Exp Anim, 2002 Jan, 51(1), 99 - 102 Detection of Corynebacterium kutscheri from the oral cavity of rats; Amao H et al.; A simple and useful method for the detection of C . kutscheri from the oral cavity of living rats was devised . In 10 sacrificed rats from two naturally and subclinically infected conventional colonies, 10(4.28) or 10(3.84) CFU/ml C . kutscheri were isolated from upper incisor swab extractions, while 10(1.38) or 10(1.58) and < 10 or 10(1.56) CFU/ml from the upper soft palate and pharynx, respectively . In another survey with 26 living animals, which were reared on the same rack, organisms were detected from the upper incisor and gingival swabs in 15 of 26 rats (57.7%) . The results were reproducible at a second survey 10 days later . No organisms were isolated from any sites of the orally negative rats . These results indicated that culture of swab specimens from the upper incisors and gingivae of incisors is useful for the detection of C . kutscheri infection in living rats. Clin Microbiol Infect, 1996 Feb, 2(3), 209 - 213 Disk diffusion, agar dilution and the E-test for susceptibility testing of Corynebacterium jeikeium; Pennekamp A et al.; OBJECTIVE: The susceptibilities to penicillin, tetracycline, erythromycin, gentamicin, vancomycin and teicoplanin of 58 strains of Corynebacterium jeikeium were assessed by disk diffusion and agar dilution reference methods . METHODS: Zone sizes and minimal inhibitory concentrations (MIC) by agar dilution were interpreted using the ranges in the NCCLS tables for organisms other than Haemophilus, Neisseria gonorrhoeae, and Streptococcus pneumoniae . RESULTS: By agar dilution, 14%, 88%, 17% and 26% of the 58 isolates were susceptible to penicillin, tetracycline, erythromycin, and gentamicin, respectively . Using the breakpoints for Listeria monocytogenes, all strains showed concordant results for penicillin by disk diffusion . Discrepancies in the interpretative categories by disk diffusion were found in four cases (two very major and two minor) for tetracycline, in nine (two very major, two major, and five minor) for erythromycin, and in 1 case (very major) for gentamicin . All 58 strains were susceptible to vancomycin and teicoplanin by agar dilution and disk diffusion . The overall agreement of interpretative disk diffusion for all six antibiotics was 95.9% . In addition, all strains were susceptible to both glycopeptides by E-test . However, for vancomycin the MIC results in 58.6% were two log2 dilutions and in 1.7% more than two log2 dilutions higher by E-test than by agar dilution, whereas for teicoplanin agreement within one log2 dilution was 100% . CONCLUSIONS: Further evaluation of methodologies of disk diffusion is required to obtain a better agreement for erythromycin and tetracycline . The criteria of the NCCLS for interpretation of disk diffusion are adequate for susceptibility testing of C . jeikeium to penicillin, gentamicin, vancomycin and teicoplanin. Clin Microbiol Infect, 1997 Feb, 3(6), 634 - 639 Clinical significance of Corynebacterium striatum isolated from human samples; Martinez-Martinez L et al.; OBJECTIVE: To evaluate the clinical significance of and describe factors associated with Corynebacterium striatum infection . METHODS: A retrospective chart review was performed of the C . striatum isolated in a university hospital from January 1991 to July 1995 . C . striatum was identified using conventional methods, the API CORYNE system and cellular fatty acid profiles . RESULTS: In the study period, C . striatum was isolated from clinical samples in 127 patients . In 49 patients, data from clinical charts were considered insufficient for evaluation . In 26 cases, the microorganism was considered to be the etiologic agent of an infectious process . In the remaining 52 patients, the organism was considered to be a colonizer . Before the infection all the patients had been hospitalized for some underlying condition, and 22 (85%) of them had received antibiotics previously . Six patients died . In two of them, death was a consequence of their underlying disease and in the remaining four, death was related to the C . striatum infection . CONCLUSIONS: C . striatum, a microorganism traditionally considered to be an avirulent member of the normal human nasopharyngeal and skin flora, may opportunistically cause infections in hospitalized patients with underlying diseases and previous antibiotic treatments. Clin Microbiol Infect, 1997, 3(5), 555 - 558 Efficacy of a selective and differential medium for isolating Corynebacterium urealyticum from urine specimens; Garcia-Bravo M et al.; OBJECTIVE: To describe a new selective and differential medium for improving the isolation of Corynebacterium urealyticum from urine . METHODS: A total of 370 urine specimens, 270 from patients with underlying urologic disease and 100 from to a control group without renal pathology, were cultured on standard sheep blood agar medium and on a new selective medium for Corynebacterium urealyticum . RESULTS: The use of this selective medium resulted in a nine-fold increase in the prevalence of bacteriuria caused by C . urealyticum (12.4% versus 1.4%) . Furthermore, the frequency of patients with symptomatic bacteriuria detected with the selective medium (3.2%) was higher than with the non-selective medium (1.4%) . CONCLUSIONS: We recommend the use of this selective medium for the isolation of C . urealyticum in patients with underlying urologic disease or with chronic urinary symptoms. J Dairy Sci, 2002 Jan, 85(1), 112 - 21 The effect of selective dry cow treatment on new intramammary infections; Berry EA et al.; Dry cow therapy, or antibiotic treatment at end of lactation, is used to eliminate intramammary infections and prevent new infections during the dry period . It is one part of a total management system recommended in controlling intramammary infections in the dairy cow . Public health concerns advise prudent use of antibiotics, as their use may promote bacterial antibiotic resistance and leave antibiotic residues in the food chain . The effects of dry cow treatment and no treatment were compared, on new intramammary infections and clinical mastitis within two low cell count herds and two herds undergoing conversion to organic farming . The results will inform those restricting their use of dry cow therapy on the additional risk of new intramammary infection and aid in development of alternative management strategies . No cases of clinical mastitis in the dry period were observed in treated cows, whereas in the untreated groups a significant number were observed . Significantly more new infections at calving were found in the untreated group in all herds . In those quarters where infections were first detected at calving, the incidence of clinical mastitis was significantly greater in the untreated group in all herds . Clinical mastitis detection was significantly lower in organic herds . Untreated quarters infected at drying with Corynebacterium spp . or coagulase-negative staphylococci were found to have an increased risk of new infection by Streptococcus uberis or coliform bacteria . It can be concluded that dry cow therapy continues to lower significantly the rate of new dry period intramammary infection in herds with elevated somatic cell counts and a high prevalence of infection. Diagn Microbiol Infect Dis, 2002 Feb, 42(2), 119 - 22 In vitro evaluation of AZD2563, a new oxazolidinone, tested against unusual gram-positive species; Jones RN et al.; The recently introduced oxazolidinone, linezolid, has a spectrum and potency directed against Gram-positive organisms, including antimicrobial-resistant isolates . The newest agent in this class, AZD2563 was tested against uncommonly isolated Gram-positive species to establish the breadth of its spectrum . By reference broth microdilution methods, 120 strains were tested (48 Corynebacterium spp., 10 species; 27 Listeria spp., 2 species; 11 Micrococcus spp., 2 species; 23 Bacillus spp., 3 species; 6 Stomatococcus mucilaginosus and one strain each of 5 other species) against AZD2563 and compared to eight other agents . The AZD2563/linezolid MIC(50;) % inhibited at < or =4 microg/mL were: for corynebacteria (0.25/0.25 microg/mL; 100/100%), Listeria spp . (2/2 microg/mL; 100/100%), Micrococcus spp . (1/1 microg/mL; 100/100%), Bacillus spp . (0.5/1 microg/mL; 100/100%), and S mucilaginosus (0.5/1 microg/mL; 100/100%) . Using the MIC(90) values, AZD2563 was slightly more potent than linezolid (two-fold) . Only four genus- species groups had AZD2563 MICs of strains at 2 microg/mL (Aerococcus, Leuconostoc, Listeria, Rhodococcus), all other isolates were inhibited by < or = 1 microg/mL . The AZD2563 potency and spectrum versus these rarer species was at least equal to linezolid, including some strains resistant to penicillins, macrolides-lincosamides, and fluoroquinolones. Tidsskr Nor Laegeforen, 2002 Jan 10, 122(1), 62 - 3 {Axel Ström--pioneer of social medicine and administrator}; Sundby P; Dr Axel Strom (1901-85), professor in the University of Oslo from 1940 to 1970, was a leader in Norwegian medicine in the latter half of the 20th century . He qualified in 1926 and in 1936 gained a doctorate with a dissertation on the toxin production of the Corynebacterium diphtheriae . His first appointment as a professor was in hygiene . In 1951 he moved on to public health, a field that he pioneered in Norway and the other Scandinavian countries . As a professor during the German occupation of Norway in the Second World War, he joined the university's resistance against the Nazi authorities' attempts at taking control . When the war was over he became deeply involved in research on the impact of war on health . At a time when the study of the impact of lifestyle factors was still in its infancy, he suggested that the war-induced reduction in dietary fat consumption might be the cause of observed lower cardiovascular mortality . Of more practical importance were the studies he initiated of the mainly psychological late-onset effects of traumas suffered by prisoners in German camps, seamen, soldiers and other exposed groups . In this area, too, he was an early explorer, of what has come to be known as post-traumatic stress disorder . His efforts led to improved war pension entitlements for the victims . Over the years, exposed groups became his major professional interest as a public health specialist . In his academic work, dr Strom also pioneered medical ethics, care for the elderly, legislation on abortion, and the rapidly expanding field of the medical basis for social security benefits . As a practising physician he was in the vanguard of occupational medicine and other kinds of preventive medicine . What brought him most recognition was, however, his leading role over many years in the Norwegian Medical Association and in the University of Oslo . He served as chairman of the Junior Hospital Doctors Association, president of the Norwegian Medical Association and chairman of the Federation of Norwegian Professional Associations . He was elected dean of the Faculty of Medicine and vice-rector of the University of Oslo, in addition to a host of other expert assignments and official roles: He was renowned for his hard work and exerted great influence in many quarters. Arch Dermatol, 1965 Jul, 92(1), 88 - 90 Levels of antibody to Staphylococcus epidermidis in patients with acne vulgaris; Puhvel SM et al.; Sera from 23 patients with acne vulgaris of varying degrees and sera from 15 patients with skin diseases other than acne were tested for antibody levels to Staphylococcus epidermidis by use of bacterial agglutination and agar-gel immunodiffusion . Antibody levels to S epidermidis varied from 0 to 1:160 in both the patients with acne and in the control groups . There was no correlation between the antibody level to S epidermidis and the degree of acne . In a previous investigation it was found that antibody levels to Corynebacterium acnes are significantly increased in serum from patients with papulopustular and cystic acne . Both S epidermidis and C acnes can frequently be isolated from lesions in acne . The fact that antibody levels are increased to C acnes but not to S epidermidis may indicate that the mere presence of an organism in the acne lesion is not sufficient stimulation for antibody formation . The increase of antibody to C acnes which was demonstrated previously in patients with severe acne vulgaris may therefore reflect a direct involvement of the organism in the acne process. J Appl Microbiol, 2002, 92(2), 215 - 20 The amino acid requirements of Corynebacterium diphtheriae PW 8 substrain CN 2000; Nagarkar PP et al.; AIMS: To determine the amino acid requirement and utilization pattern of Corynebacterium diphtheriae during growth and toxin production . METHODS AND RESULTS: Comparing across different batches of beef-based media, the growth and toxin yield were correlated significantly with nine of the amino acids . The amino acid utilization pattern during growth of C . diphtheriae further showed that only four of the nine amino acids, namely cystine, histidine, aspartate and methionine, were critical for growth of the vaccine strain . Further investigations using synthetic media with combinations of amino acid supplements demonstrated that among the four, cystine was the most growth limiting . CONCLUSIONS: Only certain amino acids are critical for growth and toxin production by C . diphtheriae, cystine being the single most important . SIGNIFICANCE AND IMAPCT OF THE STUDY: Owing to the potential threat from Bovine Spongiform Encephalopathy (BSE), a need is recognized by vaccine manufacturers to substitute beef-based production media . An understanding of the specific amino acid requirements would help to develop and optimize alternative production media. J Bacteriol, 2002 Mar, 184(5), 1277 - 86 Corynebacterium glutamicum utilizes both transsulfuration and direct sulfhydrylation pathways for methionine biosynthesis; Hwang