteremias and catheter-related bacteremias (CRBs) caused by Staphylococcus species . However, this technique has not been specifically validated when used for the evaluation of catheters infected with organisms other than staphylococci . We reviewed catheters that had been submitted for semiquantitative roll tip culture as well as hospital records to determine clinical correlates of infection . Local infection and CRB were defined by standard criteria . Catheter-related sepsis (CRS) was defined as fever, leukocytosis, or hypotension which resolved with catheter removal, without another source of infection . For 195 catheters from 93 patients, gram-negative rods and enterococci were present on 36, fungi were on 25, Corynebacterium species were on 5, Bacillus species were on 3, Staphylococcus species were on 79, and 41 demonstrated no growth . Of 21 episodes of CRB or CRS due to nonstaphylococcal organisms, only 1 (questionable) episode was due to a catheter with < 15 CFU (P < 0.05) . Eleven of these 21 episodes of CRB or CRS were due to gram-negative rods and enterococci, of which only the questionable episode was due to a catheter with < 15 CFU . Nine of these 21 episodes of CRB or CRS were due to fungi, none of which were associated with a catheter with < 15 CFU . The data for Staphylococcus species recapitulated published data (none of 21 CRB or CRS episodes were associated with catheters with < 15 CFU) and validated this retrospective technique . The data presented in this study validate the use of the semiquantitative culture technique for the evaluation of catheter-related infections caused by organisms other than staphylococci.

Tierarztl Prax, 1996 Feb, 24(1), 17 - 21
{Nephrectomy for chronic, unilateral suppurative pyleonephritis in cattle}; Hirsbrunner G et al.; This report presents the case history of a five-year-old Eringer cow suffering from chronic hematuria . Results of clinical examination, ultrasonography of the kidneys, endoscopy of the bladder, and cytologic and bacteriologic analysis of the urine revealed unilateral pyelonephritis of the right kidney caused by Corynebacterium renale . Antimicrobial treatment with procaine penicillin was not successful . Therefore, nephrectomy of the affected kidney was performed through a right flanc approach in the standing animal . Follow-up examination ten months after surgery revealed normal general condition . Etiopathogenesis and diagnostic and therapeutic procedures of pyelonephritis in cattle are discussed.

J Dairy Sci, 1996 Feb, 79(2), 334 - 6
Effects of freezing on the viability of nine pathogens from quarters with subclinical mastitis; Murdough PA et al.; Milk samples from 45 quarters containing mastitis pathogens were collected from lactating cows to determine the viability of those pathogens after freezing . An initial bacteria count was conducted, and samples were divided into 2-ml portions and frozen . Weekly bacteria counts were conducted for 6 wk . Viability after freezing was determined on five isolates of nine bacterial species: Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus chromogenes, Staphylococcus xylosus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Corynebacterium bovis, and Escherichia coli . Bacteria counts were converted to logarithm base 10, and analysis of variance was conducted to determine alterations in viability over the 6-wk period . Freezing of quarter milk samples for 6 wk did not affect viability of any of these pathogens.

Biochemistry, 1996 Jan 30, 35(4), 1137 - 49
Crystal structure of diphtheria toxin bound to nicotinamide adenine dinucleotide; Bell CE et al.; Diphtheria toxin (DT), a 58 kDa protein secreted by lysogenic strains of Corynebacterium diphtheriae, causes the disease diphtheria in humans by gaining entry into the cytoplasm of cells and inhibiting protein synthesis . Specifically, the catalytic (C) domain of DT transfers the ADP-ribose group of NAD to elongation factor-2 (EF-2), rendering EF-2 inactive . In order to investigate how the C-domain of DT binds NAD and catalyzes the ADP-ribosylation of EF-2, the crystal structure of DT in complex with NAD has been determined to 2.3 A resolution . This is the first crystal structure of an ADP-ribosyltransferase (ADP-RT) enzyme in complex with NAD and suggests the features of the ADP-RT fold which are important for NAD binding . The conformation of NAD in the complex and the proximity of the Glu148 carboxylate group of the C-domain to the scissile, N-glycosidic bond of NAD suggest plausible modes of catalysis of the ADP-ribosylation reaction . Residues 39-46 of the active-site loop of the C-domain become disordered upon NAD binding, suggesting a potential role for this loop in the recognition of the ADP-ribose acceptor substrate, EF-2 . The negatively charged phosphates and two ribose hydroxyls of NAD are not in direct contact with any atoms of the C-domain . Instead, they form an exposed surface which appears to be presented for recognition by EF-2 . Structural alignments of the DT-NAD complex with the structures of other members of the ADP-RT family suggest how NAD may bind to these other enzymes.

Med J Aust, 1996 Jan 15, 164(2), 72 - 5
Non-toxigenic Corynebacterium diphtheriae biovar gravis: evidence for an invasive clone in a south-eastern Australian community; Hogg GG et al.; OBJECTIVE: To determine the prevalence and clonality of non-toxigenic Corynebacterium diphtheriae biovar gravis in a community with two cases of endocarditis caused by this organism . SETTING: A Koorie (Aboriginal) community in Gippsland, eastern Victoria, in 1994 . METHODS: Nose and throat swabs were collected from 359 community contacts of the cases and cultured for C . diphtheriae . Strains isolated from the contacts were compared by pulsed-field gel electrophoresis (after digestion with Sma1, Not1 and Sfi1) with those from the invasive cases in the same community, another invasive case in Victoria, a cluster of invasive cases in New South Wales (NSW) (1990-1991), and other stored strains isolated from skin ulcers and sore throats . RESULTS: Non-toxigenic strains of C . diphtheriae biovar gravis were isolated from throat swabs of five of the case contacts . Uniform DNA patterns were found for the two community cases, the other Victorian case, nine of ten isolates from NSW, and the five throat isolates from case contacts . CONCLUSION: An invasive clone of C . diphtheriae biovar gravis appears to have been responsible for the three Victorian cases of endocarditis . It was also present among case contacts and responsible for previous invasive cases in NSW . Prophylactic treatment should be considered for clearly defined contacts in all instances where C . diphtheriae is isolated from a normally sterile site, regardless of the toxigenic nature of the strain.

J Immunother Emphasis Tumor Immunol, 1996 Jan, 19(1), 21 - 32
Secretion of both IL-2 and IL-4 by tumor cells results in rejection and immunity; Strome SE et al.; The generation of a therapeutic immune response to malignancy is critically dependent on the inherent immunogenicity of the tumor . Our study demonstrates that secretion of both interleukin-2 (IL-2) and IL-4 by a seemingly nonimmunogenic tumor abrogates tumorigenicity, and mice that have rejected the genetically modified tumor are immune to challenges with the parental tumor . The induction of immunity by the IL-2/IL-4-secreting tumor was significantly better than that achieved with the admixture of tumor cells and the classic adjuvant, Corynebacterium parvum . To elicit a primary immune response, the majority of cells needed to secrete both cytokines . Ad-mixture of IL-2-secreting cells with IL-4-secreting cells did not result in tumor cell rejection . The IL-2/IL-4-secreting tumor cells were efficiently rejected in animals immunosuppressed by total body irradiation . Depletion of CD4+ or CD8+ T cells did not abrogate rejection of the tumor cells, but the animals depleted of CD4 cells failed to generate protective immunity . Our study demonstrates that secretion of the combination of IL-2 and IL-4 significantly enhances tumor immunogenicity . The requirement of cells secreting both cytokines suggests an intricate mechanism different from the mere presence of both cytokines at the tumor-inoculation site.

Scand J Infect Dis, 1996, 28(1), 37 - 40
Diphtheria outbreak in St . Petersburg: clinical characteristics of 1860 adult patients; Rakhmanova AG et al.; An epidemic of respiratory tract diphtheria began in Russia in 1989 . In 1994 more than 2,500 cases occurred in St . Petersburg alone . We describe clinical findings in the 1,860 adult patients treated in Botkin's Hospital . The study is based on a retrospective review of patient records . In 98% of the patients the diagnosis was confirmed by a positive throat culture growing a toxin producing strain of Corynebacterium diphtheriae . A catarrhal disease without membranes was present in 1,256 (67.5%) patients, 150 patients had membranes on tonsils only, 268 patients on tonsils, the uvula, soft palate and posterior pharynx and 35 patients on larynx or in the lower respiratory tract . 42 patients (2.3%) died . Among the deceased patients 26 were alcoholics, whereby the death rate for non-alcoholics was probably around 1% . 151 patients (8.1%) had a toxic form of the disease with swelling of the neck . This form of the disease carried a high mortality, 25.7% . In a subgroup of 1,045 patients the protective efficacy of vaccination could be evaluated . A 2.2-fold protection was found, but the study may underestimate the efficacy . We conclude, that if a wide diphtheria epidemic affects an industrialized country, it would probably not any more be the big killer that it was in Europe and in the United States in the 1950's and 1960's.

Scand J Infect Dis, 1996, 28(6), 635 - 6
"Corynebacterium aquaticum" wound infection after high-pressure water injection into the foot; Larsson P et al.; The organisms presently named "Corynebacterium aquaticum" have their natural habitat in water and are increasingly often isolated in clinical specimens, but are very seldom the proven cause of infection . A case of a 24-year-old man with a "C aquaticum" wound infection secondary to a high-pressure water injection injury in the foot is described . Cefadroxil and cefuroxime were used for treatment.

Schweiz Arch Tierheilkd, 1996, 138(12), 596 - 9
{Isolation of Corynebacterium diphtheriae subsp . belfanti from a cow with chronic active dermatitis}; Corboz L et al.; Diphtheria is an acute communicable disease of man caused by C . diphtheriae . Pharyngeal and cutaneous forms are described, where from both toxigenic and nontoxigenic strains can be isolated . The occurrence of C . diphtheriae in dairy cattle has already been reported in the past . The pathogens were isolated from ulcerated teats and from the milk of cows with mastitis as well . These animals were considered to play a role in the transmission of the disease to man . This paper describes the isolation and characterization of C . diphtheriae in a 4 years old cow with generalized, partly ulcerative and purulent skin lesions . Bacteriological examination revealed the presence of very numerous corynebacterium-like organisms, which were characterized as C . diphtheriae subsp . belfanti, a nontoxigenic subspecies of C . diphtheriae . The significance of C . diphtheriae in veterinary medicine and the possible role of cattle as a reservoir of these organisms are discussed.

Ann Pharm Fr, 1996, 54(5), 228 - 30
{Antiseptic properties of essential oil of Lippia sidoides Cham . Application to the cutaneous microflora}; Lacoste E et al.; Two samples of essential oils of Lippia sidoides Cham . have been tested for their antibacterial and antifungal properties against some microorganisms living on the skin of feet and armpits . The essential oils and also their main components, thymol and carvacrol, show strong antagonistic activities . Corynebacterium xerosis developing axillary odour is specially inhibited . But on the other hand no specific activities have been observed upon the feet microflora.

Acta Otolaryngol Suppl, 1996, 525, 51 - 5
Recent trends in clinical isolates from paranasal sinusitis; Suzuki K et al.; Trends in the detection of causative pathogens and changes in bacterial counts in patients with sinusitis treated between January 1989 and December 1993 were investigated . In adult patients with chronic sinusitis, Staphylococcus aureus (S . aureus), coagulase negative staphylococci (CNS), Streptococcus pneumoniae (S . pneumoniae), Corynebacterium sp., Haemophilus influenzae (H . influenzae), and Moraxella catarrhalis were often isolated while Pseudomonas aeruginosa (P . aeruginosa) and anaerobic bacteria were detected in 2.4% and 5.3% of patients, respectively . The bacteria isolated from adult patients with acute sinusitis and pediatric patients with either acute or chronic sinusitis were somewhat different from those of adult chronic sinusitis . No bacteria could be isolated from 5.8% of adult chronic sinusitis patients, 8.1% of adult acute sinusitis patients, and 3.1% of pediatric sinusitis patients . The detection rate for anaerobic bacteria has been rising in chronic sinusitis patients owing to improved detection techniques in recent years, while there has been no appreciable change in the isolation rate for other types of bacteria . When the pathogenicity of isolated bacteria was determined based on the amount of bacterial colonization it was found that P . aeruginosa, S . pneumoniae, H . influenzae, and S . aureus were significant as causative pathogens in sinusitis, while CNS.

J Gynecol Obstet Biol Reprod (Paris), 1996, 25(1), 27 - 32
{Granulomatous mastitis and corynebacteria infection . Two case reports}; Binelli C et al.; Diagnosis of granulomatous mastitis must be based on a multidisciplinary approach . First, it's necessary to eliminate carcinomatous mastitis . Usually, the diagnosis is unknown except for tuberculous and sarcoidosis granulomatous mastitis . On observations in two cases of Corynebacterium granulomatous mastitis, we discussed the diagnosis and therapeutic approach . When there is a clinical suspicion of granulomatous mastitis, surgical biopsy with immediate histological analysis and bacteriological culture of mammary tissue should be performed . This multidisiplinary approach should reduce the number of idiopathic granulomatous mastitis observed . Antibiotic treatment is required after biopsy or surgical excision of granuloma.

Microbiol Immunol, 1996, 40(1), 1 - 4
Different rifampicin inactivation mechanisms in Nocardia and related taxa; Tanaka Y et al.; Mycolic acid-containing bacteria inactivate rifampicin in a variety of ways such as glucosylation, ribosylation, phosphorylation and decolorization . These inactivations were found to be a species-specific phenomena in Nocardia and related taxa . Gordona, Tsukamurella and fast-growing Mycobacterium modified rifampicin by ribosylation of the 23-OH group of the antibiotic . Such ribosylation was not observed in Rhodococcus and Corynebacterium, but phosphorylation of the 21-OH group of rifampicin was observed in one strain of Rhodococcus . Nocardia modified the antibiotic by glucosylation (23-OH group) and phosphorylation, but ribosylation was not observed.

Lung, 1996, 174(4), 207 - 24
Respiratory infections: community-acquired pneumonia and newer microbes; Reynolds HY; Respiratory infections, especially community-acquired forms of pneumonia (CAP), are challenging for clinicians because (1) a causative microorganism can only be found in about 50% of cases; (2) initial therapy, therefore, must be based on a probable or most likely etiology in the context of the patient's overall medical condition; and (3) new microbes or those considered previously as normal flora or less virulent forms seem responsible for some cases . It is important to be acquainted with new causes of infection which include Legionella species, Chlamydia pneumoniae, diphtheroids in certain instances (Corynebacterium pseudodiphtheriticum), and viruses such as the Hanta strains . Infections with Bordetella pertussis are increasing . However, the ever present and most common cause of CAP, Streptococcus pneumoniae, continues to present problems because of increasing antibiotic resistance, the high case fatality rate when bacteremia accompanies pneumonia, and the inability to give prophylactic immunization to all people with risk factors for this infection.

Cancer Gene Ther, 1996 Jan-Feb, 3(1), 39 - 47
Therapeutic efficacy of T cells derived from lymph nodes draining a poorly immunogenic tumor transduced to secrete granulocyte-macrophage colony-stimulating factor; Arca MJ et al.; We examined the host immune response to the poorly immunogenic B16-BL6 melanoma, which was transduced to secrete granulocyte-macrophage colony-stimulating factor (GM-CSF) (450 ng/10(6)/24 h) . Tumor growth after subcutaneous inoculation was not significantly altered, although an influx of neutrophils and monocytes/macrophages was evident within tumors and draining lymph nodes (LNs) . Immunization with irradiated transduced cells did not induce systemic immunity to the parental tumor . However, vaccination with transduced tumors significantly augmented in vivo sensitization of draining LN cells . These tumor-draining LN (TDLN) cells, when secondarily stimulated in vitro with anti-CD3 monoclonal antibodies and expanded in interleukin-2 (10 U/ml), exhibited greater release of GM-CST and interferon-gamma against tumor compared with TDLN cells from animals with parental tumor . In adoptive immunotherapy, activated LN cells draining transduced tumors mediated significant reductions of the numbers of established pulmonary metastases compared with LN cells draining parental tumor, which were ineffective . In addition, the therapeutic efficacy of LN cells draining transduced tumors was significantly better than LN cells primed in vivo with tumor cells admixed with Corynebacterium parvum, which we have previously described as an approach to generate immune cells . Thus, GM-CSF appears to be an important adjuvant in the induction of tumor immunity.

Vet Res, 1996, 27(3), 295 - 303
Field trial evaluation of two teat dips containing nisin or polyvinylpyrrolidone iodophor designed for use before and after milking; Serieys F et al.; In a first trial involving six commercial dairy herds and 291 cows for a period of eight months, pre-milking udder sanitation by dipping teats in a 0.25% polyvinylpyrrolidone (PVP) iodophor product followed by wiping with paper towels was compared in each herd with traditional teat washing and wiping with individual udder cloths . The incidence of new intramammary infections by Staphylococcus aureus, Streptococcus uberis and Corynebacterium bovis were significantly (P < 0.05) reduced, respectively by 48%, 60% and 47% . There were no significant differences in the two groups of cows for other intramammary infections, total bacterial counts, clostridia spore counts, iodine residues in milk or teat condition scores . In a second trial involving nine commercial dairy herds and 367 cows for a period of seven months, a teat dip containing nisin used before and after milking was compared in each herd with a classical 0.5% iodophor product used in the same way . There was no significant difference in the incidence of new intramammary infections, in spite of a higher rate of new Staphylococcus aureus infections in the group of cows teat-dipped with the nisin product (P = 0.06) . It was concluded that pre-dipping with teat dips specifically designed to be safely used before milking can be more efficient than traditional pre-milking udder preparation . These teat dips, when used before and after milking, seem to be as efficient as products which should normally be restricted to post-milking use.

Medicina (B Aires), 1996, 56(1), 57 - 8
{Prosthetic valve endocarditis caused by Corynebacterium urealyticum}; Notario R et al.; Corynebacterium urealyticum has been recognised as causing inflammatory cystitis and other human infections . In our knowledge this is the first case of a prosthetic valve endocarditis due to C . urealyticum . It was diagnosed in a 61 year old male patient with a history of rheumatic fever, hypertension and aortic stenosis . He had undergone surgery to replace the aortic valve and to perform triple aortocoronary bypass . The isolate was not multiresistant . Endocarditis due to C . urealyticum is very rare . Corynebacterium species, usually considered as contaminants, frequently colonize surgical cardiovascular areas and must be taken into account as causative agents of severe endocarditis.

Indian J Exp Biol, 1996 Jan, 34(1), 48 - 52
Influence of fowl uropygial gland and its secretory lipid components on growth of skin surface bacteria of fowl; Bandyopadhyay A et al.; Bacterial species, which occur on the breast skin surface of adult (1 year old) white leghorn fowl with intact uropygial gland, were identified as : Staphylococcus epidermidis, Sarcina lutea, Streptomyces sp . and a facultative diphtheroid belonging to the genus Corynebacterium; S . epidermidis being the most predominant one . Two species of bacteria, namely, Staphylococcus aureus and Proteus sp . were shown to colonize the skin surface after 60 days of captivity . Extirpation of uropygial gland caused severe depletion of population of S . epidermidis, Streptomyces sp . and diphtheroid . The effect was more conspicuous after 60 days compared to that after 30 days of the gland removal . On the skin surface of glandless fowls the population of S . aureus increased significantly and a new form identified as anthracoid bacillus became the most predominant species after 60 days . Addition of total lipids from the free-flowing fowl uropygial secretion, as 0.2% suspension, to trypticase soya broth cultures of individual bacteria of fowl skin surface encouraged strongly the growth of S . epidermidis, Streptomyces sp . and Proteus sp . but suppressed the population of the anthracoid . When identical amount of diester wax or wax alcohol of the secretion was supplemented to the culture, more or less similar result was obtained . Wax alcohol also had a mild inhibitory effect on Streptomyces sp . Wax acids, added to the culture (0.2%) suppressed population of all the bacterial forms except Proteus sp., while the hydrocarbon fraction, which also contained some amount of squalene, produce an opposite effect.

Plasmid, 1996 Jan, 35(1), 62 - 6
Electrotransformation of highly DNA-restrictive corynebacteria with synthetic DNA; Ankri S et al.; Highly DNA-restrictive Corynebacteria can be transformed with DNA made in vitro by PCR amplification of a sequence that contains the replication origin of pBL1, a plasmid common to many Corynebacteria . In all strains examined, the transformation efficiencies of PCR-synthetized DNA equal or improve the performances of heterologous DNA extracted from wild-type and dam(-)-dcm-strains of Escherichia coli . The transformation efficiencies obtained with PCR-made DNA may be high enough to permit its general application to experiments of gene integration.

Wien Klin Wochenschr, 1996, 108(9), 255 - 61
{Current treatment strategy in malignant pleural effusion}; Turler A et al.; Malignant pleural effusions are a grave consequence of advanced cancer disease . The successful suppression of pleural fluid reaccumulation can make a major contribution to the management and palliative care of patients with disseminated cancer . Many treatment concepts have been reported in the literature . The recommended therapy in malignant pleural effusions consists of intrapleural instillation of a sclerotic agent to produce pleurodesis . Different substances have been used, including tetracyclines, cytostatic agents, fibrin, talc, Corynebacterium parvum, cytokines and others . We reviewed the most frequently used techniques of pleurodesis in order to define the most effective treatment concept . In 15 prospective randomized trials the success rates varied from 13% with bleomycin to 100% with talc or Corynebacterium parvum . Talc was superior to other agents in 6 of 6, Corynebacterium parvum in 3 of 4 and bleomycin or tetracycline only in 3 of 8 studies . Adverse effects were frequently observed with cytostatic agents, but were very rare in the case of talc or fibrin instillation . Comparing the recently published data pleurodesis with talc appears to be the most effective treatment strategy, followed by Corynebacterium parvum, bleomycin and tetracycline.

Microbiology, 1996 Jan, 142 ( Pt 1), 99 - 108
Genes and enzymes of the acetyl cycle of arginine biosynthesis in Corynebacterium glutamicum: enzyme evolution in the early steps of the arginine pathway; Sakanyan V et al.; A cluster of arginine biosynthetic genes of Corynebacterium glutamicum ATCC 13032, comprising argJ, argB and argD as well as part of argC and argF, has been cloned by heterologous complementation of an Escherichia coli argE mutant . The gene order has been established as argCJBDF by sequencing the entire 4.4 kb cloned DNA fragment . The C . glutamicum argB gene can be transcribed in E . coli cells from an internal promoter located in the coding part of the preceding argJ gene, whereas transcription of the argJ gene appears vector-dependent . Expression of the corynebacterial argB gene is repressed by arginine in the native host but not in recombinant E . coli cells . Feedback inhibition of the corresponding N-acetylglutamate kinase activity was observed both in cell extracts of C . glutamicum and in recombinant E . coli argB auxotrophic strains . Extracts of E . coli cells carrying cloned corynebacterial DNA display an ornithine acetyltransferase activity (encoded by argJ) which alleviates the acetylornithinase (encoded by argE) deficiency of the enterobacterial host . In contrast to Bacillus stearothermophilus ornithine acetyltransferase which also exhibits acetylglutamate synthase activity, C . glutamicum ornithine acetyltransferase appears monofunctional . ArgA and ArgB proteins from different sources share highly significant similarities . The evolutionary implications of these data are discussed.

Int J Syst Bacteriol, 1996 Jan, 46(1), 88 - 93
Agromyces mediolanus sp . nov., nom . rev., comb . nov., a species for "Corynebacterium mediolanum" Mamoli 1939 and for some aniline-assimilating bacteria which contain 2,4-diaminobutyric acid in the cell wall peptidoglycan; Suzuki K et al.; In the course of identifying aniline-assimilating bacteria, researchers found some gram-positive strains that contain 2,4-diaminobutyric acid in their cell wall peptidoglycans and menaquinone 12 as the predominant menaquinone . "Corynebacterium mediolanum" and "Flavobacterium dehydrogenans" also are known to contain 2,4-diaminobutyric acid in their cell walls, as well as menaquinone 12, but the taxonomic position of these organisms has not been established previously . We found that the aniline-assimilating strains, together with "C . mediolanum" and "F . dehydrogenans," belong to a single species of the genus Agromyces, as determined by phenotypic characteristics, DNA-DNA relatedness data, and 16S ribosomal DNA sequence similarity data . The name Agromyces mediolanus sp . nov., nom . rev., comb . nov., is proposed for these organisms . The type strain of A . mediolanus is strain JCM 3346 (= ATCC 14004 = NCIMB 7206).

J Med Microbiol, 1996 Jan, 44(1), 35 - 40
Phosphorylcholine-containing antigens in bacteria from the mouth and respiratory tract; Gillespie SH et al.; Phosphorylcholine (PC)-containing antigens were sought in 269 bacterial isolates from the mouth and respiratory tract by an enzyme immunoassay method . Only 41 (15%) isolates were PC-positive and of these 29 (70%) were strains of Haemophilus influenzae . Other species that produced positive results included two of five isolates of Gemella haemolysans, two of five isolates of Micrococcus spp., and a single strain each of Bacillus sp., Corynebacterium jeikeium, Lactococcus sp . and H . parainfluenzae . The presence of PC-containing antigens in H . influenzae may be an important source of cross-reaction in antigen detection techniques that detect the C-polysaccharide antigen of Streptococcus pneumoniae in respiratory specimens and would result in false positive results.

Gastroenterology, 1996 Jan, 110(1), 210 - 20
Cell-generated nitric oxide inactivates rat hepatocyte mitochondria in vitro but reacts with hemoglobin in vivo; Fisch C et al.; BACKGROUND & AIMS: Nitric oxide forms inactive iron-nitrosyl complexes within hepatic mitochondria in vitro . However, when formed in vivo, NO might react instead with hemoglobin . The aim of this study was to compare the effects of cell-derived NO on rat hepatocyte mitochondria in vitro and in vivo . METHODS: First, hepatocytes were cultured in vitro for 24 hours under a porous membrane supporting macrophages that were stimulated by endotoxin . Second, hepatic macrophage hyperplasia was induced in vivo by preadministration of killed Corynebacterium parvum; 7 days later, rats received endotoxin and were killed after 6 hours . Third, mitochondria were exposed to sodium nitroprusside in vitro, washed, mixed with blood, and recovered . RESULTS: Iron-nitrosyl complexes and hepatocyte mitochondrial dysfunction were observed in the in vitro model and prevented by an NO synthase inhibitor . In the in vivo model, however, despite a 130-fold increase in plasma nitrate levels and formation of hemoglobin-NO complexes in blood, no iron-nitrosyl complex was detected in hepatic mitochondria, and hepatic mitochondrial function was not impaired . In the third model, mitochondria lost preformed iron-nitrosyl complexes when exposed to blood . CONCLUSIONS: Although NO reacts with hepatocyte mitochondria in vitro, in vivo it reacts with sinusoidal hemoglobin without detectable impairment of hepatic mitochondrial function.

Biochemistry, 1995 Dec 26, 34(51), 16703 - 7
Discovery of a third coenzyme in sarcosine oxidase; Willie A et al.; Denaturation of recombinant sarcosine oxidase or the natural enzyme isolated from Corynebacterium sp . P-1 with guanidine hydrochloride releases noncovalently bound FAD and a second UV-absorbing component (peak 2) which comigrates with NAD+ during reversed-phase HPLC . Both FAD and peak 2 are also found in extracts prepared by incubating sarcosine oxidase at 37 degrees C for 30 min, a procedure which causes partial (approximately 50%) release of the enzyme's noncovalently bound FAD . Peak 2 in the 37 degrees C extract is heat labile and decomposes upon boiling for 5 min at pH 8.0 . A similar instability was observed with NAD+ . Reaction of the 37 degrees C extract from sarcosine oxidase with phosphodiesterase yields nicotinamide mononucleotide, AMP, and FMN, as expected for a mixture containing NAD+ and FAD . Peak 2 was converted to NADH upon reaction of the 37 degrees C extract with yeast alcohol dehydrogenase in the presence of ethanol . Guanidine hydrochloride extracts, prepared from recombinant or natural enzyme, contain 1 mol of NAD+/mol of FAD . Since sarcosine oxidase contains 1 mol of noncovalently bound FAD, the results show that the enzyme also contains 1 mol of NAD+ . The NAD+ is tightly bound and is not lost during enzyme purification . It is not susceptible toward hydrolysis by NADase, reduction by alcohol dehydrogenase, or nucleophilic attack by cyanide . Unlike the flavins in sarcosine oxidase, NAD+ is not reduced by sarcosine and is not in redox equilibrium with the flavins.

Vet Rec, 1995 Dec 16, 137(25), 633 - 5
Failure of exit-race teat spraying to control Corynebacterium bovis colonisation; Hillerton JE et al.; When an automated exit-race teat sprayer replaced a conventional teat dip cup for the application of a disinfectant containing 0.5 per cent iodine, there was an increase in the level of intramammary infection by Corynebacterium bovis at drying off from approximately 25 per cent of quarters to approximately 75 per cent of quarters . When the peak level of infection had been reached half of the clinical mastitis in the herd was caused by C bovis, and these were recurrent and chronic infections . There was some evidence that the increase in C bovis infection increased the bulk milk cell count . There were no changes in the rates of infection by major pathogens or by coagulase-negative staphylococci, another important secondary pathogen . The reintroduction of teat dipping rapidly reduced the rate of mastitis infection and the level of infection was reduced to approximately 20 per cent of quarters in about 12 months.

Am J Physiol, 1995 Dec, 269(6 Pt 1), G861 - 6
Sources of arginine for induced nitric oxide synthesis in the isolated perfused liver; Pastor CM et al.; Hepatocytes can be stimulated to express high levels of inducible nitric oxide synthase (iNOS), which utilizes arginine for nitric oxide (NO) synthesis . Hepatocytes also synthesize and catabolize arginine, an intermediate in the urea cycle, raising the possibility that the urea pathway may provide substrate for hepatic NO synthesis . To identify the sources of arginine for iNOS, we measured the release of NO-2 + NO-3 and urea in isolated rat livers perfused in a recirculation model with a Krebs-Henseleit-bicarbonate buffer containing either no added amino acid, arginine, or precursors for urea synthesis . To induce iNOS expression, rats were injected with killed Corynebacterium parvum (C . parvum) or with endotoxin . In livers from C . parvum- and endotoxin-treated rats, we found that 1) an intracellular source of arginine exists that provides substrate to iNOS; 2) additional exogenous arginine increase NO synthesis, demonstrating that endogenous arginine is insufficient for maximal NO synthesis; and 3) an increase in the rate of endogenous arginine synthesis within the urea cycle is inefficient in increasing NO synthesis, demonstrating the independence of the two pathways in the liver.

