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| 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 helped us to establish the diagnosis . CONCLUSIONS: A high degree of suspicion needs to be maintained, especially in cases in which (a) there is a history of corneal trauma involving a foreign body, (b) the Gram-stained smear of corneal scrapings shows a paucity of organisms and the presence of partially stained and beaded bacilli in the presence of confluent growth of colonies resembling those of Corynebacterium, and (c) a typical corneal feature like "cracked windshield" stromal lesion is seen, to avoid such a misdiagnosis . Inclusion of a Lowenstein-Jensen culture at the initial presentation, especially when the clinical presentation is atypical, as seen in this case, will lead to an early diagnosis. Biochemistry, 1998 Feb 24, 37(8), 2089 - 95 Identification of the covalent flavin attachment site in sarcosine oxidase; Chlumsky LJ et al.; Sarcosine oxidase from Corynebacterium sp . P-1 is a heterotetrameric enzyme (alphabetagammadelta) that contains two noncovalently bound coenzymes (FAD, NAD+) and covalently bound FMN {8alpha-(N3-histidyl)FMN} which is attached to the beta subunit . Chlumsky et al . {(1995) J . Biol . Chem . 270, 18252-18259} tentatively identified His175 as the covalent FMN attachment site in the beta subunit, based on an alignment of the sequence of C . sp . P-1 beta subunit with a highly homologous flavin-containing peptide from another corynebacterial sarcosine oxidase (C . sp . U-96) . To test this hypothesis, His175 in the C . sp . P-1 beta subunit was mutated to an alanine . Unexpectedly, the mutant enzyme was found to contain 1 mol of covalently bound flavin and to exhibit catalytic activity similar to wild-type enzyme . Covalent flavin-containing peptides were isolated from wild-type and mutant enzymes and analyzed by electrospray mass spectrometry . The mass observed for the mutant peptide (1152.4 Da) matched that predicted for an FMN-containing hexapeptide, corresponding to residues 173-178 (1152.1 Da) . In the mutant, this region (HDAVAW) contains a single histidine (His173) which must be the covalent flavin attachment site . The mass observed for the wild-type peptide (1218.6 Da) matched that predicted for an FMN-containing hexapeptide, also corresponding to residues 173-178 in the beta subunit (1218.2 Da) . This region in the wild-type enzyme includes two histidine residues (HDHVAW) . Attempts to sequence the wild-type or mutant peptides by automated Edman degradation were unsuccessful . Instead, the peptide sequences were investigated by collisional-activated dissociation (CAD) and tandem mass spectrometry . The CAD mass spectral data with the mutant peptide confirmed the sequence deduced based on the mass of the intact peptide . The CAD mass spectral results with the wild-type peptide showed that FMN was covalently attached to the N-terminal histidine in the hexapeptide, which corresponds to His173 in the beta subunit. J Antimicrob Chemother, 1997 May, 39 Suppl A, 67 - 8 Minimal inhibitory concentrations and minimal bactericidal concentrations of quinupristin/dalfopristin against clinical isolates of Corynebacterium jeikeium and Listeria monocytogenes; Moore LS et al.; Thirty clinical isolates of Listeria monocytogenes and 30 clinical isolates of Corynebacterium jeikeium were tested against quinupristin/dalfopristin by the NCCLS broth macrodilution technique for determination of MICs . MBC testing was also performed on the 60 isolates . Quinupristin/dalfopristin MIC90 values against L . monocytogenes and C . jeikeium were 1.6 mg/L and 0.2 mg/L, respectively . Quinupristin/dalfopristin was bacteriostatic against these organisms. Vox Sang, 1998, 74(2), 88 - 94 Bacterial contamination of autologous bone marrow: reinfusion of culture-positive grafts does not result in clinical sequelae during the posttransplantation course; Schwella N et al.; OBJECTIVES: Microbiological cultures and posttransplantation course were analyzed in order to investigate the incidence and clinical significance of bacterial contamination of autologous bone marrow (BM) grafts . METHODS: Cultures were obtained from BM after collection, BM concentrate after processing, contaminated/cryopreserved BM at thawing, and from peripheral blood (PB) following autologous BM transplantation (ABMT) . The posttransplantation course of patients grafted with culture-positive BM was recorded and compared with patients who underwent ABMT with noncontaminated BM grafts . RESULTS: In 239 BM grafts processed, the incidence of microbiological contamination was 26.4% (n = 63) . Fifty marrow grafts were contaminated by bacteria from the skin flora: coagulase-negative Staphylococcus (CNSC), Propionibacterium, and Corynebacterium species (79%) . Thirty-eight patients underwent ABMT (day 0) with cryopreserved culture-positive BM, and 32 patients were evaluable for microbiological cultures at thawing: in 10 of 32 BM grafts CNSC was found prior to reinfusion . Following ABMT, PB cultures revealed CNSC in 5 of 38 patients between days +4 and +12 . However, the late occurrence of positive PB cultures after BM reinfusion made a relationship between BM CNSC and PB CNSC unlikely . In 33 of 38 patients, no graft-contaminating bacteria were detected in PB . Comparison of the posttransplantation course of patients who received contaminated BM with that of patients grafted with noncontaminated BM showed no significant differences concerning time to engraftment, febrile days, and days on antibiotics . CONCLUSION: (1) Collection and/or ex vivo processing can result in microbiological contamination of BM grafts predominantly with bacteria from the skin flora, and (2) only CNSC can be cultured at thawing from previously contaminated/cryopreserved BM . Since patients undergoing ABMT usually receive oral antibiotics from beginning of the conditioning regimen which are active against CNSC, no further administration of antibiotics is recommended for the reinfusion of bacterially contaminated BM grafts. J Clin Microbiol, 1998 Mar, 36(3), 624 - 7 Corynebacterium riegelii sp . nov., an unusual species isolated from female patients with urinary tract infections; Funke G et al.; Four strains of an unknown coryneform bacterium were isolated in pure culture from females with urinary tract infections . Strong urease activity and the ability to slowly ferment maltose but not glucose were the most significant phenotypic features of this catalase-positive, nonmotile, nonlipophilic, rod-shaped bacterium which served to distinguish it from all other presently defined coryneform bacteria . Chemotaxonomic investigations demonstrated that the unknown bacterium belonged to the genus Corynebacterium . Comparative 16S rRNA gene sequence analysis revealed that the isolates were genealogically identical and represented a new subline within the genus Corynebacterium, for which the designation Corynebacterium riegelii sp . nov . is proposed . The type strain of Corynebacterium riegelii is CCUG 38180 (DSM 44326, CIP 105310). Ophthalmology, 1998 Mar, 105(3), 517 - 21 Nonulcerating bacterial keratitis associated with soft and rigid contact lens wear; McLeod SD et al.; OBJECTIVE: An unusual presentation of contact lens-related bacterial keratitis is that of epithelial nodular infiltrates and stromal inflammation without epithelial ulceration . The authors study the initial diagnosis, clinical features, causative organisms, and outcomes of corneal infections presenting in this manner . DESIGN: The study design was a 20-month retrospective chart review . PARTICIPANTS: Five patients with culture-proven bacterial keratitis who had predominantly nodular epithelial lesions were studied . RESULTS: Four infections were associated with soft contact lens wear and one with rigid lens wear . All patients had largely intact epithelium; typical gray-colored epithelial nodules, some with underlying anterior stromal haze; and diffuse, fine, cellular stromal inflammation . Two patients were referred with the tentative diagnosis of Acanthamoeba infection and two as contact lens-related sterile keratitis . Epithelial cultures from three cases yielded Serratia sp., one yielded Corynebacterium, and one Streptococcus pneumoniae . All responded to antibacterial medication; final corrected visual acuity in all cases was 20/30 or better . CONCLUSIONS: Bacterial infection associated with contact lens wear can be established within the corneal epithelium without initially producing an ulcer . A wide range of both gram-positive and gram-negative organisms can be involved . Early recognition and treatment appear to result in a favorable outcome. J Biol Chem, 1998 Feb 20, 273(8), 4329 - 37 The manganese-containing ribonucleotide reductase of Corynebacterium ammoniagenes is a class Ib enzyme; Fieschi F et al.; Ribonucleotide reductases (RNRs) are key enzymes in living cells that provide the precursors of DNA synthesis . The three characterized classes of RNRs differ by their metal cofactor and their stable organic radical . We have purified to near homogeneity the enzymatically active Mn-containing RNR of Corynebacterium ammoniagenes, previously claimed to represent a fourth RNR class . N-terminal and internal peptide sequence analyses clearly indicate that the C . ammoniagenes RNR is a class Ib enzyme . In parallel, we have cloned a 10-kilobase pair fragment from C . ammoniagenes genomic DNA, using primers specific for the known class Ib RNR . The cloned class Ib locus contains the nrdHIEF genes typical for class Ib RNR operon . The deduced amino acid sequences of the nrdE and nrdF genes matched the peptides from the active enzyme, demonstrating that C . ammoniagenes RNR is composed of R1E and R2F components typical of class Ib . We also show that the Mn-containing RNR has a specificity for the NrdH-redoxin and a response to allosteric effectors that are typical of class Ib RNRs . Electron paramagnetic resonance and atomic absorption analyses confirm the presence of Mn as a cofactor and show, for the first time, insignificant amounts of iron and cobalt found in the other classes of RNR . Our discovery that C . ammoniagenes RNR is a class Ib enzyme and possesses all the highly conserved amino acid side chains that are known to ligate two ferric ions in other class I RNRs evokes new, challenging questions about the control of the metal site specificity in RNR . The cloning of the entire NrdHIEF locus of C . ammoniagenes will facilitate further studies along these lines. Infection, 1998 Jan-Feb, 26(1), 36 - 8 Diversity of coryneforms found in infections following prosthetic joint insertion and open fractures; von Graevenitz A et al.; In a 5-year period, 73 coryneform isolates from prosthetic joint and open fracture infections in 60 patients treated in a hospital specialized in orthopedic surgery were speciated . The most frequent species were Corynebacterium amycolatum, Corynebacterium striatum, Corynebacterium diphtheriae biotype mitis, and Corynebacterium jeikeium . At least 14 isolates were deemed clinically significant as sole agents of infection. Exp Neurol, 1998 Feb, 149(2), 322 - 8 Entry of monocytes into the brain after injection of Corynebacterium parvum; Cheng L et al.; The receptiveness of the brain to monocyte infiltration was studied in rats that had been injected intracerebrally with Corynebacterium parvum . At 0-17 days after intracerebral injection and 18 h after intravenous injection of diI-labeled isogenous mononuclear cells, host rats were sacrificed and cells from the vicinity of the injection site and from the contralateral cerebral hemisphere were dissociated and analyzed by flow cytometry . In rats sacrificed 4-11 days postinjection of C . parvum, diI-labeled mononuclear cells were detected in cell preparations from the hemisphere ipsilateral and, to a lesser extent, contralateral to the injection site . No extravasation of cells from the blood to the brain was detected in rats injected intracerebrally with saline . By immunohistochemistry, many macrophages were detected in the hemisphere ipsilateral to injection of C . parvum . In additional experiments, the dissociated CNS cell population was labeled with OX-42 antibodies to the type 3 complement receptor, which is present on monocytes but not lymphocytes . Some cells in the brain were labeled with both diI and OX-42 and therefore were identified as monocytes that had entered the brain from the blood . In conclusion, monocytes can home to both sides of the brain after unilateral injection of a strong inflammatory agent but monocyte infiltration into the brain is delayed in comparison to monocyte inflammatory responses that have been reported in nonneural tissues. Rom J Virol, 1996 Jan-Dec, 47(1-4), 75 - 80 Immunomodulating and antiviral therapy in herpes zoster; Topciu V et al.; Two groups of patients with herpes zoster were followed up . The first group was subjected, beside a symptomatic therapy, to an immunological and antiviral treatment . The control group was treated only symptomatically . The immunological preparations used were: the immunostimulant SRE (Corynebacterium parvum), which stimulated the lymphocytes and macrophages, Moroxidin (Virustat-Paris) and Antiherpin (interferon inductor), which acted by blocking the virus replication . The preparations were indigenous and atoxic . A significant difference between the courses of disease in the two groups was observed, namely, the severity and duration of subjective and objective symptoms were more than double and followed by persistent neurological sequelae in the control group in comparison with the patients of the experimental group. J Dairy Sci, 1998 Jan, 81(1), 116 - 20 Efficacy of postmilking disinfection with benzyl alcohol versus lodophor in the prevention of new intramammary infections in lactating cows; Erskine RJ et al.; Five Michigan dairy herds participated in a split-herd study to compare the efficacy of two postmilking teat dips in the prevention of new intramammary infections (IMI) in lactating cows . Three hundred seventy cows were assigned to 4% benzyl alcohol, and 387 cows were assigned to 1% iodophor germicidal teat dip . The teat dips were applied by directly immersing the teats immediately after milking . Once a group was assigned to a teat dip, cows in that group maintained on that same teat dip throughout the trial . Total new IMI numbered 254 and 201 for cows treated with benzyl alcohol and iodophor germicidal teat dip, respectively . Staphylococcus spp . (52.0%), Staphylococcus aureus (20.1%), and Corynebacterium bovis (12.2%) were the predominant pathogens that caused new IMI in cows treated with benzyl alcohol . Staphylococcus spp., Staph . aureus, and C . bovis, respectively, were the pathogens responsible for 69.7, 12.4, and 4.5% of the new IMI in cows treated with iodophor . The incidences of new IMI caused by Staph . aureus (0.66 new IMI/100 milking quarters per mo), C . bovis (0.38 new IMI/100 milking quarters per mo), and all pathogens (3.15 new IMI/100 milking quarters per mo) were higher in cows treated with benzyl alcohol than in cows treated with iodophor (0.29, 0.11, and 2.35 new IMI/100 milking quarters per mo, respectively) . Incidence of new IMI did not differ between groups for other pathogens . One percent iodophor prevented new IMI caused by contagious pathogens more effectively than did benzyl alcohol. J Protein Chem, 1998 Jan, 17(1), 29 - 36 Extracellular poly(alpha-L-guluronate)lyase from Corynebacterium sp.: purification, characteristics, and conformational properties; Matsubara Y et al.; Extracellular alginate lyase was purified from the culture supernatant of Corynebacterium sp . isolated from the sewage of a sea tangle processing factory in order to elucidate the structure-function relationship of alginate lyase . The electrophoretically homogeneous enzyme was shown to have a molecular mass of 27 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and by gel filtration, with an isoelectric point of 7.3 . The molecular mass from amino acid analysis was 28.644 kDa . The optimal pH and temperature for the enzyme reaction were around 7.0 and 55 degrees C, respectively . Metal compounds such as MnCl2 and NiCl2 increased the enzyme activity . The enzyme was identified as the endolytic poly(alpha-L-guluronate)lyase, which was active on poly(alpha-L-1,4-guluronate) and caused a rapid decrease in the viscosity of alginate solution . Measurement of the far-UV circular dichroic spectrum of the enzyme molecule gave a spectrum with a deep trough at 215 nm accompanied by a shallow one at around 237 nm, and with a high peak at 197 nm and a much lower one at 230 nm . This spectrum was most likely to be that of the beta-form of the enzyme molecule and resembled poly(beta-D-mannuronate)lyase from Turbo cornutus (wreath shell) and poly(alpha-L-guluronate)lyase from Vibrio sp . (marine bacterium) . The near-UV circular dichroic spectrum was characteristic for aromatic amino acid residues . In the presence of 6 M urea, these spectra changed drastically in the near-UV and a little in the far-UV with the disappearance of the enzyme activity . Removal of the denaturant in the enzyme solution by dialysis restored both the activity and inherent circular dichroic spectra . The beta-sheets observed in alginate lyases as the major ordered structure seem to be a common conformation for the lyases. Diagn Microbiol Infect Dis, 1998 Jan, 30(1), 7 - 15 Prospective study of catalase-positive coryneform organisms in clinical specimens: identification, clinical relevance, and antibiotic susceptibility; Lagrou K et al.; During a 6-month period, all clinical isolates of catalase-positive coryneform organisms, which were isolated during the routine processing of clinical specimens, were characterized in the laboratory of the 1800-bed University Hospital of Leuven . The distribution of the species in the corynebacteria was: Corynebacterium amycolatum 70 (53%), Corynebacterium jeikeium 16 (12%), Corynebacterium striatum 11 (8%), Corynebacterium afermentans 10 (7%), Corynebacterium minutissimum 9 (6%), CDC coryneform group G 4 (3%), Corynebacterium urealyticum 4 (3%), Corynebacterium glucuronolyticum 1 (0.7%), and Corynebacterium xerosis 1 (0.7%) . Of the 150 isolates, 37 (25%) were considered to be infection related and the remaining 113 (75%) were of questionable clinical significance . Susceptibility of the corynebacteria to 12 antibiotics active against Gram-positive organisms was evaluated . C . amycolatum, C . jeikeium, and C . urealyticum were multiresistant, but all isolates were susceptible to teicoplanin and vancomycin . Most of the C . amycolatum strains, and all strains of C . jeikeium and C . striatum, were susceptible to the vibrocidal compound O/129. Appl Microbiol Biotechnol, 1998 Jan, 49(1), 24 - 30 Improved L-lysine yield with Corynebacterium glutamicum: use of dapA resulting in increased flux combined with growth limitation; Eggeling L et al.; The amino acid L-lysine is produced on a large scale using mutants of Corynebacterium glutamicum . However, as yet recombinant DNA techniques have not succeed in improving strains selected for decades by classic mutagenesis for high productivity . We here report that seven biosynthetic enzymes were assayed and oversynthesis of the dihydrodipicolinate synthase resulted in an increase of lysine accumulation from 220 mM to 270 mM . The synthase, encoded by dapA, is located at the branch point of metabolite distribution to either lysine or threonine and competes with homoserine dehydrogenase for the common substrate aspartate semialdehyde . When graded dapA expression was used, as well as quantification of enzyme activities, intracellular metabolite concentrations and flux rates, a global response of the carbon metabolism to the synthase activity became apparent: the increased flux towards lysine was accompanied by a decreased flux towards threonine . This resulted in a decreased growth rate, but increased intracellular levels of pyruvate-derived valine and alanine . Therefore, modulating the flux at the branch point results in an intrinsically introduced growth limitation with increased intracellular precursor supply for lysine synthesis . This does not only achieve an increase in lysine yield but this example of an intracellularly introduced growth limitation is proposed as a new general means of increasing flux for industrial metabolite over-production. Antibiot Khimioter, 1997, 42(12), 16 - 8 {Susceptibility of Corynebacteria isolated in St Petersburg to antibacterial drugs}; Gladin DP et al.; Susceptibility of 150 Corynebacterium isolates (91 strains of C.pseudodiphtheriticum and 59 strains of the ANF group corynebacteria) to 21 antibacterial drugs was determined by the method of serial dilutions in a solid medium . It was shown that the MIC of the drugs for the diphtheroids was within the ranges of < 0.015 to > 32.0 micrograms/ml . 66 per cent of the Corynebacterium strains circulating in St . Petersburg was resistant at least to 1 antibacterial drug . The Corynebacterium isolates with moderate resistance to erythromycin and lincomycin (57.3 per cent) and resistant to trimethoprime (16.7 per cent) were the most frequent . 8.0 per cent of the diphtheroids was resistant at least to 4 antibacterial drugs . No significant difference in the susceptibility of the ANF group corynebacteria and C.pseudodiphtheriticum to the drugs was observed . Gentamicin, rifampicin, tetracycline and doxycycline showed high activity against the corynebacteria at present circulating in St . Petersburg . When antibacterial therapy of the infection due to corynebacteria fails it is necessary to estimate antibioticograms of Corynebacterium pure cultures. Mikrobiol Z, 1997 Sep-Oct, 59(5), 22 - 8 {The serological properties of saprophytic corynebacteria studied by immunoenzyme analysis}; Mikhal'skii LA et al.; The degree of serological similarity of saprophytic corynebacteria have been studied using immunoassay ELISA analysis, that is seven collection strains, belonging to Corynebacterium glutamicum (3 strains), C . ammoniagenes (1 strain), C . vitarumen (1 strain), C . variabilis (2 strains) and three industrial strains-lysine producers . Intact and heated bacteria cells have been used as antigens . It has been shown that industrial strain C . glutamicum 22L and collection strains C . glutamicum IMV AC-715, IMV AC-714, IMV AC-733 have the highest degree of serological relationship . C . vitarumen IMV AC-718, C variabilis IMV AC-716 as well as Corynebacterium sp . E531 and VNIIgenetics 90 are close to them according to their serological properties . C . ammoniagenes IMV AC-732 and C . variabilis IMV AC-717 strains have the lowest degree of similarity with other saprophytic corynebacteria which have been studied. J Biol Chem, 1998 Jan 30, 273(5), 2567 - 74 Osmo-sensing by N- and C-terminal extensions of the glycine betaine uptake system BetP of Corynebacterium glutamicum; Peter H et al.; The major uptake carrier for the compatible solute glycine betaine in Corynebacterium glutamicum is the secondary transport system BetP . It is effectively regulated by the external osmolality both on the level of expression and of activity . BetP carries highly charged domains both at the N and at the C terminus . We investigated the role of these extensions in the regulatory response to hyperosmotic stress . Mutants of the betP gene coding for proteins with truncated N- and C-terminal extensions were expressed in the C . glutamicum betP deletion strain DHP1 and were functionally characterized with respect to regulation of activity . The optimum of activation at 1.3 osmol/kg in wild type was shifted in the recombinant strains to about 2.6 osmol/kg in mutants with deletions in the N-terminal part . Deletions in the C-terminal domain resulted in a complete loss of regulation . The altered response to changes in osmolality led to severe consequences in the cellular adaption to hyperosmotic stress . Whereas in the wild type, the steady state level of glycine betaine accumulation is maintained by activity regulation of the BetP system itself, in the mutant with BetP proteins carrying truncations in the C-terminal domain, the observed steady state betaine accumulation was found to be due to a kinetic balance of unregulated glycine betaine uptake by the modifed BetP and efflux via the mechanosensitive efflux channel for compatible solutes at the same time. Br J Cancer, 1998, 77(3), 426 - 33 Persistent induction of nitric oxide synthase in tumours from mice treated with the anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid; Moilanen E et al.; An anti-tumour agent 5,6-dimethylxanthenone-4-acetic acid (5,6-MeXAA) induced nitric oxide synthase (NOS) in the tumour, spleen, thymus and small intestine, but not in the lung, liver, kidney, heart or skeletal muscle in B6D2F1 mice bearing subcutaneous colon 38 tumours . This pattern of induction is distinct from that caused by agents such as endotoxin, muramyl dipeptide or Corynebacterium parvum . The induction of NOS (iNOS) in the tumour was more persistent (maximal at 3 days) than in other tissues (maximal at 12 h) . Immunohistochemical staining suggested that iNOS was located in macrophages and endothelial cells within and around the tumour . Treatment with 5,6-MeXAA also caused substantial increases in plasma nitrite and nitrate (NOx) concentrations that peaked at 8-12 h after 5,6-MeXAA . The increase in plasma NOx was prevented by a NOS inhibitor N-iminoethyl-L-ornithine (L-NIO), indicating that it was due to enhanced production of NO . Tumour-bearing mice were more responsive than controls to 5,6-MeXAA both in their plasma NOx increase and in their lower maximally tolerated dose . L-NIO was unable to prevent the complete tumour necrosis and regression caused by 5,6-MeXAA at a dose that substantially inhibited the increase of plasma NOx . In conclusion, the experimental anti-tumour agent 5,6-MeXAA induced NO synthesis in tumour-associated macrophages and in immunologically active tissues in parallel with its effects on tumour growth . The experiments with a non-selective NOS inhibitor L-NIO, however, suggest that NO is not a significant component in the mechanism of the anti-tumour action of 5,6-MeXAA in this particular model. Epidemiol Mikrobiol Imunol, 1997 Sep, 46(3), 87 - 98 {Hemolysis and hemolytic interactions of bacteria}; Skalka B et al.; The review contains data on haemolytically active bacteria important in medical microbiology, human or veterinary . Attention is paid to haemolytically active bacterial metabolites, especially those which produce synergistic or antagonistic haemolytic reactions above all with staphylococcal haemolysins alpha, beta and delta, with corynebacterial phospholipase D, with rhodococcal equi factor and with the haemolysin of Dermatophilus congolensis . The general usage of the term "CAMP factor" for all manifestations of haemolytic interactions is criticized . Descriptions of haemolytic activities are supplemented by a synoptical table and most of them are documented by photographs. Acta Ophthalmol Scand, 1997 Oct, 75(5), 592 - 4 Experience with a broth culture technique for diagnosis of bacterial keratitis; Schonheyder HC et al.; PURPOSE: To determine if a broth culture technique is a practical means for bacteriological investigation of keratitis . MATERIAL AND METHODS: Twenty-seven eyes of 27 patients with a clinical diagnosis of bacterial keratitis were included in a prospective and non-comparative study at a Danish referral hospital . A corneal scrape was inoculated directly into broth medium which was transferred to the diagnostic laboratory for incubation and subculture . RESULTS: Culture was negative in 4 patients, and 19 of the remaining 23 patients had a pure growth of either Pseudomonas aeruginosa (n = 8), Staphylococcus aureus (n = 2), Streptococcus pneumoniae (n = 2), Haemophilus influenzae biotype III (n = 1), Moraxella species (n = 1), Corynebacterium species (n = 1), or coagulase-negative staphylococci (n = 4) . In 4 patients there was a mixed gram-positive growth . There was no association between microbiological findings and previous topical antibiotic therapy . Contamination and lack of quantitative assessment of growth proved not to be a problem . CONCLUSIONS: By broth culture technique we identified a definite pathogen (P . aeruginosa, S . aureus or S . pneumoniae) in 44% of patients (95% binomial confidence limits: 25-65%) . The technique may replace the standard technique of direct plate culture under circumstances where it is difficult to keep a supply of fresh media or transport inoculated plates. J Chromatogr A, 1998 Jan 9, 793(1), 214 - 9 Determination of 4-nitrocatechol in biodegradation samples by gas chromatography-mass spectrometry; Zdrahal Z; 4-Nitrocatechol was identified as a product of transformation of 4-nitrophenol by bacterial strain Corynebacterium sp.8/3 using direct acetylation of biodegradation samples by acetic anhydride followed by GC-MS analysis . The identity of 4-nitrocatechol, in the form of diacetate, was confirmed by electron-impact spectra and spectra recorded under chemical ionization conditions (positive and negative modes) . Negative-ion chemical ionization was used for quantification of 4-nitrocatechol in biodegradation samples in a concentration range of 1-25 mg/l. Zh Mikrobiol Epidemiol Immunobiol, 1997 Nov-Dec, (6), 61 - 4 {The characteristics of the lymphocyte subpopulation composition and its clinical significance in bacterial carriers of toxigenic Corynebacterium diphtheriae}; Iushchuk ND et al.; In 43 carriers of toxigenic C . diphtheriae the antigenic picture of lymphocyte membranes was studied by the method of flow cytofluorometry . The study revealed that at the time of the hospitalization of the carriers a decrease in the number of CD21+ and HDL-DR+ lymphocytes was a prognostically important criterion for the formation of carrier state, while a decrease in the content of natural killers (CD16+) was characteristic of prolonged forms of carrier state . The necessity of the study of cellular immunity factors in carriers at the time of their hospitalization and the expediency of the inclusion of immunocorrective therapy into the individual schemes of their treatment were substantiated. J Biol Chem, 1998 Jan 9, 273(2), 837 - 41 Expression and characterization of a heme oxygenase (Hmu O) from Corynebacterium diphtheriae . Iron acquisition requires oxidative cleavage of the heme macrocycle; Wilks A et al.; A full-length heme oxygenase gene from the pathogenic bacterium Corynebacterium diphtheriae has been subcloned and expressed in Escherichia coli . The enzyme is expressed at high levels as a soluble catalytically active protein that results in the accumulation of biliverdin within the E . coli cells . The purified heme oxygenase forms a 1:1 complex with heme (Kd = 2.5 +/- 1 microM) and has hemeprotein spectra similar to those previously reported for the purified eukaryotic heme oxygenases . In the presence of an E . coli NADPH-dependent reductase isolated during the purification of Hmu O, the heme-Hmu O complex is catalytically turned over to yield biliverdin IXalpha and carbon monoxide . A number of redox partners were investigated for their ability to reconstitute Hmu O activity in vitro . Of these the most efficient appeared to be the recombinant NADH-dependent putidaredoxin/putidaredoxin reductase from Pseudomonas putida . As with the E . coli NADPH-dependent reductase the final products of the reaction were biliverdin IXalpha and carbon monoxide . This is the first bacterial heme oxygenase to be described to date . The close