BJ et al.; A direct sulfhydrylation pathway for methionine biosynthesis in Corynebacterium glutamicum was found . The pathway was catalyzed by metY encoding O-acetylhomoserine sulfhydrylase . The gene metY, located immediately upstream of metA, was found to encode a protein of 437 amino acids with a deduced molecular mass of 46,751 Da . In accordance with DNA and protein sequence data, the introduction of metY into C . glutamicum resulted in the accumulation of a 47-kDa protein in the cells and a 30-fold increase in O-acetylhomoserine sulfhydrylase activity, showing the efficient expression of the cloned gene . Although disruption of the metB gene, which encodes cystathionine gamma-synthase catalyzing the transsulfuration pathway of methionine biosynthesis, or the metY gene was not enough to lead to methionine auxotrophy, an additional mutation in the metY or the metB gene resulted in methionine auxotrophy . The growth pattern of the metY mutant strain was identical to that of the metB mutant strain, suggesting that both methionine biosynthetic pathways function equally well . In addition, an Escherichia coli metB mutant could be complemented by transformation of the strain with a DNA fragment carrying corynebacterial metY and metA genes . These data clearly show that C . glutamicum utilizes both transsulfuration and direct sulfhydrylation pathways for methionine biosynthesis . Although metY and metA are in close proximity to one another, separated by 143 bp on the chromosome, deletion analysis suggests that they are expressed independently . As with metA, methionine could also repress the expression of metY . The repression was also observed with metB, but the degree of repression was more severe with metY, which shows almost complete repression at 0.5 mM methionine in minimal medium . The data suggest a physiologically distinctive role of the direct sulfhydrylation pathway in C . glutamicum. Vet Microbiol, 2002 Mar 1, 85(2), 133 - 44 Phenotypic and genetic characterisation of bacteriocin-producing strains of Staphylococcus aureus involved in bovine mastitis; dos Santos Nascimento J et al.; Fifty strains of Staphylococcus spp . isolated from bovine mastitis cases in several herds from different Argentinian provinces were screened for antimicrobial substances . Twelve strains exhibited a high antagonistic activity against the indicator strain (Corynebacterium fimi) and were chosen for further characterisation . The antimicrobial substances were sensitive to proteolytic enzymes suggesting that they might be bacteriocins (Bac) . These strains were identified as S . aureus by the amplification of the femA gene . Plasmid profile analysis of these strains revealed the presence of at least one plasmid . Eleven strains carried a plasmid with a size similar to that of pRJ6 (8.0kb), which encodes aureocin A70, a bacteriocin produced by the Brazilian S . aureus strain A70 isolated from commercial milk . The other strain harboured a much larger plasmid . PCR experiments, using specific primers for amplification of the bacteriocin operon found in pRJ6, showed that all strains had the expected 525bp amplicon, suggesting that the bacteriocin produced may be related to aureocin A70 . The genomic DNA of all Bac(+) strains was then analysed by pulsed-field gel electrophoresis (PFGE) in order to investigate clonal relationships amongst strains . Based on the results of PFGE experiments, 10 out of the 12 Bac(+) strains belonged to the same clone . The remaining two strains are possibly related to the prevalent clone . The aureocin A70 producer-strain belonged to a distinct clone. Int J Syst Evol Microbiol, 2002 Jan, 52(Pt 1), 91 - 100 Identification of coryneform bacteria and related taxa by Fourier-transform infrared (FT-IR) spectroscopy; Oberreuter H et al.; An extensive Fourier-transform infrared (FT-IR) spectroscopy database for the identification of bacteria from the two suborders Micrococcineae and Corynebacterineae (Actinomycetales, Actinobacteria) as well as other morphologically similar genera was established . The database consists of averaged IR spectra from 730 reference strains, covering 220 different species out of 46 genera . A total of 192 species are represented by type strains . The identity of 352 reference strains was determined by comparative 16S rDNA sequence analysis and, if necessary, strains were reclassified accordingly . FT-IR frequency ranges, weights and reproducibility levels were optimized for this section of high-G+C gram-positive bacteria . In an internal validation, 98.1% of 208 strains were correctly identified at the species level . A simulated external validation which was carried out using 544 strains from 54 species out of 16 genera resulted in a correct identification of 87.3% at the species level and 95.4% at the genus level . The performance of this identification system is well within the range of those having been reported in the literature for the identification of coryneform bacteria by phenotypical methods . Coryneform and related taxa display a certain degree of overlapping distribution of different taxonomical markers, leading to a limited differentiation capacity of non-genotypical identification methods in general . However, easy handling, rapid identification within 25 h starting from a single colony, a satisfactory differentiation capacity and low cost, render FT-IR technology clearly superior over other routine methods for the identification of coryneform bacteria and related taxa. Appl Microbiol Biotechnol, 2002 Jan, 58(1), 89 - 96 Expression of genes of lipid synthesis and altered lipid composition modulates L-glutamate efflux of Corynebacterium glutamicum; Nampoothiri KM et al.; L-Glutamate is made with Corynebacterium glutamicum on a scale of more than 106 tons/year . Nevertheless, formation of this amino acid is enigmatic and there is very limited molecular information available to unravel the apparently complex conditions leading to L-glutamate efflux . Here, we report the isolation and overexpression of the genes involved in lipid synthesis: acp, fadD 15, cma, cls, pgsA2, cdsA, gpsA, and plsC, and the inactivation of cma and cls . In addition, the consequences for phospholipid content, temperature sensitivity, as well as detergent-independent and detergent-dependent L-glutamate efflux were quantified . An in part strong alteration of the phospholipid composition was achieved; for instance, overexpression offadD15 encoding an acyl-CoA ligase resulted in an increase of phosphatidyl inositol from 12.6 to 30.2% . All strains, except that overexpressing acp (acyl carrier protein), exhibited increased temperature sensitivity, with the strongest sensitivity present upon cls (cardiolipin synthetase) inactivation . As a consequence of the genetically modified lipid synthesis, L-glutamate efflux changed quite dramatically; for instance, overexpression of plsC (acylglycerolacyl transferase) resulted in a detergent-triggered increase of L-glutamate accumulation from 92 mM to 108 mM, whereas acp overexpression reduced the accumulation to 24 mM . With some of the overexpressed genes, substantial L-glutamate excretion even without detergent addition was obtained when the fermentation temperature was elevated . These data show that the chemical and physical properties of the cytoplasmic membrane are altered and suggest that this is a necessary precondition to achieve L-glutamate efflux. Electrophoresis, 2001 Dec, 22(20), 4404 - 22 A high-resolution reference map for cytoplasmic and membrane-associated proteins of Corynebacterium glutamicum; Schaffer S et al.; We present a high-resolution reference map for soluble proteins obtained from Corynebacterium glutamicum cells grown in glucose minimal medium . The analysis window covers the pl range from 4-6 and the molecular mass range from 5-100 kDa . Using overlapping narrow immobilized pH gradients for isoelectric focusing, 970 protein spots were detected after second-dimensional separation on SDS-polyacrylamide gels and colloidal Coomassie-staining . By tryptic peptide mass fingerprinting 169 protein spots were identified, representing 152 different proteins including many enzymes involved in central metabolism (18), amino acid biosynthesis (24) and nucleotide biosynthesis (11) . Thirty-five of the identified proteins have no known function . A comparison of the observed and the expected physicochemical properties of the identified proteins indicated that nine proteins were covalently modified, since variants with apparently identical molecular mass, but differing pl were detected . The N-termini of eight proteins were determined by post-source decay (PSD) analysis of selected peptides . In addition to the soluble proteins, a map of the membrane-bound proteins within the pl range 4-7 is presented, which contains 660 protein spots, 22 of which were identified, representing 13 different proteins. FEMS Immunol Med Microbiol, 2002 Jan 14, 32(2), 141 - 7 Priming and activation of mouse macrophages by trehalose 6,6'-dicorynomycolate vesicles from Corynebacterium glutamicum; Chami M et al.; Vesicles consisting of pure trehalose dicorynomycolate (TDCM), the corynebacterial analog of the most studied mycobacterial glycolipid 'cord factor', were isolated from Corynebacterium glutamicum cells by mild detergent treatment; these induced in vivo a macrophage priming similar to that obtained with mycobacterial-derived trehalose dimycolate . In vitro, both TDCM and bacterial lipopolysaccharide (LPS) induced in macrophages the production of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), endotoxin tolerance, and were primed for an enhanced secondary NO response to LPS . Interferon-gamma pretreatment did not influence the LPS-induced TNF-alpha response, but considerably increased the TDCM-induced response. Adv Biochem Eng Biotechnol, 2001, 73, 9 - 29 Metabolic engineering for L-lysine production by Corynebacterium glutamicum; de Graaf AA et al.; Corynebacterium glutamicum has been used since several decades for the large-scale production of amino acids, esp . L-glutamate and L-lysine . After initial successes of random mutagenesis and screening approaches, further strain improvements now require a much more rational design, i.e . metabolic engineering . Not only recombinant DNA technology but also mathematical modelling of metabolism as well as metabolic flux analysis represent important metabolic engineering tools . This review covers as state-of-the-art examples of these techniques the genetic engineering of the L-lysine biosynthetic pathway resulting in a vectorless strain with significantly increased dihydrodipicolinate synthase activity, and the detailed metabolic flux analysis by 13C isotopomer labelling strategies of the anaplerotic enzyme activities in C . glutamicum resulting in the identification of gluconeogenic phosphoenolpyruvate carboxykinase as a limiting enzyme. J Dairy Sci, 2001 Dec, 84(12), 2649 - 63 Cow- and quarter-level risk factors for Streptococcus uberis and Staphylococcus aureus mastitis; Zadoks RN et al.; This study was designed to identify risk factors for intramammary infections with Streptococcus uberis and Staphylococcus aureus under field conditions . An 18-mo survey with sampling of all quarters of all lactating cows at 3-wk intervals was carried out in three Dutch dairy herds with medium bulk milk somatic cell count (200,000 to 300,000 cells/ml) . Quarter milk samples were used for bacteriology and somatic cell counting . Data on parity, lactation stage, and bovine herpesvirus 4-serology were recorded for each animal . During the last year of the study, body condition score, and teat-end callosity scores were recorded at 3-wk intervals . A total of 93 new infections with Strep . uberis were detected in 22,665 observations on quarters at risk for Strep . uberis infection, and 100 new infections with Staph . aureus were detected in 22,593 observations on quarters at risk for Staph . aureus infection . Multivariable Poisson regression analysis with clustering at herd and cow level was used to identify risk factors for infection . Rate of infection with Strep . uberis was lower in first- and second-parity cows than in older cows, and depended on stage of lactation in one herd . Quarters that were infected with Arcanobacterium pyogenes or enterococci, quarters that had recovered from Strep . uberis- or Staph . aureus-infection in the past, and quarters that were exposed to another Strep . uberis infected quarter in the same cow had a higher rate of Strep . uberis infection . Teat-end callosity and infection with coagulase-negative staphylococci or corynebacteria were not significant as risk factors . Rate of Staph . aureus infection was higher in bovine herpesvirus 4-seropositive cows, in right quarters, in quarters that had recovered from Staph . aureus or Strep . uberis infection, in quarters exposed to other Staph . aureus infected quarters in the same cow, and in quarters with extremely callused teat ends . Infection with coagulase-negative staphylococci was not significant as a risk factor . The effect of infection with corynebacteria on rate of infection with Staph . aureus depended on herd, stage of lactation, and teat-end roughness . Herd level prevalence of Strep . uberis or Staph . aureus, and low quarter milk somatic cell count were not associated with an increased rate of infection for Strep . uberis or Staph . aureus. Biochemistry, 2002 Jan 29, 41(4), 1381 - 9 Crystal structure of the di-iron/radical protein of ribonucleotide reductase from Corynebacterium ammoniagenes; Hogbom M et al.; Ribonucleotide reductase (RNR) is the enzyme performing de novo production of the four deoxyribonucleotides needed for DNA synthesis . All mammals as well as some prokaryotes express the class I enzyme which is an alpha(2)beta(2) protein . The smaller of the homodimers, denoted R2, contains a di-iron carboxylate site which, upon reaction with molecular oxygen, generates a stable tyrosyl radical needed for catalysis . The