Int J Syst Bacteriol, 1998 Apr, 48 Pt 2, 489 - 94 Corynebacterium thomssenii sp . nov., a Corynebacterium with N-acetyl-beta-glucosaminidase activity from human clinical specimens; Zimmermann O et al.; A strain of a previously undescribed non-lipophilic coryneform bacterium was isolated from pleural fluids of a patient with chronic renal failure, stroke and pneumonia . Slow fermentative acid production from glucose, maltose and sucrose, and strong N-acetyl-beta-glucosaminidase activity were the most characteristic features of the bacterium . Chemotaxonomic characterization unambiguously indicated that the organism belonged to the genus Corynebacterium . The results of comparative 16S rRNA gene sequence analysis revealed that the isolate represented a new species within the genus, for which the name Corynebacterium thomssenii sp . nov . is proposed . The type strain is DSM 44276. Int J Syst Bacteriol, 1998 Apr, 48 Pt 2, 463 - 8 Corynebacterium camporealensis sp . nov., associated with subclinical mastitis in sheep; Fernandez-Garayzabal JF et al.; Four strains of a hitherto-unknown catalase-positive, facultatively anaerobic Corynebacterium species were isolated from the milk of sheep affected by subclinical mastitis . The most characteristic phenotypic reactions of the four strains were their weak fermentative acid production from glucose, their failure to produce acid from mannitol, xylose, sucrose and maltose, and a strong CAMP reaction with Staphylococcus aureus . Chemotaxonomic investigations revealed the presence of a cell wall based on meso-diaminopimelic acid and short-chain mycolic acids, which is consistent with the genus Corynebacterium . A comparative 16S rRNA gene sequence analysis confirmed that the organisms from sheep were members of the genus Corynebacterium, where they formed a distinct subline, exhibiting > 4% sequence divergence with other known Corynebacterium species . Based on both phenotypic and phylogenetic findings, a new species, Corynebacterium camporealensis, is proposed . The type strain of Corynebacterium camporealensis is CECT 4897 (= CRS-51T). Mol Cell Probes, 1998 Aug, 12(4), 191 - 9 Analysis of the 16S rRNA gene sequence of the coryneform bacterium associated with hyperkeratotic dermatitis of athymic nude mice and development of a PCR-based detection assay; Duga S et al.; By 16S rDNA sequencing the authors have characterized the coryneform bacteria associated with hyperkeratotic dermatitis (HD) of athymic nude mice isolated from six different outbreaks of the disease in Northern Italy . This analysis has allowed the authors to confirm the classification of the bacteria as Corynebacterium bovis and to develop a 16S rDNA-based polymerase chain reaction (PCR) detection assay . The test was performed directly on the DNA extracted from epidermal swabs . The PCR primers were chosen to match the 16S rDNA sequence fragments which differ most from the other Corynebacterium spp . The test was shown to be both sensitive and specific for C . bovis . Detection of as few as three viable bacterial cells was possible with the use of an oligonucleotide probe in a liquid hybridization assay. J Bacteriol, 1998 Sep, 180(17), 4650 - 7 Intraspecific variation of unusual phospholipids from Corynebacterium spp . containing a novel fatty acid; Niepel T et al.; The novel fatty acid trans-9-methyl-10-octadecenoic acid was isolated from the coryneform bacterial strain LMG 3820 (previously misidentified as Arthrobacter globiformis) and identified by spectroscopic methods and chemical derivatization . This fatty acid is attached to the unusual lipid acyl phosphatidylglycerol . Five different species of this lipid type were identified; their structures were elucidated by tandem mass spectrometry and are reported here for the first time . Additionally, we identified three different cardiolipins, two bearing the novel fatty acid . The characteristic 10-methyl-octadecanoic acid was present only in phosphatidylinositol . Because of the unusual fatty acid pattern of strain LMG 3820, the 16S rDNA sequence was determined and showed regions of identity to sequences of Corynebacterium variabilis DSM 20132(T) and DSM 20536 . All three strains possessed the novel fatty acid, identifying trans-9-methyl-10-octadecenoic acid as a potential biomarker characteristic for this taxon . Surprisingly, the fatty acid and relative abundances of phospholipids of Corynebacterium sp . strain LMG 3820 were similar to those of the type strain but different from those of Corynebacterium variabilis DSM 20536, although all three strains possessed identical 16S rDNA sequences and strains DSM 20132(T) and DSM 20536 have 90.5% DNA-DNA homology . This is one of the rare cases wherein different organisms with identical 16S rDNA sequences have been observed to present recognizably different fatty acid and lipid compositions . Since methylation of a fatty acid considerably lowers the transition temperature of the corresponding lipid resulting in a more flexible cell membrane, the intraspecific variation in the lipid composition, coinciding with the morphological and Gram stain reaction variability of this species, probably offers an advantage for this species to inhabit different environmental niches. Appl Microbiol Biotechnol, 1998 Jul, 50(1), 42 - 7 Analysis of the leuB gene from Corynebacterium glutamicum; Patek M et al.; The leuB gene of Corynebacterium glutamicum was found to be present on a 2.2-kb BamHI-SacI chromosomal fragment which complemented the leuB mutation of Escherichia coli . The activity of 3-isopropylmalate dehydrogenase (EC 1.1.1.85), encoded by the leuB gene, was significantly increased in C . glutamicum cells harbouring a plasmid containing the 2.2-kb fragment . The nucleotide sequence of the C . glutamicum leuB coding region (an open reading frame, ORF, of 1020 bp encoding a polypeptide of 340 amino acids with M(r) of 36 144) was determined . The deduced amino acid sequence of the product of this ORF is highly homologous to those of 3-isopropylmalate dehydrogenases from three species of mycobacteria . The transcriptional start site of the leuB gene was localized 35 bp upstream of its translational start; a functional terminator was detected in the 3' flanking region . Northern hybridization analysis showed that the C . glutamicum leuB gene is transcribed as a single monocistronic RNA (approximately 1.2 kb in size) . Activity of the leuB promoter was significantly reduced when leucine was present in the growth medium . This suggests the negative regulation of the leuB expression on the transcriptional level in C . glutamicum cells. Bacteriol Virusol Parazitol Epidemiol, 1998 Jan-Jun, 43(1-2), 47 - 51 {Corynebacterium urealyticum in urinary infections in children}; Coman G et al.; The urinary infection caused by C . urealyticum is a rare circumstance, especially in children . Authors present the etiology of these infections, in children admitted in the Pediatric Clinical Hospital "Sf . Maria", Iasi, during 1997 . This recently recognised uropathogen was isolated from three cases . Compared to other etiological agents, the frequency was 0.75% . This study presents the diagnosis and identification criteria for C . urealyticum and therapeutic challenges of this infection. Lab Anim, 1998 Jul, 32(3), 330 - 6 Hyperkeratosis-associated coryneform infection in severe combined immunodeficient mice; Scanziani E et al.; Hyperkeratosis-associated coryneform (HAC) is a coryneform bacterium, with a biochemical profile similar to Corynebacterium bovis, that causes hyperkeratotic dermatitis in athymic nude mice . In the present study 28 severe combined immunodeficient (SCID) mice coming from six different animal facilities were submitted for bacteriological and pathological examination . HAC was isolated from 10 SCID mice belonging to two of these facilities . Two of the HAC-infected mice showed macroscopical lesions consisting in large alopecic areas, with small white flakes, involving the dorsum, flanks, neck and cheeks . Histologically, the skin of these animals was characterized by diffuse acanthosis and hyperkeratosis . In the other eight HAC-infected SCID mice no macroscopical lesions were observed but focal areas of minimal to mild acanthosis were histologically detected in five cases . These results suggest that HAC can infect SCID mice inducing skin lesions similar, although generally less severe, to those observed in nude mice with hyperkeratotic dermatitis . Our results pointed out that SCID mice may play an important role in the epidemiology of hyperkeratotic dermatitis of athymic nude mice. Scand J Immunol, 1998 Aug, 48(2), 183 - 91 Hepatoprotective effect of propagermanium on Corynebacterium parvum and lipopolysaccharide-induced liver injury in mice; Yokochi S et al.; Propagermanium is an organic germanium compound with immunopotentiating activity . We examined the hepatoprotective effect of propagermanium and its mechanism in an experimental animal model of acute liver injury induced with Corynebacterium parvum (C . parvum) and lipopolysaccharide (LPS) injection . Oral pretreatment with propagermanium decreased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity in a dose-dependent manner . Significant attenuation of ALT and AST activity was obtained at a dose of 3 mg/kg . Administration of propagermanium also inhibited the infiltration of mononuclear cells into the liver of mice induced by C . parvum/LPS . Immunohistochemical examination revealed infiltration of the liver by CD4-, CD8-, CD11b- and Gr-1-positive cells . Propagermanium prevented CD4- and CD11b-positive cells from infiltrating the liver . In this animal model, blood cytokine levels increased rapidly after LPS injection, causing severe hepatitis . Notably, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) are important mediators of the progress of liver injury . We demonstrated that propagermanium reduced IFN-gamma production by 53% at a dose of 3 mg/kg and also significantly inhibited the production of interleukin-12 (IL-12) . These results indicate that propagermanium inhibits cell infiltration in the liver and cytokine production, and improves massive liver injury in C . parvum/LPS mice. J Biol Chem, 1998 Aug 28, 273(35), 22420 - 7 Motion of the DNA-binding domain with respect to the core of the diphtheria toxin repressor (DtxR) revealed in the crystal structures of apo- and holo-DtxR; Pohl E et al.; The diphtheria toxin repressor (DtxR) from Corynebacterium diphtheriae is a divalent metal-activated repressor of chromosomal genes that encode proteins responsible for siderophore-mediated iron uptake and also of the gene of certain corynebacteriophages that encodes diphtheria toxin . DtxR consists of two 25.3-kDa three-domain subunits and is a member of a family of related repressor proteins in several Gram-positive bacterial species, some of which are important human pathogens . In this paper, we report on the first high resolution crystal structures of apo-DtxR in two related space groups . In addition, crystal structures of Zn-DtxR were determined in the same two space groups . The resolutions of the structures range from 2.2 to 2.4 A . The four refined models of the apo- and the holo-repressor exhibit quite similar metal binding centers, which do, however, show higher thermal motion in the apo-structures . All four structures reported differ from each other in one important aspect . The N-terminal DNA-binding domain and the last 20 residues of the dimerization domain of each subunit move significantly with respect to the core of the DtxR dimer, which consists of residues 74-120 from both subunits . These results provide the first indication of a conformational change that may occur upon binding of the holo-repressor to DNA. Infect Immun, 1998 Sep, 66(9), 4123 - 9 SirR, a novel iron-dependent repressor in Staphylococcus epidermidis; Hill PJ et al.; In Staphylococcus epidermidis and Staphylococcus aureus, a number of cell wall- and cytoplasmic membrane-associated lipoproteins are induced in response to iron starvation . To gain insights into the molecular basis of iron-dependent gene regulation in the staphylococci, we sequenced the DNA upstream of the 3-kb S . epidermidis sitABC operon, which Northern blot analysis indicates is transcriptionally regulated by the growth medium iron content . We identified two DNA sequences which are homologous to elements of the Corynebacterium diphtheriae DtxR regulon, which controls, in response to iron stress, for example, production of diphtheria toxin, siderophore, and a heme oxygenase . Upstream of the sitABC operon and divergently transcribed lies a 645-bp open reading frame (ORF), which codes for a polypeptide of approximately 25 kDa with homology to the DtxR family of metal-dependent repressor proteins . This ORF has been designated SirR (staphylococcal iron regulator repressor) . Within the sitABC promoter/operator region, we also located a region of dyad symmetry overlapping the transcriptional start of sitABC which shows high homology to the DtxR operator consensus sequence, suggesting that this region, termed the Sir box, is the SirR-binding site . The SirR protein was overexpressed, purified, and used in DNA mobility shift assays; SirR retarded the migration of a synthetic oligonucleotide based on the Sir box in a metal (Fe2+ or Mn2+)-dependent manner, providing confirmatory evidence that this motif is the SirR-binding site . Furthermore, Southern blot analysis of staphylococcal chromosomal DNA with the synthetic Sir box as a probe confirmed that there are at least five Sir boxes in the S . epidermidis genome and at least three in the genome of S . aureus, suggesting that SirR controls the expression of multiple target genes . Using a monospecific polyclonal antibody raised against SirR to probe Western blots of whole-cell lysates of S . aureus, S . carnosus, S . epidermidis, S . hominis, S . cohnii, S . lugdunensis, and S . haemolyticus, we identified an approximately 25-kDa cross-reactive protein in each of the staphylococcal species examined . Taken together, these data suggest that SirR functions as a divalent metal cation-dependent transcriptional repressor which is widespread among the staphylococci. Ophthalmology, 1998 Aug, 105(8), 1471 - 7 The effect of successful contact lens wear on mucosal immunity of the eye; McClellan KA et al.; OBJECTIVE: This study aimed to assess the effect of contact lens wear on the mucosal defenses of the outer eye against infection . DESIGN: A case-controlled study of daily contact lens wearers in their initial 6 months of contact lens wear . PARTICIPANTS: Contact lens wearers (mean age, 23.1 years; 47 subjects) were compared with age-matched control subjects (mean age, 24.7 years; 44 subjects) . INTERVENTION: Outer eye defenses were studied by assay of tear constituents and quantitative conjunctival microbiology . MAIN OUTCOME MEASURES: Antimicrobial activity of tears was studied by assay of total immunoglobulin A (IgA), IgA isotype-specific antibodies reactive with Escherichia coli, Haemophilus influenzae, Staphylococcus epidermidis, albumin and lysozyme, and the ocular surface microbial load determined using quantitative microbiology of the conjunctival sac . RESULTS: The IgA isotype-specific antibodies reactive with E . coli (P = 0.03) and S . epidermidis (P = 0.068) were lower in contact lens wearers, but antibody:albumin ratios were not significantly different in the two groups . Contact lens wear also had no significant effect on tear IgA, albumin, or lysozyme or its ratios with albumin . Bacterial numbers and colonization rates for coagulase-negative staphylococci were greater in contact lens wearers than in age-matched control subjects . Corynebacterium sp . and non-Enterobacteriaceae (P = 0.007) were isolated more frequently and in greater numbers from contact lens wearers . Colonization rates were increased for Corynebacterium sp., but non-Enterobacteriaceae were transient . In both daily contact lens wearers and age-matched control subjects, most conjunctival flora were transient rather than colonizing, and no subject developed an outer eye infection during the study . CONCLUSION: These results suggest that daily contact lens wear does not significantly alter the mucosal defenses of the outer eye that function to eliminate organisms from the conjunctival sac and prevent outer eye infection. Nitric Oxide, 1997 Jun, 1(3), 254 - 62 Nitric oxide-induced expression of C-reactive protein in islet cells as a very early marker for islet stress in the rat pancreas; Fehsel K et al.; In searches for marker molecules specifically expressed in nitric oxide-treated islet cells as a means to recognize early events in islet destruction, we now establish the presence of neo-C-reactive protein (neoCRP) in rat islet cells as early as 2 hr after treatment . We detected this altered molecular form of the acute-phase-reactant C-reactive protein (CRP) using immunocytochemistry with an anti-neoCRP-specific monoclonal antibody as well as reverse transcription-polymerase chain reaction with CRP-specific primers and in situ hybridization to demonstrate the presence of CRP-specific mRNA . After induction of a generalized inflammatory reaction in rats with heat-inactivated Corynebacterium parvum in vivo, neoCRP expression in islets is also found and within the pancreas restricted to pancreatic islet cells only . Our findings suggest an early heat-shock-like expression of this molecule in response to local nitrite oxide production or to exogeneously added nitric oxide in islet cells. Mikrobiologiia, 1998 May-Jun, 67(3), 338 - 44 {Electrical response of inner membrane structures of corynebacteria during electrotransformation}; Tiurin MV et al.; The efficiency of the electrotransformation of intact cells of corynebacteria by a solitary impulse with a complex shape amounted to 10(6) transformants/microgram of plasmid pNV1 DNA at an electric field strength of 14.2 kW/cm; the voltage-current curve of the cell samples was nonlinear . Under these conditions, the structure of the electric current impulse passing intact cells or protoplasts included oscillations characterized by increasing amplitude and a duration of 170 microseconds, which were not detected in the structure of the electric current impulses at field strengths insufficient for obtaining transformants . These changes in the impulse shape suggest the involvement of internal closed membrane structures in the electrical response of cells to the exogenous electric impulse . Most probably, under conditions of electrical treatment optimal for transformation, electropores are formed in the intracellular membranes of corynebacteria. Mol Microbiol, 1998 Jul, 29(1), 139 - 50 The cell wall porin of Nocardia farcinica: biochemical identification of the channel-forming protein and biophysical characterization of the channel properties; Riess FG et al.; A channel-forming protein was identified in cell wall extracts of the Gram-positive, strictly aerobic bacterium Nocardia farcinica . The cell wall porin was purified to homogeneity and had an apparent molecular mass of about 87 kDa on tricine-containing SDS-PAGE . When the 87 kDa protein was boiled for a longer time in sodium dodecylsulphate (SDS) it dissociated into two subunits with molecular masses of about 19 and 23 kDa . The 87 kDa form of the protein was able to increase the specific conductance of artificial lipid bilayer membranes from phosphatidylcholine (PC) phosphatidylserine (PS) mixtures by the formation of ion-permeable channels . The channels had on average a single-channel conductance of 3.0 nS in 1M KCl, 10mM Tris-HCl, pH8, and were found to be cation selective . Asymmetric addition of the cell wall porin to lipid bilayer membranes resulted in an asymmetric voltage dependence . The single-channel conductance was only moderately dependent on the bulk aqueous KCl concentration, which indicated point charge effects on the channel properties . The analysis of the single-channel conductance data in different salt solutions using the Renkin correction factor, and the effect of negative charges on channel conductance suggested that the diameter of the cell wall porin is about 1.4-1.6nm . Channel-forming properties of the cell wall porin of N . farcinica were compared with those of mycobacteria and corynebacteria . The cell wall porins of these members of the order Actinomycetales share common features because they form large and water-filled channels that contain negative point charges. Nature, 1998 Jul 30, 394(6692), 502 - 6 Structure of the metal-ion-activated diphtheria toxin repressor/tox operator complex; White A et al.; The virulent phenotype of the pathogenic bacterium Corynebacterium diphtheriae is conferred by diphtheria toxin, whose expression is an adaptive response to low concentrations of iron . The expression of the toxin gene (tox) is regulated by the repressor DtxR, which is activated by transition metal ions . X-ray crystal structures of DtxR with and without (apo-form) its coordinated transition metal ion have established the general architecture of the repressor, identified the location of the metal-binding sites, and revealed a metal-ion-triggered subunit-subunit 'caliper-like' conformational change . Here we report the three-dimensional crystal structure of the complex between a biologically active Ni(II)-bound DtxR(C102D) mutant, in which a cysteine is replaced by an aspartate at residue 102, and a 33-base-pair DNA segment containing the toxin operator toxO . This structure shows that DNA interacts with two dimeric repressor proteins bound to opposite sides of the tox operator . We propose that a metal-ion-induced helix-to-coil structural transition in the amino-terminal region of the protein is partly responsible for the unique mode of repressor activation by transition metal ions. Microbiology, 1998 Jul, 144 ( Pt 7), 1863 - 8 A novel system with positive selection for the chromosomal integration of replicative plasmid DNA in Corynebacterium glutamicum; Ikeda M et al.; A simple system has been developed for generating Corynebacterium glutamicum strains containing stable replicative plasmids integrated into the chromosome via homologous recombination . The system is based upon extremely strong incompatibility between two plasmids, which cannot be co-maintained even under antibiotic selective pressure . Integration of the resident plasmid that contained the trpD gene of C . glutamicum was achieved by introduction of a second plasmid and subsequent selection for the maintenance of both plasmids . Plasmid integrates positive for both plasmid markers were obtained at a frequency about 10(-3) of the normal transformation frequency with selection for the maintenance of only the second plasmid . Southern analysis revealed that the integration had occurred through a single-crossover homologous recombination between the trpD regions of the host genome and the plasmid . On the basis of the Campbell-type integration, chromosome walking was attempted by using Escherichia coli replication origins that were also present in the integrated plasmid . The chromosomal DNA was digested, ligated, and used to transform E . coli, which enabled recovery of the expected adjacent genomic DNA regions . The plasmid integrate was stably maintained for 30 generations under non-selective culture conditions, suggesting that the integrated sequences carrying a replicon active in the host were maintained as a stable chromosomal insert in C . glutamicum. Microbiology, 1998 Jul, 144 ( Pt 7), 1853 - 62 The role of the Corynebacterium glutamicum rel gene in (p)ppGpp metabolism; Wehmeier L et al.; To investigate the metabolism of (p)ppGpp in amino-acid-producing coryneform bacteria, a PCR-based strategy using degenerate consensus oligonucleotides was applied to isolate the rel gene of Corynebacterium glutamicum ATCC 13032 . The gene consists of 2283 nucleotides and encodes a protein of 760 amino acids with a molecular mass of 84.4 kDa . The amino acid sequence revealed extensive similarities to the related proteins RelA and SpoT of Escherichia coli, which are known to be involved in (p)ppGpp biosynthesis and degradation . The C . glutamicum rel gene is located downstream of the apt gene encoding an adenine phosphoribosyltransferase, and an ORF with similarities to dciAE, which represents part of a dipeptide transport system in E . coli . A C . glutamicum mutant strain carrying a defined deletion in the rel gene was constructed . This mutant failed to accumulate (p)ppGpp in response to amino acid starvation . When overexpressed in E . coli, the C . glutamicum rel gene was able to reverse growth defects caused by an overexpressed relA gene . It is proposed that the C . glutamicum rel gene encodes a bifunctional enzyme with (p)ppGpp synthetase and (p)ppGpp-degrading activities. Vet Microbiol, 1998 May, 62(2), 135 - 43 Comparative molecular characterization of Corynebacterium pseudotuberculosis of different origin; Costa LR et al.; Ribotyping and susceptibility to 17 antimicrobial agents were used to compare 37 isolates of Corynebacterium pseudotuberculosis (28 from horses, 1 from cattle, 3 from sheep and 5 from goats) derived from various types of lesions, and different geographic locations . According to the presence of nitrate reductase, all but one isolate from horses reduced nitrate (nitrate-positive), whereas all isolates from sheep and goats were unable to reduce nitrate (nitrate-negative) . The ribotype of the nitrate-negative isolate from a horse with ulcerative lymphangitis was identical to all the other isolates from horses, and different than the ribotype of nitrate-negative isolates from sheep and goats . Ribotyping with one of the restriction endonucleases, Apa 1, revealed differences between, but not within, the two biotypes . However, ribotyping with Pst 1 endonuclease revealed one variant within the equine biotype and one variant within the ovine biotype . The minimum inhibitory concentration (MIC; microgram/ml) of antimicrobial agents against isolates from nitrate-negative and nitrate-positive groups was very similar, with the exception of isolates from sheep and goats which had a higher MIC for amikacin than isolates from horses and cattle. Biosci Biotechnol Biochem, 1998 Jun, 62(6), 1055 - 60 Action of poly (alpha-L-guluronate)lyase from Corynebacterium sp . ALY-1 strain on saturated oligoguluronates; Matsubara Y et al.; A kinetic analysis of degradation of saturated oligoguluronates by poly(alpha-L-guluronate)lyase from Corynebacterium sp . ALY-1 strain was done . The saturated oligoguluronates were prepared by hydrolyzing poly alpha-1,4-L-guluronate from alginate with HCl, and then by gel filtration on a Bio-Gel P-6 column . The saturated pentaguluronate or above were rapidly degraded by the enzyme, while tetraguluronate was slowly degraded . From the dependency of the catalytic rate constant (kcat) on the degree of polymerization of substrates, the enzyme was found to have a subsite size corresponding to hexaguluronate units . The action pattern of the enzyme on hexaguluronate suggested that the catalytic site of the enzyme was matched to the linkage between the second and third uronic residue from the non-reducing end, since the substrate was mainly split into a unsaturated tetramer and a saturated dimer from a HPLC analysis. Prev Vet Med, 1998 Jun 30, 35(4), 241 - 53 Application and evaluation of a mailed questionnaire for an epidemiologic study of Corynebacterium pseudotuberculosis infection in horses; Doherr MG et al.; The objective of this study is to describe the design, application and validity of a self-administered (mailed) questionnaire to collect data on potential risk factors for Corynebacterium pseudotuberculosis infection in California horses . Horses admitted to the UC Davis Veterinary