Arch Oral Biol, 1995 Dec, 40(12), 1119 - 24
Identification of a lipoarabinomannan-like lipoglycan in Corynebacterium matruchotii; Sutcliffe IC; The oral organism Corynebacterium matruchotii was investigated for the presence of lipoteichoic acid, as this common polyanionic macroamphiphilic component of Gram-positive bacteria has been implicated in phenomena related to calcium binding . Phenol-water extraction followed by a small-scale, hydrophobic-interaction chromatography step yielded carbohydrate-containing preparations that were distinguished from lipoteichoic acid by their low phosphorus content . Subsequently, large-scale phenol-water extracts from each of three strains of C . matruchotii were purified by hydrophobic-interaction chromatography and shown to contain a heterogeneous lipoglycan fraction . The major fatty acids present were the same as for the whole-cell fatty acid profiles but differed in their relative amounts . Qualitative analysis of the lipoglycan fractions revealed similarities of carbohydrate composition with a previously characterized lipoglycan fraction from C . diphtheriae and with the lipoarabinomannan/lipomannans found in the genus Mycobacterium . The carbohydrate composition and the low phosphorus content indicated that lipoteichoic acid was absent from C . matruchotii . The calcium-binding properties of C . matruchotii therefore cannot be attributed to lipoteichoic acid.

Lab Anim Sci, 1995 Dec, 45(6), 652 - 6
Protective effects of macrophage-derived interferon against encephalomyocarditis virus-induced diabetes mellitus in mice; Hirasawa K et al.; The involvement of macrophages in protection against diabetes mellitus in mice of BALB/c (susceptible) and C57BL (resistant) strains infected with the B (non-diabetogenic) or D (highly diabetogenic) variant of encephalomyocarditis (EMC) virus was examined . Pretreatment with the B variant of EMC virus (EMC-B), avirulent interferon (IFN) inducer, or Corynebacterium parvum inhibited diabetes in BALB/c mice infected with the D variant of EMC virus (EMC-D) . Treatment of C57BL mice with carrageenan to compromise macrophage function rendered C57BL mice susceptible to EMC-D-induced diabetes . In macrophage culture for BALB/c mice, EMC-B induced IFN at an earlier stage than did EMC-D . The C57BL mouse-derived macrophages produced more IFN than did BALB/c mouse-derived macrophages after stimulation with EMC-D . Moreover, C . parvum increased IFN production in macrophage cultures from BALB/c mice, whereas carrageenan inhibited that in macrophage cultures from C57BL mice . These results suggest that IFN derived from macrophages may have an important role in protecting mice against EMC virus infection.

Vaccine, 1995 Dec, 13(18), 1785 - 92
Caseous lymphadenitis vaccine development: site-specific inactivation of the Corynebacterium pseudotuberculosis phospholipase D gene; Tachedjian M et al.; Vaccines for ovine caseous lymphadenitis (CLA) are currently formulated using partially purified, formalin inactivated phospholipase D (PLD) derived from Corynebacterium pseudotuberculosis culture supernatants . Chemical treatment has been a common and effective way of inactivating bacterial toxins for use in toxoid vaccines . Genetic inactivation of toxin genes using site-specific mutagenesis has the potential to improve this process by providing a safer and more cost-effective product . In the present study amino acid substitutions at the putative catalytic site and metal binding domain of the PLD protein had a profound affect upon PLD activity and secretion from C . pseudotuberculosis . Two mutated PLD analogues that were secreted to a level of 40% compared to the wild-type and retained minimal activity showed promise for development as recombinant CLA vaccines . Further work will be required to establish their suitability for commercialization.

J S Afr Vet Assoc, 1995 Dec, 66(4), 222 - 9
Artificial transmission of Bolo disease in woolled sheep and attempted characterisation of the causative unclassified Corynebacterium sp; Joubert JP et al.; Bacterial isolates (n = 38) previously cultured from sheep with Bolo disease were compared bacteriologically with known Corynebacterium spp . and Actinomyces spp . The isolates did not conform to any previously described species but closely resembled C . pseudodiptheriticum and C . urealyticum . More comprehensive tests are needed to classify this Corynebacterium sp . Bacterial cultures of this unclassified Corynebacterium sp . were used artificially to induce Bolo disease in Dohne Merino sheep (n = 20) . Ten sheep were kept at Middelburg in the Cape Midlands (Northern Cape) under arid conditions and another 10 at Queenstown in the Eastern Cape in a more humid climate . Two suspensions containing 2.8 x 10(5) Corynebacterium sp . (inoculum A) and 2.8 x 10(9) Corynebacterium sp . (inoculum B) respectively were used to infect each sheep on 9 different sites on the skin . One sheep died during the course of the experiment . Corynebacterium sp . established itself on 81 out of 171 inoculation sites of the remaining sheep and caused typical lesions of Bolo disease, clinically and pathologically . Bolo disease lesions developed slowly over 175 days at Middelburg and 287 days at Queenstown . Weather conditions were unfavourable to the development of fleece-rot and mycotic dermatitis . No difference was seen in lesion development between rams and ewes or between sheep with 5 months' wool growth and those which were shorn before inoculation . More lesions developed with the higher concentration of inoculum B (49 sites positive) as compared to inoculum A (32 sites positive).

Eur Respir J, 1995 Dec, 8(12), 2174 - 7
Corynebacterium parvum versus tetracycline as pleural sclerosing agents in rabbits; Vargas FS et al.; Tetracycline has been one of the most commonly used agents for producing a pleurodesis . However, it is no longer available due to more stringent requirements on the manufacturing process . The objective of this project was to determine whether Corynebacterium parvum is an effective sclerosant in an experimental model in rabbits . The following medications were instilled intrapleurally in anaesthetized male rabbits: tetracycline 35 mg.kg-1 or C . parvum 4 or 8 mg, all diluted with bacteriostatic saline solution . Twenty eight days after the instillation, the animals were sacrificed and the pleural spaces assessed macroscopically for evidence of pleurodesis and microscopically for evidence of fibrosis and inflammation . The intrapleural injection of C . parvum was ineffective in creating pleural fibrosis . The mean degree of pleurodesis in the 10 rabbits who received tetracycline was 3.5 +/- 0.7 (scale 0-4) whilst in the 10 rabbits that received 4 mg C . parvum it was 0.0 +/- 0.0, and in the 10 rabbits that received 8 mg C . parvum it was 0.5 +/- 0.8 . Based on this study, we recommend that C . parvum should not be used as a pleural sclerosant in patients with normal pleura.

J Surg Res, 1995 Dec, 59(6), 636 - 43
Electron transport chain activity in normal and activated rat macrophages; Reichner JS et al.; The pivotal role played by the macrophage in specific and nonspecific immunity suggests that the physiological status of the macrophage may effect the overall regulation of the host defense system . Many studies have evaluated macrophages as effector cells by examining expression of surface markers, cytokine release, or tumor killing in the presence of challenge to host defenses . In this report, the physiological parameter of mitochondrial respiration in freshly isolated rat macrophages is shown to be regulated upon activation in vivo . Assay conditions for the reduction of MTT {3-(4,5-dimethylthiazol-2)-2,5-diphenyltetrazolium bromide} in rat macrophages were optimized and used to quantitate electron transport chain activity as a measure of mitochondrial respiration . Corynebacterium parvum administration significantly increased the activity of the mitochondrial electron transport chain in both peritoneal (120% increase, 0.18 +/- .01 vs 0.40 +/- .03, P < 0.01) and liver macrophages (143% increase, 0.12 +/- .02 vs 0.30 +/- .06, P < 0.01) as detected by augmented MTT reduction . It is demonstrated further that MTT reduction is distinct from the respiratory burst activity of macrophages and supports the mitochondrial localization of intracellular MTT reduction in this cell type . These results demonstrate that electron transport chain activity is a physiological indicator of macrophage activation.

Appl Environ Microbiol, 1995 Dec, 61(12), 4477 - 9
Mutations in the trpD gene of Corynebacterium glutamicum confer 5-methyltryptophan resistance by encoding a feedback-resistant anthranilate phosphoribosyltransferase; O'Gara JP et al.; The trpD gene from tryptophan-hyperproducing Corynebacterium glutamicum ATCC 21850 was isolated on the basis of its ability to confer resistance to 5-methyltryptophan on wild-type C . glutamicum AS019 . Comparative sequence analysis of the genes from the wild-type AS019 and ATCC 21850 trpD genes revealed two amino acid substitutions at the protein level . Further analysis demonstrated that the trpD gene product from ATCC 21850, anthranilate phosphoribosyltransferase, was more resistant to feedback inhibition by either tryptophan or 5-methyltryptophan than its wild-type counterpart . It is proposed that phosphoribosyltransferase insensitivity to tryptophan in ATCC 21850 contributes to an elevated level of tryptophan biosynthesis.

Gene, 1995 Dec 1, 166(1), 117 - 9
Cloning and sequence analysis of the Corynebacterium diphtheriae dtxR homologue from Streptomyces lividans and S . pilosus encoding a putative iron repressor protein; Gunter-Seeboth K et al.; The iron-regulated promoter involved in desferrioxamine B synthesis of Streptomyces pilosus contains a region homologous to the iron repressor (DtxR)-binding site of the diphtheria toxin gene promoter in Corynebacterium diphtheriae {Gunter et al., J . Bacteriol . 175 (1993) 3295-3302} . Here, we report the cloning and sequencing of the putative Streptomyces iron repressor gene, homologous to dtxR of C . diphtheriae . The N-terminal 139 amino acids of the deduced protein are 73% identical to DtxR.

J Bacteriol, 1995 Dec, 177(24), 7255 - 60
Multicopy suppression by asd gene and osmotic stress-dependent complementation by heterologous proA in proA mutants; Serebrijski I et al.; Auxotrophic proA mutants of Escherichia coli were complemented by two different classes of Corynebacterium glutamicum genes . One of these was the asd gene . The E . coli asd gene also complements the same proA alleles . Complementation of proA by the asd+ gene requires a high asd dosage and the proB and the proC gene products . The reciprocal complementation pattern (asd by the proA+ gene) was not observed . This complementation appears to be due to multicopy suppression by a proline biosynthetic gene whose product was expected to play a negligible role in this pathway . The other class of complementing clones carries the C . glutamicum proA gene . Complementation of E . coli proA mutants by the C . glutamicum proA+ gene was optimal at high osmolarity.

Gene, 1995 Nov 7, 165(1), 67 - 70
Genomic organization of the mycobacterial sigma gene cluster; Doukhan L et al.; We have previously described sigma A and sigma B and their structural genes, mysA and mysB, respectively, in Mycobacterium smegmatis . We have now sequenced the corresponding regions in the M . tuberculosis and M . leprae chromosomes, and have found the two homologous genes . The chromosomal linkage and the deduced amino acid (aa) sequences of the two genes show very high similarity in the three species of mycobacteria . We also report the finding of two other open reading frames (ORF) in these clusters . orfX, which has an unknown function, is located between mysA and mysB . The other ORF, located downstream from mysB, encodes a homolog of DtxR, the iron regulatory protein from Corynebacterium diphtheriae (Cd).

Plasmid, 1995 Nov, 34(3), 229 - 33
Partial characterization of small plasmids from Corynebacterium renale; Nath N et al.; A naphthalene-degrading strain of corynebacteria, Corynebacterium renale, harbors multiple small plasmids designated pCR1, pCR2, pCR3, and pCR4 with sizes of 1.4, 3.2, 4.4, and 5.7 kb, respectively . Plasmid pCR1 of 1.4 kb is the smallest plasmid reported in this group of bacteria and is present in high copy number . Attempts to clone whole pCR1 in Escherichia coli were unsuccessful but two of its fragments (750 and 650 bp) could be separately cloned in it . The 4.4-kb plasmid, pCR3, bears considerable restriction pattern similarity to a 4.4-kb plasmid belonging to the pBL1 group of cryptic plasmid of corynebacteria but has no sequence homology, suggesting that pCR3 represents a new member of the 4.4-kb group of corynebacterial plasmids.

Mol Microbiol, 1995 Nov, 18(3), 391 - 9
Molecular cloning, sequence analysis, and functional characterization of the lipopolysaccharide biosynthetic gene kdtA encoding 3-deoxy-alpha-D-manno-octulosonic acid transferase of Chlamydia pneumoniae strain TW-183; Lobau S et al.; The gene kdtA of Chlamydia pneumoniae strain TW-183, encoding the enzyme 3-deoxy-alpha-D-manno-octulosonic acid (Kdo) transferase of lipopolysaccharide biosynthesis, was cloned and sequenced . A single open reading frame of 1314 bp was identified, the deduced amino acid sequence of which revealed 69% similarity and 43% identity with KdtA of Chlamydia trachomatis and Chlamydia psittaci . The gene was expressed in the Gram-positive host Corynebacterium glutamicum and the primary gene product was characterized as a multifunctional glycosyltransferase . Cell-free extracts generated in vitro the genus-specific epitope of Chlamydia composed of the trisaccharide alphaKdo(2-8)alphaKdo(2-4)alphaKdo . The results show that a single polypeptide affords three different glycosidic bonds, which is in contradiction to the dogma of glycobiology: 'one enzyme - one glycosidic bond'.

Clin Infect Dis, 1995 Nov, 21(5), 1334 - 6
Sepsis due to coryneform group A-4 in an immunocompromised host; Bizette GA et al.; Corynebacteria are more commonly being recognized as significant human pathogens . We describe a case of Coryneform group A-4 sepsis secondary to infection of a Hickman catheter in an immunocompromised man; the organism was identified by biochemical analysis conducted at the Louisiana State Reference Laboratory.

J Clin Microbiol, 1995 Nov, 33(11), 3061 - 3
Application of PCR for detection of toxigenic Corynebacterium diphtheriae strains isolated during the Russian diphtheria epidemic, 1990 through 1994; Mikhailovich VM et al.; A total of 250 Corynebacterium diphtheriae isolates from clinical cases and carriers in Russia were assayed by PCR directed at the A subunit of the diphtheria toxin gene to distinguish toxigenic from nontoxigenic strains; 170 strains were positive as indicated by the presence of the 248-bp amplicon . The results of this PCR assay were in complete concordance with those of the standard immunoprecipitation assay (Elek), and the PCR assay is a useful tool for rapid identification in clinical laboratories.

J Clin Microbiol, 1995 Nov, 33(11), 3031 - 3
Assessment of Difco ESP 384 blood culture system by terminal subcultures: failure to detect Cryptococcus neoformans in clinical specimens; Tinghitella TJ et al.; Terminal subcultures were performed on 1,162 5-day negative blood culture sets which had been monitored by the Difco ESP 384, a continuous-monitor blood culture system . Of these, 16 (1.4%) had growth upon terminal subculture . The isolates not detected by the Difco ESP 384 were Cryptococcus neoformans (eight isolates), Candida albicans (one isolate), Staphylococcus aureus (two isolates) coagulase-negative staphylococcus (three isolates), Bacillus sp . (one isolate), and Corynebacterium sp . (one isolate) . Acridine orange staining was performed on 200 randomly selected negative blood culture sets from the study group . Of these, two sets were positive and grew out C . neoformans, as did the terminal subculture . A review of patient's medical records indicated that many of these false-negative isolates were clinically insignificant . The Difco ESP 384 failed to detect 1.4% of the isolates in this study, 50% of which were C . neoformans, indicating a deficiency in the detection mechanism of the system . Further studies demonstrated that while these isolates (C . neoformans) grew in the Difco media, the system did not detect this growth when the standard 5-day protocol was used.

Br J Dermatol, 1995 Nov, 133(5), 801 - 4
A papular eruption secondary to infection with Corynebacterium jeikeium, with histopathological features mimicking botryomycosis; Jucgla A et al.; Corynebacterium jeikeium has been increasingly recognized as a pathogen, particularly in immunocompromised patients and in those with a prosthetic heart valve . Although cutaneous manifestations of C . jeikeium infection have been described, we have only found two case reports that give an histological description of the lesions . We present three patients with haematological malignancies who developed infection with C . jeikeium and a papular eruption . Skin biopsy disclosed similar histological features in all three patients, namely numerous Gram-positive bacteria enclosed in an eosinophilic matrix, with a minimal inflammatory response . C . jeikeium was cultured from cutaneous lesions in two cases.

J Immunol, 1995 Nov 1, 155(9), 4391 - 6
Macrophage activation by culture in an anoxic environment; Albina JE et al.; The extracellular amino acid composition of experimental wounds in rats during peak macrophage infiltration bears the imprint of the elevated arginase activity present in wound fluid: L-arginine is found in this space in concentrations markedly lower, and L-ornithine in concentrations markedly higher, than those that are detectable in plasma . No evidence, in the form of L-citrulline or NO2- accumulation, can be found at this time for nitric oxide synthase (NOS) activity . Wound-derived macrophages, however, metabolize L-arginine through both arginase and NOS in culture . Given the requirements of NOS for O2 and the reduced O2 tension in wounds, experiments were performed to determine the role of O2 availability on the metabolism of L-arginine by wound-derived macrophages . Results demonstrated that, beyond inhibiting NOS, culture of wound-derived macrophages in an anoxic environment provided an activation signal, markedly increasing total L-arginine metabolism, arginase activity, NOS protein content, and the release of TNF-alpha and IL-6 . Neither resident nor Corynebacterium parvum-elicited peritoneal macrophages responded to anoxic culture with increases in L-arginine utilization, arginase activity or, in the case of resident macrophages, in NOS protein content . The enhanced TNF-alpha and IL-6 release induced by anoxia in wound-derived macrophages was also found in resident peritoneal macrophages . Anoxia appears to act, then, as an inducer of activation-associated traits in macrophages obtained from different sites.

Infect Immun, 1995 Nov, 63(11), 4501 - 5
Changes to the ocular biota with time in extended- and daily-wear disposable contact lens use; Stapleton F et al.; Gram-negative bacteria may play a role in the etiology of certain soft contact lens (SCL)-related diseases . Contact lens (CL) wear may modify the normal ocular biota, providing a more favorable environment for potential pathogens . This study reports temporal changes in ocular biota in daily-wear (DW) and extended-wear (EW) disposable SCL use in experienced and neophyte wearers . Lid margin and bulbar conjunctival biota were sampled prior to CL fitting in 26 previous DW SCL users, 18 previous EW SCL users, and 26 neophytes . Wearers were fitted with an etafilcon A CL in one eye and a polymacon CL in the fellow eye . Lenses were worn on a daily basis by the 26 previous DW SCL wearers and on an EW basis by the remaining 44 subjects . The ocular biota was further sampled after 1, 3, 6, 9, and 12 months of wear . The ocular biota consisted of coagulase-negative staphylococci, Corynebacterium spp., Micrococcus spp., and Propionibacterium spp . Potential pathogens were rarely isolated at baseline . No significant trend of increasing ocular colonization was shown for extended CL wear . Lid and conjunctival colonization increased with DW SCL use (P < 0.001), although this increase occurred for nonpathogenic species only . Fewer potential pathogens were isolated from DW SCL than from EW SCL users (P < 0.05) . The lid margin consistently showed greater colonization than the conjunctiva and may be a source of potential pathogens during CL wear . Hydrogel CL wear appears to modify the ocular biota . An increased number of commensal organisms were present in DW SCL use . EW SCL use altered the spectrum of organisms isolated . These alterations may suppress the normal ocular defense mechanisms and may be relevant in the pathogenesis of CL-related disease.

Infect Immun, 1995 Nov, 63(11), 4284 - 9
Characterization of an iron-dependent regulatory protein (IdeR) of Mycobacterium tuberculosis as a functional homolog of the diphtheria toxin repressor (DtxR) from Corynebacterium diphtheriae; Schmitt MP et al.; The DtxR protein from Corynebacterium diphtheriae is an iron-dependent repressor that regulates transcription from the tox, IRP1, and IRP2 promoters . A gene from virulent Mycobacterium tuberculosis H37Rv was recently shown to encode a protein, here designated iron-dependent regulator (IdeR), that is almost 60% homologous to DtxR from C . diphtheriae . A 750-bp PCR-derived DNA fragment carrying the M . tuberculosis ideR allele was subcloned to both high- and low-copy-number vectors . In Escherichia coli, transcription from the C . diphtheriae tox, IRP1, and IRP2 promoters was strongly repressed by ideR under high-iron conditions, and ideR restored normal iron-dependent expression of the corynebacterial siderophore in the C . diphtheriae dtxR mutant C7(beta)hm723 . The M . tuberculosis IdeR protein was overexpressed in E . coli and purified to near homogeneity by nickel affinity chromatography . Gel mobility shift experiments revealed that IdeR bound to a DNA fragment that carried the C . diphtheriae tox promoter/operator sequence . DNAse I footprint analysis demonstrated that IdeR, in the presence of Cd2+, Co2+, Fe2+, Mn2+, Ni2+, or Zn2+, protected an approximately 30-bp region on DNA fragments carrying the tox, IRP1, or IRP2 promoter/operator sequences . IdeR reacted very weakly in Western blots (immunoblots) with antiserum against the C . diphtheriae DtxR protein, suggesting that the immunodominant epitopes of DtxR may be located in its poorly conserved carboxyl-terminal domain.

Infect Immun, 1995 Nov, 63(11), 4261 - 7
Protective vaccination with promastigote surface antigen 2 from Leishmania major is mediated by a TH1 type of immune response; Handman E et al.; Leishmania major promastigote surface antigen-2 complex (PSA-2) comprises a family of three similar but distinct polypeptides . The three PSA-2 polypeptides were purified from cultured promastigotes by a combination of detergent phase separation and monoclonal antibody affinity chromatography . Intraperitoneal vaccination of C3H/He mice with PSA-2 with Corynebacterium parvum as an adjuvant resulted in complete protection from lesion development after challenge infection with virulent L . major . Significant protection was also obtained in the genetically susceptible BALB/cH-2k and BALB/c mice . One of the PSA-2 genes was cloned and expressed in both Escherichia coli and Leishmania mexicana promastigotes . Vaccination with the recombinant PSA-2 purified from E . coli did not confer protection, in contrast to the L . mexicana-derived recombinant PSA-2, which provided excellent protection . CD4+ T cells isolated from the spleens of vaccinated mice produced large amounts of gamma interferon but no detectable interleukin 4 upon stimulation with PSA-2 in vitro . Limiting dilution analysis showed a marked increase in the precursor frequency of PSA-2-specific gamma interferon-secreting CD4+ T cells . No substantial change in precursor frequency was observed for interleukin 4-secreting T cells in the vaccinated mice . A CD4+ PSA-2 specific T-cell line generated from splenocytes of a vaccinated mouse produces a cytokine pattern consistent with a TH1 phenotype . Intravenous injection of this line into naive mice reduced significantly the parasite burden upon challenge infection . Taken together, the data suggest that vaccination with PSA-2 induces a TH1 type of immune response which protects mice from L . major infection . Moreover, a single recombinant PSA-2 polypeptide derived from a genomic clone can also vaccinate, provided that the structural form of the antigen is near native.

Am J Respir Cell Mol Biol, 1995 Nov, 13(5), 531 - 9
Deformability and CD11/CD18 expression of sequestered neutrophils in normal and inflamed lungs; Brown DM et al.; Neutrophil (PMN) sequestration in the pulmonary microvasculature precedes the migration of these cells into the airspaces in inflamed lungs . Intratracheal instillation of the heat-killed organism Corynebacterium parvum in the rat induces an alveolitis in which PMN constitute 70 to 80% of the total cell count in the bronchoalveolar lavage (BAL) . This acute alveolitis results in increased sequestration in the pulmonary microvasculature of 51Cr-labeled PMN when compared with control lungs . The aims of this study were to confirm this increased pulmonary PMN sequestration using unlabeled cells and to assess the function and adhesion molecule expression of such sequestered PMN . We counted the number of PMN and erythrocytes obtained by pulmonary vascular lavage (PVL) and compared the ratio of these two cell types in PVL and peripheral blood (PB) as a measure of the sequestration of PMN in the pulmonary vasculature . Compared with control animals, PVL in C . parvum-treated rats had higher PMN counts, which could not be accounted for by the PB leukocytosis . Sequestration of PMN in the pulmonary microvasculature depends on several factors, including the upregulation of adhesion molecules on both PMN and endothelial surfaces and the ability of the cells to deform when passing through the microcirculation . Cells obtained from the PVL were less deformable than PB cells in control but not in C . parvum-treated animals . The expression of the CD18 integrin on PMN obtained from the PVL of C . parvum-treated animals was increased compared with cells from control animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Thromb Res, 1995 Oct 15, 80(2), 113 - 23
Blood coagulation equilibrium in rat liver microcirculation as evaluated by endothelial cell thrombomodulin and macrophage tissue factor; Arai M et al.; The regulatory mechanisms of microcirculation might differ in the liver from other organs, because macrophages are resident in the hepatic sinusoids and sinusoidal endothelial cells are unique in shape and function . Thrombomodulin expression in endothelial cells and tissue factor activity in isolated macrophages were studied in the liver and lung of rats . In normal rats, the thrombomodulin expression was minimal in hepatic sinusoids, but prominent in pulmonary capillaries, while the tissue factor activity in the presence of endotoxin was higher in pulmonary macrophages than in Kupffer cells, although the levels in the absence of endotoxin were comparable in both cells . The tissue factor activity in hepatic macrophages was increased after priming of the cells with Corynebacterium parvum or after induction of liver necrosis or cirrhosis with carbon tetrachloride . In the necrotic or cirrhotic liver, increased thrombomodulin expression was seen along capillaries extending in necrotic areas and regenerating nodules, but this increase was minimal in the Corynebacterium parvum-treated rat liver . Blood coagulation equilibrium in microcirculation regulated by endothelial cells and macrophages may differ between the liver and lung . Such equilibrium in the liver may vary depending on pathological status.

Proc Natl Acad Sci U S A, 1995 Oct 10, 92(21), 9843 - 50
Structures of the apo- and the metal ion-activated forms of the diphtheria tox repressor from Corynebacterium diphtheriae; Schiering N et al.; The diphtheria tox repressor (DtxR) of Corynebacterium diphtheriae plays a critical role in the regulation of diphtheria toxin expression and the control of other iron-sensitive genes . The crystal structures of apo-DtxR and of the metal ion-activated form of the repressor have been solved and used to identify motifs involved in DNA and metal ion binding . Residues involved in binding of the activated repressor to the diphtheria tox operator, glutamine 43, arginine 47, and arginine 50, were located and confirmed by site-directed mutagenesis . Previous biochemical and genetic data can be explained in terms of these structures . Conformational differences between apo- and Ni-DtxR are discussed with regard to the mechanism of action of this repressor.

Pathologica, 1995 Oct, 87(5), 525 - 27
Empyema with malakoplakic-like lesions by Rhodococcus equi as a presentation of HIV infection; Calore EE et al.; Rhodococcus equi (Corynebacterium equi) is an aerobic actinomycetes, well described as a cause of pulmonary infection in different animals as horses, pigs and cows . This pathogen has a coccobacillar aspect and a variable acid-fast stain in tissues . Rare cases of human infection by Rhodococcus species were described, the majority by Rhodococcus equi, especially in patients with immunodeficiency syndrome (AIDS) in advanced stages of the disease . Usually the diagnosis of infections by Rhodococcus species is performed by positive blood or bronchoalveolar lavage cultures . Here we described a case of a pleuro-pulmonary infection by Rhodococcus equi, with malakoplakic-like lesions, that was the first manifestation of AIDS, whose diagnosis was performed by pleural biopsy (acid-fast bacteria with a variable coccobacillar aspect inside macrophages) and pleural fluid culture.

J Chemother, 1995 Oct, 7(5), 427 - 31
Etiology and therapy of chronic suppurative otitis; Campos MA et al.; Infectious diseases of the ear are important in adults due to their incidence and relapses . We carried out a study of aerobic microorganisms on 251 otic exudates from patients diagnosed as having chronic suppurative otitis media without cholesteatoma (119), chronic suppurative otitis media with cholesteatoma (85) and chronic external otitis (47) . The microorganisms predominantly isolated were, Pseudomonas aeruginosa, Staphylococcus aureus and other Enterobacteriaceae . 86% of isolates were monomicrobial and 14% of isolates were polymicrobial . In these latter the predominantly isolated microorganisms were also P . aeruginosa, S . aureus, Corynebacterium spp . and Proteus mirabilis . P . aeruginosa was the most commonly isolated and showed the highest percentages of resistance against antimicrobial agents tested . P . aeruginosa was most susceptible to ciprofloxacin and imipenem, but much less susceptible to cefotaxime, moxalactam and trimethoprim-sulfamethoxazole . S . aureus was highly sensitive to amoxicillin/clavulanate, trimethoprim-sulfamethoxazole, rifampin and teichoplanin . 100% of the isolates were resistant to penicillin G and ampicillin.

J Antimicrob Chemother, 1995 Oct, 36(4), 595 - 606
vanA genes in vancomycin-resistant clinical isolates of Oerskovia turbata and Arcanobacterium (Corynebacterium) haemolyticum; Power EG et al.; We report the cloning and sequencing of vanA genes present in the high-level vancomycin- and teicoplanin-resistant clinical isolates Oerskovia turbata 892 and Arcanobacterium (Corynebacterium) haemolyticum 872 . The presence of vanA was detected by Southern blotting and PCR and confirmed by DNA sequencing . vanA-like sequences were encoded on plasmids of 15 and 20 kb respectively . The A . haemolyticum 872 DNA sequence was identical to the published vanA sequence of vancomycin-resistant Enterococcus faecium BM4147, but the O . turbata 892 sequence showed three coding changes . Induction experiments indicated that vancomycin resistance in A . haemolyticum 872 and O . turbata 892 was constitutive . SDS-PAGE analysis of membrane proteins showed the presence of a c . 39 kD protein in both clinical isolates whose expression was unaltered in the presence of vancomycin, while a similar protein in E . faecium BM4147 was inducible . Since A . haemolyticum and O . turbata are naturally susceptible to vancomycin, the high-level constitutive resistance seen in these isolates appears to be mediated by vanA . This is the first report confirming the presence of vanA in genera other than Enterococcus.

