J Clin Microbiol, 1994 Nov, 32(11), 2801 - 12 The superoxide dismutase gene, a target for detection and identification of mycobacteria by PCR; Zolg JW et al.; The superoxide dismutase gene has been identified as a target in screening for the presence of mycobacteria on the genus level and differentiating relevant mycobacterial species from one another by PCR . Consensus primers deduced from known superoxide dismutase gene sequences allowed the amplification of DNAs from a variety of bacteria, fungi, and protozoa . Selected amplicons from Actinomyces viscosus, Corynebacterium diphtheriae, Corynebacterium pseudodiphtheriticum, Mycobacterium avium, M . fortuitum, M . gordonae, M . intracellulare, M . kansasii, M . scrofulaceum, M . simiae, M . tuberculosis, M . xenopi, and Nocardia asteroides were subsequently cloned and sequenced . The alignment of those sequences facilitated the selection of primers targeting conserved regions present in myobacterial species but absent in nonmycobacterial species and thus allowed the genus-specific amplification of all 28 different mycobacterial species tested . A pool of genus-specific allowed the genus-specific amplification of all 28 different mycobacterial species tested . A pool of genus-specific probes recognized 23 of the 28 mycobacterial species and did not cross-react with any of the 96 nonmycobacterial species tested . In addition, probes recognizing species-specific variable regions within the superoxide dismutase genes of M . avium, M . fortuitum, M . gordonae, M . intracellulare, M . kansasii, M . scrofulaceum, M . simiae, the M . tuberculosis complex, and M . xenopi were identified . All probes recognized only the species from which they were derived and did not cross-react with any other mycobacterial species or with any of the nonmycobacterial species tested . We conclude that the superoxide dismutase gene is a suitable target for amplifying mycobacteria by PCR on the genus level, confirming correct amplification by genus-specific probes, and differentiating relevant species from one another by a set of species-specific probes. J Clin Microbiol, 1994 Nov, 32(11), 2686 - 91 Primary identification of Aureobacterium spp . isolated from clinical specimens as "Corynebacterium aquaticum"; Funke G et al.; Over a 6-year period 11 yellow-pigmented gram-positive rods (GPRs) with an oxidative carbohydrate metabolism were isolated from clinical specimens or were received as reference cultures and tentatively identified as "Corynebacterium aquaticum" according to the guide of Hollis and Weaver for the differentiation of GPRs (D . G . Hollis and R . E . Weaver, Gram-Positive Organisms: a Guide to Identification, 1981) . Because these isolates seemed to be rather heterogeneous, comparative analyses with the type strain of "C . aquaticum" as well as six type strains of species belonging to the genus Aureobacterium were performed by biochemical and chemotaxonomic methods . Only four clinical strains were found to be "C . aquaticum," whereas seven strains were found to belong to the genus Aureobacterium . Discriminative phenotypic reactions between "C . aquaticum" and Aureobacterium spp . included hydrolysis of gelatin and casein (both reactions negative for "C . aquaticum" strains but positive for most Aureobacterium strains) . Moreover, peptidoglycan analysis provided a reliable means of differentiating yellow-pigmented GPRs at the genus level (diaminobutyric acid as the interpeptide bridge in "C . aquaticum" and glycine-ornithine as the interpeptide bridge in Aureobacterium spp.) . Antimicrobial susceptibility testing revealed that vancomycin showed an intermediate MIC for three of the four clinical "C . aquaticum" isolates, whereas all Aureobacterium strains were susceptible to vancomycin . To our knowledge, this is the first report outlining the isolation of Aureobacterium spp . from clinical specimens . However, Aureobacterium isolates could not be identified to the species level by the tests used in the study. J Dairy Sci, 1994 Nov, 77(11), 3331 - 7 Efficacy of dry cow therapy and a Propionibacterium acnes product in herds with low somatic cell count; Hogan JS et al.; Dry cow therapy and a Propionibaterium acnes product were evaluated in four commercial herds with low SCC . Cows were randomly assigned within herds to treatment groups of approximately 90 cows receiving dry cow therapy, P . acnes, dry cow therapy plus P . acnes, or no treatment in a factorial arrangement . Each lactating quarter of cows that received dry cow therapy was infused via the teat duct with 300 mg of cephaprin at drying off . Cows that received P . acnes were infused intravenously with .4 mg of killed P . acnes at drying off, 7 to 10 d prepartum, and within 7 d after calving . A second prepartum injection of P . acnes immunostimulator was administered to cows that did not calve within 10 d after the first prepartum injection . Dry cow therapy enhanced bacteriological cures of IMI by Staphylococcus aureus and Corynebacterium bovis at drying off . Dry cow therapy reduced incidence of new IMI by environmental streptococci and C . bovis that originated during the dry period . Cows treated with P . acnes alone had a greater incidence of new IMI by Gram-negative bacilli originating during the dry period than did cows in the other treatment groups . Incidence of clinical mastitis at calving was greater for cows receiving no treatment than for cows receiving dry cow therapy, P . acnes, or dry cow therapy plus P . acnes. Microbiology, 1994 Nov, 140 ( Pt 11), 3099 - 108 Malate synthase from Corynebacterium glutamicum: sequence analysis of the gene and biochemical characterization of the enzyme; Reinscheid DJ et al.; Malate synthase is one of the key enzymes of the glyoxylate cycle and is essential for growth on acetate as sole carbon source . The aceB gene from Corynebacterium glutamicum, encoding malate synthase, was isolated, subcloned and expressed in Escherichia coli and C . glutamicum . Sequencing of a 3024 bp DNA fragment containing the aceB gene revealed that it is located close to the isocitrate lyase gene aceA . The two genes are separated by 597 bp and are transcribed in divergent directions . The predicted aceB gene product consists of 739 amino acids with an M(r) of 82,362 . Interestingly, this polypeptide shows only weak identity with malate synthase polypeptides from other organisms and possesses an extra N-terminal sequence of about 170 amino acid residues . Inactivation of the chromosomal aceB gene led to the absence of malate synthase activity and to the inability to grow on acetate, suggesting that only one malate synthase is present in C . glutamicum . The malate synthase was purified from an aceB-overexpressing C . glutamicum strain and biochemically characterized . The native enzyme was shown to be a monomer migrating at an M(r) of about 80,000 . By sequencing the N-terminus of malate synthase the predicted translational start site of the enzyme was confirmed . The enzyme displayed Km values of 30 microM and 12 microM for the substrates glyoxylate and acetyl CoA, respectively . Oxalate, glycolate and ATP were found to be inhibitors of malate synthase activity . The present study provides evidence that the malate synthase from C . glutamicum is functionally similar to other malate synthase enzymes but is different both in size and primary structure. Mol Gen Mikrobiol Virusol, 1994 Nov-Dec, (6), 27 - 9 {Use of a DNA-DNA hybridization method for studying Corynebacterium diphtheriae strains}; Mikhailovich VM et al.; In addition to traditional tests, a battery of DNA probes was used to characterize 134 toxigenic and 125 nontoxigenic C.diphtheriae strains isolated in Russia in 1986-1989 . 2.5% of nontoxigenic strains carried the determinants of both toxin subunits A and B, and 20% possessed at least a fragment of the gene determining toxin synthesis . Optimal conditions of hybridization technology were found . The method may be used as an alternative to traditional techniques in diagnostic studies and screenings. Zentralbl Bakteriol, 1994 Nov, 281(4), 549 - 55 The influence of Propionibacterium acnes (Corynebacterium parvum) fractions on immune response in vivo; Mara M et al.; Bacterin of Propionibacterium acnes (Corynebacterium parvum), its cellular fractions (lipids, fractions obtained by mechanical disruption and differential centrifugation, by phenol-water and pyridine extractions), and a polysaccharide from culture filtrate were prepared and tested in mice . The activation of RES by splenomegaly and hepatomegaly, prevention of listerial infection, prevention of the lethal effect of sarcoma 180, and depression of liver microsomal cytochrome P-450 were employed . The bacterin was effective in all tests . Lipid-free cells were less active, in particular in the activation of RES and in the listerial infection model . Fractions prepared by the disruption and differential centrifugation lost their activity in all tests along with a decrease in molecular weight . Lipids extracted by ethanol caused pronounced splenomegaly and decreased the cytochrome P-450 content . The residue left after the phenol-water extraction was very active, its delipidation did not destroy the activity . Pyridine extraction provided a completely inactive extract, but a very active residue . The possibility of reducing the complexity of bacterin while preserving immunomodulatory effect is demonstrated. Minerva Chir, 1994 Nov, 49(11), 1077 - 82 {Treatment of malignant pleural effusion by percutaneous catheter drainage and chemical pleurodesis}; Chella A et al.; From June 1989 to December 1991, 93 patients affected by malignant pleural effusion (MPE) were treated as out patients using a ultrasound (US) guided catheter drainage (10F Percuflex pig-tail catheter-Medi-tech@(PTC) and chemical pleurodesis . The PTC was positioned when the MPE had been diagnosed by X-ray and all the patients had undergone some thoracentesis (mean 3.2, range 2-6) . We started a chemical pleurodesis when the MPE was inferior to 100cc/day injecting Epidoxorubicin (EDX 30 mg/mq) in the pleural space on alternate day for three administrations, and then, a lyophilized Corynebacterium Parvum (Wellcome strain CN 6134) (CBP 14 mg) on alternate day for two administrations . The complete response rate of this treatment was 90% (86 pt) with a mean treatment period of 32.9 days (range 12-62) . Complete response was assessed as total resolution of pleural effusion . Side effects were short-term hyperpyrexia in 65 cases (69.8%) and pleurodynia in 15 cases (15.7%) . Three patients (3.1%) were complicated by a pleural infection which was resolved in 1 case and became chronic in 2 . These findings indicate that this technique is an adequate treatment for the control of MPE inducing a clinical and quality-life improvement. FEMS Microbiol Lett, 1994 Nov 1, 123(3), 261 - 7 A phylogeny of the genus Nocardia deduced from the analysis of small-subunit ribosomal DNA sequences, including transfer of Nocardia amarae to the genus Gordona as Gordona amarae comb . nov; Ruimy R et al.; According to phylogenetic analyses of nearly complete small-subunit ribosomal DNA sequences, the genus Nocardia should not comprise the two species Nocardia petroleophila and Nocardia amarae . N . amarae should be reassigned to the genus Gordona as Gordona amarae . All of the other Nocardia species form a monophyletic unit, closely related to species of the genus Rhodococcus . It is proposed to revive the name 'CMN' to comprise the genera Corynebacterium, Tsukamurella, Mycobacterium, Gordona, Rhodococcus and Nocardia that form a well identified and monophyletic unit . They are all characterized by a cell wall chemotype IV with mycolic acids. Biochem Biophys Res Commun, 1994 Oct 28, 204(2), 635 - 40 Inhibition of nitric oxide synthesis enhances the expression of inducible nitric oxide synthase mRNA and protein in a model of chronic liver inflammation; Luss H et al.; In septic shock the inhibition of inducible nitric oxide synthase (iNOS) could be of therapeutic value . However, side effects have to be investigated . Therefore we studied the effects of chronic NOS inhibition on the level of iNOS expression in a model of chronic liver inflammation induced by Corynebacterium parvum (C . parvum) which causes sustained iNOS expression in the liver . NOS inhibitors decreased the rise in plasma levels and urinary excretion of nitrite/nitrate by about 50%; however, iNOS mRNA and protein were increased to 200% and 150%, respectively . Thus chronic inhibition of NOS can result in an increase in iNOS mRNA level and protein under conditions when iNOS is expressed . This could result in an overproduction of NO upon removal of the NOS-inhibitor. Cancer Res, 1994 Oct 15, 54(20), 5420 - 3 B7-1/CD80-transduced tumor cells elicit better systemic immunity than wild-type tumor cells admixed with Corynebacterium parvum; Chen L et al.; Tumor cells genetically modified by transduction of B7 (B7-1/CD80), a natural ligand for the T-cell costimulatory molecules CD28 and CTLA-4, can elicit potent tumor immunity, and they can be effective for treatment of established cancers in animal models . In this study, three tumor lines, the EL4 lymphoma, the P815 mastocytoma, and the MCA102 sarcoma were transduced with recombinant retrovirus containing the murine B7 gene, and their potency to induce systemic immunity protective against challenge with wild-type tumor was compared to that of the same tumor cells admixed with the commonly used adjuvant Corynebacterium parvum . While admixture of tumor cells with C . parvum resulted in complete regression of tumors in syngeneic mice, it did not induce protective immunity against a subsequent challenge of wild-type cells from any of the 3 tumors tested . In contrast, B7-transduced EL4 and P815 tumors regressed