three-dimensional structure of the oxidized class Ib RNR R2 from Corynebacterium ammoniagenes has been determined at 1.85 A resolution and refined to an R-value of 15.8% (R(free) = 21.3%) . In addition, structures of both the reduced iron-containing, and manganese-substituted protein have been solved . The C . ammoniagenes R2 has been proposed to be manganese-dependent . The present structure provides evidence that manganese is not oxidized by the protein, in agreement with recent biochemical data, and that no obvious structural abnormalities are seen in the oxidized and reduced iron-containing forms, giving further support that the protein is indeed an iron-dependent RNR R2 . The di-manganese structure also provides an explanation for the magnetic properties of this site . The structure of the oxidized C . ammoniagenes R2 also reveals an additional water molecule bridging the radical and the iron site, which has not previously been seen in any other R2 structure and which might have important mechanistic implications. Laryngoscope, 2001 Nov, 111(11 Pt 1), 2054 - 9 Microbiology of normal external auditory canal; Stroman DW et al.; OBJECTIVES: To isolate and characterize bacteria and fungi from the healthy ear and to obtain susceptibility profiles on each bacterial isolate . STUDY DESIGN: Prospective . METHODS: Specimens were collected from the external canals and cerumen of healthy subjects . Species-level identification was obtained by combining phenotypic and genotypic data . End-point minimal inhibitory concentration testing was performed using National Committee for Clinical Laboratory Standards recommended methods . RESULTS: One hundred sixty-four subjects were cultured . Seventeen canal and 16 cerumen specimens showed no growth . One hundred forty-eight cerumen specimens yielded 314 organisms, including 23 fungi . One hundred forty-seven canal specimens yielded 310 organisms, including 7 fungi . Of 291 bacteria isolated from cerumen, 99% were Gram-positive . Of 302 bacteria isolated from the canal, 96% were Gram-positive . Staphylococci were 63% of both the cerumen bacteria and the canal bacteria . Coryneforms represented 22% of the bacteria in cerumen and 19% in the canal . Turicellaotitidis was the primary coryneform isolated from both the canal and the cerumen . Streptococci-like bacteria were 10% from the cerumen, 7% from the canal . In both cerumen and canal, Alloiococcusotitis was more than 95% of the streptococci-like bacteria . Fifteen gram-negative organisms were isolated from the canal and cerumen, including four Pseudomonas aeruginosa strains . The percentages of Staphylococcus epidermidis isolates that had high-level resistance (> or =8 microg/mL) were as follows: to neomycin, 28% from cerumen and 11% from the canal; to oxacillin, 28% from cerumen and 25% from the canal; and to ofloxacin, 15% from cerumen and 19% from the canal . CONCLUSIONS: Turcellaotitidis and A . otitidis were present with a much higher frequency than previously described, lending evidence that they be considered normal otic flora . Corynebacterium auris, previously reported only in children, was isolated from normal adults. Appl Microbiol Biotechnol, 2001 Dec, 57(5-6), 667 - 73 Metabolic redirection of carbon flow toward isoleucine by expressing a catabolic threonine dehydratase in a threonine-overproducing Corynebacterium glutamicum; Guillouet S et al.; Carbon destined for lysine synthesis in Corynebacterium glutamicum ATCC 21799 can be diverted toward threonine by overexpression of genes encoding a feedback-insensitive homoserine dehydrogenase (hom(dr)) and homoserine kinase (thrB) . We studied the effects of introducing two different threonine dehydratase genes into this threonine-producing system to gauge their effects on isoleucine production . Co-expression of hom(dr), thrB, and ilvA, which encodes a native threonine dehydratase, caused isoleucine to accumulate to a final concentration of 2.2+/-0.2 g l(-1), five-fold more than accumulates in the wild-type strain, and approximately twice as much as accumulates in the strain expressing only hom(dr) and thrB . Comparing these data with previous results, we found that overexpression of the three genes, hom(dr), thrB, and ilvA, in C . glutamicum ATCC 21799 is no better in terms of isoleucine production than the expression of a single gene, tdcB, encoding a catabolic threonine dehydratase from Escherichia coli . Co-expression of hom(dr), thrB, and tdcB, however, caused the concentration of isoleucine to increase 20-fold compared to the wild-type strain, about four times more than the corresponding ilvA-expressing strain . In this system, the apparent yield of isoleucine production was multiplied by a factor of two {2.1 mmol (g dry cell weight)(-1)} . While the balance of excreted metabolites showed that the carbon flow in this strain was completely redirected from the lysine pathway into the isoleucine pathway, it also showed that more pyruvate was diverted into amino acid synthesis. J Antibiot (Tokyo), 2001 Oct, 54(10), 827 - 30 YUA001, a novel aldose reductase inhibitor isolated from alkalophilic Corynebacterium sp . YUA25 . II . Chemical modification and biological activity; Sun WS et al.; A series of novel N-substituted tyramine (2-p-hydroxyphenylethylamine) derivatives (1 to approximately 11) were synthesized and evaluated for their inhibitory activity against pig kidney aldose reductase (EC 1, 1, 1, 21) . Of these compounds, N-2-p-hydroxyphenylethyl maleamic acid (10) exhibits the strongest aldose reductase inhibitory activity, which is 22 times more potent than that of YUA001. J Clin Microbiol, 2002 Jan, 40(1), 80 - 3 Immunochromatographic strip test for rapid detection of diphtheria toxin: description and multicenter evaluation in areas of low and high prevalence of diphtheria; Engler KH et al.; An immunochromatographic strip (ICS) test was developed for the detection of diphtheria toxin by using an equine polyclonal antibody as the capture antibody and colloidal gold-labeled monoclonal antibodies specific for fragment A of the diphtheria toxin molecule as the detection antibody . The ICS test has been fully optimized for the detection of toxin from bacterial cultures; the limit of detection was approximately 0.5 ng of diphtheria toxin per ml within 10 min . In a comparative study with 915 pure clinical isolates of Corynebacterium spp., the results of the ICS test were in complete agreement with those of the conventional Elek test . The ICS test was also evaluated for its ability to detect toxigenicity from clinical specimens (throat swabs) in two field studies conducted within areas of the former USSR where diphtheria is epidemic . Eight hundred fifty throat swabs were examined by conventional culture and by use of directly inoculated broth cultures for the ICS test . The results showed 99% concordance (848 of 850 specimens), and the sensitivity and specificity of the ICS test were 98% (95% confidence interval, 91 to 99%) and 99% (95% confidence interval, 99 to 100%), respectively. Acta Otorrinolaringol Esp, 2001 Nov-Dec, 52(8), 690 - 6 {Analysis of the relationship between chronic pharyngitis and tonsillectomy with the microbiologic study of the cavum}; Lopez Gonzalez MA et al.; Chronic pharyngitis is an usual process with difficult treatment . It has been related to tonsillectomy . This study collected 224 patients suffering from chronic pharyngitis, 55 tonsillectomies and 169 without operation . Bacteriological culture was done from nasopharynx in the different seasons . Microorganisms more frequent were Staphylococcus aureus, Corynebacterium sp and Aspergillus . The time between the tonsillectomy and this study has been of more than 17 years . No relationship has been found between chronic pharyngitis and tonsillectomy, considering types of microorganisms, seasons, numbers of microorganisms isolated per culture, Gram stain, and age of the patients. Trop Anim Health Prod, 2001 Dec, 33(6), 511 - 9 Treatment trial of subclinical mastitis with the herb Persicaria senegalense (Polygonaceae); Abaineh D et al.; The possible remedial effect of Persicaria senegalense in bovine subclinical mastitis was studied by in vitro and in vivo antimicrobial tests, using crude extracts and the leaf in different forms . The in vitro test showed that isolates of Staphylococcus aureus, Candida albicans and Corynebacterium bovis from subclinical cases and an isolate of Pseudomonas aeruginosa from a clinical case of mastitis were all inhibited by the three crude extracts at 820 micrograms concentration . An in vivo trial feeding 1.5 kg of the cooked leaf per day for 5 days did not give a significant cure rate, whereas a second trial in which 0.77 kg of leaf powder, equivalent to 3 kg of wet leaf, was fed per day for 5 days resulted in an apparent cure rate of 92.8% (52.8% actual as there was a 40% spontaneous cure rate in the negative control group, in contrast to 80% (40% actual) in the positive control group treated with an intramammary antibiotic preparation . The difference in cure rate between the negative control group and the experimental group given 0.77 kg leaf powder was significant (p = 0.008). Rev Med Univ Navarra, 2001 Apr-Jun, 45(2), 9 - 13 {Infection associated with intravascular catheter in 1998 at the University Clinic of Navarra}; del Pozo JL et al.; PURPOSE: To study the intravascular catheter related infections (CRI) since January of 1998 to January of 1999 in our hospital . METHODS: We studied 540 catheter tips using a modified combination of the cuantitative method of Cleri and the semicuantitative method of Maki . The catheters were classified into two groups according to the presence or absence of criteria for CRI . RESULTS: 74.5% of the retired catheters because of infection suspice did not satisfied criteria for CRI . 44.7% of the patients with criteria suffered a catheter-related bacteriemia while just 1.7% of the patients without criteria suffered a bacteriemia . The most common isolated microorganisms were coagulase-negative Staphylococci, Corynebacterium species and S . aureus . DISCUSSION AND CONCLUSIONS: At least, 74.5% of the patients with a suspice of catheter related infection could undergo a non invasive diagnosis procedure that would have showed that the catheter was not infected. Biotechnol Bioeng, 2002 Jan 20, 77(2), 131 - 41 Optimization of L-phenylalanine production of Corynebacterium glutamicum under product feedback inhibition by elevated oxygen transfer rate; Shu CH et al.; Production feedback inhibition both on cell growth and on product formation of phenylalanine fermentation might be alleviated by elevated oxygen supply . Batch fermentations by a high phenylalanine producing strain Corynebacterium glutamicum CCRC 18335 at various initial phenylalanine concentrations (P(0)) ranging from 0 to 20 g/L and different oxygen transfer rate coefficients (K(L)a) ranging from 23 to 76 h(-1) were studied . The fermentation parameters with respect to P(0) were strongly dependent on K(L)a . Cell yield favored higher K(L)a and lower P(0) . Product yield with respect to varying phenylalanine concentration was evaluated by the relative oxygen availability (ROA) . The optimal ROA for phenylalanine formation was strongly dependent on the product concentration . While P(0) was low, the product inhibition was less significant and the maximum product yield occurred while ROA was at 0.5-0.6 . While P(0) was high, the product inhibition was significant and the maximum product yield occurred while ROA was at 0.8-0.9 . These results suggest that the product feedback inhibition of phenylalanine fermentation processes can be alleviated by a gradual increase in oxygen supply rate while the increasing product concentration is taken into account . The strategy is demonstrated in a fed-batch culture with elevated oxygen supply . The final phenylalanine concentration was 23.2 g/L, which was 45% better than that of the fed-batch fermentation without elevated oxygen supply . Likewise, the maximum productivity was improved by 42% at 0.37 g/(L x h) . J Antimicrob Chemother, 2002 Jan, 49(1), 165 - 8 Detection of transposon Tn5432-mediated macrolide-lincosamide-streptogramin B (MLSB) resistance in cutaneous propionibacteria from six European cities; Ross JI et al.; Forty-five cutaneous propionibacterial isolates from six European cities were found to be highly resistant to all macrolide-lincosamide-streptogramin B antibiotics, including the ketolide telithromycin . This contrasts with previously documented phenotypes associated with 23S rRNA mutations . Sequencing of the resistance determinant showed it to be erm(X) of corynebacterial origin located on the composite transposon Tn5432. FEMS Microbiol Lett, 2001 Dec 18, 205(2), 361 - 7 Nitrogen and carbon regulation of glutamine synthetase and glutamate synthase in Corynebacterium glutamicum ATCC 13032; Schulz AA et al.; The effect of nitrogen and carbon status on the regulation of glutamine synthetase (GS) and glutamate synthase (GOGAT) were investigated in Corynebacterium glutamicum 13032 . Under carbon-sufficient, nitrogen-limiting conditions, GS and GOGAT activities were five- and seven-fold higher, respectively, and transcription of the corresponding genes (glnA and gltBD) was similarly induced . GS activity was also induced in complete medium with added glucose, while GOGAT activity was unaffected . Under carbon-limiting, nitrogen-limiting conditions, the level of GS induction was reduced approximately three-fold, whereas GOGAT activity did not respond . Disruption of the hkm gene, encoding a putative histidine kinase upstream of gltBD, reduced the levels of GOGAT activity two-fold under both nitrogen-rich and nitrogen-limiting conditions . Promoter studies using a hkm-chloramphenicol acetylase fusion plasmid revealed that transcription of hkm is moderately induced (ca . 