relationship between iron acquisition and pathogenesis suggests that the release of iron from heme by heme oxygenase may play a crucial role in the pathogenicity of C . diphtheriae. J Clin Microbiol, 1998 Jan, 36(1), 207 - 10 Antitoxin-in-membrane and antitoxin-in-well assays for detection of toxigenic Corynebacterium diphtheriae; Reinhardt DJ et al.; The Elek culture plate precipitin test is routinely used for the detection of exotoxin from toxigenic strains of Corynebacterium diphtheriae . Recently, the World Health Organization standardized this test to ensure accuracy, reliability, and reproducibility . In this study, we further modified the standard Elek test by using the antitoxin-in-membrane (AIM) and antitoxin-in-well (AIW) approaches . In the AIM tests, each strain was stabbed and streaked backwards and away from a point approximately 7 mm from the edge of a sterile cellulose acetate-cellulose nitrate filter membrane disk (pore size, 0.45 microm; diameter, 25 mm) containing 25 IU of diphtheria antitoxin . For AIW tests, a central well (diameter, 5 mm) containing 9 microl of antitoxin (4.5 IU) was surrounded by eight equidistant stab-streaks of each strain placed 10 mm from the well . In both methods, precipitin bands of identity typically were noted after 24- and 48-h incubations at 37 degrees C . Both toxigenic and weak toxigenic strains gave clear and reproducible results . Compared with the standard Elek test, the AIM and AIW tests each use 50% less medium and 75 and 87% less antitoxin, respectively . AIM has the potential to test up to 14 isolates and AIW has the potential to test up to 24 isolates on the same plate . Furthermore, clearer positives were noted with weak toxigenic strains . In a blinded test of 209 verified C . diphtheriae isolates, a 99.5% agreement with the standard Elek test was obtained overall . Both modifications conserve reagents and medium, permit the simultaneous testing of a larger number of strains, and may be particularly suitable for reference laboratories or hospitals involved in diphtheria epidemic settings. Appl Microbiol Biotechnol, 1997 Dec, 48(6), 693 - 8 A novel process of inosine 5'-monophosphate production using overexpressed guanosine/inosine kinase; Mori H et al.; A novel process for producing inosine 5'-monophosphate (5'-IMP) has been demonstrated . The process consists of two sequential bioreactions; the first is a fermentation of inosine by a mutant of Corynebacterium ammoniagenes, and the second is a unique phosphorylating reaction of inosine by guanosine/inosine kinase (GIKase) . GIKase was produced by an Escherichia coli recombinant strain, MC1000(pIK75), which overexpressed the enzyme up to 50% of the total cellular protein . The overproducing plasmid, pIK75, which was randomly screened out from deletion plasmids with various lengths of intermediate sequence between the E . coli trpL Shine-Dalgarno sequence, derived from the vector plasmid, and the start codon of the GIKase structural gene . In pIK75, the start ATG was placed 16 bp downstream of the trpL Shine-Dalgarno sequence under the control of the E . coli trp promoter . Fermentation of inosine and its phosphorylation were sequentially performed in a 5-1 jar fermenter . At the end of inosine fermentation by C . ammoniagenes KY13761, culture broth of MC1000(pIK75) was mixed with that of KY13761 to start the phosphorylating reaction . Inosine in the reaction mixture was stoichiometrically phosphorylated, and 91 mM 5'-IMP accumulated in a 12-h reaction . This new biological process has advantages over traditional methods for producing 5'-IMP. Infect Immun, 1998 Feb, 66(2), 474 - 9 Vaccine potential of attenuated mutants of Corynebacterium pseudotuberculosis in sheep; Simmons CP et al.; Corynebacterium pseudotuberculosis, a gram-positive facultative intracellular bacterial pathogen, is the etiological agent of the economically important disease caseous lymphadenitis (CLA) in both sheep and goats . Attenuated mutants of C . pseudotuberculosis have the potential to act as novel vaccines against CLA and as veterinary vaccine vectors . In this report, we have assessed the virulence of both aroQ and pld mutants of C . pseudotuberculosis in sheep and concurrently their capacity to act as vaccines against homologous challenge . The results suggest that aroQ mutants of C . pseudotuberculosis are attenuated with regard to both lymph node persistence and vaccination site reactogenicity . Immunologically, aroQ mutants failed to elicit detectable specific gamma interferon (IFN-gamma)-secreting lymphocytes and induced low levels of antibodies to C . pseudotuberculosis culture supernatant antigens . Following subcutaneous vaccination, the immune responses induced by aroQ mutants did not protect sheep from infection with the wild-type strain but did appear to reduce the clinical severity of disease resulting from challenge . Conversely, an attenuated C . pseudotuberculosis strain expressing an enzymatically inactive phospholipase D exotoxin, when used as a vaccine, elicited a protective immune response . Protection appeared to correlate with in vivo persistence of the vaccine strain, the induction of IFN-gamma-secreting lymphocytes, and relatively high levels of antibodies to culture supernatant antigens . The results suggest that aroQ mutants of C . pseudotuberculosis may be overly attenuated for use as a CLA vaccines or as vaccine vectors. Ann Pharm Fr, 1997, 55(6), 262 - 8 Isolation, identification and antimicrobial activity of ombuoside from Stevia triflora; Amaro-Luis JM et al.; From aerial parts of Stevia triflora DC the flavonol glycoside ombuoside (7,4'-di-O-methylquercetin-3-O-beta-rutinoside) has been isolated and identified on the basis of spectral data . Ombuoside and the synthetic derivatives octa-acetylombuoside, ombuine and retusine were tested for antimicrobial activity against several strains of Gram-positive and Gram-negative bacteria and the yeast Candida albicans, using the agar diffusion method . The flavonol glycoside ombuoside and the respective aglycone ombuine, both exhibited moderated activity against Corynebacterium diphtheria, Staphylococcus aureus, Escherichia coli and Candida albicans . To a lesser degree, octaacetylombuoside and retusine showed activity against the Gram-positive bacteria C . diphtheria and S . aureus, but proved to be inactive against Gram-negative bacteria and Candida albicans . These results indicate that the presence of free hydroxyl groups, either alcoholic or phenolic, is an important chemical feature for the expression of flavonol antimicrobial activity . It is worth noting that this is the first study reported on the antibacterial and antifungal activity of these substances. J Med Microbiol, 1998 Jan, 47(1), 79 - 83 Evaluation of a new selective medium for the isolation of Corynebacterium urealyticum; Zapardiel J et al.; A new selective medium (CBU agar) was compared with blood agar (BA) medium for primary isolation of Corynebacterium urealyticum from urine and skin samples of hospitalised patients . Overall, the CBU agar detected C . urealyticum in 14 (4.6%) of 302 urine samples and the BA medium detected the organism in four (1.3%), but most cultures which were positive only on CBU agar had < 10(4) cfu/ml . Six strains of C . urealyticum were isolated from 60 skin samples with CBU agar, whereas none was detected with BA . Although most skin samples had heavy inocula, the selective agar facilitated the recognition of low colony counts (< or = 10 cfu/plate) of C . urealyticum by reducing the growth of competing flora . Challenge of the selective medium with reference and clinical strains showed that CBU agar was inhibitory for gram-negative bacteria and reduced the gram-positive flora, allowing the growth of C . urealyticum strains . The new selective medium appears to be a useful epidemiological tool to study urinary and skin colonisation by C . urealyticum. Eur J Clin Microbiol Infect Dis, 1997 Nov, 16(11), 816 - 20 Fatal respiratory tract diphtheria apparently caused by nontoxigenic strains of Corynebacterium diphtheriae; Rakhmanova AG et al.; A major diphtheria epidemic affecting the whole population of St . Petersburg started in 1990 . During the period of 1991 to 1995, 4600 patients with clinical respiratory tract diphtheria were treated in Botkin's Hospital . From 112 (2.4%) of these patients only a nontoxigenic strain of Corynebacterium diphtheriae was isolated . Three patients with this strain who were suffering from clinical disease consistent with classical toxic diphtheria died . All had myocarditis, two had asphyxia due to membrane formation in the lower respiratory tract, and one had severe polyneuritis . In two patients the causative agent was of the biotype mitis and in the third intermedius, whereas the prevailing epidemic strain was of the biotype gravis . As the clinical presentation of the disease in the three patients who died was typical of toxic diphtheria, it is considered likely that the immunodiffusion test for toxin production in vitro may fail to detect strains of Corynebacterium diphtheriae producing toxin in vivo. Microb Drug Resist, 1997 Winter, 3(4), 345 - 50 Drug extrusion in Corynebacterium glutamicum; Kaidoh K et al.; We selected a mutant of Corynebacterium glutamicum, EBR, which can grow in a medium containing cytotoxic ethidium bromide (EtBr) at a high concentration of 100 microM . The resistance to EtBr in the mutant was reversed by 2 microM reserpine, a potent inhibitor of mammalian p-glycoprotein and bacterial multidrug resistance (MDR) transporter, whereas reserpine alone had a minimal effect on cell growth . The mutant showed a much higher efflux rate of EtBr than wild-type cells, and the efflux was completely inhibited by 2 microM reserpine . In addition to reserpine, structurally unrelated chemicals such as quinidine, trifluorperazine, tetraphenylarsonium chloride, chlorpromazine and quinine inhibit the EtBr efflux, revealing that the putative efflux system(s) can recognize a variety of chemicals . The efflux activity was correlated with the membrane potential but not the intracellular ATP contents . We, therefore, concluded that the EtBr resistance may be involved by proton-motive-force driven multidrug efflux system(s). Can Vet J, 1998 Jan, 39(1), 33 - 8 Clinical mastitis in dairy cattle in Ontario: frequency of occurrence and bacteriological isolates; Sargeant JM et al.; The objective of this study was to describe the frequency of occurrence of clinical mastitis in dairy herds in Ontario . The study group consisted of 65 dairy farms involved in a 2-year observational study, which included recording all clinical mastitis cases and milk sampling of quarters with clinical mastitis . Lactational incidence risks of 9.8% for abnormal milk only, 8.2% for abnormal milk with a hard or swollen udder, and 4.4% for abnormal milk plus systemic signs of illness related to mastitis were calculated for 2840 cows and heifers . Overall, 19.8% of cows experienced one or more cases of clinical mastitis during location . Teat injuries occurred in 2.1% of lactations . Standard bacteriology was performed on pretreatment milk samples from 834 cows with clinical mastitis . The bacteria isolated were Staphylococcus aureus (6.7%), Streptococcus agalactiae (0.7%), other Streptococcus spp . (14.1%), coliforms (17.2%), gram-positive bacilli (5.5%), Corynebacterium bovis (1.7%), and other Staphylococcus spp . (28.7%) . There was no growth in 17.7% of samples, and 8.3% of samples were contaminated . Clinical mastitis is a common disease in dairy cows in Ontario; approximately 1 in 5 cow lactations have at lease one episode of clinical mastitis . There is, however, considerable variation in the incidence of clinical mastitis among farms . The majority of 1st cases of clinical mastitis occur early in lactation, and the risk of clinical mastitis increases with increasing parity . Environmental, contagious, and minor pathogens were all associated with cases of clinical mastitis. Can J Vet Res, 1998 Jan, 62(1), 38 - 43 The incidence of caseous lymphadenitis in Alberta sheep and assessment of impact by vaccination with commercial and experimental vaccines; Stanford K et al.; In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass condemnation . In this study, serologic results confirmed a high (50-94%) incidence of exposure to Corynebacterium pseudotuberculosis, the causative agent of CLA, in mature, unvaccinated sheep in southern Alberta . To assess the efficacy and impact of vaccination with 2 commercial (Glanvac-6 and Case-Vac) and 1 experimental (WC+ MDP-GDP) CLA vaccines, a series of 3 field trials in 3249 ewes and lambs was conducted in affected flocks from 1992-1996 . Efficacy was assessed from the serological response to vaccination, prevalence and size of injection site reactions by treatment, and the incidence of CLA abscesses . Overall, agglutinating antibody titres to C . pseudotuberculosis in lambs vaccinated with WC+MDP-GDP and Case-Vac remained significantly elevated above nonvaccinated control lambs for the 12 mo period after the initial vaccination . Lambs vaccinated with the WC/MDP-GDP maintained higher titres (P < 0.06) than those vaccinated with Case-Vac for the period from 6 to 12 mo after vaccination . Agglutinating antibody titres for lambs vaccinated with Glanvac did not differ from those of controls at any point during the 12 mo period after vaccination . The number of injection site reactions was elevated in lambs vaccinated with Glanvac as compared to those vaccinated with WC+MDP-GDP but the size of injection site reactions did not significantly differ . Sheep vaccinated with WC+ MDP-GDP also had a reduced incidence of putative CLA abscesses, although confirmation of the presence of C . pseudotuberculosis was only successful in a small number of instances. Antonie Van Leeuwenhoek, 1997 Nov, 72(4), 291 - 7 Ultrastructure of the Corynebacterium glutamicum cell wall; Marienfeld S et al.; The cell wall structure of the Gram-positive Corynebacterium glutamicum was evaluated by electron microscopy of thin sections after freeze-substitution and conventional fixation with glutaraldehyde . For the cell wall an overall thickness of approximately 32 nm was determined, with 8.5 nm corresponding to an outer layer, 6.5 nm to an electron translucent region (ETR) as found in mycobacteria and 17 nm to the peptidoglycan . Knob-like surface structures previously observed in freeze-fracture experiments were detected when cells were conventionally processed with a fixation using glutaraldehyde . By mild treatment with detergents approximately 20 proteins were extracted from the cell wall . From seven of these N-terminal amino acid sequences were determined. Cornea, 1998 Jan, 17(1), 57 - 61 Antimicrobial effect of ciprofloxacin, povidone-iodine, and gentamicin in the decontamination of human donor globes; Gopinathan U et al.; PURPOSE: Clinical research addressing the issue of donor globe decontamination is yet to establish convincing data for the optimal choice of an antimicrobial agent . METHODS: In a donor-globe decontamination study, the antimicrobial effectiveness of a fluoroquinolone antibiotic (ciprofloxacin, 0.3%) was evaluated for the first time and compared with povidone-iodine (P-I, 5%) and gentamicin (0.3%) . RESULTS: Ciprofloxacin and gentamicin were found to be less effective than P-I (p < 0.05) in converting culture-positive donor globes to culture negative . In eliminating coagulase-negative staphylococci that predominated the bacterial spectrum, again P-I scored better than ciprofloxacin (p = 0.003) and gentamicin (p = 0.006) . Overall, P-I performed better than the other two in the 3-min decontamination procedure . Decontamination was carried out with the same agent for 15 min to assess the effect of duration of decontamination on the antimicrobial activity of P-I . With time, there was no significant increase in the antimicrobial efficacy of the agent except for Corynebacterium species . CONCLUSION: P-I continues to be the preferred agent for decontaminating donor globes . Whereas a contact of 3-min duration between P-I and donor globe remains satisfactory in decontamination procedures, corneal tolerance of this procedure needs investigation. J Dairy Sci, 1997 Dec, 80(12), 3219 - 26 Influence of parity and stage of lactation on the somatic cell count in bacteriologically negative dairy cows; Laevens H et al.; This study examines the influence of parity, stage of lactation, and single isolations (i.e., the isolation of a microorganism that could not be reisolated in the same quarter in the next sampling) of staphylococci other than Staphylococcus aureus (coagulase-negative staphylococci), Corynebacterium bovis, or esculin-positive cocci other than Streptococcus uberis (referred to as esculin-positive cocci throughout) on the monthly log(e)-transformed somatic cell count (SCC) for 180 first, second, and third parity cows that were observed over a whole lactation . Repeated measures ANOVA was used to analyze the data . No significant effect was found for the infection variable . However, the results indicated that even single isolations of coagulase-negative staphylocci, C . bovis, or esculin-positive cocci resulted in a numerical or statistically significant increase in SCC . Least squares mean SCC (log(e)-transformed) for bacterio-logically negative cows and cows with single isolations of coagulase-negative staphylococci, C . bovis, or esculin-positive cocci were 3.90, 3.97, 4.08, and 4.17, respectively . Significant effects of parity, stage of lactation, and the interaction of parity and stage of lactation could not be found when only bacteriologically negative cows were considered . Least squares mean SCC for first, second, and third parity cows were 3.80, 3.93, and 3.97, respectively . However, the effects of parity, stage of lactation, and the interaction of parity and stage of lactation were significant when all 180 cows were included . Therefore, these effects must be due to factors that were present in the infected groups. Arch Biochem Biophys, 1997 Dec 15, 348(2), 262 - 7 The monovalent cation requirement of rabbit muscle pyruvate kinase is eliminated by substitution of lysine for glutamate 117; Laughlin LT et al.; The crystal structure of rabbit muscle pyruvate kinase complexed with Mn2+, K+, and pyruvate revealed a binding site of K+ {T . M . Larsen, L . T . Laughlin, H . M . Holden, I . Rayment, and G . H . Reed (1994) Biochemistry 33, 6301-6309} . Sequence comparisons of rabbit muscle pyruvate kinase and pyruvate kinases from Corynebacterium glutamicum and Escherichia coli, which do not exhibit a requirement for activation by monovalent cations, indicate that the only substitutions in the K+ binding site are conservative . Glu 117 in the rabbit muscle enzyme, which is close to the K+ site, is, however, replaced by Lys in these two bacterial pyruvate kinases . The proximity of Glu 117 to K+ in the structure of the rabbit enzyme and conservation of the binding site in the bacterial enzymes which lack a dependence on monovalent cations suggested that a protonated epsilon-amino group of Lys 117 in these bacterial enzymes may provide an "internal monovalent cation." Site-specific mutant forms of the rabbit enzyme corresponding to E117K, E117A, E117D, and E117K/K114Q pyruvate kinase were examined to test this hypothesis . The E117K pyruvate kinase exhibits 12% of the activity of the fully activated wild-type enzyme but is > 200-fold more active than the wild-type enzyme in the absence of activating monovalent cations . Moreover, the activity of E117K pyruvate kinase exhibits no stimulation by monovalent cations in the assay mixtures . Both E117A and E117D pyruvate kinases retain activation by monovalent cations but have reduced activities relative to wild type . The results are consistent with the hypothesis that pyruvate kinases that do not require activation by monovalent cations supply an internal monovalent cation in the form of a protonated epsilon-amino group of Lys . The results also support the assignment of the monovalent cation in the active site of pyruvate kinase. Gene, 1997 Dec 12, 203(2), 95 - 101 The Corynebacterium glutamicum cglIM gene encoding a 5-cytosine methyltransferase enzyme confers a specific DNA methylation pattern in an McrBC-deficient Escherichia coli strain; Schafer A et al.; The cglIM gene of the coryneform soil bacterium Corynebacterium glutamicum ATCC 13032 has been cloned and characterized . The coding region comprises 1092 nucleotides and specifies a protein of 363 amino acid residues with a deduced Mr of 40700 . The amino acid sequence showed striking similarities to methyltransferase enzymes generating 5-methylcytosine residues, especially to M x NgoVII from Neisseria gonorrhoeae recognizing the sequence GCSGC . The cglIM gene is organized in an unusual operon which contains, in addition, two genes encoding stress-sensitive restriction enzymes . Using PCR techniques the entire gene including the promoter region was amplified from the wild-type chromosome and cloned in Escherichia coli . Expression of the cglIM gene in E . coli under the control of its own promoter conferred the C . glutamicum-specific methylation pattern to co-resident shuttle plasmids and led to a 260-fold increase in the transformation rate of C . glutamicum . In addition, the methylation pattern produced by this methyltransferase enzyme is responsible for the sensitivity of DNA from C . glutamicum to the modified cytosine restriction (Mcr) system of E . coli. Biochemistry, 1997 Dec 9, 36(49), 15201 - 7 Oligomerization of a 45 kilodalton fragment of diphtheria toxin at pH 5.0 to a molecule of 20-24 subunits; Bell CE et al.; Diphtheria toxin (DT) is a 58 kDa protein, secreted by lysogenic strains of Corynebacterium diphtheriae, that causes the disease diphtheria in humans . The catalytic (C) domain of DT kills host cells by gaining entry into the cytoplasm and inhibiting protein synthesis . The translocation of the C domain across the endosomal membrane and into the cytoplasm of a host cell is mediated by the translocation (T) domain of DT . This process is triggered by acidification from pH approximately 7 to pH approximately 5 within the endosome . Here we show that crm45 (cross-reacting material of 45 kDa), a 45 kDa deletion mutant of DT which contains the C and T domains but lacks the C-terminal receptor-binding (R) domain, undergoes a transition from a monomer to a large oligomer upon acidification from pH 7.0 to pH 5.0 . Dynamic light scattering analysis of crm45 at pH 5.0 results in a polydispersity value of only 8-17%, suggesting that the oligomer is uniformly sized . Using analytical ultracentrifugation, measurements of the sedimentation rate and diffusion coefficient of crm45 at pH 5.0 result in a molecular mass determination of 890 +/- 40 kDa (20 +/- 1 subunits) for the oligomer . Equilibrium sedimentation data on crm45 at pH 5.0 are best fit by a single species with a mass of 1000 +/- 50 kDa (24 +/- 1 subunits) . These results reveal the pH-dependent formation of a uniformly sized, 20-24 subunit oligomer of the C and T domains of DT, in solution . Because the oligomer of crm45 forms at the pH of the acidified endosome, it could be relevant to the translocation of the C domain of DT across the endosomal membrane and into the cytoplasm of host cells . The possible relevance of this oligomer of crm45 to the membrane translocation of the C domain of DT correlates with earlier kinetic studies of DT intoxication of Vero cells, which inferred the transfer of approximately 20 C domains of DT to the cytoplasm of host cells, in a single event. Stomatologiia (Mosk), 1997, 76(5), 4 - 8 {A trial of the use of rulid, sumamed and makropen in the combined treatment of generalized periodontitis at a stage of exacerbation}; Tsarev VN et al.; In vitro study of the antibacterial activity of macrolide antibiotics azitromycin (sumamed), midicamycin (macropen), roxitromycin (rulide), and erythromycin demonstrated their high activity towards clinical strains of bacteroids, fusobacteria, peptostreptococci, streptococci, and corynebacteria . These antibiotics were effective in the treatment of 62 adult patients with severe and moderate generalized periodontitis . Rulide and sumamed were the most effective, macropen and erythromycin were inferior to them. Rev Argent Microbiol, 1997 Jul-Sep, 29(3), 122 - 30 {Dynamics of bacteria from the phyllosphere and leaves of soy (Glycine max L . Merrill) in field conditions}; Salerno CM et al.; The surfaces of aerial plant provide a habitat for epiphitic microorganisms many of which are capable of influencing the growth of pathogens . In this study, fluctuation of bacterial population occurring in different growth stages of soybean leaves (Glycine max L Merrill) was examined using the dilution plate method . Three culture media were applied; invariably, the numbers of bacteria increased with increasing plant age . The total number of bacteria living on the detached leaves were ten times higher than leaves surfaces of soybean Asgrow 3127 (Group III) . One hundred and seventy-five heterotrophic bacteria were obtained from both the phyllosphere and litter of the plant . Of these, fifty-one microorganisms (29%) were classified as members of the genus Bacillus; the remainder were mainly members of the irregular, nonsporing, Gram positive rods (coryneform bacteria) . Approximately 52% of the total coryneform bacteria isolated was found to belong to the following two clusters: Arthrobacter and Corynebacterium . There were few Pseudomonas strains and 75% of the total isolated bacteria were able to grow in N-poor culture medium. Br J Dermatol, 1997 Oct, 137(4), 623 - 5 Breast abscess due to Corynebacterium striatum; Stone N et al.; Corynebacterium striatum, a normal constituent of the skin flora, is rarely pathogenic . Previous reports of infection are few, and are mainly confined to immunosuppressed patients or those with indwelling prosthetic devices . We report a case in which the organism caused a recurrent breast abscess in a woman with normal immune function . The only previous reports of Corynebacterium striatum mastitis have been in cows. J Dairy Sci, 1997 Nov, 80(11), 2815 - 9 Persistence of subclinical intramammary pathogens in goats throughout lactation; Contreras A et al.; The goal of this study was to determine the persistence of caprine intramammary pathogens throughout lactation and to detect the bias in diagnoses when a single milk sample was used . We studied 131 goats throughout 7 mo of lactation . Goats were sampled monthly, and 1834 milk samples were bacteriologically analyzed . One hundred sixty-eight pathogens were isolated: 82.5% were micrococci, 9.5% were Gram-negative bacilli, and 8% were corynebacteria . An intramammary infection (IMI) was considered a true, persistent IMI when the same pathogen was isolated two or more times consecutively from the same half of the udder . One hundred one samples were considered to be truly positive, which produced persistent IMI caused by nine different species (eight Staphylococcus spp . and one Pseudomonas sp.) . Statistical relationships were found between staphylococci and true-positive diagnosis and between corynebacteria and false-positive diagnosis . No relationship involving Gram-negative bacilli was detected . A single milk sample had a positive predictive value (60%), high sensitivity (96.2%), high specificity (96.1%), and highly negative predictive value (99.8%). Res Microbiol, 1997 Jan, 148(1), 45 - 54 Analysis of heterogeneity of Corynebacterium diphtheriae toxin gene, tox, and its regulatory element, dtxR, by direct sequencing; Nakao H et al.; The largest diphtheria outbreak in the developed world since the 1960s is in progress in the Russian Federation . Seventy-two Corynebacterium diphtheriae strains from throughout Russia and the Ukraine, selected for temporal and geographic diversity, and 6 reference and control strains were assayed by DNA direct sequencing, and DNA sequences of their diphtheria toxin gene, tox, and the regulatory dtxR gene, were compared to those of the Park-Williams no . 