Medical Teaching Hospital (VMTH) between 1 July 1992 and 30 June 1994 served as the study base for case identification and simple random sampling of 800 control horses . A questionnaire was mailed to owners of the study horses, followed by a reminder postcard and a second copy of a questionnaire . Data were collected on owner and horse identity and demographics, horse management and use, geographic location, and general health-related issues . Return pattern over time as well as differential return proportions were described . The overall return proportion was 66% (587/890), and the completion proportion 55% (491/890) . The number of returns over time followed a negative binomial distribution, with over 90% of all returns being in by the end of the fifth week after mailing, and over 99% at the end of the tenth week . Some categories within the variables age (between 2 and 3 years), breed (Thoroughbred and Standardbred horses) and gender (stallions) had significantly lower return proportions than expected (differential return; p < 0.05) . The profile of these horses fits a section of the racehorse population that is served by the VMTH . Age, breed and disease status information was available from the VMTH medical records and from the questionnaire, and was used to determine the validity of the survey data . There was good agreement between the data from the two sources, and we therefore concluded that the quality of the survey information was sufficient to perform a risk-factor analysis . The mailed survey provided a rapid and cost-effective method of collecting additional information to supplement existing medical records. Prev Vet Med, 1998 Jun 30, 35(4), 229 - 39 Risk factors associated with Corynebacterium pseudotuberculosis infection in California horses; Doherr MG et al.; A case-control study was designed using equine medical records from the UC Davis Veterinary Medical Teaching Hospital (VMTH) and data derived through a mailed survey . The objective was to evaluate the associations between horse demographics, horse-management factors, and equine Corynebacterium pseudotuberculosis infection in California . Horses admitted to the VMTH between July 1 1992 and June 30 1994 served as the study base for case identification and simple random sampling of 800 controls . A questionnaire was mailed to the owners of all horses enrolled in the study to collect data on demographics, management and health-related questions . A logistic-regression model containing age, outdoor activity level, other locations in California, insect-control measures, contact with other horses, and summer pasture was developed . The final model was adjusted for the suspected confounding variables admission type, regular teaching hospital patient and breed . Horses of age between 1 and 2 yrs and between 3 and 5 yrs, and horses in contact with other horses or horses on summer pasture had significantly increased odds (p < 0.05) of being diagnosed with C . pseudotuberculosis infection . The results support the hypotheses that the disease predominantly affects young-adult horses of all breeds and both sexes, and that management factors play an important role in occurrence of the disease . Since the existing serological test system is not reliable and destruction of infected animals is not feasible, the most-logical approach for disease prevention is the early identification and isolation of clinical cases and the implementation of management changes like improvement of stable hygiene and insect control and change of pasture practices. Curr Microbiol, 1998 Sep, 37(3), 156 - 8 Bacteriological properties of a sucrose-fermenting Corynebacterium diphtheriae strain isolated from a case of endocarditis; de Mattos-Guaraldi AL et al.; An atypical sucrose-fermenting Corynebacterium diphtheriae strain was isolated from three blood cultures of a 14-year-old girl presented to a university teaching hospital in Rio de Janeiro, Brazil . She had mitral endocarditis that proved to be fatal despite intensive antibiotic therapy . Blood cultures showed a fluorescent Gram-positive, aerobic, coryneform-like bacillus presenting pyrazinamidase and CAMP reaction negative . The isolate was identified as a toxigenic strain of C . diphtheriae var . mitis by both Elek and radial immunodiffusion (RID) tests . The invasive C . diphtheriae sucrose-fermenting biotype strain adhered to glass surfaces and expressed pronounced hemagglutinating activity (titer 8), a property common among the nonfermenting biotype strains . Laboratories should be alert to the possibility of the isolation of C . diphtheriae with a positive sucrose fermentation test, especially when nontoxigenic strains are isolated from uncommon anatomic sites. Zentralbl Hyg Umweltmed, 1998 Jun, 201(2), 167 - 88 {Danger of infection from communion cups--an underestimated risk?}; Fiedler K et al.; The problem of a risk of infection from the common use of chalices has been discussed controversially in literature . Opinions were mainly based on laboratory experiments and theoretical considerations . The authors examined bacterial counts and species existing under normal conditions after communion . For this purpose, contact samples were taken from the inside and outside of chalices at the rim . Staphylococci and alpha-haemolytic streptococci were found on all chalices examined . On more than 80%, there were apathogenic micrococci, nonhaemolytic streptococci, apathogenic neisseria and apathogenic corynebacteria as well as lactobacilli and bacilli . Staphylococcus aureus was found on 26.4% of chalices . Although the risk of infection for healthy persons from a commonly used chalice can be rated as low, it should not be underestimated for persons with reduced resistance and immunocompetence, or with reduced defences as a result of therapeutic measures . From the hygienic point of view, the most favourable approaches to avoid infection would be the use of individual chalices for all participants in the communion or the immersion of wafers or bread in wine or in grape juice by the priest (intinction). Chemotherapy, 1998 Jul-Aug, 44(4), 230 - 7 Antibiotic susceptibility testing (agar disk diffusion and agar dilution) of clinical isolates of Corynebacterium jeikeium; Traub WH et al.; Thirty-three clinical isolates of Corynebacterium jeikeium were examined for susceptibility to 27 antimicrobial drugs with the agar dilution test . Sheep-blood-supplemented Mueller-Hinton agar performed better than Wilkins-Chalgren agar . Disk susceptibility (Bauer-Kirby) tests were carried out in parallel with 24 of the chemotherapeutic agents . All isolates were susceptible to teicoplanin and vancomycin . All isolates resisted fosfomycin, mupirocin, and trimethoprim-sulfamethoxazole . The isolates varied in susceptibility to ciprofloxacin, doxycycline, fusidic acid, ofloxacin, and tetracycline; most were susceptible to rifampin . Surprisingly few discrepancies between agar dilution and disk diffusion tests were encountered when utilizing NCCLS interpretive criteria currently valid for enterococcal isolates. Proc Natl Acad Sci U S A, 1998 Jul 21, 95(15), 8916 - 21 A bacterial cytokine; Mukamolova GV et al.; Viable cells of Micrococcus luteus secrete a factor, which promotes the resuscitation and growth of dormant, nongrowing cells of the same organism . The resuscitation-promoting factor (Rpf) is a protein, which has been purified to homogeneity . In picomolar concentrations, it increases the viable cell count of dormant M . luteus cultures at least 100-fold and can also stimulate the growth of viable cells . Rpf also stimulates the growth of several other high G+C Gram-positive organisms, including Mycobacterium avium, Mycobacterium bovis (BCG), Mycobacterium kansasii, Mycobacterium smegmatis, and Mycobacterium tuberculosis . Similar genes are widely distributed among high G+C Gram-positive bacteria; genome sequencing has uncovered examples in Mycobacterium leprae and Mb . tuberculosis and others have been detected by hybridization in Mb . smegmatis, Corynebacterium glutamicum, and Streptomyces spp . The mycobacterial gene products may provide different targets for the detection and control of these important pathogens . This report is thus a description of a proteinaceous autocrine or paracrine bacterial growth factor or cytokine. Proc Natl Acad Sci U S A, 1998 Jul 21, 95(15), 8823 - 8 Up-regulation of inducible nitric oxide synthase expression in cancer-prone p53 knockout mice; Ambs S et al.; High concentrations of nitric oxide (NO) cause DNA damage and apoptosis in many cell types . Thus, regulation of NO synthase (NOS) activity is essential for minimizing effects of cytotoxic and genotoxic nitrogen oxide species . We have shown previously that NO-induced p53 protein accumulation down-regulates basal and cytokine-modulated inducible NOS (NOS2) expression in human cells in vitro . To further characterize the feedback loop between NOS2 and p53, we have investigated NO production, i.e., urinary nitrate plus nitrite excretion, and NOS2 expression in homozygous p53 knockout (KO) mice . We report here that untreated p53 KO mice excreted 70% more nitrite plus nitrate than mice with wild-type (wt) p53 . NOS2 protein expression was constitutively detected in the spleen of untreated p53 KO mice, whereas it was undetectable in the spleen of wt p53 controls . Upon treatment with heat-inactivated Corynebacterium parvum, urinary nitrite plus nitrate excretion of p53 KO mice exceeded that of wt controls by approximately 200% . C . parvum treatment also induced p53 accumulation in the liver . Splenectomy reduced the NO output of C . parvum-treated p53 KO mice but not of wt p53 controls . Although NO production and NOS2 protein expression were increased similarly in KO and wt p53 mice 10 days after injection of C . parvum, NOS2 expression returned to baseline levels only in wt p53 controls while remaining up-regulated in p53 KO mice . These genetic and functional data indicate that p53 is an important transrepressor of NOS2 expression in vivo and attenuates excessive NO production in a regulatory negative feedback loop. Mikrobiol Z, 1998 Mar-Apr, 60(2), 60 - 4 {The effect of antibiotics on the adhesion of Corynebacterium diphtheriae to buccal epithelial cells}; Hladka OA et al.; The model of buccal epithelium of healthy people was used to study the effect of antibiotics with different action mechanism (benzylpenicillin, ampicylin, erythromycin, streptomycin) on the adhesion intensity of diphtheria corynebacteria . It has been established that only erythromycin in minimum inhibiting concentration manifested the ability to inhibit the adhesion of different biovars of Corynebacterium diphtheriae . The drug action did not depend on the level of adhesive activity of certain cultures . Subinhibiting concentrations of erythromycin did not affect the processes of the diphtheria corynebacteria adhesion. Mol Cells, 1998 Jun 30, 8(3), 286 - 94 Isolation and analysis of metA, a methionine biosynthetic gene encoding homoserine acetyltransferase in corynebacterium glutamicum; Park SD et al.; The metA gene encoding homoserine acetyltransferase, the first enzyme of the methionine biosynthetic pathway, was isolated from a pMT1-based corynebacterium glutamicum gene library via complementation of an Escherichia coli metA mutant . A DNA-sequence analysis of the cloned DNA is identified an open-reading frame of 1,137 bp which encodes a protein with the molecular weight of 41,380 comprising 379 amino acids . The putative protein product showed good amino acid-sequence homology to its counterpart in other organisms . The internal fragment of the cloned DNA was successfully used to disrupt chromosomal metA, demonstrating the identity of the cloned gene . The C . glutamicum metA mutant lost the ability to grow on glucose minimal medium supplemented with homoserine . However, the mutant could grow on a minimal medium supplemented with cystathionine, demonstrating that C . glutamicum uses the cystathionine route to synthesize methionine . Introduction of a plasmid carrying cloned metA into C . glutamicum resulted in a 10-fold increase in enzyme activities and expression of a protein product of M(r) 41,000, which agrees with the sequence data and is similar in size to those of other homoserine acetyltransferases . Unlike E . coli whose metA product uses succinyl coenzyme A as a substrate, the cloned metA gene produced homoserine acetyltransferase which uses only acetyl coenzyme A as the acyl donor. Eur J Biochem, 1998 Jun 1, 254(2), 395 - 403 Biochemical and genetic characterization of the membrane-associated malate dehydrogenase (acceptor) from Corynebacterium glutamicum; Molenaar D et al.; In addition to a cytoplasmic, NAD-dependent malate dehydrogenase (EC 1.1.1.37), Corynebacterium glutamicum possesses a highly active membrane-associated malate dehydrogenase (acceptor) (EC 1.1.99.16) . This enzyme also takes part in the citric acid cycle . It oxidizes L-malate to oxaloacetate and donates electrons to ubiquinone-1 and other artificial acceptors or, via the electron transfer chain, to oxygen . NAD is not an acceptor and the natural direct acceptor for the enzyme is most likely a quinone . The enzyme is therefore called malate:quinone oxidoreductase, abbreviated to Mqo . Mqo is a peripheral membrane protein and can be released from the membrane by addition of chelators . The solubilized form was partially purified and characterized biochemically . FAD is probably a tightly but non-covalently bound prosthetic group, and the enzyme is activated by lipids . A C . glutamicum mutant completely lacking Mqo activity was isolated . It grows poorly on several substrates tested . The mutant possesses normal levels of cytoplasmic NAD-dependent malate dehydrogenase . A plasmid containing the gene from C . glutamicum coding for Mqo was isolated by complementation of the Mqo-negative phenotype . It leads