1.5-fold) by nitrogen starvation, indicating that the Hkm protein may play a role in signal transduction of the nutritional status of the growth medium. FEMS Microbiol Lett, 2001 Dec 18, 205(2), 277 - 82 Taxonomic characterization of two rubber degrading bacteria belonging to the species Gordonia polyisoprenivorans and analysis of hyper variable regions of 16S rDNA sequences; Arenskotter M et al.; Two cis-1,4-polyisoprene (isoprene rubber) degrading bacteria, strains VH2 and Y2K, were identified as strains of the species Gordonia polyisoprenivorans belonging to the Corynebacterineae, a suborder of the order Actinomycetales . Both showed characteristic growth and degradation of isoprene rubber as described previously for the type strain of G . polyisoprenivorans Kd2 (DSM 44302(T)) . For strain VH2 the chemotaxonomic properties were investigated, and DNA-DNA hybridization experiments with the type strain revealed the affiliation to the species G . polyisoprenivorans . The comparison of the 16S rDNA sequences, and especially hyper variable regions of these, led to the classification of strain Y2K to the same species . At present, the species G . polyisoprenivorans comprises three different isolates which share the ability to degrade isoprene rubber potently but which were obtained from different geographic regions. Biochim Biophys Acta, 2001 Dec 3, 1522(2), 138 - 41 Characterization of the cell surface protein gene of Corynebacterium ammoniagenes; Usuda Y et al.; Three dominant cell surface proteins of Corynebacterium ammoniagenes ATCC 6872 were identified in the cell wall fraction . The cspA gene, which encodes one of the major cell surface proteins, was cloned using the N-terminal amino acid sequence of the protein . Then the cloned chromosomal fragment containing the cspA gene was sequenced and was shown to encode a mature polypeptide of 333 amino acids with a molecular mass of 36654 Da . The amino acid sequence of the cspA gene showed similarity to the amino acid sequence of C . glutamicum CspA, one of the two major secreted proteins of C . glutamicum, although C . ammoniagenes CspA and C . glutamicum CspA differed in size . Northern blot analysis and primer extension analysis respectively revealed a 1.1 kb transcript and a promoter sequence resembling that of the C . ammoniagenes fatty acid synthase B (fasB) gene. Emerg Infect Dis, 2002 Jan, 8(1), 97 - 9 Nosocomial endocarditis caused by Corynebacterium amycolatum and other nondiphtheriae corynebacteria; Knox KL et al.; The nondiphtheriae corynebacteria are uncommon but increasingly recognized as agents of endocarditis in patients with underlying structural heart disease or prosthetic-valves . We describe three cases of nosocomial endocarditis caused by nondiphtheriae corynebacteria, including the first reported case of Corynebacterium amycolatum, endocarditis . These all occurred in association with indwelling intravascular devices. Ann Agric Environ Med, 2001, 8(2), 191 - 9 Exposure to airborne microorganisms in fiberboard and chipboard factories; Dutkiewicz J et al.; Microbiological air sampling was performed in one fiberboard factory and two chipboard factories located in south-eastern Poland . It was found that the levels of bacteria, fungi, dust and bacterial endotoxin in the air of examined facilities were high during initial stages of the production cycle (shredding of waste wood, storing of chips) and then sharply decreased during further stages of this cycle (forming and formatting of the boards) . In the fiberboard factory, the concentration of airborne microorganisms at the initial stages of production cycle was 71.8-95.2 x 10(3) cfu/m3 and dropped in further stages to the level of 8.4-17.5 x 10(3) cfu/m3 . Fungi (mostly Aspergillus fumigatus and Penicillium spp.) were prevailing microorganisms in the air of the fiberboard factory, forming 46.0-87.3% of the total airborne microflora . The concentrations of microorganisms in the air of the chipboard factories were significantly lower compared to the fiberboard factory (p<0.05) . During initial stages of production cycle they were within the range of 12.9-101.5 x 10(3) cfu/m3, while during forming and formatting of boards within the range of 5.3-12.4 x 10(3) cfu/m3 . On average, the most common microorganisms in the air of the chipboard factories were corynebacteria (mostly Arthrobacter spp . and Corynebacterium spp.) which formed 24.4-64.6% of the total microflora . The values of the respirable fraction of airborne microflora in the fiberboard and chipboard factories varied within a fairly wide range and were between 20.5-91.1% . Altogether, 38 species or genera of bacteria and 16 species or genera of fungi were identified in the air of examined factories, of which respectively 14 and 9 species or genera were reported as having allergenic and/or immunotoxic properties . The concentration of bacterial endotoxin in the air of examined factories was greatest, similarly to the concentration of microorganisms, during the initial stages of the production cycle: 103.1-1974.0 EU/m3 in the fiberboard factory, and 3.2-217.4 EU/m3 in chipboard factories . In conclusion, the workers of fiberboard and chipboard factories may be exposed during the initial stages of the production cycle (shredding of waste wood, storing of chips) to high levels of airborne microorganisms and endotoxin posing respiratory hazard. Biochem Biophys Res Commun, 2001 Dec 21, 289(5), 1307 - 13 The ptsI gene encoding enzyme I of the phosphotransferase system of Corynebacterium glutamicum; Kotrba P et al.; The phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) is widespread among bacteria where it mediates carbohydrate uptake and often serves in carbon control . Here we present cloning and analysis of the monocistronic ptsI gene of Corynebacterium glutamicum R, which encodes PTS Enzyme I (EI) . EI catalyzes the first reaction of PTS and the reported ptsI was shown to complement the corresponding defect in Escherichia coli . The deduced 59.2-kDa EI of 564 amino acids shares more than 50% homology with EIs from Bacillus stearothermophilus, Bacillus subtilis, and Lactobacillus sake . Chromosomal inactivation of ptsI demonstrated that EI plays an indispensable role in PTS of C . glutamicum R and this system represents a dominant sugar uptake system . Cellobiose was only transported and utilized in adaptive mutants of C . glutamicum R . Cellobiose transport was also found to be PTS-dependent and repressed by PTS sugar glucose. Endoscopy, 2001 Dec, 33(12), 1065 - 9 Abdominal actinomycosis: complication of endoscopic stenting in chronic pancreatitis? Harsch IA, Benninger J, Niedobitek G, Schindler G, Schneider HT, Hahn EG, Nusko G. Pancreatic endotherapy is frequently performed in patients with chronic pancreatitis and stenoses of the main pancreatic duct . In a patient with long-standing chronic pancreatitis and treatment with pancreatic stents, metastatic pancreatic head carcinoma was suspected because of infiltration of the neighboring organs and hepatic lesions . Ultrasound-guided aspiration of one liver lesion revealed grains typical for actinomycosis . In the light of this case, an extracted pancreatic stent was microbiologically investigated for actinomycetes in another patient who had a suspicious lesion of the pancreatic head . Microbiological examination of the extracted pancreatic stent revealed colonization by Actinomyces meyeri, Klebsiella oxytoca, and mixed cultures of anaerobic and saprophytic Gram-positive bacteria . In the following weeks, she developed a septic clinical picture with multiple abscesses of the liver . Actinomyces meyeri, Corynebacterium species, Candida and Enterococcae were cultivated in the aspirates . It seems possible, that treatment with pancreatic stents could have caused invasion of actinomycetes into the parenchyma of the pancreas, which was already harmed by the chronic inflammation, followed by the typical infiltrative growth and hematologic or biliary seeding into the liver. Int J Antimicrob Agents, 2001 Dec, 18(6), 571 - 4 Antimicrobial susceptibility of corynebacteria isolated from ewe's mastitis; Fernandez EP et al.; The antimicrobial susceptibility of 50 coryneform isolates from subclinical mastitis in sheep was evaluated . Arcanobacterium pyogenes (five isolates) had a susceptibility pattern distinct from the Corynebacterium species tested . The Corynebacterium isolates could be divided in two groups according to the MIC values for ciprofloxacin . Their antimicrobial susceptibility was usually unpredictable and consequently antimicrobial susceptibility tests are necessary for clinical and epidemiological purposes. Plasmid, 2001 Nov, 46(3), 153 - 62 Identification of a novel gene involved in stable maintenance of plasmid pGA1 from Corynebacterium glutamicum; Venkova T et al.; The cryptic plasmid pGA1 (4.8 kb) from Corynebacterium glutamicum, replicating in the rolling-circle mode, has been reported to contain four open reading frames longer than 200 bp (ORFA/per, ORFA2, ORFB, ORFC/rep) . Here we present another pGA1 gene, ORFE (174 bp), located in the region downstream of the per-ORFA2 gene cluster . The ORFE is transcribed into two RNA species in a direction opposite to that of the per-ORFA2 RNA . Introduction of ORFE in trans into the cells harboring the pGA1 derivatives carrying the main stability determinant, the per gene coding for a product that positively influences the pGA1 copy number and maintenance, increased their segregational stability . Mutation of the putative translational start of the ORFE abolished this observed positive effect in trans . ORFE thus codes for a protein acting as an accessory element involved in stable maintenance of plasmid pGA1 and was hence designated the aes gene (accessory effector of stable maintenance) . Vet Microbiol, 2002 Jan 23, 84(3), 253 - 61 Development of a Brucella suis specific hybridisation probe and PCR which distinguishes B . suis from Brucella abortus; Fayazi Z et al.; A genomic library was prepared from Brucella suis DNA (MboI digested) and cloned into the BamHI site of pUC18 . Colony hybridisation using a probe prepared from purified B . suis DNA labelled with alpha 32P was carried out to identify colonies of interest . About 20 colonies, which gave an intense signal upon hybridisation with whole B . suis genomic DNA as a probe, were selected . Because of the high degree of DNA homology between B . suis and Brucella abortus, a short probe was chosen as it would more likely give species specificity . Of seven fragments selected to probe whole B . suis, B . abortus, and Yersinia enterocolitica DNA, one was found to hybridise with B . suis only . The probe was sequenced in two directions and sense and anti sense primers of 25bp in length were chosen to yield a product of 421bp . After optimisation of the PCR, a product of 420bp was obtained with B . suis template DNA and two bands of 420 and 650bp were detected with B . abortus template DNA . This is the first reported PCR of the Brucella genome where a single pair of primers will discriminate between B . suis and B . abortus . No band was observed when the two primers were used to amplify E . coli, Y . enterocolitica, Enterobacter cloacae, Staphylococcus aureus, Streptococcus uberis, Corynebacterium bovis, or Serratia marcescens template DNA. FEMS Microbiol Lett, 2001 Nov 13, 204(2), 271 - 6 NADH dehydrogenase of Corynebacterium glutamicum . Purification of an NADH dehydrogenase II homolog able to oxidize NADPH; Matsushita K et al.; NADPH oxidase activity, in addition to NADH oxidase activity, has been shown to be present in the respiratory chain of Corynebacterium glutamicum . In this study, we tried to purify NADPH oxidase and NADH dehydrogenase activities from the membranes of C . glutamicum . Both the enzyme activities were simultaneously purified in the same fraction, and the purified enzyme was shown to be a single polypeptide of 55 kDa . The N-terminal sequence of the enzyme was consistent with the sequence deduced from the NADH dehydrogenase gene of C . glutamicum, which has been sequenced and shown to be a homolog of NADH dehydrogenase II . In addition to high NADH-ubiquinone-1 oxidoreductase activity at neutral pH, the purified enzyme showed relatively high NADPH oxidase and NADPH-ubiquinone-1 oxidoreductase activities at acidic pH . Thus, NADH dehydrogenase of C . glutamicum was shown to be rather unique in having a relatively high reactivity toward NADPH. Toxicology, 2001 Dec 28, 169(3), 195 - 208 Concurrent inflammation as a determinant of susceptibility to toxicity from xenobiotic agents; Ganey PE et al.; Sensitivity to the toxic effects of xenobiotic agents is influenced by a number of factors . Recent evidence derived from studies using experimental animals suggests that inflammation is one of these factors . For example, induction of inflammation by coexposure to bacterial endotoxin, vitamin A or Corynebacterium parvum increases injury in response to a number of xenobiotic agents that target liver . These agents are diverse in chemical nature and in mechanism of hepatotoxic action . Factors critical to the augmentation of liver injury by inflammation include Kupffer cells, neutrophils, cytokines such as tumor necrosis factor-alpha (TNF-alpha) and lipid mediators such as prostaglandins, but these may vary depending on the xenobiotic agent and the mechanisms by which it alters hepatocellular homeostasis . In addition, the timing of inflammagen exposure can qualitatively alter the toxic response to chemicals . Inflammation-induced increases in susceptibility to toxicity are not limited to liver . Concurrent inflammation also sensitizes animals to the toxic effects of agents that damage the respiratory tract, kidney and lymphoid tissue . It is concluded that inflammation should be considered as a determinant of susceptibility to intoxication by xenobiotic exposure. Arch Microbiol, 2001 Nov, 176(5), 381 - 5 Purification and characterization of an enzyme from Mycobacterium sp . Pyr-1, with nitroreductase activity and an N-terminal sequence similar to lipoamide dehydrogenase; Rafii F et al.; Mycobacterium sp . Pyr-1 produces an enzyme with