locally and induced a potent systemic immunity to wild-type tumors and a higher level of cytotoxic T-cell activity than did tumor cells admixed with C . parvum . No systemic immunity was induced by B7-transduced nonimmunogenic MCA102 sarcoma cells . Our results demonstrate that immunogenic tumor cells transduced with the B7 gene are superior to tumor cells mixed with C . parvum for the induction of systemic tumor immunity. Biochem J, 1994 Oct 1, 303 ( Pt 1), 323 - 7 Phosphinic peptide analogues as potent inhibitors of Corynebacterium rathayii bacterial collagenase; Yiotakis A et al.; Pseudo-substrate analogues of collagenase from Corynebacterium rathayii, in which the scissile peptide bond is replaced by a phosphinic moiety, were synthesized and evaluated as inhibitors of this enzyme . The phosphinic tetrapeptide, Z-Phe-psi(PO2CH2)-Gly-Pro-Nle (1), was found to be a potent inhibitor of collagenase with a Ki value of 8 nM . Increasing the length of the phosphinic-containing inhibitors from tetra- to hepta-peptide size further improves the potency of these compounds . The heptapeptide analogue, Z-Phe-Gly-Pro-Phe-psi(PO2CH2)-Gly-Pro-Nle-OMe, with a Ki value of 0.6 nM, is the most potent inhibitor reported to date for bacterial collagenases . A comparison between the phosphinic analogue Z-Phe-psi(PO2CH2)-Gly-Pro-Nle (1) and the phosphonamide peptide Z-Phe-psi(PO2NH)-Gly-Pro-Nle (2) shows that for bacterial collagenase the replacement of a CH2 by an NH group results only in a modest increase in affinity from Ki = 8 nM for compound 1 to Ki = 6 nM for compound 2 . Most of the phosphorus-containing inhibitors of this series are slow- or slow-tight-binding inhibitors with second-order rate constants for association and dissociation varying respectively for the kon values from 1 x 10(3) to 26 x 10(3) M-1.s-1 and for the koff values from 3 x 10(-4) to 2 x 10(-5) s-1 . Interestingly, the lower affinity of the molecule containing a D residue in the P1 position of the inhibitor, compared with the molecule with an L residue in this position, is mainly the consequence of a lower rate constant for association of these D stereoisomers with the enzyme . This study demonstrates that phosphinic peptide analogues are potent inhibitors of a bacterial collagenase . The development of new phosphinic peptides should lead to the discovery of potent inhibitors of other zinc metalloproteases . Details of how the analogues were synthesized are given in Supplementary Publication SUP 50176 (14 pages), which has been deposited with the British Library Document Supply Centre, Boston Spa, Wetherby, W . Yorkshire LS23 7BQ, from whom copies can be obtained on the terms indicated in Biochem . J . (1994) 297, 9. Hepatology, 1994 Oct, 20(4 Pt 1), 1055 - 60 Tumor necrosis factor-alpha regulates in vivo nitric oxide synthesis and induces liver injury during endotoxemia; Harbrecht BG et al.; Tumor necrosis factor-alpha is a principal mediator of the pathophysiological effects of endotoxemia and endotoxin shock . Tumor necrosis factor-alpha also contributes to the stimulation of nitric oxide synthesis by the induction of the enzyme nitric oxide synthase in a variety of tissues . Although the importance of tumor necrosis factor-alpha in the induction of nitric oxide synthase activity in vitro is well known, its role in in vivo nitric oxide synthesis has not been convincingly established . We were interested in determining whether tumor necrosis factor-alpha plays a significant role in the in vivo induction of nitric oxide synthesis . In Corynebacterium parvum-primed mice, lipopolysaccharide injection resulted in elevated serum tumor necrosis factor-alpha levels early and increased hepatic enzyme release (641 +/- 80 IU AST/L; 22.7 +/- 1.9 IU ornithine carbamoyltransferase per liter) and plasma nitrite and nitrate (804 +/- 84 mumol/L) 5 hr after lipopolysaccharide injection . Polyclonal rabbit anti-mouse anti-tumor necrosis factor-alpha reduced in vivo tumor necrosis factor-alpha levels (1 hr, 7,332 +/- 1,492 U tumor necrosis factor-alpha per milliliter) and reduced nitric oxide synthesis as measured by plasma nitrite and nitrate (352 +/- 69 mumol/L) . Polyclonal rabbit anti-mouse anti-tumor necrosis factor-alpha also reduced lipopolysaccharide-induced hepatic enzyme release (428 +/- 33 IU AST/L; 16.0 +/- 2.5 IU ornithine carbamoyltransferase per liter) . NG-monomethyl-L-arginine, a competitive inhibitor of nitric oxide synthesis, also decreased plasma nitrite and nitrate (104 +/- 9 mumol/L) but increased the lipopolysaccharide-induced hepatic injury (797 +/- 66 IU AST/L; 33.1 +/- 2.1 IU ornithine carbamoyltransferase per liter).(ABSTRACT TRUNCATED AT 250 WORDS) Mol Microbiol, 1994 Oct, 14(2), 191 - 7 Iron, DtxR, and the regulation of diphtheria toxin expression; Tao X et al.; In recent years considerable advances have been made in the understanding of the molecular basis of iron-mediated regulation of diphtheria toxin expression . The tox gene has been shown to be regulated by the heavy metal ion-activated regulatory element DtxR . In the presence of divalent heavy metal ions, DtxR becomes activated and binds to a 9 bp interrupted palindromic sequence . The consensus-binding site has been determined by both the sequence analysis of DtxR-responsive operators cloned from genomic libraries of Corynebacterium diphtheriae as well as by in vitro genetic methods using cyclic amplification of selected targets (CASTing) . It is now clear that DtxR functions as a global iron-sensitive regulatory element in the control of gene expression in C . diphtheriae . In addition, the metal ion-activation domain of DtxR is being characterized by both mutational analysis and determination of the X-ray structure at 3.0 A resolution. Clin Infect Dis, 1994 Oct, 19(4), 746 - 50 Pyogenic and tuberculous spondylodiskitis (vertebral osteomyelitis) in 80 adult patients; Perronne C et al.; Bacterial spondylodiskitis--i.e., adjacent vertebral osteomyelitis and diskitis--was studied in 80 adult patients . The infection was due to Mycobacterium tuberculosis in 31 cases (39%) and to pyogenic bacteria in 49 cases (61%) . The latter pathogens included gram-negative bacilli in 16 cases (20%), Staphylococcus species in 15 (19%), Streptococcus species in 9 (11%), and Corynebacterium species in 1 (1%); the pathogens in the 8 remaining cases (10%) were not identified . Of the patients with tuberculous spondylodiskitis, 55% came from countries where tuberculosis is endemic (P < .001) . Cases due to staphylococci and those due to M . tuberculosis were associated with a high frequency of previous active infection with those respective organisms at any site (47% and 42%, respectively; P < .001) and with a high rate of neurological complications (33% and 32%, respectively; P < .001) . Nine patients with pyogenic spondylodiskitis (18%) but only one patient with tuberculous spondylodiskitis (3%) had diabetes mellitus (P < .05) . Blood cultures were positive in 23 (56%) of the 41 cases of pyogenic spondylodiskitis due to an identified bacterium . Discovertebral needle biopsy contributed to the bacteriologic diagnosis in 29 (74%) of 39 cases. Proc Natl Acad Sci U S A, 1994 Sep 27, 91(20), 9646 - 50 Determination of the minimal essential nucleotide sequence for diphtheria tox repressor binding by in vitro affinity selection; Tao X et al.; The expression of diphtheria toxin in lysogenic toxigenic strains of Corynebacterium diphtheriae is controlled by the heavy metal ion-activated regulatory protein DtxR . In the presence of divalent heavy metal ions, DtxR specifically binds to the diphtheria tox operator and protects a 27-bp interrupted palindromic sequence from DNase I digestion . To determine the consensus DNA sequence for DtxR binding, we have used gel electrophoresis mobility-shift assay and polymerase chain reaction (PCR) amplification for in vitro affinity selection of DNA binding sequences from a universe of 6.9 x 10(10) variants . After 10 rounds of in vitro affinity selection, each round coupled with 30 cycles of PCR amplification, we isolated and characterized a family of DNA sequences that function as DtxR-responsive genetic elements both in vitro and in vivo . Moreover, these DNA sequences were found to bind activated DtxR with an affinity similar to that of the wild-type tox operator . The DNA sequence analysis of 21 unique in vitro affinity-selected binding sites has revealed the minimal essential nucleotide sequence for DtxR binding to be a 9-bp palindrome separated by a single base pair. Biochem Biophys Res Commun, 1994 Sep 15, 203(2), 820 - 7 Direct evidence for a constitutive internal promoter in the tryptophan operon of Corynebacterium glutamicum; O'Gara JP et al.; Insertional inactivation of the trpE gene in Corynebacterium glutamicum AS019 blocked expression of the trp operon from the primary promoter and resulted in tryptophan auxotrophy . The presence of indole, the substrate for the terminal trp pathway enzyme tryptophan synthase, reversed this auxotrophy thereby providing evidence for the existence of an internal promoter independently expressing the tryptophan synthase genes (trpB and trpA) . The toxic tryptophan analogue 6-fluorotryptophan failed to inhibit the growth permitted by indole thus suggesting that expression of tryptophan synthase from the internal promoter is constitutive. FEMS Microbiol Lett, 1994 Sep 15, 122(1-2), 1 - 6 Bacterial oxidation of propane; Ashraf W et al.; The bacterial metabolism of propane and the pathway(s) involved are poorly understood, as the relative importance of terminal versus subterminal oxidation of propane, via propan-1-ol and propan-2-ol, respectively, is still unclear . In the case of bacteria, the ability to oxidize propane appears to be confined mainly to the Gram-positive Corynebacterium - Nocardia - Mycobacterium - Rhodococcus complex . Studies on propane oxidation have been hampered by a lack of firm enzymological data; for example, to date there are no reports of a purified propane oxygenase system . However, oxygenase activity has been confirmed by the production of propan-1-ol and/or propan-2-ol, and more recently by the co-oxidation of propene to 1,2-epoxypropane in cell extracts of propane-grown cells . Here, we review the use of genetic, biochemical and immunological techniques to assess the role(s) of terminal and subterminal oxidation in the metabolism of propane by Rhodococcus rhodochrous PNKb1 and present a general overview of the topic. J Urol, 1994 Sep, 152(3), 1002 - 4 Spontaneous urinary calculus in young LEW rats caused by Corynebacterium renale; Osanai T et al.; Spontaneous urinary calculus was observed in only 27% of female LEW rats among nine inbred strains maintained in the Institute for Animal Experimentation . The condition occurred between 3 and 9 weeks of age with loss of weight, enlargement of the os urethral externum, anuria and general marasmus . All of the affected rats died from anuria 3 to 4 days after the onset . The affected rats had one to ten infected stones or many fine crystals of sand, which consisted of magnesium ammonium phosphate, in the urinary bladder and/or urethra . Analysis of microorganisms revealed that Corynebacterium renale, known as a causative agent of bovine pyelonephritis, was the most probable candidate for producing infection stones . This is the first report that the rat is spontaneously infected with C . renale . This could be a useful animal model for human urinary calculus and also for C . renale infection in the field of veterinary science. Mol Microbiol, 1994 Sep, 13(5), 833 - 42 Threonine dehydratases of Corynebacterium glutamicum with altered allosteric control: their generation and biochemical and structural analysis; Mockel B et al.; Threonine dehydratase is the key enzyme in L-isoleucine synthesis, since it is allosterically feedback-inhibited by L-isoleucine . With the aim of obtaining regulatorily altered mutants of the threonine dehydratase of Corynebacterium glutamicum, amino acids were specifically exchanged and a new biological system of mutant selection was developed, based on the intoxication of Escherichia coli by ketobutyrate, which is the dehydratase reaction product . A collection of 19 mutant enzymes was generated and genetically and biochemically characterized comprising a whole range of regulatorily and catalytically altered enzymes . Of particular interest is the mutant Val-323-Ala, which is characterized by the fact that the L-isoleucine inhibition is entirely abolished so that the enzyme is always present in a relaxed, high-activity state . Correspondingly, the Hill coefficient is 1.4, in contrast to the value of 3.4 characteristic of the wild-type enzyme . Another peculiar mutant generated is the double mutant His-278-Arg-Leu-351-Ser . Here, again, L-isoleucine no longer inhibits catalytic activity, but the effector still promotes major structural changes of the protein, as ascertained from the L-isoleucine-dependent loss of pyridoxal-5'-phosphate from this mutant enzyme . Further enzymes obtained are reduced in L-isoleucine inhibition to a varying degree . Detailed studies on the structure of the enzyme revealed a partially very high similarity of the secondary structure to the mechanistically identical beta-subunit of the tryptophan synthase . This provides further indications concerning the localization of the regulatory and catalytic domain of the threonine dehydratase. J Bacteriol, 1994 Sep, 176(18), 5718 - 28 Molecular analysis and characterization of a broad-host-range plasmid, pEP2; Zhang Y et al.; Plasmid pEP2 was found to encode a protein, RepA, which is essential and rate limiting for its replication in Escherichia coli and Corynebacterium pseudotuberculosis . Mutations which altered the rate of synthesis of this protein in E . coli affected the copy number and segregational stability of pEP2 in the two hosts . RepA contains 483 amino acid residues and has the calculated molecular weight of 53,925 . It shows 45% amino acid residue identity with open reading frame ORF2 of pSR1, a plasmid isolated from Corynebacterium glutamicum (J . A . C . Archer and A . J . Sinskey, J . Gen . Microbiol . 