to overexpression of Mqo activity in the mutant . The nucleotide sequence of the mqo gene was determined and is the first sequence known for this enzyme . The derived protein sequence is similar to hypothetical proteins from Escherichia coli, Klebsiella pneumoniae, and Mycobacterium tuberculosis. Rev Hosp Clin Fac Med Sao Paulo, 1998 Jan-Feb, 53(1), 3 - 5 {Intertrigo in patients with lower limb lymphedema . Clinical and laboratory correlation}; de Andrade MF et al.; Cutaneous lesions in the interdigital spaces are commonly seen in lymphedema patients and their prevention and suitable care is one of the cornerstones of any successful treatment, by preventing acute inflammations and additional worsening in limb volume and fibrosis . We obtained swab specimens from the interdigital area from 21 patients followed in the Lymphedema Unit of the Department of Vascular Surgery of the University of Sao Paulo; thirteen of them had lesions suggestive of tinea pedis . The pathological agent could be identified in 11 out of these 13 patients: fungal infection alone was responsible for seven lesions, Corynebacterium minutissimum for another two and both agents were isolated from two patients . Although two patients had evident clinical lesion of the skin, no fungal or bacterial species could be isolated . From the eight patients without interdigital lesions, Candida and Corynebacterium was found in one . We concluded that clinical examination has a high sensibility (84%) and specificity (91%) but the high prevalence of Corynebacterium minutissimum suggests that adequate treatment should follow careful laboratory examination. Biochemistry, 1998 Jul 7, 37(27), 9716 - 23 Proposed steady-state kinetic mechanism for Corynebacterium ammoniagenes FAD synthetase produced by Escherichia coli; Efimov I et al.; The bifunctional enzyme, FAD synthetase (FS), from Corynebacterium ammoniagenes was overproduced in Escherichia coli and purified, and its steady-state kinetic properties were investigated . Although FMN is an intermediate product in the conversion of riboflavin to FAD, FMN must be released after formation, and then rebind for adenylylation . It was shown that adenylylation of FMN is reversible; FAD and pyrophosphate can be converted to FMN and ATP by the enzyme . In contrast, under the conditions studied, phosphorylation of riboflavin is irreversible . A method is described for analysis of two catalytic cycles, occurring on one enzyme, which have a substrate and/or product in common . The binding order for the phosphorylation cycle of FS was established as riboflavin(in), ATP(in), ADP(out), and FMN(out) . The order for the adenylylation cycle was ATP(in), FMN(in), pyrophosphate(out), and FAD(out) . A set of steady-state constants was determined, and without additional optimization, these constants were sufficient to describe experimental progress curves for conversion of riboflavin to FAD . In independent studies, it was demonstrated that FMN binds to apo-FS with a dissociation constant of 6-7 microM, which is 2 orders of magnitude higher than the KD value for riboflavin . For the steady-state kinetic analysis, this represents reversible binding of FMN(out) in the phosphorylation cycle (cycle I), which effectively inhibits catalysis in the adenylylation cycle (cycle II). Poult Sci, 1998 Jul, 77(7), 944 - 9 Heterogeneity of staphylococci and other bacteria isolated from six-week-old broiler chickens; Awan MA et al.; In broiler operations, various health problems develop during the final 2 wk of the growing period, resulting in increased mortality and condemnation losses . At this stage, sickly birds were found to be systemically infected by various bacteria regardless of varied clinical signs, and the purpose of this study was to carry out thorough microbiological investigations on this problem . Thirty-one 6-wk-old broilers showing signs of illness were obtained from three farms, and bacterial isolations were carried out from the blood, liver, and hock joint . Bacteria were isolated from 87, 90, and 71% of the blood, liver, and hock joint samples, respectively . Mean bacterial counts in log10 of the blood (per milliliter) and liver (per gram) were 2.15 and 2.93, respectively . Among 132 bacterial isolates, major species were Staphylococcus (60%), Corynebacterium (18%), Escherichia coli (5%), and Stomatococcus (4%) . Among 79 Staphylococcus isolates, 77 were coagulase-negative . Major species of staphylococci were S . lentus (19%), S . simulans (18%), S . cohnii (13%), S . gallinarum (10%), and S . captis (7%) . In addition, six species of gram-positive and five species of gram-negative organisms were isolated . Thus, the apparent systemic infections were not caused by predominant pathogenic bacterial species, and adequately described as mixed infections . There were some significant relationships between isolated bacterial species and sampling sites, suggesting that certain organisms were abundant in the environment of a particular poultry house . These results indicate that systemic infections in market age broilers are caused by mixed bacterial species and suggest that they are caused by suppressed host antibacterial systems rather than pathogenic factors of microorganisms. Eur J Biochem, 1998 May 15, 254(1), 96 - 102 Carbon-flux distribution in the central metabolic pathways of Corynebacterium glutamicum during growth on fructose; Dominguez H et al.; Growth of Corynebacterium glutamicum on fructose was significantly less than that obtained on glucose, despite similar rates of substrate uptake . This was in part due to the production of overflow metabolites (dihydroxyacetone and lactate) but also to the increased production of CO2 during growth on fructose . These differences in carbon-metabolite accumulation are indicative of a different pattern of carbon-flux distribution through the central metabolic pathways . Growth on glucose has been previously shown to involve a high flux (> 50% of total glucose consumption) via the pentose pathway to generate anabolic reducing equivalents . NMR analysis of carbon-isotope distribution patterns of the glutamate pool after growth on 1-13C- or 6-13C-enriched fructose indicates that the contribution of the pentose pathway is significantly diminished during exponential growth on fructose with glycolysis being the predominant pathway (80% of total fructose consumption) . The increased flux through glycolysis during growth on fructose is associated with an increased NADH/NAD+ ratio susceptible to inhibit both glyceraldehyde-3-phosphate dehydrogenase and pyruvate dehydrogenase, and provoking the overflow of metabolites derived from the substrates of these two enzymes . The biomass yield observed experimentally is higher than can be estimated from the apparent quantity of NADPH associated with the pentose pathway and the flux through isocitrate dehydrogenase, suggesting an additional reaction yielding NADPH . This may involve a modified tricarboxylic acid cycle involving malic enzyme, expressed to significantly higher levels during growth on fructose than on glucose, and a pyruvate carboxylating anaplerotic enzyme. J Clin Microbiol, 1998 Jul, 36(7), 2087 - 8 Coryneform bacteria in throat cultures of healthy individuals; von Graevenitz A et al.; Throat swabs from 113 healthy individuals from Hamburg, Germany, and Zurich, Switzerland, were investigated for coryneform bacteria with nonselective and selective media . Ninety specimens contained 123 strains . Surprisingly, 76% of them were strains of Corynebacterium durum (47%) and Rothia dentocariosa (29%) . Only two were strains of Corynebacterium pseudodiphtheriticum, and none were strains of C . striatum, C . amycolatum, or C . diphtheriae. Anal Biochem, 1998 Jun 15, 260(1), 24 - 9 Quantitative restriction fragment length polymorphism: a procedure for quantitation of diphtheria toxin gene CRM197 allele; Pushnova EA et al.; Here we present an assay for quantitation of a particular gene allele in DNA mixtures by means of restriction fragment length polymorphism (RFLP) in combination with polymerase chain reaction (PCR) . We applied the quantitative RFLP principle for estimation of the relative amount of diphtheria toxin gene CRM197 allele in Corynebacterium diphtheriae culture DNA samples . The procedure is based on PCR-mediated generation of an artificial AluI restriction site specifically with the CRM197 DNA template . After AluI digestion of the PCR product and polyacrylamide gel electrophoresis of the restriction fragments, the percentage of CRM197 template in the initial DNA sample was determined by scanning a gel negative . The method was shown to give a linear response when applied to template mixtures containing different amounts of CRM197 reference template . For samples where non-CRM197 DNA was detected by AluI RFLP, we designed a further allele-specific PCR assay to determine whether the non-CRM197 template portion was the wild-type toxin gene allele. Biochem Biophys Res Commun, 1998 Jun 29, 247(3), 741 - 5 Cloning of the histidine biosynthetic genes of Corynebacterium glutamicum: organization and sequencing analysis of the hisA, impA, and hisF gene cluster; Jung SI et al.; The hisA and hisF genes of Corynebacterium glutamicum were cloned by transforming histidine auxotrophic Escherichia coli with the genomic DNA library . They are two of the eight genes that participate in the histidine biosynthetic pathway . Cloned DNA fragments containing the genes can also complement hisH and hisI auxotrophs of Escherichia coli, suggesting that the four genes are clustered in the genome . We determined the nucleotide sequences of the minimal fragment containing the hisA and hisF genes, which are separated by the impA gene . The coding regions of the hisA and hisF genes are 245 and 257 amino acids in length with a predicted size of about 26 and 27 kDa, respectively . These are in good agreement with the sizes of proteins expressed in E . coli . A high similarity was observed in comparison of nucleotide sequences of each protein between C . glutamicum and other species, as well as those between hisA and hisF genes of C . glutamicum. Zentralbl Veterinarmed B, 1998 May, 45(4), 209 - 16 Serological studies on Corynebacterium pseudotuberculosis infections in goats using enzyme-linked immunosorbent assay; Sting R et al.; Corynebacterium pseudotuberculosis was isolated from a goat suffering from caseous lymphadenitis and used for preparation of cell wall antigens and exotoxin for detection of specific antibodies in enzyme-linked immunosorbent assay (ELISA) . In immunoblotting sodium dodecyt sulphate (SDS) extracted cell wall antigens revealed molecular weights ranging from 20 to 120 kDa . The raw exotoxin showed molecular weights of 30 and 55 kDa and an inhibition of the haemolysis of a Staphylococcus aureus strain . For validation of the ELISA 109 goats of known clinical status were examined reaching a sensitivity of 96% and a specificity of 76% for this test . No serological reactions showed in 191 goats originating from 11 flocks which never had suffered from caseous lymphadenitis . Using this ELISA test 24 goats originating from flocks suffering from caseous lymphadenitis were examined serologically before and after vaccination with a bacterin . Before vaccination one of the five goats with clinical signs showed no positive reaction in ELISA . After vaccination all 24 animals showed positive reactions . Of a total of 1868 goats sampled in Baden-Wuerttemberg 41 (2.2%) in 22 (10%) flocks showed positive reactions in ELISA. Infect Immun, 1998 Jul, 66(7), 3279 - 89 Human and murine immune responses to a novel Leishmania major recombinant protein encoded by members of a multicopy gene family; Webb JR et al.; Vaccination of BALB/c mice with Leishmania major promastigote culture filtrate proteins plus Corynebacterium parvum confers resistance to infection with L . major . To define immunogenic components of this protein mixture, we used sera from vaccinated mice to screen an L . major amastigote cDNA expression library . One of the immunoreactive clones thus obtained encoded a novel protein of L . major with a molecular mass of 22.1 kDa . The predicted amino acid sequence of this clone exhibited significant homology to eukaryotic thiol-specific-antioxidant (TSA) proteins . Therefore, we have designated this protein L . major TSA protein . Southern blot hybridization analyses indicate that there are multiple copies of the TSA gene in all species of Leishmania analyzed . Northern blot analyses demonstrated that the TSA gene is constitutively expressed in L . major promastigotes and amastigotes . Recombinant TSA protein containing an amino-terminal six-histidine tag was expressed in Escherichia coli with the pET17b system and was purified to homogeneity by affinity chromatography . Immunization of BALB/c mice with recombinant TSA protein resulted in the development of strong cellular immune responses and conferred protective immune responses against infection with L . major when the protein was combined with interleukin 12 . In addition, recombinant TSA protein elicited in vitro proliferative responses from peripheral blood mononuclear cells of human leishmaniasis patients and significant TSA protein-specific antibody titers were detected in sera of both cutaneous-leishmaniasis and visceral-leishmaniasis patients . Together, these data suggest that the TSA protein may be useful as a component of a subunit vaccine against leishmaniasis. Biotechnology (N Y), 1995 Jun, 13(6), 577 - 82 Transgenic canola and soybean seeds with increased lysine; Falco SC et al.; We have increased the lysine content in the seeds of canola and soybean plants by circumventing the normal feedback regulation of two enzymes of the biosynthetic pathway, aspartokinase (AK) and dihydrodipicolinic acid synthase (DHDPS) . Lysine-feedback-insensitive bacterial DHDPS and AK enzymes encoded by the