nitroreductase activity that reduces 1-nitropyrene and 4-nitrobenzoic acid to the corresponding aromatic amines . This enzyme was constitutive and required NADH; and its activity was enhanced by FAD . It was inhibited by antimycin A, dicumarol, and o-iodosobenzoic acid; and it was inactivated by ammonium sulfate precipitation . After purification to homogeneity, the protein produced a single band on native and SDS-polyacrylamide gels and had a single amino-terminal sequence . The N-terminal amino acid sequence was identical to the corresponding sequences of the lipoamide dehydrogenases of M . leprae, M . tuberculosis and Corynebacterium glutamicum . The amino-terminal sequence was also similar to lipoamide dehydrogenases from M . smegmatis and several other bacteria . The amino acid sequence of an internal peptide (12 of 13 amino acids) was nearly identical to the corresponding sequences of lipoamide dehydrogenases from M . leprae and M . tuberculosis and was similar to those of C . glutamicum, Streptomyces coelicolor and S . seoulensis . The data show that a unique lipoamide dehydrogenase in Mycobacterium sp . Pyr-1, which differs from classic (Type I) bacterial nitroreductases, reduces aromatic nitro compounds to aromatic amines. Microbiology, 2001 Nov, 147(Pt 11), 2961 - 70 Glutamate synthase of Corynebacterium glutamicum is not essential for glutamate synthesis and is regulated by the nitrogen status; Beckers G et al.; The Corynebacterium glutamicum gltB and gltD genes, encoding the large (alpha) and small (beta) subunit of glutamate synthase (GOGAT), were investigated in this study . Using RT-PCR, a common transcript of gltB and gltD was shown . Reporter gene assays and Northern hybridization experiments revealed that transcription of this operon depends on nitrogen starvation . The expression of gltBD is under control of the global repressor protein AmtR as demonstrated by gel shift experiments and analysis of gltB transcription in an amtR deletion strain . In contrast to other bacteria, in C . glutamicum GOGAT plays no pivotal role; e.g . gltB and gltD inactivation did not result in growth defects when cells were grown in standard minimal medium and only a slight increase in the doubling time of the corresponding mutant strains was observed in the presence of limiting amounts of ammonia or urea . Additionally, mutant analyses revealed that GOGAT has no essential function in glutamate production by C . glutamicum. J Med Microbiol, 2001 Nov, 50(11), 1006 - 12 International external quality assessment scheme for the laboratory diagnosis of diphtheria; Engler KH et al.; An international external quality assessment (EQA) scheme has been established so as to evaluate the proficiency of specialist, national diphtheria reference laboratories in the laboratory diagnosis of diphtheria . Six simulated clinical specimens were freeze-dried and distributed to 23 participants in 20 countries . Participants were asked to isolate, identify and perform toxigenicity testing on any corynebacteria present and to complete a simple questionnaire describing the procedures and reagents used . Only three laboratories obtained correct biochemical and toxigenicity results for all six specimens . The majority of laboratories performed better with toxigenicity testing than with the biochemical identification . Of concern were the results from three laboratories that failed to isolate any corynebacteria from four or more of the specimens . In one centre this was shown to be due to lack of availability of 'in-date' media and, in the other two, was presumed to be due to lack of experience in primary laboratory diagnostics for this organism . It is essential that countries, globally, maintain awareness and laboratory capabilities in this specialised area of microbiology . EQA is an invaluable process, which enables laboratories to monitor, evaluate and improve their own performance in such areas. BMC Biotechnol . 2001;1(1):9 . Epub 2001 Oct 16. L-glutamate production by lysozyme-sensitive Corynebacterium glutamicum ltsA mutant strains; Hirasawa T et al.; BACKGROUND: A non-pathogenic species of coryneform bacteria, Corynebacterium glutamicum, was originally isolated as an L-glutamate producing bacterium and is now used for fermentative production of various amino acids . A mutation in the C . glutamicum ltsA gene caused susceptibility to lysozyme, temperature-sensitive growth, and L-glutamate production . RESULTS: The characteristics of eight lysozyme-sensitive mutants which had been isolated after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis were examined . Complementation analysis with the cloned wild-type ltsA gene and DNA sequencing of the ItsA region revealed that four mutants had a mutation in the ltsA gene . Among them, two mutants showed temperature-sensitive growth and overproduced L-glutamate at higher temperatures, as well as the previously reported ltsA mutant . Other two showed temperature-resistant growth: one missense mutant produced L-glutamate to some extent but the other nonsense mutant did not . These two mutants remained temperature-resistant in spite of introduction of ltsA::kan mutation that causes temperature-sensitive growth in the wild-type background . CONCLUSIONS: These results indicate that a defect caused by the ltsA mutations is responsible for temperature-sensitive growth and L-glutamate overproduction by C . glutamicum . The two temperature-resistant mutants seem to carry suppressor mutations that rendered cells temperature-resistance and abolished L-glutamate overproduction. MMW Fortschr Med, 2001 Oct 4, 143(40), 28 - 32 {Suspected diphtheria . Immediately start therapy!}; Stuck B et al.; Only those strains of corynebacteria that carry the gene for diphtheria toxin may cause diphtheria . The only known reservoir of C . diphtheriae is man . The past decade has seen a return of diphtheria in the newly independent states of the former Soviet Union . Owing to a lack of immunization of the population, more than 150,000 people went down with the disease . In Germany, too, contact resulted in a number of cases and two deaths . The sole effective protection is immunization . Although more than 90% of children and adolescents are protected, only 40% to 60% of adults are. FASEB J, 2001 Nov, 15(13), 2355 - 64 Protein tyrosine nitration in mouse peritoneal macrophages activated in vitro and in vivo: evidence against an essential role of peroxynitrite; Pfeiffer S et al.; Tyrosine nitration is considered a key reaction of peroxynitrite-triggered tissue injury in inflammatory diseases . We investigated the potential involvement of peroxynitrite in protein tyrosine nitration in isolated murine peritoneal macrophages activated either in vitro with interferon-gamma/lipopolysaccharide or in vivo by priming mice with Corynebacterium parvum (10 mgxkg-1) . Both protocols led to release of NO and accumulation of nitrite accompanied by formation of protein-bound 3-nitrotyrosine . Oxidation of dihydrorhodamine 123, a measure of peroxynitrite release, remained close to basal levels upon in vitro activation of the macrophages but was increased approximately twofold in vivo . Tyrosine nitration in macrophages activated in vitro was inhibited by catalase and the time course of nitration correlated with nitrite accumulation, whereas superoxide (O2*-) and H2O2 release occurred at much earlier times . To address the contribution of O2*- and peroxynitrite to in vivo nitration, a O2*- scavenger (MnTBAP; 1 mgxkg-1) was given to C . parvum-primed mice . MnTBAP led to almost complete inhibition of C . parvum-triggered O2*- and peroxynitrite release, whereas nitrite accumulation and formation of 3-nitrotyrosine were less affected ( approximately 50% of controls) . These results argue against an essential role of peroxynitrite in protein tyrosine nitration in vivo. Mol Immunol, 2001 Sep, 38(5), 397 - 408 Influence of relative binding affinity on efficacy in a panel of anti-CD3 scFv immunotoxins; Hexham JM et al.; The in vitro cell killing potency of an immunotoxin reflects the aggregate of several independent biochemical properties . These include antigen binding affinity; internalization rate, intracellular processing and intrinsic toxin domain potency . This study examines the influence of antigen binding affinity on potency in various immunotoxin fusion proteins where target antigen binding is mediated by single chain antibody variable region fragments (scFv) . Firstly, the relationship between affinity and potency was examined in a panel of four scFv immunotoxins generated from different anti-CD3 monoclonal antibodies fused to the 38 kDa fragment of Pseudomonas aeruginosa exotoxin A (PE38) . Of these four scFv-PE38 immunotoxins, the one derived from the anti-CD3 monoclonal antibody UCHT1 has highest cell killing potency . Analysis of these four scFv-PE38 immunotoxins indicated a correlation between antigen binding affinity and immunotoxin potency in the cell killing assay with the exception of the scFvPE38 immunotoxin derived from the antibody BC3 . However this scFv appeared to suffer a greater drop in affinity ( approximately 100x), relative to the parent Mab than did the other three scFvs used in this study (2-10x) . Secondly, the scFv(UCHT1)-PE38 immunotoxin was then compared with a further panel of scFv(UCHT1)-derived immunotoxins including a divalent PE38 version and both monovalent and divalent Corynebacterium diphtheriae toxin (DT389) fusion proteins . When the scFv-UCHT1 domain was amino-terminally positioned relative to the toxin, as in the scFv(UCHT1)-PE38, an approximately 10-fold higher antigen-binding affinity was observed than with the C-terminal fusion, used in the DT389-scFv(UCHT1) molecule . Despite this lower antigen-binding activity, the DT389-scFv immunotoxin had a 60-fold higher potency in the T-cell-killing assay . Thirdly, a divalent form of the DT389-scFv construct, containing tandem scFv domains, had a 10-fold higher binding activity, which was exactly reflected in a 10-fold increase in potency . Therefore, when comparing immunotoxins in which scFvs from different antibodies are fused to the same toxin domain (DT or PE) a broad correlation appears to exist between binding affinity and immunotoxin potency . However, no correlation between affinity and potency appears to exist when different toxin domains are combined with the same scFv antibody domain. Metab Eng, 2001 Oct, 3(4), 344 - 61 Metabolic consequences of altered phosphoenolpyruvate carboxykinase activity in Corynebacterium glutamicum reveal anaplerotic regulation mechanisms in vivo; Petersen S et al.; Corynebacterium glutamicum possesses high in vivo activity of the gluconeogenic phosphoenolpyruvate carboxykinase (PEPCk) during growth on glucose, resulting together with anaplerotic carboxylation reactions in a PEP/pyruvate/oxaloacetate substrate cycle . The present study investigated the changes in intracellular fluxes and metabolite concentrations that are caused by altered PEPCk activity in L-lysine-producing C . glutamicum MH20-22B, applying a recently developed (13)C labeling-based strategy for anaplerotic flux resolution and quantification . Abolition of PEPCk activity by deletion of the respective pck gene resulted in increased intracellular concentrations of oxaloacetate L-aspartate, alpha-ketoglutarate, pyruvate, and L-lysine and in a 60% enhanced flux toward L-lysine biosynthesis, whereas increasing the PEPCk activity by pck overexpression had opposite effects . The results of the combined measurements of enzyme activities, in vivo fluxes, and metabolite concentrations were exploited to elucidate the in vivo regulation of anaplerotic reactions in C . glutamicum, and implications for the metabolic engineering of amino-acid-producing strains are discussed . Eur J Pharmacol, 2001 Aug 3, 425(1), 73 - 83 Modulation of cytokine responses in Corynebacterium parvum-primed endotoxemic mice by centrally administered cannabinoid ligands; Smith SR et al.; The cannabinoid receptor agonists {(-)-11-hydoxy-Delta(8)tetrahydrocannabinol-dimethylheptyl} (HU-210) and {(R)-(+)-{2,3-dihydro-5-methyl-3-{(4-morpholinyl)methyl{pyrrolo{1,2,3-de}1,4-benzoxazin-6-yl}(1-naphthalenyl) methanone} (WIN 55212-2) were previously shown to downregulate inflammatory cytokines (tumor necrosis factor alpha and interleukin-12) and to upregulate antiinflammatory interleukin-10 when administered intraperitoneally (i.p.) to mice before an endotoxin challenge . Cytokine modulation coincided with the onset of behavioral changes that are associated with cannabinoid agonist activated central cannabinoid CB(1) receptors . Both effects were antagonized by {N-(piperdin-1-yl)-5-(4-chloropheny)-1-(2,4-dichloropheny)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride} (SR141716A) a selective cannabinoid CB(1) receptor antagonist . In the present study, we have investigated further the apparent role of central CB(1) cannabinoid receptors in cytokine modulation by HU-210 and WIN 55212-2 . When administered intracerebroventricularly (i.c.v.), the drugs modulated cytokine responses at doses that were threefold to fourfold lower than those found effective by the i.p . route . SR141716A blocked cytokine modulation when coadministered centrally with the agonists, while a selective cannabinoid CB(2) receptor antagonist, (N-{(1S)-endo-1,3,3-trimethylbicyclo{2.2.1}heptan-2-yl}5-(4-choro-3 methylphenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide) (SR144528) had no effect . Surprisingly, SR144528 was found to modulate cytokines itself when injected i.c.v. Aviakosm Ekolog Med, 2001, 35(4), 32 - 40 {Basic patterns of microflora development in the environment of orbital complex Mir}; Novikova ND; During the multi-year operation of orbital complex Mir a systematic survey of development and behavior of microflora was performed in order to assess the associated risks to the piloted vehicle . Microorganisms isolated from the Mir environment represented 234 species of which 108 were bacteria and 