8 strain (PW8) . Twenty-eight C . diphtheriae strains had entire tox sequences identical to that of the PW8 strain . Among the remaining 40 strains which were toxigenic, 4 point mutations were detected in the tox gene, one within the A and three within the B subunit gene . All four were silent mutations, indicating that diphtheria toxin is highly conserved at the amino acid sequence level; therefore, changes in the efficacy of the current vaccines would be unlikely to occur . Within the open reading frame of the regulatory dtxR gene, 35 point mutations were detected . Only 15 strains had entire dtxR sequences identical to that of the PW8 strain . Nine amino acid substitutions were found in the carboxyl half of dtxR: 22 and 25 strains differed from the PW8 strain in one and two amino acids, respectively . Given that naturally occurring variations of dtxR might be associated with increased diphtheria toxin production, studies to investigate the association of these point mutations and amino acid substitutions with quantified toxin production in the strains causing the current epidemic are under way. J Clin Microbiol, 1997 Dec, 35(12), 3147 - 9 Reversed passive latex agglutination assay for detection of toxigenic Corynebacterium diphtheriae; Toma C et al.; A reversed passive latex agglutination (RPLA) assay for determining the toxigenicity of Corynebacterium diphtheriae is presented . Rabbit antitoxin antiserum was raised by using commercially available diphtheria toxoid . This antiserum reacted with the diphtheria toxin when the culture supernatant was assayed by Western blotting, and it did not cross-react with other extracellular antigens . Affinity-purified antibodies for latex sensitization were obtained by using a Hi Trap N-hydroxysuccinimide-activated column . Demonstration of toxin in five of seven clinical isolates was in accordance with the PCR assay and the Vero cell cytotoxicity test . Culture of the bacteria for 6 h was sufficient for toxin production, and an additional 6 h was needed to observe latex agglutination . Therefore, diphtheria toxin can be detected in 12 h by this method . The lowest concentration of diphtheria toxin detectable by the RPLA assay was about 5 ng/ml . The RPLA assay can provide a convenient and reliable method for laboratories involved in the identification of toxinogenic corynebacteria. J Clin Microbiol, 1997 Dec, 35(12), 3122 - 6 Multicenter evaluation of the updated and extended API (RAPID) Coryne database 2.0; Funke G et al.; In a multicenter study, 407 strains of coryneform bacteria were tested with the updated and extended API (RAPID) Coryne system with database 2.0 (bioMerieux, La-Balme-les-Grottes, France) in order to evaluate the system's capability of identifying these bacteria . The design of the system was exactly the same as for the previous API (RAPID) Coryne strip with database 1.0, i.e., the 20 biochemical reactions covered were identical, but database 2.0 included both more taxa and additional differential tests . Three hundred ninety strains tested belonged to the 49 taxa covered by database 2.0, and 17 strains belonged to taxa not covered . Overall, the system correctly identified 90.5% of the strains belonging to taxa included, with additional tests needed for correct identification for 55.1% of all strains tested . Only 5.6% of all strains were not identified, and 3.8% were misidentified . Identification problems were observed in particular for Corynebacterium coyleae, Propionibacterium acnes, and Aureobacterium spp . The numerical profiles and corresponding identification results for the taxa not covered by the new database 2.0 were also given . In comparison to the results from published previous evaluations of the API (RAPID) Coryne database 1.0, more additional tests had to be performed with version 2.0 in order to completely identify the strains . This was the result of current changes in taxonomy and to provide for organisms described since the appearance of version 1.0 . We conclude that the new API (RAPID) Coryne system 2.0 is a useful tool for identifying the diverse group of coryneform bacteria encountered in the routine clinical laboratory. Infect Immun, 1997 Dec, 65(12), 5364 - 7 Characterization of lipoprotein IRP1 from Corynebacterium diphtheriae, which is regulated by the diphtheria toxin repressor (DtxR) and iron; Schmitt MP et al.; The Corynebacterium diphtheriae irp1 gene is negatively regulated by DtxR and iron . The nucleotide sequence of irp1 revealed that it has homology with genes involved in iron acquisition . Expression of the irp1 gene showed that it encodes a lipoprotein (IRP1) with a predicted size of 38 kDa . Northern blot experiments indicated that transcription from the irp1 promoter is repressed in high-iron medium and suggested that irp1 is part of an iron-regulated operon. Curr Opin Pulm Med, 1996 Jul, 2(4), 335 - 40 Pleural malignancies; Vargas FS et al.; Carcinoma of the lung, metastatic breast carcinoma, and lymphoma are responsible for approximately 75% of all malignant pleural effusions . The presence of malignant cells in the pleural fluid or in the parietal pleura confirms the diagnosis . Recently, several authors have proposed the combination of morphometric procedures and quantitative analysis of nucleolar organizer regions stained by silver nitrate . Videothoracoscopy is recommended for patients suspected of having a malignant pleural effusion in whom the diagnosis is not established after two cytologic studies of the fluid and one needle biopsy . The standard treatment is the intrapleural instillation of a chemical agent to produce a pleurodesis . The recommended sclerosant is talc, a tetracycline derivative, or Corynebacterium parvum where it is available . When a patient is not an ideal candidate for chemical pleurodesis, the options include symptomatic treatment, serial thoracentesis, implantation of a pleuroperitoneal shunt, and pleurectomy. Appl Environ Microbiol, 1997 Nov, 63(11), 4392 - 400 Heterologous expression of the Mycobacterium tuberculosis gene encoding antigen 85A in Corynebacterium glutamicum; Salim K et al.; By using appropriate Corynebacterium glutamicum-Escherichia coli shuttle plasmids, the gene encoding the fibronectin-binding protein 85A (85A) from Mycobacterium tuberculosis was expressed in C . glutamicum, also an actinomycete and nonsporulating gram-positive rod bacterium, which is widely used in industrial amino acid production . The 85A gene was weakly expressed in C . glutamicum under the control of the ptac promoter from E . coli, but it was produced efficiently under the control of the promoter of the cspB gene encoding PS2, one of the two major secreted proteins from C . glutamicum . The 85A protein was produced in various forms, with or without its own signal sequence and with or without the signal sequence and the NH2-terminal (18-amino-acid) mature sequence of PS2 . Western blot analysis with monoclonal antibodies raised against the M . tuberculosis antigen 85 complex showed that recombinant 85A protein was present in the corynebacterial cell wall extract and also released in extracellular culture medium . NH2-terminal microsequencing of recombinant 85A secreted by C . glutamicum showed that signal peptide was effectively cleaved off at the predicted site . The recombinant 85A protein was biologically active in vitro, inducing significant secretion of Th1 T-cell cytokines, particularly interleukin-2 and gamma interferon, in spleen cell cultures from mice vaccinated with live Mycobacterium bovis BCG . Heterologous expression of mycobacterial antigens in C . glutamicum now offers a potent tool for further immunological characterization and large scale preparation of these recombinant proteins. Appl Environ Microbiol, 1997 Nov, 63(11), 4185 - 90 Purification and characterization of staphylococcin BacR1, a broad-spectrum bacteriocin; Crupper SS et al.; The bacteriocin BacR1 was purified from culture supernatant of Staphylococcus aureus UT0007 by sequential ammonium sulfate precipitation, cation-exchange chromatography, and C4 reverse-phase chromatography steps . Mass spectrographic analysis indicated that the purified peptide has a molecular mass of 3,338 Da . It is resistant to environmental conditions, retaining full biological activity after exposure to pH extremes (pHs 3 to 11), heating at 95 degrees C for 15 min, and exposure to strong chaotropic agents . BacR1 was destroyed with a complete loss of biological activity after digestion with trypsin and proteinase K . Amino acid sequence analysis revealed a high concentration of Asx, Gly, and Pro residues and a high proportion of hydrophobic amino acids . The peptide is bactericidal and kills in a dose-dependent manner, but it does not lyse log-phase cells of Corynebacterium renale, the routine indicator organism for bacteriocin assay . A specific receptor for binding was detected on sensitive cells but not on insensitive cells . Competition assays showed that UV-inactivated cells could protect susceptible cells from antibacterial action . A partial inhibitory spectrum revealed that organisms from the following genera are susceptible: Staphylococcus, Streptococcus, Corynebacterium, Haemophilus, Bordetella, Moraxella, Pasteurella, Neisseria, and Bacillus. Prev Vet Med, 1997 Sep, 32(1-2), 77 - 93 Effect of different sampling techniques on odds ratio estimates using hospital-based cases and controls; Doherr MG et al.; Potential biases introduced by the use of hospital admission records have rarely been discussed in the veterinary literature . Veterinary Medical Teaching Hospital (VMTH) patient records kept at the University of California, Davis (UCD) School of Veterinary Medicine provide a unique opportunity to perform in-depth analyses on the effect of different control selection (sampling) techniques on odds ratio (OR) estimates for disease risk factors in a retrospective case-control study . Horses with Corynebacterium pseudotuberculosis abscesses (134) and the (secondary) study base population (source for controls) were identified, and a 'gold standard' OR for each category of the factors admission type, age, breed and sex was derived . Example data were used to calculate sampling ratios (SRs), defined as the ratio between any sample proportion (of an arbitrary risk factor) and the study base proportion for this risk factor . Sampling ratios different from 1.0 introduced biases into the observed OR estimates, when compared with the 'gold standard' OR . Three randomized samples (simple random, stratified random, systematic sampling), one matched (on date of admission) and three different diagnosis samples ('colic', 'cuts and lacerations', 'fractures') were selected from the study base, and the SRs for all categories of the four factors were derived . The matched and two different disease samples ('colic' and 'fractures') had especially wide ranges of observed SRs (and large errors in the OR estimates), whereas simple random and systematic sampling had comparably narrow ranges (less biased OR estimates) . For the three randomized sampling techniques under study, repeated sampling was used to derive SR distributions . The SRs were approximately normally distributed . Analysis of variance and covariance showed that simple random and systematic sampling provided SR distributions with means closest to 1.0 (expected value) and small standard deviations . The OR estimates obtained from records selected by these two sampling techniques therefore were least biased . The findings demonstrate the importance of selecting appropriate sampling techniques in addition to properly defining the study (base) population . Sampling design introduces uncertainty into the OR estimates . The direction of the bias, however, depends on the OR between factor and disease in the source population (the 'gold standard'), and on the direction and magnitude of the SR . When combining the results from single and repeated sampling we conclude that sampling design is most influential on the range of the observed SRs (single samples), on the absolute deviation of the SR from 1.0 (expressed as SR delta Mean) and on the SR standard deviation (SD) (repeated sampling). J Cataract Refract Surg, 1997 Sep, 23(7), 1122 - 5 Chronic pseudophakic endophthalmitis versus saccular endophthalmitis; Abreu JA et al.; A retrospective survey of 1456 cataract operations done at one hospital from 1991 to 1994 found three cases of chronic pseudophakic endophthalmitis . In two of these cases and one referred from another center, microbiological studies from aqueous humor, capsular bag, and vitreous humor were performed using an identical technique . The histopathology in the three cases showed the presence of microorganisms in the capsular wall, although the cultures were only positive in one case (for corynebacterium species). J Cataract Refract Surg, 1997 Sep, 23(7), 1064 - 9 Anterior chamber contamination during cataract surgery with intraocular lens implantation; Mistlberger A et al.; PURPOSE: To measure anterior chamber bacterial and fungal contamination at the beginning and end of cataract surgery with intraocular lens (IOL) implantation in a large series of patients and to determine the influence of preoperative treatment and operative technique on contamination . SETTING: Department of Ophthalmology, County Hospital of Salzburg, Austria . METHODS: This prospective study comprised 700 consecutive patients having planned cataract extraction (511 phacoemulsification, 189 extracapsular cataract extraction {ECCE}) . Thirty-four patients required an anterior vitrectomy; 8 myopic patients did not receive an IOL . A preoperative smear and two intraoperative (at the beginning and end of surgery) anterior chamber aspirates were obtained from each patient . Postoperative smears were obtained at discharge . Three preoperative treatments were evaluated: no lacrimal system irrigation, no topical antibiotic (n = 282); lacrimal system irrigation with balanced saline solution, no topical antibiotic (n = 243); lacrimal system irrigation, antibiotic (neomycin) eyedrops (n = 175) . All patients received topical indomethacin twice a day preoperatively . RESULTS: Preoperative conjunctival smears showed bacterial growth in 76.6% of eyes, with coagulase-negative staphylococci (75%) the most common bacteria . Anterior chamber aspirates were culture positive in 14.1% at the beginning and in 13.7% at the end of surgery, with coagulase-negative staphylococci and corynebacteria the most common . Contamination rates of conjunctival smears taken at discharge were significantly lower (35%) than those taken preoperatively . There was no statistically significantly higher risk of anterior chamber contamination in eyes having ECCE than in those having phacoemulsification . Preoperative treatment did not statistically significantly influence intraoperative aqueous humor contamination rates . There were no cases of acute postoperative endophthalmitis . CONCLUSION: Bacteria entered the anterior chamber during cataract extraction and remained there at the end of surgery in a significant percentage of patients . Surgical technique, preoperative antibiotics, and preoperative lacrimal system irrigation had no statistically significant effect on contamination. Infect Immun, 1997 Nov, 65(11), 4634 - 41 Transcription of the Corynebacterium diphtheriae hmuO gene is regulated by iron and heme; Schmitt MP; The hmuO gene is required for the utilization of heme and hemoglobin as iron sources by Corynebacterium diphtheriae . The product of hmuO has homology to eukaryotic heme oxygenases which are involved in the degradation of heme and the release of iron . To investigate the mechanism of hmuO regulation, a promoterless lacZ gene present on the promoter-probe vector pCM502 was placed under transcriptional control of the hmuO promoter . In C . diphtheriae C7, optimal expression from the hmuO promoter was obtained only in the presence of heme or hemoglobin under low-iron conditions . Expression of hmuO in high-iron medium containing heme was repressed five- to sixfold from that seen under low-iron conditions in the presence of heme . Transcription from the hmuO promoter in the absence of heme or hemoglobin was fully repressed in high-iron medium and was expressed at very low levels in iron-depleted conditions . Expression studies with tile hmuO-lacZ fusion construct in C7hm723, a dtxR mutant of C7, and in a hmuO mutant of C . diphtheriae HC1 provided further evidence that transcription of the hmuO promoter is repressed by DtxR and iron and activated by heme . In Escherichia coli, the hmuO promoter was expressed at very low levels under all conditions examined . Gel mobility shift assays and DNase I footprinting experiments indicated that DtxR binds in a metal-dependent manner to a sequence that overlaps the putative hmuO promoter . Total cellular RNA isolated from C . diphtheriae was used to identify the transcriptional start site for the hmuO gene . Northern blot analysis suggested that the hmuO mRNA was monocistronic and that transcription was heme inducible. Appl Environ Microbiol, 1997 Oct, 63(10), 3783 - 8 Purification and characterization of phenylacetaldehyde reductase from a styrene-assimilating Corynebacterium strain, ST-10; Itoh N et al.; A novel phenylacetaldehyde reductase was purified about 50-fold to homogeneity from Corynebacterium sp . strain ST-10, which can assimilate gaseous styrene as the sole carbon and energy source . The enzyme was inductively synthesized when grown on gaseous styrene and had an important role in styrene metabolism in vivo . The enzyme had a molecular weight of 155,000 and was composed of four identical subunits (molecular weight, 42,000) . The enzyme catalyzed the reduction of not only phenylacetaldehyde but also various aldehydes and ketones; however, it did not catalyze the reverse reaction, the dehydrogenation of 2-phenylethanol . The enzyme required NADH as a cofactor and showed no activity with NADPH; therefore, it was defined as an NADH-dependent phenylacetaldehyde reductase . The enzyme stereospecifically produced (S)-(-)-1-phenylethanol from acetophenone; therefore, it would be useful as a biocatalyst. Eur J Clin Microbiol Infect Dis, 1997 Aug, 16(8), 610 - 4 Comparison of traditional and molecular methods for typing nontoxigenic strains of Corynebacterium diphtheriae; Riegel P et al.; Thirty-eight nontoxigenic strains of Corynebacterium diphtheriae isolated between 1987 and 1992 from clinical specimens of French patients were typed by biotyping, antibiograms, bacteriophage typing, ribotyping, and restriction analysis by pulsed-field gel electrophoresis (PFGE) . Excellent correlation occurred between the genotypes defined by PFGE SfiI profiles or by ribotype BstEII profiles . Genotyping revealed seven genotype patterns among the 26 biotype mitis isolates, five among the nine biotype gravis isolates, and three among the three biotype belfanti isolates . Phage typing was nonreactive for nine of the 38 isolates . A combination of all the typing methods led to the identification of 19 different types of Corynebacterium diphtheriae. Microbiol Immunol, 1997, 41(9), 649 - 55 Phylogenetic positions and assignment of swine and ovine corynebacterial isolates based on the 16S rDNA sequence; Takahashi T et al.; The nucleotide sequences of the 16S ribosomal RNA gene (rDNA) of swine and ovine corynebacterial strains were determined . The sequences of the strains that identified as Corynebacterium pseudotuberculosis by their biochemical characteristics were homologous with each other . The phylogenetic position of C . pseudotuberculosis strains was closet to C . ulcerans and next closet to C . diphtheriae . The nucleotide sequence of another swine isolate, SC8, was similar to that of a recently proposed species, C . seminale, and a non-validated species, "C . glucuronolyticum," with about 0.01 to 0.02 evolutionary distances . Analysis of the predicted secondary structure of the 16S rRNA molecule agreed with the close phylogenetic relationships between C . pseudotuberculosis and C . ulcerans and between C . seminale and strain SC8. Int J Syst Bacteriol, 1997 Oct, 47(4), 1107 - 11 Corynebacterium durum sp . nov., from human clinical specimens; Riegel P et al.; A new Corynebacterium species, Corynebacterium durum, was isolated from respiratory tract specimens of five human patients . The strains of this species exhibited similar morphologic and biochemical features that differentiated them from all recognized species . Notably, all of these strains developed irregular and strongly adherent colonies under aerobic conditions and produced acid from mannitol and galactose . The cells are long pleomorphic rods with some filaments . This species has characteristics of the genus Corynebacterium, such as 55 mol% guanine plus cytosine in the DNA and the presence of corynomycolic acids, meso-diaminopimelic acid, arabinose, and galactose in the cell wall . These isolates formed a homogeneous group in which the DNA-DNA similarity values (as determined by an S1 nuclease procedure) compared with reference strain IBS G15036T (T = type strain) ranged from 71 to 100% . The analysis of the nearly complete 16S rRNA gene sequence of IBS G15036T indicated that this new species represents a distinct taxon within the genus Corynebacterium . This new species can be identified on the basis of its colony morphology, fermentation of sugars, and enzymatic activities . Strain IBS G15036 (= CCUG 37331) is the type strain of C . durum. Int J Syst Bacteriol, 1997 Oct, 47(4), 1092 - 6 Corynebacterium singulare sp . nov., a new species for urease-positive strains related to Corynebacterium minutissimum; Riegel P et al.; We studied two coryneform strains from clinical specimens . These strains had type IV and corynemycolic acids in their cell walls and also had phenotypic characteristics, such as urease activity and fermentation of glucose and sucrose but not trehalose, which did not permit assignment to any previously recognized taxon . According to DNA-DNA hybridization data, these two strains are members of the same species (level of DNA similarity, 86%) . Phylogenetic analysis based on comparisons of almost complete small-subunit ribosomal DNA sequences revealed that these strains are closely related to Corynebacterium minutissimum, but DNA relatedness experiments clearly showed that they constitute a distinct new species with a level of DNA relatedness to the C . minutissimum type strain of less than 40% . This new species can be differentiated from C . minutissimum strains by its enzymatic activities and carbon source utilization, and the name Corynebacterium singulare is proposed for it . The type strain is strain IBS B52218 (= CCUG 37330), which was isolated from a semen specimen. Int J Syst Bacteriol, 1997 Oct, 47(4), 1082 - 5 Corynebacterium mastitidis sp . nov., isolated from milk of sheep with subclinical mastitis; Fernandez-Garayzabal JF et al.; Fourteen strains of a hitherto unknown catalase-positive, aerobic, gram-positive coryneformlike organism were isolated from the milk of sheep with subclinical mastitis from different regions of Spain . The strains phenotypically closely resembled one another and biochemically were similar to Corynebacterium urealyticum and Corynebacterium afermentans subsp . lipophilum . The results of chemotaxonomic investigations were consistent with membership in the genus Corynebacterium, and comparative 16S rRNA gene sequencing studies showed that the unknown bacterium from sheep was indeed a member of the genus Corynebacterium . Within the genus Corynebacterium the new bacterium formed a distinct subline that exhibited > 4% sequence divergence with other species . Based on both phenotypic and phylogenetic findings, a new species, Corynebacterium mastitidis, is proposed for the organisms from mastitic sheep . The type strain of C . mastitidis is CECT 4843 (= S-8). Int J Syst Bacteriol, 1997 Oct, 47(4), 952 - 7 Corynebacterium mucifaciens sp . nov., an unusual species from human clinical material; Funke G et al.; Eight strains of a previously undescribed coryneform bacterium had been isolated from human clinical material over a 5-year period . Colonies of the unknown coryneform bacterium had an unusual appearance as they were slightly yellowish and very mucoid . Biochemical and chemotaxonomic characterization revealed that the unknown coryneform bacterium belonged to the genus Corynebacterium . It could be readily differentiated from all previously described Corynebacterium species . Electron microscopy demonstrated the production of an extracellular substance causing connecting filaments between cells as a morphological correlate to the mucoid colonies . Comparative 16S rRNA gene sequence analysis revealed that the unknown coryneform bacterium represented a new subline within the genus Corynebacterium, for which the name Corynebacterium mucifaciens sp . nov . is proposed . The type strain is CCUG 36878 (= DSM 44265 = CIP 105129). Biochem J, 1997 Sep 15, 326 ( Pt 3), 773 - 83 alpha-1,4-D-glucan phosphorylase of gram-positive Corynebacterium callunae: isolation, biochemical properties and molecular shape of the enzyme from solution X-ray scattering; Weinhausel A et al.; The alpha-1,4-D-glucan phosphorylase from gram-positive Corynebacterium callunae has been isolated and characterized . The enzyme is inducible approx . 2-fold by maltose, but remarkably not repressed by D-glucose . The phosphorylase is a homodimer with a stoichiometric content of the cofactor pyridoxal 5'-phosphate per 88-kDa protein subunit . The specificity constants (kcat/Km, glucan) in the directions of glucan synthesis and degradation are used for the classification of the enzyme as the first bacterial starch phosphorylase . A preference for large over small substrates is determined by variations in the apparent binding constants rather than catalytic-centre activities . The contribution of substrate chain length to binding energy is explained assuming two glucan binding sites in C . callunae phosphorylase: an oligosaccharide binding site composed of five subsites and a high-affinity polysaccharide site separated from the active site . A structural model of the molecular shape of the phosphorylase was obtained from small-angle solution X-ray scattering measurements . A flat, slightly elongated, ellipsoidal model with the three axes related to each other as 1:(0.87-0.95):0.43 showed scattering equivalence with the enzyme molecule . The model of C . callunae phosphorylase differs from the structurally well-characterized rabbit-muscle phosphorylase in size and axial dimensions. Infect Immun, 1997 Oct, 65(10), 4273 - 80 Identification and characterization of three new promoter/operators from Corynebacterium diphtheriae that are regulated by the diphtheria toxin repressor (DtxR) and iron; Lee JH et al.; DtxR is a dimeric, sequence-specific, DNA-binding protein that functions as an iron-dependent, negative global regulator in Corynebacterium diphtheriae . Under high-iron conditions, DtxR represses the synthesis of diphtheria toxin, corynebacterial siderophore, and other components of the high-affinity iron uptake system . Three DtxR-regulated promoter/operators designated tox, IRP1, and IRP2 were reported previously . In this study, we identified and characterized three additional DtxR-regulated promoter/operators from C . diphtheriae designated IRP3, IRP4, and IRP5 . When beta-galactosidase was expressed from these three new promoter/ operators in Escherichia coli containing dtxR+ on pDSK29, enzyme levels were 5- to 30-fold lower during high-iron growth than during low-iron growth . In gel shift assays, the mobility of DNA fragments containing each promoter/operator decreased in the presence of purified DtxR and Co2+ . In footprinting assays, DtxR protected 36-, 35-, and 30-bp regions of IRP3, IRP4, and IRP5, respectively, from cleavage by DNase I . In the 19-bp core of each promoter/operator, 12 or 13 bp matched the consensus for the DtxR-binding site . The putative polypeptides encoded by the open reading frames (ORFs) downstream from IRP3 and IRP4 were homologous, respectively, to several bacterial transcriptional regulators and to the deduced polypeptide encoded by an ORF located between the E . coli genes for primosomal replication protein N and adenine phosphoribosyltransferase . The putative polypeptide encoded by the ORF downstream from IRP5 was not homologous to any sequence in the protein database at the National Center for Biotechnology Information . When the ORFs downstream from IRP3 and IRP4 were expressed under the control of the phage T7 promoter in E . coli, polypeptide products of the predicted sizes were detected in small amounts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Rev Alerg Mex, 1993 Sep-Oct, 40(5), 110 - 3 {Clinical and laboratory features of 100 patients with perennial allergic rhinitis}; Segura Mendez NH et al.; 100 patient records were studied with a clinical diagnosis of perennial allergic rhinitis; they were 66 women and 34 men, with a range of 28.6 years old . The cutaneous tests were positive to pollens in 78% of the cases, fungus 39%, inhalables 39%, Dermatophagoides 19% and bacterial 7% . In the nasal mucous culture the following germs were isolated: S epidermidis 49%, S aureus 25%, Neisseria sp 15%, Corynebacterium 2%, P mirabilis 1% and E coli 1% . The nasal cytology was positive in 25% of the cases for the presence of eosinophils, and was negative in 75% . In only 27% of the patients eosinophil was found in the peripheric blood . The results are commented and the utility of the cultivation of nasal mucous and of the cytology of nasal mucous in patients with perennial allergic rhinitis is discussed. Int J Biochem Cell Biol, 1997 Jun, 29(6), 895 - 900 Cloning and characterization of the genes of the CeqI restriction-modification system; Izsvak Z et al.; Two genes from Corynebacterium equii, a Gram-positive bacterium producing the CeqI restriction-modification enzymes were cloned and sequenced . In vivo restriction experiments, DNA and amino acid sequence data suggest that the two genes code for the endonuclease and the methyltransferase enzymes . However, when the two genes are expressed in E . coli, practically no enzyme activity can be detected in the supernatants of sonicated cells . Based on the DNA sequence data CeqI restriction endonuclease (an EcoRV izoschizomer) consists of 270 amino acid residues with a predicted molecular mass of 31.6 kDa, in good agreement with the previously measured 32 +/- 2 kDa . The methyltransferase is 517 residues long (approx . 