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