Dtsch Tierarztl Wochenschr, 1995 Oct, 102(10), 408 - 9
Studies on the efficacy of combined immunostimulant-antibiotic therapy against experimental Mycoplasma gallisepticum infection in chickens; Hanafy MS et al.; Josamycin is an antibiotic known to become selectively concentrated intracellularly and in respiratory organs, the habitate of Mycoplasma gallisepticum . The aim of this present work was to evaluate the efficacy of josamycin when given alone or combined with an immunostimulant Cornebacterium cutis ultralysate . Groups of chickens were given josamycin alone or Corynebacterium ultralysate alone or both agents or nothing immediately before induction of Mycoplasma gallisepticum infection . Birds were subjected to pathological examination to evaluate the incidence and severity of air-sacculitis, bacteriological examination for re-isolation of Mycoplasma gallisepticum and for immunological examination to evaluate the humoral immune response to the infection (haemagglutination inhibiting titre determination) . The effect of treatments used in this study was to decrease the incidence and severity of air sacculitis . The magnitude of rise in haemagglutination inhibiting titres were greater and faster in birds given Corynebacterium ultralysate . Treatments failed to achieve complete elimination of Mycoplasma . No special advantage was obtained from the use of josamycin, its effects were rather similar to previously used chemotherapy.

Can J Microbiol, 1995 Oct, 41(10), 925 - 9
Comparison of 16S ribosomal RNA genes in Clavibacter michiganensis subspecies with other coryneform bacteria; Li X et al.; Nearly complete sequences (97-99%) of the 16S rRNA genes were determined for type strains of Clavibacter michiganensis subsp . michiganensis, Clavibacter michiganensis subsp . insidiosus, Clavibacter michiganensis subsp . sepedonicus, and Clavibacter michiganensis subsp . nebraskensis . The four subspecies had less than 1% dissimilarity in their 16S rRNA genes . Comparative studies indicated that the C . michiganensis subsp . shared relatively high homology with the 16S rRNA gene of Clavibacter xyli . Further comparison with representatives of other Gram-positive coryneform and related bacteria with high G+C% values showed that this group of bacteria was subdivided into three clusters . One cluster consisted of the Clavibacter michiganensis subsp., Clavibacter xyli, Arthrobacter globiformis, Arthrobacter simplex, and Frankia sp.; another cluster consisted of members of the corynebacteria-mycobacteria-nocardia (CMN) group of Mycobacteriaceae including Tsukamurella paurometabolum; and Propionibacterium freudenreichii alone formed a unique cluster, which was remote from other coryneform bacteria analyzed . The three clusters may reflect a systematic rank higher than the genus level among these bacteria.

Res Microbiol, 1995 Oct, 146(8), 633 - 41
Evaluation of the applicability of amplified rDNA-restriction analysis (ARDRA) to identification of species of the genus Corynebacterium; Vaneechoutte M et al.; The 16S rRNA genes (rDNA) of 50 strains belonging to 26 different coryneform bacterial species and genomospecies and of the type strain of Rhodococcus equi were enzymatically amplified . Amplified rDNA restriction analysis (ARDRA) with the enzymes AluI, CfoI and RsaI was carried out . The combination of the ARDRA patterns obtained after restriction with these three different enzymes enabled the differentiation between the following species: Corynebacterium accolens (number of strains = 2), C . afermentans subsp . afermentans (2), C . afermentans subsp . lipophilum (2), C . amycolatum (3), CDC coryneform group ANF-1-like (1), CDC coryneform group ANF-3-like (1), C . cystitidis (1), C . diphtheriae (4), C . jeikeium (3), C . macginleyi (2), C . minutissimum (1), C . pilosum (1), C . pseudotuberculosis (2), C . renale (2), C . striatum (2), C . urealyticum (3), C . xerosis (1), CDC coryneform groups B-1 (2), B-3 (2), F-1, genomospecies 1 and 2 (6), G, genomospecies 1 (1) and G, genomospecies 2 (2) . The following strains or species could not be differentiated from each other: C . pseudodiphtheriticum (2) from C . propinquum (former CDC coryneform group ANF-3) (2), CDC coryneform group F-1, genomospecies 1 (4) from genomospecies 2 (2) and C . jeikeium genomospecies A (1) from genomospecies C (2) . ARDRA may represent a possible alternative for identification of coryneforms, since this technique enabled the identification of most coryneforms tested and since DNA extraction (i.e . cell lysis by boiling), amplification, restriction and electrophoresis can be carried out within 8 hours . This might allow quick identification of C . diphtheriae and other possible pathogens of the genus Corynebacterium.

Can J Vet Res, 1995 Oct, 59(4), 306 - 10
Development of a specific biotinylated DNA probe for the detection of Renibacterium salmoninarum; Hariharan H et al.; A specific DNA probe for the identification of Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD), was developed from one of 3 clones pRS47, pRS49, and pRS26 of 5.1 kb, 5.3 kb, and 11.3 kb, respectively . The biotinylated pRS47/BamHI insert probe was tested on 3 dilutions of DNA extracted from 3 strains of R . salmoninarum and from 1 strain each of Arthrobacter protophormiae, Aeromonas salmonicida, Corynebacterium aquaticum, Carnobacterium piscicola, Listonella anguillarum, Micrococcus luteus, Pseudomonas fluorescens, Vibrio ordalii, and Yersinia ruckeri . In a dot blot assay, this probe hybridized only with the DNA from the R . salmoninarum strains . When used on kidney samples from fish challenged with R . salmoninarum, the dot blot hybridization assay with the probe was found to be as sensitive as culture . In a fluorescent antibody test, samples that were negative in culture and dot blot hybridization showed no more than one fluorescing cell in 50 microscopic fields examined . This DNA probe, therefore, has the potential for use in the diagnosis of BKD of fish.

J Clin Pathol, 1995 Oct, 48(10), 971 - 2
Corynebacterium aquaticum septicaemia in a neutropenic patient; Moore C et al.; Corynebacteria are a well recognised cause of sepsis in the immunocompromised patient . Corynebacterium aquaticum, however, is rarely seen in the clinical setting, being an environmental organism associated with fresh water . A septicaemic episode caused by this organism in a 74 year old neutropenic woman with an indwelling central venous catheter is reported . It is postulated that the source of the organism was untreated stored rainwater which she used for showering.

J Bacteriol, 1995 Oct, 177(20), 5991 - 3
Functional analysis of sequences adjacent to dapE of Corynebacterium glutamicum reveals the presence of aroP, which encodes the aromatic amino acid transporter; Wehrmann A et al.; An initially nonclonable DNA locus close to a gene of L-lysine biosynthesis in Corynebacterium glutamicum was analyzed in detail . Its stepwise cloning and its functional identification by monitoring the amino acid uptakes of defined mutants, together with mechanistic studies, identified the corresponding structure as aroP, the general aromatic amino acid uptake system.

J Appl Bacteriol, 1995 Oct, 79(4), 393 - 8
Antibacterial polyphenols from olive oil mill waste waters; Capasso R et al.; Olive oil vegetation waters (VW) were highly toxic to both phytopathogenic Pseudomonas syringae (Smith, Yung et al.) pv . savastanoi (Gram-negative) and Corynebacterium michiganense (Gram-positive) and showed bactericidal activity in their original concentration (in raw form) . Among the main polyphenols, present in the waste waters, methylcatechol proved to be the most toxic to Ps . savastanoi at 10(-4) mol l-1, and also demonstrated bactericidal activity, while on Coryne . michiganense it was only slightly active; catechol and hydroxytyrosol were less active on Ps . savastanoi, but inactive on Coryne . michiganense; tyrosol and its synthetic isomers 1,2- and 1,3-tyrosol were completely inactive on both bacteria . Among the derivatives of VW polyphenols considered, acetylcatechol and guaiacol were selectively toxic for Ps . savastanoi, while o-quinone was strongly toxic for both bacteria . The minor carboxylic polyphenols of VW at 10(-4) mol l-1 were all inactive on the bacteria . VW, catechol, 4-methylcatechol and the less abundant carboxylic polyphenols proved to be toxic on Hep2 human cells . Finally the possibility of using the active polyphenols in agriculture in an integrated pest management program for the protection of the olive plant is discussed.

J Bacteriol, 1995 Oct, 177(19), 5716 - 8
Identification of channel-forming activity in the cell wall of Corynebacterium glutamicum; Niederweis M et al.; The cell wall of the gram-positive Corynebacterium glutamicum was prepared . It contained an ion-permeable channel with a single-channel conductance of about 6 nS in 1 M KCl . The mobility sequence of the ions in the channel is similar to that in the aqueous phase, suggesting that it is a water-filled channel wide enough to allow unhindered diffusion of ions . The results indicate that we have identified the hydrophilic pathway through the mycolic acid layer of C . glutamicum.

Int J Syst Bacteriol, 1995 Oct, 45(4), 740 - 6
Phylogeny of the genus Corynebacterium deduced from analyses of small-subunit ribosomal DNA sequences; Ruimy R et al.; We determined almost complete small-subunit ribosomal DNA sequences of 50 reference strains belonging to the genera Corynebacterium, Rhodococcus, and Gordona and compared these sequences with previously published sequences . Three phylogenetic methods (the neighbor-joining, maximum-likelihood, and maximum-parsimony methods), as well as a bootstrap analysis, were used to assess the robustness of each topology which we obtained . The results of comparative phylogenetic analyses confirmed that the genera Corynebacterium, Dietzia, Gordona, Mycobacterium, Nocardia, Tsukamurella, and Turicella form a monophyletic taxon within the phylum containing the high-G+C-content gram-positive bacteria . The genus Corynebacterium appeared to be a monophyletic unit whose members could be divided into four major clusters . The validity of the genus Turicella is doubtful since members of this genus clearly belong to the genus Corynebacterium . The variability of chemotaxonomic characteristics within the genus Corynebacterium suggests that small-subunit ribosomal DNA sequence analysis is probably the most straightforward method for confirming that a bacterium belongs to this genus.

Int J Syst Bacteriol, 1995 Oct, 45(4), 735 - 9
Heterogeneity within human-derived centers for disease control and prevention (CDC) coryneform group ANF-1-like bacteria and description of Corynebacterium auris sp . nov; Funke G et al.; Recently, Centers for Disease Control and Prevention coryneform group ANF-1 bacteria were described as Corynebacterium afermentans, and group ANF-1-like bacteria were described as Turicella otitidis . Over a 1.5-year period 10 strains of a previously undescribed, gram-positive, rod-shaped organism that was not partially acid fast and resembled ANF-1-like bacteria were isolated from different pediatric patients with ear infections . These previously undescribed coryneform bacteria exhibited a distinct colony morphology and consistency, had a carbon source utilization pattern distinct from the carbon source utilization patterns of C . afermentans and T . otitidis, had a cell wall based on meso-diaminopimelic acid, contained mycolic acids, and had DNA G+C contents of 68 to 74 mol% . A 16S rRNA gene sequence analysis revealed that these clinical isolates are members of the genus Corynbacterium and that they are distinct from C . afermentans and T . otitidis . On the basis of phenotypic and phylogenetic evidence we propose a new species, Corynebacterium auris, for these Centers for Disease Control and Prevention coryneform group ANF-1-like bacteria . The type strain is strain DSM 44122 (CCUG 33426).

Int J Syst Bacteriol, 1995 Oct, 45(4), 724 - 8
Phylogenetic analysis of the genus Corynebacterium based on 16S rRNA gene sequences; Pascual C et al.; The 16S rRNA gene sequences of 30 strains representing 23 validated Corynebacterium species and 7 currently non-valid Corynebacterium species were determined . These sequences were aligned with the sequences of other Corynebacterium species and related actinomycete taxa . A comparative sequence analysis revealed that there is considerable phylogenetic depth and internal structure in the genus Corynebacterium . Turicella otitidis and the amycolate species Corynebacterium amycolatum were located at the periphery of the genus Corynebacterium . It was evident that the species of the genus Corynebacterium form a monophyletic association and, together with other chemotype IV and mycolic acid-containing taxa (including the genera Dietzia, Gordona, Mycobacterium, Nocardia, Rhodococcus, and Tsukamurella), form a natural suprageneric group.

Int J Syst Bacteriol, 1995 Oct, 45(4), 653 - 60
Phylogenetic analysis of mycolic acid-containing wall-chemotype IV actinomycetes and allied taxa by partial sequencing of ribosomal protein AT-L30; Ochi K; The phylogenetic relationships among 30 mycolic acid-containing wall chemotype IV actinomycete strains and 12 strains belonging to allied taxa were examined by determining the amino acid sequences of the ribosomal AT-L30 proteins of these organisms . Sequencing 20 N-terminal amino acids of AT-L30 preparations revealed that the members of the genera containing mycolic acid-containing actinomycetes form two clusters; the first cluster contains the genera Nocardia, Rhodococcus, Gordona, and Tsukamurella, and the second cluster contains the genera Corynebacterium and Mycobacterium . The genus Nocardia was placed in a clade containing the genus Rhodococcus . The data showed that Tsukamurella paurometabolum is closely related phylogenetically to the genus Gordona . The phylogenetic clusters identified were entirely consistent with the proposal of Goodfellow that the family Nocardiaceae should encompass the mycolate-containing, cell wall type IV actinomycete genera Nocardia, Rhodococcus, Gordona, and Tsukamurella . The genera Actinomyces and Micrococcus exhibited AT-L30 amino acid sequence characteristics intermediate between those of actinomycetes and those of typical eubacteria . The genera Nocardia, Gordona, Mycobacterium, Actinoplanes, and Micromonospora were each a taxon that consisted of phylogenetically coherent species . In contrast, the genera Rhodococcus and Corynebacterium are taxa that consist of phylogenetically distantly related species . In general, my results are consistent with previous 16S rRNA sequencing results, but significant differences were also found . My data, together with previous AT-L30 sequencing data, show that phylogenetic relationships among taxa can be determined by using markers other than the ribosomal gene sequences.

J Infect, 1995 Sep, 31(2), 153 - 7
Epidemiology and molecular characterisation of toxigenic Corynebacterium diphtheriae var mitis from a case of cutaneous diphtheria in Manchester; Hamour AA et al.; Diphtheria is now an uncommon disease in Britain . We describe an imported case of cutaneous diphtheria in a previously immunised adult cause by C . diphtheriae var mitis . The control measures adopted to deal with the index case and two secondary cases so as to limit further spread among household and school contacts are outlined . Molecular typing was used to study the mode of spread of the organism among contacts.

J S Afr Vet Assoc, 1995 Sep, 66(3), 160 - 9
A comparative microbiological study of clinically healthy eyes and those affected by ophthalmia in cattle and the association of noctuid eye-frequenting moths; Gouws JJ et al.; The eyes of clinically healthy Simmentaler cattle and those affected by ophthalmia were sampled once a month over a continuous period of 12 months for bacterial, mycoplasmal and ureaplasmal infections . In total 478 eyes, representing from a clinical viewpoint 414 healthy and 64 affected eyes, were swabbed . Bacteria were isolated from 201 (48.6%) healthy eyes and 56 (87.5%) affected eyes . No bacteria were isolated from the remaining eyes . Eleven genera of bacteria were isolated from healthy eyes and 8 genera from affected eyes . The majority of isolates were classified in the genera Moraxella, Neisseria and Staphylococcus . Mycoplasmas were isolated from 247 (50.7%) healthy eyes and 27 (42.2%) affected eyes . No mycoplasmas were isolated from the remaining eyes . Ureaplasmas were not isolated from any animal . Eye-frequenting moths were collected on 3 occasions during the investigation and bacterial and mycoplasmal isolation techniques were performed on a total of 21 moths . Twelve different genera of bacteria, mostly Nocardia, Corynebacterium, Staphylococcus, Moraxella, and mycoplasmas were isolated from various eye-frequenting moths . Scanning electron microscopical studies of the proboscis of the moths showed it to contain various sensillae and short triangular denticles that could possibly cause damage to the mucous membranes of the eyes and predispose to ophthalmia in cattle.

Int J Immunopharmacol, 1995 Sep, 17(9), 779 - 86
Stimulation of macrophages with IFN gamma or TNF alpha shuts off the suppressive effect played by PGE2; Zicari A et al.; PGE2 has been shown to be able to interfere with various lymphocyte and macrophage functions, but its effects on macrophage activation are still unclear . In this study, carried out on peritoneal macrophages obtained from healthy, tumour-bearing and Corynebacterium parvum-treated mice, we demonstrated that PGE2 is involved in the down-regulation of macrophage activation, but it cannot exert its inhibiting effect when macrophages are further stimulated with activating cytokines, such as IFN gamma and TNF alpha . Our findings provide new insight into how macrophage tumoricidal activity may be induced and maintained even in presence of significant levels of PGE2.

AJR Am J Roentgenol, 1995 Sep, 165(3), 669 - 71
Gadolinium-based MR contrast media: potential for growth of microbial contaminants when single vials are used for multiple patients; Green KA et al.; OBJECTIVE . To assess potential risk if single-dose vials of gadolinium-based contrast media are used for multiple patients . The use of multidose vials can pose a significant risk if contaminants are accidentally introduced into the vials and proliferate . Therefore, we tested the ability of gadolinium-based contrast to support the growth of a variety of microbial pathogens . MATERIALS AND METHODS . We collected the unused portions of 15 single-use vials of gadolinium-based contrast media . The residual contrast material was refrigerated after use and checked for sterility prior to inoculation with a selection of gram-positive and gram-negative bacteria and yeast . Contrast material was incubated at room temperature and at 4 degrees C, and quantitative cultures were performed at 0, 24, 48, and 72 hr and at 7 days . RESULTS . No microbial growth occurred from cultures of the used contrast solutions prior to inoculation . None of the organisms tested in our study proliferated in the contrast material . All test organisms persisted at least 48 hr after inoculation at both temperatures . At 7 days, Staphylococcus aureus, Streptococcus epidermidis, and Corynebacterium jeikeium were recovered in significant quantities . Colony counts of Serratia odorifera rapidly decreased at room temperature but persisted beyond 7 days at 4 degrees C . CONCLUSION . The risk of contaminating vials of contrast agents punctured aseptically is small . Our study demonstrates a lack of proliferation of organisms in the two gadolinium-based compounds tested, suggesting that these solutions could be used for more than one procedure.

Infect Immun, 1995 Sep, 63(9), 3559 - 66
Leishmania pifanoi amastigote antigens protect mice against cutaneous leishmaniasis; Soong L et al.; In the search for a leishmaniasis vaccine, extensive studies have been carried out with promastigote (insect stage) molecules . Information in this regard on amastigote (mammalian host stage) molecules is limited . To investigate host immune responses to Leishmania amastigote antigens, we purified three stage-specific antigens (A2, P4, and P8) from in vitro-cultivated amastigotes of Leishmania pifanoi by using immunoaffinity chromatography . We found that with Corynebacterium parvum as an adjuvant, three intraperitoneal injections of 5 micrograms of P4 or P8 antigen provided partial to complete protection of BALB/c mice challenged with 10(5) to 10(7) L . pifanoi promastigotes . These immunized mice developed significantly smaller or no lesions and exhibited a 39- to 1.6 x 10(5)-fold reduction of lesion parasite burden after 15 to 20 weeks of infection . In addition, P8 immunization resulted in complete protection against L . amazonensis infection of CBA/J mice and partial protection of BALB/c mice, suggesting that this antigen provided cross-species protection of mice with different H-2 haplotypes . At different stages during infection, vaccinated mice exhibited profound proliferative responses to parasite antigens and increased levels of gamma interferon production, suggesting that a Th1 cell-mediated immune response is associated with the resistance in these mice . Taken together, the data in this report indicate the vaccine potential of amastigote-derived antigens.

Dig Dis Sci, 1995 Sep, 40(9), 2070 - 3
Conventional dose of omeprazole alters gastric flora; Karmeli Y et al.; Quantitative cultures were carried out on samples from gastric juice obtained from 12 ambulatory patients with esophagitis before and one month after omeprazole therapy . An increase in the number of patients in whom gastric juice was culture-positive, as well as an increment in the bacterial counts were noted . The spectrum of microorganisms isolated from gastric juice was identical to the normal flora of the oral cavity, mainly alpha-hemolytic streptococci, corynebacteria, and Candida species . Thus, the counts of organisms within gastric contents are simply a reflection of swallowed oral microflora that were able to survive due to the less acidic environment.

J Clin Microbiol, 1995 Sep, 33(9), 2458 - 61
Phenotypic characteristics of 31 strains of Corynebacterium striatum isolated from clinical samples; Martinez-Martinez L et al.; During a 34-month period (January 1991 to October 1993), 31 Corynebacterium striatum stains recovered from clinical samples from 24 patients were characterized . Twenty (64%) strains were isolated from wound exudates, 5 (16%) were isolated from bronchial aspirates, 2 (7%) were isolated from urine, 2 (7%) were isolated from endotracheal tubes, 1 (3%) was isolated from a catheter, and 1 (3%) was isolated from empyema . The organisms were identified by conventional culture and phenotypic characterization, the API CORYNE system, and cellular fatty acid composition analyses . The colonies of C . striatum could be confused with those of coagulase-negative staphylococci upon primary isolation from clinical material . A consistent phenotypic pattern was observed: all strains reduced nitrate, hydrolyzed tyrosine, and produced acid from glucose, fructose, and sucrose but not from maltose . API CORYNE profile numbers were 3100105 (28 strains) and 3000105 (3 strains) . Susceptibility testing of C . striatum was performed by disk diffusion . All strains were susceptible to both imipenem and vancomycin and resistant to fosfomycin; most strains were susceptible to ampicillin and cephalosporins and resistant to clindamycin, erythromycin, and tetracycline . Performing a Gram stain of fosfomycin-resistant "Staphylococcus-like" colonies was critical in order to identify C . striatum.

J Clin Microbiol, 1995 Sep, 33(9), 2244 - 9
Corynebacterium seminale sp . nov., a new species associated with genital infections in male patients; Riegel P et al.; We studied 12 coryneform isolates having similar biochemical profiles which did not permit their assignment to any recognized taxa . Human semen was the source for seven of these strains, whereas the other strains were isolated from urethra, urine, and blood specimens of adult male patients . These bacteria were found in significant quantities (10(4) to 10(5) CFU/ml) in semen specimens from infertile male patients with the diagnosis of prostatitis . These strains had characteristics of the genus Corynebacterium, such as 60 mol% G + C in the DNA and corynemycolic acids, meso-diaminopimelic acid, arabinose, and galactose in the cell wall . Quantitative DNA-DNA hybridizations (S1 nuclease procedure) and phylogenies based on comparisons of almost-complete small-subunit ribosomal DNA sequences confirmed that these strains constitute a single new species within the genus Corynebacterium . All 12 strains showed similar phenotypic features, i.e., good growth on sheep blood agar in contrast with poor growth on the same medium supplemented with 1% Tween 80, a positive CAMP test in the presence of Staphylococcus aureus, glucose and sucrose fermentation, and the presence of beta-glucuronidase . Some strains reduced nitrate and hydrolyzed urea or esculin . These features allowed us to distinguish these strains from members of any other coryneform taxon, and the proposed name is Corynebacterium seminale with strain IBS B12915 (CIP 104297) as the type strain . The description and delineation of these strains as a new species should be useful for further studies, including evaluations of their prevalence among the normal flora and their clinical implications.

J Biol Chem, 1995 Aug 4, 270(31), 18252 - 9
Sequence analysis of sarcosine oxidase and nearby genes reveals homologies with key enzymes of folate one-carbon metabolism; Chlumsky LJ et al.; Corynebacterial sarcosine oxidase, a heterotetrameric (alpha beta gamma delta) enzyme containing covalent and noncovalent FAD, catalyzes the oxidative demethylation of sarcosine to yield glycine, H2O2, and 5,10-CH2-tetrahydrofolate (H4folate) in a reaction requiring H4folate and O2 . The sarcosine oxidase operon contains at least five closely packed genes encoding sarcosine oxidase subunits and serine hydroxymethyltransferase (glyA), arranged in the order glyAsoxBDAG . The operon status of a putative purU gene, found 340 nucleotides downstream from soxG, is not known . No homology with other proteins is observed for the smallest sarcosine oxidase subunits gamma and delta . The beta subunit (405 residues) contains an ADP-binding motif near its NH2 terminus, the covalent FAD attachment site (H175), and exhibits homology with the NH2-terminal half of dimethylglycine dehydrogenase (857 residues) and monomeric, bacterial sarcosine oxidases (approximately 388 residues), enzymes that contain a single covalent FAD . The alpha subunit (967 residues) contains a second ADP-binding motif within an approximately 280 residue region near the NH2 terminus that exhibits homology with subunit A from octopine and nopaline oxidases, heterodimeric enzymes that catalyze analogous oxidative cleavage reactions with N-substituted arginine derivatives . An approximately 380 residue region near the COOH terminus of alpha exhibits homology with T-protein and the COOH-terminal half of dimethylglycine dehydrogenase . These enzymes catalyze the formation of 5,10-CH2-H4folate, using different one-carbon donors . The results suggest that the alpha subunit and dimethylglycine dehydrogenase contain an NH2-terminal domain that binds noncovalent or covalent FAD, respectively, and a carboxyl-terminal H4folate-binding domain.

J Vet Med Sci, 1995 Aug, 57(4), 715 - 9
Pathogenicity of Corynebacterium kutscheri in the Syrian hamster; Amao H et al.; The pathogenicity of Corynebacterium kutscheri isolated for the first time from Syrian hamster was experimentally studied in hamsters . In hamsters given intramuscular (i.m.) or subcutaneous (s.c.) inoculation with 10 or 10(3) bacteria, neither clinical signs nor gross lesions were found . In those given 10(5) bacteria i.m., moderate proliferation of granulation tissue was found in the muscle of the inoculation region at necropsy . In the animals given 10(5) bacteria s.c., a nodular lesion was observed at the inoculation site 2 days post-inoculation (p.i.), but the nodules subsided gradually from 6 days p.i . and were unclear 10 days p.i . At necropsy, small abscesses were found in all the animals in this group . In those given 10(7) bacteria either i.m . or s.c., lesions were clearly observed at the inoculation site 1 to 10 days p.i., and a large abscess was noted at necropsy . The organisms were isolated only from the lesions in the groups . Agglutinating antibody in the sera was detected only in the animals given 10(5) or 10(7) bacteria . This suggests that 10(5) of C . kutscheri are needed to form localized nodular abscesses in Syrian hamsters.

J Bacteriol, 1995 Aug, 177(16), 4690 - 5
Glycine betaine uptake after hyperosmotic shift in Corynebacterium glutamicum; Farwick M et al.; Osmoregulatory uptake of glycine betaine in whole cells of Corynebacterium glutamicum ATCC 13032 (wild type) was studied . The cells actively take up glycine betaine when they are osmotically shocked . The total accumulation and uptake rate were dependent on the osmotic strength of the medium . Kinetic analysis revealed a high-affinity transport system (Km, 8.6 +/- 0.4 microM) with high maximum velocity (110 nmol.min-1.mg {dry weight}-1) . Glycine betaine functioned as a compatible solute when added to the medium and allowed growth at an otherwise inhibitory osmotic strength of 1.5 M NaCl . Proline and ectoine could also be used as osmoprotectants . Glycine betaine is neither synthesized nor metabolized by C . glutamicum . The glycine betaine transport system is constitutively expressed at a basal level of activity . It can be induced up to eightfold by osmotic stress and is strongly regulated at the level of activity . The transport system is highly specific and has its pH optimum in the slightly alkaline range at about pH 8 . The uptake of the zwitterionic glycine betaine is mediated by a secondary symport system coupled to cotransport of at least two Na+ ions . It is thus driven both by the membrane potential and the Na+ gradient . An extremely high accumulation (internal/external) ratio of up to 4 x 10(6) was measured, which represents the highest accumulation ratio observed for any transport system.

Biochem J, 1995 Aug 1, 309 ( Pt 3), 999 - 1007
The biosynthesis of threonine by mammalian cells: expression of a complete bacterial biosynthetic pathway in an animal cell; Rees WD et al.; The coding regions for the Escherichia coli gene for aspartokinase I/homoserine dehydrogenase I (thrA) and the Corynebacterium glutamicum gene for aspartic semialdehyde dehydrogenase (asd) have been subcloned into a Simian Virus 40 (SV40)-based mammalian expression vector . Both enzyme activities are expressed in mouse 3T3 cells after transfer of the corresponding chimaeric gene . The kinetic parameters are similar to those of the native bacterial enzymes, and aspartokinase I/homoserine dehydrogenase I retains its allosteric regulation by threonine . An extract of the cells expressing aspartokinase I/homoserine dehydrogenase I, mixed with one from cells expressing aspartic semialdehyde dehydrogenase, produced homoserine when the mixture was incubated with aspartic acid, ATP and NADPH . The thrA and asd expression cassettes were combined into a single plasmid which, when transfected into 3T3 cells, enabled them to produce homoserine from aspartic acid . Homoserine-producing 3T3 cells were transfected with the plasmid pSVthrB/C (homoserine kinase and threonine synthase) and selected for growth on homoserine . Cell lines isolated from these cells expressed the complete bacterial threonine pathway, were independent of threonine for growth and could be maintained in medium which contained no free threonine . The threonine in the proteins of these cells became enriched in 15N when the culture medium contained {15N}aspartic acid . The production of homoserine and the growth of cells was at a maximum when there was more than 2.5 mM aspartate in the medium . Below this concentration the high Km of aspartokinase limited the flux through the pathway . In the presence of additional aspartic acid the new pathway could sustain a cell cycle time close to that of the same cells cultured in threonine-containing medium.