60 kDa) . The two genes are in opposite orientation and overlap by 37 base pairs on the chromosome . The deduced amino acid sequence of the putative endonuclease gene revealed long stretches of hydrophobic amino acids, that may form the structural basis of the unusual aggregation properties of the restriction endonuclease . The amino acid sequence of the methylase shows homologies with other type II methyltransferases. FEMS Microbiol Lett, 1997 Sep 1, 154(1), 81 - 8 Isolation of the Corynebacterium glutamicum glnA gene encoding glutamine synthetase I; Jakoby M et al.; The Corynebacterium glutamicum glutamine synthetase I (GSI) structural gene glnA was cloned by a PCR approach using oligonucleotide primers derived from conserved amino acid sequences of the GSI proteins from various bacteria . Disruption or deletion of this gene in C . glutamicum led to a glutamine auxotrophic phenotype and complete loss of glutamine synthetase activity, indicating the key role of this enzyme in nitrogen metabolism . Additionally, indications for a second glutamine synthetase, GSII, were found. Arch Microbiol, 1997 Oct, 168(4), 262 - 9 Regulation of acetate metabolism in Corynebacterium glutamicum: transcriptional control of the isocitrate lyase and malate synthase genes; Wendisch VF et al.; In the amino-acid-producing microorganism Corynebacterium glutamicum, the specific activities of the acetate-activating enzymes acetate kinase and phosphotransacetylase and those of the glyoxylate cycle enzymes isocitrate lyase and malate synthase were found to be high when the cells were grown on acetate (0.8, 2.9, 2.1, and 1.8 U/mg protein, respectively) . When the cells were grown on glucose or on other carbon sources such as lactate, succinate, or glutamate, the specific activities were two- to fourfold (acetate kinase and phosphotransacetylase) and 45- to 100-fold (isocitrate lyase and malate synthase) lower, indicating that the synthesis of the four enzymes is regulated by acetate in the growth medium . A comparative Northern (RNA) analysis of the C . glutamicum isocitrate lyase and malate synthase genes (aceA and aceB) and transcriptional cat fusion experiments revealed that aceA and aceB are transcribed as 1.6- and 2.7-kb monocistronic messages, respectively, and that the regulation of isocitrate lyase and malate synthase synthesis is exerted at the level of transcription from the respective promoters . Surprisingly, C . glutamicum mutants defective in either acetate kinase or phosphotransacetylase showed low specific activities of the other three enzymes (phosphotransacetylase, isocitrate lyase, and malate synthase or acetate kinase, isocitrate lyase, and malate synthase, respectively) irrespective of the presence or absence of acetate in the medium . This result and a correlation of a high intracellular acetyl coenzyme A concentration with high specific activities of isocitrate lyase, malate synthase, acetate kinase, and phosphotransacetylase suggest that acetyl coenzyme A or a derivative thereof may be a physiological trigger for the genetic regulation of enzymes involved in acetate metabolism of C . glutamicum. Rev Inst Med Trop Sao Paulo, 1996 Sep-Oct, 38(5), 359 - 63 Effects of non-specific immunopotentiators in experimental Schistosoma mansoni infection . II . Corynebacterium parvum; Teixeira KM et al.; The effects of Corynebacterium parvum on host protection, tissue reaction and "in vivo" chemotaxis in Schistosoma mansoni infected mice were studied . The C . parvum was given intraperitoneally using a dose of 0.7 mg, twice a week (for 4 weeks), thirty days before (prophylactic treatment) or after infection (curative treatment) . The host protection was evaluated through the recovery of adult worms by liver perfusion and was lower in the prophylactic group as compared to the control group (p = 0.018), resulting in 44% protection . The "in vivo" leukocyte response in both prophylactic and curative groups was higher as compared to the infected/non treated group (p = 0.009 and p = 0.003, respectively) . Tissue reactions were described in the experimental and control groups, but there were not remarkable differences among them . The possible biological implications and relevance of the findings for the defensive response of the host and control of schistosomiasis are discussed. J Immunol, 1997 Sep 1, 159(5), 2462 - 7 Chitin particle-induced cell-mediated immunity is inhibited by soluble mannan: mannose receptor-mediated phagocytosis initiates IL-12 production; Shibata Y et al.; Previous studies showed that mouse spleen cells produced IL-12, TNF-alpha, and IFN-gamma when stimulated with phagocytosable-size chitin particles (N-acetyl-D-glucosamine polymers) . To dissect the mechanisms of the cytokine production in this study, spleen cells from BALB/c mice were cultured with 1 to 10 microm chitin particles, heat-killed Corynebacterium parvum vaccine, zymosan, and mannan (a mannose polymer)-coated latex beads (1 microm) at 1, 10, or 100 microg/ml . We found that these particles induced IL-12, TNF-alpha, and IFN-gamma . However, these cytokines were not produced when spleen cells were cultured with soluble chitin, mannan, or laminarin (a polymer of beta-glucan), 1 to 10 microm beta-glucan particles, laminarin-coated latex beads, 1 microm latex beads, 50 to 100 microm chitin particles, or 50 to 100 microm mannan-coated beads . Soluble mannan, but not soluble laminarin, inhibited cytokine production following stimulation with 1 to 10 microm chitin particles, zymosan, or heat-killed C . parvum . In addition, cytochalasin D also inhibited cytokine production . The treatments with soluble mannan or with cytochalasin D, in sharp contrast, did not inhibit LPS-induced IL-12/IFN-gamma production or exogenous IL-12-induced IFN-gamma production . Finally, spleen cells from C3H/HeJ mice also showed comparable levels of IL-12/TNF-alpha/IFN-gamma production when induced by 1 to 10 microm chitin particles . Taken together, our results indicate that mannose receptor-mediated phagocytosis, but not the receptor-mediated pinocytosis, is highly associated with the production of IFN-gamma-inducing extracellular signaling factors such as IL-12 and TNF-alpha . The novel mechanism of phagocytosis-dependent IL-12 production appears to be distinct from that of LPS-induced cytokine production. Am J Physiol, 1997 Aug, 273(2 Pt 1), G530 - 6 Inhibition of nitric oxide synthesis improves detoxication in inflammatory liver dysfunction in vivo; Veihelmann A et al.; Inflammatory stimulation of the liver induces nitric oxide (NO) biosynthesis and suppression of detoxication . In this study the effect of NO biosynthesis on cytochrome P-450 (CYP) enzyme activity was investigated by comparing in vivo and in vitro assays . To establish liver inflammation, CD rats were injected with Corynebacterium parvum (C . parvum) suspension . After 5 days NO biosynthesis was highly induced as indicated by increased NO2- plus NO3- serum concentrations . At the same time the aminopyrine breath test (ABT), measuring CYP activity in vivo, was reduced to 42% and the in vitro assay of aminopyrine turnover was suppressed to 12% of NaCl- injected controls . When C . parvum-injected animals were treated with the NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA), CYP activities significantly improved with an ABT of 76% and an in vitro aminopyrine turnover of 47% of controls . Neither C . parvum injections nor L-NMMA treatment resulted in a significant change of CYP protein concentrations . These data indicate that suppression of xenobiotic metabolism can be attenuated by inhibition of NO biosynthesis during an ongoing process of inflammation. J Dairy Sci, 1997 Aug, 80(8), 1592 - 9 Estimation of interdependence among quarters of the bovine udder with subclinical mastitis and implications for analysis; Barkema HW et al.; Interdependency among udder quarters with subclinical mastitis was evaluated on 150 farms using a total of 35,828 udder quarters . The occurrence of high somatic cell count (SCC) (> 250,000 cells/ml) in 0, 3, and 4 quarters occurred at a higher rate than would be expected based on independence of the quarters . For all bacterial species, intramammary infection in 0, 2, 3, or 4 quarters of the same cow occurred at a higher rate than would be expected based on independence of the quarters . Intramammary infection and high SCC were found less often in front quarters than in rear quarters . High SCC and intramammary infection occurred more often in right front quarters than in left front quarters . High SCC in diagonal quarters occurred at a lower rate than expected . Corynebacterium bovis, Streptococcus agalactiae, and Staphylococcus aureus had the highest intraclass correlation within herd . Streptococcus uberis had a very low intraclass correlation within herd . The intraclass correlation within cow for the natural logarithm of SCC was 0.47 . Corynebacterium bovis and Strep . agalactiae had the highest intraclass correlation within cow, and Streptococcus dysgalactiae had the lowest . Analytical methods were proposed to manage the problem of interdependence and its effect on the design or evaluation of field studies on subclinical mastitis. Neuroradiology, 1997 Aug, 39(8), 581 - 2 Dense Rhodococcus cerebral abscesses in an HIV-positive patient; Perrymore WD et al.; Rhodococcus equi, formerly known as Corynebacterium equi, an aerobic, gram-positive, pleomorphic coccobacillus, is a well-known pathogen for domestic livestock . We present a biopsy- and culture-proven case of Rhodococcus equi brain abscesses in a patient seropositive for HIV, having an appearance not described previously. Arch Microbiol, 1997 Aug, 168(2), 143 - 51 Isolation of the putP gene of Corynebacterium glutamicum and characterization of a low-affinity uptake system for compatible solutes; Peter H et al.; Corynebacterium glutamicum accumulates the compatible solutes proline, glycine betaine, and ectoine under conditions of high osmolality . Uptake of proline is mediated by both a high-affinity and a low-affinity secondary transport system . The low-affinity uptake system also accepts glycine betaine and ectoine as substrates . In the present study, the gene encoding the high-affinity proline uptake system PutP was isolated by heterologous complementation of Escherichia coli mutant strain WG389, which lacks the transport systems BetT, PutP, ProP, and ProU and is unable to synthesize proline and glycine betaine . This gene (putP) encodes a protein of 524 amino acids that shares identity with the proline transport systems PutP of E . coli, Staphylococcus aureus, Salmonella typhimurium, Haemophilus influenzae, and Klebsiella pneumoniae . Functional studies of PutP synthesized in E . coli mutant strain MKH13, which also lacks the transport systems for compatible solutes and is unable to synthesize glycine betaine, revealed that this carrier system is not regulated by the external osmolality on the level of activity . Km values of 7.6 mM for proline and 1.3 mM for sodium as cotransported ion were determined . Deletion of the putP gene allowed the functional characterization of another proline uptake system with low affinity. Infect Immun, 1997 Aug, 65(8), 3048 - 56 Attenuation and vaccine potential of aroQ mutants of Corynebacterium pseudotuberculosis; Simmons CP et al.; Corynebacterium pseudotuberculosis, a gram-positive intracellular bacterial pathogen, is the etiological agent of the disease caseous lymphadenitis (CLA) in both sheep and goats . Attenuated mutants of C . pseudotuberculosis have the potential to act as novel live veterinary vaccine vectors . We have cloned and sequenced the aroB and aroQ genes from C . pseudotuberculosis C231 . By allelic exchange, aroQ mutants of both C231, designated CS100, and a pld mutant strain TB521, designated CS200, were constructed . Infection of BALB/c mice indicated that introduction of the aroQ mutation into C231 and TB521 attenuated both strains . In sublethally infected BALB/c mice, both CS100 and CS200 were cleared from spleens and livers by day 8 postinfection . The in vivo persistence of these strains was increased when the intact aroQ gene was supplied on a plasmid in trans . Mice infected with TB521 harbored bacteria in organs at least till day 8 postinfection without ill effect . When used as a vaccine, only the maximum tolerated dose of CS100 had the capacity to protect mice from homologous challenge . Vaccination with TB521 also elicited protective immunity, and this was associated with gamma interferon (IFN-gamma) production from splenocytes stimulated 7 days postvaccination . The role of IFN-gamma in controlling primary infections with C . pseudotuberculosis was examined in mice deficient for the IFN-gamma receptor (IFN-gammaR(-/-) mice) . IFN-gammaR(-/-) mice cleared an infection with CS100 but were significantly more susceptible than control littermates to infection with C231 or TB521 . These studies support an important role for IFN-gamma in control of primary C . pseudotuberculosis infections and indicate that aroQ mutants remain attenuated even in immunocompromised animals . This is the first report of an aroQ mutant of a bacterial pathogen, and the results may have implications for the construction of aromatic mutants of Mycobacterium tuberculosis for use as vaccines. J Clin Microbiol, 1997 Aug, 35(8), 2003 - 6 Microbiologic and clinical value of primary broth cultures of wound specimens collected with swabs; Silletti RP et al.; In order to assess the microbiologic and clinical value of primary broth culture of wound specimens collected with swabs and submitted to the laboratory in transport medium, we compared the results of primary agar culture with the results of a corresponding primary broth culture for 344 aerobic specimens and 176 anaerobic specimens . While 8.7% (45 of 520) of the specimens yielded organisms from the primary broth culture that were not recovered from the corresponding primary agar culture, only 5.0% (26 of 520) of the specimens yielded organisms from the primary broth culture other than Staphylococcus epidermidis, viridans group streptococci, and Corynebacterium spp . Moreover, the primary broth culture of only 0.6% (3 of 520) of the specimens yielded organisms not recovered from the primary agar culture that caused a change in the therapy of the patient . Our conclusion is that primary broth cultures are unnecessary for the processing of wound specimens properly collected with swabs. J Clin Microbiol, 1997 Aug, 35(8), 1978 - 83 Corynebacterium imitans sp . nov . isolated from patients with suspected diphtheria; Funke G et al.; A 5-month-old boy of a Romanian family traveling via Ukraine to Poland developed a respiratory disease that resembled and that was initially diagnosed as pharyngeal diphtheria . The child recovered after treatment with antidiphtheria antitoxin . A coryneform bacterium had been isolated from a nasopharyngeal specimen from the child and was initially identified as an atypical Corynebacterium diphtheriae strain . Seven adults who had contact with either the child or an adult contact person also developed symptoms of pharyngeal diphtheria, were also treated with antitoxin, and recovered uneventfully . Coryneform bacteria similar to that originating from the index patient were also isolated from the throat swabs of three adults . Detailed biochemical and chemotaxonomic investigations revealed that the coryneform bacteria belonged to the genus Corynebacterium and could be differentiated from all other defined species of this genus . Ribotyping and pulsed-field gel electrophoresis demonstrated that all four patients' isolates were of clonal origin . The diphtheria toxin gene and its product were not detected either by PCR assays or by the Elek test, making a possible disease association of the Corynebacterium more unlikely . Comparative 16S rRNA gene sequence analysis revealed that the coryneform bacterium represented a new subline within the genus Corynebacterium, for which the name Corynebacterium imitans sp . nov . is proposed . The type strain is NCTC 13015 (DSM 44264; CCUG 36877). Proc Natl Acad Sci U S A, 1997 Jul 22, 94(15), 8093 - 8 Characterization of tumor necrosis factor-deficient mice; Marino MW et al.; Although tumor necrosis factor (TNF) initially came to prominence because of its anti-tumor activity, most attention is now focused on its proinflammatory actions . TNF appears to play a critical role in both early and late events involved in inflammation, from localizing the noxious agent and amplifying the cellular and mediator responses at the local site and systemically, to editing (e.g., apoptosis) injured cells or effete immune cells and repairing inflammatory damage . We have generated mice deficient in TNF (TNF-/- mice) and have begun to examine the multiple functions attributed to TNF . TNF-/- mice develop normally and have no gross structural or morphological abnormalities . As predicted, they are highly susceptible to challenge with an infectious agent (Candida albicans), are resistant to the lethality of minute doses of lipopolysaccharide (LPS) following D-galactosamine treatment, have a deficiency in granuloma development, and do not form germinal centers after immunization . Phagocytic activity of macrophages appears relatively normal, as do T cell functions, as measured by proliferation, cytokine release, and cytotoxicity . B cell response to thymus-independent antigens is normal, but the Ig response to thymus-dependent antigen is reduced . Surprisingly, cytokine production induced by LPS appears essentially intact, with the exception of reduced colony-stimulating factor activity . Other unexpected findings coming from our initial analysis are as follows . (i) TNF has low toxicity in TNF-/- mice . (ii) TNF-/- mice show an anomalous late response to heat-killed Corynebacterium parvum . In contrast to the prompt response (granuloma formation, hepatosplenomegaly) and subsequent resolution phase in C . parvum-injected TNF+/+ mice, similarly treated TNF-/- mice show little or no initial response, but then develop a vigorous, disorganized inflammatory response leading to death . These results suggest that TNF has an essential homeostatic role in limiting the extent and duration of an inflammatory process-i.e., an anti-inflammatory function . (iii) In contrast to the expectation that TNF+/+ mice and TNF+/- mice would have identical phenotypes, TNF+/- mice showed increased susceptibility to high-dose LPS lethality, increased susceptibility to Candida challenge, and delayed resolution of the C . parvum-induced inflammatory process, indicating a strong gene dose requirement for different actions of TNF. Eur J Biochem, 1997 Jul 15, 247(2), 572 - 80 Efflux of compatible solutes in Corynebacterium glutamicum mediated by osmoregulated channel activity; Ruffert S et al.; Bacteria respond to hypoosmotic stress by releasing low-molecular-mass solutes in order to maintain constant turgor pressure . We have studied the function of osmoregulated channel(s) in Corynebacterium glutamicum, which are responsible for efflux of various solutes upon sudden decrease in osmotic pressure . The channels preferentially mediated efflux of compatible solutes such as glycine betaine and proline . The release of molecules of similar size, e.g . glutamate or lysine, was restricted, ATP was completely retained even after severe osmotic shock . The cells maintained high cytoplasmic K+ and Na+ concentrations under hypoosmotic shock . Several results suggest that the solute efflux is mediated by a channel and not by a carrier, e.g . by reversal of the glycine betaine uptake systems of C . glutamicum: the release of glycine betaine and proline was extremely fast reaching an efflux rate of 6000 micromol x min(-1) x g dm(-1) or higher; the efflux was not significantly influenced by addition of external transport substrate, e.g . glycine betaine; in spite of an extremely high chemical gradient, no significant efflux under isoosmolar conditions was observed; efflux of solutes was unchanged after full uncoupling of membrane energetics by carbonylcyanide m-chlorophenylhydrazone (CCCP) . These results indicate the presence of an osmoregulated channel in C . glutamicum similar to the mechanosensitive channel(s) of Escherichia coli . The activity of the channel did not depend on the growth conditions, but we observed a tight regulation on the level of activity, i.e . the mechanosensitive channel behaved as a perfect osmometer . By monitoring release of glycine betaine under slow and continuous decrease of the external osmolality, we observed continous efflux whithout a stepwise release of solutes . This resulted in a significant steady-state decrease of the membrane potential. Parasitology, 1997 Jul, 115 ( Pt 1), 21 - 8 Induction of protective immunity and modulation of granulomatous hypersensitivity in mice using PIII, an anionic fraction of Schistosoma mansoni adult worm; Hirsch C et al.; This study was performed in order to define Schistosoma mansoni antigens that are able to function as modulator agents in the granulomatous hypersensitivity to parasite eggs in BALB/c and C57BL/6 mice . A fraction of S . mansoni, designated PIII, derived from adult worm antigen preparation (SWAP) was obtained using anion-exchange chromatography on an FPLC system . Immunization of mice with PIII in the presence of Corynebacterium parvum and Al(OH)3 as adjuvant induced an immune response in this animals as determined by ELISA and spleen cell proliferation assays against S . mansoni antigens SEA, SWAP and PIII . In addition, PIII caused a significant degree of protection against a challenge infection in immunized mice as observed by the decrease on worm burden recovered from the portal system . We also showed that PIII profoundly inhibited the vigorous anamnestic granulomatous response to eggs in the liver and lungs . This suppression correlated with a significant decrease in granuloma size . From these results we conclude that the PIII preparation contains antigens that can mediate protective anti-parasite immunity and downregulate granulomatous hypersensitivity to S . mansoni eggs. Eur J Epidemiol, 1997 Jul, 13(5), 535 - 9 Serological survey on the immunity to diphtheria of the northern Greek population; Souliou E et al.; The recent outbreak of diphtheria in the Newly Independent States (NIS) of the former USSR and the immigration from these high risk areas to Greece prompted us to determine the diphtheria antitoxin levels by enzyme-linked immunosorbent assays (ELISA) in 509 healthy individuals (307 males and 202 females) from northern Greece . The population under study was divided in ten age groups from 1 day to > 60 years old . Diphtheria antitoxin levels of > or = 0.1 IU/ml were considered as protective ones . 44.6% of the examined people were found susceptible . The children up to their twenties seem to be immune to diphtheria in a high proportion (86-88.4%) . The diphtheria antitoxin levels declined sharply above this age (17.6% in the age group 21-30 years old) . The level of protection in adults appeared to be higher in the oldest group (49%) . According to these results, the adults are not properly protected . Booster doses of vaccine for them are recommended to improve the resistance of the northern Greek population from possible infection by toxigenic stains of Corynebacterium diphtheriae, imported or endogenous. Eur J Immunol, 1997 Jul, 27(7), 1657 - 62 Lack of correlation between rejection of tumor cells co-expressing interleukin-2 and B7.1 and vaccine efficiency; Cayeux S et al.; Genetically modifying tumor cells to express a variety of cytokines such as interleukin-2 (IL-2) or the co-stimulatory molecule B7.1 leads to increased immunogenicity and reduced tumorigenicity of tumors in several models with T cells involved in the process . We have previously reported decreased tumorigenicity of the murine plasmacytoma J558L {major histocompatibility complex (MHC) class I+ and class II-} expressing IL-2 or B7.1 . When systemic immunity was analyzed, immunization with either J558-IL2 or J558-B7.1 cells generated moderate protection against unmodified J558L tumor cells, comparable to immunization with a tumor cells/adjuvant Corynebacterium parvum mixture . In this study, we asked whether the co-expression of IL-2 and B7.1 in tumor cells would augment vaccine potency, cytotoxic T lymphocyte (CTL) activity and protective immunity . Rejection of single IL-2 or B7.1 or co-transfected IL-2/B7.1 cells occurred in most syngeneic animals but not in T cell-deficient nude mice, thus confirming that T cells were required for tumor rejection . We knew from previous experiments that CD8+ T cells were responsible for rejection . Surprisingly, immunization with J558-IL2/B7.1 cells followed by challenge with parental J558L caused a reduction in systemic protection as compared to J558-B7.1 or J558-IL2 alone . We examined the mechanism underlying this unexpected result: 6 days after injection of J558-IL2/B7.1 cells, tumor were nearly completely destroyed and were almost devoid of CD8+ cells, while CD8+ cells were increased in both IL-2- and B7.1-transfected tumors . In addition, immunization with J558-IL2/B7.1 tumors had an adverse effect on the generation of CTL . Mice immunized with J558-B7.1 and to a lesser extent J558-IL2 cells mounted a CTL response against J558L cells while, in contrast, no CTL activity could be detected in mice immunized with J558-IL2/B7.1, thus showing a correlation between the absence of CTL activity and the lack of in vivo protection . We demonstrate that "hyperstimulation" of the immune response by genetically modified cancer vaccines can have adverse effects on tumor immunity, even though the mechanism is not yet completely understood. Lab Anim, 1997 Jul, 31(3), 206 - 11 Outbreaks of hyperkeratotic dermatitis of athymic nude mice in northern Italy; Scanziani E et al.; Hyperkeratotic dermatitis of athymic nude mice is an infectious disease caused by a coryneform bacterium . During the spring of 1995, outbreaks of hyperkeratotic dermatitis were observed in several nude mice facilities in northern Italy . In this report we describe the clinical, histopathological and microbiological features of the disease in two different animal facilities . Affected animals showed a typical 'scaly' appearance with small white flakes of material adherent to the skin . In one of the outbreaks (facility 2) the lesions were less severe and involved only limited areas of the body . The infection spread very quickly and the morbidity reached more than 80% in a few days, while the mortality was about 1% . The lesions resolved spontaneously within 7-10 days . Histological examination of affected skin revealed orthokeratotic hyperkeratosis, acanthosis and dermal inflammatory infiltration which were more severe in mice from facility 1 . In Gram-stained sections groups of rods consistent with coryneform bacteria were detectable in the keratin layers covering the epidermal surface . A coryneform bacterium, biochemically typed as Corynebacterium bovis, was isolated from 11 out of 11 mice from facility 1 and from 8 out of 11 mice from facility 2. Appl Environ Microbiol, 1997 Jul, 63(7), 2631 - 6 Phylogeny and classification of bacteria in the genera Clavibacter and Rathayibacter on the basis of 16s rRNA gene sequence analyses; Lee IM et al.; A phylogenetic analysis by parsimony of 16S rRNA gene sequences (16S rDNA) revealed that species and subspecies of Clavibacter and Rathayibacter form a discrete monophyletic clade, paraphyletic to Corynebacterium species . Within the Clavibacter-Rathayibacter clade, four major phylogenetic groups (subclades) with a total of 10 distinct taxa were recognized: (I) species C . michiganensis; (II) species C . xyli; (III) species R . iranicus and R . tritici; and (IV) species R . rathayi . The first three groups form a monophyletic cluster, paraphyletic to R . rathayi . On the basis of the phylogeny inferred, reclassification of members of Clavibacter-Rathayibacter group is proposed . A system for classification of taxa in Clavibacter and Rathayibacter was developed based on restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified 16S rDNA sequences . The groups delineated on the basis of RFLP patterns of 16S rDNA coincided well with the subclades delineated on the basis of phylogeny . In contrast to previous classification systems, which are based primarily on phenotypic properties and are laborious, the RFLP analyses allow for rapid differentiation among species and subspecies in the two genera. Plast Reconstr Surg, 1997 Jul, 100(1), 182 - 96 Microbial growth inside saline-filled breast implants; Young VL et al.; In vitro and in vivo experiments were conducted to determine whether intraluminal saline in breast implants can support the growth of common wound-infecting microorganisms over a prolonged period of time . The bacteria tested were Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Corynebacterium jeikeium, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa . Three fungal species also were tested: Aspergillus fumigatus, Paecilomyces variotii, and Candida albicans . In the in vitro study, four organisms survived in flasks of sterile saline for the 2 weeks in which serial cultures were performed: K . pneumoniae, C . albicans, A . fumigatus, and P . variotii . In the in vivo study, 61 white rabbits (122 implants) received both an experimental implant inoculated with one of the test organisms and a control implant containing only sterile saline . They were sacrificed at 1-, 3-, or 6-month scheduled endpoints . None of the control implants containing sterile saline had positive cultures . In contrast, the intraluminal saline was culture positive for 7 of the 10 inoculated organisms after varying lengths of time: S . epidermidis, E . coli, E . cloacae, K . pneumoniae, P . aeruginosa, A . fumigatus, and P . variotii . Samples of capsular tissue also were cultured . Of the 122 capsular tissue specimens, 21 (17 percent) had positive cultures and surrounded both inoculated and sterile implants . In most instances, capsules that were culture positive contained an organism different from the one that had been inoculated in the group . In only 3 cases was the same organism cultured from both the periprosthetic tissue and the intraluminal saline, and these may represent instances of the inoculated organism migrating through the implants filler valves . The data show that several types of bacteria (particularly gram-negative species) and fungi can grow and reproduce in a restricted saline environment for extended periods of time. Cancer Res, 1997 Jul 1, 57(13), 2569 - 74 Protective immunity induced by tumor vaccines requires interaction between CD40 and its ligand, CD154; Mackey MF et al.; Interactions between CD40 and its ligand, CD154 (CD40L, gp39), have been shown to play a central role in the regulation of humoral immunity . Recent evidence suggests that this ligand-receptor pair also plays an important role in the induction of cell-mediated immune responses, including those directed against viral pathogens, intracellular parasites, and alloantigens . The contribution of this ligand-receptor pair to the development of protective immunity against syngeneic tumors was evaluated by blocking the in vivo function of CD154 or by studying tumor resistance in mice genetically deficient in CD40 expression (CD40-/-) . In the former case, anti-CD154 monoclonal antibody treatment inhibited the generation of protective immune responses after the administration of three potent tumor vaccines: irradiated MCA 105, MCA 105 admixed with Corynebacterium parvum adjuvant, and irradiated B16 melanoma cells transduced with the gene for granulocyte macrophage colony-stimulating factor . Confirmation of the role of CD40/CD154 interactions in tumor immunity was provided by the overt tumor susceptibility in CD40-deficient mice as compared to that in CD40+/+ mice . In this case, wild-type but not CD40-deficient mice could be readily protected against live TS/A tumor challenge by preimmunization with TS/A admixed with C . parvum . These findings suggest a critical role for CD40/CD154 interactions in the induction of cellular immunity by tumor vaccines and may have important implications for future approaches to cell-based cancer therapies. Vet Microbiol, 1997 Jun 16, 56(3-4), 269 - 76 Oxidation of macrophage membrane cholesterol by intracellular Rhodococcus equi; Linder R et al.; Phagocytic uptake by cultured mouse macrophages (PD388D1) of a virulent strain (ATCC 33701) of Rhodococcus equi producing substantial cholesterol oxidase was accompanied by intracellular survival of the bacteria, and enzymatic oxidation of macrophage membrane cholesterol . A non-virulent strain (4219) lacking cholesterol oxidase was largely eliminated from the macrophages and did not bring about oxidation of membrane cholesterol . When R . equi 33701 was co-phagocytosed with Corynebacterium pseudotuberculosis there was a significant enhancement (10-fold) in the amount of oxidation product (4-cholesten-3-one) generated . R . equi and C . pseudotuberculosis are cooperative partners in the hemolysis of sheep erythrocytes, traceable to the cholesterol oxidase of the former, and phospholipase D of the latter . Results are discussed relative to the role of cooperative cytotoxins in damage to host tissue by bacterial pathogens. FEMS Microbiol Lett, 1997 Jun 15, 151(2), 125 - 30 Acyl phosphatidylglycerol: a major phospholipid of Corynebacterium amycolatum; Yague G et al.; The type strain and several clinical isolates of Corynebacterium amycolatum were examined for lipid composition as a chemotaxonomic character for routine identification . The phospholipid profile was composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides, together with various unidentified compounds . One of them, accounting for 20-29% of total phospholipids, was purified and characterized as acyl phosphatidylglycerol by chromatographic and spectrometric techniques . The acyl substituents on the phosphatidyl moiety were characterized as tetradecanoyl, pentadecanoyl, hexadecenoyl, hexadecanoyl, heptadecenoyl, heptadecanoyl, octadecenoyl (the major one), and octadecanoyl . The acyl group on the polar head (glycerol) was only octadecenoyl . Phospholipid analysis by thin-layer chromatography of a collection of Corynebacterium strains proved that this compound is widely distributed, although it only represents a minor (2-9%) component among mycolic acid-containing species . Acyl phosphatidylglycerol can be considered as a useful chemical marker for the identification of C . amycolatum in addition to the absence of mycolic acids. Commun Dis Intell, 1997 Jun 12, 21(12), 161 - 4 Infections with Corynebacterium diphtheriae--changing epidemiology and clinical manifestations . Report of the third international meeting of the European Laboratory Working Group on Diphtheria (ELWGD), Institute Pasteur, Paris 7-8 June 1996; Gilbert L; A widespread epidemic of diphtheria began in 1990 in the former Soviet Union, in the context of falling immunisation rates and social disruption . Control was impeded by limited diagnostic resources in affected countries (mainly Russia) and there was a risk of spread to neighbouring countries . The European Laboratory Working Group on Diphtheria (ELWGD) was formed to assist in control of the epidemic . The ELWGD is convened by the Public Health Laboratory Service in the United Kingdom and includes 15 laboratories in Europe, one in North America (Centers for Disease Control and Prevention) and the Institute of Clinical Pathology and Medical Research, Sydney . At the group's last annual meeting in Paris, reports were presented on the progress of the epidemic, control strategies and improvements in laboratory diagnosis . The group discussed the increased carriage of, and infection with, nontoxigenic Corynebacterium diphtheriae in countries with high immunisation rates, including the United Kingdom and Australia . They also considered the possible relationship between this increase and the continued diphtheria outbreak in eastern Europe . Preliminary results of molecular typing of toxigenic and nontoxigenic isolates from