PMC:4236617 / 14783-16429 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/4236617","sourcedb":"PMC","sourceid":"4236617","source_url":"http://www.ncbi.nlm.nih.gov/pmc/4236617","text":"Inhibition of F4-fimbriated ETEC adherence to IPEC-1 cells by L. amylovorus\nL. amylovorus cells were cultivated overnight, collected and washed twice with PBS. The F4+ ETEC strain was cultivated overnight in LB broth containing 10 μCi 3H-thymidine/ml for metabolic labeling. Labeled ETEC cells were collected and washed with PBS and the expression of F4 fimbriae was confirmed with the Fimbrex slide agglutination test kit (VLA Scientific, New Haw, UK). The inhibitory effects of the strains were tested in three different experimental set-ups: exclusion, competition and displacement. In each arrangement, 100 μl of L. amylovorus strains in DMEM/Ham’s F-12 [1:1] medium (A600 = 6) were added to IPEC-1 cells; in exclusion assays 1 hour before, in displacement assays 1 hour after, and in competition assays simultaneously with the addition of 100 μl of 3H –labeled ETEC in the same medium (A600 = 0.6). In the displacement assays, the unbound ETEC cells were removed by two washes with PBS before the addition of lactobacilli. The cells were further incubated for one hour at +37°C, 5% CO2, followed by five washes with PBS. Finally, the cells were lysed by adding 250 μl of 1% SDS in 0.1 M NaOH per well and by incubating overnight at +37°C, and the radioactivity of the lysed samples was measured by liquid scintillation counting. The proportion of adherent ETEC cells (%) in the presence or absence of the L. amylovorus strains was calculated as in the adhesion experiments, and the inhibition (%) was calculated according to the formula: [adherence (no La) – adherence (with La)] / adherence (no La) x 100%, where La indicates L. amylovorus.","divisions":[{"label":"Title","span":{"begin":0,"end":75}}],"tracks":[{"project":"MicrobeTaxon","denotations":[{"id":"T104","span":{"begin":866,"end":870},"obj":"562"},{"id":"T105","span":{"begin":944,"end":948},"obj":"562"},{"id":"T106","span":{"begin":1361,"end":1365},"obj":"562"},{"id":"T95","span":{"begin":62,"end":75},"obj":"1604"},{"id":"T96","span":{"begin":76,"end":89},"obj":"1604"},{"id":"T97","span":{"begin":617,"end":630},"obj":"1604"},{"id":"T98","span":{"begin":1410,"end":1423},"obj":"1604"},{"id":"T99","span":{"begin":1632,"end":1645},"obj":"1604"},{"id":"T100","span":{"begin":1014,"end":1026},"obj":"1578"},{"id":"T101","span":{"begin":28,"end":32},"obj":"562"},{"id":"T102","span":{"begin":168,"end":172},"obj":"562"},{"id":"T103","span":{"begin":283,"end":287},"obj":"562"}],"namespaces":[{"prefix":"_base","uri":"http://purl.bioontology.org/ontology/NCBITAXON/"}],"attributes":[{"subj":"T104","pred":"source","obj":"MicrobeTaxon"},{"subj":"T105","pred":"source","obj":"MicrobeTaxon"},{"subj":"T106","pred":"source","obj":"MicrobeTaxon"},{"subj":"T95","pred":"source","obj":"MicrobeTaxon"},{"subj":"T96","pred":"source","obj":"MicrobeTaxon"},{"subj":"T97","pred":"source","obj":"MicrobeTaxon"},{"subj":"T98","pred":"source","obj":"MicrobeTaxon"},{"subj":"T99","pred":"source","obj":"MicrobeTaxon"},{"subj":"T100","pred":"source","obj":"MicrobeTaxon"},{"subj":"T101","pred":"source","obj":"MicrobeTaxon"},{"subj":"T102","pred":"source","obj":"MicrobeTaxon"},{"subj":"T103","pred":"source","obj":"MicrobeTaxon"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"MicrobeTaxon","color":"#ec9593","default":true}]}]}}