Am J Respir Cell Mol Biol, 1995 Aug, 13(2), 185 - 95
The role of tumor necrosis factor in increased airspace epithelial permeability in acute lung inflammation; Li XY et al.; Increased airspace epithelial permeability is an early event in lung inflammation and injury . In this study, we have developed a rat model to study the mechanisms of the epithelial permeability to 125iodine-labeled bovine serum albumin (125I-BSA), instilled intratracheally during acute lung inflammation . Epithelial permeability was measured as the percentage of instilled 125I-BSA appearing in the blood . The increase in epithelial permeability induced by intratracheal instillation of heat-killed Corynebacterium parvum produced a peak influx of neutrophils into the bronchoalveolar space at 16 h, which occurred after the peak increase in epithelial permeability (8 h) . The increased epithelial permeability induced by C . parvum did not appear to be protease- or oxidant-mediated . Depletion of peripheral blood neutrophils was achieved by an intravenous injection of anti-neutrophil polyclonal antibody . The consequent profound reduction in neutrophil and macrophage influx into the airspaces 8 h after instillation of C . parvum reduced the epithelial permeability to control values . Bronchoalveolar lavage (BAL) leukocytes from rats 8 h, but not 16 h, after treatment with C . parvum caused a modest increase in epithelial permeability when re-instilled intratracheally into control rat lungs . Separation of the leukocytes before re-instillation indicated that macrophages rather than neutrophils were predominantly responsible for the increased epithelial permeability . The presence of dramatically increased levels of tumor necrosis factor (TNF) in BAL 8 h in contrast to a slight increase in BAL 16 h after C . parvum, the release of TNF from 8 h macrophages, the increased epithelial permeability induced by TNF in epithelial monolayers in vitro, and the inhibition of C . parvum-induced epithelial permeability by TNF antibody support the premise that TNF is a major player in the increased epithelial permeability that occurs during C . parvum-induced acute alveolitis.

Eur J Immunol, 1995 Aug, 25(8), 2325 - 31
Tumor cells cotransfected with interleukin-7 and B7.1 genes induce CD25 and CD28 on tumor-infiltrating T lymphocytes and are strong vaccines; Cayeux S et al.; Interleukin-7 (IL-7) and the membrane molecule B7 are both able to provide proliferation and activation signals for T cells . However, tumor cells transfected to express either molecule alone are not reliably rejected in syngeneic hosts or are not sufficiently immunogenic to serve as potent tumor vaccines . Since IL-7 and B7 have shown synergistically to induce activation and proliferation of T cells in vitro, we have expressed B7.1 by means of a retrovirus in the mammary adenocarcinoma TS/A which arose spontaneously in a BALB/c mouse and in the plasmacytoma J558L and their IL-7-transfected sublines to improve vaccine efficacy . Expression of IL-7 or B7.1 alone in tumor cells decreased tumorigenicity, but nevertheless tumors grew in a substantial number of mice . In contrast, IL-7/B7.1 cotransfected cells did not grow as tumor in a single case . This inhibition of tumor growth was completely T cell dependent, because TS/A-IL-7/B7.1 cells retained their full tumorigenic potential in T cell-deficient mice . Analysis of tumor-infiltrating T lymphocytes revealed increased numbers of T cells in B7, IL-7 and IL-7/B7 transfected compared to parental tumors . In IL-7/B7 transfected tumors, T cell numbers were not further increased compared to that in single-gene-transfected tumors . However, T cells in B7 and IL-7 transfected tumors differed phenotypically with respect to activation markers . In B7 transfected tumors, T cells were predominantly CD28+ and CD25-, while in IL-7 transfected tumors, T cells were mainly CD28- and CD25+ . In IL-7/B7 cotransfected tumors, the majority of T cells was CD28+ and CD25+ . Thus, IL-7 and B7 induced an anti-tumor immune response by complementary T cell directed pathways in a cooperative fashion . Importantly, immunization of mice with the transfected cells and subsequent contralateral challenge with parental tumor cells showed that IL-7/B7 co-expressing cells induced the most strongly protective immunity, which is superior to that induced by single-gene transfectants and to the adjuvant Corynebacterium parvum . Vaccine efficacy was abrogated when irradiated cells were used for vaccination . Together, our results show that IL-7 and B7.1 transfected tumor cells induce strong T cell activation and tumor immunity.

Lab Anim Sci, 1995 Aug, 45(4), 366 - 7
Acute pneumonia in a Syrian hamster: isolation of a Corynebacterium species; Tansey G et al.; A respiratory tract illness was detected in a 1-year-old male Syrian hamster; after it failed to respond to antibiotic therapy, the hamster was euthanized by CO2 administration . Postmortem examination revealed acute edematous pneumonia, and Corynebacterium paulometabulum was isolated from the lungs.

Proc Natl Acad Sci U S A, 1995 Jul 18, 92(15), 6803 - 7
Transition metal ion activation of DNA binding by the diphtheria tox repressor requires the formation of stable homodimers; Tao X et al.; The diphtheria tox repressor (DtxR) is a transition metal ion-dependent regulatory element that controls the expression of diphtheria toxin and several genes involved in the synthesis of siderophores in Corynebacterium diphtheriae . In the presence of transition metal ions apo-DtxR becomes activated and specifically binds to its target DNA sequences . We demonstrate by glutaraldehyde cross-linking that monomeric apo-DtxR is in weak equilibrium with a dimeric form and that upon addition of activating metal ions to the reaction mixture a dimeric complex is stabilized . Addition of the DNA-binding-defective mutant apo-DtxR(delta 1-47) to apo-DtxR in the absence of transition metal ions inhibits conversion of the apo-repressor to its activated DNA-binding form . We also show that the binding of Ni2+ to both apo-DtxR and apo-DtxR(delta 1-47) is cooperative and that upon ion binding there is a conformational change in the environment of the indole ring moiety of Trp-104 . For the wild-type repressor the consequences of this conformational change include a shift in equilibrium toward dimer formation and activation of target DNA binding by the repressor . We conclude that the formation of DtxR homodimers is mediated through a protein-protein interaction domain that is also activated on metal ion binding.

Rev Inst Med Trop Sao Paulo, 1995 Jul-Aug, 37(4), 291 - 6
Genetic relationships of Corynebacterium diphtheriae strains isolated from a diphtheria case and carriers by restriction fragment length polymorphism of rRNA genes; Sacchi CT et al.; In the present study we report the results of an analysis, based on ribotyping of Corynebacterium diphtheriae intermedius strains isolated from a 9 years old child with clinical diphtheria and his 5 contacts . Quantitative analysis of RFLPs of rRNA was used to determine relatedness of these 7 C.diphtheriae strains providing support data in the diphtheria epidemiology . We have also tested those strains for toxigenicity in vitro by using the Elek's gel diffusion method and in vivo by using cell culture method on cultured monkey kidney cell (VERO cells) . The hybridization results revealed that the 5 C.diphtheriae strains isolated from contacts and one isolated from the clinical case (nose case strain) had identical RFLP patterns with all 4 restriction endonucleases used, ribotype B . The genetic distance from this ribotype and ribotype A (throat case strain), that we initially assumed to be responsible for the illness of the patient, was of 0.450 showing poor genetic correlation among these two ribotypes . We found no significant differences concerned to the toxin production by using the cell culture method . In conclusion, the use of RFLPs of rRNA gene was successful in detecting minor differences in closely related toxigenic C.diphtheriae intermedius strains and providing information about genetic relationships among them.

Int J Syst Bacteriol, 1995 Jul, 45(3), 533 - 7
Corynebacterium argentoratense sp . nov., from the human throat; Riegel P et al.; A new Corynebacterium species, Corynebacterium argentoratense was isolated from the throats of four human patients . It is characterized by the presence of chemotype IV, a cell wall, corynomycolic acids, and a G+C content ranging from 60 to 61 mol% . Strains belonging to this species exhibit high levels of DNA relatedness as determined by DNA-DNA hybridization experiments (S1 nuclease procedure) but no close DNA relatedness with related Corynebacterium species . Phylogenies based on comparative analyses of nearly complete small-subunit rDNA sequences confirmed the inclusion of this new species within the genus Corynebacterium and grouped it in a cluster with C . diphtheriae, C . ulcerans, C . pseudotuberculosis, and C . kutscheri . PCR experiments revealed an absence of the gene coding for diphtheria toxin . This new species can be identified by its mycolic acid pattern, fermentation of sugars, and enzymatic activities . Strain IBS B10697 (CIP 104296) is the type strain of C . argentoratense.

Indian J Lepr, 1995 Jul-Sep, 67(3), 309 - 19
Studies on microbial aerobic flora of skin in leprosy patients; Sharma RK et al.; This study reports the isolation and identification of aerobic organisms from biopsies/slit-skin smears/scrapings from 129 leprosy patients and 50 healthy controls . These include 56 paucibacillary (PB) and 73 multibacillary (MB) cases . Thirty-six isolates from the specimens from 21 patients and 15 healthy controls were grown . The non-mycobacterial isolates from clinically PB leprosy (TT/BT/I) patients were: (1) Gram-positive cocci: Staphylococcus aureus(1), Staphylococcus albus(1); (b) Gram-positive bacilli: Bacillus subtilis(1), Corynebacterium xerosis(1); (c) Gram-negative bacilli: Escherichia coli(1), Proteus mirabilis(2), Klebsiella pneumoniae(1) and Pseudomonas aeruginosa(1) . The isolates from clinically MB leprosy (BB/BL/LL) patients were: (a) Gram-positive cocci: Micrococci(1), Staphylococcus aureus(1) and Staphylococcus albus(1); (b) Gram-positive bacilli: Corynebacterium xerosis(1); Corynebacterium hofmanni(1) and Bacillus cereus(1) . (c) Gram-negative bacilli: Escherichia coli(2), Klebsiella pneumoniae(1) and Proteus mirabilis(2) . The specimens from healthy controls yielded similar organisms . These were (a) Gram-positive cocci: Staphylococcus albus(2), Staphylococcus aureus(2) and Micrococci(2); (b) Gram-positive bacilli: Corynebacterium xerosis(1), Bacillus subtilis(2), Corynebacterium hofmanni(1) and Bacillus cereus(1); (c) Gram-negative bacilli: Escherichia coli(3), Proteus vulgaris(1) and Proteus mirabilis(1) . While these results show no significant differences in the species types of non-mycobacterial aerobic organisms isolated from healthy skin and PB/MB types of leprosy, these isolates need to be characterized by immunological/molecular methods to find out subtypes if any.

Mikrobiologiia, 1995 Jul-Aug, 64(4), 437 - 41
{Relationship between the synthesis of a new stress response component and bacterial cell metabolism}; Lysak EI et al.; Various stress factors (incubation with redox-cycling agents, ozonization, heat shock) that induced accumulation of 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEC) were also shown to induce various alterations of the phospholipid composition in three microbial species: Corynebacterium ammoniagenes, Micrococcus luteus, and Staphylococcus aureus . The influence of adding 10 carbohydrates to the growth medium on MEC accumulation by C . ammoniagenes cells was tested . Only glucose and mannose exerted a pronounced stimulatory effect on the MEC synthesis under oxidative stress . The target of oxidative attack whose transformation causes MEC synthesis is suggested to be located near or on the bacterial cytoplasmic membrane.

Aust Vet J, 1995 Jul, 72(7), 266 - 9
The spread of Corynebacterium pseudotuberculosis infection to unvaccinated and vaccinated sheep; Paton MW et al.; The decrease in the prevalence of Corynebacterium pseudotuberculosis after two generations of vaccination against the disease it causes, was used to estimate the rate of control of caseous lymphadenitis (CLA) . Three groups of 150 sheep, of which 50 in each group were artificially infected with C pseudotuberculosis and 100 in each group were uninfected sheep, were run separately for 40 months and shorn 5 times to promote the spread of CLA . One lot of 50 infected sheep and 2 lots of 100 uninfected sheep were vaccinated against CLA . The rate of spread of CLA was recorded . Sheep vaccinated against CLA and naturally exposed to infection had a 74% lower infection rate than unvaccinated sheep . Sheep vaccinated against CLA and exposed to only vaccinated infected sheep had a 97% lower infection rate . Unvaccinated sheep had a 76% infection rate, with 77% of the transmission occurring at the 4th and 5th shearings, without any discharging CLA abscesses being observed . This study supports the view that in Australian wool producing flocks, CLA spreads mainly from sheep with discharging lung abscesses to sheep with shearing cuts . Vaccinated sheep infected with CLA have 96% fewer lung abscesses compared with unvaccinated infected sheep and are therefore less likely to spread this disease to other sheep.

J Infect, 1995 Jul, 31(1), 63 - 5
An atypical case of Corynebacterium diphtheriae endocarditis and subsequent outbreak control measures; Booth LV et al.; An atypical case of Corynebacterium diphtheriae endocarditis with severe rhabdomyolysis and cerebral emboli is presented . The patient underwent successful mitral and aortic valve replacements and is only the third reported case with a successful outcome following surgery . Outbreak control measures were complicated by an equivocal result from guinea pig toxin tests.

Klin Lab Diagn, 1995 Jul-Aug, (4), 45 - 8
{Corynebacteria isolated in colpitis and puerperal complications}; Martikainen ZM; The role of microorganisms belonging to the genus Corynebacterium in normalization of vaginal biocenosis by creating acid medium is discussed . A total of 300 samples of lochia and vaginal and cervical secretion were examined, 140 of these taken from women without gynecological diseases and women with a normal course of the postpartum period (controls) and 160 from patients with colpitis of various origins and a complicated course of the puerperium . An appreciable increase of the level of diphtheroids was observed in the lochia of control subjects and in all samples of patients (p < 0.01) . Eight species of Corynebacterium were isolated . Urease-negative C . minutissimum, C . equi, C . aquaticum, and C . xerosis predominated in both controls and patients (p < 0.01) . Opportunistic C . bovis, C . enzymicum, C . kutshevi, and C . sp . possessing urease activity were seldom isolated . No differences between the 2 groups in the species composition of the isolated bacteria were detected (p > 0.05) . Two species of Corynebacterium were sometimes isolated from the same sample, this being more frequently with vaginal and cervical secretion samples than with lochia both in controls and patients (p < 0.01) . The most incident association was C . aquaticum and C . equi . 59.6 +/- 2.7 isolated strains proved to be sensitive to antibiotics manufactured in this country.

Appl Microbiol Biotechnol, 1995 Jul, 43(3), 482 - 8
Production of isoleucine by overexpression of ilvA in a Corynebacterium lactofermentum threonine producer; Colon GE et al.; Overproduction of isoleucine, an essential amino acid, was achieved by amplification of the gene encoding threonine dehydratase, the first enzyme in the threonine to isoleucine pathway, in a Corynebacterium lactofermentum threonine producer . Threonine overproduction was previously achieved with C . lactofermentum ATCC 21799, a lysine-hyperproducing strain, by introduction of plasmid pGC42 containing the Corynebacterium homdr and thrB genes (encoding homoserine dehydrogenase and homoserine kinase respectively) under separate promoters . The pGC42 derivative, pGC77, also contains ilvA, which encodes threonine dehydratase . In a shake-flask fermentation, strain 21799(pGC77) produced 15 g/l isoleucine, along with small amounts of lysine and glycine . A molar carbon balance indicates that most of the carbon previously converted to threonine, lysine, glycine and isolecine was incorporated into isoleucine by the new strain . Thus, in our system, simple overexpression of wild-type ilvA sufficed to overcome the effects of feedback inhibition of threonine dehydratase by the end-product, isoleucine.

J Bacteriol, 1995 Jul, 177(14), 4021 - 7
Unbalance of L-lysine flux in Corynebacterium glutamicum and its use for the isolation of excretion-defective mutants; Vrljic M et al.; We found that the simple addition of L-methionine to the wild type of Corynebacterium glutamicum results in excretion of the cellular building block L-lysine up to rates of 2.5 nmol/min/mg (dry weight) . Biochemical analyses revealed that L-methionine represses the homoserine dehydrogenase activity and reduces the intracellular L-threonine level from 7 to less than 2 mM . Since L-lysine synthesis is regulated mainly by L-threonine (plus L-lysine) availability, the result is enhanced flux towards L-lysine . This indicates a delicate and not well controlled type of flux control at the branch point of aspartate semialdehyde conversion to either L-lysine or L-threonine, probably due to the absence of isoenzymes in C . glutamicum . The inducible system of L-lysine excretion discovered was used to isolate mutants defective in the excretion of this amino acid . One such mutant characterized in detail accumulated 174 mM L-lysine in its cytosol without extracellular excretion of L-lysine, whereas the wild type accumulated 53 mM L-lysine in the cytosol and 5.9 mM L-lysine in the medium . The mutant was unaffected in L-lysine uptake or L-isoleucine or L-glutamate excretion, and also the membrane potential was unaltered . This mutant therefore represents a strain with a defect in an excretion system for the primary metabolite L-lysine.

J Vet Diagn Invest, 1995 Jul, 7(3), 347 - 51
Evaluation of an enzyme-linked immunosorbent assay licensed by the USDA for use in cattle for diagnosis of ovine paratuberculosis; Dubash K et al.; A commercially available Mycobacterium phlei-absorbed enzyme-linked immunosorbent assay (ELISA) approved to detect antibodies to Mycobacterium paratuberculosis in cattle was evaluated for its applicability in sheep . The potential for interference with ELISA results from cross-reacting antibodies to Corynebacterium pseudotuberculosis was also investigated . Serum samples were randomly selected from a collection of samples obtained in 1986-1991 from 6 infected and 5 noninfected sheep flocks varying in breed, age, and geographic origin . Tests were performed on sera from 27 paratuberculous sheep, confirmed by histopathology, bacteriologic culture, and/or acid-fast staining of ileal mucosal smears, and on sera from 246 noninfected sheep . The optical density of each sample was expressed as a percentage of the optical density of a known positive sheep serum sample tested on the same plate . These values were log-transformed to achieve normality of distribution, and sensitivity and specificity estimates were calculated based on 2 and 3 standard deviations above the mean of the percent positive value (PPV) of the noninfected sheep . A cutoff value of PPV > or = 55.74 resulted in an estimated sensitivity of 0.48 and a specificity of 0.95 . Sera from 10 noninfected sheep with PPV above the cutoff level of 55.74% were absorbed with heat-treated C . pseudotuberculosis organisms in addition to M . phlei antigens . Sera from 14 ELISA-positive paratuberculous sheep and 23 ELISA-negative noninfected sheep were similarly treated, and results were compared . Absorption with C . pseudotuberculosis resulted in a significant decrease in PPV in all 3 groups of sheep sera, but a greater decrease was observed in the noninfected sheep with PPV above the cutoff level when compared with noninfected sheep with PPV below that level . Results of this study suggest that ELISA may be of value in screening sheep flocks for paratuberculosis, but further experimentation is needed to optimize the sensitivity and specificity of the assay . Exposure to C . pseudotuberculosis may confound results obtained by M . phlei-absorbed ELISA for paratuberculosis.

Lab Anim, 1995 Jul, 29(3), 294 - 9
Serological studies of Corynebacterium kutscheri and coryneform bacteria using an enzyme-linked immunosorbent assay (ELISA); Boot R et al.; An enzyme-linked immunosorbent assay (ELISA) to measure Corynebacterium kutscheri antibodies in mice and rats was developed . Seven C . kutscheri isolates showed considerable serological relationship, but Japanese isolates differed from the British isolates . The ELISA appeared specific since C . kutscheri antigen did not react with antisera against 8 heterologous coryneform species . Antibodies to C . kutscheri were to a limited extent absorbed by autologous and homologous antigen, but not at all by the heterologous coryneform species . In naturally infected wild Rattus norvegicus and laboratory NA rats, the ELISA demonstrated high ODs to C . kutscheri.

Z Gastroenterol, 1995 Jul, 33(6), 362 - 7
{Masked course of Whipple disease with uveitis, infection, endocardial involvement and abdominal lymphomas--case report and review of the literature}; Hollerbach S et al.; Whipple's disease is a systemic disease which may virtually affect any organ system, but in many cases it involves the small intestine causing gastrointestinal symptoms . The differential diagnosis is difficult since symptoms may be nonspecific . We report the case of a 44-year old white male patient with a history of migrating arthralgia and chronic fatigue . The patient newly developed an uveitis and underwent a vitrectomy; the further clinical work-up including gastroscopy with intestinal biopsy revealed no sufficient diagnosis . Subsequently, the patient's condition deteriorated with marked weight loss, fever and progressive weakness . An anaerobic sepsis with a corynebacterium was confirmed and with i.v.-antibiotics the patients's condition improved markedly . The further examinations disclosed enlarged mesenteric lymph nodes and the involvement of other organs (endocard, liver) . CT-guided biopsy only showed fatty degeneration, but operative adenectomy confirmed Whipple's disease . The patient remained without relapse on long-term antibiotic treatment with doxycycline until today . Obviously, in our case the intestinal biopsies failed to detect Whipple's disease after the successful initiation of antibiotic treatment . In the absence of gastrointestinal findings and with concomitant secondary diseases the definitive diagnosis can be difficult . In addition, the previous uveitis and the endocardial involvement are most interesting.

J Bacteriol, 1995 Jun, 177(12), 3512 - 7
Cloning of a Corynebacterium diphtheriae iron-repressible gene that shares sequence homology with the AhpC subunit of alkyl hydroperoxide reductase of Salmonella typhimurium; Tai SS et al.; To understand how Corynebacterium diphtheriae responds to iron limitation, we compared the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) protein profiles of both wild-type cells and iron uptake mutants grown in either high- or low-iron medium . The removal of iron by ethylene diamine di-(o-hydroxy-phenyl acetic acid) from the growth medium of wild-type cells resulted in induction of at least 14 polypeptides . DirA, a major iron-repressible polypeptide, was purified from wild-type cells by preparative SDS-PAGE, and the dirA structural gene was isolated from a genomic library of nontoxigenic C . diphtheriae . The nucleotide sequence of dirA was determined, and the deduced amino acid sequence of DirA revealed strong homologies with the AhpC subunit of Salmonella typhimurium alkyl hydroperoxide reductase and polypeptides of other microorganisms associated with oxidation reduction activity . Like AhpC, cloned DirA reduced the susceptibility of an Escherichia coli ahp mutant to cumene hydroperoxide, suggesting that DirA has alkyl hydroperoxide reductase activity.

Arch Mal Coeur Vaiss, 1995 Jun, 88(6), 899 - 901
{Corynebacterium diphtheriae endocarditis complicated by septic arthritis and cerebral abscess}; Lehnert F et al.; The authors report a rare case of the mitis type Corynebacterium diphteriae endocarditis on a prosthetic valve complicated by septic arthritis and cerebral abscess . The authors underline the importance of regular transoesophageal echocardiographic control and underline the diagnostic value of ultrafast computed tomography for the diagnosis of aortic annular and interventricular septal abscesses in patients with mechanical prosthetic valves.

Thorax, 1995 Jun, 50(6), 661 - 7
Neutrophil sequestration in rat lungs; Brown GM et al.; BACKGROUND--The transit of neutrophils through the pulmonary microvasculature is prolonged compared with red blood cells and is increased further during cigarette smoking and in exacerbations of chronic obstructive pulmonary disease . The increased residence time (sequestration) of neutrophils in the pulmonary capillaries in these conditions may be the first step leading to the accumulation of cells within the lung interstitium and in the bronchoalveolar space, so potentiating lung damage . A rat model has been developed to investigate the factors which may influence neutrophil transit through the lung microvasculature . METHODS--Intratracheal instillation of the heat killed organism Corynebacterium parvum was used to induce an acute neutrophil alveolitis . Neutrophils and red blood cells were isolated from donor rats, labelled with two distinct radioisotopes, and then reinjected into recipient rats to assess their transit through the pulmonary circulation . To ascertain whether peripheral blood neutrophils were minimally altered by the isolation procedure their functional status in vitro was compared with that of inflammatory neutrophils in a number of assays commonly used as descriptors of neutrophil activation . The influence of neutrophil activation on the accumulation of cells in the lungs was assessed by comparing the lung sequestration of control neutrophils, isolated from peripheral blood, with that of inflammatory neutrophils obtained from bronchoalveolar lavage of inflamed rat lungs . Lung sequestration of neutrophils was defined as the fold increase in the ratio of neutrophils labelled with chromium-51 to red blood cells labelled with technetium-99m in lung tissue compared with the same ratio in peripheral blood . RESULTS--Sequestration of peripheral blood neutrophils occurred in control rat lungs as shown by a 17.5 (2.1) fold increase in the ratio of neutrophils to red blood cells in the pulmonary circulation compared with the ratio of these cells in the peripheral circulation . When inflammatory neutrophils, obtained by bronchoalveolar lavage from C parvum-treated animals, were injected into control rats, the increase was 90.6 (11.0) fold . Induction of an inflammatory response in the lung tissue of the recipient rat also caused an increase in the sequestration of control neutrophils compared with the same cells in control rat lungs which was, however, less marked than when inflammatory neutrophils were used (34.7 (4.7) fold) . The mean (SE) pressure developed on filtration of inflammatory neutrophils in vitro through a millipore filter (7.53 (0.2) cm H2O) was greater than that of peripheral blood neutrophils (1.18 (0.2) cm H2O) . Increased filtration pressure indicates a decrease in cell deformability and suggests that this may be a contributory factor to the increased sequestration of inflammatory neutrophils in the pulmonary vasculature . CONCLUSIONS--This study shows that there is sequestration of neutrophils in the pulmonary vasculature in normal rat lungs which increases in acute lung inflammation and when inflammatory neutrophils are injected into control animals . In this model changes in the neutrophil, such as cell deformability, may have a more important role in inducing increased neutrophil sequestration than the inflammatory response in the lungs.

J Am Podiatr Med Assoc, 1995 Jun, 85(6), 338 - 9
Osteomyelitis caused by Corynebacterium jeikeium; Boc SF et al.; A case study has been presented where C . jeikeium was isolated as the causative bacterium of an osteomyelitis of the fifth metatarsal . Partial amputation, local wound care, frequent and aggressive debridement, and appropriate antibiotics were all used with apparent success . The lack of complete patient follow-up prohibits the authors from declaring the infection cured; however, all signs of infection were absent immediately prior to discharge . The authors believe this to be the first reported case of Corynebacterium species as the bacterial isolate in confirmed osteomyelitis.

J Hosp Infect, 1995 Jun, 30 Suppl, 306 - 12
The return of Corynebacterium diphtheriae: the rise of non-toxigenic strains; Wilson AP; With the decline in incidence of diphtheria in Europe and the USA, many laboratories no longer routinely culture throat swabs for Corynebacterium diphtheriae . However, there is an outbreak of infection with toxigenic strains in Russia and most adults do not have protective levels of antibody . Non-toxigenic strains are known to cause local disease and lysogenic conversion probably occurs in vivo as well as in vitro . Non-toxigenic C . diphtheriae var . gravis, formerly quite rare, has been isolated with increasing frequency in the UK over the last five years . During prospective screening at one Sexually Transmitted Disease Clinic, six (1%) of 578 homosexual men were found to harbour the organism in the throat, four of them with clinical pharyngitis . Only one of 1696 heterosexual men and women were found to be carriers . Seven cases of endocarditis due to this organism were reported in a single year in Sydney, Australia and non-toxigenic C . diphtheriae var . mitis has caused four cases of endocarditis in Switzerland . Non-toxigenic strains are responsible for pharyngitis and occasional invasive disease and should be treated . Routine screening of throat swabs should not be abandoned.

J Vet Med Sci, 1995 Jun, 57(3), 515 - 7
Assignment of the bacterial agent of urinary calculus in young rats by the comparative sequence analysis of the 16S rRNA genes of corynebacteria; Takahashi T et al.; Comparative 16S rRNA gene sequencing was used to assign four isolates of spontaneous urinary calculus in young laboratory rats . The phylogenetic relationships among the rat isolates and selected species of corynebacteria were also inferred . Based on the homology and evolutionary distance analysis, the 16S rRNA genes of the rat isolates were almost identical with that of Corynebacterium renale ATCC 19412 . Also the results of the phylogenetic analysis showed a close relationship among the isolates and C . renale, but they were clearly different from C . pilosum, C . cystitidis, C . kutscheri and Rhodococcus equi . The results of the present study and previously published biochemical data demonstrate that the organism involving urinary infections in young rats is identified to be C . renale.

Biochem J, 1995 May 15, 308 ( Pt 1), 313 - 20
Prephenate dehydratase of the actinomycete Amycolatopsis methanolica: purification and characterization of wild-type and deregulated mutant proteins; Euverink GJ et al.; Prephenate dehydratase (PDT) is a key regulatory enzyme in L-phenylalanine biosynthesis in the Gram-positive bacterium Amycolatopsis methanolica . The PDT protein was purified to homogeneity (1957-fold) from wild-type cells with a final yield of 6.5% . It was characterized as a 150 kDa homotetrameric protein with a subunit size of 34 kDa . The first 35 N-terminal amino acids were identified, revealing highest similarity to the PDT proteins from Corynebacterium glutamicum and Bacillus subtilis . Kinetic studies showed that the A . methanolica PDT is allosterically inhibited by phenylalanine and activated by tyrosine . Phenylalanine caused an increase in the S0.5 for prephenate and a decrease in the Vmax . Tyrosine caused a decrease in the S0.5 for prephenate and an increase in the Vmax . Spontaneous o-fluoro- and p-fluoro-DL-phenylalanine-resistant mutants of A . methanolica were isolated . Kinetic studies with the partially purified PDT proteins of strains pFPhe32 and oFPhe84 showed that these mutant proteins had become (partly) insensitive to both phenylalanine inhibition and tyrosine activation.