many parts of the world were presented . It was agreed that further epidemiological investigation is required, using a standardised ribotyping system. Acta Orthop Scand, 1997 Jun, 68(3), 225 - 30 Autotransfusion--bacterial contamination during hip arthroplasty and efficacy of cefuroxime prophylaxis . A randomized controlled study of 40 patients; Wollinsky KH et al.; 40 patients undergoing primary hip arthroplasty, given autologous processed blood transfusion, were randomized a receive no antibiotic prophylaxis (group A, n 20) or cefuroxime (1.5 g single injection; group B, n 20) . Bacterial contamination at various steps in the autotransfusion procedure was assessed in liquid and solid culture media . The operation field and the wound drainage blood were never contaminated either of the groups but some of the suction tips were . Parts of the Vacufix blood collection bags of group A contained bacteria, but none in group B . Processed red blood cell concentrates in both groups showed bacterial growth . Greater blood loss did not increase the contamination rate in general . Isolated bacteria included the species Staphylococcus epidermidis, coagulase-negative staphylococci and Propionibacteria in both groups, but with different cell counts . In addition, Corynebacterium bovis et minutissimum and Moraxelle were identified in group A . In conclusion, autologous blood transfusion was a safe procedure . If contamination occurred, the bacterial count was low, and the bacteria of low pathogenicity . Antibiotic prophylaxis with cefuroxime reduced this contamination of suction tips and collection bags and limited the transfer of autologous blood products. Biosci Biotechnol Biochem, 1997 Jun, 61(6), 960 - 4 Construction of a plasmid carrying both CTP synthetase and a fused gene formed from cholinephosphate cytidylyltransferase and choline kinase genes and its application to industrial CDP-choline production: enzymatic production of CDP-choline from orotic acid (Part II); Fujio T et al.; A new method for enzymatic production of cytidine diphosphate choline (CDP-choline) from orotic acid and choline chloride was developed . To establish an industrial manufacturing process, we constructed a plasmid, pCKG55, which simultaneously expressed in Escherichia coli the three following enzymes; CTP synthetase (encoded by the pyrG gene from E . coli), cholinephosphate cytidylyltransferase (encoded by the CCT gene from Saccharomyces cerevisiae), and choline kinase (encoded by the CKI gene from S . cerevisiae) . CCT and CKI genes on pCKG55 were designed to be expressed as a single CCT/CKI fused protein . This CCT/CKI fused protein retained both activities and the thermal stability of its cholinephosphate cytidylyltransferase activity was nearly the same as the native CCT enzyme . Corynebacterium ammoniagenes KY13505 and E . coli MM294/pCKG55 were cultured in 5-liter jar fermentor independently . Equal volumes of each broth were mixed in a 2-liter jar fermentor, and then the enzymatic reaction was done using 47 mM orotic acid and 60 mM choline chloride as substrates . After 23 h of the reaction at 32 degrees C, 21.5 mM (11 g/liter) of CDP-choline was accumulated. Biosci Biotechnol Biochem, 1997 Jun, 61(6), 956 - 9 Enzymatic production of pyrimidine nucleotides using Corynebacterium ammoniagenes cells and recombinant Escherichia coli cells: enzymatic production of CDP-choline from orotic acid and choline chloride (Part I); Fujio T et al.; Enzymatic production of cytidine diphosphate choline (CDP-choline) using orotic acid and choline chloride as substrates was investigated using a 200-ml beaker as a reaction vessel . When Cornybacterium ammoniagenes KY13505 cells were used as the enzyme source, UMP was accumulated up to 28.6 g/liter (77.6 mM) from orotic acid after 26 h of reaction . In this reaction, UDP and UTP were also accumulated, but CTP, a direct precursor of CDP-choline, was not accumulated sufficiently . Escherichia coli JF646/pMW6 cells, which overproduce CTP synthetase by selfcloning of the pyrG gene, were used together with cells of KY12505 for the enzymatic reaction using orotic acid as a substrate . CTP was produced at 8.95 g/liter (15.1 mM) after 23 h of this reaction . To produce CDP-choline, two additional enzyme activities were needed . E . coli MM294/pUCK3 and MM294/pCC41 cells, which express a choline kinase from Saccharomyces cerevisiae (CKIase; encoded by the CKI gene) and a cholinephosphate cytidylyltransferase from S . cerevisiae (CCTase; encoded by the CCT gene) respectively, were added to this CTP-producing reaction system . After 23 h of the reaction using orotic acid and choline chloride as substrates, 7.7 g/liter (15.1 mM) of CDP-choline was accumulated without addition of ATP or phosphoribosylpyrophosphate (PRPP) . ATP and PRPP required in the CDP-choline forming reaction system are biosynthesized by those cells using glucose as a substrate. Lung Cancer, 1997 Jun, 17 Suppl 1, S121 - 36 Alternatives to chemotherapy and radiotherapy as adjuvant treatment for lung cancer; Shepherd FA; Because adjuvant chemotherapy has resulted in only modest prolongation of survival for patients with lung cancer, investigators have turned to the evaluation of alternative treatment strategies for this patient population . Immunotherapy with Bacillus Calmette Guerin, Corynebacterium parvum, and levamisole has been evaluated in several prospective randomized trials, and no study has shown a statistically significant difference in overall survival . Interferon has been evaluated in three trials of adjuvant therapy after response to chemotherapy for small cell lung cancer . Different interferon preparations were used, but none of the trials showed a significant prolongation of survival . The retinoids have been evaluated as adjuvant treatment after complete resection of stage IN-SCLC . One trial showed a reduction in second primary tumors, and in particular, tumors to tobacco smoking in patients treated with retinyl palmitate . A second trial using 13-cis retinoic acid is ongoing in North America . In the last decade, several inhibitors of angiogenesis have been identified, and they are now beginning to be evaluated in the clinical setting . The National Cancer Institute of Canada Clinical Trials Group and the European Organization for Research and Treatment of Cancer have initiated a study of adjuvant marimastat, a metalloproteinase inhibitor, for patients who have responded to induction chemotherapy for small cell lung cancer . This is the first adjuvant antiangiogenesis factor trial to be initiated for any tumor type . Other investigational agents which are currently undergoing Phase I and Phase II testing include monoclonal antibodies which may inhibit tumour cell growth by binding to growth factors, or which may be conjugated to toxins or chemotherapeutic agents which result in tumour cell death . In the last decade, we have witnessed an explosion in our knowledge and understanding of the regulation of normal and neoplastic cell growth at the molecular level . It remains only speculative at this time as to whether manipulation of abnormal genes in malignant cells will be clinically possible, and whether treatment of this sort may be applied in an adjuvant setting. Microb Pathog, 1997 Jun, 22(6), 343 - 51 Characterization of bacteriophages from tox-containing, non-toxigenic isolates of Corynebacterium diphtheriae; Cianciotto NP et al.; Non-toxigenic strains of Corynebacterium diphtheriae continue to cause disease within immunized populations . A subset of these corynebacteria carry the diphtheria toxin gene but in a cryptic form . To determine whether such strains might contribute to the re-emergence of functional toxin genes, the phages and tox mutations within three clone types were examined . tox-containing, beta-related phages were isolated from two of the strain types . The third isolate appeared to harbour a defective prophage . One of the tox- phages encoded truncated, yet enzymatically-active, forms of diphtheria toxin, suggesting that it had sustained a point mutation within the latter half of its toxin gene . In contrast, the other mutant phage did not elicit the production of either a cross-reacting material or an ADP-ribosylating activity . Complementation tests employing a series of double lysogens confirmed that the mutations responsible for the non-toxigenic phenotype of all of the phages were cis dominant . Given these findings, it is reasonable to hypothesize that tox+ genes can arise within human populations by either homologous recombination between two distinct tox- phages or spontaneous reversion within a single mutant allele. Arch Biochem Biophys, 1997 Jun 1, 342(1), 176 - 81 Folate utilization by monomeric versus heterotetrameric sarcosine oxidases; Wagner MA et al.; There are two types of bacterial sarcosine oxidases . The heterotetrameric enzymes contain subunits ranging in size from about 10 to 100 kDa, noncovalently bound FAD and NAD+, and covalently bound FMN attached to the beta subunit (42-45 kDa) . Monomeric sarcosine oxidases are similar in size to the beta subunit in the heterotetramers and contain covalently bound FAD . Formaldehyde formation during sarcosine oxidation by several heterotetrameric sarcosine oxidases was suppressed in the presence of 50 microM {6S}-tetrahydrofolate, accompanied by a 25-50% increase in the rate of sarcosine oxidation . In contrast, {6S}-tetrahydrofolate caused only a modest decrease in the rate of formaldehyde production with monomeric sarcosine oxidases (approximately 25%), an effect which was virtually entirely attributable to an accompanying decrease in the rate of sarcosine oxidation . In the presence of 100 microM {6R,S}-tetrahydropteroyltriglutamate {H4Pte(Glu)3}, the heterotetrameric enzymes catalyzed the formation of 5,10-methylenetetrahydropteroyltriglutamate {5,10-CH2-H4Pte(Glu)3} at a rate which was 35-60% faster than the rate of sarcosine oxidation in the absence of folate . An apparent Km value of 3.1 microM was estimated for {6S}-H4Pte(Glu)3 with the heterotetrameric corynebacterial sarcosine oxidase . In contrast, slow formation of 5,10-CH2-H4Pte(glu)3 was detected during sarcosine oxidation with monomeric sarcosine oxidases, attributable to the nonenzymatic reaction of free formaldehyde with H4Pte(Glu)3 . The results show that only the heterotetrameric sarcosine oxidases can use tetrahydrofolates as substrates and, in this regard, they resemble mammalian sarcosine and dimethylglycine dehydrogenases. FEMS Microbiol Lett, 1997 May 15, 150(2), 219 - 24 Corynebacterium lipophiloflavum sp . nov . isolated from a patient with bacterial vaginosis; Funke G et al.; A unique coryneform bacterium was isolated from a patient with bacterial vaginosis . Chemotaxonomical investigations demonstrated that the unknown bacterium belonged to the genus Corynebacterium . The yellow-pigmented, slightly lipophilic, oxidative, urea-hydrolyzing bacterium could be phenotypically readily differentiated from the other members of the genus Corynebacterium . Comparative 16S rRNA gene analysis revealed that the bacterium represented a new subline within the genus Corynebacterium for which the name Corynebacterium lipophiloflavum sp . nov . is proposed . The type strain is CCUG 37336 (DSM 44291). J Am Vet Med Assoc, 1997 May 15, 210(10), 1499 - 502 Association between management practices, dairy herd characteristics, and somatic cell count of bulk tank milk; Wilson DJ et al.; OBJECTIVE: To determine whether particular dairy management practices and herd characteristics were associated with somatic cell count (SCC) of bulk tank milk . DESIGN: Analysis of records . SAMPLE POPULATION: Milk samples collected from 59,435 cows housed in 843 dairy herds between March 1992 and June 1994 . PROCEDURE: Results of bacterial culture of milk samples and data on farm housing, sanitation, milking system, and management were collected . Multiple regression analysis was used to determine sources of variation in bulk tank milk SCC among herds . RESULTS: Prevalence of Streptococcus agalactiae and Staphylococcus aureus mastitis was associated with bulk tank milk SCC . In herds free of S agalactiae mastitis, prevalence of S aureus and Corynebacterium bovis mastitis were important . For herds without S agalactiae mastitis, use of sawdust bedding was associated with a decrease in SCC and a dirty loose housing area was associated with an increase . Increased milk production, repeated mastitis control visits, and use of particular predip compounds were significantly associated with reduced SCC in all herds, regardless of whether any cows in the herd had S agalactiae mastitis . In herds with S agalactiae mastitis, use of iodine (certain concentrations), chlorhexidine, peroxide, or sodium chlorite-lactic acid as a predip was associated with a decrease in SCC . Only use of sodium chlorite-lactic acid predip was significantly associated with a decrease in SCC in herds without S agalactiae mastitis . CLINICAL IMPLICATIONS: Important factors associated with bulk tank milk SCC were prevalence of S agalactiae and S aureus mastitis, careful application of particular predip compounds, avoiding a dirty loose housing area, and use of a service to regularly monitor prevalence of mastitis in the herd. Vet Immunol Immunopathol, 1997 May, 56(3-4), 299 - 310 Nitric oxide production following in vitro stimulation of ovine pulmonary alveolar macrophages; Bogdan JR et al.; Production of inducible nitric oxide (NO) as measured by nitrite in supernatant from ovine pulmonary alveolar macrophage (PAM) cultures was assessed following stimulation of PAM with live cells and supernatants from Corynebacterium pseudotuberculosis and Pasteurella haemolytica cultures; purified bacterial lipopolysaccharide derived from both Escherichia coli and Pasteurella haemolytica alone and in combination with interferon-gamma or lymphocyte conditioned medium; or ovine lentivirus . PAM cultured ex vivo with no further stimulation for 24 h, 48 h or 72 h, produced low concentrations of NO that was not substantially increased following co-culture by the various additives . Assessment of NO production in PAM cultures containing P . haemolytica or supernatant from P . haemolytica cultures was complicated by production of high levels of nitrite in the bacterial cultures . Species differences in inducible NO production may affect the efficacy of clearance of bacterial infections and be responsible for inter-host differences in disease expression following infection by intracellular pathogens. Pathology, 1997 May, 29(2), 231 - 3 Corynebacterium pseudotuberculosis is a cause of human necrotising granulomatous lymphadenitis; Mills AE et al.; Corynebacterium pseudotuberculosis is a well recognised pathogen of farm animals, particularly sheep and goats . Human infection is a rare occurrence . This report describes suppurative lymphadenitis occurring in an adolescent boy who had contact with farm animals . The histological differential diagnosis of suppurative granulomatous lymphadenitis is discussed, and the importance of lymph node culture is stressed. Biosci Biotechnol Biochem, 1997 May, 61(5), 858 - 63 Purification and characterization of a dipeptidyl carboxypeptidase from Pseudomonas sp . WO24; Ogasawara W et al.; A dipeptidyl carboxypeptidase (DCP) activity was detected in cell-free extracts of Pseudomonas sp . WO24 . After purification and characterization the enzyme was found to be homogeneous by SDS-PAGE, and had a molecular mass of 74,000 Da by SDS-PAGE and 72,000 Da by gel filtration, indicating that it is monomeric . The isoelectric point was 5.2 and optimum pH was 6.5-7.0 . It showed a specific activity of 780 mumol/min/mg, which is the highest of the values shown by known enzymes . The enzyme hydrolyzed angiotensin I to angiotensin II and sequentially released Phe-Arg and Ser-Pro from the C-terminus bradykinin . The DCP could not cleave imido-bonds, Gly-Gly bonds, or tripeptides . The enzymatic activity was completely inhibited by 0.001 mM EDTA and 0.1 mM O-phenanthroline, but it was not affected by general serine and cysteine protease inhibitors . Addition of Zn2+ completely restored the original activity of the inactivated DCP treated with EDTA . These results suggest that this enzyme is a zinc metalloprotease . The characteristics of the purified enzyme are slightly different from those of the DCPs from Escherichia coli, Pseudomonas maltophilia, and Corynebacterium equi, and considerably from those of the DCP from Bacillus pumilus. Biosci Biotechnol Biochem, 1997 May, 61(5), 840 - 5 High level expression of XMP aminase in Escherichia coli and its application for the industrial production of 5'-guanylic acid; Fujio T et al.; To improve the efficiency of the enzymatic conversion of 5'-xanthylic acid (XMP) to 5'-guanylic acid (GMP), we attempted to increase the activity of the conversion enzyme, XMP aminase (GMP synthetase) encoded by the guaA gene in Escherichia coli . By connecting the PL promoter of lambda phage, the SD sequence of trpL of E . coli, and ATG, at a suitable position upstream of the guaA gene, we obtained plasmid pPLA66 . Sequencing of the nucleotides of the upstream region of the guaA gene on pPLA66 showed that the C-terminal region of the guaB gene, which encodes IMP dehydrogenase, was conserved and a short peptide consisted of 14 amino acids was coded . E . coli MP347/pPLA66 showed an increase in the activity of approximately 370 times when compared with that of the strain MM294, and the amount of the enzyme protein represented approx . 34% of the total cellular protein . Strain MP347/pPLA66 was cultivated in a 5-liter jar fermentor using a medium which contained mainly corn steep liquor . The culture broth had high XMP aminase activity . In the conversion reaction using mixed broths consisted of 600 ml of XMP-fermentation broth of Corynebacterium ummoniagenes KY13203 and 30 ml of cultured broth of E . coli MP347/pPLA66, a surfactant, Nymeen S-215 and xylene were added to the reaction mixture to make the cell membrane permeable to nucleotides . After 23 h of the reaction, 70 mg/ml (131 mM) of GMP.Na2.7H2O was accumulated from 83 mg/ml (155 mM) of XMP.Na3.7H2O, without addition of ATP . The molar conversion yield was approx . 85% . The facts that the cell membrane was treated to allow nucleotides to permeate and that the conversion reaction proceeded well enough in spite of a small amount of E . coli cells indicate ATP was regenerated from AMP by C . ammoniagenes cells and supplied to E . coli cells . Therefore, it was considered that the coupling reaction between these two kind of strains was established. Ophthalmic Surg Lasers, 1997 May, 28(5), 365 - 9 Positive vitreous cultures from eyes without signs of infectious endophthalmitis; Mames RN et al.; BACKGROUND AND OBJECTIVE: There is little information on the rate of false-positive vitreous cultures, because cultures from presumably sterile vitreous are not routinely taken in clinical practice . The objective of this study was to determine the rate of positive vitreous cultures from patients who have no signs of endophthalmitis . PATIENTS AND METHODS: Aerobic cultures from vitreous biopsies were taken from 36 consecutive eyes in which there was no clinical evidence of endophthalmitis . Effluent collected in cassettes during pars plana vitrectomies was processed and cultured in a standard manner . Balanced salt solution was processed intraoperatively through the vitrector and cultured as a negative control . RESULTS: Positive cultures were obtained in 8 of 36 eyes (22.2%) . Coagulase-negative Staphylococcus and Corynebacteria accounted for 7 of the 9 identified organisms . No organism was grown in more than one medium . None of the patients were treated for endophthalmitis after surgery, and none had signs of intraocular infection . CONCLUSIONS: A substantial number of vitrectomy cultures from effluent specimens grow low-virulence organisms in the absence of clinical signs of endophthalmitis . The absence of inflammation at the time of surgery suggests that these positive cultures are contaminants. Protein Sci, 1997 May, 6(5), 1114 - 8 Comparison of high-resolution structures of the diphtheria toxin repressor in complex with cobalt and zinc at the cation-anion binding site; Pohl E et al.; The diphtheria toxin repressor (DtxR) from Corynebacterium diphtheriae is a divalent-metal activated repressor of chromosomal genes responsible for siderophore-mediated iron-uptake and of a gene on several corynebacteriophages that encodes diphtheria toxin . Even though DtxR is the best characterized iron-dependent repressor to date, numerous key properties of the protein still remain to be explained . One is the role of the cation-anion pair discovered in its first metal-binding site . A second is the reason why zinc exhibits its activating effect only at a concentration 100-fold higher than other divalent cations . In the presently reported 1.85 A resolution Co-DtxR structure at 100K, the sulfate anion in the cation-anion-binding site interacts with three side chains that are all conserved in the entire DtxR family, which points to a possible physiological role of the anion . A comparison of the 1.85 A Cobalt-DtxR structure at 100K and the 2.4 A Zinc-DtxR structure at room temperature revealed no significant differences . Hence, the difference in efficiency of Co2+ and Zn2+ to activate DtxR remains a mystery and might be hidden in the properties of the intriguing second metal-binding site . Our studies do, however, provide a high resolution view of the cationanion-binding site that has most likely evolved to interact not only with a cation but also with the anion in a very precise manner. Arch Microbiol, 1997 May, 167(5), 317 - 24 Temperature-sensitive mutants of Corynebacterium ammoniagenes ATCC 6872 with a defective large subunit of the manganese-containing ribonucleotide reductase; Luo C et al.; Chemical mutagenesis of the nucleotide-producing strain Corynebacterium ammoniagenes ATCC 6872 with N-methyl-N-nitro-N-nitrosoguanidine followed by an enrichment protocol yielded 46 temperature-sensitive (ts) clones . A rapid assay for the allosterically regulated Mn-ribonucleotide reductase (RRase) was developed with nucleotide-permeable cells of C . ammoniagenes in order to screen for possible defects in DNA precursor biosynthesis at elevated temperature . Three mutants (CH 31, CH 32, and CH 33) grew well at 30 degrees C but did not proliferate at 40 degrees C because they did not reduce ribonucleotides to 2'-deoxyribonucleotides . They were designated nrdts (nucleotide reduction defective) . When the cultures were shifted from 30 to 40 degrees C, the nrdts mutants immediately ceased to incorporate radiolabeled nucleic acid precursors into the DNA fraction, while DNA chain elongation was barely affected . Thus, exhaustion of the deoxyribonucleotide pool ultimately inhibited cell division, leading to a filamentous growth morphology . In contrast to the wild-type, all three nrdts mutants displayed a distinctly enhanced sensitivity of ribonucleotide reduction towards hydroxyurea (in permeabilized cells and in vitro) at 30 degrees C . The results from assays for biochemical complementation of heat-inactivated (2 min, 37 degrees C) mutant enzyme with either the small or the large subunit of wild-type Mn-RRase located the mutational defect on the large subunit. Vet Rec, 1997 Apr 19, 140(16), 423 - 7 Corynebacterium pseudotuberculosis infection in Israeli cattle: clinical and epidemiological studies; Yeruham I et al.; Morbidity due to Corynebacterium pseudotuberculosis infection occurred in 29 dairy herds in Israel during 1989 to 1995 . The disease occurred sporadically in 17 of the herds with a morbidity of up to 5 per cent, and was epidemic in 12, with a morbidity of 5 to 35 per cent . Cutaneous abscesses were diagnosed in 609 animals . Young cattle appeared to be less susceptible to the disease than older cows . Beef cattle herds were not affected . The disease appeared in the cutaneous form in 92.5 per cent of cases, the cutaneous and mastitic form in 5.9 per cent and the cutaneous and visceral form in 1.6 per cent . The cutaneous form appeared as deep subcutaneous abscesses on various parts of the body, with granulating ulcers exuding pus and blood . In 10 of the herds, C pseudotuberculosis was isolated from 33 mastitic cows which also had cutaneous lesions . The visceral form of the disease was detected when severely affected animals were slaughtered . In 23 of the herds, the disease occurred during the spring and summer dry season, from March to October; the highest prevalence was in the semi-arid Negev region . In 25 herds, the infection lasted for up to five months . The skin lesions on individual cows healed on average in 23.4 days, after either local or parenteral treatment . No significant difference was observed between the effect of systemic antibiotic treatment and local antiseptic treatment . One hundred and two (16.7 per cent) severely affected animals were culled . There was a decrease in milk production and large increases in somatic cell counts in the 12 herds in which the disease was epidemic . None of the strains of isolated C pseudotuberculosis reduced nitrate. MMWR Morb Mortal Wkly Rep, 1997 Apr 18, 46(15), 330 - 2 Respiratory diphtheria caused by Corynebacterium ulcerans--Terre Haute, Indiana, 1996; Opportunistic lung infection with Corynebacterium pseudodiphtheriticum after lung and heart transplantation; St Vincent's Hospital, Sydney, NSWCorynebacterium pseudodiphtheriticum is usually regarded as a harmless commensal of the skin and mucous membranes . We describe two cases of bronchiolitis and bronchitis after lung and heart transplantation, respectively, in which this organism was strongly implicated as the pathogen. Emerg Infect Dis, 1997 Apr-Jun, 3(2), 145 - 53 Rhodococcus equi and Arcanobacterium haemolyticum: two "coryneform" bacteria increasingly recognized as agents of human infection; Linder R; Rhodococcus equi and Arcanobacterium haemolyticum, formerly classified in the genus Corynebacterium, are members of the loosely defined taxon "coryneform" bacteria . Although they are the etiologic agents of distinct human infections, both organisms are frequently overlooked, which results in missed or delayed diagnoses . R . equi, long known as an important pathogen of immature horses, has become in the past three decades an opportunistic pathogen of severely immunosuppressed humans . Most cases are secondary to HIV infection . When specifically sought in throat swab cultures, A . haemolyticum is found responsible for 0.5% to 2.5% of bacterial pharyngitis, especially among adolescents . These two microorganisms represent a spectrum of disease in humans: from a mild, common illness to a rare life-threatening infection . Each organism elaborates lipid hydrolyzing enzymes (cholesterol oxidase by R . equi and sphingomyelinase D by A . haemolyticum) that are toxic to animals and humans and damaging to mammalian cell membranes . The participation of the cytotoxins in pathogenicity is discussed . Greater awareness of the properties of these two bacteria may promote faster, more accurate diagnoses and better clinical management. J Clin Microbiol, 1997 Apr, 35(4), 1011 - 2 Fatal sepsis caused by Corynebacterium amycolatum in a premature infant; Berner R et al.; Corynebacterium amycolatum has not been reported as a cause of human infections up to now, but usually the bacterium is misidentified in clinical specimens as Corynebacterium xerosis . We report the first case of neonatal sepsis due to Corynebacterium amycolatum with a fatal outcome in a premature infant. J Med Microbiol, 1997 Apr, 46(4), 340 - 7 Strategy for detection and identification of bacteria based on 16S rRNA genes in suspected cases of Whipple's disease; Dauga C et al.; The 16S ribosomal RNA (rRNA) gene of the phylogenetic subdivision containing gram-positive bacteria with a high G + C content was detected specifically in clinical specimens from patients suspected of having Whipple's disease . The primary structure of 16S rDNA amplified from clinical samples was determined by cloning and sequencing . Two sorts of sequences were identified: one corresponded exactly to the rRNA sequence of Tropheryma whippelii (GenBank accession no . M87484) while the other was related to that of members of the genus Corynebacterium . No sequence related to Mycobacterium spp . or Rhodococcus equi was observed . Exhaustive examination of negative specimens with broad-range eubacterial primers detected one sequence related to Enterobacteriaceae and another related to Enterococcus spp . To speed identification of T . whippelii, a nested amplification method was devised . A first amplification specific for the gram-positive bacteria subdivision was performed, followed by a second amplification with T . whippelii-specific primers . The amplified T . whippelii product was checked by digestion with AvaII, StuI, and PstI endonucleases . These techniques were applied to DNA extracted from seven intestinal biopsy samples, two cerebrospinal fluid samples and one articular fluid from patients suspected of having Whipple's disease . T . whippelii 16S rDNA was found in two of the biopsy samples, one of the cerebrospinal fluid samples and in the articular fluid. J Bacteriol, 1997 Apr, 179(7), 2449 - 51 A Corynebacterium glutamicum gene conferring multidrug resistance in the heterologous host Escherichia coli; Jager W et al.; A chromosomal DNA fragment from the erythromycin-sensitive bacterium Corynebacterium glutamicum ATCC 13032 was shown to mediate resistance against erythromycin, tetracycline, puromycin, and bleomycin in Escherichia coli . Multicopy cloning of the fragment did not cause a resistance phenotype in C . glutamicum . The corresponding gene encodes a hydrophobic protein with 12 potential transmembrane-spanning ex-helical segments showing similarity to drug-H+ antiporters. Ned Tijdschr Geneeskd, 1997 Mar 15, 141(11), 509 - 11 {Protection against diphtheria}; Hoogkamp-Korstanje JA; An epidemic of (lethal) diphtheria emerged a few years ago in the former Soviet Union where the vaccination density was low . In order to prevent an outbreak in the Netherlands after import of diphtheria, the Health Council went into the degree of protection and measures to increase it . At least one-third of the population are protected insufficiently; they have neither been re-vaccinated in time, nor have they built up immunity by circulation of Corynebacterium strains . Education and (re-)vaccination of risk groups . such as persons having been in contact with an index patient, would appear to be adequate measures. Vet Med (Praha), 1997 Mar, 42(3), 71 - 80 {Validation of TKT medium for detection of Streptococcus agalactiae in bulk milk samples}; Benda P et al.; A selective medium for quantitative determination of Streptococcus agalactiae in bulk milk samples was tested . Among the medium bases, Edwards medium (EW) supplemented with 6 to 9% (v/v) of carefully washed sheep erythrocytes was proved to be best (Tab . V) . The production of sphingomyelinasis C (BHE, Tab . II) or D (COREX, Tab . I) as a supplement supporting the formation of specific hemolytic zones in the medium, was tested in four strains of Staphylococcus aureus and seven strains of Corynebacterium pseudotuberculosis . Submersion aerated cultivation was used for enzyme production; optimum cultivation time is about 100 hours (Figs . 