139:1753-1759, 1993) . Plasmid pEP2 was shown to accumulate single-stranded DNA corresponding to the RepA coding strand during its replication in E . coli and C . pseudotuberculosis, suggesting that it may replicate by a rolling circle mechanism . However, RepA has no significant sequence homology with the replication initiator proteins of plasmids known to use this mode of replication. J Med Chem, 1994 Aug 19, 37(17), 2713 - 20 Cyclic peptides with a phosphinic bond as potent inhibitors of a zinc bacterial collagenase; Yiotakis A et al.; A series of cyclic peptides containing a phosphinic bond were synthesized and evaluated as inhibitors of a zinc bacterial collagenase from Corynebacterium rathaii . Among this series of pseudopeptides of different sizes of cycles, only two molecules Ia (cyclo{Gly-Pro-Phe psi(PO2CH2)-Gly-Pro-Ahx}) and Va (cyclo{beta Ala-Pro-Phe psi (PO2CH2)Gly-Pro-Ahx}) were found to be rather potent inhibitors of this protease, with Ki values of 120 and 90 nM, respectively . Besides the influence of the peptide ring size, this study suggests that both the stereochemical and the conformational properties of the pseudophenylalanine residue in these cyclic peptides may determine their potency . Interestingly, the kinetic analysis for the binding of the cyclic peptide inhibitors Ia and Va to the collagenase, as compared to a linear parent compound, reveals that the lower potency of the cyclic peptides is mostly the consequence of a lower rate constant for association to the enzyme . To our knowledge, this is the first report on cyclic phosphinic peptides and on their activities as inhibitors of a zinc protease. J Clin Microbiol, 1994 Aug, 32(8), 1860 - 5 Genomic diversity among Corynebacterium jeikeium strains and comparison with biochemical characteristics and antimicrobial susceptibilities; Riegel P et al.; Levels of DNA relatedness were determined by performing DNA-DNA hybridization experiments (S1 nuclease procedure) with 13 human isolates exhibiting various antimicrobial susceptibility patterns which had been identified as Corynebacterium jeikeium by classical tests and the API Coryne system and with reference strains of C . jeikeium and related taxa . Twelve of 13 isolates which formed three genomic groups showed between 22 and 75% relatedness with the type strain of C . jeikeium . One of these genomic groups included all the strains resistant to penicillin and gentamicin and is genomically related to the C . jeikeium type strain at the species level . In addition, the reference strain of "Corynebacterium genitalium" biotype II was found to belong to this genospecies and therefore can be considered as a synonym of C . jeikeium . In contrast, one isolate and the reference strains of "Corynebacterium pseudogenitalium" biotypes C-3 and C-4 which were assigned to C . jeikeium by the API Coryne system were less than 10% related to the C . jeikeium type strain . These nongenomically related strains can be differentiated from the jeikeium-related strains on the basis of positive acidification from fructose and growth under anaerobic conditions . Furthermore, these strains exhibited full susceptibility to penicillin whereas the strains related to the C . jeikeium type strain are resistant to or only moderately susceptible to penicillin . No genomic relationship was found between C . jeikeium-related strains and other lipophilic coryneforms, identified as Corynebacterium accolens or Corynebacterium group G or F . Our study demonstrates the necessity to perform the fructose fermentation test or respiratory-type test for the correct identification of lipophilic coryneforms as C . jeikeium . Although these strains show genomic diversity at the species level, in a practical aspect, biochemical properties as well as antimicrobial susceptibility may allow the classification of such isolates in this single taxon. J Clin Pathol, 1994 Aug, 47(8), 756 - 9 Evaluation of API Coryne system for identifying coryneform bacteria; Soto A et al.; AIM--To identify rapidly and accurately coryneform bacteria, using a commercial strip system . METHODS--Ninety eight strains of Corynebacterium species and 62 additional strains belonging to genera Erysipelothrix, Oerskovia, Rhodococcus, Actinomyces, Archanobacterium, Gardnerella and Listeria were studied . Bacteria were identified using conventional biochemical tests and a commercial system (API-Coryne, BioMerieux, France) . Fresh rabbit serum was added to fermentation tubes for Gardnerella vaginalis isolates . RESULTS--One hundred and five out of the 160 (65.7%) organisms studied were correctly and completely identified by the API Coryne system . Thirty five (21.8%) more were correctly identified with additional tests . Seventeen (10.6%) organisms were not identified by the system and three (1.9%) were misidentified . CONCLUSIONS--The system was a good alternative for identification of coryneform organisms . When occasionally performed with some additional tests, this method permits reliable and rapid identification of coryneform organisms compared with conventional methods. Acta Virol, 1994 Aug, 38(4), 223 - 8 Characterization and sequence analysis of the F2 promoter from corynephage BFK20; Koptides M et al.; F2 promoter from corynephage BFK20 was isolated and characterized . It was functional in Escherichia coli and Corynebacterium glutamicum . Cloning of the F2 promoter into the pJUP05 promoter probe vector caused an increase of the neomycin phosphotransferase II specific activity . According to the Northern blot hybridization the nptII gene was expressed from the cloned F2 promoter . The apparent transcription start point in E . coli and C . glutamicum was determined . The -35 region of F2 promoter showed high similarity to that of E . coli promoter consensus sequence, but its -10 region was G+C rich and had no significant homology to that. Trop Anim Health Prod, 1994 Aug, 26(3), 168 - 74 National cross-sectional study of mastitis in dairy cattle in Jordan; Lafi S et al.; Between July 1991 and August 1992, 63 Jordanian dairy farms selected by stratified random sample were visited to identify the major causes and prevalence of intramammary infections in dairy cows . Of 773 cows examined 60% of all sampled quarters had > 283,000 cells/ml . The mean value of somatic cell count (SCC) was positively associated with age in lactations and negatively with herd size . Cows milked by bucket milking machines or in fully automatic parlours had a lower mean SCC than those milked by hand . Many management faults pertaining to milking procedure and maintenance of milking machines were noted . The most common isolate from clinical cases was Staphylococcus aureus (37.5%) . Estimates of prevalence of bacterial pathogens in intramammary infections were: coagulase negative staphylococci (16.04%), S . aureus (9.41%), Klebsiella spp . (6.17%), Corynebacterium bovis (5.35%) and Brucella melitensis (4.52%) . The results demonstrate the essential need for the development of a national mastitis control programme. Biosci Biotechnol Biochem, 1994 Aug, 58(8), 1451 - 7 Cloning of two halohydrin hydrogen-halide-lyase genes of Corynebacterium sp . strain N-1074 and structural comparison of the genes and gene products; Yu F et al.; We cloned the genes from Corynebacterium sp . strain N-1074, for two halohydrin hydrogen-halide-lyase isoenzyme (hheA and hheB), which are involved in the transformation of alpha, beta-halohydrin into the corresponding epoxide and the reverse reaction . The nucleotide sequences of 1057 base pairs including the hheA gene and of 1130 base pairs including the hheB gene were analyzed . The predicted amino acid sequence of the hheA gene product consisted of 244 residues, and the calculated molecular weight was 26,465 . The hheB gene had two sets of potential ribosome binding site and a possible initiation codon . The predicted amino acid sequences of the gene products consisted of 235 and 227 residues, and the calculated molecular weights were 26,179 and 25,236, respectively . Site-directed mutagenesis experiments strongly suggested dual translational initiation in the hheB gene . Comparison of the predicted amino acid sequences for the hheA and hheB genes found significant homology only in the carboxyl terminal region . An analysis of the upstream regions of the both hhe genes suggested the presence of putative epoxide hydrolase genes, which might be involved in further degradation of epoxide compounds. Clin Sci (Lond), 1994 Aug, 87(2), 179 - 86 Cellular localization of inducible nitric oxide synthase in experimental endotoxic shock in the rat; Cook HT et al.; 1 . Endotoxin induces a shock-like syndrome with increased nitric oxide synthesis . To clarify the cellular source of NO in endotoxic shock we used immunohistochemistry and in situ hybridization to localize inducible NO synthase in rats given lipopolysaccharide or Corynebacterium parvum and lipopolysaccharide . Immunohistochemistry was carried out with an antibody raised against a synthetic peptide of mouse macrophage NO synthase . In situ hybridization was performed with 35S-labelled oligonucleotide probes corresponding to cDNA sequences common to mouse macrophage inducible NO synthase and rat vascular smooth inducible NO synthase . Monocytes and macrophages were identified by immunohistochemistry with the mouse monoclonal antibody ED1 . 2 . After lipopolysaccharide alone, the major site of NO synthase induction was monocytes and macrophages in multiple organs, principally liver and spleen . Bronchial, bile duct, intestinal and bladder epithelium and some hepatocytes also expressed inducible NO synthase . Expression peaked at 5 h and had returned to normal by 12 h except in spleen . 3 . After priming with C . parvum, lipopolysaccharide led to a similar distribution of inducible NO synthase as lipopolysaccharide alone, but in addition there was more prominent hepatocyte staining, staining in macrophage granulomas in the liver and inducible NO synthase was present in some endothelial cells in the aorta . 4 . These findings provide a direct demonstration of the cellular localization of inducible NO synthase after lipopolysaccharide. Microbiology, 1994 Aug, 140 ( Pt 8), 1817 - 28 Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase; Eikmanns BJ et al.; Citrate synthase catalyses the initial reaction of the citric acid cycle and can therefore be considered as the rate-controlling enzyme for the entry of substrates into the cycle . In Corynebacterium glutamicum, the specific activity of citrate synthase was found to be independent of the growth substrate and of the growth phase . The enzyme was not affected by NADH or 2-oxoglutarate and was only weakly inhibited by ATP (apparent Ki = 10 mM) . These results suggest that in C . glutamicum neither the formation nor the activity of citrate synthase is subject to significant regulation . The citrate synthase gene, gltA, was isolated, subcloned on plasmid pJC1 and introduced into C . glutamicum . Relative to the wild-type the recombinant strains showed six- to eightfold higher specific citrate synthase activity . The nucleotide sequence of a 3007 bp DNA fragment containing the gltA gene and its flanking regions was determined . The predicted gltA gene product consists of 437 amino acids (M(r) 48,936) and shows up to 49.7% identity with citrate synthase polypeptides from other organisms . Inactivation of the chromosomal gltA gene by gene-directed mutagenesis led to absence of detectable citrate synthase activity and to citrate (or glutamate) auxotrophy, indicating that only one citrate synthase is present in C . glutamicum . Transcriptional analysis by Northern (RNA) hybridization and primer extension experiments revealed that the gltA gene is monocistronic (1.45 kb mRNA) and that its transcription initiates at two consecutive G residues located 121 and 120 bp upstream of the translational start. Gene, 1994 Jul 22, 145(1), 69 - 73 Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum; Schafer A et al.; Here we describe small mobilizable vectors based on the Escherichia coli plasmids pK18 and pK19 . We combined the useful properties of the pK plasmids (e.g., multiple cloning site, lacZ alpha fragment, sequencing with M13 primers) with the broad-host-range transfer machinery of plasmid RP4 and a modified sacB gene from Bacillus subtilis . The new pK derivatives can be transferred by RP4-mediated conjugation into a wide range of Gram- and Gram+ bacteria, and should facilitate gene disruption and allelic exchange by homologous recombination . As an application example, the generation of a defined deletion of the hom-thrB genes in the chromosome of the Gram+ bacterium Corynebacterium glutamicum is presented. Appl Environ Microbiol, 1994 Jul, 60(7), 2501 - 7 Structural and functional analysis of pyruvate kinase from Corynebacterium glutamicum; Jetten MS et al.; Pyruvate kinase activity is an important element in the flux control of the intermediate metabolism . The purified enzyme from Corynebacterium glutamicum demonstrated a marked sigmoidal dependence of the initial rate on the phosphoenolpyruvate concentration . In the presence of the negative allosteric effector ATP, the phosphoenolpyruvate concentration at the half-maximum rate (S0.5) increased from 1.2 to 2.8 mM, and cooperation, as expressed by the Hill coefficient, increased from 2.0 to 3.2 . AMP promoted opposite effects: the S0.5 was decreased to 0.4 mM, and the enzyme exhibited almost no cooperation . The maximum