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Pii: s0732-8893(02)00438-8

Diagnostic Microbiology and Infectious Disease Corynebacterium macginleyi isolation from conjunctival swab in Italy G.M. Giammancoa,*, V. Di Marcob, I. Priolob, A. Intrivicic, F. Grimontd, P.A.D. Grimontd aDipartimento di Igiene e Microbiologia “G. D’Alessandro”, Universita` di Palermo, Palermo, Italy bIstituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Palermo, Italy cAzienda Sanitaria Locale n.6, Palermo, Italy dUnite´ Biodiversite´ des Bacte´ries Pathoge`nes Emergentes, Institut Pasteur, Paris, France Received 29 April 2002; accepted 20 June 2002 Presented in part: Societa` Italiana di Microbiologia (SIM) 29th Congress, Genoa, Italy, November 2001.
Abstract
Corynebacterium macginleyi was isolated from conjunctival swabs of a farmer suffering from purulent conjunctivitis. This species has only recently been reported in Switzerland and Germany to be exclusively isolated from ocular surfaces. This represents the first isolationof C. macginleyi in Italy indicating that its circulation is not geographically limited. 2002 Elsevier Science Inc. All rights reserved.
1. Case report
colonies of Gramϩ pleomorphic rods were observed after48 h incubation on SBA. No growth was observed on The species Corynebacterium macginleyi has been re- chocolate agar and MacConkey agar. Gram’s stain revealed cently described by Riegel and coll. (Riegel et al., 1995) as coryneform rods forming arrangements similar to Chinese a result of comprehensive investigation of lipophilic coryne- letters. The commercial API Coryne system (bioMe´rieux, bacteria. Since the original description including three Marcy l’Etoile, France) was used according to the manu- strains isolated from eye specimens, only 25 clinical isolates facturer’s instructions for identification of the isolate. The of C. macginleyi have been reported in the literature. All of numerical code 5100305 was obtained after 24 h incubation these clinical strains have been isolated from conjunctival and used for species identification with API Coryne data- swabs in Switzerland (Funke et al., 1998) and Germany base 2.0 (Funke et al., 1997). API Coryne system unequivo- cously identified the isolate as C. macginleyi and the nu- We isolated C. macginleyi from a conjunctival swab of a merical code obtained corresponded to the one more 65 year old farmer living in a rural area in Western Sicily frequently encountered by Funke and coll. (Funke et al., and suffering from bilateral conjunctivitis. According to the 1998). Moreover, an identical identification code was ob- physician in charge, the patient showed conjunctival hype- tained with C. macginleyi type strain (ATCC 104099T).
raemia with a whitish discharge, loss of visual acuity and The Corynebacterium isolate was confirmed to be li- corneal ulcers, but with no signs of intraocular infection.
pophilic by testing its growth on Tween 80-supplemented The conjunctival sample was cultured on Columbia agar SBA (Riegel et al., 1995). Our isolate grew well on both 0.1 plates (Becton Dickinson BBL, Cockeysville, Md.) supple- mented with 5% sheep blood (SBA) and chocolate agar Antimicrobial susceptibility patterns were determined by (Becton Dickinson) at 37°C both in a 5% CO -enriched the agar diffusion method. Our strain was susceptible to a atmosphere and in ambient air and on MacConkey agar large panel of antibiotics including: amoxicillin/clavulanate, (Becton Dickinson) at 37°C in ambient air. Very small clindamycin, erythromycin, gentamicin, penicillin G, chlor-amphenicol, rifampicin, tetracycline, vancomycin, ce-foperazone, enrofloxacyn, and kanamycin.
In order to determine the phylogenetic position of our * Corresponding author. Tel.: ϩ39-0916553663; fax: ϩ39-091343896.
E-mail address: gmgiamm@libero.it (G.M. Giammanco).
isolate, 16S rRNA gene was amplified by polymerase chain 0732-8893/02/$ – see front matter 2002 Elsevier Science Inc. All rights reserved.