1, 2) . To determine the activity of the enzymes produced a screening method was developed applying hemolytic interactions with CAMP factor of Str . agalactiae . Method sensitivity and reproducibility are sufficiently high for the purposes of observation (Tab . III) . The produced enzymes did not show any changes in their activity over the whole period of storage under all conditions of storage (32 days/7 degrees C, 48 weeks/-18 degrees C, lyophilizate 48 weeks/20 degrees C) . Sphingomyelinasis C is more resistant to heat denaturation, not losing its activity even after being warmed to 85 degrees C for 5 min; sphingomyelinasis D is sensitive to temperatures higher than 50 degrees C (Fig . 3) . COREX was selected (sphingomyelinasis D), produced by the strain CNTCC 18/62 of C . pseudotuberculosis . The enzyme was applied either in raw filtrate of the medium (sterilization by repeated membrane filtration) or in prepurified form obtained by cold acetone precipitation, providing the same results . The medium is highly specific and likely enough selective for Str . agalactiae, unlike BHE it does not provide any positive results with UBERIS-factor+strains of Str . uberis (Tab . V) . Optimum cultivation time is 18 hours; prolongation of cultivation for more than 24 hours brings about the risk of falsely positive readings (Tab . VI) in the strains of accompanying microflora (Morganella morganii ssp . morganii, delta-hemolytic staphylococci). Kansenshogaku Zasshi, 1997 Mar, 71(3), 236 - 40 {Nasopharyngeal flora and carriage rates of Haemophilus influenzae type b of healthy infants}; Asahi E et al.; The present study investigates the nasopharyngeal flora and defines the carriage rates of Haemophilus influenzae type b of 144 healthy infants who visited the well-baby clinic . 224 nasopharyngeal swabs were obtained (44 swabs from 1-month-old infants, 96 swabs from 4 to 5-month-old infants and 84 swabs from 7 to 8-month-old infants), and 440 organisms were isolated . The most frequently isolated organism was Staphylococcus aureus (45.5%) followed by Corynebacterium (38.6%) from 1-month-old infants, Corynebacterium (55.2%) followed by S . aureus (32.3%) and Streptococcus pneumoniae (25.0%) from 4 to 5-month-old infants, Corynebacterium (59.5%) followed by S . pneumoniae (26.2%) from 7 to 8-month-old infants . Twenty-four strains of H . influenzae were isolated from 21 infants, being 1 strain from 1-month-old infant, 9 strains from 4 to 5-month-old infants, 14 strains from 7 to 8-month-old infants . Four type b strains were isolated from 3 infants, being 1 strain from 1-month-old infant, none from 4 to 5-month-old infant, 3 strains from 7 to 8-month-old infants . One of the three 7 to 8-month old infants also carried Hib when he was at the age of 1 month old . The nasopharyngeal carriage rates of H . influenzae of infants were 2.3% in 1-month-old infants, 9.4% in 4 to 5-month-old infants, 16.7% in 7 to 8-month-old infants and 10.7% in total . The carriage rates of Hib were 2.3% in 1-month-old infants, 0% in 4 to 5-month-old infant, 3.6% in 7 to 8-month-old infants and 1.8% in total. Transfusion, 1997 Mar, 37(3), 259 - 63 A retrospective analysis of microbial contaminants in outdated random-donor platelets from multiple sites; Leiby DA et al.; BACKGROUND: Platelet components contaminated with bacteria are an important source of transfusion-associated bacterial sepsis . Estimates of contamination rates vary widely (0-10%) and are highly controversial . The present study, designed with stringent testing regimens, retrospectively determined the prevalence of microbial contaminants in platelets from four collection regions . STUDY DESIGN AND METHODS: During a 9-month period, outdated platelet units were assayed by spreading aliquots from the unit, and from thioglycollate broth medium inoculated with part of the unit, onto 5-percent sheep blood agar media . Cultures were examined after 72-hour incubation at 37 degrees C, and, if bacterial growth was present, the assay processes were repeated with fresh inocula . Units were considered contaminated only if repeatedly positive . RESULTS: Four (0.08%) of 4995 units sampled were contaminated, two with Corynebacterium sp . and one each with Propionibacterium acnes and Aspergillus terreus . Contaminants were present at low, subclinical levels and were detected only after amplification in thioglycollate . The contaminated units were cultured 1, 2, 3, and 7 days after expiration . CONCLUSION: Contamination rates were low and did not vary by region . The identification of A . terreus suggests the role that transfusion may play in transmitting fungal infections should be reassessed . The persistent detection of contaminated platelet units supports the need for a test to detect clinically relevant levels of microbial contaminants in blood components. Semin Respir Infect, 1997 Mar, 12(1), 57 - 60 Rhodococcus equi pneumonia; Johnson DH et al.; Corynebacterium equi is a pleomorphic gram-positive rod that was first isolated in 1923 by Magnusson, and is the cause of suppurative broncho-pneumonia in foals . The organism, now know as Rhodococcus equi, is ubiquitous in nature and is increasingly recognized as pathogenic, particularly in the immunocompromised population . Cavitary pneumonia clinically resembling Mycobacterium tuberculosis is the most common manifestation of R equi disease, but extrapulmonary infections are seen as well . Long term treatment with intracellularly active antimicrobial agents is required. Eur J Pediatr, 1997 Mar, 156(3), 207 - 8 Bacterial tracheitis caused by Corynebacterium diphtheriae; Berner R et al.; Diphtheria has become a rare disease in Germany . We report on an unimmunized 3.5-year-old German girl with a 7-day history of respiratory distress and fever, presenting a clinical picture mimicking typical bacterial tracheitis without pharyngeal and laryngeal manifestation . Diagnosis of diphtheria was not made until culture of tracheal secretions yielded growth of a toxigenic strain of Corynebacterium diphtheriae . The patient died from toxic cardiac failure despite treatment with diphtheria antitoxin . This is the second reported case of isolated bacterial tracheitis caused by Corynebacterium diphtheriae . CONCLUSION: The observation of a lethal course of diphtheric tracheitis emphasizes the paramount importance of immunization against diseases like diphtheria. J Bacteriol, 1997 Mar, 179(5), 1525 - 32 Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number; Nesvera J et al.; The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined . DNA sequence analysis revealed four putative coding regions (open reading frame A {ORFA}, ORFA2, ORFB, and ORFC) . ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C . glutamicum) and pNG2 (from Corynebacterium diphtheriae) . This function was confirmed by deletion mapping of the minimal replicon of pGA1 (1.7 kb) which contains only ORFC . Deletion derivatives of pGA1 devoid of ORFA exhibited significant decreases in the copy number in C . glutamicum cells and displayed segregational instability . Introduction of ORFA in trans into the cells harboring these deletion plasmids dramatically increased their copy number and segregational stability . The ORFA gene product thus positively influences plasmid copy number . This is the first report on such activity associated with a nonintegrating bacterial plasmid . The related plasmids pGA1, pSR1, and pNG2 lacking significant homology with any other plasmid seem to be representatives of a new group of plasmids replicating in the rolling-circle mode. Anal Biochem, 1997 Feb 15, 245(2), 196 - 202 Accurate determination of 13C enrichments in nonprotonated carbon atoms of isotopically enriched amino acids by 1H nuclear magnetic resonance; Wendisch VF et al.; A method for the accurate determination of 13C enrichments in nonprotonated carbon atoms of organic compounds that makes use of unresolved 13C satellites of proton(s) bonded to the vicinal carbon atom was developed . Using glutamate as a model molecule, this 1H nuclear magnetic resonance (NMR) inverse spin-echo difference spectroscopy method was calibrated for inversion efficiency and relaxation effects which were then shown to cause only a minor loss of the measured 13C satellite amplitude (2% for glutamate C-1 and 7% for glutamate C-5) . The determination of 13C enrichments in nonprotonated glutamate carbon atoms by this method was shown to be more precise than 13C NMR . As a first application, a {5-13C}glucose labeling experiment with Corynebacterium glutamicum ASK1 was performed . The labeling patterns of glutamate and arginine extracted from cellular protein were determined using the newly developed method and standard 1H NMR with and without broadband 13C decoupling . Determination of the 13C enrichment in C-5 of glutamate and arginine, respectively, by the two methods showed good agreement . From the deduced labeling pattern of 2-oxoglutarate, an in vivo carbon flux distribution within the central metabolism of C . glutamicum ASK1 was calculated . Thus, the relative flux toward oxaloacetate via the tricarboxylic acid cycle enzyme malate dehydrogenase was determined as 45%, whereas that via anaplerotic C3 carboxylation was determined as 55%. J Ind Microbiol Biotechnol, 1997 Feb-Mar, 18(2-3), 140 - 51 Microbial adaptation to hydrogen peroxide and biodegradation of aromatic hydrocarbons; Fiorenza S et al.; This research investigated microbial responses to bioremediation with hydrogen peroxide (H2O2) as a supplemental oxygen source . Columns containing aquifer material from Traverse City, MI, USA, were continuously supplied with benzene, toluene, ethylbenzene, o-xylene and m-xylene (BTEX) and H2O2 in increasing concentration . The microbial responses studied were changes in microbial numbers, community structure, degradative ability, and activity of catalase and superoxide dismutase (SOD) . Both adaptation to H2O2 and stress-related consequences were observed . Adaptation to H2O2 was demonstrated by increased catalase and SOD activity during the course of the experiment . The microbial community in the untreated aquifer material used in the columns consisted primarily of Corynebacterium sp and Pseudomonas fluorescens . Following amendment with 500 mg L-1 H2O2, the column inlet was dominated by P . fluorescens with few Corynebacterium sp present; Xanthomonas maltophilia dominated the middle and outlet sections . Dimethyl phenols detected in the effluent of two of the biologically active columns were probably metabolic products . The ratio of oxygen to BTEX mass consumed was approximately 0.3 before H2O2 addition, 0.7 following 10 mg L-1 H2O2 supplementation, and 2.6 over the course of the experiment . Abiotic decomposition H2O2 was observed in a sterile column and impeded flow at a feed concentration of 500 mg L-1 H2O2 . Increasing the BTEX concentration supplied to the biologically active columns eliminated flow disruptions by satisfying the carbon and energy demand of the oxygen evolved by increasing catalase activity. Clin Infect Dis, 1997 Feb, 24(2), 185 - 91 Human lymphadenitis due to Corynebacterium pseudotuberculosis: report of ten cases from Australia and review; Peel MM et al.; Corynebacterium pseudotuberculosis commonly causes caseous lymphadenitis in Australian sheep . We describe 10 cases of human lymphadenitis due to C . pseudotuberculosis; in all cases, isolates were submitted to a reference laboratory in Victoria, Australia . Most of the patients were occupationally exposed to sheep . We also review the 12 previously published cases of this infection, most of which were reported from Australia . No patient had any underlying disease or predisposing condition . Surgical excision of the affected lymph glands is the mainstay of management, and antibiotic therapy is supplementary treatment . Diagnosis was delayed for some patients, and some patients had a protracted or recurrent clinical course and/or a slow recovery . These 10 additional cases from one Australian state indicate that human lymphadenitis caused by C . pseudotuberculosis has not been as rare as the number of published reports indicates, at least in Australia . However, the increasing use of a vaccine against caseous lymphadenitis in sheep in Australia should result in the decreasing human incidence of this zoonosis. Hinyokika Kiyo, 1997 Feb, 43(2), 115 - 21 {An experimental study of infection stone formation by Corynebacterium species}; Arai Y et al.; The relationship between infection stone and Corynebacterium species was investigated in vitro and in vivo . Urease activity of urease-splitting Corynebacterium species was evaluated by 2 methods; an increase in pH of human urine after inoculation of Corynebacterium species and direct measurement of urease activity of 10(7) CFU organisms from amounts of ammonia by indophenol method . Formation of infection bladder stone was induced in male Wistar rats by implanting a zinc disc and inoculating 10(6) CFU organisms surgically into the bladder . Urine was alkalinized by the inoculation of Corynebacterium renale, C . pilosum and group D2 Corynebacterium . C . renale and C . pilosum had strong urease activity, and group D2 Corynebacterium had moderate activity . C . pseudodiphtheriticum did not produce the elevation of urinary pH and had little urease activity . Infection stones were formed in 100% of rats by inoculation of C . renale and C . pilosum and 88% of rats by group D2 . Urinary pH was elevated in all inoculated rats . In conclusion, C . renale, C . pilosum and group D2 Corynebacterium may play a role in formation of infection stones. Mol Microbiol, 1997 Feb, 23(3), 483 - 92 The S-layer protein of Corynebacterium glutamicum is anchored to the cell wall by its C-terminal hydrophobic domain; Chami M et al.; PS2 is the S-layer protein of Corynebacterium glutamicum . The S-layer may be detached from the cell as organized sheets by detergents at room temperature . The solubilization of PS2 in the form of monomers requires detergent treatment at high temperature (70 degrees C), conditions under which the protein is denatured . Treatment of the cells with proteinase K or trypsin results in the detachment of the organized S-layer, which remains organized . Because we show that trypsin cleaves the C-terminal part of the protein, we conclude that this domain is involved in the association of the S-layer to the cell but is not essential in the interaction between individual PS2 proteins within the S-layer . A modified form of PS2, deleted of its C-terminal hydrophobic sequence, was constructed . The protein is almost unable to form an organized S-layer and is mainly released into the medium . We suggest that PS2 is anchored via its C-terminal hydrophobic sequence to a hydrophobic layer of the wall of the bacterium located some distance above the cytoplasmic membrane. J Bacteriol, 1997 Feb, 179(3), 838 - 45 Utilization of host iron sources by Corynebacterium diphtheriae: identification of a gene whose product is homologous to eukaryotic heme oxygenases and is required for acquisition of iron from heme and hemoglobin; Schmitt MP; Corynebacterium diphtheriae was examined for the ability to utilize various host compounds as iron sources . C . diphtheriae C7(-) acquired iron from heme, hemoglobin, and transferrin . A siderophore uptake mutant of strain C7 was unable to utilize transferrin but was unaffected in acquisition of iron from heme and hemoglobin, which suggests that C . diphtheriae possesses a novel mechanism for utilizing heme and hemoglobin as iron sources . Mutants of C . diphtheriae and Corynebacterium ulcerans that are defective in acquiring iron from heme and hemoglobin were isolated following chemical mutagenesis and streptonigrin enrichment . A recombinant clone, pCD293, obtained from a C7(-) genomic plasmid library complemented several of the C . ulcerans mutants and three of the C . diphtheriae mutants . The nucleotide sequence of the gene (hmuO) required for complementation was determined and shown to encode a protein with a predicted mass of 24,123 Da . Sequence analysis revealed that HmuO has 33% identity and 70% similarity with the human heme oxygenase enzyme HO-1 . Heme oxygenases, which have been well characterized in eukaryotes but have not been identified in prokaryotes, are involved in the oxidation of heme and subsequent release of iron from the heme moiety . It is proposed that the HmuO protein is essential for the utilization of heme as an iron source by C . diphtheriae and that the heme oxygenase activity of HmuO is involved in the release of iron from heme . This is the first report of a bacterial gene whose product has homology to heme oxygenases. J Clin Microbiol, 1997 Feb, 35(2), 495 - 8 A modified Elek test for detection of toxigenic corynebacteria in the diagnostic laboratory; Engler KH et al.; The detection of toxigenicity among Corynebacterium diphtheriae and Corynebacterium ulcerans strains is the most important test for the microbiological diagnosis of diphtheria . Difficulties with current methods, in particular the Elek test, are well documented . We therefore describe a modified Elek test which provides an accurate result after only 16 h of incubation, in contrast to 48 h for the conventional test. J Clin Microbiol, 1997 Feb, 35(2), 441 - 5 Clinical and molecular study of Corynebacterium diphtheriae systemic infections in France . Coryne Study Group; Patey O et al.; Diphtheria is a disease with a long history that almost completely disappeared from developed countries . In addition, until 1987, systemic infections involving Corynebacterium diphtheriae were rare . However, in 1990, an epidemic occurred in Russia . These two circumstances have provided the stimulus to gain insight into the situation in France . In fact, between 1987 and 1993, a total of 59 C . diphtheriae strains were isolated . Epidemiological data were collected for patients from whom 40 strains were isolated from normally sterile sites, including 34 from blood cultures, and half of the bacteremic patients developed endocarditis . Osteoarticular involvement was noted in 11 of these 40 patients, including 5 bacteremic patients . The fatality rate following bacteremia was 36%, despite specific antibiotic treatment (beta-lactams and aminoglycosides) . The mean age of the participants was 38 years, with half of the patients subsisting under low socioeconomic conditions and suffering from homelessness or alcoholism . Apparently, the skin turned out to be the major route of transmission in this reemerging disease . Eighty-eight percent of the isolates belonged to the C . diphtheriae biotype mitis . These were found predominantly in the Paris area, and most were of the same ribotype . Those isolates originating from the overseas territories (Guyana and New Caledonia) belonged to C . diphtheriae biotype gravis . No strains were positive for the tox gene by PCR . This study attests to the persistent circulation in France of C . diphtheriae in the form of systemic infections . The matter is especially significant since these strains are nontoxigenic and are of a unique ribotype . The strains are, however, sensitive to most antibiotics, although 20% are rifampin resistant. Cas Lek Cesk, 1997 Jan 22, 136(2), 51 - 3 {Review of 105 cases of isolation of Rhodococcus equi in humans}; Votava M et al.; BACKGROUND: Rhodococcus equi (formerly Corynebacterium) has been long considered an exclusively zoopathogenic microbe causing mainly granulomatous pneumonias and lung abscesses in young foals . The aim of this paper was to analyse substantial features of R, equi infections hitherto reported in man . METHODS AND RESULTS: MEDLINE database was searched for relevant reports . When the original source was not obtained the data from reviews were employed . Together, 105 cases of R, equi infection in man were reported . Median age was 35 years with a range from 9 months to 83 years . The male: female ratio was 3.3 . Lungs were involved in 72 cases (69%), extrapulmonary abscesses as the only symptom of infection were described in 9 cases, septic state in 7 cases . Clinical outcome was known in 98 cases, being fatal in 41 (42%) . Therapy was mentioned in 70 reports, the most often used drugs being erythromycin (30 cases, 12 deaths), rifampicin (19 cases, 7 deaths) and vancomycin (18 cases, 6 deaths) . R . equi was isolated from the sputum of 69% patients with the pulmonary involvement . Blood cultures were positive in 35% of cases . Out of total, 49% persons were HIV positive . Median age for HIV positive patients was 32 years with a range from 18 to 71 years, for HIV negative patients 52 years with a range from 9 months to 83 years . There were 97% males in the HIV positive group in contrast to 59% in the HIV negative group (p < 0.01) . Lungs were involved in 90% of HIV positive and 48% of HIV negative cases (p < 0.01) . Extrapulmonary abscesses as the only sign of infection were seen in 2% of HIV positive persons and in 15% of HIV negative ones (p < 0.05) . Outcome was fatal for 60% of the HIV positive hosts and for 28% of the HIV negative individuals (p < 0.01) . R . equi was isolated from the sputum of 80% pneumonic HIV positive patients and of 50% of pneumonic patients without HIV infection (p < 0.05) . R . equi was detected in the blood of 67% of HIV positive patients and of 33% of HIV negative ones (p < 0.01) . CONCLUSIONS: The analysis of published reports shows that whereas R . equi causes mainly pneumonia in persons with HIV infection, in HIV negative individuals extrapulmonary manifestations slightly prevail, most often abscesses, sepsis, eye involvement and wound infections. Mikrobiol Z, 1997 Jan-Feb, 59(1), 65 - 70 {The toxin-containing filtrates from the culture broths of clinical strains of Corynebacterium diphtheriae}; Shchetynina VM et al.; Composition as well as toxicity and antigenicity of toxin-including filtrates of 6 circulating strains of Corynebacterium diphtheriae, isolated in 1992-1994 in the city of Kharkiv have been studied as compared with production strain PW-8 . It is shown that toxicity of filtrates of circulating strains was analogous of toxicity of the strain PW-8 (100 dlm) or lower (45 dlm) . Antigenicity of four strains which was expressed in if units proved to be higher than that of the production strains and it was considerably higher in two strains isolated from adults who died from diphtheria . The methods of gel-filtration and immunoprecipitation were used to establish differences between strains in their capacity to preferential accumulation of different molecular forms of toxin under stationary cultivation. Mikrobiol Z, 1997 Jan-Feb, 59(1), 53 - 60 {The action of bacterial antagonists on the aerobic microflora of human skin}; Klymniuk SI et al.; The authors have investigated antagonistic action of museum strains-antagonists Bacillus subtilis 2896, B . licheniformis 254 and Escherichia coli M-17 against 698 strains of staphylococci, 518 micrococci, 315 corynebacteria, 74 aerobic bacilli and 26 enterobacteria isolated from the surface of stomach and skin of 102 healthy people of different sex . The investigations have shown that B . subtilis 2896 and E . coli M-17 were most active: they inhibited the growth of micrococci, corynebacteria, bacilli, enterobacteria and pseudomonads . Bacteria B . licheniformis have manifested the less antagonistic effect of representatives of skin microbiocenoses . These are B . subtilis and E . coli M-17 that are recommended for clinical testing with the purpose of correction of skin microbiocenoses and for preventing from the development of pyo-septic complications of skin. J Drug Target, 1997, 4(5), 303 - 10 Targeting naproxen to non-parenchymal liver cells protects against endotoxin induced liver damage; Lebbe C et al.; Non-steroidal anti-inflammatory drugs (NSAID's) could be of value in the treatment of liver disease; however, their use in this situation is limited by renal side effects . Therefore, we explored whether naproxen covalently bound to human serum albumin NAP-HSA) was able to reduce toxicity in an acute model of liver disease induced by endotoxin in rats pretreated with Corynebacterium parvum . In the isolated perfused liver of such animals endotoxin induced cholestasis (0.62 +/- 0.05 vs . 0.24 +/- 0.09 microliter.min-1.g liver-1; p < 0.05), increased vascular resistance (11300 +/- 400 vs . 311000 +/- 2000 dyn.s.cm-5; p < 0.05) and alanine aminotransferase release (22 +/- 9 vs . 149 +/- IU/l; p < 0.05) . At the highest dose tested (22 mg/kg, corresponding to 6.0 mumoles naproxen), NAP-HSA normalized ALT release (21 +/- 10 IU/l: p < 0.05) while an equimolar amount of non-targeted naproxen was only partially effective (56 +/- 19 IU/l) . A conventional dose of naproxen similarly prevented transaminase release . Cholestasis and increased vascular resistance were also prevented by NAP-HSA . Drug targeting by linking drugs to proteins is a potentially useful approach to maximizing drug effect while minimizing adverse events; this could be particularly useful for compounds with potentially serious adverse effects in patients with chronic liver disease such as the nonsteroidal anti-inflammatory agents used in the present study. Emerg Infect Dis, 1997 Jan-Mar, 3(1), 65 - 8 Exudative pharyngitis possibly due to Corynebacterium pseudodiphtheriticum, a new challenge in the differential diagnosis of diphtheria; Izurieta HS et al.; Corynebacterium pseudodiphtheriticum has rarely been reported to cause disease in humans, despite its common presence in the flora of the upper respiratory tract . We report here a case of exudative pharyngitis with pseudomembrane possibly caused by C . pseudodiphtheriticum in a 4-year-old girl . The case initially triggered clinical and laboratory suspicion of diphtheria . Because C . pseudodiphtheriticum can be easily confused with Corynebacterium diphtheriae in Gram stain, clarification of its role in the pathogenesis of exudative pharyngitis in otherwise healthy persons is of public health importance . Simple and rapid screening tests to differentiate C . pseudodiphtheriticum from C . diphtheriae should be performed to prevent unnecessary concern in the community and unnecessary outbreak control measures. J Dairy Sci, 1997 Jan, 80(1), 75 - 85 Factors affecting susceptibility to intramammary infection and mastitis: an approximate Bayesian analysis; Rodriguez-Zas SL et al.; Susceptibility to IMI and to mastitis in Holstein cows was studied using logistic mixed effects models and an approximate Bayesian analysis . Dichotomous response variables were the presence or absence of IMI, caused by any microorganism . IMI caused by Staphylococcus spp . or Corynebacterium spp., and clinical mastitis caused by any microorganism at specific lactation stages . Data included 619 lactation records from 282 cows . Fixed explanatory variables in the model were period, season and age at calving, lactation number, log-transformed SCC, and a joint effect of age and log SCC . Because random cow effects were assumed to be normally distributed and to have an unknown variance, this parameter was estimated by approximate marginal maximum likelihood . Results from the Bayesian analysis were contrasted with maximum likelihood estimates obtained from a fixed effects logistic model that ignored cow effects . Posterior mode and maximum likelihood estimates of location parameters were similar, although standard errors of the maximum likelihood estimates understated uncertainty . The IMI status during a previous lactation was a poor predictor of IMI status in subsequent lactations, and susceptibility increased as SCC increased . Interlactation (logit scale) repeatability estimates of susceptibility ranged from 0.22 to 0.23 . A Taylor series expansion was used to approximate correlations between lactations on a binary scale . These correlations depended on associated fixed effects and ranged between 0.12 and 0.18, which were lower than correlations using the logit scale. Vet Res, 1997, 28(2), 149 - 63 Cytokine gene expression in sheep following experimental infection with various strains of Corynebacterium pseudotuberculosis differing in virulence; Pepin M et al.; Corynebacterium pseudotuberculosis is the causative agent of caseous lymphadenitis in sheep and goats . This disease is characterized by the development of pyogranulomas in the lymph nodes and lung tissue . To measure the cytokine gene expression in C pseudotuberculosis lesions, sheep were inoculated with two attenuated strains (Tox- and PLD-t) and a wild-type (WT) strain of C pseudotuberculosis and were necropsied at 7 or 28 days post-inoculation . The Tox- strain showed a strong reduction in virulence as assessed by the absence of disseminating lesions in the lymph nodes draining the inoculation site in contrast with the WT strain . The PLD-t strain showed an intermediate reduction in virulence . The two attenuated strains, however, induced the same amount of antibodies and IFN-gamma production as the WT strain . Using a semi-quantitative RT-PCR technique, the expression of inflammatory cytokines was found to be higher in the inoculation site, whereas expression of T-cell associated cytokines was more intense in the draining lymph node . On the whole, the infected sheep produced high levels of cytokines in at least one organ on days 7 or 28 post-inoculation . No significant differences in cytokine gene expression were shown between sheep infected with strains differing in virulence . Higher cytokine expression was measured in sheep with pyogranulomas in the draining lymph nodes as compared to those without, especially for interleukin-1 beta and interleukin-8 . Overall, these results taken together confirmed the attenuation of virulence in Tox- and PLD-t strains of C pseudotuberculosis and showed the important role of PLD in disseminating the bacteria from the inoculation site to the draining lymph nodes . The pathogenesis of ovine caseious lymphadenitis was shown to be associated with production of cytokines at the pyogranuloma level, but the local cytokine patterns associated with different courses of infection were not distinguished. Exp Anim, 1997 Jan, 46(1), 47 - 52 Changes in normal vaginal flora of African green monkeys (Cercopithecus aethiops) during the menstrual cycle; Narushima S et al.; Changes in normal vaginal flora of African green monkeys associated with the estrous cycle were examined by the swab method . Bacteroidaceae, corynebacteria, and streptococci were predominant throughout the whole estrous cycle, although individual differences were great . It was also clear that all bacterial species tended to decrease in the menstrual phase . Lactobacilli, the most predominant bacteria in the normal vagina of humans, were detected only in very low numbers in the African green monkey, suggesting that the normal vagina of the African green monkey has a different ecosystem from that of normal human vaginal flora. J Thorac Cardiovasc Surg, 1997 Jan, 113(1), 121 - 9 The treatment of patients with infected implantable cardioverter-defibrillator systems; O'Nunain S et al.; OBJECTIVE: The purpose of this study was to evaluate the treatment of patients with infected implantable cardioverter-defibrillator systems . METHODS: Retrospective analysis was done of the cases of 21 patients treated for implantable cardioverter-defibrillator infection during an 11-year period . RESULTS: Of 723 cardioverter-defibrillator implantations (550 primary implants, 173 replacements), nine (1.2%) were complicated by early postoperative device-related infections . Late infections developed in two patients 19 and 22 months, respectively, after implantation . Ten other patients were transferred to our institution for treatment of cardioverter-defibrillator infection . The time from implantation to overt infection was 2.2 +/- 1.3 months, excluding the