Med Clin (Barc), 1995 May 13, 104(18), 699 - 701
{Corynebacterium pseudotuberculosis adenitis in a shepherd}; Bartolome J et al.; Corynebacterium pseudotuberculosis causes suppurative lymphadenitis in sheep and other domestic animals . Human infection has been reported in few instances . We report on a case of Corynebacterium pseudotuberculosis lymphadenitis in a human being . A 34-years old previously healthy shepherd was attended for presenting a painful lymph node in right groin with one year of evolution . Cultures of an aspirate and ganglionar tissue yielded growth of Corynebacterium pseudotuberculosis in pure culture . Histological examination of the excised lymph node showed a suppurative granulomatous lymphadenitis with areas of necrosis in which there were clusters of bacillary organisms . After surgical excision and administration of erythromycin clinical signs disappeared without complications . We have carried out a review of the literature and we have found no case reported in Spain . Corynebacterium pseudotuberculosis lymphadenitis in human beings is a rare entity that principally affects persons in contact with animals, principally sheep . Most cases have been reported in Australia . In accordance with the reviewed literature this is the first time this disease is reported in Spain . Corynebacterium pseudotuberculosis infection should be considered in the differential diagnosis of localized granulomatous lymphadenitis.

Cell Signal, 1995 May, 7(4), 313 - 8
Does nitric oxide play a role in liver function?
Milbourne EA, Bygrave FL.
Nitric oxide (NO) is becoming increasingly recognised as a signalling molecule in many organs, although its role in the liver remains to be fully elucidated . There is no doubt that liver cells can produce NO in response to a variety of stimuli including Corynebacterium parvum-infection, lipopolysaccharide (LPS) and a variety of cytokines . Within the liver, NO modulates some fundamental intracellular functions such as protein synthesis, mitochondrial electron transport and components of the citric acid cycle . Intercellular roles for NO in the liver may include drug metabolism and blood storage . Also, NO acts to protect the liver from immunological damage in models of hepatic inflammation . Understanding the role of NO in the liver may provide insight into the functioning of this organ in health and disease.

J Neurosci Res, 1995 May 1, 41(1), 8 - 14
Changes in neuronal mRNAs induced by a local inflammatory reaction; Lu X et al.; Injection of Corynebacterium parvum into the rat dorsal root ganglion has previously been shown to cause an inflammatory reaction dominated by macrophages and to enhance regeneration of the central axons of primary sensory neurons . Here, neuronal mRNAs that are modified by nerve transection were analyzed by in situ hybridization following injection of C . parvum into the dorsal root ganglion . Neuronal concentrations of mRNAs for the growth-associated protein (GAP-43) and the immediate early gene c-jun were increased by a local inflammatory response just as after axotomy . The concentration of mRNA for calcitonin gene-related peptide (CGRP) was also increased in a constant subpopulation of sensory neurons after injection of C . parvum in contrast to its decrease following axotomy . The results are consistent with the hypothesis that some of the responses to sensory neurons to axotomy are sustained by macrophages which accumulate within the dorsal root ganglion after nerve injury.

Mol Plant Microbe Interact, 1995 May-Jun, 8(3), 371 - 8
Characterization of a pathogen-induced potato catalase and its systemic expression upon nematode and bacterial infection; Niebel A et al.; We have isolated a cDNA encoding a catalase (Cat2St) by differential screening of a cDNA library constructed from potato roots infected with the cyst nematode Globodera pallida . Expression analysis confirmed the local induction of Cat2St and showed that it was highest at the adult stage of the parasite . It also revealed that Cat2St was induced in uninfected roots, stems, and leaves of infected plants . Localized and systemic induction of Cat2St was also observed upon root-knot nematode (Meloidogyne incognita) and root bacteria (Erwinia carotovora, Corynebacterium sepedonicum) infections . Based on sequence and expression analysis, Cat2St was found to belong to the recently described class II of dicotyledonous catalases, suggesting that these catalase isoforms could also be pathogen induced . Plant-parasitic nematodes are known to induce, in the roots of their hosts, highly metabolic feeding cells that function as nutritional sinks . Whereas the local induction of Cat2St is probably a consequence of an oxidative stress of metabolic nature, the systemic induction of Cat2St shows striking similarities with the induction of systemic acquired resistance (SAR) genes . The possible role of catalase in compatible plant-pathogen interactions is discussed.

J Ethnopharmacol, 1995 May, 46(2), 107 - 14
Regulation of hepatic macrophage function by oral administration of xiao-chai-hu-tang (sho-saiko-to, TJ-9) in rats; Fujiwara K et al.; The effect of Xiao-Chai-Hu-Tang (Sho-saiko-to, TJ-9), the extract of a mixture of 7 herbs, on hepatic macrophage function was studied using rats . Hepatic macrophages were activated by injection of Corynebacterium parvum or 70% partial hepatectomy . Oral administration of TJ-9 for 3 weeks did not affect the ability of these macrophages to produce superoxide anions evaluated in situ by liver perfusion with nitro blue tetrazolium (NBT) and phorbol myristate acetate (PMA) . However, the similar administration of TJ-9 attenuated the blocking of the activation after partial hepatectomy produced by pretreatment with gum arabic, a polysaccharide of high molecular weight . When gum arabic was added to the medium of rat hepatic macrophages cultured with normal rat sera, their ability to produce superoxide anions was reduced in a dose-related manner . This reduction was attenuated by changing the sera to the sera obtained from rats given oral doses of TJ-9 for 3 weeks . These results suggest that TJ-9 may improve the blocked function of hepatic macrophages in activation.

Int J STD AIDS, 1995 May-Jun, 6(3), 211 - 5
Pharyngeal flora in a sexually active population; Russell JM et al.; During a 7-week period 1141 patients attending the Genitourinary Clinic at Charing Cross Hospital completed a brief questionnaire and had pharyngeal swabs cultured for Neisseria spp, beta-haemolytic streptococci, corynebacterium and yeasts . The study included 397 heterosexual men, 492 heterosexual women, 189 homosexuals, 41 lesbians and 22 bisexual men and women . Four hundred and sixty patients (40%) admitted oro-genital contact in the preceding 2 weeks . The meningococcal carriage rate was 11.6% . Homosexuals had the highest carriage 23.8% and heterosexual females the lowest 5.9% . Significant differences in carriage rates were found between homosexual and heterosexual men (P < 0.0001), heterosexual men and women (P < 0.005) and between lesbian and heterosexual women (P < 0.025) . Recent oro-anal contact significantly increased meningococcal isolation (P < 0.001) . A significant association between beta-haemolytic streptococci and concomitant meningococcal carriage was also found (P < 0.01) . Sexual orientation and oro-genital contact influences both meningococcal and pharyngeal yeast isolation and should be considered when interpreting pharyngeal culture results.

Eur J Pediatr, 1995 May, 154(5), 381 - 3
Corynebacterium diphtheriae osteomyelitis in an immunocompetent child: a case report; Poilane I et al.; Septic osteomyelitis of the hip in a previously healthy child is described . A weakly toxigenic Corynebacterium diphtheriae strain was isolated from the bone aspirate . The results of the treatment were rapidly satisfactory, after surgical drainage and antibiotic therapy with pristinamycin . Conclusion: This case report shows that C . diphtheriae has not disappeared in the developed world and can be responsible of systemic infections.

Arch Oral Biol, 1995 May, 40(5), 385 - 91
Calcium and water diffusion in single-species model bacterial plaques; Rose RK et al.; The diffusion of tritiated water and 45Ca through single-species model plaques was measured in a diaphragm cell . Using plaques of Streptococcus downei, the apparent diffusion coefficient for tritiated water showed a small but significant decrease between pH 7.0 and 5.0 with a smaller, non-significant decrease for calcium . These effects are attributed to possible plaque shrinkage at reduced pH, which overcomes effects due to charge-dependent changes in permeability . Effective diffusion coefficients for 45Ca obtained from lag-time measurements were much reduced at low carrier concentrations, demonstrating a binding which correlated with previous results from equilibrium dialysis . Strep . sanguis and Corynebacterium matruchotii showed few differences from Strep . downei, although the corynebacterium previously demonstrated stronger calcium binding . Permeability to water and calcium were slightly affected by pH, while effective diffusion of calcium depended on concentration . This may have important implications for the process of mineral loss in dental caries.

J Clin Microbiol, 1995 May, 33(5), 1318 - 21
Comparison of E-test with broth microdilution and disk diffusion for susceptibility testing of coryneform bacteria; Martinez-Martinez L et al.; The susceptibilities of 135 coryneform bacteria isolated from clinical samples to ampicillin (AMP), cephalothin (CR), cefoxitin (FOX), cefotaxime (CTX), erythromycin (E), ciprofloxacin (CIP), tetracycline (TE), amikacin (AK), vancomycin (VA), and rifampin (R) were determined by disk diffusion, broth microdilution, and the E-test . The following species (number of isolates in parentheses) were included: Corynebacterium urealyticum (30), Corynebacterium minutissimum (20), coryneform CDC group ANF-1 (20), Corynebacterium striatum (20), Corynebacterium jeikeium (15), coryneform CDC group I2 (8), Listeria monocytogenes (7), Corynebacterium xerosis (5), and other coryneform bacteria (10) . Agreement within one twofold dilution between the E-test and broth microdilution was 31% (VA), 64% (AK), 71% (CTX), 77% (FOX and CIP), 79% (TE), 84% (AMP), 87% (E), and 88% (CR and R) . For the 1,350 combinations of microorganisms and antimicrobial agents, 85 (6.3%) discrepancies in interpretive category were found (4.2% minor, 1.2% major, and 0.9% very major) . Seventy (5.1%) disagreements in interpretive category were found between disk diffusion and the E-test (3.8% minor, 0.4% major, and 0.9% very major), and 85 (6.3%) disagreements were found between microdilution (reference method) and disk diffusion (4.2% minor, 0.5% major, and 1.5% very major) . MICs obtained with the E-test were highly reproducible . No category discrepancy was observed for VA, despite quantitative results . Considering interpretive categories, there is a good overall agreement between the three methods studied here, but further evaluation of current methodologies for susceptibility testing is required when considering coryneform bacteria and determination of quantitative activity of antimicrobial agents.

J Clin Microbiol, 1995 May, 33(5), 1080 - 3
Molecular epidemiology of Corynebacterium diphtheriae from northwestern Russia and surrounding countries studied by using ribotyping and pulsed-field gel electrophoresis; De Zoysa A et al.; A selection of 100 Corynebacterium diphtheriae isolates from asymptomatic carriers and clinical cases from five regions in northwestern Russia were examined . Six additional isolates from patients in Finland and Estonia with epidemiological links to Russia were also examined . All isolates were characterized by biotyping, toxigenicity testing, ribotyping, and pulsed-field gel electrophoresis (PFGE) . Hybridization of genomic DNA digested with BstEII revealed five ribotype patterns among the biotype gravis isolates (G1 through G5) and two patterns among the biotype mitis isolates (M1 and M2) . PFGE using SfiI was not able to distinguish between ribotypes G1, G2, and G4 . The predominant ribotype pattern, G1, found in cases of disease in all the areas studied, appears to be disseminating, in view of the isolates received from imported cases in Finland and Estonia . Among the 106 isolates examined, 68 produced pattern G1 and 24 produced pattern M1 . Most of the M1 isolates were from the Leningrad Oblast region . Distinct ribotypes such as G2, G3, G4, G5, and M2 could represent endemic disease.

Immunol Lett, 1995 May, 46(1-2), 101 - 6
Murine tumorlytic factor, immunologically distinct from tumor necrosis factor-alpha and -beta, induced in the serum of mice treated with a T-cell mitogen of Corynebacterium kutscheri; Kita E et al.; Murine tumorlytic factor (TF), immunologically distinct from murine tumor necrosis factor (TNF)-alpha and -beta, was purified to a homogeneity from the serum of mice injected with a T-cell mitogen of Corynebacterium kutscheri . The treated mouse serum was purified by Lentil lectin-Sepharose chromatography, DEAE-cellulose chromatography, preparative isoelectric focusing, and high-pressure liquid chromatography to the specific activity of 1.5 x 10(6) U/mg protein . TF was 42 kDa in its oligomeric form and 14 kDa in its monomeric form . TF activity was not impaired with hamster monoclonal antibody (mAb) to recombinant murine TNF-alpha and -beta and, reciprocally, rabbit antibody to TF neutralized the bioactivity of neither murine TNF-alpha nor -beta . TF was not precipitated with the mAb to murine TNF-alpha and -beta in Western blot analysis . The partial amino acid sequence of TF was at most 33% homologous to the 46-63 sequence of mouse TNF-beta . Thus, these results suggest that TF might be a novel tumorlytic factor which is immunologically distinct from mouse TNF-alpha and -beta.

Plasmid, 1995 May, 33(3), 168 - 79
The erythromycin resistance gene of the Corynebacterium xerosis R-plasmid pTP10 also carrying chloramphenicol, kanamycin, and tetracycline resistances is capable of transposition in Corynebacterium glutamicum; Tauch A et al.; The clinical isolate Corynebacterium xerosis M82B carries the 50-kb R-plasmid pTP10 that confers resistance to the antibiotics chloramphenicol, kanamycin, erythromycin, and tetracycline . A detailed restriction map of pTP10 was constructed by cloning and analyzing restriction fragments of pTP10 in Escherichia coli . The resistance determinants of pTP10 were located by studying the phenotype of the recombinant plasmids in E . coli and Corynebacterium glutamicum . Restriction patterns of fragments encoding the kanamycin and erythromycin resistances revealed striking similarity to the kanamycin resistance of transposon Tn903 and the erythromycin resistance on plasmid pNG2 from Corynebacterium diphtheriae, respectively . Expression of the resistance determinants in E . coli and C . glutamicum ATCC 13032 led to high resistance levels in both strains, with the exception of the tetracycline resistance gene, which could be expressed only in C . glutamicum . Furthermore, the erythromycin resistance gene was found to be located on a transposable element which is functional in C . glutamicum strains.

Microbiol Res, 1995 May, 150(2), 187 - 94
Insecticides: their effect on microorganisms and persistence in rice soil; Das AC et al.; A field experiment was conducted to investigate the effect of four insecticides, HCH, phorate, carbofuran and fenvalerate, at recommended doses on the preponderance of bacteria, actinomycetes and fungi . We also measured the persistence of the insecticides in the rhizosphere soil of rice . HCH and fenvalerate stimulated the proliferation of all of the microorganisms significantly . Phorate increased the population of bacteria and actinomycetes . Carbofuran accentuated the preponderance of actinomycetes in soil . Insecticides, in general, did not have marked influence on the proliferation of Bacillus, Streptomyces, Aspergillus and Fusarium in soil . However, we observed a stimulation of growth of Staphylococcus, Proteus and Sarcina with HCH, Pseudomonas, Corynebacterium, Erysipelothrix and Rhizopus with phorate, Serratia, Corynebacterium, Klebsiella, Escherichia, Rhizopus and Humicola with carbofuran, and Staphylococcus, Sarcina, Klebsiella and Nocardia with fenvalerate . On the other hand, there was an inhibition in growth of Pseudomonas, Micrococcus, Nocardia and Penicillium with HCH, of Pseudomonas, Micrococcus and Penicillium with carbofuran, and of Pseudomonas, Micrococcus and Micromonospora with fenvalerate . Different types of insecticides exhibited differential patterns of dissipation in soil . HCH had the highest persistence followed by phorate, carbofuran and fenvalerate, respectively.

Med Clin (Barc), 1995 Apr 22, 104(15), 561 - 4
{Corynebacterium urealyticum in kidney transplant patients}; Garcia Bravo M et al.; BACKGROUND: Corynebacterium urealyticum may produce severe urinary tract infections (UTI) in patients with renal transplantation (RT) . The aim of this study was to define the prevalence, clinical spectrum and risk factors for the development of symptomatic UTI in RT receptors with bacteriuria by C . urealyticum . METHODS: The clinical data of RT patients with bacteriuria by C . urealyticum diagnosed in the Hospital Doce de Octubre in Madrid from January 1990 to September 1993 were retrospectively reviewed . The patients corresponded to two clearly differentiated periods . In the first, the presence of C . urealyticum was not actively sought in the urine sample while in the second an intentional search was carried out in all the RT with a selective culture medium containing different antibiotics, Tween-80 and urea to facilitate C . urealyticum identification and growth . RESULTS: C . urealyticum was isolated in the urine of 46 patients (14% of the RT performed in the study period) . In the first phase 16 cases were diagnosed with 30 being found in the second with the selective medium . Bacteriuria by C . urealyticum was symptomatic in 18 patients (39%): 12 acute cystitis, one encrusted cystitis (IC), and 5 encrusted pyelitis (IP) . Of the symptomatic patients 39% had a history of prolonged vesical catheterization, 27% carried ureteral catheterization and 50% had undergone other urologic manipulations . The clinical consequences were important with development of obstructive uropathy and alteration in renal function (28%), need for surgery (33%) and graft loss (5.5%) . All the C . urealyticum strains were sensitive to vancomycin and teicoplanin which were useful in the treatment although the most severe cases (IC, IP) required surgery . CONCLUSIONS: The prevalence of UTI by Corynebacterium urealyticum is high in RT patients . Occasionally, these infections may have severe consequences, particularly in patients with a history of urologic manipulation, if early diagnosis is not performed and adequate antibiotic treatment given . A selective culture medium should be used to isolate C . urealyticum in RT patients.

Cell Immunol, 1995 Apr 15, 162(1), 1 - 7
Dibutyryl cyclic AMP protects Corynebacterium parvum-treated mice against lipopolysaccharide-induced lethal toxicity; Inoue H et al.; The effects of dibutyryl cyclic AMP (DBcAMP) on the lethal toxicity of lipopolysaccharide (LPS) were investigated in mice made hypersensitive to LPS by administration with Corynebacterium parvum (C . parvum) . The peritoneal macrophages in C . parvum-treated mice released a conspicuous level of TNF alpha in response to LPS in vitro . These macrophages exhibited much higher susceptibility to LPS-induced cytotoxicity than with resident or peptone-induced macrophages . Pretreatment of the C . parvum-induced macrophages with DBcAMP significantly inhibited the TNF alpha production in response to LPS and decreased the susceptibility to LPS-induced cytotoxicity in vitro . Similar to the findings seen in in vitro experiments, in vivo administration with DBcAMP significantly inhibited the TNF alpha release in the sera of C . parvum-treated mice after LPS challenge and consequently decreased the susceptibility to LPS-induced shock . These results suggest that raising level of intracellular cAMP protects C . parvum-treated mice from hypersensitivity to lethal toxicity of LPS through downregulating TNF alpha synthesis.

Gene, 1995 Apr 14, 156(1), 113 - 8
Toxic phospholipases D of Corynebacterium pseudotuberculosis, C . ulcerans and Arcanobacterium haemolyticum: cloning and sequence homology; McNamara PJ et al.; The genes encoding toxic phospholipases D (PLD) from Corynebacterium pseudotuberculosis (Cp)biovar equi and C . ulcerans (Cu) have been cloned and sequenced . The deduced proteins are 307 amino acids (aa) in length and include a putative signal sequences of 26-aa . A molecular mass of 31.2 and 31.0 kDa and pI values of 8.84 and 6.73 are predicted for the secreted (mature) proteins from Cp and Cu, respectively . Comparison of the deduced primary structure of the two proteins to those of the PLD produced by Cp biovar ovis and Arcanobacterium haemolyticum (Ah) revealed that the four enzymes share 64-97% identity . The aa sequences of this group of proteins were unique when compared to the sequences of other phospholipases in GenBank and were found to share only small regions of homology with other proteins, including two conserved domains of glyceraldehyde-3-phosphate dehydrogenase (G3PD) . The similarity of PLD from Cp biovar equi, Cu and Ah to the PLD of Cp biovar ovis suggests that these enzymes may act as virulence determinants.

Biokhimiia, 1995 Apr, 60(4), 644 - 51
{Stability of a new product of oxidative stress in bacterial cells}; Demina GR et al.; Data on 32P-label incorporation with subsequent addition of non-radiolabelled o-phosphate suggest that the new phosphorus compound, 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEC), accumulated in the cells of some bacterial species in response to oxidative stress does not rapidly exchange phosphorus with external o-phosphate 3 hours after the introduction of its synthesis inducers into the Corynebacterium ammoniagenes culture . The accumulated MEC is retained in the cells despite the action of the cell wall synthesis inhibitor, chloramphenicol, or the energetic poisons, KCN and iodoacetate and also under anaerobic conditions . It has been shown that incubation of the cell-free lysate of a non-induced culture, Micrococcus luteus, with MEC does not result in MEC hydrolysis; therefore, MEC accumulation after the redox-mediator addition is hardly due to the hydrolase inactivation but, rather, is due to the activation of the MEC-synthesizing enzyme . The cells of C . ammoniagenes incorporate 32P from {32P}MEC but not 14C from {14C}MEC . This points to MEC hydrolysis prior to the uptake of its phosphoryl fragment by the cells . In this case 32P is found in the fractions differing by their position from MEC fractions . Experiments with sheep erythrocytes and mouse splenocytes revealed that MEC (10-100 micrograms per 1,000,000 splenocytes) does not influence the antibody production by these cells, whereas used at concentrations of 200-550 micrograms per 1,000,000 cells, MEC enhances the antibody production . However, while doing so, MEC causes the destruction of a considerable portion of splenocytes and sheep erythrocytes.

Biosci Biotechnol Biochem, 1995 Apr, 59(4), 694 - 702
Nucleotide sequence of the FAD synthetase gene from Corynebacterium ammoniagenes and its expression in Escherichia coli; Nakagawa S et al.; The nucleotides of a bifunctional enzyme FAD synthetase gene, which showed both flavokinase and ATP:FMN adenylyltransferase activities, from Corynebacterium ammoniagenes were sequenced . The FAD synthetase gene product consisted of 338 amino acids and had a calculated molecular weight of 37,712 . The deduced protein sequence of the FAD synthetase shared a homology with those of the protein X of Escherichia coli, which has been reported to have both flavokinase and ATP:FMN adenylyltransferase activities like the FAD synthetase of C . ammoniagenes, and the protein X of Pseudomonas fluorescens . From the analysis of the flanking sequences of the FAD synthetase gene, the gene organization and the operon structure around the FAD synthetase gene of C . ammoniagenes were thought to be different from those of Gram-negative bacteria . An over-expression system of the FAD synthetase of C . ammoniagenes was constructed in E . coli to study the structure and function of the protein . Under the tandem tryptophan promoter, the FAD synthetase activity increased 2231 times compared to that of non-transformed C . ammoniagenes.

Appl Microbiol Biotechnol, 1995 Apr, 43(1), 83 - 8
High-yield production of diphtheria toxin mutants by high-density culture of C7 (beta)tox+ strains grown in a non-deferrated medium; Fass R et al.; A high-density growth approach was utilized to produce mutated diphtheria toxin from two strains of Corynebacterium diphtheria: C7 (beta)(tox-201,tox-9) and C7 (beta)(tox-107) . The cross-reacting mutants (CRM) of the diphtheria toxin are CRM9 and CRM107; both of them carry the mutation in their binding site and, as a result, have 1/300 of the systemic toxicity of the wild-type diphtheria toxin . Since iron inhibits diphtheria toxin production, the traditional approach has been to grow the bacteria in a very low iron concentration . The procedure described here involved the use of a modified, non-deferrated, growth medium that provided fast and high-density growth of the bacteria, and which, when associated with simultaneous depletion of glucose and iron, enhanced the toxin production . Oxygen-enriched air was supplied to enable the bacteria to grow to a cell density giving an absorbance of 70 at 600 nm (15-20 g/l dry weight) . The maximum toxin concentration in the culture supernatant was 150 mg/l . The CRM products, which remained stable following microfiltration and ultrafiltration, could be easily purified using a two-step chromatography procedure.

Appl Microbiol Biotechnol, 1995 Apr, 43(1), 76 - 82
Effect of different levels of aspartokinase on the lysine production by Corynebacterium lactofermentum; Jetten MS et al.; A 2.9-kb SacI fragment containing the ask-asd operon, encoding aspartokinase and aspartatesemialdehyde dehydrogenase, was cloned from an aminoethylcysteine-resistant, lysine-producing Corynebacterium lactofermentum strain . Enzymatic analysis showed that the aspartokinase (ASK) activity was completely resistant to inhibition by mixtures of lysine and threonine . Comparison of the deduced amino acid sequence of the beta submit of the ask gene showed three amino acid residue changes with ask gene encoding wild-type, feedback-sensitive enzymes . Three C . lactofermentum strains, one being aspartokinase-negative, one carrying two ask genes on the chromosome and one having a sixfold higher specific ASK activity than the parental strain, were constructed by transconjugation and electroporation, and used to analyse the role of ASK in the lysine production by C . lactofermentum . The results indicate that, in this study, feed-back-resistant ASK is necessary for high-level lysine production, but dispensable for lysine and diaminopimelate synthesis required for cell growth.

FEMS Microbiol Lett, 1995 Apr 1, 127(3), 263 - 6
Functional expression of the glutamate uptake system from Corynebacterium glutamicum in Escherichia coli; Burkovski A et al.; Glutamate uptake in the Gram-positive Corynebacterium glutamicum is mediated via a binding protein-dependent transport system, which is encoded by the gluABCD gene cluster . Cloning of these genes in an expression vector and subsequent transformation of the resulting plasmid allows different strains of the Gram-negative bacterium Escherichia coli to grow on glutamate as sole carbon and nitrogen source . However, overexpression of the glutamate uptake system results in growth inhibitory effects, probably due to the particular topology of the binding protein.

Appl Environ Microbiol, 1995 Apr, 61(4), 1610 - 3
Expression and secretion of heterologous proteases by Corynebacterium glutamicum; Billman-Jacobe H et al.; Genes encoding the basic protease of Dichelobacter nodosus (bprV) and the subtilisin of Bacillus subtilis (aprE) were cloned and expressed in Corynebacterium glutamicum . In each case, enzymatically active protein was detected in the supernatants of liquid cultures . While the secretion of subtilisin was directed by its own signal peptide, the natural signal peptide of the bprV basic protease did not facilitate secretion . A hybrid aprE-bprV gene in which the promoter and signal peptide coding sequences of subtilisin replaced those of bprV could be expressed, and basic protease was secreted by C . glutamicum . Expression of these proteases in C . glutamicum provides an opportunity to compare protein secretion from this gram-positive host with that from other gram-positive and gram-negative bacteria.

Br J Ophthalmol, 1995 Apr, 79(4), 347 - 9
Adherence of bacteria to intraocular lenses: a prospective study; Doyle A et al.; AIMS--The study was designed to investigate the bacterial flora of the operating field during routine cataract surgery and the source of intraocular lens contamination during the surgery . METHODS--The normal flora of the external eye and fornices of 17 patients undergoing selective cataract surgery was determined preoperatively . Swabs taken from the eyelid surface and lashes showed coagulase negative staphylococci (CNS) in 90%, Propionibacterium acnes in 62%, Corynebacterium sp in 18%, and Peptostreptococcus in 3% of the patients . The lower fornices of 70% had CNS, 47% P acnes, 6% Staphylococcus aureus, 6% Corynebacterium sp, and 6% Candida . RESULTS--A sterile PMMA intraocular lens was touched on the upper bulbar conjunctiva immediately before the surgery . Eighty two per cent of lenses grew CNS, 18% P acnes, 18% Bacillus sp, 12% S aureus, and 6% Corynebacterium sp . A second sterile PMMA intraocular lens was left on the drape and near the eye during surgery . Forty seven per cent of these cultured CNS, 12% Corynebacterium sp, and 6% Bacillus sp . A high count of bacteria in the operating field, especially CNS and P acnes can contribute to postoperative inflammation and endophthalmitis . CONCLUSION--Special measures are needed before and during the surgery to reduce the chance of intraocular inoculation of these bacteria . Use of proper culture media and techniques are necessary to identify these organisms, especially anaerobes, in postoperative inflammation.

Eur Respir J, 1995 Apr, 8(4), 651 - 3
Pulmonary nodules due to Corynebacterium ulcerans; Dessau RB et al.; A 53 year old male with symptoms of coughing for 6 months presented with bilateral multiple pulmonary nodules suggestive of metastatic disease . By surgical resection 4 out of 6 nodules were removed . Histopathological examination showed granulomatous necrotizing inflammation with growth of Corynebacterium ulcerans, which did not produce diphtheria toxin . The patient was treated with penicillin for 1 week . Follow-up for 2 yrs showed no sign of recurrence.

Int J Syst Bacteriol, 1995 Apr, 45(2), 240 - 5
A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences; Chun J et al.; Partial sequences of the 16S rRNA genes of the type strains of nine species of the genus Nocardia were determined following the isolation and cloning of the amplified genes . These sequences were aligned with the sequences of representatives of the genera Corynebacterium, Gordona, Mycobacterium, Rhodococcus, and Tsukamurella, and phylogenetic trees were inferred by using the Fitch-Margoliash and neighbor-joining methods . The genus Nocardia formed a distinct clade that was most closely associated with the genus Rhodococcus . The average level of sequence similarity found among the type strains of the Nocardia species was 97.2 +/- 0.7% . Two sublines were recognized within the Nocardia clade; one encompassed Nocardia asteroides and related species, and the other encompassed Nocardia otitidiscaviarum and allied taxa . Separation of the two sublines is based on differences in helix 37-1 . The results of isoprenoid quinone analyses provided evidence that nocardiae can be distinguished from all other actinomycete taxa on the basis of their characteristic menaquinone profiles . Nocardiae typically contain hexahydrogenated menaquinones with eight isoprene units in which the two end units are cyclized.