PII: S 0 7 3 2 - 8 8 9 3 ( 0 2 ) 0 0 4 3 8 - 8 G.M. Giammanco et al. / Diagnostic Microbiology and Infectious Disease 44 (2002) 205–207 Fig. 1. Phylogenetic dendrogram based on 16S rDNA sequences of reference strains of lipophilic species belonging to Corynebacterium genus and relativeposition of the first Italian C. macginleyi clinical isolate.
reaction and sequenced by GenomeExpress (Montreuil, C. macginleyi is reported to predominantly affect already France). The sequence of our clinical isolate was compared injured conjunctivas or superinfect during conjunctivitis and aligned to published 16S rRNA sequences searched caused by other bacterial micro-organisms or viruses with NCBI taxonomy browser (National Center for Biotech- (Funke et al., 1998; Joussen et al., 2000). No other bacteria nology Information, National Library of Medicine, Be- could be isolated from our sample but, unfortunately, it was thesda, MD, USA; http://www.ncbi.nlm.nih.gov) and re- not submitted for viral or Chlamydia isolation, so that bac- trieved from GenBank. The retrieved 16S rRNA sequences terial superinfection of a viral or chlamydial etiology can belonged to nine reference strains of six different species of lipophilic Corynebacteria, including C. macginleyi type The patient recovered uneventfully three weeks after strain, and three strains of CDC Groups F-1 and G. The sampling following Colbiocin (chloramphenicol ϩ colistin EMBL/GenBank accession numbers for the 16S rRNA se- ϩ tetracycline) antibiotic drops treatment. According to the quences were as follows: C. accolens ATCC 4972T, literature, our isolate was susceptible to a large panel of X80500; C. afermentans afermentans CIP 103499T, antibiotics including chloramphenicol and tetracycline. De- X82054; C. afermentans lipophilum CIP 103500T, X82055; layed recovery could be explained by the scarce compliance C. bovis NCTC 3224T, X84444; C. jeikeium ATCC 43216, of our patient toward therapy. Other three members of the U87816; C. jeikeium ATCC 43217, U87815; C. jeikeium same family showed similar symptoms but they were not ATCC 43734, U87823; C. CDC Group F-1 CDC G5911, sampled because they had already started antibiotic treat- X81904; C. CDC Group F-1 G4330, X81905; C. CDC ment. They all recovered after antibiotic drops treatment Group G CDC G5840, X80498; C. urealyticum ATCC but, in the absence of isolation, the occurrence of a C. 43042T, X81913; C. macginleyi ATCC 104099T, X80499.
macginleyi epidemic remains a mere speculation.
Phylogenetic analysis included sequence alignment, cal- Our finding of a case of conjunctivitis associated to C. culation of percent sequence similarity, construction of a macginleyi isolation in Italy confirms Joussen’s and coll.
phylogenetic tree, and assessment of the tree topology by indication that the presence of this micro-organism is not bootstrap analysis, which was performed by Clustal method geographically limited (Joussen et al., 2000).
with Weighted residue weight table using DNAstar software(DNASTAR Inc., Madison, WI, USA). In the phylogenetictree (Fig. 1) our clinical isolate clustered together with C. Acknowledgments
macginleyi published sequence showing 98.7% sequencesimilarity (0.4 divergence). 16S rDNA sequence similarity Thanks are due to G. Rubino for excellent technical above 97% should be considered sufficient to consider two strains as belonging to the same species (Stackebrandt &Goebel, 1994). The 16S rRNA gene sequence obtained forthe Italian C. macginleyi isolate has been deposited in theGenBank sequence data-base and given accession no.
References
As confirmed by 16S rDNA sequence analysis API Funke, G., Renaud, F. N. R., Freney, J., & Riegel, P. (1997). Multicenter Coryne system is affordable in identifying C. macginleyi evaluation of the updated and extended API (RAPID) Coryne database2.0. Journal of Clinical Microbiology, 35, 3122–3126.
isolates. Growth on Tween 80-supplemented SBA can be Funke, G., Pagano-Niederer, M., & Bernauer, W. (1998). Corynebacterium used to detect lipophilic isolates but further biochemical macginleyi has to date been isolated exclusively from conjunctival tests are needed for species identification.
swabs. Journal of Clinical Microbiology, 36, 3670 –3673.
G.M. Giammanco et al. / Diagnostic Microbiology and Infectious Disease 44 (2002) 205–207 Joussen, A. M., Funke, G., Joussen, F., & Herbertz, G. (2000). Coryne- of Corynebacterium macginleyi sp. nov. International Journal of Sys- bacterium macginleyi: a conjunctiva specific pathogen. British Journal temic Bacteriology, 45, 128 –133.
of Ophthalmology, 84, 1420 –1422.
Stackebrandt, E., & Goebel, B. M. (1994). Taxonomic note: a place for Riegel, P., Ruimy, R., de Briel, D., Prevost, G., Jehl, F., Christen, R., & DNA-DNA reassociation and 16S rRNA sequence analysis in the Monteil, H. (1995). Genomic diversity and phylogenetic relationships present species definition in bacteriology. International Journal of among lipid-requiring diphtheroids from humans and characterization Systemic Bacteriology, 44, 846 – 849.

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