reaction rate was 702 U/mg, which corresponded to an apparent kcat of 2,540 s-1 . The enzyme was not influenced by fructose-1,6-diphosphate and used Mn2+ or Co2+ as cations . Sequence determination of the C . glutamicum pyk gene revealed an open reading frame coding for a polypeptide of 475 amino acids . From this information and the molecular mass of the native protein, it follows that the pyruvate kinase is a tetramer of 236 kDa . Comparison of the deduced polypeptide sequence with the sequences of other bacterial pyruvate kinases showed 39 to 44% homology, with some regions being very strongly conserved. Appl Environ Microbiol, 1994 Jul, 60(7), 2494 - 500 Cloning of the pyruvate kinase gene (pyk) of Corynebacterium glutamicum and site-specific inactivation of pyk in a lysine-producing Corynebacterium lactofermentum strain; Gubler M et al.; The pyruvate kinase gene pyk from Corynebacterium glutamicum was cloned by applying a combination of PCR, site-specific mutagenesis, and complementation . A 126-bp DNA fragment central to the C . glutamicum pyk gene was amplified from genomic DNA by PCR with degenerate oligonucleotides as primers . The cloned DNA fragment was used to inactivate the pyk gene in C . glutamicum by marker rescue mutagenesis via homologous recombination . The C . glutamicum pyk mutant obtained was unable to grow on minimal medium containing ribose as the sole carbon source . Complementation of this phenotype by a gene library resulted in the isolation of a 2.8-kb PstI-BamHI genomic DNA fragment harboring the C . glutamicum pyk gene . Multiple copies of plasmid-borne pyk caused a 20-fold increase of pyruvate kinase activity in C . glutamicum cell extracts . By using large internal fragments of the cloned C . glutamicum gene, pyk mutant derivatives of the lysine production strain Corynebacterium lactofermentum 21799 were generated by marker rescue mutagenesis . As determined in shake flask fermentations, lysine production in pyk mutants was 40% lower than that in the pyk+ parent strain, indicating that pyruvate kinase is essential for high-level lysine production . This finding questions an earlier hypothesis postulating that redirection of carbon flow at the phosphoenol pyruvate branch point of glycolysis through elimination of pyruvate kinase activity results in an increase of lysine production in C . glutamicum and its close relatives. Acta Obstet Gynecol Scand, 1994 Jul, 73(6), 460 - 4 Uro-genital microbial colonization and threatening preterm delivery; Svare J et al.; OBJECTIVE . To examine whether there is a relationship between the uro-genital microbial colonization and threatening preterm delivery . STUDY DESIGN . The microflora in the urine and endocervix was studied in 43 women with preterm labor, 45 women with preterm premature rupture of the membranes (PPROM) and 80 normal pregnant women at 26-34 weeks of gestation . Amniotic fluid was examined in 20 of the patients with preterm labor . Data were analyzed by Fisher's exact test (two-tailed) . RESULTS . The microflora in the urine was not significantly different in patients with preterm labor, PPROM and normal pregnant women . Compared with normal pregnant women, patients with preterm labor had significantly lower prevalences of corynebacteria (p < 0.05) and coagulase-negative staphylococci (p < 0.01) in the cervix, while patients with PPROM had significantly lower prevalences of lactobacilli (p < 0.05) and coagulase-negative staphylococci (p < 0.05) in the cervix . Positive amniotic fluid cultures were detected in three of the 20 patients with preterm labor who underwent transabdominal amniocentesis . Evidence of ascending colonization was found in two of these cases . CONCLUSIONS . The microbial colonization of the urine was not associated with threatening preterm delivery . Reduced prevalences of lactobacilli, corynebacteria and coagulase-negative staphylococci in the cervix were associated with threatening preterm delivery. Zh Mikrobiol Epidemiol Immunobiol, 1994 Jul-Aug, (4), 24 - 8 {The cloning and expression of the phospholipase D gene from Corynebacterium pseudotuberculosis}; Dmitriev AV et al.; In a number of consecutive gene engineering operations a DNA fragment having a size of about 2.8 kb was cloned in Escherichia coli by means of Blue-script II SK+ used as vector . The insert contained pld gene coding the synthesis of phospholipase D, one of the key factors of C . pseudotuberculosis virulence . The stable and active expression of this gene in E.coli was achieved . High phospholipase A activity was accumulated in the periplasmatic space . The molecular weight of the synthesized protein was 31 kD . The product obtained by gene engineering methods was found to possess the biological activity of the natural product: it induced the hemolysis of sheep red blood cells in the presence of equi factor of Rhinococcus equi and inhibited the hemolytic activity of Staphylococcus aureus beta-hemolysin (phospholipase C) . The pld gene cloned in these experiments differed from that of another C.pseudotuberculosis strain . Further research is underway with a view of searching for the limits of pls gene. Pathology, 1994 Jul, 26(3), 311 - 4 Seven patients with respiratory infections due to Corynebacterium pseudodiphtheriticum; Freeman JD et al.; Seven cases of lower respiratory tract infection due to Corynebacterium pseudodiphtheriticum are described . Lower respiratory tract infections with C . pseudodiphtheriticum in immunocompetent patients are usually associated with pre-existing chronic pulmonary disease, and are sometimes associated with endotracheal intubation . Antimicrobial susceptibility testing of these isolates showed uniform sensitivity to penicillin and variable results to erythromycin. J Clin Microbiol, 1994 Jul, 32(7), 1825 - 6 Improved enrichment broth for cultivation of fastidious organisms; Cartwright CP et al.; An enrichment broth developed in our laboratory, fastidious broth (FB), was compared with two commercially available broth media, supplemented thioglycolate broth and enriched eugonic broth . FB supported the growth of a number of organisms that were not cultivatable in either of the other two media, including Corynebacterium jeikeium, Haemophilus influenzae, Neisseria gonorrhoeae, and Streptococcus pneumoniae . In addition, for several organisms that were able to grow in all three broths, including Neisseria meningitidis, Nocardia asteroides, and Actinomyces spp., both the time of incubation and the starting inoculum necessary to enable detection of growth were decreased significantly by using FB. J Clin Microbiol, 1994 Jul, 32(7), 1705 - 9 Production of extracellular slime by coryneforms colonizing hydrocephalus shunts; Bayston R et al.; Corynebacterium spp . are responsible for an important minority of cases of colonization of cerebrospinal fluid shunts used for the treatment of hydrocephalus . In common with coagulase-negative staphylococci, they present a serious therapeutic problem because they are often resistant to multiple antibiotics . We studied the morphologies of coryneforms in colonized hydrocephalus shunts removed from patients and observed extracellular slime similar in appearance to that seen in coagulase-negative staphylococci . We also studied a series of such isolates from other cases of hydrocephalus shunt colonization using an established laboratory model and consistently observed slime production in these shunts as well . We propose that this might be a further reason for failure to eradicate these organisms without shunt removal as well as a factor in their pathogenesis in device-related infections. J Appl Bacteriol, 1994 Jul, 77(1), 49 - 53 Fungicin M4: a narrow spectrum peptide antibiotic from Bacillus licheniformis M-4; Lebbadi M et al.; The strain Bacillus licheniformis M-4 produces a 3.4 kDa hydrophilic peptide with antifungal activity, named fungicin M4 . Analysis of the purified peptide shows that it contains the amino acids Glu (8), Arg (5), Pro (4), Tyr (8), Val (3), Met (2) and Orn (4) . Its inhibitory spectrum is restricted to Microsporum canis CECT 2797, Mucor mucedo CECT 2653, Mucor plumbeus CCM 443, Sporothrix schenckii CECT 2799, Bacillus megaterium and Corynebacterium glutamicum CECT 78 . Fungicin M4 exerts biocidal activity on liquid cultures of Sporothrix schenckii CECT 2799. Gesundheitswesen, 1994 Jul, 56(7), 380 - 2 {Detection by molecular biology techniques of diphtheria toxin production in corynebacteria}; Weig M et al.; The ability to secrete a potent protein-toxin called diphtheria toxin (DT) is of outstanding importance for the outcome of diphtheria epidemics . Rapid molecular-biological screening methods are shown that can improve the differentiation between toxinogenic and non-toxinogenic Corynebacterium diphtheriae strains . The recently reported infection with an endemic, putative toxinogenic Corynebacterium diphtheriae strain in Lower Saxony is discussed. Diagn Microbiol Infect Dis, 1994 Jul, 19(3), 171 - 3 Problems in minimum inhibitory concentration determinations in coryneform organisms . Comparison of an agar dilution and the Etest; Zapardiel J et al.; The antimicrobial susceptibility of 159 coryneform organisms was determined by an agar dilution and Etest methods . Overall, the correlation between minimum inhibitory concentrations obtained by both techniques was good (> or = 0.09) for most antibiotics and organisms although the essential agreement ranged from 59% to 88.3% . Most organisms were equally categorized (sensitive, intermediate, or resistant) by both methods with only 0.2%, 0.4%, and 3.5% of very major, major or minor discrepancies, respectively . Such percentages dropped significantly when discrepant strains were retested . The correlation was specially good for Corynebacterium jeikeium and Corynebacterium urealyticum. Eur J Clin Microbiol Infect Dis, 1994 Jul, 13(7), 600 - 4 Incidence and characteristics of urinary tract infections caused by Corynebacterium urealyticum (Corynebacterium group D2); Nebreda-Mayoral T et al.; The incidence and characteristics of urinary tract infections caused by Corynebacterium urealyticum were studied prospectively in 20,766 urine samples . Corynebacterium urealyticum was isolated from 67 samples (0.32%) . Twenty-four percent of the patients from whom Corynebacterium urealyticum was isolated showed mild symptoms and had no risk factors other than prolonged hospitalization and previous antibiotic treatment . Sixty percent of the patients had urinary tract-related symptoms . The main risk factors were underlying urinary tract disease, antibiotic treatment, prolonged hospitalization and urological manipulation . Patients with antimicrobial treatment had a favourable clinical course, with the exception of two patients with encrusted cystitis. Biochem Biophys Res Commun, 1994 Jun 30, 201(3), 1255 - 62 A sequence from a tryptophan-hyperproducing strain of Corynebacterium glutamicum encoding resistance to 5-methyltryptophan; Heery DM et al.; A cloned DNA fragment containing the trp gene cluster from the tryptophan-hyperproducing strain Corynebacterium glutamicum ATCC21850 was found to increase the resistance of Escherichia coli to the tryptophan analogs 5-methyltryptophan and 6-fluorotryptophan . A sequence sufficient to mediate resistance to 5-methyltryptophan in E . coli was mapped to a 582-bp sequence located immediately upstream of the C . glutamicum trp operon . The equivalent fragment from the related wild type strain C . glutamicum AS019 was found to contain sequence differences at two positions and had no effect on the sensitivity of E . coli to 5-methyltryptophan. J Bacteriol, 1994 Jun, 176(12), 3474 - 83 Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme; Reinscheid DJ et al.; Isocitrate lyase is a key enzyme in the glyoxylate cycle and is essential as an anaplerotic enzyme for growth on acetate as a carbon source . It is assumed to be of major importance in carbon flux control in the amino acid-producing organism Corynebacterium glutamicum . In crude extracts of C . glutamicum, the specific activities of isocitrate lyase were found to be 0.01 U/mg of protein after growth on glucose and 2.8 U/mg of protein after growth on acetate, indicating tight regulation . The isocitrate lyase gene, aceA, was isolated, subcloned, and characterized . The predicted gene product of aceA consists of 432 amino acids (M(r), 47,228) and shows up to 57% identity to the respective enzymes from other organisms . Downstream of aceA, a gene essential for thiamine biosynthesis was identified . Overexpression of aceA in C . glutamicum resulted in specific activities of 0.1 and 7.4 U/mg of protein in minimal medium containing glucose and acetate, respectively . Inactivation of the chromosomal aceA gene led to an inability to grow on acetate and to the absence of any detectable isocitrate lyase activity . Isocitrate lyase was purified to apparent homogeneity and subjected to biochemical analysis . The native enzyme was shown to be a tetramer of identical subunits, to exhibit an ordered Uni-Bi mechanism of catalysis, and to be effectively inhibited by 3-phosphoglycerate, 6-phosphogluconate, phosphoenolpyruvate, fructose-1,6-bisphosphate, and succinate. J Leukoc Biol, 1994 Jun, 55(6), 711 - 8 The cooperative effects of TNF-alpha and IFN-gamma are determining factors in the ability of IL-10 to protect mice from lethal endotoxemia; Smith SR et al.; Recent studies have demonstrated that interleukin-10 (IL-10) has the capacity to protect mice from the lethal effects of endotoxin . In this investigation, we have examined the ability of IL-10 to protect both normal mice and Corynebacterium parvum-primed mice against endotoxin lethality . In the overwhelming majority of experiments, recombinant murine IL-10 (rMuIL-10) and recombinant human IL-10 (rHuIL-10) did not