Corynebacterium dapA gene and a mutant E . coli lysC gene, respectively, were linked to a chloroplast transit peptide and expressed from a seed-specific promoter in transgenic canola and soybean seeds . Expression of Corynebacterium DHDPS resulted in more than a 100-fold increase in the accumulation of free lysine in the seeds of canola; total seed lysine content approximately doubled . Expression of Corynebacterium DHDPS plus lysine-insensitive E . coli AK in soybean transformants similarly caused several hundred-fold increases in free lysine and increased total sed lysine content by as much as 5-fold . Accumulation of alpha-amino adipic acid (AA) in canola and saccharopine in soybean, which are intermediates in lysine catabolism, was also observed. Monaldi Arch Chest Dis, 1998 Feb, 53(1), 14 - 22 Corynebacterium parvum pleurodesis and survival is not significantly influenced by pleural pH and glucose level; Bilaceroglu S et al.; This study was carried out in the pulmonary department of a referral training hospital for thoracic medicine and surgery, with the aim of assessing the effects of pH and glucose level of a pleural effusion (PE) on survival and the response to pleurodesis (PD) with Corynebacterium parvum . A prospective study was carried out in 204 patients with recurrent, symptomatic PEs (73 benign, 131 malignant) . Fifty eight per cent of 204 PEs had low pH (< 7.20; 7.01 +/- 0.14) nd glucose levels (< 60 mg.dL-1; 36 +/- 14 mg.dL-1), whereas the remaining 42% had higher pH (> or = 7.20; 7.36 +/- 0.07 and glucose levels (> or = 60 mg.dL-1; 79 +/- 16 mg.dL-1) . PD was attempted twice with 7 mg of C . parvum injected through chest tube in all patients, who were then followed up for the outcome of PD and for survival from the time of PD until death or the closure of the study (August 1996) . Of 204 cases, 201 were evaluable for survival and outcome of PD . In 91% of the low-and 82% of the high-pH/glucose benign PEs, complete PD was achieved while the corresponding values for the malignant PEs were 79% and 87%, respectively (p > 0.05) . Six per cent of low-and 8% of high-pH benign PEs, and 13% of low- and 9% of high-pH malignant PEs were palliated with partial PD . Failures were 3% and 10% in the low- versus high-pH benign groups, and 8% and 4% in the low- versus high-pH malignancies, respectively . All 201 cases maintained the immediate post-PD outcome throughout the follow-up . Average survival was 21.8 months in high-pH benign PEs versus 21.1 months in low-pH benign PEs, and 9.9 versus 8.7 months, in high- and low-pH malignant PEs, respectively (p > 0.05) . We deduce that, regarding survival and the response to pleurodesis with Corynebacterium parvum, there is no significant difference between low- and high-pH/glucose pleural effusions in malignant, or benign cases. Aust Vet J, 1998 May, 76(5), 335 - 8 Antibiotics for the preservation of koala (Phascolarctos cinereus) semen; Johnston SD et al.; AIM: To determine the normal microbial flora of the koala ejaculate and prepuce in order to select appropriate antibiotics for addition into diluents designed for the preservation of semen . PROCEDURE: Bacteriological samples of the koala prepuce (n = 12) and ejaculate (n = 20) were submitted for microbial culture and sensitivity testing . Microbial flora of ejaculates collected by electroejaculation and artificial vagina were compared . The effects of varying concentrations of penicillin G and gentamicin on sperm motility and on the growth of bacteria in diluted semen stored at room temperature and 16 degrees C over a 24 h period were investigated . RESULTS: A range of bacteria was isolated from the koala prepuce and ejaculate . The predominant organisms in semen collected by electroejaculation and artificial vagina were Corynebacterium spp, none of which could be assigned to any recognised species . The addition of penicillin G and gentamicin to a PBS-based diluent at dose rates of 1000 to 2000 IU/mL and 100 to 200 micrograms/mL respectively, resulted in no adverse effect on sperm motility over a 24 h incubation period . Penicillin G (1000 IU/mL) and gentamicin (100 micrograms/mL) prevented growth of bacterial contaminants in diluted koala semen . CONCLUSION: By controlling the growth of bacteria in extended koala semen, penicillin G and gentamicin are likely to lengthen the period by which spermatozoa can be stored at 16 degrees C and reduce the possibility of disease transmission during artificial insemination procedures. J Urol, 1998 Jul, 160(1), 3 - 9 Encrusted cystitis and pyelitis; Meria P et al.; PURPOSE: Encrusted cystitis and pyelitis are chronic inflammations of the bladder and collecting system associated with mucosal encrustations induced by urea splitting bacteria . We review these infectious diseases . MATERIALS AND METHODS: A literature search was performed of the MEDLINE database from 1985 to 1997 . Additional articles published before 1985 were also selectively included . RESULTS: Most of the articles were case reports or short series . During the last 10 years increasing numbers of cases have been diagnosed, especially in immunodepressed patients, and particularly in renal transplant recipients . Many bacteria have been demonstrated in this infection but Corynebacterium group D2 is currently the most frequent . The development of encrusted cystitis or pyelitis requires the presence of specific bacteria with an alkaline urine, a preexisting urological procedure and a clinical context predisposing to infection . Clinical diagnosis can be difficult but the presence of alkaline urine containing abundant calcified mucopurulent debris is highly suggestive . Demonstration of the bacteria requires prolonged cultures in enriched media . Treatment is based on adapted antibiotic therapy, acidification of urine and excision of plaques of calcified encrustation . The consequences of treatment failure are serious and can result in graft nephrectomy in kidney transplant recipients . CONCLUSIONS: Early clinical and bacterial diagnosis of encrusted cystitis and pyelitis could improve the prognosis of these infectious diseases. Proc Natl Acad Sci U S A, 1998 Jun 9, 95(12), 6768 - 73 Crystal structure of 2,5-diketo-D-gluconic acid reductase A complexed with NADPH at 2.1-A resolution; Khurana S et al.; The three-dimensional structure of Corynebacterium 2, 5-diketo-D-gluconic acid reductase A (2,5-DKGR A; EC 1.1.1.-), in complex with cofactor NADPH, has been solved by using x-ray crystallographic data to 2.1-A resolution . This enzyme catalyzes stereospecific reduction of 2,5-diketo-D-gluconate (2,5-DKG) to 2-keto-L-gulonate . Thus the three-dimensional structure has now been solved for a prokaryotic example of the aldo-keto reductase superfamily . The details of the binding of the NADPH cofactor help to explain why 2,5-DKGR exhibits lower binding affinity for cofactor than the related human aldose reductase does . Furthermore, changes in the local loop structure near the cofactor suggest that 2,5-DKGR will not exhibit the biphasic cofactor binding characteristics observed in aldose reductase . Although the crystal structure does not include substrate, the two ordered water molecules present within the substrate-binding pocket are postulated to provide positional landmarks for the substrate 5-keto and 4-hydroxyl groups . The structural basis for several previously described active-site mutants of 2,5-DKGR A is also proposed . Recent research efforts have described a novel approach to the synthesis of L-ascorbate (vitamin C) by using a genetically engineered microorganism that is capable of synthesizing 2,5-DKG from glucose and subsequently is transformed with the gene for 2,5-DKGR . These modifications create a microorganism capable of direct production of 2-keto-L-gulonate from D-glucose, and the gulonate can subsequently be converted into vitamin C . In economic terms, vitamin C is the single most important specialty chemical manufactured in the world . Understanding the structural determinants of specificity, catalysis, and stability for 2,5-DKGR A is of substantial commercial interest. J Am Vet Med Assoc, 1998 Jun 1, 212(11), 1765 - 8 Evaluation of a commercially available vaccine against Corynebacterium pseudotuberculosis for use in sheep; Piontkowski MD et al.; OBJECTIVE: To evaluate the efficacy of a commercially available bacterin-toxoid vaccine for preventing Corynebacterium pseudotuberculosis-induced abscesses in sheep . DESIGN: Prospective randomized controlled trial . ANIMALS: 31 mixed-breed sheep seronegative for C pseudotuberculosis . PROCEDURE: Sheep were randomly assigned to vaccinate (n = 20) or nonvaccinate (11; control) groups . Sheep in the vaccinate group received 2 doses of serial A or serial B bacterin-toxoid vaccine at 4-week intervals . Serologic testing was conducted after vaccination to document an antibody response to vaccination . All sheep were challenge inoculated with virulent C pseudotuberculosis organisms 32 weeks after the second vaccination . Twenty weeks after challenge inoculation, all sheep were examined for external and internal abscesses secondary to C pseudotuberculosis infection . RESULTS: Vaccinated sheep developed an antibody response to both components of the vaccine, as measured by use of ELISA tests . After challenge inoculation, vaccinated sheep had significantly less external, internal, and total abscesses than control sheep . CLINICAL IMPLICATIONS: Vaccination of sheep with a commercially available bacterin-toxoid against C pseudotuberculosis could substantially decrease the prevalence and number of abscesses that form secondary to C pseudotuberculosis infection. J Bacteriol, 1998 Jun, 180(12), 3233 - 6 The Arcanobacterium (Actinomyces) pyogenes plasmid pAP1 is a member of the pIJ101/pJV1 family of rolling circle replication plasmids; Billington SJ et al.; The 2.4-kb plasmid pAP1 from Arcanobacterium (Actinomyces) pyogenes had sequence similarity within the putative replication protein and double-stranded origin with the pIJ101/pJV1 family of plasmids . pJGS84, a derivative of pAP1 containing a kanamycin resistance gene, was able to replicate in Escherichia coli and Corynebacterium pseudotuberculosis, as well as in A . pyogenes . Detection of single-stranded DNA intermediates of pJGS84 replication suggested that this plasmid replicates by the rolling circle mechanism. J Bacteriol, 1998 Jun, 180(12), 3159 - 65 Different modes of diaminopimelate synthesis and their role in cell wall integrity: a study with Corynebacterium glutamicum; Wehrmann A et al.; In eubacteria, there are three slightly different pathways for the synthesis of m-diaminopimelate (m-DAP), which is one of the key linking units of peptidoglycan . Surprisingly, for unknown reasons, some bacteria use two of these pathways together . An example is Corynebacterium glutamicum, which uses both the succinylase and dehydrogenase pathways for m-DAP synthesis . In this study, we clone dapD and prove by enzyme experiments that this gene encodes the succinylase (M(r) = 24082), initiating the succinylase pathway of m-DAP synthesis . By using gene-directed mutation, dapD, as well as dapE encoding the desuccinylase, was inactivated, thereby forcing C . glutamicum to use only the dehydrogenase pathway of m-DAP synthesis . The mutants are unable to grow on organic nitrogen sources . When supplied with low ammonium concentrations but excess carbon, their morphology is radically altered and they are less resistant to mechanical stress than the wild type . Since the succinylase has a high affinity toward its substrate and uses glutamate as the nitrogen donor, while the dehydrogenase has a low affinity and incorporates ammonium directly, the m-DAP synthesis is another example of twin activities present in bacteria for access to important metabolites such as the well-known twin activities for the synthesis of glutamate or for the uptake of potassium. Am Fam Physician, 1998 May 15, 57(10), 2424 - 32 Skin and wound infections: an overview; O'Dell ML; Skin infections are common and may be caused by bacteria, fungi or viruses . Breaks in the skin integrity, particularly those that inoculate pathogens into the dermis, frequently cause or exacerbate skin infections . Bacterial skin infections caused by corynebacteria include erythrasma, trichomycosis axillaris and pitted keratolysis . Staphylococci may cause impetigo, ecthyma and folliculitis . Streptococcal skin infections include impetigo and erysipelas . Human papillomavirus skin infections present as several different types of warts, depending on the surface infected and its relative moisture, and the patterns of pressure . The many dermatomycoses (skin infections caused by fungi or yeasts) include tinea capitis, tinea barbae, tinea cruris, tinea manus, tinea pedis and tinea unguium (onychomycosis) . Candidal infections occur in moist areas, such as the vulva, mouth, penis, skinfolds and diaper area . Wounds caused by wood splinters or thorns may result in sporotrichosis . Animal bites may result in complex, serious infections, requiring tetanus and, possibly, rabies prophylaxis in addition to appropriate antibiotic therapy. Arch Microbiol, 1998 Apr, 169(4), 303 - 12 Isoleucine uptake in Corynebacterium glutamicum ATCC 13032 is directed by the brnQ gene product; Tauch A et al.; By complementation analysis of an isoleucine-uptake-deficient Escherichia coli strain, it was shown that a 1.6-kb HindIII-StuI fragment of Corynebacterium glutamicum ATCC 13032, located downstream of the aecD gene, encodes an isoleucine uptake system . Sequence analysis revealed that the complementing fragment carried an open reading frame, termed brnQ, that encodes a protein with sequence similarities to branched-chain amino acid carriers of gram-positive and gram-negative bacteria . The brnQ gene specifies a predominantly hydrophobic protein of 426 amino acid residues with a calculated molecular mass of 44.9 