JAMA, 1995 Mar 15, 273(11), 849 - 53
Interchangeability of conjugated Haemophilus influenzae type b vaccines in infants; Anderson EL et al.; OBJECTIVE--To evaluate the safety and immunogenicity of two Haemophilus influenzae type b (Hib) conjugate vaccines when administered in serial combination . These vaccines consisted of Hib capsular polysaccharide polyribosyl-ribitol phosphate (PRP) conjugated to the meningococcal outer membrane protein (OMP) complex (PRP-OMP) and H influenzae oligosaccharide conjugated to a mutant toxin (CRM197) isolated from Corynebacterium diphtheriae (HbOC) . DESIGN--Randomized, double-blind, clinical trial evaluating five Hib vaccination regimens . SETTING--Vaccine Treatment and Evaluation Units and affiliated private pediatric practices at Saint Louis (Mo) University, Vanderbilt University, Nashville, Tenn, and Baylor College of Medicine, Houston, Tex . PATIENTS--A total of 497 healthy 2-month-old infants scheduled to receive routine immunization . INTERVENTION--Participants received either PRP-OMP or HbOC given as recommended by the manufacturer, PRP-OMP at 2 and 6 months, HbOC at 2 months, then PRP-OMP at 4 and 6 months, or PRP-OMP at 2 months and then HbOC at 4 and 6 months . Unconjugated PRP was given at 15 months to evaluate priming . RESULTS--Geometric mean antibody concentrations differed significantly among the groups following the second and third immunizations of the primary series and following booster immunization with unconjugated PRP . On each occasion, the groups receiving serial combinations of PRP-OMP and HbOC achieved mean antibody concentrations that equalled or exceeded those of the groups receiving a single product . Adverse reactions did not vary by group . CONCLUSIONS--The studied sequential combinations of Hib vaccines were safe and at least as immunogenic as either vaccine alone.

J Bacteriol, 1995 Mar, 177(5), 1152 - 8
Structure of the gluABCD cluster encoding the glutamate uptake system of Corynebacterium glutamicum; Kronemeyer W et al.; To assess the mechanism and function of the glutamate uptake system of gram-positive Corynebacterium glutamicum, a mutant deficient in glutamate uptake was isolated and was then used to isolate a DNA fragment restoring this deficiency . In a low-copy-number vector, this fragment resulted in an increased glutamate uptake rate of 4.9 nmol/min/mg (wild type, 1.5 nmol/min/mg) . In addition, carbon source-dependent regulation of the glutamate uptake system was determined with the fragment, showing that the entire structures required for expression and control reside on the fragment isolated . Sequencing of 3,977 bp revealed the presence of a four-gene cluster (gluABCD) with deduced polypeptide sequences characteristic of a nucleotide-binding protein (GluA), a periplasmic binding protein (GluB), and integral membrane proteins (GluC and GluD), identifying the glutamate transporter as a binding protein-dependent system (ABC transporter) . This identification was confirmed by the kinetic characteristics obtained for cells grown in the presence of globomycin, which exhibited an increased Km of 1,400 microM (without globomycin, the Km was 1.5 microM) but a nearly unaltered maximum velocity . By applying gene-directed mutagenesis, a strain with the entire cluster deleted was constructed . With this mutant, the glutamate uptake rate was reduced from 1.4 to less than 0.1 nmol/min/mg, which is proof that this system is the only relevant one for glutamate uptake . With this strain, the glutamate excretion rate was unaffected (18 nmol/min/mg), showing that no component of gluABCD is involved in export but rather that a specific machinery functions for the latter purpose.

Ann Emerg Med, 1995 Mar, 25(3), 390 - 403
Group A streptococcal tonsillopharyngitis: cost-effective diagnosis and treatment; Pichichero ME; Most patients who seek medical attention for sore throat are concerned about streptococcal tonsillopharyngitis, but fewer than 10% of adults and 30% of children actually have a streptococcal infection . Group A beta-hemolytic streptococci (GAS) are most often responsible for bacterial tonsillopharyngitis, although Neisseria gonorrhea, Arcanobacterium haemolyticum (formerly Corynebacterium haemolyticum), Chlamydia pneumoniae (TWAR agent), and Mycoplasma pneumoniae have also been suggested as possible, infrequent, sporadic pathogens . Viruses or idiopathic causes account for the remainder of sore throat complaints . Reliance on clinical impression to diagnose GAS tonsillopharyngitis is problematic; an overestimation of 80% to 95% by experienced clinicians typically occurs for adult patients . Overtreatment promotes bacterial resistance, disturbs natural microbial ecology, and may produce unnecessary side effects . Existing data suggest that rapid GAS antigen testing as an aid to clinical diagnosis can be very useful . When used appropriately, it is sensitive (79% to 88%) in detecting GAS-infected patients and is specific (90% to 96%) and cost-effective . Penicillin has been the treatment of choice for GAS tonsillopharyngitis since the 1950s; 10 days of treatment are necessary for bacterial eradication . A single IM injection of benzathine penicillin is effective and obviates compliance issues . Until the early 1970s, the bacteriologic failure rate for the treatment of GAS tonsillopharyngitis ranged from 2% to 10% and was attributed to chronic GAS carriers . Since the late 1970s, the penicillin failure rate has frequently exceeded 20% in published reports . Explanations for recurrent GAS tonsillopharyngitis include poor patient compliance; reacquisition from a family member or peer, copathogenic colonization by Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, anaerobes that inactivate penicillin with beta-lactamase, or all these organisms; suppression of natural immune response by too-early administration of antibiotics; GAS tolerance to penicillin; antibiotic eradication of normal pharyngeal flora that normally act as natural host defenses; and establishment of a true carrier state . When therapy fails, milder symptoms may occur during the relapse . Several antimicrobials have demonstrated superior efficacy compared with penicillin in eradicating GAS and are administered less frequently to enhance patient compliance . In previously untreated GAS throat infections, cephalosporins produce a 5% to 22% higher bacteriologic cure rate; after a penicillin treatment failure, these differences are greater . Amoxicillin/clavulanate and the extended-spectrum macrolides clarithromycin and azithromycin may also produce enhanced bacteriologic eradication in comparison to penicillin.(ABSTRACT TRUNCATED AT 400 WORDS)

J Clin Microbiol, 1995 Mar, 33(3), 759 - 61
Prosthetic valve endocarditis caused by Corynebacterium afermentans subsp . lipophilum (CDC coryneform group ANF-1); Sewell DL et al.; Corynebacteria are important causes of endocarditis in individuals with valvular prostheses . We report the first published case of prosthetic valve endocarditis caused by the newly defined species Corynebacterium afermentans subsp . lipophilum (former CDC coryneform group ANF-1) . The isolate was recovered from a perivalvular abscess specimen and 5 of 15 Bactec blood cultures after 7 to 15 days of incubation . The isolation, identification, and susceptibility testing of Corynebacterium species are discussed.

FEMS Microbiol Lett, 1995 Mar 1, 126(3), 271 - 6
Taxonomy of Corynebacterium diphtheriae and related taxa, with recognition of Corynebacterium ulcerans sp . nov . nom . rev; Riegel P et al.; Levels of genomic DNA relatedness were determined using a S1 nuclease procedure for reference bacteria identified as biotypes of Corynebacterium diphtheriae, biovars of Corynebacterium pseudotuberculosis, and 'Corynebacterium ulcerans' . These results showed that the three species are separate taxa at the genomospecies level whereas biotypes and biovars are closely related genomically within each species . Phylogenetic analyses of small-subunit rDNA sequences revealed that 'Corynebacterium ulcerans' forms a tight cluster with Corynebacterium pseudotuberculosis within the robust branch that groups all Corynebacterium sequenced to date . Therefore, we propose that the species incertae sedis 'C . ulcerans' should be conclusively recognized as a distinct species within the genus Corynebacterium with strain CCUG 2708 = NCTC 7910 as type strain . This species is characterized by urease production and fermentation of glycogen.

Vet Immunol Immunopathol, 1995 Mar, 45(1-2), 127 - 37
Effective immune protection of pigs against cysticercosis; Nascimento E et al.; A scolex protein antigen (SPA) was prepared from cysticerci of Taenia solium obtained from naturally infected pigs . Yorkshire pigs were vaccinated with SPA plus incomplete Freund's adjuvant (IFA) or with SPA plus Corynebacterium parvum (CP) . Controls were given IFA plus phosphate-buffered saline (PBS) or CP plus PBS . All animals were given three subcutaneous injections at 20-day intervals . Ten days after the third injection, the pigs were fed with 10(4) viable eggs of T . solium . All pigs developed a delayed type hypersensitivity, and a transient eosinophilia after the first dose of vaccine . High titers of specific antibodies were detected in the sera of vaccinated animals and in infected controls . A protection level of 71.43% was recorded in animals vaccinated with SPA plus IFA and of 75.00% in those vaccinated with SPA plus CP.

Int Immunol, 1995 Mar, 7(3), 481 - 91
Lymphocytes infiltrating the CNS during inflammation display a distinctive phenotype and bind to VCAM-1 but not to MAdCAM-1; Engelhardt B et al.; The nature of inflammatory lymphocytes recruited to the CNS has been studied in a model of chronic inflammation . Injection of killed Corynebacterium parvum into the cortex of the mouse brain produces a circumscribed inflammatory cellular infiltrate around the injection site, and recruited mononuclear inflammatory cells (IC) can be isolated for flow cytometric analysis . The majority of IC were T cells . In comparison with the predominant naive population of mesenteric lymph node T cells, IC T cells express much higher levels of CD44, LFA-1 and ICAM-1, and lower levels of CD45RB, features commonly associated with memory (previously activated) cells . In addition, in contrast to the L-selectin+ alpha 6-integrinlow phenotype of naive lymph node T cells, IC T cells lacked L-selectin and were alpha 6-integrin- . Mac-1, recently proposed as another marker of memory T cell differentiation, was not displayed by IC T cells, suggesting that Mac-1 expression may be heterogeneous among memory T cell subsets . A subset of mesenteric lymph node (MLN) T cells, probably representing activated T cells undergoing the naive to memory transition, but not of IC T cells, expressed high levels of alpha 6-, beta 7- and alpha E-integrin . IC and MLN naive T cells expressed comparable levels of alpha 4-integrin, but IC T cells stain poorly with anti-beta 7 mAbs and with mAb DATK 32, specific for the alpha 4 beta 7 heterodimeric lymphocyte homing receptor for the mucosal addressin MAdCAM-1, suggesting that these inflammatory cells express more alpha 4 beta 1 than alpha 4 beta 7 . Consistent with this, in in vitro adhesion assays, brain IC bound better than MLN cells to the alpha 4 beta 1 integrin ligand VCAM-1 and the LFA-1 ligand ICAM-1 but adhered very poorly to the alpha 4 beta 7 ligand MAdCAM-1 . These findings are consistent with and extend previous immunohistological studies of T cells in murine experimental autoimmune encephalomyelitis, and demonstrate a distinctive phenotype for lymphocytes being present in the chronically inflamed brain.

Urology, 1995 Feb, 45(2), 223 - 9
A prospective study of microorganisms in urine and bladder biopsies from interstitial cystitis patients and controls; Keay S et al.; OBJECTIVES . Interstitial cystitis (IC) is a chronic inflammatory condition of the bladder of unknown etiology . We tested the hypothesis that a microorganism would be found at higher prevalence in urine or bladder tissue from women with IC than from control women . METHODS . Urine and bladder tissue were obtained at cystoscopy from 11 IC patients and 7 control subjects . These specimens were cultured for a variety of fastidious and nonfastidious bacteria, mycobacteria, fungi, and viruses . In addition, special staining techniques were used to examine biopsy specimens and cytospun urine, and tissue sections and outgrowths of explanted bladder cells were examined by electron microscopy . RESULTS . Cultures of urine from 6 of 11 IC patients grew five different bacteria (Corynebacterium sp . Klebsiella pneumoniae, Lactobacillus sp, Streptococcus constellatus, and Streptococcus morbillorum), human cytomegalovirus, or Torulopsis glabrata; one of these organisms (Lactobacillus sp) was found in urine from 2 patients . Although contamination by urethral organisms is possible, the prevalence of microorganisms in urine of IC patients (6 of 11) was significantly greater than in urine of control subjects (0 of 7) (P < 0.05) . Acridine orange staining revealed rods with appropriate morphology in urine from 4 of the 5 IC patients who had positive bacterial cultures and yeastlike organisms in urine and bladder tissue specimens that grew Torulopsis . Additionally, rodlike organisms were seen in urine from 2 IC patients with negative bacterial cultures and cocci were seen in the urine of 1 control patient . Biopsy specimens from 2 IC patients grew Torulopsis sp or Lactobacillus sp, in agreement with the results of acridine orange staining and culture of urine from these patients; in contrast, specimens from 3 control subjects grew small numbers of Pseudomonas sp or Staphylococcus epidermidis, but no organisms were cultured from urine or seen in acridine orange-stained tissue smears . All other cultures and stains were negative . CONCLUSIONS . These data do not provide evidence that IC is associated with infection or colonization by a single microorganism . However, they do generate the hypothesis that the prevalence of microorganisms, especially bacteria at low concentrations, is greater in the urine of IC patients than of control subjects . If these results are confirmed by other controlled studies, the question of whether the presence of these organisms is a cause or a result of IC should be addressed.

J Bacteriol, 1995 Feb, 177(3), 774 - 82
Cloning, sequence analysis, expression, and inactivation of the Corynebacterium glutamicum icd gene encoding isocitrate dehydrogenase and biochemical characterization of the enzyme; Eikmanns BJ et al.; NADP(+)-dependent isocitrate dehydrogenase (ICD) is an important enzyme of the intermediary metabolism, as it controls the carbon flux within the citric acid cycle and supplies the cell with 2-oxoglutarate and NADPH for biosynthetic purposes . In the amino acid-producing organism Corynebacterium glutamicum, the specific activity of ICD was independent of the growth substrate and of the growth phase at approximately 1 U/mg, indicating that this enzyme is constitutively formed . The ICD gene, icd, was isolated, subcloned on a plasmid, and introduced into C . glutamicum . Compared with the wild type, the recombinant strains showed up to 10-fold-higher specific ICD activities . The nucleotide sequence of a 3,595-bp DNA fragment containing the icd gene was determined . The predicted gene product of icd consists of 739 amino acids (M(r) = 80.091) and showed 58.5% identity with the monomeric ICD isozyme II from Vibrio sp . strain ABE-1 but no similarity to any known ICD of the dimeric type . Inactivation of the chromosomal icd gene led to glutamate auxotrophy and to the absence of any detectable ICD activity, suggesting that only a single ICD is present in C . glutamicum . From an icd-overexpressing C . glutamicum strain, ICD was purified and biochemically characterized . The native ICD was found to be a monomer; to be specific for NADP+; to be weakly inhibited by oxaloacetate, 2-oxoglutarate, and citrate; and to be severely inhibited by oxaloacetate plus glyoxylate . The data indicate that ICD from C . glutamicum is structurally similar to ICDs from bacteria of the genera Vibrio, Rhodomicrobium, and Azotobacter but different from all other known procaryotic and eucaryotic ICDs.

Am J Ophthalmol, 1995 Feb, 119(2), 181 - 8
Microorganisms cultured from the anterior chamber of ruptured globes at the time of repair; Ariyasu RG et al.; PURPOSE: We studied events leading to the development of posttraumatic endophthalmitis by examining the significance of 15 factors on microbial contamination of injured eyes . METHODS: A prospective study was done of 30 ruptured globes in patients admitted to an urban medical center . Cultures were taken from the conjunctiva before and after preoperative disinfection and from the anterior chamber at the beginning and end of wound repair . Twenty-five of 30 patients received a three-day regimen of intravenous antibiotics that were begun before surgery . RESULTS: Anterior chamber samples grew microorganisms in ten (33%) of 30 eyes, with positive cultures recovered from specimens taken at the beginning of wound repair in eight eyes and at the end of wound repair in six eyes . Contamination with indigenous flora may have occurred at the time of injury in one eye and during repair in another eye . Microbes recovered included Staphylococcus, Corynebacterium, and Aspergillus species . No patient developed endophthalmitis . Of the 15 factors studied, only intravenous antibiotics significantly decreased the incidence of positive anterior chamber cultures in eyes treated before wound repair compared with eyes not receiving such therapy (P = .002) . CONCLUSIONS: Despite the frequency of anterior chamber microbial contamination during injury or repair of the wound, with our treatment protocol and the presence of physiologic mechanisms to reduce intraocular microbes, no eyes developed clinical endophthalmitis . With our limited sample size only intravenous antibiotic therapy was found significantly to reduce anterior chamber microorganisms at the time of surgical repair, supporting their prophylactic use against the development of posttraumatic endophthalmitis.

Lab Anim Sci, 1995 Feb, 45(1), 6 - 10
Natural habitats of Corynebacterium kutscheri in subclinically infected ICGN and DBA/2 strains of mice; Amao H et al.; Subclinically infected mice of ICGN and DBA/2 strains housed in a conventionally managed colony were examined to determine natural habitats of Corynebacterium kutscheri . At 5, 7, 9, 12 and 13 months after initial isolation of the organism from oral cavity and cecal contents of five ICGN mice, attempts were made to isolate C . kutscheri from 19 sites using a new selective medium, furazolidone-nalidixic acid-colimycin agar . From the initial survey to 13 months, C . kutscheri was isolated from 27 of 29 ICGN mice (93.1%) and 9 of 10 DBA/2 mice (90%) . In contrast, antibody against C . kutscheri was detected in only 3 of 29 ICGN mice (10.3%) . None of the mice manifested distinct clinical signs of infection, and only 1 ICGN mouse had macroscopic lesions such as hepatic abscess and large spleen . In 21 ICGN and 9 DBA/2 mice that harbored the organism without macroscopic lesions, the organisms were most frequently isolated from the oral cavity (ICGN:100%, DBA/2:66.7%), cecum (ICGN:95.2%, DBA/2:100%), and colon and rectum (ICGN:95.2%, DBA/2:100%) . Remarkable differences between the two mouse strains were observed in colonization of the nasal cavity (ICGN:85.7%, DBA/2:0%) and trachea (ICGN:71.4%, DBA/2:33.3%) . In mice of both strains, the organisms rarely colonized the lung, liver, and kidney . Mean numbers of organisms in the cecum, and colon and rectum ranged from 10(4.1) to 10(4.6) colony-forming units/g and were significantly higher in comparison with those in the small intestine (P < 0.01, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Lab Anim Sci, 1995 Feb, 45(1), 11 - 4
Natural and subclinical Corynebacterium kutscheri infection in rats; Amao H et al.; Distribution of Corynebacterium kutscheri was determined in 41 rats housed in a conventionally managed colony that were infected naturally and subclinically . At 2, 5, 10, 20 and 25 months after initial isolation of C . kutscheri, attempts were made to isolate C . kutscheri from 17 sites, with a new selective medium, FNC agar . In total, the prevalence (97.6%) of C . kutscheri isolation was significantly (P < 0.001) higher than the frequency (70.7%) of antibody detection . None of the rats manifested any distinct clinical signs of disease and macroscopic lesions caused by C . kutscheri were not detected . In 40 rats with subclinical infection, the organisms were most frequently isolated from the oral cavity, esophagus, cecal contents, and colon and rectum (> 95.0%) . The isolation rate was next highest in the trachea, submaxillary lymph nodes, and nasal cavity (47.5 to 52.5%) . The organisms hardly colonized the lung, liver, and kidney . Mean numbers of organisms found in the esophagus, cecal contents, and colon and rectum ranged from 10(3.9) to 10(4.2) CFU/g, and were significantly (P < 0.05, P < 0.01) high in comparison with those in the lung . These results indicated that many healthy rats in the naturally infected colony harbored C . kutscheri, and the organisms colonized the oral cavity, esophagus, cecal contents, and colon and rectum most frequently.

Isr J Med Sci, 1995 Feb-Mar, 31(2-3), 160 - 2
Management of malignant pleural effusion secondary to breast cancer: talc pleurodesis and pleuroperitoneal shunting; Saute M; Malignant pleural effusion is a frequent complication of metastatic breast cancer, leading to a significant degree of morbidity . Drainage of the effusion by thoracocentesis and subsequent pleurodesis is an established means of symptomatic relief in these patients . Several sclerosing agents have been reported in the literature, including doxycylin, minocyclin, tetracyclines, bleomycin, cisplatin, etoposide, fluorouracil, interferon-beta, Corynebacterium parvum, and talc which gives the best results . The condition of the lung's parenchyma must be evaluated prior to the procedure to rule out lymphangitis carcinomatosa or bronchial obstruction that would impair the expansion of the lung . In these situations, the implantation of a pleuroperitoneal shunt is an alternative to be considered.

Int J Parasitol, 1995 Feb, 25(2), 229 - 39
Corneal virulence, cytopathic effect on human keratocytes and genetic characterization of Acanthamoeba; Badenoch PR et al.; Acanthamoeba keratitis is a sight-threatening complication of corneal trauma or contact lens wear . Although the majority of corneal isolates of Acanthamoeba belong to Group II in the Pussard-Pons classification based on cyst morphology, they have been placed in at least six species and their genetic relatedness is uncertain . The aim of this study was to determine the virulence of, and the relationship among, strains derived from the cornea, the nasal mucosa, and other environmental sources . To assess virulence, 10(4) trophozoites of each strain were incubated with monolayers of human corneal fibroblasts . By day 7, 12 of 29 strains tested had induced significant cytopathic changes . In addition, inocula of 10(4) cysts or trophozoites with 10(6) Corynebacterium xerosis were injected into the corneas of Porton rats; 11 amoebic strains induced infection within 7 days . The correlation between the virulence of trophozoites in vitro and in vivo was 86% . Using allozyme electrophoresis, 23 Acanthamoeba strains clustered into 5 major phylogenic divisions . Three divisions contained one or more strains that were virulent in the rat cornea . Virulent Pussard-Pons Group II strains clustered tightly to a fixed allelic difference of 13.6% . The eight corneal isolates clustered to 33%, dividing into three lineages . Five avirulent nasal isolates were strongly differentiated from other Group II strains . The results were not in accord with species designations based primarily on morphological criteria . These data suggest that particular subsets of Acanthamoeba strains are virulent in the human cornea.

Br J Pharmacol, 1995 Feb, 114(3), 689 - 93
Sequential induction of nitric oxide synthase by Corynebacterium parvum in different organs of the mouse; Rees DD et al.; 1 . The ability of Corynebacterium parvum (C . parvum) to induce nitric oxide (NO) synthase in the macrophage, spleen, liver, aorta, heart and brain, and to elevate plasma NO2-/NO3- in the mouse was investigated . In addition, the relationship between NO synthase activity and blood pressure was studied . 2 . C . parvum (100 mg kg-1, i.p.) induced a time-dependent expression of a Ca(2+)-independent NO synthase in the macrophage, spleen, liver, aorta and heart . The time course of induction of the NO synthase varied such that the maximum enzyme activity was at day 8 in the macrophage and liver, day 12 in the spleen and heart and day 16 in the aorta . 3 . There was no significant induction of a Ca(2+)-independent NO synthase in the brain, nor was there any change in the Ca(2+)-dependent enzyme in this organ, during the study period . 4 . C . parvum produced a gradual decrease in blood pressure, with a maximum fall at day 16 (from 108 +/- 1 mmHg to 79 +/- 3 mmHg), which recovered gradually by day 28 . 5 . Plasma NO2-/NO3- was significantly elevated between days 8 and 24, with a maximum increase at day 12 . 6 . These results show that C . parvum induces a Ca(2+)-independent NO synthase in a number of tissues and that this induction occurs initially in macrophages and the liver . This suggests that induction of the NO synthase in the other tissues is secondary and probably the result of activation of macrophages and some cells of the liver.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Epidemiol, 1995 Feb, 11(1), 107 - 17
Diphtheria: changing patterns in the developing world and the industrialized world; Galazka AM et al.; In the past, diphtheria was considered one of the most serious childhood diseases because it took a heavy toll in health and life among preschool-aged children . Prior to the widespread availability of diphtheria toxoid, nearly 70% of cases were in children younger than 15 years of age . In the industrialized countries, immunization against diphtheria became widespread in the 1940s and 1950s . This led to a marked decrease in the incidence of diphtheria . There was also a decrease in circulating toxigenic Corynebacterium diphtheriae organisms, resulting in less natural boosting of antibody levels . This had led to gaps in the immunity of the adult population . Since 1990, diphtheria has made a spectacular comeback in several European countries, with a high proportion of cases in adults . In developing countries, immunization of infants with diphtheria toxoid was introduced with the Expanded Programme on Immunization in the late 1970s . Coverage rose slowly to 46% in 1985 and 79% in 1992 . Because the pool of immunized persons is not yet large, the process of maintaining immunity still operates through natural mechanisms, including frequent skin infections caused by C . diphtheriae . But recently, several developing countries where coverage has been high for 5-10 years have reported diphtheria outbreaks . These outbreaks have been characterized by high case fatality rates, a large proportion of patients with complications, and their occurrence in both young and older age groups . In all countries, priority should be given to efforts to reach at least 90% coverage with three doses of diphtheria toxoid in children below one year of age . In countries where diphtheria has been successfully controlled, immunity levels should be maintained by booster doses.

Eur J Biochem, 1995 Jan 15, 227(1-2), 488 - 93
13C-NMR studies of Corynebacterium melassecola metabolic pathways; Rollin C et al.; Coryneform bacteria are widely used to produce amino acids, in particularly glutamic acid, by fermentation . To study the metabolic fate of glucose as the carbon source, we developed a method to analyze intracellular extracts by NMR and HPLC . The intracellular metabolites represent the metabolic state of the cells . Glutamic acid was the major metabolic intermediate found in the extracts and its 13C isotopic enrichment reflected that of pyruvic acid . Thus, it was possible to determine the respective contributions of the two major glucose catabolic pathways during the exponential growth phase; glycolysis (55%) and the pentose phosphate pathway (45%) . Absolute glutamate 13C enrichments resulting from the incorporation of {1-13C}glucose were determined to quantify the contribution of several metabolic pathways such as anaplerotic pathways (61%; phosphoenolpyruvate carboxylase, pyruvate carboxylase, malic enzyme), a single turn (32%) or multiple turns of the Krebs cycle and the glyoxylate shunt, to oxaloacetate synthesis . A previously described model was adapted to C . melassecola for these calculations . The Krebs cycle was active, whereas the glyoxylate shunt was inactive in exponentially growing cells of C . melassecola with glucose as the sole carbon source . The contributions of anaplerotic enzymes and pyruvate dehydrogenase to replenishing the Krebs' cycle were determined to be 38% and 62%, respectively.

Structure, 1995 Jan 15, 3(1), 87 - 100
Three-dimensional structure of the diphtheria toxin repressor in complex with divalent cation co-repressors; Qiu X et al.; BACKGROUND: When Corynebacterium diphtheriae encounters an environment with a low concentration of iron ions, it initiates the synthesis of several virulence factors, including diphtheria toxin . The diphtheria toxin repressor (DtxR) plays a key role in this iron-dependent, global regulatory system and is the prototype for a new family of iron-dependent repressor proteins in Gram-positive bacteria . This study aimed to increase understanding of the general regulatory principles of cation binding to DtxR . RESULTS: The crystal structure of dimeric DtxR holo-repressor in complex with different transition metals shows that each subunit comprises an amino-terminal DNA-binding domain, an interface domain (which contains two metal-binding sites) and a third, very flexible carboxy-terminal domain . Each DNA-binding domain contains a helix-turn-helix motif and has a topology which is very similar to catabolite gene activator protein (CAP) . Molecular modeling suggests that bound DNA adopts a bent conformation with helices alpha 3 of DtxR interacting with the major grooves . The two metal-binding sites lie approximately 10 A apart . Binding site 2 is positioned at a potential hinge region between the DNA-binding and interface domains . Residues 98-108 appear to be crucial for the functioning of the repressor; these provide four of the ligands of the two metal-binding sites and three residues at the other side of the helix which are at the heart of the dimer interface . CONCLUSIONS: The crystal structure of the DtxR holorepressor suggests that the divalent cation co-repressor controls motions of the DNA-binding domain . In this way the metal co-repressor governs the distance between operator recognition elements in the two subunits and, consequently, DNA recognition.

Arch Biochem Biophys, 1995 Jan 10, 316(1), 30 - 7
Targets of nitric oxide in a mouse model of liver inflammation by Corynebacterium parvum; Chamulitrat W et al.; Treatment of mice with Corynebacterium parvum induces chronic inflammation . This treatment followed by an injection of lipopolysaccharide (LPS) produces hepatic necrosis and death . We examined liver tissue by using electron paramagnetic resonance (EPR) spectroscopy and found that, in addition to the previously reported nonheme nitrosyl complexes, heme nitrosyl complexes were also formed . Hemoglobin nitrosyl complexes measured in the whole blood of mice treated with C . parvum were not increased after additional LPS treatment . However, this treatment significantly increased the heme nitrosyl complexes in the liver, whereas the nonheme nitrosyl complex concentration was unaffected . EPR signals from whole blood and liver tissues from mice treated with C . parvum and C . parvum + LPS were inhibited by prolonged treatment with NG-monomethyl-L-arginine (L-NMA) . Nitric oxide (.NO) is known to bind to cytochrome P450 heme, and we consistently found a suppression of EPR signals attributable to ferric low-spin cytochrome P450/P420 peaks in the livers of mice treated with C . parvum and C . parvum + LPS . By performing analyses of EPR spectra obtained from hepatocytes exposed to .NO, we were able to unambiguously identify EPR signals attributable to cytochrome P420 and nonheme nitrosyl complexes in the livers of both treatments . Deconvolution of the composite in vivo EPR spectra indicated that hemoglobin nitrosyl complexes contributed weakly in the C . parvum livers, but threefold more in the C . parvum + LPS livers, suggesting that hemorrhage may have occurred . Experiments with L-NMA treatment revealed that this additional .NO production did not correlate with hepatic necrosis and onset of death . Immunoprecipitation of liver cytosols from C . parvum- and (C . parvum + LPS)-treated mice using an antibody against mouse inducible nitric oxide synthase showed that this enzyme was indeed present in the cytosolic fractions and was absent in those from control livers . Our novel detection of cytochrome P420 nitrosyl complex in vivo may be linked to any role of hepatic P450's functions during liver inflammation.

Przegl Epidemiol, 1995, 49(4), 395 - 7
{A case of bacterial endocarditis (BZW) caused by Corynebacterium group JK}; Sekura-Bander D et al.; A case of bacterial endocarditis caused by Corynebacterium group JK treated with penicillin, chloramphenicol and gentamycin has been described . Finally the artificial aortal valve has been implanted.