two late infections . The responsible organisms were Staphylococcus aureus (9), Staphylococcus epidermidis (6), Streptococcus hemolyticus (1), gram-negative bacteria (3), Candida albicans (1), and Corynebacterium (1) . All patients were treated with intravenous antibiotic drugs . Total system removal was done in 15 patients and partial removal in 2; in 4, the cardioverter-defibrillator system was not explanted . There were no perioperative deaths . A new implantable cardioverter-defibrillator system was reimplanted in 7 patients after 2 to 6 weeks of antibiotic therapy . Ten patients were treated without reimplantation (2 arrhythmia operation, 8 antiarrhythmic drugs) . Four patients (3 patients without explantation and 1 with partial system removal) were treated with maintenance long-term antibiotic therapy . During a mean follow-up of 21 +/- 2.8 months, no patient had clinical recurrence of infection . One patient treated with antiarrhythmic drugs without system reimplantation died suddenly . CONCLUSIONS: Infections that involve implantable cardioverter-defibrillator systems can be safely managed by removing the entire system with reimplantation after intravenous antibiotic therapy . In selected patients in whom the risk for system explantation is high and anticipated life expectancy is short, long-term antibiotic therapy to suppress low-virulence infections may represent an acceptable alternative. J Leukoc Biol, 1997 Jan, 61(1), 24 - 32 Elevated levels of NO in both unchallenged and LPS-challenged C . parvum-primed mice are attributable to the activity of a cytokine-inducible isoform of iNOS; Smith SR et al.; Elevated levels of nitric oxide (NO2-/NO3-) were detected in the serum of mice 3-7 days after priming with Corynebacterium parvum (Propionibacterium acnes) . The serum NO2-/NO3- response was completely inhibited when C . parvum-primed (C . parrum) mice were treated with N(G)-monomethyl-L-arginine (L-NMMA) or aminoguanidine (AG) on days 6 and 7 post priming . The response was also inhibited when the mice were treated with interleukin-10 (IL-10) and the cytokine was most effective when given in multiple doses beginning on the day of priming . In contrast to L-NMMA and AG, IL-10 had no effect on the serum NO2-/NO3- response when administered to the mice on days 6 and 7 post priming . The inducible isoform of NOS (iNOS) appeared to be responsible for the elevated NO2-/NO3- response in C . parvum mice because iNOS transcripts were readily detected in their livers . Moreover, these transcripts as well as the circulating levels of NO2-/NO3- were dramatically reduced when the mice were treated with anti-tumor necrosis factor alpha (anti-TNF-alpha) or anti-interferon-gamma (anti-IFN-gamma) monoclonal antibodies (mAbs) during the priming interval . There was a modest increase (less than twofold) in the serum NO2-/NO3- response following a lipopolysaccharide (LPS) challenge to C . parvum mice (C . parvum/LPS mice) . LPS had a more dramatic stimulatory effect if the levels of NO2-/NO3- preexisting in C . parvum/LPS mice were reduced by treatment with L-NMMA, AG, or IL-10 before the challenge . Thus the levels of NO2-/NO3- that preexisted in C . parvum/LPS mice appeared to influence their ability to mount a NO2-/NO3- response subsequent to the LPS challenge . The NO2-/NO3- response did not contribute to lethality in C . parvum/LPS mice because anti-TNF-alpha and anti-IFN-gamma mAbs were protective but had no effect on serum NO2-/NO3- levels when administered to mice 24 h before the LPS challenge. FEMS Microbiol Lett, 1997 Jan 1, 146(1), 129 - 34 Presence of F420-dependent glucose-6-phosphate dehydrogenase in Mycobacterium and Nocardia species, but absence from Streptomyces and Corynebacterium species and methanogenic Archaea; Purwantini E et al.; A range of organisms known to contain F420 or to be relatives of mycobacteria were examined for F420-dependent glucose-6-phosphate dehydrogenase (FGD) and NADP-dependent glucose-6-phosphate dehydrogenase (NADP-G6PD) activities . All free-growing Mycobacterium species examined (including a virulent Mycobacterium tuberculosis strain) had FGD activities of 0.014-0.418 mumol min-1 mg protein-1, and NADP-G6PD activities of 0.013-0.636 mumol min-1 mg-1 . Armadillo-grown Mycobacterium leprae had FGD activity of 0.008 mumol min-1 mg-1, but no detectable NADP-G6PD activity . Nocardia species also had FGD activity (0.088-0.154 mumol min-1 mg-1) . Streptomyces and Corynebacterium species had no FGD, but had NADP-G6PD . Methanogenic Archaea had neither activity. Int J Syst Bacteriol, 1997 Jan, 47(1), 127 - 31 A proposal to reclassify Nocardia pinensis Blackall et al . as Skermania piniformis gen . nov., comb . nov; Chun J et al.; The type strain of Nocardia pinensis was the subject of chemotaxonomic and 16S ribosomal DNA sequencing studies . The resultant nucleotide sequence was aligned with the sequences of representatives of the genera Corynebacterium, Dietzia, Gordona, Mycobacterium, Nocardia, Rhodococcus, and Tsukamurella, and phylogenetic trees were generated by using the Fitch-Margoliash, maximum-parsimony, maximum-likelihood, and neighbor-joining methods . It was evident from the phylogenetic analyses that N . pinensis represents a distinct phyletic line that is most closely associated with the Gordona clade . This genealogical evidence, together with chemotaxonomic and phenotypic data derived from this and previous studies, indicates that N . pinensis merits generic status within the family Nocardiaceae . Therefore, we propose that N . pinensis Blackall et al . 1989 be reclassified as Skermania piniformis gen . nov., comb . nov . The type strain of Skermania piniformis cleaved an array of conjugated substrates based on the fluorophores 7-amino-4-methylcoumarin and 4-methylumbelliferone. Int J Syst Bacteriol, 1997 Jan, 47(1), 92 - 6 Corynebacterium coyleae sp . nov., isolated from human clinical specimens; Funke G et al.; Over a 5-year period, six isolates of a previously unknown nonlipophilic coryneform bacterium were isolated from human clinical specimens . The most characteristic phenotypic reactions of these isolates included slow fermentative acid production from glucose but no acid production from maltose and sucrose and a strongly positive CAMP reaction . Chemotaxonomic investigations revealed that meso-diaminopimelic acid and mycolic acids were present, that palmitic, oleic, and stearic acids were the predominant cellular fatty acids, and that the G + C content was 62 to 64 mol%, characteristics which are consistent with assignment to the genus Corynebacterium . Phenotypically, the unknown coryneform bacterium could be readily differentiated from all other Corynebacterium species . A 16S rRNA gene sequence analysis and quantitative DNA-DNA hybridization experiments demonstrated unambiguously that the unknown coryneform bacterium is a member of the genus Corynebacterium and is genotypically distinct from all other members of this genus . Based on phenotypic and genotypic findings, a new species, Corynebacterium coyleae sp.nov., is proposed . The type strain of C coylease is strain DSM 44184 (= CCUG 35014). Am J Vet Res, 1997 Jan, 58(1), 17 - 22 Effect of natural infection with minor pathogens on susceptibility to natural infection with major pathogens in the bovine mammary gland; Lam TJ et al.; OBJECTIVE: To evaluate the effect of natural udder infection with minor pathogens on subsequent natural infection with major pathogens . SAMPLE POPULATION: 7 dairy herds with low bulk milk somatic cell count . PROCEDURE: During a 20-month prospective study, milk samples were collected from diary cows at regular intervals and from quarters with clinical signs of mastitis . Incidence of intramammary infection was calculated in uninfected quarters and in quarters infected with minor pathogens . A within-cow, matched case-control analysis was used to evaluate the effect of minor pathogens on subsequent infection with major pathogens . RESULTS: Quarters infected with minor pathogens had higher somatic cell count than did uninfected quarters . In quarters infected with Corynebacterium bovis, the rate of infection with major pathogens was lower, whereas in quarters infected with coagulase-negative Micrococcaceae, the rate of infection with major pathogens was higher than that in uninfected quarters . From the within-cow comparison, it appeared that, in quarters infected with minor pathogens, infection with major pathogens was significantly lower than that in comparable control quarters not infected with minor pathogens . CONCLUSIONS: Minor pathogens have a protective effect against infection with major pathogens . The protective effect of C bovis against subsequent infection with major pathogens appears to be greater than the effect of coagulase-negative Micrococcaceae. Plant Mol Biol, 1996 Dec, 32(5), 831 - 48 Effects of altered phosphoenolpyruvate carboxylase activities on transgenic C3 plant Solanum tuberosum; Gehlen J et al.; Phosphoenolpyruvate carboxylase (PEPC) genes from Corynebacterium glutamicum (cppc), Escherichia coli (eppc) or Flaveria trinervia (fppc) were transferred to Solanum tuberosum . Plant regenerants producing foreign PEPC were identified by Western blot analysis . Maximum PEPC activities measured in eppc and fppc plants grown in the greenhouse were doubled compared to control plants . For cppc a transgenic plant line could be selected which exhibited a fourfold increase in PEPC activity . In the presence of acetyl-CoA, a known activator of the procaryotic PEPC, a sixfold higher activity level was observed . In cppc plants grown in axenic culture PEPC activities were even higher . There was a 6-fold or 12-fold increase in the PEPC activities compared to the controls measured in the absence or presence of acetyl-CoA, respectively . Comparable results were obtained by transient expression in Nicotiana tabacum protoplasts . PEPC of C . glutamicum (PEPC C.g.) in S . tuberosum leaf extracts displays its characteristic K(m) (PEP) value . Plant growth was examined with plants showing high expression of PEPC and, moreover, with a plant cell line expressing an antisense S . tuberosum (anti-sppc) gene . In axenic culture the growth rate of a cppc plant cell line was appreciably diminished, whereas growth rates of an anti-sppc line were similar or slightly higher than in controls . Malate levels were increased in cppc plants and decreased in antisense plants . There were no significant differences in photosynthetic electron transport or steady state CO2 assimilation between control plants and transformants overexpressing PEPC C.g . or anti-sppc plants . However, a prolonged dark treatment resulted in a delayed induction of photosynthetic electron transport in plants with less PEPC . Rates of CO2 release in the dark determined after a 45 min illumination period at a high proton flux density were considerably enhanced in cppc plants and slightly diminished in anti-sppc plants . When CO2 assimilation rates were corrected for estimated rates of mitochondrial respiration in the light, the electron requirement for CO2 assimilation determined in low CO2 was slightly lower in transformants with higher PEPC, whereas transformants with decreased PEPC exhibited an appreciably elevated electron requirement . The CO2 compensation point remained unchanged in plants (cppc) with high PEPC activity, but might be increased in an antisense plant cell line . Stomatal opening was delayed in antisense plants, but was accelerated in plants overexpressing PEPC C.g . compared to the controls. Mol Microbiol, 1996 Dec, 22(5), 815 - 26 A new type of transporter with a new type of cellular function: L-lysine export from Corynebacterium glutamicum; Vrljic M et al.; We discovered that after deregulation of the L-lysine biosynthesis in Corynebacterium glutamicum, L-lysine accumulated in the cytosol and the efflux properties of this amino acid in mutants used for L-lysine production were altered . In this study we describe the cloning and molecular identification of lysE, which encodes the translocator specifically exporting L-lysine from the cell . The lysE gene product does not display homology to any known transporter . It is only 236 amino acids in size, with the potential to span the membrane six times . The LysE protein was oversynthesized to confirm its deduced M(r) of 25425 Da . A probable regulatory gene, lysG, is localized immediately adjacent to lysE and displays all the typical structural features of an autoregulatory transcriptional regulator of the LysR-type family . L-Lysine export is correlated with lysE expression . A null mutant is unable to excrete L-lysine, whereas with overexpressed lysE, L-lysine is exported at a rate of 3.76 nmol min-1 mg-1 dry weight, which is five times the rate that was obtained with the wild type . A deletion mutant was constructed to search for a natural function of this unique carrier . Surprisingly, growth of this mutant is abolished on a salt medium in the presence of the dipeptide Lys-Ala . The quantification of the intracellular L-lysine concentrations revealed that, in response to peptide addition, there was an accumulation of the exceptionally high concentration of more than 1100 mM L-lysine . These results distinguish LysE as an exporter, which: (i) structurally represents a new type of translocator; (ii) demonstrates that exporters are also present for primary metabolites such as amino acids; and (iii) serves in one physiological function to link import with export activity. Clin Infect Dis, 1996 Dec, 23(6), 1246 - 8 Corynebacterium striatum meningitis: case report and review of an increasingly important Corynebacterium species; Weiss K et al.; For a long time, corynebacteria were considered simple cutaneous contaminants with little potential pathogenicity . Corynebacterium striatum is a known saprophytic cutaneous bacterium; however, in the last decade, this organism has been increasingly recognized as a pathogen . It has been mostly implicated in respiratory tract and blood infections . To our knowledge, we report the first case of meningitis due to C . striatum . Treatment with intravenous vancomycin resulted in therapeutic success . We also thoroughly review all previously reported cases of C . striatum infection . Identification of Corynebacterium species can be difficult because of rapid taxonomic changes, and susceptibility testing for these microorganisms is not yet standardized . However, because of their growing clinical importance, data on these bacteria are accumulating. Infect Immun, 1996 Dec, 64(12), 5047 - 52 Characterization of a novel lipoprotein expressed by Haemophilus ducreyi; Hiltke TJ et al.; Pooled sera from patients with chancroid contain antibodies to a Haemophilus ducreyi antigen with an approximate molecular weight of 28,000 (28K) . Rabbit polyclonal serum that reacts to a 28K protein can be used to detect H . ducreyi in clinical samples . A monoclonal antibody, designated 5C9, bound to a 28K outer membrane protein and to 35 of 35 H . ducreyi isolates with diverse geographic origins and did not bind to many species of the families Pasteurellaceae, Neisseriaceae, and Enterobacteriaceae or to Corynebacterium and Candida species strains . A 5C9-reactive phage was recovered from a genomic library, and the gene encoding the 28K protein was localized to a 626-bp open reading frame, designated hlp, for H . ducreyi lipoprotein . Translation of hlp predicted a 23K polypeptide that contained a lipoprotein processing site . Escherichia coli transformed with a plasmid containing hlp expressed a novel, membrane-associated protein that could be labeled with {3H}palmitic acid . In H . ducreyi, processing of Hlp was inhibited by globomycin . Database searches found no homologies to hlp or to the predicted Hlp amino acid sequence . Restriction enzyme analysis indicated that hlp was conserved among H . ducreyi isolates . Serum samples from patients with chancroid and other genital ulcer diseases and from normal subjects contained antibodies that bound to purified, recombinant Hlp . Although monoclonal antibody 5C9 recognizes a species-specific epitope of a unique H . ducreyi lipoprotein, the presence of serum antibodies to Hlp may not indicate previous infection with H . ducreyi. J Immunol, 1996 Dec 1, 157(11), 5022 - 6 Corynebacterium parvum- and Mycobacterium bovis bacillus Calmette-Guerin-induced granuloma formation is inhibited in TNF receptor I (TNF-RI) knockout mice and by treatment with soluble TNF-RI; Senaldi G et al.; The aim of this study was to examine the role of TNF receptor I (TNF-RI) in the pathogenesis of heat-killed Corynebacterium parvum- and live bacillus Calmette-Guerin (BCG)-induced granulomas . Granuloma formation was analyzed in TNF-RI knockout mice and after treatment with soluble TNF-RI (sTNF-RI) . TNF-RI knockout mice injected with C . parvum or BCG developed fewer and smaller granulomas than wild-type control mice . Mice treated with sTNF-RI from days 7 to 13 after injection of C . parvum or BCG developed fewer and smaller granulomas than saline-treated control mice . Established granulomas regressed in rats treated with sTNF-RI from days 10 to 13 after injection of C . parvum . In conclusion, TNF signaling via TNF-RI contributes to the pathogenesis of C . parvum- and BCG-induced granulomas . sTNF-RI inhibits the development of granulomas and can cause the regression of established granulomas. J Infect Dis, 1996 Dec, 174(6), 1380 - 3 Vaccination with alum-precipitated recombinant Ancylostoma-secreted protein 1 protects mice against challenge infections with infective hookworm (Ancylostoma caninum) larvae; Ghosh K et al.; Ancylostoma-secreted protein 1 (ASP-1) is the major protein secreted by infective hookworm larvae (Ancylostoma caninum) . The Escherichia coli-expressed recombinant protein was evaluated as a vaccine antigen in a mouse model of ancylostomiasis . A . caninum larvae migrate through mouse lungs, with maximal migration occurring 48-54 h after oral infection . Quantitative larval recovery from the lungs at this time was used as an end point for vaccine evaluation . All mice developed antibodies to recombinant ASP-1 (rASP-1) after immunization and boosting with the alum-precipitated protein . The immunized mice had their worm burden reduced 79% (P < .0001) compared with controls . Immunization with rASP-1 in the presence of Corynebacterium parvum adjuvant also showed a vaccine effect (63% protection; P < .0001) . The possibility that this protective effect resulted from delayed larval lung entry was excluded . rASP-1 offers promise as a hookworm vaccine antigen. Ned Tijdschr Geneeskd, 1996 Nov 30, 140(48), 2414 - 6 {A tropical ulcer; cutaneous diphtheria}; Werdmuller BF et al.; A 43-year-old woman, born in the Netherlands, developed ulcers on her left foot during a holiday in Gambia and Senegal . She had been bitten bij insects . The ulcers were caused by Corynebacterium diphtheriae . The patient was treated with antibiotics and recovered fully . A grey pseudomembrane covering the ulcer is a characteristic feature of cutaneous diphtheria . The treatment is with antibiotics and, after toxin tests have indicated that the bacterium is toxigenic, with antitoxin . In some cases screening of social contacts is advised. J Biol Chem, 1996 Nov 22, 271(47), 29545 - 51 Mycolic acid structure determines the fluidity of the mycobacterial cell wall; Liu J et al.; The low permeability of the mycobacterial cell wall is thought to contribute to the well known resistance of mycobacteria to antibiotics and chemotherapeutic agents . We have used differential scanning calorimetry to demonstrate that the high temperature phase transition observed in purified cell walls, usually in the 60-70 degrees C range, suggestive of a lipid environment of extremely low fluidity, can also be observed in whole organisms and in cell walls from which much of the free lipids was removed by extraction with Triton X-114 . A survey of seven mycobacterial species demonstrated that this high temperature transition was a general property of these organisms . Cell walls isolated from two Corynebacterium species, which contain much shorter corynemycolic acids, displayed a much lower temperature transition, suggesting that the transition temperature was directly correlated to the length of mycolic acid . Methyl esters of mycolic acids were found to have a phase transition temperature that was linearly related to the amount of trans-mycolate . Both Mycobacterium avium and M . smegmatis responded to increasing growth temperature by increasing the proportion of trans-mycolate and displaying a correspondingly higher melting temperature . Whole cells of M . smegmatis grown at higher temperature allowed a less rapid influx of two lipophilic agents, norfloxacin and chenodeoxycholate . These results provide strong evidence that the nature of mycolic acid plays a crucial role in determining the fluidity and permeability of mycobacterial cell wall. Arkh Patol, 1996 Nov-Dec, 58(6), 28 - 33 {The morphological characteristics of diphtheria in adults under current conditions}; Kadyrova SN et al.; Clinico-anatomical analysis of 112 lethal cases of diphtheria in adults showed a grave course of the disease in chronic alcoholics at the age of 40-49 years . The morphological changes were characterized by a complex of typical changes at the primary focus, intracanalicular and hematogenous dissemination with high bacteremia . The morphology of pneumonia caused by Corynebacterium diphtheria is described for the first time . At the early stages of the disease infectious-toxic degeneration of the myocardium was documented with predominating alteration of the atrioventricular node or contractile myocardium . The above morphological features may be due to peculiarities of patients and decreased subjects as well as possible changes in the pathogen characteristics. Biosci Biotechnol Biochem, 1996 Nov, 60(11), 1826 - 30 Isolation and identification of styrene-degrading Corynebacterium strains, and their styrene metabolism; Itoh N et al.; By supplying styrene in the gas phase as the sole carbon and energy source, styrene-degrading aerobic microorganisms were readily isolated from soil samples . They were identified as Corynebacterium pseudodiphtheriticum and similar species, or Pseudomonas sp . Growth experiments on some aromatic compounds, resting-cell reactions with them, and the measurement of degrading enzyme activities suggest that Corynebacterium sp . AC-5 and St-5 strains metabolize styrene through styrene oxidation into styrene oxide, then convert it into phenylacetaldehyde by a reaction using styrene oxide isomerase, and phenylacetaldehyde is reduced to 2-phenylethanol . The Corynebacterium sp . ST-10 strain did not have styrene oxide isomerase, and metabolized styrene oxide by an unknown enzymatic reaction . Possible metabolism of styrene was proposed for the Corynebacterium strains. Mol Microbiol, 1996 Nov, 22(3), 535 - 44 An ideR mutant of Mycobacterium smegmatis has derepressed siderophore production and an altered oxidative-stress response; Dussurget O et al.; The mycobacterial ideR protein is a homologue of the diphtheria-toxin repressor DtxR . We have previously demonstrated that Mycobacterium tuberculosis ideR, like DtxR, represses transcription of Corynebacterium diphtheriae iron-regulated promoters in vivo and binds to C . diphtheriae operators in a metal-dependent manner in vitro . We show here that ideR mutants of M . smegmatis, constructed by allelic replacement, were defective in their ability to repress siderophore biosynthesis in the presence of iron . They were also more sensitive to hydrogen peroxide and had decreased levels of catalase/peroxidase (KatG) and manganese superoxide dismutase (Mn-SOD) . This indicates that ideR is a negative regulator of siderophore production and is required for the response to superoxide- and hydrogen peroxide stress . We propose that ideR is the mycobacterial counterpart of the Escherichia coli Fur protein, i.e . It is a pleiotropic regulator that couples iron metabolism to the oxidative-stress response. Antimicrob Agents Chemother, 1996 Nov, 40(11), 2671 - 2 Antimicrobial susceptibility pattern of Corynebacterium striatum; Martinez-Martinez L et al.; The in vitro activities of 16 antimicrobial agents against 86 strains of Corynebacterium striatum were evaluated by microdilution using cation-adjusted Mueller-Hinton broth . MICs at which 90% of strains were inhibited were 0.06 microgram/ml for teicoplanin, 1 microgram/ml for vancomycin, 0.03 to 8 micrograms/ml for beta-lactams, 8 micrograms/ml for sparfloxacin, 16 micrograms/ml for ciprofloxacin, 16/304 micrograms/ml for co-trimoxazole (trimethoprim-sulfamethoxazole), 64 micrograms/ml for tetracycline, 128 micrograms/ml for gentamicin, and > 128 micrograms/ml for amikacin, erythromycin, and rifampin. Cell Immunol, 1996 Nov 1, 173(2), 207 - 14 Administration of interleukin-10 at the time of priming protects Corynebacterium parvum-primed mice against LPS- and TNF-alpha-induced lethality; Smith SR et al.; Several laboratories have described the protective effects of interleukin-10 (IL-10) in mouse models of lethal endotoxemia . In most of these experiments, protection was observed in normal mice that were given a lethal dose of LPS . However, we failed to observe protection with IL-10 in LPS-challenged mice that had been primed with Corynebacterium parvum (Proprionibacterium acnes) . We have extended our studies with IL-10 in C . parvum-primed mice and in some cases have observed protection that appears to depend on the strength of the sensitization to C . parvum . When IL-10 was administered to mice at the time of priming, it was particularly effective in blocking sensitization, as evidenced by the inability of treated mice to mount a strong inflammatory cytokine response when subsequently challenged with LPS . Following such treatment with IL-10, C . parvum-primed mice were also protected from a subsequent lethal challenge with rMuTNF-alpha . In addition, the mice were protected against LPS- and TNF-alpha-induced lethality with a single dose of an anti-TNF-alpha or anti-IFN-gamma mAb given at the time of priming . Our results suggest that TNF-alpha and IFN-gamma are produced early after priming with C . parvum and are at least partly responsible for the enhanced sensitivity of the mice to LPS and TNF-alpha . IL-10 affords protection to the mice because of its ability to block the C . parvum-induced TNF-alpha and IFN-gamma responses. Clin Immunol Immunopathol, 1996 Nov, 81(2), 161 - 7 Susceptibility to immunologically mediated fatigue in C57BL/6 versus Balb/c mice; Sheng WS et al.; Proinflammatory cytokines such as interleukin (IL)-1 and tumor necrosis factor (TNF)-alpha have been proposed to play a role in the pathogenesis of fatigue . In the present study we compared the susceptibility of two mouse strains to immunologically induced fatigue . Daily running of two strains of mice, Balb/c and C57BL/ 6, was assessed after a single injection of Corynebacterium parvum antigen (2 mg/mouse) . Spontaneous running activity of each animal was compared to mean running distance prior to injection . To evaluate the involvement of cytokines in fatigue development, C57BL/6 mice were treated with antibodies to specific cytokines at the time of challenge with C . parvum antigen . Also, cytokine mRNA expression was analyzed in the brains of mice at different time periods after immunologic challenge . A significant difference in running activity between the two mice strains was observed after C . parvum antigen inoculation: C57BL/6 mice showing a greater (P < 0.05) reduction in running activity (relative to preinjection levels) and slower recovery to baseline than Balb/c mice . Injection of antibodies specific to either IL-1beta or TNF-alpha did not alter immunologically induced fatigue, suggesting a lack of involvement of these cytokines produced outside of the central nervous system (CNS) . However, increased TNF-alpha and IL-1beta mRNA expression was found in the brains of C57BL/6 compared to that seen in Balb/c mice at 6, 10, and 15 days after C . parvum antigen injection . The elevated CNS cytokine mRNA expression corresponded to development of fatigue . These findings are consistent with the hypothesis that expression of proinflammatory cytokines within the CNS plays a role in the pathogenesis of immunologically mediated fatigue. J Infect Dis, 1996 Nov, 174(5), 1064 - 72 Molecular epidemiology of diphtheria in Russia, 1985-1994; Popovic T et al.; The largest diphtheria outbreak in the developed world since the 1960s began in the Russian federation in 1990 . One hundred fifty-six Corynebacterium diphtheriae strains from throughout Russia, selected for temporal and geographic diversity, were assayed by ribotyping and multilocus enzyme electrophoresis (MEE) . These tests showed significant genetic diversity within the C . diphtheriae species, and ribotyping and MEE data generally correlated well with epidemiologic data . A distinct clonal group of C . diphtheriae isolates (ET 8 complex) emerged in Russia in 1990 as the current outbreak began, and as the outbreak has progressed, these organisms have made up increasingly larger proportions of the strains that are isolated . Furthermore, the main characteristic of the epidemic strains is a specific combination of ET 8 and ribotypes G1 and G4 . This study confirms the epidemiologic utility of the molecular subtyping methods that detected the epidemic clone and addresses the clone's origin and relation to C . diphtheriae from throughout Russia. Curr Microbiol, 1996 Nov, 33(5), 292 - 6 Accelerated Biodegradation of High and Low Concentrations of p-Nitrophenol (PNP) by Bacterial Inoculation in Industrial Wastewater: The Role of Inoculum Size on Acclimation Period Zaidi BR, Imam SH, Greene RV. The effect of inoculum size on the acclimation period and rate and extent of p-nitrophenol (PNP) degradation at high (1-10 mg/L) and low (26 &mgr;g/L) concentrations for two bacteria was determined in defined media as well as industrial wastewater . Increased inoculum size did not affect the acclimation period of either bacterium at high (1-10 mg/L) PNP concentrations . At low PNP concentrations (26 &mgr;g/L), the two bacteria behaved differently . The acclimation period was shortened and both the rate and extent of mineralization of PNP were enhanced by increasing the Corynebacterium sp . inoculum size from 3 x 10(5) to 3 x 10(6) cells/ml . Addition of phosphate or elimination of predators also reduced the acclimation period . Conversely, increasing the inoculum size from 3 x 10(5) to 5 x 10(6) cells/ml of Pseudomonas putida lengthened the acclimation period and reduced both the rate and extent of mineralization . It is suggested that, in a given environment, the success of an introduced species to enhance the degradation of a chemical depends upon (i) concentration of the chemical, (ii) selection of an appropriate microorganism, and (iii) utilization of a suitable inoculum size. Biochemistry, 1996 Oct 22, 35(42), 13540 - 51 Three-dimensional structure of meso-diaminopimelic acid dehydrogenase from Corynebacterium glutamicum; Scapin G et al.; Diaminopimelate dehydrogenase catalyzes the NADPH-dependent reduction of ammonia and L-2-amino-6-ketopimelate to form meso-diaminopimelate, the direct precursor of L-lysine in the bacterial lysine biosynthetic pathway . Since mammals lack this metabolic pathway inhibitors of enzymes in this pathway may be useful as antibiotics or herbicides . Diaminopimelate dehydrogenase catalyzes the only oxidative deamination of an amino acid of D configuration and must additionally distinguish between two chiral amino acid centers on the same symmetric substrate . The Corynebacterium glutamicum enzyme has been cloned, expressed in Escherichia coli, and purified to homogeneity using standard biochemical procedures {Reddy, S . G., Scapin, G., & Blanchard, J . S . (1996) Proteins: Structure, Funct . Genet . 