protect normal BALB/cJ mice from lipopolysaccharide (LPS)-induced lethality at doses up to 10 micrograms/mouse . Despite their inability to protect, both IL-10 preparations were highly effective in preventing the increase in serum tumor necrosis factor alpha (TNF-alpha) that occurred in response to the lethal dose of LPS . Moreover, a neutralizing antibody against TNF-alpha gave only partial protection when administered alone to BALB/cJ mice . Treatment with a combination of neutralizing antibodies against TNF-alpha and interferon-gamma (IFN-gamma) resulted in complete protection . In contrast to BALB/cJ mice, normal BDF1 mice were protected from lethal endotoxemia by treatment with both rMuIL-10 and rHuIL-10 . However, IL-10 did not protect C . parvum-primed BDF1 against LPS lethality even though it caused a reduction in the LPS-induced serum TNF-alpha response in C . parvum-primed mice as well as in normal BDF1 mice . Neutralizing antibodies against TNF-alpha and IFN-gamma were protective when administered alone to normal BDF1 mice, as previously demonstrated in C . parvum-primed mice . These findings suggest that lethal endotoxemia is a result of the cooperative activities of TNF-alpha and IFN-gamma in normal mice of the BALB/cJ and BDF1 strains as well as in C . parvum-primed BDF1 mice . IL-10 appears to be less effective in protecting mice from lethal endotoxemia when cooperation between IFN-gamma and TNF-alpha is facilitated by high-level production of the cytokines as in C . parvum-primed mice or when there is evidence of strong synergy between them as in normal BALB/cJ mice. J Urol, 1994 Jun, 151(6), 1636 - 7 Corynebacterium induced urethral incrustation; Park JM et al.; To our knowledge we report the first case of extensive urethral incrustation to be caused by corynebacterium group D2 infection . The condition was treated effectively by intravenous vancomycin hydrochloride, cystolitholapaxy and urinary acidification with a urease inhibitor, acetohydroxamic acid. Infect Immun, 1994 Jun, 62(6), 2562 - 7 Identification of a novel antigen from Corynebacterium pseudotuberculosis that protects sheep against caseous lymphadenitis; Walker J et al.; A 40-kDa protein antigen from Corynebacterium pseudotuberculosis has been identified by application of a strategy that employs locally derived antibody-secreting cells (ASC) . ASC probes generated by culture of ASC obtained from the lymph node draining the site of infection showed a specificity restricted to a 40-kDa antigen . Analysis of immunoblots with sequential serum samples taken from sheep during the course of experimental primary infection with C . pseudotuberculosis also revealed the 40-kDa antigen as an early immunodominant antigen . Sheep vaccinated with two 100-micrograms doses of a 40-kDa antigen preparation in aluminium hydroxide adjuvant were protected against infection with C . pseudotuberculosis, with an 82% reduction in the proportion of infected sheep and a 98% reduction in lung lesions . Sera from vaccinated sheep exhibited a strong response only to the 40-kDa antigen on immunoblots . These results strongly suggest that the 40-kDa antigen plays a major role in immunity to caseous lymphadenitis. Antimicrob Agents Chemother, 1994 Jun, 38(6), 1439 - 41 Comparative in vitro activities of new quinolones against coryneform bacteria; Martinez-Martinez L et al.; The in vitro activities of eight quinolones against 115 coryneform bacteria (20 Corynebacterium jeikeium, 15 Corynebacterium minutissimum, 15 Corynebacterium striatum, 25 Corynebacterium urealyticum, 10 Corynebacterium xerosis, 10 Corynebacterium group ANF-1, 10 Corynebacterium group 12, and 10 Listeria monocytogenes) were determined . The MICs of ciprofloxacin, ofloxacin, and sparfloxacin for 90% of C . jeikeium, C . urealyticum, and C . xerosis isolates tested were > 16 micrograms/ml . Those of BAY Y 3118 and clinafloxacin against these species were 0.5 and 1 to 2 micrograms/ml, respectively . The MICs for 90% of all 115 strains tested were 0.5 microgram/ml for BAY Y 3118, 1 microgram/ml for clinafloxacin, 2 micrograms/ml for E-5068, 4 micrograms/ml for E-5065, and > 16 micrograms/ml for ciprofloxacin, ofloxacin, sparfloxacin, and E-4868. Can J Microbiol, 1994 Jun, 40(6), 508 - 12 Comparison of thermostable macromolecular antigens from leprosy-associated bacteria; Baelden MC et al.; Three types of bacteria are associated with leprosy: Mycobacterium leprae, leprosy-derived corynebacteria (LDC), and armadillo-derived mycobacteria (ADM) . The immunological relationships between these three types of bacteria and Mycobacterium bovis BCG, used as a reference, were determined by cross-immunoelectrophoresis . When compared with the reference, cross-reactions were observed with a variable number of antigens: 2 in the case of strain LDC 15, 4 with M . leprae, and from 1 to 10 in the case of the ADM, depending on their subgroup . Next, thermostable macromolecular antigens (TMAs), the major cross-reactive antigens of leprosy-associated bacteria, were compared by anti-TMA antibody ELISA tests . The LDC TMAs displayed high cross-reactivity between the subgroups and lower cross-reactivity with the TMAs of M . bovis BCG . Evidence for the presence of a species-specific moiety in TMA of the different LDC was obtained by using depleted anti-TMA antisera . Western blot analysis revealed the presence of many proteins in the TMAs of LDC and M . bovis BCG, some of them being species-specific and other cross-reactive. FEMS Microbiol Lett, 1994 Jun 1, 119(1-2), 137 - 45 Nucleotide sequence of the gene encoding the Corynebacterium glutamicum mannose enzyme II and analyses of the deduced protein sequence; Lee JK et al.; The complete nucleotide sequence of the gene encoding the Corynebacterium glutamicum mannose enzyme II (EIIMan) was determined . The gene consisted of 2052 base pairs encoding a protein of 683 amino acid residues; the molecular mass of the protein subunit was calculated to be 72570 Da . The N-terminal hydrophilic domain of EIIMan showed 39.7% homology with a C-terminal hydrophilic domain of Escherichia coli glucose-specific enzyme II (EIIGlc) . Similar homology was shown between the C-terminal sequence of EIIMan and the E . coli glucose-specific enzyme III (EIIIGlc), or the EIII-like domain of Streptococcus mutans sucrose-specific enzyme II . Sequence comparison with other EIIs showed that EIIMan contained residues His-602 and Cys-28 which were homologous to the potential phosphorylation sites of EIIIGlc, or EIII-like domains, and hydrophilic domains (IIB) of several EIIs, respectively. Am J Physiol, 1994 Jun, 266(6 Pt 1), G1004 - 10 Nitric oxide and prostaglandins interact to prevent hepatic damage during murine endotoxemia; Harbrecht BG et al.; Nitric oxide (NO) and prostaglandins (PG) both possess the ability to induce vasodilatation and prevent the aggregation of platelets . The synthesis of these substances is increased following in vivo lipopolysaccharide (LPS) infusion, but their function during sepsis is incompletely understood . We studied the role of NO and PG in a murine model of chronic hepatic inflammation (Corynebacterium parvum injection), which is known to progress to sudden hepatic necrosis after LPS injection . NO synthesis, which is induced in hepatocytes by C . parvum treatment and in nonparenchymal cells by LPS treatment, was inhibited using NG-monomethyl-L-arginine (L-NMMA) . High-dose aspirin (ASA) was used to block PG synthesis . Treatment with L-NMMA or ASA alone, in the absence of LPS, resulted in no increase in hepatic injury . C . parvum-treated mice that received both L-NMMA and ASA without LPS developed marked hepatic damage as reflected by increased hepatocellular enzyme release (aspartate aminotransferase and L-ornithine carbamoyl-transferase) . Marked hepatic damage was seen after LPS administration, and ASA pretreatment alone had no effect on the LPS-induced hepatic injury, whereas L-NMMA markedly increased the hepatic damage . The combination of L-NMMA and ASA after LPS resulted in the greatest hepatocellular enzyme release, characterized histologically by intravascular thrombosis with diffuse infarction and necrosis . Simultaneous treatment with either PGI2 or L-arginine partially prevented this injury . These data demonstrate that NO and PG function synergistically to maintain hepatocellular integrity; thus increased synthesis of these mediators protects the liver from the pathophysiological effects of LPS in this model. J Trop Med Hyg, 1994 Jun, 97(3), 189 - 91 Corynebacterium diphtheriae endocarditis in an adult with congenital heart disease: a case report; Lin RV et al.; A rare case of septicaemia and endocarditis due to Corynebacterium diphtheriae is presented . The patient had underlying congenital heart disease and died despite high-dose penicillin therapy . The implications for diagnosis and identification and some public health issues brought to the fore by the case are discussed. Bioorg Med Chem, 1994 Jun, 2(6), 535 - 42 Stereoselective epoxidation of 2,2-dimethyl-2H-1-benzopyran-6-carbonitrile; Patel RN et al.; The chiral intermediate (3S,4R)-trans-3,4-dihydro-3,4-dihydroxy-2,2-dimethyl- 2H-1-benzopyran-6-carbonitrile {(+)-trans diol 3} was made by the stereoselective microbial epoxidation of 2,2-dimethyl-2H-1-benzopyran-6-carbonitrile 1 . This compound is a potential intermediate for the total synthesis of potassium-channel openers . Several microbial cultures were found which catalyzed the transformation of 1 to the corresponding (3S,4S)-epoxide 2 and (+)-trans diol 3 . The two best cultures, Corynebacterium sp . SC 13876 and Mortierella ramanniana SC 13840 gave reaction yields of 32 M% and 67.5 M% and optical purities of 88 and 96%, respectively, for (+)-trans diol 3 . A single-stage process (fermentation-epoxidation) for the biotransformation of 1 was developed using Corynebacterium sp . SC 13876 and M . ramanniana SC 13840 . In a 25-L fermentor, the (+)-trans diol 3 was obtained in 38.6 M% yield with an optical purity of 90% using Corynebacterium SC 13876 . The reaction yield of 60.7 M% and optical purity of 92.5% were obtained for (+)-trans diol 3 using M . ramanniana SC 13840 . A two-stage process for the preparation of (+)-trans diol 3 was also developed using a 3 L cell-suspension (10% w/v, wet cells) of M . ramanniana SC 13840 . The reaction was carried out in a 5-L Bioflo fermentor . The concentration of substrate 1 was 2 g L-1 with glucose present at 10 g L-1 . After 48 h, (+)-trans diol 3 was obtained in 76 M% yield with an optical purity of 96%.(ABSTRACT TRUNCATED AT 250 WORDS) Mol Microbiol, 1994 Jun, 12(6), 921 - 30 Targeted mutagenesis of the phospholipase D gene results in decreased virulence of Corynebacterium pseudotuberculosis; McNamara PJ et al.; The chromosomal gene encoding the phospholipase D from Corynebacterium pseudotuberculosis (biovar ovis) isolate Whetten 1 was replaced with an allele containing a nonsense mutation . The virulence of the mutant strain (W1.31r1) and the isogenic parental strain were then compared by inoculation of goats . The wild-type strain caused abscessation at the site of infection, which then spread to the regional lymph node, while W1.31r1 had a reduced ability to establish a primary infection and was incapable of dissemination . Our results confirm that phospholipase D is a virulence determinant of C . pseudotuberculosis that increases the persistence and spread of the bacteria within the host. Microb Pathog, 1994 Jun, 16(6), 387 - 99 Herpes simplex virus type 1 replication and IL-1 beta gene expression in mouse peritoneal macrophages activated in vivo by an attenuated Salmonella typhimurium vaccine or Corynebacterium parvum; Wu L et al.; Activated macrophages (M phi) from mice given Salmonella typhimurium or Corynebacterium parvum were compared with resident peritoneal macrophages at the molecular level for permissiveness for herpes simplex virus type 1 (HSV-1) replication and for expression of interleukin-1 beta (IL-1 beta) . Peritoneal macrophages were harvested from mice injected 7 days previously with live, avirulent S . typhimurium (Sal-PM phi) or heat-killed C . parvum (CP-PM phi) and infected with HSV-1 in vitro . Both Sal-PM phi and CP-PM phi were activated as evidenced by characteristic changes in an ectoenzyme, by increased permissiveness for infectious virus production and viral cytopathic effect, and by induction of IL-1 beta mRNA . Analysis at the molecular level revealed that both types of activated M phi demonstrated increased patterns of HSV-1 immediate-early gene expression and viral DNA replication as compared with resident cells . A novel finding was that viral infection reduced IL-1 beta mRNA in both types of activated M beta . This observation has implications for the efficacy of Salmonella vaccines given in proximity to HSV-1 infection and for potential deleterious effects of HSV-1 infection in immunosuppressed patients receiving immunotherapy. Infect Immun, 1994 May, 62(5), 1600 - 8 Characterization of mutations that inactivate the diphtheria toxin repressor gene (dtxR); Wang Z et al.; The diphtheria toxin repressor (DtxR) is an iron-dependent regulator of diphtheria toxin production and iron uptake in Corynebacterium diphtheriae . It is activated in vitro by divalent metal ions including Fe2+, Cd2+, Co2+, Mn2+, Ni2+, and Zn2+ . We characterized 20 different mutations in dtxR induced by bisulfite mutagenesis, 18 of which caused single-amino-acid substitutions in DtxR and two of which were chain-terminating mutations . Six of the amino acid replacements were clustered between residues 39 and 52 in a predicted helix-turn-helix motif that exhibits homology with several other repressors and is identified as the putative DNA-binding domain of DtxR . Three substitutions occurred within a predicted alpha-helical