126 microscopic fungi . Bacteria included opportunistic pathogens causing opportunistic infections . There were also fungi that were pathogenic for humans leading to mycosis and mycotoxication . The largest species diversity was characteristic of fungi, so-called technophils damaging polymers and corroding metals . Staphylococcus, Micrococcus, Corynebacterium Bacillus dominated among the bacteria and Penicullium, Aspergillus and Cladosporium were most common, omnipresent and numerous among the fungi . Microbial contamination of Mir undulated so that activation periods alternated period of stabilization and other species prevailed in number and spread . Evaluation of microflora in this environment was conducive of medical and technological risks. Curr Opin Biotechnol, 2001 Oct, 12(5), 446 - 9 Expression systems for use in actinomycetes and related organisms; Connell ND; There have been significant advances in genetic and molecular approaches to understanding the physiology of organisms belonging to the genera Mycobacterium, Corynebacterium, Nocardia and Streptomyces . This review discusses recent advances in heterologous protein expression in members of the actinomycete group, including codon usage, post-translational modification and inducible gene expression. J Food Prot, 2001 Oct, 64(10), 1584 - 91 Effects of storage temperature and preservative treatment on shelf life of the pond-raised freshwater fish, silver perch (Bidyanus bidyanus); Gelman A et al.; Sensory and microbiological characteristics of pond-raised freshwater silver perch (Bidyanus bidyanus) fish, during cold storage over a period of 25 days were evaluated . Whole fish (averaging 400 g each) were stored in cold storage rooms at either 0 to 2 degrees C, 5 degrees C, or 5 degrees C + potassium sorbate as a preservative . The organoleptic and hypoxanthine test results show that the treatment of potassium sorbate can slow the process of spoilage by about 5 days . Yet, the most important factor affecting the shelf life of these fish is the storage temperature . Keeping the fish at 0 to 2 degrees C can prolong the storage prior to spoilage by 10 days compared with those kept at 5 degrees C . These results obtained through organoleptic tests are corroborated by both the chemical (hypoxanthine and total volatile basic nitrogen) and to some extent by the physical (cosmos) tests . The initial total bacteriological counts were 5 x 10(2) CFU/cm2 for fish surface and <10(2) CFU/g for fish flesh, and these counts rose continuously, reaching about 106 CFU/g (0 to 2 degrees C) and 10(7) CFU/g (5 degrees C) in flesh and 10(7) to 10(8) CFU/cm2 on the surface by the end of the storage period . The addition of potassium sorbate led to a smaller increase in bacterial numbers, especially during the first 15 days . Bacterial composition fluctuated during storage . The initial load on the fish surface was predominantly mesophilic and gram positive and consisted mostly (80%) of Micrococci, Bacillus, and Corynebacterium . During the next 10 days, these bacteria were practically replaced by gram-negative flora comprised mostly of Pseudomonas fluorescens that rapidly increased with storage time and accounted for 95% after 15 days. Appl Microbiol Biotechnol, 2001 Sep, 56(5-6), 710 - 7 Effect of transketolase modifications on carbon flow to the purine-nucleotide pathway in Corynebacterium ammoniagenes; Kamada N et al.; Transketolase, one of the enzymes in the nonoxidative branch of the pentose phosphate pathway, operates to shuttle ribose 5-phosphate and glycolytic intermediates together with transaldolase, and might be involved in the availability of ribose 5-phosphate, a precursor of nucleotide biosynthesis . The tkt and tal genes encoding transketolase and transaldolase, respectively, were cloned from the typical nucleotide- and nucleoside-producing organism Corynebacterium ammoniagenes by a PCR approach using oligonucleotide primers derived from conserved regions of each amino acid sequence from other organisms . Enzymatic and molecular analyses revealed that the two genes were clustered on the genome together with the glucose 6-phosphate dehydrogenase gene (zwf) . The effect of transketolase modifications on the production of inosine and 5'-xanthylic acid was investigated in industrial strains of C . ammoniagenes . Multiple copies of plasmid-borne tkt caused about tenfold increases in transketolase activity and resulted in 10-20% decreased yields of products relative to the parents . In contrast, site-specific disruption of tkt enabled both producers to accumulate 10-30% more products concurrently with a complete loss of transketolase activity and the expected phenotype of shikimate auxotrophy . These results indicate that transketolase normally shunts ribose 5-phosphate back into glycolysis in these biosynthetic processes and interception of this shunt allows cells to redirect carbon flux through the oxidative pentose pathway from the intermediate towards the purine-nucleotide pathway. Genome Biol . 2001;2(10):RESEARCH0044 . Epub 2001 Sep 19. The ESAT-6 gene cluster of Mycobacterium tuberculosis and other high G+C Gram-positive bacteria; Gey Van Pittius NC et al.; BACKGROUND: The genome of Mycobacterium tuberculosis H37Rv has five copies of a cluster of genes known as the ESAT-6 loci . These clusters contain members of the CFP-10 (lhp) and ESAT-6 (esat-6) gene families (encoding secreted T-cell antigens that lack detectable secretion signals) as well as genes encoding secreted, cell-wall-associated subtilisin-like serine proteases, putative ABC transporters, ATP-binding proteins and other membrane-associated proteins . These membrane-associated and energy-providing proteins may function to secrete members of the ESAT-6 and CFP-10 protein families, and the proteases may be involved in processing the secreted peptide . RESULTS: Finished and unfinished genome sequencing data of 98 publicly available microbial genomes has been analyzed for the presence of orthologs of the ESAT-6 loci . The multiple duplicates of the ESAT-6 gene cluster found in the genome of M . tuberculosis H37Rv are also conserved in the genomes of other mycobacteria, for example M . tuberculosis CDC1551, M . tuberculosis 210, M . bovis, M . leprae, M . avium, and the avirulent strain M . smegmatis . Phylogenetic analyses of the resulting sequences have established the duplication order of the gene clusters and demonstrated that the gene cluster known as region 4 (Rv3444c-3450c) is ancestral . Region 4 is also the only region for which an ortholog could be found in the genomes of Corynebacterium diphtheriae and Streptomyces coelicolor . CONCLUSIONS: Comparative genomic analysis revealed that the presence of the ESAT-6 gene cluster is a feature of some high-G+C Gram-positive bacteria . Multiple duplications of this cluster have occurred and are maintained only within the genomes of members of the genus Mycobacterium. Clin Nephrol, 2001 Sep, 56(3), 241 - 6 Tsukamurella peritonitis associated with continuous ambulatory peritoneal dialysis; Shaer AJ et al.; A case of Tsukamurella peritonitis associated with peritoneal dialysis in a 23-year-old woman is described . The organism was difficult to identify and was mistaken for Corynebacterium and atypical mycobacteria . Despite prolonged, multidrug, antimicrobial therapy with conventional antibiotics including vancomycin, ciprofloxacin, rifampin, gentamicin and ceftazidime, catheter removal was required to successfully treat peritonitis . Human infection due to this organism is rare and has been previously reported in only 13 cases, 1 of which was peritonitis . We describe here the second case of Tsukamurella peritonitis associated with peritoneal dialysis. Int J Syst Evol Microbiol, 2001 Sep, 51(Pt 5), 1723 - 8 Corynebacterium freneyi sp . nov., alpha-glucosidase-positive strains related to Corynebacterium xerosis; Renaud FN et al.; Three coryneform strains from clinical specimens were studied . They belonged to the genus Corynebacterium, since they had type IV cell walls containing corynemycolic acids . They had phenotypic characteristics that included alpha-glucosidase, pyrazinamidase and alkaline phosphatase activities and fermentation of glucose, ribose, maltose and sucrose . These are the characteristics of Corynebacterium xerosis . Since this species is very rare in human pathology, the strains were studied in more detail by comparing the 16S-23S intergenic spacers, rDNA sequences and levels of DNA similarity of these three strains and those of the reference strains C . xerosis ATCC 373T and Corynebacterium amycolatum CIP 103452T . According to DNA-DNA hybridization data, the three novel strains are members of the same species (level of DNA similarity >72%) . Phylogenetic analysis revealed that these strains are closely related to C . xerosis and C . amycolatum, but DNA-relatedness experiments showed clearly that they constitute a distinct new species, with levels of DNA relatedness of less than 23% to C . xerosis ATCC 373T and less than 5% to C . amycolatum CIP 103452T . Two other alpha-glucosidase-positive strains presenting the same biochemical characteristics were included in the study and proved to be C . amycolatum . This new species can be differentiated from C . xerosis and C . amycolatum strains by carbon source utilization, intergenic spacer region length profiles and some biochemical characteristics such as glucose fermentation at 42 degrees C and growth at 20 degrees C . The name Corynebacterium freneyi sp . nov . is proposed with the type strain ISPB 6695110T (= CIP 106767T = DSM 44506T). Biodegradation, 2000, 11(6), 371 - 6 Growth rate influences reductive biodegradation of the organophosphorus pesticide demeton by Corynebacterium glutamicum; Girbal L et al.; The organophosphorous pesticide, demeton-S-methyl was transformed by Corynebacterium glutamicum in co-metabolism with more readily degradable substrates . Glucose, acetate and fructose were tested as growth substrates, and the highest demeton-S-methyl biotransformation average rate (0.78 mg l(-1) h(-1)) and maximum instantaneous rate (1.4 mg l(-1) h(-1)) were achieved on fructose . This higher efficiency seems to be linked to the atypical behavior of C . glutamicum grown on fructose, characterized by a prolonged period of accelerating growth instead of a constant growth rate observed on glucose or acetate . More precisely, for growth rates in the 0.1-0.4 h(-1) range, a direct coupling between the specific demeton-S-methyl consumption rate and the growth rate was demonstrated on fructose during batch-, steady state continuous- or continuous cultures with a controlled transient growth rate (accelerostat technology) . The demeton-S-methyl biotransformation was more favoured during an acceleration phase of the growth rate. Microbiology, 2001 Oct, 147(Pt 10), 2865 - 71 Cytochrome c oxidase contains an extra charged amino acid cluster in a new type of respiratory chain in the amino-acid-producing Gram-positive bacterium Corynebacterium glutamicum; Sakamoto J et al.; The membranes from Corynebacterium glutamicum cells contain a hydrophobic di-haem C protein as the cytochrome c subunit of the new type of cytochrome bc complex (complex III in the respiratory chain) encoded by the qcrCAB operon {Sone, N., Nagata, K., Kojima, H., Tajima, J., Kodera, Y., Kanamaru, T., Noguchi, S . & Sakamoto, J . (2001) . Biochim Biophys Acta 1503, 279-290} . To characterize complex IV, cytochrome c oxidase and its structural genes were isolated . The oxidase is of the cytochrome aa(3) type, but mass spectrometry indicated that the haem is haem As, which contains a geranylgeranyl side-chain instead of a farnesyl group . The enzyme is a SoxM-type haem-copper oxidase composed of three subunits . Edman degradation and mass spectrometry suggested that the N-terminal signal sequence of subunit II is cleaved and that the new N-terminal cysteine residue is diacylglycerated, while neither subunit I nor subunit III is significantly modified . The genes for subunits II (ctaC) and III (ctaE) are located upstream of the qcrCAB operon, while that for subunit I (ctaD) is located separately . The oxidase showed low enzyme activity with extrinsic substrates such as cytochromes c from horse heart or yeast, and has the Cu(A)-binding motif in its subunit II . A prominent structural feature is the insertion of an extra charged amino acid cluster between the beta2 and beta4 strands in the substrate-binding domain of subunit II . The beta2-beta4 loop of this oxidase is about 30 residues longer than that of major cytochrome c oxidases from mitochondria and proteobacteria, and is rich in both acidic and basic residues . These findings suggest that the extra charged cluster may play a role in the interaction of the oxidase with the cytochrome c subunit of the new type of bc complex. J Infect Dis, 2001 Oct 15, 184(8), 1035 - 40 Epub 2001 Aug 31. Diphtheria in Thailand in the 1990s; Tharmaphornpilas P et al.; Diphtheria remains endemic in developing countries, but there are limited published data on the subject . Thailand's diphtheria surveillance data are relatively complete and may give a fuller picture of the situation in similar countries . After routine immunization began in 1977, the incidence of reported diphtheria decreased by >98% to <0.1 case per 100,000 persons annually in the 1990s . Despite infant immunization coverage of >90%, diphtheria cases were reported throughout the 1990s, primarily among children <15 years old . Outbreaks were linked to both persistent endemic circulation and to importation of toxigenic Corynebacterium diphtheriae; suboptimal immunization coverage in minority and disadvantaged groups contributed . A serologic survey found 25% of adults 20-39 years old and 14% of adolescents 10-19 years old lacked immunity to diphtheria; these data indicate an accumulation of susceptible adolescents and adults . Diphtheria remains a threat in Thailand; improvements in diphtheria control will depend on improving childhood