Int Ophthalmol, 1995-96, 19(5), 313 - 6
Corynebacterium minutissimum endophthalmitis: management with antibiotic irrigation of the capsular bag; Arsan AK et al.; Chronic endophthalmitis, due to Corynebacterium minutissimum, developed in a patient following uncomplicated extracapsular cataract extraction and posterior chamber intraocular lens (PC-IOL) implantation . In this case, which to our knowledge is the first reported case of Corynebacterium minutissimum endophthalmitis, chronic inflammation persisted for 8 months with fluctuations in the inflammation . The specimens from the capsular bag yielded positive culture, but the vitreous culture was negative . The case was successfully treated by a capsular bag irrigation with vancomycin.

Pathobiology, 1995, 63(5), 270 - 7
Oxytetracycline-mediated alteration of murine immunocompetence; Myers MJ et al.; The immunomodulatory effect of oxytetracycline (OTC) on murine splenic lymphocytes (MSL), peritoneal exudate macrophage (PEM) functions and antibody production was examined . In vivo exposure to OTC slightly delayed initiation of antibody formation during the primary response . However, OTC exposure had no effect on either the peak time of antibody response or peak antibody titer . OTC also had no significant effect on the secondary antibody response . Mitogen-induced proliferation of MSL cocultured with OTC and pokeweed mitogen, phytohemagglutinin or concanavalin was equivocal . However, allogeneic stimulation of MSL was inhibited at 100 micrograms/ml OTC . There was also a decrease in the number of cells recovered . OTC had no effect on lymphocyte cytotoxicity in cells cultured in vitro . OTC inhibited the cytotoxic response of Corynebacterium parvum-elicited PEM at 10 micrograms/ml (effector:target of 10:1) . Low levels of OTC (1-10 micrograms/ml) augmented the cytotoxic response (effector:target of 5:1) . The effect of OTC on induction of PEM cytotoxicity was assessed by coculturing thioglycollate-elicited (TG) PEM in vitro with IFN-gamma and endotoxin along with 0-100 micrograms/ml OTC . Induction of cytotoxicity was inhibited at 0.5 microgram/ml . The effect of OTC on TG-PEM antimicrobial activity was assessed by measuring reduction of nitroblue tetrazolium (NBT) and cytochrome C . OTC inhibited the reduction of NBT at 500 micrograms/ml following PMA stimulation of TG-PEM . OTC had no effect on either NBT or cytochrome C reduction following stimulation with opsonized zymosan . These results demonstrate that OTC-mediated immunosuppression is a multifaceted event, with differing sensitivities both between immune cells and between different pathways within the same cell.

Acta Microbiol Immunol Hung, 1995, 42(4), 427 - 33
Role of nitrogen fixing bacteria on the phyllosphere of wheat seedlings; Pati BR et al.; Two diazotrophic bacteria Azotobacter chroococcum (REN2) and Corynebacterium sp . (Pot N2) isolated from rice and potato phyllosphere, respectively, were tested on wheat seedlings in vitro, for their efficiencies to increase dry weight and total nitrogen content of the plant, to establish their utility as potential biofertilizers . An average increase in dry weight nearly by 35-50% and total nitrogen content by 56-52% was obtained which was near to that available by nitrogenous chemical treatment . A comparatively better result was obtained by REN2 than Pot N2.

Scand J Infect Dis, 1995, 27(6), 637 - 9
Corynebacterium afermentans subsp . lipophilum: multiple abscess formation in brain and liver; Dykhuizen RS et al.; Corynebacterium afermentans, a species recently identified, has previously been isolated from human blood cultures . We report the case of a previously healthy 39-year-old man who developed a brain abscess and a liver abscess due to Corynebacterium afermentans subsp . lipophilum . The liver abscess penetrated through the diaphragm to cause pleural effusion and periostitis of the ribs . We believe this is the first reported case of disseminated infection with abscess formation due to this organism.

Scand J Infect Dis, 1995, 27(6), 635 - 6
Total knee arthroplasty complicated by Corynebacterium jeikeium infection; Yildiz S et al.; Corynebacterium jeikeium is a slow-growing multiresistant micro-organism . It causes serious opportunistic infections in patients with long hospital stays who receive broad-spectrum antibiotics . It can be isolated from blood and other sites in neutropenic, immunosuppressed bone marrow recipients and in patients with indwelling catheters . Cases of infection related to bone and joint replacements are rare . This case is the first where Corynebacterium jeikeium has been isolated from an infected knee following total knee arthroplasty.

Scand J Infect Dis, 1995, 27(6), 581 - 4
Corynebacterium CDC group JK (Corynebacterium jeikeium) sepsis in haematological patients: a report of three cases and a systematic literature review; van der Lelie H et al.; We describe 3 patients with Corynebacterium jeikeium sepsis in neutropenic phase during treatment for acute myeloid leukaemia . Fever was the first symptom . All had a central venous catheter which was removed . Two patients developed subcutaneous nodules containing pus when the neutrophil count recovered; 1 had intracutaneous and pulmonary lesions . They were treated with vancomycin and recovered when the neutrophil count started to rise . A review of 80 neutropenic patients with C . jeikeium sepsis reported in the literature, together with our 3 cases indicates that risk factors for infection are the presence of a central venous catheter, being an adult male or postmenopausal female, profound and prolonged neutropenia and exposure to multiple antibiotics . Skin lesions are reported in 48% and pulmonary lesions in 36% of the patients . The overall mortality is 34% but in patients with recovery of the bone marrow only 5% . Therefore haematopoietic growth factors should be considered in neutropenic patients with C . jeikeium infection.

Chin J Biotechnol, 1995, 11(3), 193 - 8
The factors affecting transformation efficiency of coryneform bacteria by electroporation; Na S et al.; High-voltage electroporation was performed to transfer plasmid DNA of pXZ10145 into different kinds of corynebacteria strains . A number of factors that affected transformation efficiency were investigated . Cells grown in the presence of 4% glycine and harvested in the early exponential growth phase (OD600 was about 0.2) were much more easily transformed . Transformation efficiency up to 8 x 10(6) transformants per microgram of plasmid DNA with homologously derived DNA was obtained . If heterologously derived DNA was used, transformation efficiency was 10(2) to 10(3) times lower than the former one.

Biodegradation, 1995, 6(4), 275 - 81
Effects of organic compounds on the degradation of p-nitrophenol in lake and industrial wastewater by inoculated bacteria; Zaidi BR et al.; Many microorganisms fail to degrade pollutants when introduced in different natural environments . This is a problem in selecting inocula for bioremediation of polluted sites . Thus, a study was conducted to determine the success of four inoculants to degrade p-nitrophenol (PNP) in lake and industrial wastewater and the effects of organic compounds on the degradation of high and low concentrations of PNP in these environments . Corynebacterium strain Z4 when inoculated into the lake and wastewater samples containing 20 micrograms/ml of PNP degraded 90% of PNP in one day . Addition of 100 micrograms/ml of glucose as a second substrate did not enhance the degradation of PNP and the bacterium utilized the two substrates simultaneously . Glucose used at the same concentration (100 micrograms/ml), inhibited degradation of 20 micrograms of PNP in wastewater by Pseudomonas strain MS . However, glucose increased the extent of degradation of PNP by Pseudomonas strain GR . Phenol also enhanced the degradation of PNP in wastewater by Pseudomonas strain GR, but had no effect on the degradation of PNP by Corynebacterium strain Z4 . Addition of 100 micrograms/ml of glucose as a second substrate into the lake water samples containing low concentration of PNP (26 ng/ml) enhanced the degradation of PNP and the growth of Corynebacterium strain Z4 . In the presence of glucose, it grew from 2 x 10(4) to 4 x 10(4) cells/ml in 3 days and degraded 70% of PNP as compared to samples without glucose in which the bacterium declined in cell number from 2 x 10(4) to 8 x 10(3) cells/ml and degraded only 30% PNP . The results suggest that in inoculation to enhance biodegradation, depending on the inoculant, second organic substrate many play an important role in controlling the rate and extent of biodegradation of organic compounds.

Rev Elev Med Vet Pays Trop, 1995, 48(2), 133 - 7
{Main infectious agents involved in the etiology of lung diseases of small ruminants in northern Cameroon}; Martrenchar A et al.; Between 1990 and 1992, 91 necropsies of small ruminants affected with pulmonary illness led to the isolation of the following strains of Mycoplasma (M.): M . mycoides subsp . mycoides LC, M . ovipneumoniae, M . agalactiae, M . sp . type D2 and M . arginini . Eleven Pasteurella multocida strains (serotypes A1, A3, A5, A7 and D2) and 11 Pasteurella haemolytica strains (serotypes 1, 2, 3, 6, 7, 8 and 9) were isolated . Corynebacterium pseudotuberculosis, Actinomyces pyogenes, Staphylococcus sp., Streptococcus sp., Bacillus sp . and Mycobacterium sp . were also isolated . Thirty-two antibiograms were performed on Pasteurella, Corynebacterium pseudotuberculosis and Actinomyces pyogenes strains . Eighty eight p . cent were sensitive to penicillin G and oxytetracycline, and 84% to chloramphenicol; 50% were not sensitive to spiramycin and 47% to streptomycin . One Capripoxvirus strain was isolated on sheep . Pest of small ruminants (PPR) virus was detected by immunocapture ELISA test performed on some lung samples . Two serological surveys, one for contagious caprine pleuropneumonia (898 goats), between 1991 and 1993, and one for PPR (902 sheep and goats) in 1993, were conducted in the North and Far North provinces . No antibody against contagious caprine pleuropneumonia was detected . Among the animals in the sample, PPR prevalence was 64 +/- 7% in the Far North province and 14 +/- 3% in the North province . Concerning control measures, a vaccination campaign against small ruminant pasteurellosis appears to be hardly feasible because of the antigenic diversity of the isolated Pasteurella strains . PPR is endemic especially in the Far North province . The efficiency of a vaccination campaign against PPR must be estimated with a field survey.

Appl Environ Microbiol, 1995 Jan, 61(1), 74 - 8
Effect of inducible thrB expression on amino acid production in Corynebacterium lactofermentum ATCC 21799; Colon GE et al.; Amplification of the operon homdr-thrB encoding a feedback-insensitive homoserine dehydrogenase and a wild-type homoserine kinase in a Corynebacterium lactofermentum lysine-producing strain resulted in both homoserine and threonine accumulation, with some residual lysine production . A plasmid enabling separate transcriptional control of each gene was constructed to determine the effect of various enzyme activity ratios on metabolite accumulation . By increasing the activity of homoserine kinase relative to homoserine dehydrogenase activity, homoserine accumulation in the medium was essentially eliminated and the final threonine titer was increased by about 120% . Furthermore, a fortuitous result of the cloning strategy was an unexplained increase in homoserine dehydrogenase activity . This resulted in a further decrease in lysine production along with a concomitant increase in threonine accumulation.

Arch Bronconeumol, 1995 Jan, 31(1), 40 - 2
{Pleural empyema caused by Corynebacterium sp.}; Carrion F et al.; Until recently Corynebacterium sp . had been considered an infrequent cause of infection in humans . In the last few years, however, it has proven its ability to produce severe pulmonary and extrapulmonary infections, generally in immunodeficient hosts . We present a rare case of an immunocompetent man 68 years-old who developed community-acquired pneumonia and left pleural empyema, with Corynebacterium sp . isolated in pleural liquid . Response to thoracic drainage and antibiotic therapy (imipenem and clindamycin) was good.

Acta Oncol, 1995, 34(1), 117 - 21
Pleurodesis with doxycycline or Corynebacterium parvum in malignant pleural effusion; Salomaa ER et al.; Pleurodesis with doxycycline (100 mg and 600 mg) and Corynebacterium parvum (1 mg and 7 mg) were compared in 41 patients with malignant effusion . To evaluate the mechanisms, pleural fluid pH, leukocytes, granulocytes, interleukin-6 (IL-6) and serum IL-6, as well as C-reactive protein (CRP) were measured before and on 2 consecutive days after treatment . Corynebacterium parvum produced a greater acute-phase response measured with fever, serum CRP and IL-6 than doxycycline . However, no change in pleural fluid IL-6 was demonstrated . Among the 35 assessed patients, 26 had objective response, similar in all four treatment groups . Side-effects were more common with Corynebacterium parvum . Based on this preliminary study we conclude that doxycycline, even in low doses, is a highly effective and well tolerated agent for palliative treatment of malignant pleural effusion . As the responses were similar despite different inflammatory reactions, the two agents probably induce pleural obliteration through different mechanisms.

Int J Syst Bacteriol, 1995 Jan, 45(1), 32 - 6
Dietzia, a new genus including Dietzia maris comb . nov., formerly Rhodococcus maris; Rainey FA et al.; Sequencing of the 16S ribosomal DNAs (rDNA) of two strains of Rhodococcus maris was performed to determine the relationship of this species to other mycolic acid-containing actinomycetes . For this purpose we also determined the 16S rDNA sequences for the type species of the genus Rhodococcus, Rhodococcus rhodochrous, and for Mycobacterium chlorophenolicum (formerly Rhodococcus chlorophenolicus), Rhodococcus erythropolis, Gordona bronchialis, and Gordona terrae, for which only partial sequence data have been available previously . The sequences of the two strains of R . maris were identical . The results of a distance matrix analysis indicated that R . maris is not a member of the genus Rhodococcus but is located between members of the genus Corynebacterium and members of the Rhodococcus-Nocardia-Mycobacterium-Gordona-Tsukamurella cluster . The finding that R . maris is phylogenetically isolated is supported by the presence of N-acetyl residues in the glycan moiety of the peptidoglycan and the lack of phosphatidylinositol and phosphatidylinositol mannosides, characteristics which distinguish this taxon from related taxa . On the basis of our results and previous findings, we propose that R . maris should be reclassified in a new genus, Dietzia . The type species is Dietzia maris comb . nov.

Int J Syst Bacteriol, 1995 Jan, 45(1), 128 - 33
Genomic diversity and phylogenetic relationships among lipid-requiring diphtheroids from humans and characterization of Corynebacterium macginleyi sp . nov; Riegel P et al.; DNA relatedness experiments were performed with 38 clinical isolates and 13 reference strains of coryneform taxa exhibiting a lipid requirement for optimal growth . Forty-five of these strains split into five genomic groups at the species level, whereas six other strains remained unclustered . Genomospecies II fits Corynebacterium accolens, but the other genomospecies were not genetically related to any of the defined Corynebacterium species . Phylogenetic analyses of genes coding for small-subunit rRNA sequences revealed that two genomospecies (I and III) and C . accolens form a tight cluster within the robust branch that groups all Corynebacterium species presently sequenced . Reference strains of biotypes C-1, C-2, and C-3 of "Corynebacterium pseudogenitalium" were found to fall into genomospecies I, as well as "Corynebacterium tuberculostearicum," Centers for Disease Control and Prevention (CDC) coryneform group G-1, and CDC coryneform group G-2 reference strains . Biochemical tests allowed differentiation between genomospecies except between genomospecies IV and V and between six unclustered strains and genomospecies I . We propose a new classification for these lipid-requiring diphtheroids within the genus Corynebacterium with the delineation of some CDC coryneform group G-1 strains (genomospecies III) as a new species for which the name Corynebacterium macginleyi is proposed . The type strain is strain JCL-2 (CIP 104099), isolated from a human corneal ulcer.

Dig Dis Sci, 1995 Jan, 40(1), 41 - 7
Use of prostaglandin I2 analog in treatment of massive hepatic necrosis associated with endothelial cell injury and diffuse sinusoidal fibrin deposition; Fujiwara K et al.; Endothelial cell damage causes massive hepatic necrosis as a result of fibrin deposition in the hepatic sinusoids . When a stable analog of prostaglandin I2, beraprost sodium, was administered to rats given either dimethylnitrosamine, carbon tetrachloride, or endotoxin following Corynebacterium parvum administration, the hepatic necrosis produced in each was attenuated, but to a greater extent in the dimethylnitrosamine and endotoxin/Corynebacterium parvum models, where fibrin deposition in the hepatic sinusoids occurs, as compared to the carbon tetrachloride model, where such fibrin deposition does not occur . Beraprost sodium reduced the expected increase of portal venous pressure in the endotoxin/Corynebacterium parvum model without affecting plasma thrombin-antithrombin III complex levels . Beraprost sodium also significantly reduced cell killing of both isolated rat hepatocytes and hepatic sinusoidal endothelial cells exposed to tert-butyl hydroperoxide when compared to controls . Beraprost sodium could prove to be a therapeutic candidate for the treatment of hepatic necrosis, particularly in cases associated with fibrin deposition in the hepatic sinusoids because of its fibrin clot-clearing action.

Infect Immun, 1995 Jan, 63(1), 206 - 11
Molecular and biochemical characterization of a protective 40-kilodalton antigen from Corynebacterium pseudotuberculosis; Wilson MJ et al.; A 40-kDa protein from Corynebacterium pseudotuberculosis has been previously identified as a protective antigen against ovine caseous lymphadenitis . From genomic DNA libraries of C . pseudotuberculosis, we have cloned and sequenced the 40-kDa protein gene, which was found to contain an open reading frame of 1,137 bp encoding a protein of 379 amino acids . No significant homology with previously published DNA or amino acid sequence data was found in databases, suggesting that this is a novel protein . Recombinant 40-kDa protein was overexpressed as a fusion protein to 15% of total cell proteins in Escherichia coli . Biochemical analysis of native and recombinant 40-kDa proteins has revealed associated proteolytic activity, which was shown to be of the serine protease type through the use of specific inhibitors . We suggest that this novel protective antigen be termed corynebacterial protease 40 (CP40).

Diagn Microbiol Infect Dis, 1995 Jan, 21(1), 55 - 6
Comparison of one-day versus two-day incubation of urine cultures; Joho KL et al.; The value of incubating urine cultures for 1 versus 2 days was evaluated prospectively for 1526 consecutive specimens . A total of 507 cultures (33.2%) were positive after 1 day; 41 (2.7%) showed different results after 2 days . Only yeasts and corynebacteria were detected more often with longer incubation . Patient charts were available for review from 27 of 41 late positives; in only three instances (11.1%) was action taken by physicians based on these results.

Antonie Van Leeuwenhoek, 1995, 67(2), 221 - 7
Purification and properties of oxaloacetate decarboxylase from Corynebacterium glutamicum; Jetten MS et al.; Oxaloacetate (OAA) decarboxylase (E.C . 4.1.1.3) was isolated from Corynebacterium glutamicum . In five steps the enzyme was purified 300-fold to apparent homogeneity . The molecular mass estimated by gel filtration was 118 +/- 6 kDa . SDS-PAGE showed a single subunit of 31.7 KDa, indicating an alpha 4 subunit structure for the native enzyme . The enzyme catalyzed the decarboxylation of OAA to pyruvate and CO2, but no other alpha-ketoacids were used as substrate . The cation Mn2+ was required for full activity, but could be substituted by Mg2+, CO2+, Ni2+ and Ca2+ . Monovalent ions like Na+, K+ or NH4+ were not required for activity . The enzyme was inhibited by Cu2+, Zn2+, ADP, coenzyme A and succinate . Avidin did not inhibit the enzyme activity, indicating that biotin is not involved in decarboxylation of OAA . Analysis of the kinetic properties revealed a Km for OAA of 2.1 mM and a Km of 1.2 mM for Mn2+ . The Vmax was 158 mumol of OAA converted per min per mg of protein, which corresponds to an apparent kcat of 311 s-1.

Appl Microbiol Biotechnol, 1995 Jan, 42(5), 724 - 9
Cloning of FAD synthetase gene from Corynebacterium ammoniagenes and its application to FAD and FMN production; Hagihara T et al.; The cloning of a bifunctional FAD synthetase gene, which shows flavokinase and FMN adenylyltransferase activities, from Corynebacterium ammoniagenes was tried by hybridization with synthetic DNAs corresponding to the N-terminal amino acid sequence . The cloned PstI-digested 4.4 x 10(3)-base (4.4-kb) fragment could not express the FAD synthetase activity in E . coli, but could increase the two activities by the same factor of about 20 in C . ammoniagenes . The FAD-synthetase-gene-amplified C . ammoniagenes cells were applied to the production of FAD from FMN or riboflavin . The productivity of FAD from FMN was increased four to five times compared with the parent strain, and reached a 90% molar yield . The productivity of FAD from riboflavin was increased about eight times, with a 50% molar yield . The addition of Zn2+ to the reaction mixtures for the conversion from riboflavin to FAD brought about the specific inhibition of adenylyl-transferase activity and resulted in the accumulation of FMN.

Vestn Ross Akad Med Nauk, 1995, (2), 47 - 51
{Search, isolation and study of restrictases}; Sokolov NN; New Class II restrictases were searched in over 800 microorganisms by using a highly sensitive toluene microtechnique developed in the laboratory . This enabled site-specific endonuclease activity to be revealed in 72 strains . Thirty two new restriction endonucleases were identified, which were highly purified and contained no impurities of nonspecific nucleases, phosphatases . Many of them (LpII, PaeI, PaeBI, ApiI, CsiAI, BavAI, BavAIII, BbvAI, etc.) are of interest for use in molecular genetic studies . Corynebacterium species cells were used to isolate a new supercoarse hissing restrictase CsiBI that recognizes the 8-nucleotide site GGGGGGGG (the isoschizomer NotI) and a fine tool for obtaining enlarged fragments of pro- and eukaryotic genome.

Kansenshogaku Zasshi, 1995 Jan, 69(1), 105 - 13
{Yearly changes of isolated organisms from the respiratory tract in Hokusho Central Hospital}; Takase T et al.; Isolated organisms from the respiratory tract have been studied in our hospital from 1986 to 1993 . The total number of samples were 18,345 and samples which showed 10(5) cfu/ml organisms were 8648 in our hospital for 8 years . Enterobacteriacae, Pseudomonas aeruginosa, Haemophilus influenzae, Streptococcus pneumoniae, and glucose nonfermenting gram-negative rods were major isolates in 8 years . Haemophilus influenzae, which used to be the commonest isolate, decreased from 10.9% in 1993 while Enterobacteriacae increased from 8.9% in 1986 to 17.6% in 1993 . S . pneumoniae and H . influenzae were major isolates from out-patients consisting of 50%, followed by Enterobacteriacae, P . aeruginosa and MSSA . Enterobacteriacae and P . aeruginosa were major isolates from in-patients, followed by MRSA and beta-Streptococcus . Streptococcus agalactiae, Serratia marcescens and Corynebacterium spp . prevailed especially in the geriatric ward . S . pneumoniae, H . influenzae and M . catarrhalis were major isolates from patients with pneumoconiosis, especially in winter.

Crit Rev Biotechnol, 1995, 15(1), 73 - 103
Recent advances in the physiology and genetics of amino acid-producing bacteria; Jetten MS et al.; Corynebacterium glutamicum and its close relatives, C . flavum and C . lactofermentum, have been used for over 3 decades in the industrial production of amino acids by fermentation . Since 1984, several research groups have started programs to develop metabolic engineering principles for amino acid-producing Corynebacterium strains . Initially, the programs concentrated on the isolation of genes encoding (deregulated) biosynthetic enzymes and the development of general molecular biology tools such as cloning vectors and DNA transfer methods . With most of the genes and tools now available, recombinant DNA technology can be applied in strain improvement . To accomplish these improvements, it is critical and advantageous to understand the mechanisms of gene expression and regulation as well as the biochemistry and physiology of the species being engineered . This review explores the advances made in the understanding and application of amino acid-producing bacteria in the early 1990s.

Clin Infect Dis, 1995 Jan, 20(1), 41 - 6
Corynebacterium pseudodiphtheriticum: a respiratory tract pathogen; Ahmed K et al.; From January 1986 through February 1993, there were 16 episodes of respiratory infection due to Corynebacterium pseudodiphtheriticum in 13 patients . The ages of patients ranged from 24 to 77 years; the ratio of male to female patients was 3:1 . One patient had three episodes of infection, and another patient had two . In one patient, who died of disseminated intravascular coagulation, the level of IgG was low . One patient was receiving prednisolone when the infection occurred . In two cases a mixed infection with Streptococcus pneumoniae was noted . Sputum cultures yielded C . pseudodiphtheriticum (> or = 10(7) cfu/mL) . An increased neutrophil response in the sputum of infected patients was observed . Gram staining and electron microscopy of sputum showed phagocytosis of C . pseudodiphtheriticum by the neutrophils . ELISAs also showed an increase in the level of immunoglobulin against C . pseudodiphtheriticum after infection . Tests for determination of MICs of antibiotics revealed that C . pseudodiphtheriticum isolates were susceptible to ampicillin, amoxycillin/clavulanic acid, cefazolin, cefuroxime, ceftazidime, and imipenem . All strains were resistant to nalidixic acid; borderline susceptibility to ofloxacin, norfloxacin, and ciprofloxacin was noted . We suggest the use of beta-lactam antibiotics in the treatment of infection with C . pseudodiphtheriticum.

Clin Infect Dis, 1995 Jan, 20(1), 37 - 40
Corynebacterium pseudodiphtheriticum: a respiratory tract pathogen in adults; Manzella JP et al.; Corynebacterium pseudodiphtheriticum has been reported to be an uncommon respiratory pathogen . We describe the clinical and microbiologic features of 17 patients from whose sputum C . pseudodiphtheriticum was isolated . Patients were identified through a review of the reports from the clinical microbiology laboratory at York Hospital, a community teaching hospital, from October 1990 through April 1993; 17 patients with respiratory infection caused by C . pseudodiphthriticum were identified . There were 12 cases of bronchitis and five of pneumonia . An underlying systemic condition, particularly congestive heart failure, chronic obstructive pulmonary disease, diabetes mellitus, or malignancy, was common . Onset of symptomatology was acute for most patients, but fever was noticeably absent in almost two-thirds of the cases . Isolates were uniformly susceptible to the beta-lactam antibiotics, vancomycin, and trimethoprim-sulfamethoxazole, but resistance to clindamycin and erythromycin was common . The isolation of diphtheroids from a properly obtained sputum sample from a patient with respiratory tract infection should not always be dismissed as due to contamination . The isolation, identification, and susceptibility testing of C . pseudodiphtheriticum from respiratory tract specimens may provide information useful for treatment of patients.

Vet Pathol, 1995 Jan, 32(1), 68 - 71
Local production of tumor necrosis factor-alpha in corynebacterial pulmonary lesions in sheep; Ellis JA et al.; Corynebacterium pseudotuberculosis infection is a common cause of pyogranulomas in ovine lungs and often occurs as a dual infection with lentiviruses . This coinfection usually leads to the development of chronic pneumonia and cachexia that is similar to the clinical syndrome seen in human beings with AIDS-related pneumonias . Recent in vitro studies indicate that monokines such as tumor necrosis factor-alpha (TNF alpha) are induced by C . pseudotuberculosis, suggesting that TNF alpha is involved in the pathogenesis of corynebacterial lesions in vivo . To substantiate in vitro observations concerning bacterial induction of TNF alpha in ovine pulmonary macrophages, immunohistochemical labeling techniques were used in combination with in situ hybridization to identify TNF-producing cells in corynebacterial lesion sites in vivo . TNF alpha message and translation product were found in macrophages comprising pyogranulomas that were induced by naturally acquired and experimental pulmonary C . pseudotuberculosis infections.

Respiration, 1995, 62(1), 21 - 6
Intrapleural Corynebacterium parvum for recurrent malignant pleural effusions; Foresti V; Twenty-two consecutive patients with malignant pleural effusions (MPE) were treated with intrapleural Corynebacterium parvum (CBP) associated with parenteral methylprednisolone (MP) to determine its effectiveness and the frequency and nature of adverse reactions . After thoracentesis, 7 mg of CBP (Coparvax Wellcome) in 20 ml of saline were injected into the pleural cavity . On the day of treatment, the patients were given 1 mg/kg i.m . of MP 30 min before thoracentesis . The effectiveness of pleurodesis was assessed as follows: (1) complete response (CR; total resolution of pleural effusion after 3 injections of CBP at the most); (2) partial response (PR; formation of asymptomatic loculated effusion) . In 5 patients leukocytes, lymphocytes and monocytes were determined in pleural fluid (PF) and in blood (B) collected before and 7 days after CBP treatment . Two patients were unevaluable . Of 20 evaluable patients, 18 (90%) had a CR and 2 patients (10%) had a PR . Eleven of 22 patients (50%) had a fever . Three patients had prolonged and/or high fever . Seven of 22 patients (32%) had mild chest pain . None of the patients presented other side effects . Twelve of 21 patients (57.1%) had a PF pH > or = 7.30; 2 of these died a few days after the treatment, and 10 had favorable responses . The other 9 patients had a PF pH < 7.30: all had favorable responses . The leukocytes, the lymphocyte subsets, the monocytes, the NK lymphocytes, and their PF/B ratios did not differ significantly before and after CBP treatment . Our study confirms that intrapleural CBP is an effective and simple method to control MPE.(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1995 Jan, 39(1), 208 - 14
Antimicrobial susceptibilities of Corynebacterium species and other non-spore-forming gram-positive bacilli to 18 antimicrobial agents; Soriano F et al.; The susceptibilities of 265 strains of Corynebacterium species and other non-spore-forming gram-positive bacilli to 18 antimicrobial agents were tested . Most strains were susceptible to vancomycin, doxycycline, and fusidic acid . Corynebacterium jeikeium and Corynebacterium urealyticum were the most resistant organisms tested . Resistance to beta-lactams, clindamycin, erythromycin, azythromycin, ciprofloxacin and gentamicin was common among strains of Corynebacterium xerosis and Corynebacterium minutissimum . Ampicillin resistance among Listeria monocytogenes was more prevalent than previously reported . Optochin, fosfomycin, and nitrofurantoin showed very little activity against most organisms tested, but the use of nitrofurantoin as a selective agent in culture medium may prevent the recovery of some isolates . Except for the unvarying activity of vancomycin against Corynebacterium species, the antimicrobial susceptibilities of the latter to other antibiotics are usually unpredictable, such that susceptibility tests are necessary for selecting the best antimicrobial treatment.