region with the sequence His-98-X3-Cys-102-X3-His-106 that resembles metal-binding motifs in several other proteins and is identified as the putative metal-binding site of DtxR . Several purified variants of DtxR with decreased repressor activity failed to bind in gel retardation assays to DNA fragments that contained the tox operator . A quantitative assay for binding of DtxR to 63Ni2+ was also developed . Scatchard analysis revealed that DtxR has a single class of high-affinity 63Ni(2+)-binding sites with a Kd of 2.11 x 10(-6) M and a maximum binding capacity of approximately 1.2 atoms of Ni2+ per DtxR monomer . The P39L, T40I, T44I, and R47H variants of DtxR exhibited normal to slightly decreased 63Ni(2+)-binding activity, but H106Y, which has an amino acid substitution in the presumed metal-binding domain, exhibited markedly decreased 63Ni(2+)-binding activity. Blood, 1994 May 1, 83(9), 2698 - 706 Photochemical inactivation of pathogenic bacteria in human platelet concentrates; Lin L et al.; Platelet concentrates (PC) may be infrequently contaminated with low levels of bacteria that can cause septicemia and death in patients receiving transfusion therapy . We evaluated the efficacy of a photochemical decontamination (PCD) technique using 8-methoxypsoralen (8-MOP) and long wavelength UV light (UVA) to inactivate bacteria in standard therapeutic PC . Twelve phylogenetically distinct pathogenic bacteria, 5 gram-positive and 7 gram-negative organisms, were seeded into PC to a final challenge dose ranging from 10(5) to 10(7) colony-forming units (CFU)/mL . Contaminated PC were treated with 8-MOP (5 micrograms/mL) and 5 J/cm2 of UVA, a PCD treatment regimen found to adequately preserve in vitro platelet function . Greater than 10(5) CFU/mL of all 5 gram-positive (Staphylococcus aureus, Streptococcus epidermidis, Streptococcus pyogenes, Listeria monocytogenes, and Corynebacterium minutissimum) and 2 of the gram-negative (Escherichia coli and Yersinia enterocolitica) organisms were inactivated . The remaining 5 gram-negative organisms were more resistant, with less than 10(1) to 10(3.7) CFU/mL inactivated under these conditions . The inactivation efficiency for this resistant group of gram-negative organisms was improved when PC were resuspended in a synthetic storage medium with reduced plasma protein concentration (15%) and an increased 8-MOP concentration (23.4 micrograms/mL) . Illumination with 3 J/cm2 of UVA in this system inactivated greater than 10(5) CFU/mL of 4 resistant gram-negative organisms (Salmonella choleraesuis, Enterobacter cloacae, Serratia marcescens, and Klebsiella pneumoniae) and 10(4.1) CFU/mL of the most resistant gram-negative organism (Pseudomonas aeruginosa) . This level of PCD treatment did not adversely affect in vitro platelet function . These results demonstrate that PCD using 8-MOP (5 to 23.4 micrograms/mL) effectively inactivated high levels of pathogenic bacteria in PC with adequate preservation of in vitro platelet properties. J Infect, 1994 May, 28(3), 323 - 5 Cerebrospinal fluid shunt infection due to Corynebacterium xerosis; Gaskin PR et al.; We report the case of a neonate who developed ventriculitis following insertion of a ventriculoperitoneal shunt . Corynebacterium xerosis was isolated from CSF and from the tip of the catheter after it was removed . The isolate was resistant to multiple antibiotics, but the infant responded to treatment with vancomycin. Infect Control Hosp Epidemiol, 1994 May, 15(5), 315 - 8 Quantitative skin cultures at potential catheter sites in neonates; Bertone SA et al.; OBJECTIVE: To identify and quantify the bacterial and fungal flora present at body sites used for vascular catheterization of infants in a neonatal intensive care unit . DESIGN: Quantitative skin cultures were obtained from a group of neonatal patients to determine the bacterial flora found on the skin at four sites . Quantitative cultures of the jugular, subclavian, umbilical, and femoral sites were obtained on 50 infants, ranging in age from 2 days to 3 months old . SETTING: The neonatal intensive care unit of St . Christopher's Hospital for Children, a university-affiliated tertiary care children's hospital . RESULTS: Colony counts ranged from 0 to 10(6) colony-forming units/10 cm2 . Types of organisms found were consistent with other published studies and included coagulase-negative staphylococci, Staphylococcus aureus, yeast, aerobic gram-negative rods, Enterococcus species, Corynebacterium species, and alpha-hemolytic streptococci . There was a significantly higher mean colony count at the combined jugular/femoral sites versus the subclavian site (P < 0.01) and umbilical site (P < 0.05) . Mean colony counts did not differ significantly between the jugular and femoral site, or between the subclavian and umbilical site . The umbilical site was more likely to be colonized with aerobic gram-negative rods, Enterococcus species, and yeast, while the subclavian had coagulase-negative staphylococci as the predominant organism . The jugular and femoral sites demonstrated a higher colony count of aerobic gram-negative rods, Enterococcus species and yeast than the other sites . If central venous catheters need to be in place for extended periods of time, placement at a site with lower bacterial densities on the skin may help minimize catheter-associated infections . This study supports the subclavian as the preferred site. J Clin Microbiol, 1994 May, 32(5), 1395 - 6 Prevalence of Corynebacterium urealyticum in urine specimens collected at a university-affiliated medical center; Ryan M et al.; Corynebacterium urealyticum (formerly Corynebacterium group D2) has been implicated as a cause of alkaline-encrusted cystitis and urinary tract struvite calculi . Despite preselecting urine specimens with neutral and alkaline pHs and using prolonged incubation on a selective medium, isolation of this organism was rarely observed in a population of hospitalized patients . We do not recommend routine cultures for this organism unless the urine is alkaline and struvite crystals, leukocytes, and erythrocytes are present. Appl Environ Microbiol, 1994 May, 60(5), 1641 - 5 Expression of ovine gamma interferon in Escherichia coli and Corynebacterium glutamicum; Billman-Jacobe H et al.; Bacteria of two species, Escherichia coli and Corynebacterium glutamicum, were used as hosts to express recombinant ovine gamma interferon as a fusion protein with glutathione S-transferase . The recombinant gamma interferon produced by both bacteria was biologically active in vitro and was recognized by anti-gamma interferon monoclonal antibodies . E . coli produced large amounts of soluble recombinant protein which could be purified by a simple affinity chromatography method . Only a small fraction of the recombinant protein made by C . glutamicum was recovered by this method . Expression of recombinant protein in C . glutamicum was unstable but could be controlled by increased regulation of the tac promoter . Both hosts expressed ovine gamma interferon at high levels, with the recombinant protein making up a significant proportion of the cellular protein content. Mikrobiologiia, 1994 May-Jun, 63(3), 431 - 8 {Effect of antiseptics and redox-cycling agents on Corynebacterium ammoniagenes and related microorganisms in relation to synthesis of a new macroergic compound}; Ostrovskii DN et al.; Sublethal concentration of the antiseptic composition Desoxon-1 was shown to provoke in cells of Corinebacterium ammoniagenes in a liquid medium the biosynthesis and accumulation of a novel macroergic 2-methylbutane-1,2,3,4-tetraol-2,4-cyclopyrophosphate . This substance is also synthesized when C . ammoniagenes is cultivated in a solid agar medium supplemented with benzylviologen . Cells preloaded with the new cyclopyrophosphate maintain its content when treated with 4% phenol, DP-2, Desoxon-1 or boiled and heated in an autoclave . Experiments with Mycobacterium tuberculosis and BCG revealed the ability of these bacteria to grow in a medium supplemented with BV++ possibly due to ability of synthesis of a new cyclopyrophosphate which was shown to correlate with resistance toward redox-cycling drugs . Accumulation of polyphosphates in the control cells of M . tuberculosis was illustrated by 31P-NMR spectroscopy and disappearance of the polyphosphates during cultivation in a BV(++)-supplemented medium . No signal of the new cyclopyrophosphate was yet registered in cells of M . tuberculosis by 31P-NMR. Trop Anim Health Prod, 1994 May, 26(2), 74 - 8 An indirect fluorescent antibody technique for detection of anti-Dermatophilus congolensis antibodies in sheep; Hermoso De Mendoza J et al.; An indirect fluorescent antibody (IFA) technique has been developed for detection of anti-Dermatophilus antibodies in sheep . Sera from 25 bacteriologically confirmed clinically affected sheep and from 10 negative non affected lambs were used . Whole cell antigen from brain heart infusion cultures of D . congolensis was used and all sera were tested in the same way for cross-reactivity against antigens obtained from cultures of Actinomyces viscosus, Micrococcus luteus, Nocardia asteroides, and Corynebacterium pseudotuberculosis . Sera from Dermatophilus-infected sheep gave positive results with D . congolensis antigen and negative results with the antigens from other bacteria . The whole cell antigens employed were simple to prepare and easy to recognise by microscopy . Cross-reactivity was further tested using the D . congolensis culture whole cell antigen and 3 sera from sheep with bacteriologically confirmed natural infections due to Corynebacterium pseudotuberculosis, Actinomyces pyogenes and Nocardia asteroides . None of these sera showed positive reactions . The authors recommend this technique for serological surveys and research on dermatophilosis. J Neuroimmunol, 1994 May, 51(2), 199 - 208 Cell adhesion molecules on vessels during inflammation in the mouse central nervous system; Engelhardt B et al.; The expression of endothelial cell adhesion molecules (CAMs) in the central nervous system (CNS) of the mouse was examined during an inflammation induced by intracerebral injection of killed Corynebacterium parvum (C . parvum) . We showed that injection of killed C . parvum produced an inflammatory cellular infiltrate limited to the injected brain hemisphere . However, the upregulation of ICAM-1 and VCAM-1 on brain endothelium occurred starting 2 days after C . parvum injection throughout the entire CNS and was not restricted to vessels surrounded by a cellular infiltrate . In contrast to the systemic upregulation of ICAM-1 and VCAM-1, cerebral vessels located in the center of the cellular infiltrate started to express the MECA-32 antigen, suggesting an altered functional status of the endothelial cells, as this antigen is suppressed during development of the blood-brain barrier (BBB) . Binding assays performed on frozen sections of inflamed brains are consistent with an important role for endothelial VCAM-1 in the recruitment of lymphocytes during inflammation in the CNS of the mouse. Dtsch Med Wochenschr, 1994 Apr 15, 119(15), 548 - 52 {Toxic diphtheria caused by Corynebacterium ulcerans}; Hust MH et al.; A 42-year-old woman with bronchial asthma since childhood was admitted to hospital because of severe dyspnoea . Emergency bronchoscopic intubation had to be performed for life-threatening inspiratory and expiratory stridor . This demonstrated that the larynx was covered by a dirty-grey membrane and the vocal-cord gap was narrowed to a mere slit . As laryngeal diphtheria was suspected 2000 IU/kg diphtheria antitoxin was administered together with 1 mega U penicillin G four times daily intravenously . On the same day, Corynebacterium ulcerans, a very rare cause of diphtheria, was isolated from a coughed-up piece of the membrane . Toxic, massive swelling of the lymph nodes and soft tissues of the neck necessitated maintenance of an open upper airway by intubation and (later) tracheostomy for 41 days . A week later the patient was discharged without any permanent defect. J Appl Bacteriol, 1994 Apr, 76(4), 314 - 9 A variable response of degrading bacteria to phosphorus added to natural water; Ramadan MA; The effect of inorganic phosphorus (P) on the degradation of 10 mg l-1 of para-nitrophenol (PNP) or 2,4-dichlorophenoxyacetic acid (2,4-D) by three test bacteria inoculated into Nile water samples was investigated . The response of the organisms to P depended mainly on their affinity for the available P . Thus, Corynebacterium sp . at an initial density of 3.3 x 10(4) cells ml-1 readily degraded 10 mg l-1 of PNP in filter-sterilized Nile water supplemented with 22.8 mg l-1 of P . The same effect was observed when Pseudomonas cepacia was inoculated into Nile water amended with PNP and supplemented with 2.28-22.8 mg l-1 of P . The bacteria grew in Nile water and the final densities were related to the level of the added P . On the other hand, the addition of P, at concentrations ranging from 2.28 to 22.8 mg l-1, to sterile Nile water inoculated with Pseudomonas sp . and amended with 10 mg l-1 of 2,4-D did not stimulate the degradation compared with that obtained with the unsupplemented samples . The affinity of the three strains to P was demonstrated in P-deficient medium amended with PNP or 2,4-D as a sole carbon source . The pH of the medium was adjusted with 0.1 mol l-1 Tris buffer . Pseudomonas sp . at an initial density of 3.3 x 10(4) cells ml-1 degraded 10 mg l-1 of 2,4-D in non-sterile Nile water without added P . A slight enhancement of degradation was observed in water samples amended with a high concentration of P.