immunization coverage in Thailand and the surrounding region. EMBO J, 2001 Oct 1, 20(19), 5412 - 20 The osmoreactive betaine carrier BetP from Corynebacterium glutamicum is a sensor for cytoplasmic K+; Rubenhagen R et al.; The isolated glycine betaine uptake carrier BetP from Corynebacterium glutamicum was reconstituted in Escherichia coli phospholipid liposomes and its response to osmotic stress studied . The transport activity of BetP, which was previously shown to comprise both osmosensory and osmoregulatory functions, was used to identify the nature of the physicochemical stimulus related to hyperosmotic stress . Putative factors modulating transport activity in response to osmotic stress were dissected . These include type, osmolality and concentration of solutes in the internal and/or external compartment (cationic, anionic, zwitterionic, neutral), as well as membrane strain as a response to increased osmolality . Osmoresponsive activation of BetP was independent of any external factor and of physical alterations of the membrane, but was triggered by a change in the internal K+ concentration . Activation did not depend on the type of anion present and was K+ (or Cs+ and Rb+) specific, as choline and NH(4)+ did not trigger BetP activity . The half-maximal activation of BetP in E.coli phospholipid liposomes was correlated to an internal concentration of 221 +/- 23 mM K+. Chemosphere, 2001 Oct, 45(1), 45 - 50 Aerobic dehalogenation potentials of four bacterial species isolated from soil and sewage sludge; Olaniran AO et al.; Four bacterial species each were isolated from soil and a sewage oxidation pond using enrichment culture technique, and the bacterial isolates were identified to belong to the two genera Bacillus and Corynebacterium . The axenic cultures of the isolates utilized monochloroacetic acid (MCA), trichloroacetic acid (TCA), CHCl3 and CCl4 for growth up to 1 g substrate l(-1) (w/v) and growths were enhanced in the mixed cultures of the isolates . The specific growth rate constants in the media ranged from 0.144 to 0.475 . This is lower than the comparatively high values observed for the glucose medium, which varied significantly (P < 0.05) between 0.699 and 0.792 h(-1) . Serial adaptation of the individual isolates in the organochloride media significantly (P < 0.05) affected the bacteria growth yields . The dehalogenase specific activity observed in the cell-free extracts of the mixed cultures of the isolates was significantly higher (P < 0.05) than those of their respective monocultures . Optimal pH of the dehalogenase activity was between 7.6 and 8.0, while temperature optima were between 30 degrees C and 35 degrees C. Clin Infect Dis, 2001 Nov 1, 33(9), 1598 - 600 Epub 2001 Sep 24. Infection of the skin caused by Corynebacterium ulcerans and mimicking classical cutaneous diphtheria; Wagner J et al.; Extrapharyngeal infections caused by Corynebacterium ulcerans have rarely been reported previously, and diphtheria toxin production has usually not been addressed . This case demonstrates that strains of C . ulcerans that produce diphtheria toxin can cause infections of the skin that completely mimic typical cutaneous diphtheria, thereby potentially providing a source of bacteria capable of causing life-threatening diseases in the patient's environment . Therefore, it is recommended to screen wound swabs for coryneform bacteria, identify all isolates, carefully assess possible toxin production, and send questionable strains to a specialist or a reference laboratory. J Mol Microbiol Biotechnol, 2001 Oct, 3(4), 573 - 83 Characterization of the phosphoenolpyruvate carboxykinase gene from Corynebacterium glutamicum and significance of the enzyme for growth and amino acid production; Riedel C et al.; Corynebacterium glutamicum possesses phosphoenolpyruvate (PEP) carboxykinase, oxaloacetate decarboxylase and malic enzyme, all three in principle being able to catalyze the first step in gluconeogenesis . To investigate the role of PEP carboxykinase for growth and amino acid production, the respective pck gene was isolated, characterized and used for construction and analysis of mutants and overexpressing strains . Sequence analysis of the pck gene predicts a polypeptide of 610 amino acids showing up to 64% identity with ITP-/GTP-dependent PEP carboxykinases from other organisms . C . glutamicum cells harbouring pck on plasmid showed about tenfold higher specific PEP carboxykinase activities than the wildtype . Inactivation of the chromosomal pck gene led to the absence of PEP carboxykinase activity and the inability to grow on acetate or lactate indicating that the enzyme is essential for growth on these carbon sources and thus, for gluconeogenesis . The growth on glucose was not affected . Examination of glutamate production by the recombinant C . glutamicum strains revealed that the PEP carboxykinase-deficient mutant showed about fourfold higher, the pck-overexpressing strain two- to threefold lower glutamate production than the parental strain . Inactivation and overexpression of pck in a lysine-producer of C . glutamicum led to an only 20% higher and lower lysine accumulation, respectively . The results show that PEP carboxykinase activity in C . glutamicum is counteractive to the production of glutamate and lysine and indicate that the enzyme is an important target in the development of strains producing amino acids derived from citric acid cycle intermediates. Mol Cells, 2001 Aug 31, 12(1), 112 - 6 Gene lmrB of Corynebacterium glutamicum confers efflux-mediated resistance to lincomycin; Kim HJ et al.; The lmrB gene of Corynebacterium glutamicum, which confers specific resistance to lincosamides, such as lincomycin and clindamycin, was isolated . C . glutamicum cells, carrying the lmrB gene in a multicopy plasmid, showed increased resistance to lincomycin with a MIC of 230 microg/ml, which is a 9-fold increase compared to that of the wild type . The lmrB-disrupted mutant became sensitive to the compound . No difference in sensitivity to erythromycin, penicillin G, tetracycline, chloramphenicol, spectinomycin, nalidixic acid, gentamicin, streptomycin, ethidium bromide, and sodium dodecyl sulfate was observed . The protonophore carbonyl cyanide m-chlorophenylhydrazone abolished the lincomycin-resistance of lmrB-carrying cells . The putative protein product of the gene contained 14-transmembrane regions and showed high amino acid-sequence homology to the drug efflux pumps of other organisms . In addition, the putative protein contained a motif for major facilitators, suggesting a role in efflux-mediated resistance to lincomycin. Vet Rec, 2001 Aug 25, 149(8), 232 - 5 Conjunctival flora observed in 70 healthy domestic rabbits (Oryctolagus cuniculus); Cooper SC et al.; Conjunctival swabs were taken from both eyes of 70 healthy domestic rabbits and cultured to determine the microbial population . Bacteria were recovered from 83 per cent of the specimens . DNase-negative Staphylococcus species (57 per cent) were the most commonly recovered organisms followed by Micrococcus species (25 per cent) and Bacillus species (19 per cent) . Other organisms isolated included Stomatococcus species (8 per cent), Neisseria species (8 per cent), Pasteurella species (6 per cent), Corynebacterium species (6 per cent), Streptococcus species (6 per cent) and Moraxella species (4 per cent), and other bacteria were isolated less frequently . Statistical analysis showed that there appeared to be no significant difference between the bacterial isolation rates from different breeds of rabbit . Significantly more of the swabs taken from young rabbits yielded cultivable bacteria than did those taken from rabbits over 12 months of age. Eur J Anaesthesiol, 2001 Sep, 18(9), 599 - 604 Inflammatory liver disease shortens atracurium-induced neuromuscular blockade in rats; Mayer B et al.; BACKGROUND: and objective Inflammatory liver dysfunction in rats leads to a prolonged vecuronium-induced neuromuscular blockade due to insufficient metabolism . A coexisting resistance against the drug partly counteracts this prolongation . The present study investigates the pharmacodynamics of atracurium whose metabolism does not depend on liver function . METHODS: Male Sprague-Dawley rats (n=14; 290 +/- 30 g) were randomly allocated to either a group in which liver inflammation was induced by intravenous injection of 60 mg kg(-1) heat-killed Corynebacterium parvum or to a control group . On day 5 after injection, liver function was assessed using the aminopyrine breath test . Under propofol anaesthesia, duration of action of atracurium (4.8 mg kg(-1)) was measured by evoked mechanomyography (stimulation of the sciatic nerve; contraction of the gastrocnemius muscle) . Nitric oxide concentrations, as variables for the severity of the inflammation, were assessed by measurement of nitrite/nitrate plasma concentrations . RESULTS: In C . parvum-injected rats, nitrite/nitrate plasma concentrations were increased (972 +/- 597 vs . 25 +/- 7 micromol L(-1)), the aminopyrine turnover was depressed (1.7 +/- 0.4% vs . 3.5 +/- 0.5%), and the atracurium-induced neuromuscular blockade was shortened (372 +/- 128 s vs . 1081 +/- 234 s) . CONCLUSIONS: A systemic inflammatory response syndrome with liver dysfunction results in decreased sensitivity to atracurium . Further investigations are needed regarding a possible up-regulation of acetylcholine receptors or an increased protein binding of atracurium during sepsis to clarify reasons behind this phenomenon. Zh Mikrobiol Epidemiol Immunobiol, 2001 May-Jun, (3), 3 - 8 {Genetic structure of Corynebacterium diphtheriae strains isolated in Russia during epidemics of various intensity}; Kombarova SIu et al.; The genetic structure of C . dipthteriae toxigenic strains isolated in Russia during the period of more than 50 years was analysed . The use of the method of ribotyping made it possible to register 17 C . diphtheriae ribotypes . The study revealed that the genetic structure of C . diphtheriae population varied in the dynamics of the epidemic process: each epidemic cycle characterized by predominant spread of epidemic strains of definite biovars and ribotypes . Thus, C . diphtheriae strains of biovar gravis, ribotype M11, dominated in the 40-60 years and C . diphtheriae strains of biovar mitis, closely related ribotypes M1 and M1v, dominated in the 80 years . During the last epidemic rise of diphtheriae morbidity in the 90 s C . diphtheriae strains of biovar gravis, closely related ribotypes G1 and G4, dominated among circulating strains . The proportion of these ribotypes began to increase 3 years before the rise of morbidity . The data of microbiological monitoring are recommended for use in the prognostication of the development of the epidemic process of diphtheria infection. J Med Microbiol, 2001 Sep, 50(9), 795 - 804 Identification of two Mycobacterium avium subspecies paratuberculosis gene products differentially recognised by sera from rabbits immunised with live mycobacteria but not heat-killed mycobacteria; Bannantine JP et al.; The investigation of environmentally regulated proteins has led to a better understanding of host-pathogen interactions and identified novel vaccine candidate antigens for several bacterial pathogens . In an effort to identify such proteins in Mycobacterium avium subsp . paratuberculosis (M . paratuberculosis), a genomic expression library was differentially screened with sera from rabbits that had been immunised with live M . paratuberculosis (alpha-live) as well as sera from rabbits immunised with heat-killed M . paratuberculosis (alpha-killed) . These experiments identified seven recombinant plaques that were uniquely recognised by the alpha-live sera . Sequence data showed that five of these clones overlapped with each other and contained a common open-reading frame encoding a 25-kDa protein, termed Csp1 . The 25-kDa antigen shows weak similarity to a secreted Corynebacterium glutamicum protein . The remaining two clones overlapped with each other and contained two partial open-reading frames, both encoding proteins with strong homology to polyketide synthase from various species of mycobacteria . Antisera were produced against a peptide of the polyketide synthase gene product designated Pks7 . Csp1-specific antibodies were affinity purified from the alpha-live sera . These purified antibodies demonstrated that Csp1 was present within infected macrophages . Collectively, these data identify novel M . paratuberculosis antigens that may be important in pathogenesis. Zh Mikrobiol Epidemiol Immunobiol, 2001 Mar-Apr, (2), 83 - 5 {Identification of Corynebacterium diphtheria biovar belfanti among circulating C . diphtheriae strains in Ukraine}; Demikhovskaia EV et al.; The biochemical test of the reduction of nitrates to nitrites made it possible to identify 5.2% of strains belonging to biovar belfanti among 135 C . diphtheriae strains, initially classified within biovar mitis . Out of 7 identified C . diphtheriae belfanti strains, 2 toxigenic strains were isolated from multiple foci diphtheria . According to the results of the polymerase chain reaction, 1 out of 5 non-toxigenic strains had tox gene . All C . diphtheriae belfanti strains were found to have pronounced capacity for adhesion to sheep and human red blood cells . At the stage of the extinction of diphtheria epidemic the practical identification of C . diphtheriae belfanti strains is necessary, as increased adhesion in combination with toxigenic properties may probably promote for bacteria of this biovar to take the leading role at the period of sporadic morbidity. J Mol Microbiol Biotechnol, 2001 Oct, 3(4), 529 - 43 Regulation of aromatic amino acid biosynthesis in gamma-proteobacteria; Panina EM et al.; Computational comparative techniques were applied to analysis of the aromatic amino acid regulons in gamma-proteobacteria . This resulted in characterization of the TrpR and TyrR regulons in the genomes of Yersinia pestis, Haemophilus