Pathology, 1995 Jan, 27(1), 71 - 3
Polymerase chain reaction for the detection of toxigenic Corynebacterium diphtheriae; Aravena-Roman M et al.; Several conventional methods have been described for the detection of Corynebacterium diphtheriae toxin, including Elek immunodiffusion, tissue culture using VERO cells and guinea pig inoculation . All these methods have the disadvantage of being either slow to complete or technically demanding, particularly when performed infrequently . We examined 64 strains of C . diphtheriae by PCR and Elek immunodiffusion, and strains showing a positive result in either assay were inoculated into guinea pigs . Seven isolates were positive in both Elek and PCR assays and subsequently positive in guinea pig inoculation assay . One isolate was negative in Elek testing but positive in PCR assay and guinea pig inoculation . All other isolates were negative in both Elek and PCR assays . The PCR assay is rapid with cycling and detection complete within 3-4 hrs of receipt of strains . PCR has now become the routine method for detection of C . diphtheriae toxin in our laboratory.

Biol Cell, 1995, 83(2-3), 219 - 229
Organization of the outer layers of the cell envelope of Corynebacterium glutamicum: a combined freeze-etch electron microscopy and biochemical study; Chami M et al.; The cell surface of Corynebacterium glutamicum grown on solid medium was totally covered with a highly ordered, hexagonal surface layer . Also, freeze-fracture revealed two fracture surfaces which were totally covered with ordered arrays displaying an hexagonal arrangement and the same unit cell dimension as the surface layer . The ordered arrays on the concave fracture surface, closest to the cell surface, were due to the presence of particles while those on the convex fracture surface were their imprints . The same cells grown on liquid medium displayed a cell surface and fracture surfaces only partially covered with ordered arrays . In this case, the ordered regions had the same relative position on the cell surface and on the fracture surfaces . All ordered arrays were totally absent in a mutant for cspB, the gene encoding PS2, one of the two major cell wall proteins . Treatment of the cells with proteinase K caused the gradual alteration of PS2 into a slightly lower molecular mass form . This was accompanied by a concomitant disappearance of the ordered fracture surfaces followed by the detachment of the ordered surface layer from the cell as large ordered patches displaying the same lattice symmetry and dimensions as those of the surface layer . The ordered patches were isolated . They contained the totality of PS2 initially associated with the cell . We conclude that the highly ordered surface layer of the intact cell was composed of PS2 interacting strongly with some cell wall material leading to its organization . This organized cell wall material produced fracture surfaces . We show that in the absence of intact PS2 protein on the cell wall, the same cell wall material was not organized and formed a structureless smooth layer.

Bull Cancer, 1995, 82 Suppl 2, 127s - 138s
{Immunotherapy of malignant diseases}; Dorval T et al.; Cancer immunotherapy has been carried out since the early fifties and first involved nonspecific system (BCG, Corynebacterium parvum, levamisole...) . More recently, the production of cytokines as interferons or interleukin 2, the introduction of monoclonal antibodies have allowed a new development to cancer immunotherapy . Nevertheless, these new approaches have to be considered as a step in the biological therapy of cancer.

Presse Med, 1994 Dec 17, 23(40), 1859 - 61
{Non-toxic Corynebacterium diphtheriae septicemia with endocarditis in an earlier healthy adult . First case and review of the literature}; Breton D; Corynebacterium diphtheriae septicaemia is rarely encountered, usually in very particular situations: children with severe congenital heart disease or after heart surgery . Rare cases have been reported in immunodepressed adults or drug addicts . We observed a case in a formerly healthy 41-year-old woman who was hospitalized for fever unresponsive to bacampicillin . In this patient, no portal of entry could be identified; there was no history of past surgery nor drug abuse . The patient was not immunodepressed and HIV serology was negative . The last anti-diphtheria vaccination had been given at the age of 12 years . Corynebacterium diphtheriae var . metis was identified on five blood cultures . The in vitro Elek test revealed that the strain was non-toxic . Echocardiography did not show any signs until the fourth examination performed 1 month after onset of fever and 15 days after initiating effective adapted antibiotic treatment with amoxicillin-clavanic acid . Mitral vegetations with grade 2 regurgitation completely regressed after 5 weeks of treatment . After 5 months of follow-up, the patient is in good health and no mitral damage has been observed . This is to our knowledge the first case report of Corynebacterium diphtheriae in a formerly healthy adult . In the literature 12 other cases in adults all concerned immunodepressed subjects or drug abusers . The question is raised as to whether Corynebacterium diphtheriae is undergoing mutation . The germ could persist as a commensal host and explain a certain number of the recent observations in drug abusers and immunodepressed patients.

J Mol Biol, 1994 Dec 16, 244(5), 654 - 6
Crystallization and preliminary X-ray studies of the diphtheria Tox repressor from Corynebacterium diphtheriae; Schiering N et al.; Crystals of the diphtheria tox repressor (DtxR) from Corynebacterium diphtheriae suitable for structure determination have been obtained . DtxR activated with transition metal ions represses the expression of the structural gene for the diphtheria toxin, tox, which is encoded on the genome of a family of closely related corynebacteriophages . The space group of the obtained crystals is trigonal P3(1)21 or its enantiomorph P3(2)21 with a = b = 64.2 A, c = 220.5 A, alpha = beta = 90 degrees, gamma = 120 degrees . Two monomers comprise the asymmetric unit . The crystals diffract to a resolution of better than 3 A.

FEBS Lett, 1994 Dec 12, 356(1), 104 - 8
Characterization of energetically functional inverted membrane vesicles from Corynebacterium glutamicum; Schrempp S et al.; We show that inverted membrane vesicles from Corynebacterium glutamicum, a Gram-positive bacterium, are able to generate and maintain an electrochemical gradient of protons in response to the addition of NADH . This result indicates that the respiratory chain is intact and that the vesicles are reasonably impermeable to protons . These membrane vesicles may be the starting point for in vitro translocation studies of proteins in Gram-positive bacteria.

Schweiz Med Wochenschr, 1994 Dec 3, 124(48), 2173 - 80
{Endocarditis due to Corynebacterium diphtheriae cause by contact with intravenous drugs: report of 5 cases}; Huber-Schneider C et al.; The clinical features of 5 patients with invasive disease due to nontoxigenic Corynebacterium diphtheriae are presented . 4 patients had proven left sided endocarditis, and one had probable endocarditis of the mitral valve . 4 patients were intravenous drug abusers, and one had a girl friend who was also an iv-drug abuser . Only one patient had antibodies against human immunodeficiency virus . 2 patients with endocarditis of the mitral or the aortic valve, and one patient with endocarditis of a prosthetic aortic valve, died of septic complications . All patients or their partners frequented the same community of drug abusers . Since the blood culture isolates shared common features for biotyping, resistance patterns and also for ribotyping in three tested patients, we assume that all patients were infected by the same clone of C . diphtheriae . Drug abusers may become a new reservoir for C . diphtheriae infection . Along with our patients, we present an overview of the 49 published cases of C . diphtheriae endocarditis.

J Bacteriol, 1994 Dec, 176(23), 7309 - 19
Cloning and characterization of a DNA region encoding a stress-sensitive restriction system from Corynebacterium glutamicum ATCC 13032 and analysis of its role in intergeneric conjugation with Escherichia coli; Schafer A et al.; RP4-mediated transfer of mobilizable plasmids in intergeneric conjugation of Escherichia coli donors with Corynebacterium glutamicum ATCC 13032 is severely affected by a restriction system in the recipient that can be inactivated by a variety of exogenous stress factors . In this study a rapid test procedure based on intergeneric conjugal plasmid transfer that permitted the distinction between restriction-negative and restriction-positive C . glutamicum clones was developed . By using this procedure, clones of the restriction-deficient mutant strain C . glutamicum RM3 harboring a plasmid library of the wild-type chromosome were checked for their restriction properties . A complemented clone with a restriction-positive phenotype was isolated and found to contain a plasmid with a 7-kb insertion originating from the wild-type chromosome . This plasmid, termed pRES806, is able to complement the restriction-deficient phenotype of different C . glutamicum mutants . Sequence analysis revealed the presence of two open reading frames (orf1 and orf2) on the complementing DNA fragment . The region comprising orf1 and orf2 displayed a strikingly low G+C content and was present exclusively in C . glutamicum strains . Gene disruption experiments with the wild type proved that orf1 is essential for complementation, but inactivation of orf2 also resulted in a small but significant increase in fertility . These results were confirmed by infection assays with the bacteriophage CL31 from Corynebacterium lilium ATCC 15990.

Infect Immun, 1994 Dec, 62(12), 5275 - 80
Protection of sheep against caseous lymphadenitis by use of a single oral dose of live recombinant Corynebacterium pseudotuberculosis; Hodgson AL et al.; An inactive form of the Corynebacterium pseudotuberculosis phospholipase D (PLD) gene was constructed and expressed in a PLD-negative strain (designated Toxminus) of C . pseudotuberculosis . Antibody responses specific to Toxminus and both Toxminus and PLD proteins were detected in sheep following oral administration of Toxminus or Toxminus expressing the PLD toxoid, respectively . However, only those sheep vaccinated with Toxminus expressing PLD toxoid were protected against wild-type challenge . These results confirm the importance of PLD as a protective antigen and demonstrate both the potential for developing an oral caseous lymphadenitis vaccine and C . pseudotuberculosis Toxminus as a live vaccine vector.

Cancer Res, 1994 Dec 1, 54(23), 6022 - 6
Immunizing and curative potential of replicating and nonreplicating murine mammary adenocarcinoma cells engineered with interleukin (IL)-2, IL-4, IL-6, IL-7, IL-10, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, and gamma-interferon gene or admixed with conventional adjuvants; Allione A et al.; To evaluate the efficacy of vaccinations with cytokine-gene-transduced tumor cells, BALB/c mice were challenged with 1 x 10(5) parental cells of a syngeneic adenocarcinoma cell line (TSA-pc) . No protection was observed in mice immunized 30 days earlier with 1 x 10(5) nonreplicating mitomycin-C-treated TSA-pc alone, or with Corynebacterium parvum or Complete Freund Adjuvant (CFA) . Ten to 30% of mice immunized with nonreplicating cells engineered to produce interleukin (IL)-2, IL-4, IL-6, IL-7, IL-10, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, and gamma-interferon gene were protected . Fifty % of mice immunized with replicating TSA-pc admixed with C . parvum and 80-100% of mice immunized with replicating tumor cells transduced with IL-2, IL-4, IL-7, IL-10, or gamma-interferon gene were protected . No cure was afforded by TSA cells admixed with C . parvum or CFA, nor by TSA cells engineered with IL-6, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor alpha gene injected starting 1 day after TSA-pc challenge . Complete tumor regression, however, was obtained in 10-20% of mice treated with TSA cells transduced with IL-2, IL-4, IL-7, or IL-10 and in 30% of those treated with TSA cells transduced with gamma-interferon gene.

Clin Infect Dis, 1994 Dec, 19(6), 1054 - 61
Native valve endocarditis due to Corynebacterium striatum: case report and review; Rufael DW et al.; We report the first known case of native valve endocarditis due to Corynebacterium striatum and review 51 previously reported cases of native valve endocarditis due to non-diphtheriae corynebacteria . Of the 52 patients with corynebacterial endocarditis, 11 (21%) had no predisposing conditions and 27 (52%) had structural heart disease; endocarditis in the remaining 14 patients (27%) was associated with noncardiac predisposing factors including injection drug use, chronic hemodialysis, vasculitis, alcoholism, liver transplantation and hemodialysis, a peritoneovenous shunt, and prior aspiration of a noninfected bursa . The mortality rate associated with corynebacterial endocarditis was 31% . The majority of corynebacteria in this series were sensitive to penicillin, erythromycin, gentamicin, and vancomycin . Non-diphtheriae corynebacteria are capable of producing acute valvular damage, even in patients without conditions that are predisposing for endocarditis . The occurrence of bacteremia due to non-diphtheriae corynebacteria in the appropriate clinical setting should alert physicians to the possible diagnosis of endocarditis . Empirical antibiotic therapy with vancomycin, with or without an aminoglycoside, should be initiated pending antibiotic susceptibility testing.

Microbiology, 1994 Dec, 140 ( Pt 12), 3349 - 56
Analysis of different DNA fragments of Corynebacterium glutamicum complementing dapE of Escherichia coli; Wehrmann A et al.; In Corynebacterium glutamicum L-lysine is synthesized simultaneously via the succinylase and dehydrogenase variant of the diaminopimelate pathway . Starting from a strain with a disrupted dehydrogenase gene, three different-sized DNA fragments were isolated which complemented defective Escherichia coli mutants in the succinylase pathway . Enzyme studies revealed that in one case the dehydrogenase gene had apparently been reconstituted in the heterologous host . The two other fragments resulted in desuccinylase activity; one of them additionally in succinylase activity . However, the physical analysis showed that structural changes had taken place in all fragments . Using a probe derived from one of the fragments we isolated a 3.4 kb BamHI DNA fragment without selective pressure (by colony hybridization) . This was structurally intact and proved functionally to result in tenfold desuccinylase overexpression . The nucleotide sequence of a 1966 bp fragment revealed the presence of one truncated open reading frame of unknown function and that of dapE encoding N-succinyl diaminopimelate desuccinylase (EC 3.5.1.18) . The deduced amino acid sequence of the dapE gene product shares 23% identical residues with that from E . coli . The C . glutamicum gene now available is the first gene from the succinylase branch of lysine synthesis of this biotechnologically important organism.

Kansenshogaku Zasshi, 1994 Dec, 68(12), 1527 - 32
{A case of Corynebacterium jeikeium septicemia}; Otsuka Y et al.; We report a case of Corynebacterium jeikeium septicemia associated with malignant lymphoma . The patient is a 58-year-old male who was diagnosed as malignant lymphoma on August 1992 . May 15, 1993, he was admitted to our hospital because of oliguria, abdominal flatulence and vomiting which developed a few days before admission . Anticancer regimen were started . In the middle of July, white blood cell (WBC) count dropped to 100/mm3 and body temperature rose to 39 degrees C . He was been treated with Ceftazidime and Piperacillin . C . jeikeium was recovered from blood culture . Antibiotics were switched to minocycline and vancomycin . He died of septic shock and pneumonia . Autopsy revealed the presence of the colonies of Rods . Which were morphologically compatible with C . jeikeium were observed in lung tissue and in the small pulmonary vessels.

Oral Microbiol Immunol, 1994 Dec, 9(6), 352 - 8
Carbohydrate depletion of immunoglobulin A1 by oral species of gram-positive rods; Frandsen EV; Bacterial deglycosylation of immunoglobulin Al (IgA1), the dominant isotype of antibody in the oral cavity, probably provides both nutrition as well as protection to the oral bacterial community . Representative strains of oral gram-positive rods were tested for their ability to remove carbohydrates from IgA1 . Detection of sialic acids was performed by means of high-performance liquid chromatography (HPLC) separation (Aminex HPX-87H) and ultraviolet light absorption at 190 nm, and neutral carbohydrates were measured by HPLC separation (Capcell Pak C-18 SG 120) and ultraviolet light absorption at 245 nm after derivatization . Four strains of Actinomyces naeslundii, two strains of Corynebacterium matruchotii and one of two strains of Actinomyces odontolyticus partially or totally removed sialic acid, while two strains of Propionibacterium propionicus and the other strain of A . odontolyticus did not . Complete correlation was observed between sialic acid removal, neuraminidase activity measured with fluorogenic substrate and with one exception, altered immunoelectrophoretic mobility of IgA1 . Only limited removal of other carbohydrates was observed with poor correlation to exoglycosidase activities measured with chromogenic substrates . Desialylation increases the susceptibility of glycoproteins, including IgA1, to proteolysis . Therefore, the desialylation of IgA1 by oral gram-positive rods may facilitate the proteolytic activities of other oral bacteria, and the concerted action may positively influence the survival of the bacteria in the oral community.

Klin Monatsbl Augenheilkd, 1994 Dec, 205(6), 348 - 57
{Normal flora of the human conjunctiva: examination of 135 persons of various ages}; Thiel HJ et al.; PURPOSE OF THE STUDY: Conjunctival flora is a prime suspect in searching for the cause of post-traumatic or post-operative ocular infections . Better information on its individual composition is therefore desirable . METHODS: 135 persons (66 women, 69 men) were examined during a period of 1 1/2 years . The youngest patient was 3 years old, the oldest 90 . Both eyes of all patients were examined . RESULTS: Megasphaera elsdenii, Bacteroides ureolyticus, Bacteroides pneumosintes, Stomatococcus mucilaginosus, and group ANF Corynebacterium were isolated for the first time in the eye by this study . The latter was found so often that it can be ascribed the role of an aerobic indicator organism . Characteristic changes appear in the conjunctival flora at different stages of life . In particular, for example, the species spectrum of aerobic bacteria broadens with advancing age, and the population of aerobic bacteria increases in density . CONCLUSIONS: With increasing age aerobic cocci are found less frequently in favor of the Corynebacteria . The proportion of anaerobic cocci increases among the anaerobes, while Propionibacteria are demonstrated less frequently . The significance of the organisms found in conjunctiva by this study for the first time is still unclear . No qualitative or quantitative sex-specific influence on the development of conjunctival flora was found.

J Antimicrob Chemother, 1994 Dec, 34(6), 965 - 74
Intraphagocytic killing of gram-positive bacteria by ciprofloxacin; Peman J et al.; The intraphagocytic killing of Staphylococcus aureus, Streptococcus pyogenes, and Corynebacterium group D2 by ciprofloxacin (0.1, 1 and 5 mg/L) within human neutrophils was determined . The organisms showed different susceptibility to neutrophil killing mechanisms . The neutrophils with intact and impaired (by phenylbutazone treatment) O2-dependent killing mechanisms were studied . The minimum concentrations of ciprofloxacin to kill 90% of phagocytosed bacteria within untreated neutrophils after 2 h were 1 mg/L for S . aureus and Corynebacterium group D2, and 0.1 mg/L for S . pyogenes . In contrast, exposure for 3 h was required to achieve similar cidal effects within phenylbutazone treated neutrophils . Synergic interaction between ciprofloxacin and the O2-dependent mechanisms of phagocytes was found . The reactive oxygen metabolites produced in the respiratory burst did not affect the intraphagocytic activity of ciprofloxacin . Phenylbutazone treatment of phagocytes would be a good experimental model to study intraphagocytic killing by drugs in situations where the oxidative mechanisms of neutrophils are impaired (for example AIDS and chronic granulomatous disease).

J Antimicrob Chemother, 1994 Dec, 34(6), 1037 - 40
The in-vitro susceptibilities of toxigenic strains of Corynebacterium diphtheriae isolated in northwestern Russia and surrounding areas to ten antibiotics; Maple PA et al.; The in-vitro activities of ten antibiotics against 83 toxigenic strains of Corynebacterium diphtheriae recently isolated in northwestern Russia and surrounding areas were determined by an agar dilution method . All of the strains were susceptible to erythromycin, penicillin, ampicillin, cefuroxime, chloramphenicol, ciprofloxacin, gentamicin and tetracycline . Trimethoprim and rifampicin were each active against 81 isolates, the two strains resistant to the latter agent having been isolated from two members of the same family.

Ther Immunol, 1994 Dec, 1(6), 319 - 24
Induced regression of bovine papillomas by intralesional immunotherapy; Hall H et al.; It has long been assumed that papilloma regression is mediated by immunological mechanisms which are probably cellular in nature . The potentiation of these responses may alter the course of papilloma progression . Certain strains of the bacterium Corynebacterium parvum (Propionibacterium acnes) have been shown to augment cellular immune mechanisms by increasing both macrophage and natural killer cell activity . This study involves the use of naturally occurring bovine papillomas to investigate the immune mechanisms involved in induced papilloma regression . Papillomas were treated by intralesional injection of a C . parvum suspension . Treated papillomas were biopsied at various stages of regression . Tissue samples were subjected to immunohistochemical staining to identify specific infiltrating cells . Results showed that intralesional administration of C . parvum was capable of inducing regression of bovine papillomas in 8-15 weeks . Immunological staining revealed that regression was associated with an increased number of CD8+ and gamma delta+ cells in the dermis, as well as a marked infiltration of neutrophils.

Biochim Biophys Acta, 1994 Nov 23, 1196(1), 14 - 20
Mechanism of alanine excretion in recombinant strains of Zymomonas mobilis; Ruhrmann J et al.; A thiamine-auxotrophic strain of Zymomonas mobilis (CP4thi/pZY73), in which the alaD gene of Bacillus sphaericus coding for the alanine dehydrogenase was expressed, synthesizes and excretes alanine at high rates after thiamine starvation and in the presence of high external ammonium concentrations . The mechanism of alanine excretion was studied in this recombinant Zymomonas mobilis strain . Under production conditions the internal alanine concentration reached values of up to 280 mM and excretion rates of up to 140 nmol min-1 mg dry mass-1 were obtained . The membrane integrity and the energetic properties of the cells remained intact and were comparable to growing wild-type cells . Unspecific leakage of solutes was not observed . We did not find any indication of a carrier-mediated excretion of alanine, since typical properties of this type of mechanism, i.e., saturation at increasing internal substrate concentration, substrate specificity and functional inhibition were absent . Furthermore, a counterflow maximum, which would indicate the involvement of a carrier protein, was not observed either . Consequently, alanine excretion in recombinant Z . mobilis cells is interpreted as mediated by simple diffusion through the intact cytoplasmic membrane at high rates (diffusion constant 10(-8) l s-1 mg dry mass-1 or 0.28 min-1) . For comparison, the diffusion constant for alanine efflux was also measured in Corynebacterium glutamicum cells and the values obtained were significantly lower than those determined in Z . mobilis . The consequences of this finding are discussed.

FEMS Microbiol Lett, 1994 Nov 1, 123(3), 343 - 7
Corynebacterium glutamicum DNA is subjected to methylation-restriction in Escherichia coli; Tauch A et al.; Efficient electroporation of Escherichia coli with plasmid DNA isolated from Corynebacterium glutamicum depends on the use of Mcr-deficient E . coli strains . The transformation frequency increased nearly 800-fold when the Mcr-deficient E . coli DH5 alpha MCR was used instead of E . coli DH5 alpha . We used E . coli strains with different mutations in the methyl-specific restriction systems to show that McrBC-deficiency is sufficient to generate this effect . The results imply that C . glutamicum DNA contains methylcytosine in specific sequences recognized by the E . coli McrBC system.

Toxicol Appl Pharmacol, 1994 Nov, 129(1), 36 - 45
Kupffer cell stimulation with Corynebacterium parvum reduces some cytochrome P450-dependent activities and diminishes acetaminophen and carbon tetrachloride-induced liver injury in the rat; Raiford DS et al.; Chemical activation of Kupffer cells in vivo by vitamin A or latex beads is associated with a worsening of hepatic injury induced by the P450-dependent hepatotoxins acetaminophen (ACET) and carbon tetrachloride (CCl4) and by the P450-independent toxin galactosamine (GLN) . Immunostimulants such as Corynebacterium parvum (CP) also activate Kupffer cells, but do so while prompting release of soluble mediators which depress microsomal oxidative activities in cultured hepatocytes . Therefore, we sought to characterize the effects of CP on hepatic injury in vivo due to ACET and CCl4 while employing GLN as a control . Hepatic microsomal oxidative activity and glutathione (GSH) disposition were examined since each influences susceptibility to injury from ACET or CCl4 . Rats were given CP 28 mg/kg i.v . 5 days before challenge with hepatotoxicant . Hepatic injury was assessed 24 hr after hepatotoxicant administration by measurement of serum alanine aminotransferase (ALT) activity and review of histological sections . Livers from parallel groups of rats were used to prepare microsomal and cytosolic fractions, to measure tissue GSH, or for perfusion to assess GSH efflux . Significant reductions in injury due to ACET or CCl4 were observed while injury due to GLN was potentiated . Serum ALT levels after ACET were 3000 +/- 620 in controls vs 170 +/- 45 IU/liter in the CP-treated group and ALT levels after CCl4 were 3100 +/- 500 in controls vs 1700 + 450 IU/liter in the CP-treated group . In contrast, serum ALT levels after GLN were 920 +/- 230 in controls vs 1700 +/- 370 in the CP-treated group . Patterns of hepatic injury observed on histological sections were those characteristic for each toxin and the severity of injury correlated well with alterations in serum ALT levels for each agent . Hepatic microsomal fractions from rats pretreated with CP showed significantly diminished total cytochrome P450 content as well as reduced activity for two P450IIE1 substrates, p-nitrophenol and 7-ethoxycoumarin . While sinusoidal efflux of GSH increased by 40% in rats pretreated with CP and cytosolic glutathione-S-transferase activity fell slightly, tissue GSH levels were unaffected . These data demonstrate that CP decreases microsomal cytochrome P450 content, reduces biotransformation of two P450IIE1 substrates, and diminishes ACET- and CCl4-induced hepatic injury . In contrast, hepatic injury due to the P450-independent toxin GLN was enhanced . Thus, chemical and immune stimulation of Kupffer cells may result in divergent effects on susceptibility to injury from individual hepatotoxins.

J Leukoc Biol, 1994 Nov, 56(5), 666 - 70
Cellular composition of Corynebacterium pseudotuberculosis pyogranulomas in sheep; Pepin M et al.; The cellular composition of 46 pyogranulomas experimentally induced with Corynebacterium pseudotuberculosis in sheep was determined by immunohistochemistry . Lesions localized in inoculation sites or draining lymph nodes consisted of macrophage and lymphocyte layers distributed around a necrotic center and surrounded by a fibrous capsule . In immature lesions, T cells of the CD4+ subset predominated, whereas in mature lesions proportions of CD8+ T lymphocytes and cells expressing the gamma/delta chains for the T cell receptor increased . Numerous pyogranuloma cells expressed the interleukin-2 receptor . In addition to these general characteristics, a large individual variability in the proportions of macrophage and T cell subsets was observed for lesions of the same age, in particular for epithelioid macrophages . This heterogeneity suggests a different cellular pattern in relation to the persistence or the elimination of bacteria by the host.

J Bacteriol, 1994 Nov, 176(22), 6892 - 9
Quantitative discrimination of carrier-mediated excretion of isoleucine from uptake and diffusion in Corynebacterium glutamicum; Zittrich S et al.; The efflux of isoleucine in whole cells of Corynebacterium glutamicum was studied . The different amino acid fluxes across the plasma membrane were functionally discriminated into passive diffusion, carrier-mediated excretion, and carrier-mediated uptake . Detailed kinetic analysis was made possible by controlled variation of internal isoleucine from low concentrations to 100 mM by feeding with mixtures of isoleucine-containing peptides . Isoleucine diffusion was experimentally separated and proceeded with a first-order rate constant of 0.083 min-1 or 0.13 microliters.min-1.mg (dry mass)-1, which corresponds to a permeability of 2 x 10(-8) cm.s-1 . Uptake of isoleucine was constant at a rate of 1.1 nmol.min-1.mg (dry mass)-1 . Carrier-mediated isoleucine excretion was zero below a threshold of 8 mM cytosolic isoleucine . Above this level, a Michaelis-Menten-type kinetics was observed, with a Km of 21 mM (13 mM plus 8 mM threshold value) and a Vmax of 14.5 nmol.min-1.mg (dry mass)-1 . The activity of the isoleucine excretion carrier depended on the presence of a membrane potential . Excretion was specific for L-isoleucine (and presumably L-leucine) and could be inhibited by SH reagents.

Clin Infect Dis, 1994 Nov, 19(5), 897 - 901
Native-valve endocarditis due to CDC coryneform group ANF-3: report of a case and review of corynebacterial endocarditis; Petit PL et al.; Endocarditis due to a bacterium of CDC coryneform group ANF-3 developed in the native aortic valve of a patient without predisposing factors other than valvular calcification . A review of the literature indicates that corynebacteria are an increasingly important cause of endocarditis . Problems in identification and treatment remain . Techniques for the culture and quick identification of these organisms and effective regimens for treatment of the infections they cause are needed.

Mol Microbiol, 1994 Nov, 14(3), 571 - 81
Identification of IS1206, a Corynebacterium glutamicum IS3-related insertion sequence and phylogenetic analysis; Bonamy C et al.; Integration of plasmid pCGL320 into a Corynebacterium glutamicum ATCC21086 derivative led to tandem amplification of the inserted plasmid (Labarre et al., 1993) . One amplification event was associated with integration of an insertion sequence that we have named IS1206 . Hybridizing sequences were only found in C . glutamicum strains and at various copy numbers . IS1206 is 1290 bp long, carries 32 bp imperfect inverted repeats and generates a 3 bp duplication of the target DNA upon insertion . IS1206 presents the features characteristic of the IS3 family and part of the DNA sequence centering on the putative transposase region (orfB) is similar to those of IS3 and some other related elements . Phylogenetic analysis of orfB deduced protein sequences from IS1206 and IS3-related elements contradicts the phylogeny of the species, suggesting that evolution of these elements might be complex . Horizontal transfer could be invoked but other alternatives like ancestral polymorphism or/and different rates of evolution could also be involved.

What Is Bioassay?, What Is Bioreactor?, What Is Staphylococcus Aureus?, What Is Botulism?, What Is Bioengineering?, c, Bacteria, s, Bacterium, r, Microbes, e, Bacteriology, a, Microbe, s, Pseudomonas aeruginosa, r, Bioreactor, a, Microorganism, s, Erwinia, i, Typhus, e, Listeriosis, r, Lactococci, c, Klebsiella, c, Mueller hinton broth, o, Escherichia coli, c, Bacillus, n, Microorganism, e, Staphylococcus, e, S. cerevisiae, r, Antibiotics, c, Cell suspensions, a, Halophilic bacterium, c, Minimal inhibiting concentration, r, Yeasts, a, Staphylococcus aureus, n, Salmonella typhimurium

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005