(ABSTRACT TRUNCATED AT 250 WORDS) Hepatology, 1994 Apr, 19(4), 973 - 9 Contribution of complement-stimulated hepatic macrophages and neutrophils to endotoxin-induced liver injury in rats; Jaeschke H et al.; The role of complement as potential activator for tissue macrophages and neutrophils was investigated in an experimental model of endotoxin-induced liver injury in male Fischer rats . Injection of Salmonella enteritidis endotoxin (1 mg/kg) into Corynebacterium parvum-pretreated animals (7 mg/kg; single dose 6 days before endotoxin) resulted in severe oxidant stress, as indicated by a 37-fold increase of plasma levels of glutathione disulfide (basal concentration, 0.36 +/- 14 mumol/L), accumulation of neutrophils in the liver (600 +/- 31 neutrophils/50 high-power fields) and liver injury (plasma ALT, 1184 +/- 185 U/l; necrosis; 19% +/- 3%) 10 hr after endotoxin . The oxidant stress induced by 1 mg/kg endotoxin in the C . parvum-treated animals was always significantly higher than that in control animals receiving the same dose of endotoxin . Inhibition of complement activation with the soluble complement receptor type 1 attenuated the oxidant stress and liver injury by 50% to 65% but had no effect on hepatic neutrophil accumulation or plasma tumor necrosis factor-alpha levels . Treatment with a monoclonal antibody directed against the alpha-chain of CD11b/CD18 adhesion proteins (clone 17), which was highly effective in attenuating ischemia-reperfusion injury in the liver by reducing the number of neutrophils and functionally inactivating these cells, neither protected against parenchymal cell injury nor affected hepatic neutrophil infiltration in the C . parvum model . We conclude that reactive oxygen derived from complement-stimulated macrophages is critical for the development of liver injury in the C . parvum/endotoxin model. J Biochem (Tokyo), 1994 Apr, 115(4), 664 - 9 Cloning and sequence determination of the gene coding for the elongation factor Tu of Mycobacterium leprae; Dhandayuthapani S et al.; Elongation factor Tu (EF-Tu) plays an important role in protein biosynthesis and is susceptible to antibiotics in prokaryotes like Escherichia coli . In order to understand the primary structure of EF-Tu in the intracellular pathogenic bacterium Mycobacterium leprae, the gene (tuf gene) coding for this protein was cloned and sequenced . The gene contains a coding region of 1,188 bp with GUG as start codon . The deduced amino acid sequence has 396 amino acids with a molecular weight of 43.6 kDa . Putative GTP-binding sites are located at amino acid positions 19-24, 83-87, and 138-141 . Comparison of M . leprae EF-Tu amino acid sequence with those of M . tuberculosis, Micrococcus luteus, E . coli, and Salmonella typhimurium reveals 74-95% homology . Mitochondrial EF-Tu of Saccharomyces cerevisiae (62%) and chloroplast EF-Tu of Arabidopsis thalina (65.6%) also show strong homology with that of M . leprae . In contrast, the EF-Tu of the archaebacterium Halobacterium marismoruti exhibits relatively less homology (36.7%) . Southern hybridization of M . leprae tuf gene with genomic DNA of slow growing and fast growing mycobacteria and related species like Corynebacterium fascians and Nocardia asteroides suggests that the gene is highly conserved in these organisms. J Vet Med Sci, 1994 Apr, 56(2), 411 - 2 Protection from caseous lymphadenitis in sheep by spraying iodine tincture on shearing wounds; Serikawa S et al.; The effect of spraying shearing wounds with iodine tincture on Corynebacterium pseudotuberculosis infection in lambs was examined . The ELISA-negative lambs which had received some visible wounds during their first shearing were randomly divided into two groups: one was sprayed with iodine tincture on wounds after shearing, and the other was not . Anti-C . pseudotuberculosis toxin titers were measured by ELISA . The seroconversion ratio of the group with iodine tincture treatment 3 months after shearing was smaller than that of the untreated group (P < 0.05) . These results suggest that treatment of shearing wounds with iodine tincture is effective in the protection of C . pseudotuberculosis infection in lambs. J Clin Pathol, 1994 Apr, 47(4), 353 - 6 Polymerase chain reaction for screening clinical isolates of corynebacteria for the production of diphtheria toxin; Pallen MJ et al.; AIMS--To assess the performance of the polymerase chain reaction (PCR) when used to screen rapidly large numbers of corynebacteria for toxin production; and to determine the incidence of false positive PCR results with non-toxigenic Corynebacterium diphtheriae isolates . METHODS--Eighty seven recent British isolates of corynebacteria were assayed by PCR . All isolates were assayed from both blood and tellurite agar within a five day period . Thirty three non-toxigenic isolates of C diphtheriae from six countries were also tested by PCR and by the Elek immunodiffusion assay . RESULTS--There was complete concordance between the results of PCR and traditional methods on the recent British isolates, with one exception: an Elek positive "C ulcerans" isolate, which was PCR positive from tellurite but not from blood agar . One of the thirty three (3%) non-toxigenic isolates of C diphtheriae was PCR positive . CONCLUSIONS--These results suggest that PCR compares favourably with traditional methods for the detection of toxigenic corynebacteria and that it represents a powerful new tool in the diagnosis of an old disease. J Clin Microbiol, 1994 Apr, 32(4), 871 - 5 Mixed-effect models for predicting microbial interactions in the vaginal ecosystem; Ross RA et al.; Three statistical models that predict microbial interactions within the vaginal environment are presented . A large data set was assembled from in vivo studies describing the healthy vaginal environment, and the data set was analyzed to determine whether statistical models which would accurately predict the interactions of the microflora in this environment could be formulated . During assembly of the data set, two new variables were defined and were added to the data set, that is, cycle (sequence of menstrual cycle) and flow stage (subdivision of cycle determined by day of menstrual cycle) . Concentrations of total aerobic (includes facultative) bacteria, total anaerobic bacteria, and a Corynebacterium sp . were identified by correlation analysis as variables with significant predictors . By using a regression method with a backward elimination procedure, significant predictors of these outcome variables were identified as the concentrations of Lactobacillus spp., anaerobic Streptococcus spp., and Staphylococcus spp., respectively . For all three outcome variables, pH and flow stage were also identified as significant independent variables . Because some of the data in the data set are repeated measurements for a subject, a mixed-effect model that accounts for the random effects of repeated-measurement data fit best the data set for predicting interactions between various members of the vaginal microflora . The predictive accuracies of the three models were tested by a comparison of model-predicted outcome-variable values with actual mean in vivo outcome-variable values . From these results, we concluded that it is possible to accurately predict vaginal microflora interactions by using a mixed-effect modeling system . The application of this type of modeling strategy and its future use are discussed. Appl Environ Microbiol, 1994 Apr, 60(4), 1297 - 301 Characterization of a novel enantioselective halohydrin hydrogen-halide-lyase; Nakamura T et al.; Enzymes Ia and Ib of Corynebacterium sp . strain N-1074 exhibit halohydrin hydrogen-halide-lyase (H-lyase) activity, catalyzing the interconversion of halohydrins to epoxides and hydrogen halide . H-lyase B produced in a recombinant Escherichia coli strain carrying one of the enzyme genes of Corynebacterium sp . strain N-1074 was purified and characterized . The purified enzyme catalyzed the transformation of prochiral 1,3-dichloro-2-propanol (DCP) into R-rich epichlorohydrin (ECH) . The apparent Km values for DCP, ECH, and chloride were calculated to be 1.03, 5.00, and 4.00 mM, respectively . Maximum activity for the conversion of DCP to ECH was found at pH 8.0 to 9.0, and that for the reverse reaction was found at about pH 5.0 . H-lyase B seemed to be identical to enzyme Ib of Corynebacterium sp . strain N-1074 from the comparison of the properties of each . The properties of H-lyase B and H-lyase A, which had been previously purified from another recombinant carrying its gene from Corynebacterium sp . strain N-1074, were also compared. APMIS, 1994 Apr, 102(4), 317 - 20 Treatment of shunt-related cerebral ventriculitis due to Corynebacterium jeikeium with vancomycin administered intraventricularly . Case report; Knudsen JD et al.; A 52-year-old female with subarachnoid haemorrhage and hydrocephalus was treated with external and later internal drainage . She developed ventriculoperitoneal shunt-related ventriculitis caused by Corynebacterium jeikeium . The infection was unsuccessfully treated with intravenous vancomycin . It was controlled only after shunt removal and administration of intraventricular vancomycin as well as systemic vancomycin, rifampicin and fusidic acid . A review of the literature confirmed our experience that vancomycin given intraventricularly is well tolerated and doses can be individualized by measuring vancomycin levels in cerebrospinal fluid. Rev Argent Microbiol, 1994 Apr-Jun, 26(2), 65 - 71 {Causal agents of onychomycosis}; Canteros GE et al.; The results obtained with 307 specimens from putatively immunocompetent patients between May 1991 and May 1992 were reviewed, to determine the frequency of isolation of fungal species causing onychomycoses . Sixty eight percent of the specimen were positive for microscopic examination and/or cultures . Onychomycoses occurred with double frequency in women than in men (Table 1), and 77% of cases were diagnosed in patients aged between 30 and 70 years (Figure 1) . Out of 182 patients with positive cultures, 60% were affected by dermatophytes and 39% by yeasts; molds (Aspergillus spp.) were isolated in only two cases (Table 3) . Neither Corynebacterium spp., nor Malasezzia furfur were detected . In toe nails Trichophyton rubrum predominated over yeasts being isolated in 72.9% of the cases; yeasts other than Candida albicans were isolated in 12.3%, Trichophyton mentagrophytes in 10%, while Aspergillus spp., C . albicans and Epidermophyton floccosum in only 1.6% . On the other hand, in finger nails yeasts predominated: C . albicans was isolated in 46.7% of cases, other yeasts in 43.3%; and T . rubrum in the remaining 10% . Out of 41 isolations of yeasts other than C . albicans, 42% were C . parapsilosis, 16% Debaryomyces hansenii, 6% C . pulcherrima, 6% Trichosporum cutaneum and 6% C . famata (Figure 2). Diagn Microbiol Infect Dis, 1994 Apr, 18(4), 219 - 27 Detection of a genetic locus encoding resistance to rifampin in mycobacterial cultures and in clinical specimens; Hunt JM et al.; The polymerase chain reaction (PCR) and automated DNA sequencing were used to detect a genetic locus, rpoB, associated with rifampin resistance in Mycobacterium tuberculosis (TB) in clinical isolates and directly in clinical specimens . Primers derived from the sequence of a TB rpoB gene fragment were used to amplify DNA from bacterial and mycobacterial isolates . An rpoB-specific PCR product was obtained for five of five TB, seven of eight other mycobacterial species, Nocardia sp., Corynebacterium sp., Streptomyces sp., Actinomyces sp., and Rhodococcus sp., but not for 15 isolates (eight genera) representing usual bacterial flora . Sequence comparison of the amplified rpoB region revealed the occurrence of TB-specific "signature nucleotides" at three positions . PCR yielded amplification products for seven of 16 clinical specimens . Five of the seven contained TB-specific DNA, as well as sequences that predicted rifampin susceptibility in accord with agar dilution results . None of ten specimens that were culture negative for TB yielded TB-specific PCR products . These results with a limited number of clinical specimens demonstrate the feasibility of direct detection by PCR of rifampin-resistant TB in clinical specimens . Such testing may serve as a rapid surrogate test for multidrug-resistant TB in laboratories with PCR and automated sequencing capability. Biosci Biotechnol Biochem, 1994 Apr, 58(4), 674 - 8 Fermentative production of tryptophan by a stable recombinant strain of Corynebacterium glutamicum with a modified serine-biosynthetic pathway; Ikeda M et al.; Introduction of plasmid pKW99, which coexpresses the deregulated 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and tryptophan-biosynthetic enzymes, into tryptophan-producing Corynebacterium glutamicum KY10894 resulted in a marked increase (54%) in yield of tryptophan production (43 g/liter), but incurred two problems . One was a decline in sugar consumption at the late stage of fermentation, and the other the loss of the plasmid in the absence of selective pressure . The retarded sugar assimilation was found to be attributed to the death of cells that arose from the detrimental action of indole, the last intermediate in the tryptophan pathway, accumulated as a by-product . A chain of these events simultaneously disappeared when serine, the other substrate of the final reaction by tryptophan synthase, was added . These results indicated that a limiting supply of serine was the cause of the decline in the sugar consumption . Thus, to increase carbon flux into serine, the gene for 3-phosphoglycerate dehydrogenase (PGD), the first enzyme in the serine pathway, was cloned from wild-type C . glutamicum ATCC 31833 and joined onto pKW99 to generate pKW9901 . Strain KY10894 transformed with pKW9901 favorably consumed sugar through fermentation with accumulating little indole . Furthermore, on the basis