kDa . A topology prediction by neural network computer analysis suggests the existence of 12 hydrophobic segments that most probably form transmembrane alpha-helices . A C . glutamicum mutant strain harboring a defined deletion of brnQ in the chromosome showed a considerably lower isoleucine uptake rate of 0.04 nmol min-1 mg (dry mass)-1 as compared to the wild-type strain rate of 1.2 nmol min-1 mg (dry mass)-1 . Overexpression of brnQ by means of a tac promotor resulted in an elevated uptake rate for isoleucine of 11.3 nmol min-1 mg (dry mass)-1 . Evidently, the brnQ gene encodes the only transport system in C . glutamicum directing isoleucine uptake. Pediatr Infect Dis J, 1998 May, 17(5), 391 - 7 The immunogenicity of Haemophilus influenzae type b conjugate vaccines in children born to human immunodeficiency virus-infected women . Women and Infants Transmission Study Group; Read JS et al.; BACKGROUND: Immunocompromise caused by HIV-1 infection increases the importance of receipt of routine childhood vaccines to prevent infections such as invasive Haemophilus influenzae type B (Hib) disease . The objectives of the study were to evaluate the immunogenicity of Hib conjugate vaccines among HIV-infected children according to clinical and immunologic disease progression as well as viral load . METHODS: The concentration of antibody to polyribosylribitol phosphate (PRP) was measured at approximately 9 and 24 months of age in plasma specimens from children of HIV-infected women enrolled in the Women and Infants Transmission Study . RESULTS: Among 227 children (35 HIV-infected, 192 uninfected) at the 9-month study visit who were known to have received age-appropriate immunization with CRM197 mutant Corynebacterium diphtheriae protein-conjugated Hib vaccine, geometric mean antibody concentrations were lower among HIV-infected children (1.64 microg/ml) than among uninfected children (2.70 microg/ml), although the difference was not statistically significant . Anti-PRP antibody concentrations did not vary significantly among these HIV-infected children with predominantly mild-moderate disease progression according to clinical category, immunologic stage or viral load (P > or = 0.48) . The proportion of children with antibody concentrations > or = 1.0 microg/ml did not vary significantly according to HIV infection status (73% uninfected, 74% infected) or, if infected, clinical or immunologic disease progression or viral load . Similar results were obtained among 127 children (17 HIV-infected, 110 uninfected) eligible for analysis at the 24-month study visit . Changes in antibody concentrations over time (between 9 and 24 months of age) did not differ significantly among 10 HIV-infected as compared with 72 uninfected children (P=0.81) . CONCLUSIONS: These results suggest that HIV-infected children with predominantly mild-moderate disease progression respond reasonably well in terms of a quantitative antibody response to Hib conjugate vaccines during the first 2 years of life . Research to further characterize the immune response to Hib conjugate vaccines and to further delineate the "durability" of anti-PRP antibody concentrations beyond 2 years of life should be pursued. Aust Dent J, 1998 Apr, 43(2), 128 - 30 ADRF Trebitsch Scholarship . The microbial contamination of toothbrushes . A pilot study; Taji SS et al.; Ten individuals were each supplied with a new toothbrush of the same type and brand, together with identical tubes of fluoridated toothpaste . After a three-week period, during which subjects were asked to follow their usual oral hygiene practices, the toothbrushes were collected and assayed for microbial contamination using a range of selective growth media . The total microbial load per toothbrush was found to be 10(4) to 10(5) colony forming units . Staphylococci were found on all toothbrushes and streptococci on all but one . These two genera were also quantitatively dominant . Candida, corynebacteria, pseudomonads and coliforms were identified in 70, 60, 50 and 30 per cent of toothbrushes, respectively . However, mutans streptococci, lactobacilli and black-pigmented Gram-negative anaerobic rods were not detected on any of the toothbrushes . For each individual, information on variables such as toothbrush rinsing practices and post-brushing storage methods and environment was collected . No obvious relationship between such variables and microbial load was apparent but it is suggested that more extensive studies are needed, taking into account additional parameters such as age and degree of toothbrush wear and the use of pre-brushing mouthwashes. Arq Gastroenterol, 1997 Jul-Sep, 34(3), 157 - 62 Kupffer cell activation with BCG . Corynebacterium parvum or zymosan protects against acute liver injury induced by carbon tetrachloride in rats; Pereira FE et al.; Kupffer cells have been implicated in the pathogenesis of liver injury, but there is controversy about the effects of activation of these cells on the hepatotoxicity of chemicals and endotoxin . It has been shown that injection of Corynebacterium parvum in rats induces macrophage activation that protects against toxic effects of carbon tetrachloride and acetaminophen, five days after injection, and this protection is due to inhibition of microsomal oxidizing enzymes and increased production of glutathion . To verify if the protective effect occurs soon after Kupffer cell activation, with different activators, male albino rats were treated with intravenous injection of BCG (0.5 ml with 7.5.10(8) bacilli), Corynebacterium parvum (30 mg/kg) or zymosan (7.5.10(6) yeast cells) . Fourty-eight hours after the injection of one of the macrophage activators, the animals and rats treated with intravenous injection of saline (controls) received carbon tetrachloride by subcutaneous route (1 ml/kg of CCl4, 3:1 in soybean oil) . Fourty-eight hours after the animals were killed after ether anesthesia and fragments of the liver were fixed, paraffin embedded and the sections stained with hematoxylin and eosin . A Weibel grid with 168 points was used to estimate the percent volume of necrosis and severe hydropic degeneration . The results showed that the volume density of necrosis and severe hydropic degeneration were significatively lesser in rats treated with the three Kupffer cells activators . The protection was greater with BCG and Corynebacterium parvum than with zymosan . These results confirm that activation of Kupffer cell with three different activators can induce protection against liver cell injury produced by carbon tetrachloride in rats soon as 48 h after injection of activators. Appl Environ Microbiol, 1998 Jun, 64(6), 2295 - 300 Computation of the electrical double layer properties of semipermeable membranes in multicomponent electrolytes Wasserman E, Felmy AR. A methodology is presented for calculating of the surface potential, Donnan potential, and ion concentration profiles for semipermeable microbial membranes that is valid for an arbitrary electrolyte composition . This model for surface potential, Donnan potential, and charge density was applied to recently reported experimental data for gram-positive bacteria, including Bacillus brevis, Rhodococcus opacus, Rhodococcus erythropolis, and Corynebacterium species . These calculations show that previously unconsidered trace amounts of divalent and trivalent cations at very low concentrations (10(-6) M) can have significant effects on the calculated surface and Donnan potentials, at ionic strengths of I </= 0.01 M, and that these effects need to be considered in accurate modeling of microbial surface . In addition, the calculated ion concentration profiles show that owing to the relatively high surface charges that can develop in microbial membranes, electrostatic effects can act to significantly concentrate divalent (factors of 5 x 10(3)) and trivalent (factors of 2 x 10(4)) cations within the bacterial cell wall . Comparison of the calculated concentration factors with those derived from experiments shows that a significant fraction of the uptake of metal by bacteria can be explained by the proposed electrostatic model. Arch Microbiol, 1998 May, 169(5), 411 - 6 Urea uptake and urease activity in Corynebacterium glutamicum; Siewe RM et al.; When Corynebacterium glutamicum is grown with a sufficient nitrogen supply, urea crosses the cytoplasmic membrane by passive diffusion . A permeability coefficient for urea diffusion of 9 x 10(-7) cm s-1 was determined . Under conditions of nitrogen starvation, an energy-dependent urea uptake system was synthesized . Carrier-mediated urea transport was catalyzed by a secondary transport system linked with proton motive force . With a Km for urea of 9 microM, the affinity of this uptake system was much higher than the affinity of urease towards its substrate (Km approximately 55 mM urea) . The maximum uptake velocity depended on the expression level and was relatively low {2-3.5 nmol min-1 (mg dry wt.)-1}. J Neuroimmunol, 1998 Apr 1, 84(1), 92 - 104 Adhesion molecule phenotype of T lymphocytes in inflamed CNS; Engelhardt B et al.; The phenotype of T cells in the central nervous system (CNS) in two models of chronic inflammation (experimental allergic encephalomyelitis and Corynebacterium parvum-induced inflammation) was compared to that of T cells in gut and chronically inflamed subcutaneous tissue and lung . CNS T cells display a similar phenotype in both inflammatory models, and are phenotypically unique compared to T cells from the other inflamed tissues . T cells from inflamed CNS are mainly CD4+ and are the only population examined that express a typical activated/memory phenotype: CD44high/LFA-1high/ICAM-1high/CD45RBlow . The CNS T cells are alpha4beta7-integrin(negative), but express alpha4-integrin and activated beta1 integrin, suggesting expression of the alpha4beta1-heterodimer in an activated state . In contrast, most T cells in gut express low levels of activated beta1 integrin . The CNS T cells lack expression of alpha6 and alphaE integrin chains and L-selectin . In inflamed CNS and inflamed subcutaneous tissue, approximately 50% of T cells express high affinity ligands for P-selectin while fewer than 10% express high affinity ligands for E-selectin . In summary, our data show that, independent of the inflammatory stimulus, T cells recruited into the inflamed CNS are phenotypically distinct from T cells in other inflamed tissues . This finding leads us to hypothesize the existence of a phenotypically distinct 'CNS-seeking' T lymphocyte population. Rev Assoc Med Bras, 1997 Oct-Dec, 43(4), 326 - 34 {Bacteremia after endoscopic retrograde cholangiopancreatography with and without therapeutic procedure: frequency, associated factors and clinical significance}; Campos GM et al.; PURPOSE: To determine the frequency, associate factors and clinical features of bacteremia in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP), with or without therapeutic procedures . METHODS: Prospectively, 42 consecutives patients undergoing 46 endoscopic retrograde cholangiopancreatographies (ERCPs) from August to December 1994 were analyzed . The search for bacteremia was done by drawing 6 blood samples for cultures from peripheral blood . Two blood samples were collected before the ERCP and 4 of them after . The bottles used for cultures were Bactec bottles . The bottles were incubated in the Bactec 9240 system, and eventual bacteria detect were identificated by the manual routine of the laboratory and also with the autoScan/Microscan system . RESULTS: All blood cultures obtained before the ERCPs were negatives . Bacteremia were detected after 7 endoscopic procedures . In two episodes of bacteremia, the microorganism identified (Staphylococcus epidermidis) was considered to be a contaminant . The other 5 episodes of bacteremia were considered true bacteremia (frequency- 10.9%), and the microorganisms identified were: Streptococcus viridans, Corynebacterium sp., Enterobacter cloacae, Klebsiello oxytoca and Enterobacter aerogenes . This episodes were more frequent in the blood cultures obtained immediately after the ERCPs (p < 0.05), and occurred exclusively in the patients who were not receiving antibiotics (p = 0.0192) . Clinical manifestation of the episodes of bacteremia were not detected . CONCLUSION: The episodes of bacteremia occurred exclusively in the patients who were not receiving antibiotics, were transient and completely no symptomatic. Antimicrob Agents Chemother, 1998 May, 42(5), 1127 - 32 Activities of HMR 3004 (RU 64004) and HMR 3647 (RU 66647) compared to those of erythromycin, azithromycin, clarithromycin, roxithromycin, and eight other antimicrobial agents against unusual aerobic and anaerobic human and animal bite pathogens isolated from skin and soft tissue infections in humans; Goldstein EJ et al.; The activities of HMR 3004 and HMR 3647 and comparator agents, especially macrolides, were determined by the agar dilution method against 262 aerobic and 120 anaerobic strains isolated from skin and soft tissue infections associated with human and animal bite wounds . HMR 3004 and HMR 3647 were active against almost all aerobic and fastidious facultative isolates (MIC at which 90% of the isolates are inhibited {MIC90}, < or = 0.5 and 1 microg/ml, respectively) and against all anaerobes {Bacteroides tectum, Porphyromonas macacae (salivosa), Prevotella heparinolytica, Porphyromonas sp., Prevotella sp., and peptostreptococci} at < or = 0.25 and < or = 0.5 microg/ml, respectively, except Fusobacterium nucleatum (HMR 3004, MIC90 = 16 microg/ml; HMR 3647, MIC90 = 8 microg/ml) and other Fusobacterium species (MIC90, 1 and 2 microg/ml, respectively) . In general, HMR 3004 and HMR 3647 were more active than any of the macrolides tested . Azithromycin was more active than clarithromycin against all Pasteurella species, including Pasteurella multocida subsp . multocida, Eikenella