influenzae, Vibrio cholerae and other bacteria and identification of new members of the PhhR regulon in the genome of Pseudomonas aeruginosa . Candidate attenuators were constructed for all studied genomes, including the trpBA operon of the very distantly related bacterium Chlamidia trachomatis . The pheA attenuator of Y . pestis is an integration site for the insertion element IS-200 . It was shown that the triplication of the DAHP-synthase genes occurred prior to the divergence of families Enterobacteriaceae, Vibrionaceae and Alteromonadaceae . The candidate allosteric control site of the DAHP-syntheases was identified . This site is deteriorated in AroH of Buchnera sp . APS . The known DAHP-synthase of Bordetella pertussis is likely to be feedback-inhibited by phenylalanine, and the DAHP-synthase of Corynebacterium glutamicum could be inhibited by tyrosine . Overall, the most extensive regulation was observed in Escherichia coli, whereas the regulation in other genomes seems to be less developed . At the extreme, the tryptophan production in the aphid endosymbiont Buchnera sp . APS is free from transcriptional, attenuation, and allosteric control. Adv Space Res, 1992, 12(4), 255 - 63 Long-term preservation of microbial ecosystems in permafrost; Gilichinsky DA et al.; It has been established that significant numbers (up to 10 million cells per gram of sample) of living microorganisms of various ecological and morphological groups have been preserved under permafrost conditions, at temperatures ranging from -9 to -13 degrees C and depths of up to 100 m, for thousands and sometimes millions of years . Preserved since the formation of permafrost in sand-clay sediments of the Pliocene-Quaternary period and in paleosols and peats buried among them, these cells art the only living organisms that have survived for a geologically significant period of time . The complexity of the microbial community preserved varies with the age of the permafrost . Eukaryotes are found only in Holocene sediments; while prokaryotes are found to greater ages, i.e., Pliocene and Pleistocene . The diversity of microorganisms decreases with increasing age of sediments, and as a result cocci and corynebacteria are predominant . Enzyme activity (catalase and hydrolytic enzymes) and photosynthetic pigments (chlorophyll and pheophytin have also been detected in permafrost sediments . These results permit us to outline some approaches to the search for traces of life in the permafrost of Martian sediments by borehole core sampling . It is in the deep horizons (and not on the planet surface), isolated by permafrost from the external conditions, that results similar to those obtained on Earth can be expected. Adv Space Res, 1986, 6(12), 287 - 98 Microbial life at extremely low nutrient levels; Hirsch P; Many microorganisms ("oligotrophs") grow in distilled water: Pseudomonas spp., Caulobacter spp., Hyphomicrobium spp., Arthrobacter spp., Seliberia spp., Bactoderma alba, Corynebacterium spp., Amycolata (Nocardia) autotrophica, Mycobacterium spp., yeasts, and Chlorella spp . Also, certain lower fungi can be found here . In the laboratory, these organisms thrive on contaminations of the air (CO, hydrocarbons, H2, alcohols etc.) . All are euryosmotic and often grow also in higher concentrations of salts and nutrients . Natural locations with extremely low nutrient levels (snow, rain water pools, springs, free ocean water, Antarctic rocks and soils) do not contain more than 1-5 mg/l of organic carbon . Oligotrophs found here are especially adapted to constant famine: they frequently live attached to surfaces, form polymers and storage products even while starving, and often aggregate . Many of these oligotrophs alter their morphology (surface to volume ratio) with changing nutrient concentrations . Extreme oligotrophs also occur in generally nutrient-rich environments such as sewage aeration tanks or compost soil . Here they are thought to survive in nutrient-depauperate microhabitats. Appl Environ Microbiol, 2001 Sep, 67(9), 4293 - 304 Isolation of toxigenic Nocardiopsis strains from indoor environments and description of two new Nocardiopsis Species, N . exhalans sp . nov . and N . umidischolae sp . nov; Peltola JS et al.; Nocardiopsis strains were isolated from water-damaged indoor environments . Two strains (N . alba subsp . alba 704a and a strain representing a novel species, ES10.1) as well as strains of N . prasina, N . lucentensis, and N . tropica produced methanol-soluble toxins that paralyzed the motility of boar spermatozoa at <30 microg of crude extract (dry weight) x ml(-1) . N . prasina, N . lucentensis, N . tropica, and strain ES10.1 caused cessation of motility by dissipating the mitochondrial membrane potential, Deltapsi, of the boar spermatozoa . Indoor strain 704a produced a substance that destroyed cell membrane barrier function and depleted the sperm cells of ATP . Indoor strain 64/93 was antagonistic towards Corynebacterium renale . Two indoor Nocardiopsis strains were xerotolerant, and all five utilized a wide range of substrates . This combined with the production of toxic substances suggests good survival and potential hazard to human health in water-damaged indoor environments . Two new species, Nocardiopsis exhalans sp . nov . (ES10.1T) and Nocardiopsis umidischolae sp . nov . (66/93T), are proposed based on morphology, chemotaxonomic and physiological characters, phylogenetic analysis, and DNA-DNA reassociations. Appl Environ Microbiol, 2001 Sep, 67(9), 4206 - 14 DNA from uncultured organisms as a source of 2,5-diketo-D-gluconic acid reductases; Eschenfeldt WH et al.; Total DNA of a population of uncultured organisms was extracted from soil samples, and by using PCR methods, the genes encoding two different 2,5-diketo-D-gluconic acid reductases (DKGRs) were recovered . Degenerate PCR primers based on published sequence information gave internal gene fragments homologous to known DKGRs . Nested primers specific for the internal fragments were combined with random primers to amplify flanking gene fragments from the environmental DNA, and two hypothetical full-length genes were predicted from the combined sequences . Based on these predictions, specific primers were used to amplify the two complete genes in single PCRs . These genes were cloned and expressed in Escherichia coli . The purified gene products catalyzed the reduction of 2,5-diketo-D-gluconic acid to 2-keto-L-gulonic acid . Compared to previously described DKGRs isolated from Corynebacterium spp., these environmental reductases possessed some valuable properties . Both exhibited greater than 20-fold-higher kcat/Km values than those previously determined, primarily as a result of better binding of substrate . The Km values for the two new reductases were 57 and 67 microM, versus 2 and 13 mM for the Corynebacterium enzymes . Both environmental DKGRs accepted NADH as well as NADPH as a cosubstrate; other DKGRs and most related aldo-keto reductases use only NADPH . In addition, one of the new reductases was more thermostable than known DKGRs. J Endotoxin Res, 2001, 7(1), 25 - 33 Cloning, sequencing, and functional analysis of three glycosyltransferases involved in the biosynthesis of the inner core region of Klebsiella pneumoniae lipopolysaccharide; Noah C et al.; The genes encoding the 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) transferase (waaA) and heptosyltransferases I (waaC) and II (waaF) in Klebsiella pneumoniae were cloned from a DNA library by functional complementation of corresponding Escherichia coli and Salmonella enterica mutants . Sequence analyses revealed extensive homologies of the deduced proteins to their counterparts in other Enterobacteriaceae . However, differences were evident with regard to the chromosomal organization of the genes . To perform in vitro studies, the waaA, waaC and waaF genes were subcloned and expressed in the Gram-positive host Corynebacterium glutamicum . WaaA was characterized as a bifunctional enzyme capable of transferring two Kdo residues to a synthetic bisphosphorylated tetraacyl-lipid A precursor of E . coli (compound 406) . In contrast, waaC and waaF were shown to encode specific glycosyltransferases catalyzing the consecutive transfer of two L-glycero-D-manno-heptose residues to Kdo(2)-406. J Exp Bot, 2001 Sep, 52(362), 1785 - 803 Single and double overexpression of C(4)-cycle genes had differential effects on the pattern of endogenous enzymes, attenuation of photorespiration and on contents of UV protectants in transgenic potato and tobacco plants; Hausler RE et al.; To improve the efficiency of CO(2) fixation in C(3) photosynthesis, C(4)-cycle genes were overexpressed in potato and tobacco plants either individually or in combination . Overexpression of the phosphoenolpyruvate carboxylase (PEPC) gene (ppc) from Corynebacterium glutamicum (cppc) or from potato (stppc, deprived of the phosphorylation site) in potato resulted in a 3-6-fold induction of endogenous cytosolic NADP malic enzyme (ME) and an increase in the activities of NAD-ME (3-fold), NADP isocitrate dehydrogenase (ICDH), pyruvate kinase (PK), NADP glycerate-3-P dehydrogenase (NADP-GAPDH), and PEP phosphatase (PEPP) . In double transformants overexpressing cppc and chloroplastic NADP-ME from Flaveria pringlei (fpMe1), cytosolic NADP-ME was less induced and pleiotropic effects were diminished . There were no changes in enzyme pattern in single fpMe1 overexpressors . In cppc overexpressors of tobacco, the increase in endogenous cytosolic NADP-ME activity was small and changes in other enzymes were less pronounced . Determinations of the CO(2) compensation point (Gamma*) as well as temperature and oxygen effects on photosynthesis produced variational data suggesting that the desired decline in photorespiration occurred only under certain experimental conditions . Double transformants of potato (cppc/fpMe1) exhibited the most consistent attenuating effect on photorespiration . In contrast, photorespiration in tobacco plants appeared to be diminished most in single cppc overexpressors rather than in double transformants (cppc/fpMe1) . In tobacco, introduction of the PEP carboxykinase (PEPCK) gene from the bacterium Sinorhizobium meliloti (pck) had little effect on photosynthetic parameters in single (pck) and double transformants (cppc/pck) . In transgenic potato plants, increased PEPC activities resulted in a decline in UV protectants (flavonoids) in single cppc or stppc transformants, but not in double transformants (cppc/fpMe1) . PEP provision to the shikimate pathway inside the plastids, from which flavonoids derive, might be restricted only in single PEPC overexpressors. Kyobu Geka, 2001 Aug, 54(9), 753 - 7 {Bacterial contamination of salvaged blood in open heart surgery: is that an airborne contamination or a normal skin flora contamination?}; Ishida T et al.; We investigated sources of bacterial contamination of intraoperative salvaged blood producted by autologous transfusions device (CS; CELL SAVER 5, Heamonetics Corp., Braintree, MA) . Eleven patients undergone open heart surgeries including 2 emergency operations with a median sternotomy enrolled in this study . Blood samples were drawn from salvaged blood bags . Airborne contaminants (AB) were collected by a blood agar plate put besides the operation bed for 30 minutes . The median wounds samples were collected by a swab . Bacterial growth was detected in 81.8% of salvaged blood samples . Twenty-nine bacterium were isolated from CS, 72.4% of those were Staphylococci . 9.1% of sample was positive in wound swabs . Forty bacterium were isolated from plate cultures . 65% of them were Staphylococci . Staphylococcus epidermidis and coagulase negative Staphylococcus isolated both CS and AB in the 2 cases had the same identify codes, and incubated from several AB cultures . Corynebacterium sp . is also isolated from both CS and AB cultures in other 2 same cases . In 7 out of 8 cases (87.5%), from which Staphylococci isolated in CS, the Staphylococci were cultured from AB in not the same but the other cases . In conclusion, highly incidence of the identification in identical code of Staphylococci indicated that the main source of CS contamination was highly suspected to AB. Int J Antimicrob Agents, 2001 Aug, 18(2), 161 - 5 Antimicrobial activity of trifluoromethyl ketones and their synergism with promethazine; Kawase M et al.; The antimicrobial effects of 30 trifluoromethyl ketones {1-30} were studied on various representative bacteria . Of the ketones, 4,4,4-trifluoro-1-phenyl-1,3-butanedione {10}, 1,1,1-trifluoro-3-(4,5-dimethyloxazol-2-yl)-2-propanone {11} and 1-(2-benzoxazolyl)-3,3,3-trifluoro-2-propanone {18} were found to exhibit potent antibacterial activity against the Gram-positive Bacillus megaterium and Corynebacterium michiganese, but not against Gram-negative bacteria such as Pseudomonas aeruginosa and Serratia marcescens . Compounds 11 and 18 inhibited the Escherichia coli . Compound 18 was also effective against yeasts . The combination of promethazine with 18 was significantly synergistic against E . coli strains, especially the proton pump deficient mutant . The results suggest that membrane transporters are the target of trifluoromethyl ketones . The inhibition was more marked in the proton pump deficient E . coli mutant than in the wild type, which suggested that the antibacterial effect of trifluoromethyl ketones is partly prevented by the proton pump system. Vet Clin North Am Food Anim Pract, 2001 Jul, 17(2), 359 - 71, vii Caseous lymphadenitis in small ruminants; Williamson LH; Caseous lymphadenitis is a contagious bacterial disease that affects sheep and goats . It is characterized by abscess formation in the skin, internal and external lymph nodes, and internal organs . The causative agent is Corynebacterium pseudotuberculosis . The disease can become endemic in a herd or flock and is difficult to eradicate by virtue of its poor response to therapeutics, its ability to