of the observation that serine in the medium was consumed rapidly by the recombinant cells, we developed a unique plasmid stabilization system composed of KY9218 (a PGD-deficient serine-requiring strain of KY10894) and pKW9901: In its combination, cells lacking the plasmid should not proliferate in the fermentation medium which does not contain serine . Even if selective pressure was not applied, the modified strain KY9218 with pKW9901 stably maintained the plasmid during fermentation and produced 50 g/liter of tryptophan in a 61% increased yield relative to strain KY10894. Biosci Biotechnol Biochem, 1994 Apr, 58(4), 635 - 8 Cloning of the ATP phosphoribosyl transferase gene of Corynebacterium glutamicum and application of the gene to L-histidine production; Mizukami T et al.; Corynebacterium glutamicum mutants lacking ATP phosphoribosyl transferase (PRT) were selected by complementation with the Escherichia coli PRT gene . The recombinant plasmid pCH13 carrying a wild type PRT gene from C . glutamicum T106 was obtained in one of the mutants, LH13 . Transformants, LH13/pCH13 and T106/pCH13, had three times higher PRT specific activity than T106 . The plasmid pCH99 specifying the PRT, which was desensitized to feedback inhibition by L-histidine fifty-fold higher than the wild type PRT, was derived from pCH13 . L-Histidine productivity of C . glutamicum F81, was markedly decreased by pCH13, but increased twice by pCH99 . In cultivation in jar fermentors, F81/pCH99 continued to accumulate L-histidine through fermentation and yielded to the titer of 22/5 g/liter, while F81 accumulated only 11.5 g/liter due to production retardation halfway through fermentation . Moreover, F81/pCH99 had a larger production rate than F81 even in its production phase . These results indicate that the yield improvement results from amplification of the highly desensitized PRT provided by pCH99. J Am Vet Med Assoc, 1994 Mar 15, 204(6), 934 - 7 Osteomyelitis and disseminated infection caused by Corynebacterium renale in a goat; Altmaier KR et al.; A 1.5-year-old female goat was examined for recurrence of lameness involving the right forelimb . Radiography of the thorax and right scapulohumeral joint revealed a pathologic fracture of the supraglenoid tubercle, and circumscribed radiolucent lesions in the right third and fourth ribs, and the base of the spinous process of T3 . Bone scintigraphy demonstrated additional lesions in the lumbar spine and the wings of the ilium . At necropsy, disseminated infection and hematogenous osteomyelitis were diagnosed . Corynebacterium renale was cultured from the rib lesions . In food animals, osteomyelitis usually develops secondary to traumatic wounds, and members of the genus Actinomyces are frequently incriminated as the causative organism . This case is unusual because the osteomyelitis was unrelated to a traumatic wound, and the gross and microscopic lesions were reminiscent of caseous lymphadenitis, an infection caused by C pseudotuberculosis. Immunology, 1994 Mar, 81(3), 402 - 6 Induced hyporesponsiveness in rat Kupffer cells is not specific for lipopolysaccharide; Gonnella PA et al.; The phenomenon of lipopolysaccharide (LPS)-induced hyporesponsiveness has been reported to occur in macrophage cell lines and primary cells . Hyporesponsiveness was evidenced by a diminution or lack of production of tumour necrosis factor-alpha (TNF-alpha) after sequential doses of LPS . In order to characterize the hyporesponsive state in Kupffer cells, the production of TNF-alpha was quantified after varying the concentration of a primary low dose of LPS prior to a challenge with a high, normally stimulatory dose of LPS . The kinetics of establishment of the hyporesponsive state and the effect of varying the bacterial serotype and genus of the challenge dose were determined . The specificity of the hyporesponsive state for LPS was examined . Our results demonstrate that complete hyporesponsiveness with no detectable production of TNF-alpha (< 30 pg/ml) was achieved after a primary dose > or = 10 ng/ml . Establishment of the hyporesponsive state took place within 6 hr . Induction of hyporesponsiveness was not dependent upon the serotype or genus of the challenge dose of LPS and was not specific for LPS . Complete hyporesponsiveness was induced after a primary dose (10 micrograms/ml) of the Gram-positive bacterium Corynebacterium parvum (Cp) and was evident upon challenge with 100 micrograms/ml Cp . The data indicate that the mechanisms by which LPS and Cp induce hyporesponsiveness are not identical in that a primary dose of LPS (10 ng/ml) induced only partial hyporesponsiveness upon challenge with Cp (100 micrograms/ml) . These studies improve our understanding of Kupffer cell function. J Clin Microbiol, 1994 Mar, 32(3), 740 - 5 Rapid identification of mycolic acid patterns of mycobacteria by high-performance liquid chromatography using pattern recognition software and a Mycobacterium library; Glickman SE et al.; Current methods for identifying mycobacteria by high-performance liquid chromatography (HPLC) require a visual assessment of the generated chromatographic data, which often involves time-consuming hand calculations and the use of flow charts . Our laboratory has developed a personal computer-based file containing patterns of mycolic acids detected in 45 species of Mycobacterium, including both slowly and rapidly growing species, as well as Tsukamurella paurometabolum and members of the genera Corynebacterium, Nocardia, Rhodococcus, and Gordona . The library was designed to be used in conjunction with a commercially available pattern recognition software package, Pirouette (Infometrix, Seattle, Wash.) . Pirouette uses the K-nearest neighbor algorithm, a similarity-based classification method, to categorize unknown samples on the basis of their multivariate proximities to samples of a preassigned category . Multivariate proximity is calculated from peak height data, while peak heights are named by retention time matching . The system was tested for accuracy by using 24 species of Mycobacterium . Of the 1,333 strains evaluated, > or = 97% were correctly identified . Identification of M . tuberculosis (n = 649) was 99.85% accurate, and identification of the M . avium complex (n = 211) was > or = 98% accurate; > or = 95% of strains of both double-cluster and single-cluster M . gordonae (n = 47) were correctly identified . This system provides a rapid, highly reliable assessment of HPLC-generated chromatographic data for the identification of mycobacteria. J Steroid Biochem Mol Biol, 1994 Mar, 48(4), 409 - 18 Comparison of 16-androstene steroid concentrations in sterile apocrine sweat and axillary secretions: interconversions of 16-androstenes by the axillary microflora--a mechanism for axillary odour production in man? Gower DB, Holland KT, Mallet AI, Rennie PJ, Watkins WJ. The concentrations of five 16-androstene steroids were determined, by a GC-MS method, in freshly-produced apocrine sweat (adrenaline-induced), in 8 men and 2 women . The ranges of concentrations (nmol/microliter) in apocrine sweat were: 5 alpha-androst-16-en-3-one (5 alpha-A), 0.1-2.0 and 4,16-androstadien-3-one (androstadienone), 0-1.9, 5,16-Androstadien-3 beta-ol (androstadienol) was also found in 5 of the subjects (range 0.05-1.05) . 5 alpha-Androst-16-en-3 alpha- or 3 beta-ols {3 alpha (beta)-androstenols} were only found in small amounts (< 0.1 nmol/microliters) in a few subjects . In the second study, prior to apocrine sweat collection (adrenaline injection), the axillary skin of 6 of the male subjects was washed with diethyl ether on an adjacent site of the axillary vault . The concentrations of 16-androstenes were compared in the ethereal extracts and apocrine sweat . The former contained detectable levels (pmol/cm2) of androstadienone (17.9 +/- 2.4), 3 alpha-androstenol (6.9 +/- 3.7), 3 beta-androstenol (1.8 +/- 1.0) and androstadienol (1.9 +/- 0.5) (means +/- SEM) in all 6 subjects . All but 1 subject also had 5 alpha-androstenone, the mean value for the others being 2.5 +/- 0.6 . The axillary skin levels of 3 alpha- and 3 beta-androstenols, androstadienol and, in 3 subjects, androstadienone exceeded those in the apocrine sweat obtained from the same subjects, whereas levels of 5 alpha-androstenone in the skin extracts were all lower than in apocrine sweat samples, when related to the corresponding areas of skin sampled . The metabolism of 16-androstenes was studied in vitro in the presence of two aerobic coryneform bacteria, previously shown to metabolize testosterone as well as being capable of producing odour from extracts of axillary sweat in an odour-generation test . Although both coryneforms caused complex metabolic reactions and were capable of oxidation or reduction at C-3 and C-4, the overall direction favoured reduction . For example, large quantities of the more odorous 5 alpha-androstenone and 3 alpha-androstenol were formed from androstadienol and androstadienone . In contrast, strains of corynebacteria, unable to produce odour and incapable of metabolizing testosterone, were also unable to metabolize 16-androstenes.(ABSTRACT TRUNCATED AT 400 WORDS) J Clin Microbiol, 1994 Mar, 32(3), 854 - 5 Alpha-mannosidase: a rapid test for identification of Arcanobacterium haemolyticum; Carlson P et al.; A 4-h alpha-mannosidase test for identification of Arcanobacterium haemolyticum strains (n = 139) and differentiation of A . haemolyticum from Actinomyces pyogenes strains (n = 30) and other gram-positive rods was evaluated . Practically all A . haemolyticum strains (138 of 139) and the Listeria monocytogenes type strain were alpha-mannosidase positive, while all A . pyogenes strains and Corynebacterium (n = .25) strains as well as the Rhodococcus equi and Erysipelothrix rhusiopathiae type strains were negative . The rapid alpha-mannosidase test, in conjunction with a Gram stain and catalase and reverse CAMP tests, was useful in identification of A . haemolyticum and in differentiation of A . haemolyticum from A . pyogenes and Corynebacterium spp. J Leukoc Biol, 1994 Mar, 55(3), 362 - 70 LGL-1: a potential triggering molecule on murine NK cells; Mason LH et al.; Natural killer (NK) cells mediate non-major histocompatibility complex-restricted lysis of tumor cells, lymphokine-activated killing (LAK), antibody-dependent cellular cytotoxicity (ADCC), and reverse ADCC (RADCC) . LGL-1+ cells identify a major subset (50%) of murine NK cells . Here we demonstrate that monoclonal antibodies (mAbs) to LGL-1 consistently induce interleukin-2-cultured, and Corynebacterium parvum (in vivo)-activated NK cells to induce RADCC . LGL-1 triggering of activated NK cells coincides with enhanced LGL-1 expression . Testing of murine mAbs to epitopes of CD2 only appears to augment RADCC induced by mAb NK-1.1 on fresh NK cells . Immunoprecipitation of the LGL-1 antigen reveals a highly disulfide-linked 40-kDa homodimer subunit that is N-glycosylated . Therefore, LGL-1 may be similar to other recently characterized NK-associated antigens such as NK-1.1, Ly-49, and NKR-PI . We conclude that although LGL-1 is expressed on "resting" NK cells, enhanced surface expression following activation is usually required for it to act as a signaling molecule. Environ Res, 1994 Feb, 64(2), 181 - 91 Mesothelial cell injury caused by pleural leukocytes from rats treated with intratracheal instillation of crocidolite asbestos or Corynebacterium parvum; Li XY et al.; The pleural and peritoneal mesothelium is a major target in asbestos exposure where mesothelial cell proliferation, exfoliation, and neoplasia have been reported in workers and experimental animals . The objective of this study was to determine the role of pleural leukocytes in mesothelial cell damage caused by asbestos exposure . We therefore investigated detachment and lysis injury to mesothelial cells in vitro induced by leukocytes lavaged from the pleural space of rats exposed, by intratracheal instillation, to crocidolite asbestos . Our studies revealed that normal pleural leukocytes were composed of macrophages, lymphocytes, eosinophils, and mast cells . This population showed a small but significant recruitment of mast cells and eosinophils 3 to 14 days after instillation of crocidolite asbestos; there were also modestly increased levels of macrophages present 30 days after low doses of asbestos . One day after intratracheal instillation of 5 mg crocidolite asbestos, the pleural leukocytes caused detachment injury to mesothelial cells in vitro . This injury was markedly reduced 3 days after asbestos exposure and was undetectable by Day 14 . One month after instillation of asbestos, despite doses of asbestos from 0.5 to 5 mg, pleural leukocytes showed no ability to injure mesothelial cells in vitro . In a parallel study, pleural inflammation was induced by intratracheal instillation of heat-inactivated Corynebacterium parvum . Transient mesothelial cell-detaching injury was again expressed by pleural leukocytes 1 day after C . parvum instillation . This was likely related to an increase in the percentage of neutrophils present on this day . These results show that a single administration of crocidolite asbestos, intratracheally, leads to transient activation of pleural leukocytes in terms of the ability of these cells to detach mesothelial cells from matrix . This finding implies that mesothelial cell proliferation and exfoliation found in individuals exposed to asbestos may result from persistent stimulation of pleural leukocytes caused by the continuous presence of asbestos in the lung. Mol Microbiol, 1994 Feb, 11(4), 739 - 46 Isolation and characterization of IS31831, a transposable element from Corynebacterium glutamicum; Vertes AA et al.; A transposable element from a coryneform bacterium, Corynebacterium glutamicum