25, 514-516} . The three-dimensional structure of the binary complex of diaminopimelate dehydrogenase with NADP+ has been solved using multiple isomorphous replacement procedures and noncrystallographic symmetry averaging . The resulting model has been refined against 2.2 A diffraction data to a conventional crystallographic R-factor of 17.0% . Diaminopimelate dehydrogenase is a homodimer of structurally not identical subunits . Each subunit is composed of three domains . The N-terminal domain contains a modified dinucleotide binding domain, or Rossman fold (six central beta-strands in a 213456 topology surrounded by five alpha-helices) . The second domain contains two alpha-helices and three beta-strands . This domain is referred to as the dimerization domain, since it is involved in forming the monomer--monomer interface of the dimer . The third or C-terminal domain is composed of six beta-strands and five alpha-helices . The relative position of the N- and C-terminal domain in the two monomers is different, defining an open and a closed conformation that may represent the enzyme's binding and active state, respectively . In both monomers the nucleotide is bound in an extended conformation across the C-terminal portion of the beta-sheet of the Rossman fold, with its C4 facing the C-terminal domain . In the closed conformer two molecules of acetate have been refined in this region, and we postulate that they define the DAP binding site . The structure of diaminopimelate dehydrogenase shows interesting similarities to the structure of glutamate dehydrogenase {Baker, P . J., Britton, K . L., Rice, D . W., Rob, A., & Stillmann, T.J . (1992a) J . Mol . Biol . 228, 662-671} and leucine dehydrogenase {Baker, P.J., Turnbull, A.P., Sedelnikova, S.E., Stillman, T . J., & Rice, D . W . (1995) Structure 3, 693-705} and also resembles the structure of dihydrodipicolinate reductase {Scapin, G., Blanchard, J . S., & Sacchettini, J . C . (1995) Biochemistry 34, 3502-3512}, the enzyme immediately preceding it in the diaminopimelic acid/lysine biosynthetic pathway. Gene, 1996 Oct 10, 175(1-2), 15 - 22 Two new members of the bio B superfamily: cloning, sequencing and expression of bio B genes of Methylobacillus flagellatum and Corynebacterium glutamicum; Serebriiskii IG et al.; Cloning, characterization and expression of the bio B gene of the obligate methylotrophic bacterium, Methylobacillus flagellatum, are reported . A chromosomal fragment containing bio B has been isolated by complementation of a bio B- mutant of M . flagellatum . Nucleotide (nt) sequence analysis of this fragment revealed the presence of an open reading frame of 966 nt identified as bio B, which is the first gene of the M . flagellatum bio cluster . Gene bio B was expressed in Escherichia coli and M . flagellatum, resulting in efficient conversion of dethiobiotin to biotin . The Corynebacterium glutamicum bio B has also been cloned and sequenced . Comparison of the amino acid sequences derived from known bio B genes allowed us to identify four cysteines participating as putative ligands forming the {2Fe-2S} cluster . Genomic organization of the bio biosynthetic genes shows wide diversity in various bacteria . The results of the database screening suggested that bio B proteins belong to a superfamily of proteins, including biotin and lipoate synthases and some proteins with unidentified functions. Postgrad Med J, 1996 Oct, 72(852), 619 - 20 Cutaneous and pharyngeal diphtheria imported from the Indian subcontinent; Hart PE et al.; We report the case of a 32-year-old woman who presented upon returning from India with cutaneous ulcers on the feet and pharyngitis . Microbiological testing showed the causative organism to be a toxigenic strain of Corynebacterium diphtheriae . She was treated successfully with penicillin and diphtheria antitoxin . This case emphasises the importance of maintaining a high index of suspicion for such rare but significant infectious diseases. Vet Microbiol, 1996 Oct, 52(3-4), 313 - 5 Distribution of Corynebacterium renale among apparently healthy rats; Osanai T et al.; We examined the distribution of Corynebacterium renale, a causative agent of urinary calculus, in clinically normal rats at 6 animal facilities in Japan . Swabs of the vulva and vaginal vestibule or prepuce of the rats were cultured for isolation of the organisms . C . renale has been isolated at only one animal facility, where cases of urinary calculus were reported several years ago . In this facility, 32% of female (43/135) and 22% of male (18/82) rats, 4-28 weeks old, were positive for C . renale . In contrast, 92 female and 169 male rats at other facilities without a history of the disease were negative for the organisms. J Chemother, 1996 Oct, 8(5), 387 - 93 Bacteremia and fungemia occurring during antimicrobial prophylaxis with ofloxacin in cancer patients: risk factors, etiology and outcome; Spanik S et al.; The authors analyzed 27 breakthrough bacteremias occurring during ofloxacin prophylaxis in afebrile neutropenia over 7 years in 9989 admissions and 979 bacteremic and fungemic episodes in a National Cancer Center in Bratislava, Slovak Republic . The most frequently isolated organisms in breakthrough bacteremias were gram-positive (71.3%), mainly coagulase-negative staphylococci (41.3%), enterococci (9.2%) and Corynebacteria (9.2%), followed by gram-negative rods-Pseudomonas aeruginosa (13.2%) and Stenotrophomonas maltophilia (9.2%) . The outcome of breakthrough bacteremias during ofloxacin prophylaxis was not associated with the underlying disease, neutropenia, catheter insertion or resistance, but only with multiple risk factors . A higher failure rate was observed in those patients having a catheter infected with a resistant organism and during neutropenia . No patients with Hickman catheter were included in the study . Patients with mixed breakthrough bacteremia due to gram-negative and gram-positive organisms had higher failure rates than those with monomicrobial bacteremia . Catheter extraction and rapid institution of intravenous antibiotics in combination should be administered in breakthrough bacteremia. J Clin Microbiol, 1996 Oct, 34(10), 2625 - 7 Identification of Turicella otitidis isolated from a patient with otorrhea associated with surgery: differentiation from Corynebacterium afermentans and Corynebacterium auris; Renaud FN et al.; Turicella otitidis is a newly described coryneform bacterium isolated from middle ear fluids . We report here on the diagnosis of a strain isolated from otorrhea . The API Coryne system (bioMerieux, Marcy I'Etoile, France) used alone failed to differentiate T . otitidis, Corynebacterium afermentans, and Corynebacterium auris (ANF group) . Biochemical tests such as DNase, enzymatic reactions (API ZYM; bioMerieux), and carbon substrate assimilation tests (Biotype 100; bioMerieux) allow presumptive identification . However, only chemotaxonomy and molecular biology can achieve unequivocal differentiation among these three species. Am J Pathol, 1996 Oct, 149(4), 1303 - 12 Experimental extrinsic allergic alveolitis and pulmonary angiitis induced by intratracheal or intravenous challenge with Corynebacterium parvum in sensitized rats; Yi ES et al.; Extrinsic allergic alveolitis and pulmonary sarcoidosis are granulomatous diseases of the lung for which clinical presentation and anatomic site of granuloma formation differ . Extrinsic allergic alveolitis is caused by inhaled antigens, whereas the nature and source of the inciting antigen in sarcoidosis is unknown . To test the hypothesis that the route via which antigen is introduced to the lung contributes to the clinicopathological presentation of pulmonary granulomatous disease, rats immunized with intravenous (i.v.) Corynebacterium parvum were challenged after 2 weeks with either intratracheal (i.t.) or i.v . C . parvum . The granulomatous inflammation elicited by i.t . challenge predominantly involved alveolar spaces and histologically simulated extrinsic allergic alveolitis . In contrast, the inflammation induced by i.v . challenge was characterized by granulomatous angiitis and interstitial inflammation simulating sarcoidosis . Elevations of leukocyte counts and TNF levels in bronchoalveolar fluid, which reflect inflammation in the intra-alveolar compartment, were much more pronounced after i.t . than after i.v . challenge . Tumor necrosis factor, interleukin-6, CC chemokine, CXC chemokine, and adhesion molecule mRNA and protein expression occurred in each model . In conclusion, i.t . or i.v . challenge with C . parvum in sensitized rats caused pulmonary granulomatous inflammation that was histologically similar to human extrinsic allergic alveolitis and sarcoidosis, respectively . Although the soluble and cellular mediators of granulomatous inflammation were qualitatively similar in both disease models, the differing anatomic source of the same antigenic challenge was responsible for differing clinicopathological presentations. Biochemistry, 1996 Sep 24, 35(38), 12292 - 302 High-resolution structure of the diphtheria toxin repressor complexed with cobalt and manganese reveals an SH3-like third domain and suggests a possible role of phosphate as co-corepressor; Qiu X et al.; The crystal structure of diphtheria toxin repressor (DtxR) in complex with the corepressor Co2+ has been determined at 2.0 A resolution and in complex with Mn2+ at 2.2 A resolution . The structure of the flexible third domain could be determined at this high resolution . It appears to contain five antiparallel strands exhibiting a fold very similar to the SH3 domain . A superposition of 46 equivalent C alpha atoms of DtxR and alpha-spectrin SH3 resulted in an rms deviation of 3.0 A . The sequence identity is only 7% . This third domain of DtxR appears to have no interactions with the DNA binding domain nor with the metal binding domain of the repressor . Yet, flexibility in the region between the second and the third domain allows in principle significant conformational changes such as might occur upon DNA binding . The two metal binding sites in the second domain have been unraveled in considerable detail . Metal binding site 1 was well occupied in both the cobalt and manganese structures and showed a surprising sulfate ion as ligand . The sulfate was proven beyond doubt by the high peak at its position in a selenate versus sulfate difference Fourier . The presence of the intriguing sulfate ion at such a crucial position near the metal corepressor suggests the possibility that under physiological conditions phosphate may act as a "co-corepressor" for this class of metal-regulated DNA binding proteins in Corynebacteria, Mycobacteria, and related organisms . The second metal binding site is significantly different in these two DtxR structures . In the 2.0 A cobalt structure, the site is not occupied by a metal ion . In the 2.2 A manganese structure the site is well occupied, at approximately the same position as observed previously in cadmium DtxR . The ligands are Glu105, His106, the carbonyl oxygen of Cys102, and a water molecule . The reasons for differential occupancy of this site in different structures are intriguing and require further investigations. Mol Gen Genet, 1996 Sep 13, 252(3), 255 - 65 A physical and genetic map of the Corynebacterium glutamicum ATCC 13032 chromosome; Bathe B et al.; A combined physical and genetic map of the Corynebacterium glutamicum ATCC 13032 chromosome was constructed using pulsed-field gel electrophoresis (PFGE) and hybridizations with cloned gene probes . Total genomic DNA was digested with the meganucleases SwaI (5'-ATTTAAAT-3'), PacI (5'-TTAATTAA-3'), and PmeI (5'-GTTTAAAC-3') yielding 26,27, and 23 fragments, respectively . The chromosomal restriction fragments were then separated by PFGE . By summing up the lengths of the fragments generated with each of the three enzymes, a genome size of 3082 +/- 20 kb was determined . To identify adjacent SwaI fragments, a genomic cosmid library of C.glutamicum was screened for chromosomal inserts containing SwaI sites . Southern blots of the PFGE gels were hybridized with these linking clones to connect the SwaI fragments in their natural order . By this method, about 90% of the genome could be ordered into three contigs . Two of the remaining gaps were closed by cross-hybridization of blotted SwaI digests using as probes PacI and PmeI fragments isolated from PFGE gels . The last gap in the chromosomal map was closed by hybridization experiments using partial SwaI digestions, thereby proving the circularity of the chromosome . By hybridization of gene probes to SwaI fragments separated by PFGE about 30 genes, including rRNA operons, IS element and transposon insertions were localized on the physical map. Zh Mikrobiol Epidemiol Immunobiol, 1996 Sep-Oct, (5), 73 - 5 {The genetic typing of Corynebacterium diphtheriae strains by the polymerase chain reaction with universal primers}; Mokrousov IV et al.; The use of polymerase chain reaction (UP-PCR with universal primers for the genetic typing of 85 C . diphtheriae strains, isolated from patients and carriers at the period of the epidemic, has shown that a single natural clone of biovar gravis seems to prevail in North Western Russia . UP-PCR is an effective too for the rapid intraspecific typing of strains on the molecular-genetic level and for the study of the circulation of C.diphtheriae epidemic clones. Vet Q, 1996 Sep, 18(3), 87 - 9 Mastitis in dairy cattle caused by Corynebacterium pseudotuberculosis and the feasibility of transmission by houseflies . I; Yeruham I et al.; Morbidity due to Corynebacterium pseudotuberculosis infection occurred in 29 dairy herds . The disease appeared basically in three clinical forms: cutaneous, mastitic, and visceral . The appearance of the disease showed a marked seasonality: in 23 herds it occurred during the spring and summer months (dry season) (March-October) . The mastitic form occurred in only 10 herds and the causative bacterium was isolated from 33 cows (5.8%) . All the strains of C . pseudotuberculosis isolated from the milk samples were found not to be nitrate reducers . The bacterium was excreted in the milk of six cows from herd B during a period of 11 months . In the mastitic cows, a decrease in milk production and considerable increases in the somatic cell count were noted . C . pseudotuberculosis was isolated from houseflies collected over a cow lesion . Laboratory-reared houseflies were successfully infected with C . pseudotuberculosis-contaminated milk, broth and sugar cubes . Flies infected with the bacterium from contaminated milk excreted the bacterium in their droppings for up to 4 h and from their saliva for up to 3 h post infection . The bacterium survived on the external organs of houseflies for no longer than 10 min post infection, after the flies had been dipped in contaminated broth. Clin Infect Dis, 1996 Sep, 23(3), 442 - 8 Microbiology and laboratory diagnosis of upper respiratory tract infections; Carroll K et al.; In the article that follows, Carroll and Reimer address a number of issues related to the clinical and laboratory diagnosis of upper respiratory tract infections . These syndromes occur with great frequency in both adults and children and have tremendous economic impact, related not only to lost productivity in the workplace but also to the frequent prescription by physicians of antibiotics, even when the etiologic agents of infection almost certainly are not bacteria . Most of these infections are diagnosed clinically, and specimens for microbiological identification are not obtained . Indeed, the difficulty in obtaining microbiological specimens that are not contaminated by resident colonizing flora often results in laboratory culture reports of dubious clinical value . As the authors note, the most standardized procedures are for the diagnosis of pharyngitis due to Streptococcus pyogenes . The preferred culture methods are reviewed as are the sensitivities, specificities, and limitations of rapid direct tests for group A streptococcal antigens . Currently, as the authors emphasize, a negative direct test mandates a conventional culture for S . pyogenes . More problematic are requests for isolation of other streptococci, Haemophilus species, corynebacteria, and gram-negative bacteria . Given limited resources, cost-containment imperatives, and the absence of clear evidence that these organisms are pharyngeal pathogens associated with important sequelae, my laboratory does not attempt to isolate these bacteria unless the ordering physician has directly consulted with me (the laboratory director) . Carroll and Reimer emphasize that nasopharyngeal cultures have no place in the microbiological diagnosis of otitis media and that diagnostic tympanocentesis is the only procedure for obtaining specimens that yield reliable microbiological findings . They also point out the futility of using swabs to obtain material for the diagnosis of otitis externa, since the external auditory canal cannot be decontaminated sufficiently to obtain a meaningful culture result . Finally, the authors address the available methods for obtaining specimens to establish the etiology of sinusitis . For microbiological diagnosis, direct antral puncture has been the method of choice for many years . However, otorhinolaryngologists now obtain many specimens endoscopically . It probably is not possible to obtain specimens by this method without contamination by normal upper respiratory flora . Thus, results of cultures of endoscopic specimens are more difficult to interpret . For patients with complicated illnesses, use of the diagnostic "gold standard" of antral puncture, as well as biopsy with histopathologic correlation, should be encouraged. J Clin Microbiol, 1996 Sep, 34(9), 2089 - 94 Patient-to-patient spread of a single strain of Corynebacterium striatum causing infections in a surgical intensive care unit; Brandenburg AH et al.; Over a 12-month period, Corynebacterium striatum strains were isolated from clinical specimens from 14 patients admitted to a surgical intensive care unit . These isolates were identical by morphology and biotype and displayed the same antibiogram . Ten isolates were found to be the sole possible pathogen . These 10 isolates were from six patients, three of whom had signs of infection at the time of positive culture . Further typing was performed by random amplification of polymorphic DNA analysis, by which all strains were identical and were found to differ to various degrees from reference strains and from isolates found in clinical samples from other wards . In a case-control study the only independent risk factor for acquiring the strain was intubation for longer than 24 h (odds ratio, 20.09; 95% confidence interval, 2.29 to 176.09) . The same strain was isolated from surfaces and from air sampled in the direct vicinity of infected patients but never from surfaces or air in other places of the ward . The strain was not isolated from the ventilators . The strain was cultured from the hands of personnel attending to infected patients, but no long-term carriers were found among members of the hospital personnel, suggesting transient carriage only . We conclude that C . striatum can cause serious nosocomial infections in surgical intensive care unit patients and may spread from patient to patient via the hands of attending personnel. Int J Lepr Other Mycobact Dis, 1996 Sep, 64(3), 274 - 81 Microbial colonizers in leprosy skin ulcers and intensity of inflammation; Sturm AW et al.; The microflora of 55 patients with leprosy skin ulcers was studied and related to a weighted inflammatory score (IS) . The control group consisted of 18 ulcers with different underlying pathology . Leprosy ulcers were characterized by the exclusive presence of two types of branching gram-positive rods; a particular interesting proposal is that Mycobacterium leprae share common antigens with these unusual "leprosy ulcer associated" organisms and group G beta-hemolytic streptococci . In the leprosy group, corynebacteria and branching rods accounted for 97% of gram-positive bacilli and Bacillus species constituted only 3% . In the control group, B . species formed 50% of gram-positive rods; the rest were corynebacteria (p = 0.03) . In the leprosy group, one third of the gram-positive bacteria were branching rods; none of them was acid fast . Ten of them were identified as Arcanobacterium haemolyticum, and the remaining 7 could not be identified . The IS of leprosy patients was lower than in the control group . The presence of more than two species of facultative or aerobic gram-negative rods or single species of pyogenic gram-positive cocci correlated with a high IS . The presence of two or more different pyogenic cocci resulted in a lower IS . Further studies into the nature of leprosyunique organisms as well as the inflammation inhibition factors in mixed infections are warranted . It is recommended that management of ulcers should consist of the application of local disinfection and early treatment of episodes of inflammation with a combination of fluoroquinolone and penicillin. Am J Clin Pathol, 1996 Sep, 106(3), 378 - 83 Heterogeneity within Corynebacterium minutissimum strains is explained by misidentified Corynebacterium amycolatum strains; Zinkernagel AS et al.; Forty-eight clinical strains that were tentatively identified as Corynebacterium minutissimum on the basis of standard biochemical reactions (Hollis-Weaver tables) as well as by the use of the API (RAPID) Coryne system were examined further . Two different groups of strains were observed . The first group (including the type strain of C minutissimum) contained 27 strains showing creamy colonies . These strains grew homogeneously in 6.5% NaCl broth, exhibited DNase activity, were susceptible to the vibriocidal compound O/129, produced succinic acid, and contained mycolic acids . The second group comprised 21 strains with dry colonies . They grew in clumps at the surface of 6.5% NaCl broth, DNase activity was not detected, they were resistant against O/129, produced large amounts of propionic acid, and mycolic acids were not detected . In combination with quantitative DNA-DNA hybridizations, it was demonstrated that strains of the second cluster belonged, in fact, to C amycolatum . Furthermore, it was observed that a few C minutissimum strains may also ferment mannitol . These data indicate that the clinical microbiologist must be careful not to misidentify C amycolatum strains as C minutissimum. FEMS Microbiol Lett, 1996 Sep 1, 142(2-3), 139 - 45 Cloning and manipulation of the Corynebacterium pseudotuberculosis recA gene for live vaccine vector development; Pogson CA et al.; Corynebacterium pseudotuberculosis is an intracellular bacterial pathogen causing a chronic abscessing disease in sheep and goats called caseous lymphadenitis . We are developing this bacterial species as a live vector system to deliver vaccine antigens to the animal immune system . Foreign genes expressed in bacterial hosts can be unstable so we undertook to delete the C . pseudotuberculosis chromosomal recA gene to determine whether a recA- background would reduce the frequency of recombination in cloned DNA . Homologous DNA recombination within an isogenic recA- C . pseudotuberculosis was 10-12-fold lower than that in the recA+ parental strain . Importantly, the recA mutation had no detectable affect upon the virulence of C . pseudotuberculosis in a mouse model . Taken together these results suggest that a recA- background may be useful in the further development of C . pseudotuberculosis as a vaccine vector. Appl Environ Microbiol, 1996 Sep, 62(9), 3171 - 5 Purification and partial characterization of a novel antibacterial agent (Bac1829) Produced by Staphylococcus aureus KSI1829; Crupper SS et al.; A novel antimicrobial agent from Staphylococcus aureus KSI1829, designated Bac1829, was purified by sequential steps of ammonium sulfate precipitation, Sephadex G-50 gel filtration chromatography, and hydrophobic interaction chromatography . Purified Bac1829 has a molecular mass of 6,418 +/- 2 Da . The peptide in heat stable, since full biological activity is retained after heating at 95 degrees C for 15 min, and it is destroyed by digestion with proteases . Amino acid sequence analysis revealed a high concentration of Ala and Gly residues, which respectively comprised 24 and 19% of the total amino acid content . Additionally, high levels of hydrophobic amino acids were present, accounting for the hydrophobic nature of Bac1829 . Purified Bac1829 killed exponentially growing Corynebacterium renale in a dose-dependent manner by a bactericidal mode of action . A partial inhibitory spectrum analysis revealed that the following organisms were sensitive to the inhibitory activity of Bac1829: S . aureus RN4220, Streptococcus suis, Corynebacterium pseudotuberculosis, C . renale, Corynebacterium diptheriae, Haemophilus parasuis, Bordetella pertussis, Bordetella bronchoseptica, Moraxella bovis, and Pasteurella multocida. J Bacteriol, 1996 Sep, 178(17), 5229 - 34 Isolation, characterization, and expression of the Corynebacterium glutamicum betP gene, encoding the transport system for the compatible solute glycine betaine; Peter H et al.; Corynebacterium glutamicum accumulates glycine betaine under conditions of high osmolarity . Previous work revealed the existence of a high-affinity glycine betaine permease which is osmotically regulated . In the present study, the corresponding gene was cloned . The betP gene, encoding the glycine betaine uptake carrier, was isolated by heterologous complementation of mutant strain Escherichia coli MKH13 . From sequence analysis it is predicted to encode a protein of 595 amino acids . This protein shares 36% identity with the choline transport system BetT and 28% identity with the carnitine transport system CaiT of E . coli, as well as 38% identity with a protein with an unknown function from Haemophilus influenzae . Analysis of hydropathy indicated a common structure for all four transport proteins . After heterologous expression of betP in E . coli MKH13, the measured Km values for glycine betaine and the cotransported Na+ were similar to those found in C . glutamicum, whereas the modulation of activity by osmotic gradients was shifted to lower osmotic values. J Urol, 1996 Sep, 156(3), 881 - 4 Insertion of a double pigtail ureteral stent for the prevention of urological complications in renal transplantation: a prospective randomized study; Benoit G et al.; PURPOSE: Urologists successfully use ureteral stents to protect the ureterovesical anastomosis in nontransplant patients . MATERIALS AND METHODS: We determined the value of ureteral stents in transplant patients . The frequency of urological complications (leaks, obstructions and urinary tract infections) was compared in a prospective randomized series of 194 kidney transplantations (97 with and 97 without a double pigtail ureteral stent) . RESULTS: In the stent group 1 patient had a urinary leak and 35 had urinary tract infections (including 2 cases of Corynebacterium cystitis) . In the no stent group 6 patients had urinary leaks, 4 had obstructions and 32 had urinary tract infections . The 1-year patient and graft survival rates were similar in both groups, and renal function at 1 year was also similar (229 versus 208 mumol./l . creatinine in the stent and no stent groups, respectively) . A small number of stent related complications occurred (2 stent breakages and 1 stent migration) . No stones formed in any case . CONCLUSIONS: Ureteral stent insertion significantly decreases the rate of vesicoureteral leakage and obstruction in renal transplantation. Proc Natl Acad Sci U S A, 1996 Aug 20, 93(17), 9138 - 41 Iron chelates bind nitric oxide and decrease mortality in an experimental model of septic shock; Kazmierski WM et al.; The hydroxamic acid siderophore ferrioxamine B {FeIII(HDFB)+} and the iron complex of diethylenetri-aminepentaacetic acid {FeIII(DTPA)2-} protected mice against death by septic shock induced by Corynebacterium parvum + lipopolysaccharide . Although FeIII(DTPA)2- was somewhat more effective than FeIII(HDFB)+, the iron-free ligand H4DFB+ was significantly more effective than DTPA . The hydroxamic acid chelator has a much higher iron affinity than the amine carboxylate, allowing for more efficient formation of the FeIII(HDFB)+ complex upon administration of the iron-free ligand . Electrochemical studies show that FeIII(DTPA)2- binds NO stoichiometrically upon reduction to iron(II) at biologically relevant potentials to form a stable NO adduct . In contrast, FeIII(HDFB)+ is a stable and efficient electrocatalyst for the reduction of NO to N2O at biologically relevant potentials . These results suggest that the mechanism of protection against death by septic shock involves NO scavenging and that particularly effective drugs that operate a low dosages may be designed based on the principle of redox catalysis . These complexes constitute a new family of drugs that rely on the special ability of transition metals to activate small molecules . In addition, the wealth of information available on siderophore chemistry and biology provides an intellectual platform for further development. J Am Vet Med Assoc, 1996 Aug 15, 209(4), 804 - 9 Corynebacterium pseudotuberculosis infection in horses: 538 cases (1982-1993); Aleman M et al.; OBJECTIVE--To describe clinical manifestations of Corynebacterium pseudotuberculosis infection in horses and to evaluate diagnostic methods for identification of this disease . DESIGN--Retrospective case series . ANIMALS--538 horses with a diagnosis of C pseudotuberculosis infection . RESULTS--Median age of horses with external abscesses was similar to that in horses with internal abscesses . Breed and sex did not appear to be associated with infection . Cases were detected during all 12 months; however, the disease was most common in the fall and early winter, with the highest incidence in September, October, and November in every year . Most horses (492/538, 91.4%) had a single episode of infection, without recurrence in subsequent years . Of 538 horses, 308 had pectoral abscesses, although infection was documented in many other anatomic locations . Forty-two horses had internal abscesses involving the abdomen or thoracic cavity . Corynebacterium pseudotuberculosis infection was readily identified by bacterial culture of aspirate samples from abscesses . The synergistic hemolysis inhibition test was useful for diagnosis of internal abscesses; however, it was unreliable for the diagnosis of external abscesses . Horses with external abscesses responded well to conventional treatment, in contrast to those with internal abscesses . The overall case fatality was low (3.9%), and was considerably lower for horses with external abscesses (0.8%) than for horses with internal abscesses (40.5%) . CLINICAL IMPLICATIONS--Serology (synergistic hemolysis inhibition titers > or = 512) is useful for diagnosis of internal abscesses, but not reliable for diagnosis in horses with external abscesses . Prognosis for horses with internal abscesses is considerably poorer than for those with external abscesses.
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