corrodens, and Fusobacterium species, while clarithromycin was more active than azithromycin against Corynebacterium species, Weeksella zoohelcum, B . tectum, and P . heparinolytica. Antimicrob Agents Chemother, 1998 May, 42(5), 1028 - 33 In vitro susceptibilities of aerobic and facultative non-spore-forming gram-positive bacilli to HMR 3647 (RU 66647) and 14 other antimicrobials; Soriano F et al.; The comparative in vitro activity of the ketolide HMR 3647 (RU 66647) and those of structurally related macrolide-lincosamide-streptogramin compounds (erythromycin, roxithromycin, azithromycin, clarithromycin, josamycin, lincomycin, pristinamycin, and quinupristin-dalfopristin) as well as those of benzylpenicillin, doxycycline, vancomycin, teicoplanin, levofloxacin, and rifapentine against 247 aerobic and facultative non-spore-forming gram-positive bacilli were determined by an agar dilution method . The ketolide was active against most organisms tested except Corynebacterium striatum, coryneform CDC group 12, and Oerskovia spp . The frequency of resistance to erythromycin and other macrolides as well as that to lincomycin was high . Pristinamycin and, to a lesser extent, quinupristin-dalfopristin were very active, but resistance to these agents was present in some strains of Rhodococcus equi, Listeria spp., C . striatum, Erysipelothrix rhusiopathiae, and Oerskovia spp . HMR 3647 was very active against all erythromycin-sensitive and many erythromycin-nonsusceptible strains, especially Corynebacterium minutissimum, Corynebacterium pseudodiphtheriticum, Corynebacterium amycolatum, and Corynebacterium jeikeium . In vitro resistance to benzylpenicillin was common, but doxycycline, vancomycin, and teicoplanin were very active against most organisms tested except E . rhusiopathiae, against which glycopeptide antibiotics were not active . The in vitro activity of levofloxacin was remarkable, but resistance to this agent was common for C . amycolatum, Corynebacterium urealyticum, C . jeikeium, and Oerskovia spp . strains . Rifapentine was also very active in vitro against many organisms, but resistance to this agent was always present in E . rhusiopathiae and was very common in C . striatum and C . urealyticum. Acta Vet Scand, 1998, 39(1), 109 - 17 Control of caprine arthritis-encephalitis virus and Corynebacterium pseudotuberculosis infection in a Norwegian goat herd; Nord K et al.; A control programme for caprine arthritis-encephalitis virus (CAEV) and Corynebacterium pseudotuberculosis (C . pseudotuberculosis) infection was established in a Norwegian goat herd comprising approximately 100 milking goats . The herd seroprevalences of antibodies against CAEV and C . pseudotuberculosis were 97% and 94%, respectively . Kids were removed from the infected flock at birth, avoiding any contact between dam and kid . The kids were kept completely segregated from the seropositive flock and fed cow's colostrum and milk . A seronegative flock was established, based on the removed kids and their offspring . Goasts belonging to the seronegative flock were allowed to kid naturally and to mother their kids . The seropositive flock was slaughtered during the second year of the control programme . After washing and disinfection, housing systems and nearby outdoor premises were left empty for 3 months . Of 230 goats examined for antibodies against CAEV with ELISA regularly during 3 years of the control program, altogether 6 were found to be seropositive, while for 10 the result was indeterminate . All 16 animals were immediately culled . During the third year of the control programme, all goats were examined and proved negative for antibodies against C . pseudotuberculosis by a haemolysis inhibition test . Clinical examination revealed no signs of CAE or caseous lymphadenitis. Semin Surg Oncol, 1998 Jun, 14(4), 302 - 10 Adjuvant therapy of melanoma; Agarwala SS et al.; Patients with AJCC Stage IIB and III melanoma have a poor 5-year survival rate which has been the driving force behind attempts to find an effective adjuvant therapy for this stage of disease that would effectively reduce relapse and improve survival . Immunotherapy with bacillus Calmette-Guerin (BCG), Corynebacterium parvum, and levamisole have not been successful in achieving this goal, nor have trials with chemotherapy in the adjuvant setting, including high-dose chemotherapy with autologous bone marrow transplantation . The recent Eastern Cooperative Oncology Group (ECOG) 1684 study showed significant improvement in relapse-free and overall survival with high doses of alpha interferon (IFNalpha) given for 1 year . Lower dosages of IFNalpha have to date been unsuccessful in impacting upon long-term survival . Recent data with vaccines have been encouraging, and the GM2-KLH vaccine is the focus of ongoing intergroup study comparing this treatment with IFNalpha in resected Stage IIB and III melanoma . The various regimens are reviewed in this article. Genetika, 1998 Mar, 34(3), 438 - 41 {Construction and characteristics of a cosmid library of genes of the bacterium Cornyebacterium glutamicum ATSS13032}; Bukanov NO et al.; A representative genomic library of the Corynebacterium glutamicum ATCC 13032 genes in a cosmid vector Lorist6 was created . The cosmids contain inserts of bacterial DNA obtained by partial digestion with the Sau3A I restrictase . Five hundred and thirty individual primary recombinant clones were transferred into the wells of microtiter plates, where they are now being preserved . The average size of the bacterial DNA inserts determined via a sum of restriction fragment sizes of recombinant molecules is about 38 kb . The capacity of the obtained gene library is 8.4 equivalents of the C . glutamicum genome, i.e., every fragment of the genome is on average represented by eight clones and is presented in at least one clone with the probability > 99% . Clone grids (sets of recombinant clones located on the hybridization membrane in regular and reproducible order) were created . Specificity of the created clone library and its representativeness were confirmed experimentally by hybridization of clone grids with DNA probes corresponding to unique regions of the Corynebacterium genome . A plasmid containing the pheA prephenate dehydratase gene, olygonucleotide corresponding to the lysC gene, and the 21 RNA probe obtained from the insert ends in different cosmids were used as probes . The created set of clones allows the construction of a cosmid contig overlapping the C . glutamicum genome and a physical genetic map on its base. Genitourin Med, 1997 Dec, 73(6), 477 - 80 Nasopharyngeal flora in HIV seropositive men who have sex with men; Carlin EM et al.; OBJECTIVES: To assess, in men who were infected with the human immunodeficiency virus (HIV) and who identified themselves as having had sex with men; the nasopharyngeal prevalence of Neisseria gonorrhoeae, N meningitidis, Corynebacterium diphtheriae, and candida species; oral sexual behaviour; the relation between oral flora and oral sexual behavior . METHOD: Nasopharyngeal swabs were taken from HIV seropositive men for culture . The men were also asked to complete a self administered questionnaire . RESULTS: 390 men were recruited; 286 (73.3%) provided nasopharyngeal samples and questionnaires; 41 (10.5%) provided nasopharyngeal samples only; 63 (16.2%) provided questionnaires only . From the 327 nasopharyngeal samples N meningitidis was cultured in 49 (15%) and candida species in 165 (50.5%) . Cultures for N gonorrhoeae and C diphtheriae were all negative . Data from the 349 completed questionnaires indicated that 285 men were practising oro-penile sex, over 90% did not consistently use condoms; 150 men were practising oro-anal sex, one used dental dams . In those providing both nasopharyngeal samples and sexual behaviour data meningococcal carriage was identified in 40 (17.5%) of the 228 men practising receptive oro-penile sex, compared with one (2.3%) of the 43 non-practisers (p < 0.025); in 21 (20%) of the 105 men practising insertive oro-anal sex, compared with 17 (12.5%) of the 136 non-practisers (p = 0.12) . No correlation was identified between yeast carriage and oro-genital sex . Conclusion: Oro-genital sex, usually without barrier protection, is common among HIV infected men who have sex with men . It appears to be associated with increased meningococcal carriage but is autonomous to candida species isolation . Routine screening for nasopharyngeal N gonorrhoeae is not deemed necessary. Microbiology, 1998 Apr, 144 ( Pt 4), 915 - 27 Pyruvate carboxylase from Corynebacterium glutamicum: characterization, expression and inactivation of the pyc gene; Peters-Wendisch PG et al.; In addition to phosphoenolpyruvate carboxylase (PEPCx), pyruvate carboxylase (PCx) has recently been found as an anaplerotic enzyme in the amino-acid-producing bacterium Corynebacterium glutamicum . Using oligonucleotides designed according to conserved regions of PCx amino acid sequences from other organisms, a 200 bp fragment central to the C . glutamicum PCx gene (pyc) was amplified from genomic DNA by PCR . This fragment was then used to identify and to subclone the entire C . glutamicum pyc gene . The cloned pyc gene was expressed in C . glutamicum, as cells harbouring the gene on plasmid showed four- to fivefold higher specific PCx activities when compared to the wild-type (WT) . Moreover, increased PCx protein levels in the pyc-plasmid-carrying strain were readily detected after SDS-PAGE of cell-free extracts . DNA sequence analysis of the pyc gene, including its 5' and 3' flanking regions, and N-terminal sequencing of the pyc gene product predicts a PCx polypeptide of 1140 amino acids with an M(r) of 123070 . The amino acid sequence of this polypeptide shows between 62% and 45% identity when compared to PCx enzymes from other organisms . Transcriptional analyses revealed that the pyc gene from C . glutamicum is monocistronic (3.5 kb mRNA) and that its transcription is initiated at an A residue 55 bp upstream of the translational start . Inactivation of the chromosomal pyc gene in C . glutamicum WT led to the absence of PCx activity and to negligible growth on lactate, indicating that PCx is essential for growth on this carbon source . Inactivation of both the PCx gene and the PEPCx gene in C . glutamicum led additionally to the inability to grow on glucose, indicating that no further anaplerotic enzymes for growth on carbohydrates exist in this organism. J Wildl Dis, 1998 Apr, 34(2), 397 - 9 Corynebacterial pneumonia in an African hedgehog; Raymond JT et al.; A 3-mo-old, male African hedgehog (Atelerix albiventris) was anorectic and lethargic for a period of 3 days prior to death . Necropys revealed lungs that were diffusely firm, dark red, and dorsally adhered by fibrinous tags to the pericardial sac . Histopathology revealed necrosuppurative bronchopneumonia with pulmonary abscesses and suppurative pericarditis and myocarditis . A Corynebacterium sp . was isolated from the lungs . We believe this is the first reported case of corynebacterial pneumonia in an African hedgehog. J Clin Microbiol, 1998 May, 36(5), 1430 - 2 Identification of Corynebacterium amycolatum and other nonlipophilic fermentative corynebacteria of human origin; Wauters G et al.; Four identification tests, proposed in addition to conventional methods, were evaluated with 320 fermentative nonlipophilic Corynebacterium strains: growth at 20 degrees C, glucose fermentation at 42 degrees C, alkalinization of sodium formate, and acid production from ethylene glycol . These tests were highly discriminant . Corynebacterium amycolatum displayed a unique profile, allowing it to be distinguished from similar species, such as C . xerosis, C . striatum, and C . minutissimum. Diagn Microbiol Infect Dis, 1998 Mar, 30(3), 167 - 72 Use of random amplified polymorphic DNA for rapid molecular subtyping of Corynebacterium diphtheriae; Nakao H et al.; A total of 210 Corynebacterium diphtheriae strains isolated worldwide were assayed by random amplified polymorphic DNA (RAPD) assay . RAPD was as discriminating as standard ribotyping, and in some cases, even further differentiation was obtained . RAPD can rapidly aid clinical and molecular epidemic studies in a simple and cost-effective manner. Int J Syst Bacteriol, 1998 Jan, 48 Pt 1, 69 - 74 Description of some coryneform bacteria isolated from human clinical specimens as Corynebacterium falsenii sp . nov; Sjoden B et al.; Over a five-year period, four strains of a yellowish-pigmented coryneform bacterium were received for identification by the Culture Collection of the University of Goteborg . All strains had been isolated from normally sterile human body fluids . Initial biochemical characterization revealed that all four isolates were very similar, with weak pyrazinamidase and urease activities, as well as slow fermentative acid production from glucose as the most significant phenotypic features which differentiated the strains from all other presently defined corynebacteria . Chemotaxonomic investigations demonstrated that the strains belonged to the genus Corynebacterium . SDS-PAGE of whole-cell proteins suggested that all four strains were representatives of the same species . Comparative 16S rRNA gene sequence analysis unambiguously demonstrated that the four strains were genealogically related and represent a new subline within the genus Corynebacterium for which the designation Corynebacterium falsenii sp . nov . is proposed . The type strain of Corynebacterium falsenii is CCUG 33651. Med Dosw Mikrobiol, 1997, 49(3-4), 131 - 40 {Utilization of Staphylococcus siderophores produced by Corynebacterium and Coryneform bacteria}; Szarapinska-Kwaszewska J et al.; The ability of iron utilizing by means of siderophores produced by donor strains-Coryne-bacterium and coryneform organisms (8 strains) by 24 staphylococcal strains was investigated . All the donor strains synthesized catecholate class siderophores and two strains also hydroxamate class . The majority of staphylococcal strain could utilize these siderophores . Most of strains utilized siderophores from Corynebacterium pseudodiphtheriticum and Corynebacterium aquaticum as well as chelators from plant pathogens-coryneform organisms . Only two staphylococcal strains were not be able to utilize siderophores from all donor strains. Calcif Tissue Int, 1998 Apr, 62(4), 350 - 8 Purification, amino acid sequence, and cDNA sequence of a novel calcium-precipitating proteolipid involved in calcification of corynebacterium matruchotii; van Dijk S et al.; Corynebacterium matruchotii is a microbial inhabitant of the oral cavity associated with dental calculus formation . It produces membrane-associated proteolipid capable of inducing hydroxyapatite formation in vitro . This proteolipid was purified from chloroform:methanol extracts by chromatography on Sephadex LH-20 and migrated on SDS-polyacrylamide gel electrophoresis at 6-9 kDa . Removal of covalently attached acyl moieties by methanolic KOH decreased its molecular mass to approximately 5.5 kDa . The amino acid sequence of the apoproteolipid indicated a peptide of 50 amino acids, a calculated molecular weight of 5354 Da, and an isoelectric point of 4.28 . Sequence analysis revealed an 8 amino acid sequence with homology to human phosphoprotein phosphatase 2A as well as several potential acylation sites and one phosphorylation site . The purified proteolipid induced calcium precipitation in vitro . Deacylation of the proteolipid by hydroxylamine treatment resulted in >50% loss of calcium-precipitating activity, suggesting that covalently attached lipids are required . Degenerate oligonucleotide primers, based on the amino acid sequence, were used to amplify the gene for the 5.5 kDa proteolipid from total chromosomal DNA of C . matruchotii by PCR . A 166 bp cDNA was isolated and sequenced, confirming the amino acid sequence of the proteolipid . Thus, we have sequenced a unique bacterial proteolipid that is involved in the formation of dental calculus by precipitating Ca2+ and possibly in transport of inorganic phosphate, necessary for hydroxyapatite formation. Eur J Biochem, 1998 Mar 15, 252(3), 360 - 71 Effect of reversible reactions on isotope label redistribution--analysis of the pentose phosphate pathway; Follstad BD et al.; The pentose phosphate pathway plays several key roles in metabolism including supply of biosynthetic carbon skeletons and reducing power . Previous research has focused on determining the fluxes through the reactions of this pathway using carbon-labeled substrates and models that make certain assumptions about the reversibility of the transketolase and transaldolase reactions in the nonoxidative pathway . These assumptions, however, have resulted in inconsistencies between the predicted carbon label distributions using these models and those determined experimentally . A general metabolic reaction network model developed in this paper and applied to the pentose phosphate pathway not only incorporates reaction reversibility but also accounts for the effect of individually varying extents of reaction reversibility on labeled carbon fractional enrichment values for intermediate metabolites . In addition, an algorithm is presented that can be used to calculate the three individual transaldolase and transketolase extents of reversibility . The results of this method show that varying extents of reaction reversibility have an observable effect on the metabolite carbon label distributions which can in turn affect flux calculation for other parts of the metabolic network such as the tricarboxylic acid cycle . In addition, the observability of reversibility extent and accuracy of flux calculations depend on the particular choice of metabolite carbon enrichments measured . In particular, {6-13C}hexose 6-phosphate and {4-13C}erythrose 4-phosphate carbon enrichment values resulting from {1-13C}glucose feeding contained more information as compared to those from ribose 5-phosphate . This analysis was applied to literature data of metabolite carbon labeling that resulted from supplying either 13C- or 14C-enriched substrates to several cell types growing under various conditions . The specific activities of metabolite carbon atoms taken from rat epididymal adipose tissue, goosefish islet cells, Corynebacterium glutamicum, and Escherichia coli supplied with either {2-14C}glucose or {1-13C}glucose demonstrate how reversibility is present in the pentose phosphate pathway and the extents of reversibility can be estimated from labeled carbon data sets. Gut, 1998 Feb, 42(2), 266 - 71 Omeprazole induces altered bile acid metabolism; Shindo K et al.; BACKGROUND: It has been reported that the acidity of gastric contents could be an important factor in regulating jejunal flora . AIMS: To investigate the effects of omeprazole induced changes in gastric pH on jejunal flora and bile acid metabolism . METHODS: Twenty one patients with gastric ulcer and 19 healthy volunteers were studied . Deconjugation of bile acids was detected using a bile acid breath test . Jejunal fluid was aspirated using a double lumen tube with a rubber cover on the tip and deconjugation was examined using thin layer chromatography . Fat malabsorption was detected by a triolein breath test . RESULTS: In the bile acid breath test, expired breath samples from all patients and healthy volunteers showed significantly greater 14CO2 specific activity after omeprazole treatment (20 mg/day) than before treatment . Bacterial overgrowth was found in the jejunal fluid and gastric juice of both ulcer patients and healthy volunteers after omeprazole treatment . The following species were identified: Escherichia coli, Candida albicans, enterococcus, Lactobacillus bifidus, Bacteroides vulgatus, B uniformis, Eubacterium lentum, Eu parvum, and Corynebacterium granulosum . All of these species, except E coli and C albicans, deconjugate bile acids . There was a significant correlation between 14CO2 activity and gastric pH, both before and after omeprazole treatment in both groups . The triolein breath test revealed impaired fat absorption in both groups after omeprazole treatment . CONCLUSIONS: Both patients with gastric ulcer and healthy volunteers exhibited increased deconjugation of bile acids caused by bacterial overgrowth in the jejunum and fat malabsorption after omeprazole treatment . The bacterial over-growth consisted of both anaerobes and aerobes with deconjugation ability and was probably associated with an omeprazole induced shift to neutral pH in the gastric juice. Zh Mikrobiol Epidemiol Immunobiol, 1998 Jan-Feb, (1), 17 - 21 {The role of persistence factors in the forming of a microbial biocenosis in the nasal mucosa in staphylococcal bacteria carriers}; Parshuta LI et al.; The microbial biocenosis of the nasal mucosa under normal conditions and in cases of Staphylococcus aureus carriership was studied, taking into account the biological properties of symbionts in 159 persons . In S . aureus carriers the dysbiotic state of the intranasal microflora was established: the decrease of the index of total microbial contamination, the index of contamination with obligate coccal microflora and the index of specific diversity . The factors of the persistence and antagonism of dysbiotic microflora contribute to the mechanism of the development of dysbiosis . In the biocenosis of the carriers obligate microflora strains (Corynebacterium, Micrococcus and coagulase-negative staphylococci) with more pronounced interspecific antagonism or bacteriocinogeny were dominant. Zh Mikrobiol Epidemiol Immunobiol, 1998 Jan-Feb, (1), 13 - 7 {The development of bacterial growth stimulants from plants}; Adlova GP et al.; Dried bacterial growth stimulators in the form of water extracts of wild marjoram were developed; they produced a stimulating effect on Escherichia coli and Streptococcus pyogenes Dick 1 and had no influence on the growth of Corynebacterium xerosis 1911 . The study aimed at finding out the active principle of the stimulator prepared from marjoram extract revealed that under experimental conditions marjoram extract could be divided into 3 fractions of these: fraction 1 contained terpenic hydrocarbons, fraction 2 contained vitamins and vitamin-like substances, fraction 3 contained phenols, flavonoids, phenolcarbonic and fatty acids . Fraction 3 at a concentration of 0.0001% produced the best effect . The stimulating effect of extracts obtained from lime and aspen leaves, haw berries was demonstrated. Zentralbl Veterinarmed B, 1998 Feb, 45(1), 31 - 5 Corynebacterium pseudotuberculosis infection in goats in the Czech Republic; Skalka B et al.; Prevalence of caseous lymphadenitis was studied in goats from six herds in the Czech Republic . Corynebacterium pseudotuberculosis exhibiting typical properties was isolated from clinically affected animals only . Antibodies to C . pseudotuberculosis were identified by means of agar immunodiffusion and the neutralization test using a toxin--phospholipase D (PLD)--as antigen . No clinical manifestations of caseous lymphadenitis or antibodies to C . pseudotuberculosis were found in four herds . In one herd without any history of clinical caseous lymphadenitis, serological positivity was reported in two out of 148 examinations . In a herd with clinical manifestations of caseous lymphadenitis, antibodies were ascertained to a various degree in subsequent samplings (100%, 87% and 64%, respectively) . The importance of serologic examination of caseous lymphadenitis in goat herds is discussed. J Zoo Wildl Med, 1997 Dec, 28(4), 471 - 5 Comparative rectal bacterial flora of four species of flying fox (Pteropus sp.); Heard DJ et al.; The rectal anaerobic and aerobic bacterial flora of four species of flying foxes were determined and compared . Four bacterial species were found in > or = 1 individual from each bat species at a significant (> or = 10%) level of the bacterial population: alpha-hemolytic Streptococcus sp . (41 of 56 bats), Enterococcus sp . (25/56), Escherichia coli (21/ 56), and group D Streptococcus sp., not Enterococcus sp . (9/56) . Five other microbial species were also found in all four flying fox species, but at less significant percentages (found in at least one bat species, > or = 5% and < or = 10% of the recovered microbial population) . These were nonhemolytic Streptococcus sp . (30/56), yeast (26/56), Corynebacterium sp . (25/56), Staphylococcus sp . (25/56), and Staphylococcus aureus (22/56) . The majority of the species found were gram-positive, and only two obligate anaerobes, a Lactobacillus and a Bacteroides sp., were recovered from one bat. Biochemistry, 1998 Mar 10, 37(10), 3278 - 85 Substrate and inhibitor binding sites in Corynebacterium glutamicum diaminopimelate dehydrogenase; Scapin G et al.; The three-dimensional structures of Corynebacterium glutamicum diaminopimelate dehydrogenase as a binary complex with the substrate meso-diaminopimelate (meso-DAP) and a ternary complex with NADP+ and an isoxazoline inhibitor {Abbot, S.D., Lane-Bell, P., Kanwar, P.S.S., and Vederas, J . C . (1994) J . Am . Chem . Soc . 116, 6513-6520} have been solved and refined against X-ray diffraction data to 2.2 A . Diaminopimelate dehydrogenase is a homodimer of approximately 35,000 molecular weight subunits and is the only dehydrogenase present in the bacterial diaminopimelate/lysine biosynthetic pathway . Inhibitors of the enzymes of L-lysine biosynthesis have been proposed as potential antibiotics or herbicides, since mammals lack this metabolic pathway . Diaminopimelate dehydrogenase catalyzes the unique, reversible, pyridine dinucleotide-dependent oxidative deamination of the D-amino acid stereocenter of meso-diaminopimelate to generate L-2-amino-6-oxopimelate . The enzyme is absolutely specific for the meso stereoisomer of DAP and must distinguish between two opposite chiral amino acid centers on the same symmetric substrate . The determination of the three-dimensional structure of the enzyme--meso-diaminopimelate complex allows a description of the molecular basis of this stereospecific discrimination . The substrate is bound in an elongated cavity, in which the distribution of residues that act as hydrogen bond donors or acceptors defines a single orientation in which the substrate may bind in order to position the D-amino acid center of meso-DAP near the oxidized nucleotide . The previously described isoxazoline inhibitor binds at the same site as DAP but has its L-amino acid center positioned where the D-amino acid center of meso-DAP would normally be located, thereby generating a nonproductive inhibitor complex . The relative positions of the N-terminal dinucleotide and C-terminal substrate-binding domains in the diaminopimelate dehydrogenase--NADP+, diaminopimelate dehydrogenase--DAP, and diaminopimelate dehydrogenase--NADP(+)--inhibitor complexes confirm our previous observations that the enzyme undergoes significant conformational changes upon binding of both dinucleotide and substrate. Cornea, 1998 Mar, 17(2), 230 - 2 Mycobacterium chelonei masquerading as Corynebacterium in a case of infectious keratitis: a diagnostic dilemma; Garg P et al.; PURPOSE: The diagnosis of Mycobacterium keratitis can often be missed both clinically and microbiologically and this report highlights one such case . METHODS: Review of medical and microbiological records . RESULTS: We report a case of Mycobacterium keratitis in a 25-year-old man that was misdiagnosed as Corynebacterium keratitis at initial presentation . Presence of partially stained and beaded bacilli in a Gram-stained smear of repeat corneal scrapings raised the suspicion of an unusual organism . Ziehl-Neelsen staining of the decolorized Gram-stained smear and subculture on Lowenstein-Jensen medium