Journal: PLoS pathogens

Article Title: Point mutations in FimH adhesin of Crohn's disease-associated adherent-invasive Escherichia coli enhance intestinal inflammatory response.

doi: 10.1371/journal.ppat.1003141

Figure Lengend Snippet: Figure 2. Adhesion ability of AIEC and non-AIEC strains with regard to fimH clade and CEACAM6 expression. (A) Cell-associated bacteria were quantified using non differentiated T84 cells after a 3 H infection period, and results were analyzed with regard to fimH clade. (B) Cell- associated AIEC bacteria belonging to the fimH S70/N78 clade were quantified using undifferentiated and differentiated T84 cells after a 3 H infection period. (C) Western blot analysis of whole protein extracts from non-differentiated and differentiated T84 cells using anti-CEACAM6 and anti-GAPDH antibodies. (D) Cell-associated bacteria (AIEC and non-AIEC bacteria belonging to the fimH S70/N78 clade) were quantified using differentiated T84 cells after a 3 H infection period. (E) Yeast agglutination titer of AIEC and non-AIEC strains belonging to the fimH S70/N78 clade. (F) Transmission electron micrograph of negatively stained AIEC and non-AIEC bacteria belonging to the fimH S70/N78 clade, magnification 625 000. doi:10.1371/journal.ppat.1003141.g002

Article Snippet: Adhesion assay of human intestinal and bladder epithelial cells T84 cells (derived from human colorectal carcinoma) and T24 cells (human bladder epithelial cells) were purchased from ATCC and maintained in an atmosphere containing 5% CO2 at 37uC in appropriate medium.

Techniques: Expressing, Bacteria, Infection, Western Blot, Agglutination, Transmission Assay, Staining

Journal: PLoS pathogens

Article Title: Point mutations in FimH adhesin of Crohn's disease-associated adherent-invasive Escherichia coli enhance intestinal inflammatory response.

doi: 10.1371/journal.ppat.1003141

Figure Lengend Snippet: Figure 4. Impact of FimH amino acid substitutions on AIEC adhesion ability. Cell-associated bacteria were quantified using undifferentiated (A) or differentiated (B) T84 cells after a 3 H infection period. WT represents the original AIEC LF82 reference strain and DfimH represents the LF82- DfimH isogenic mutant. The other constructs were made in the LF82-DfimH isogenic mutant, in which various fimH variants were reintroduced at the fimH locus. Each value is the mean 6 SEM of at least four separate experiments (* P,0.05; ** P,0.001; *** P,0.001). (C) Relative ability to bind Man1 (1 M/3 M binding ratio) with regard to undifferentiated T84 index adhesion of the various fimH chromosomal mutants constructed. (D) Bacterial binding to 1M-BSA under various shear stresses (pN/m cm2) was evaluated by measuring bacterial accumulation over 3 min. doi:10.1371/journal.ppat.1003141.g004

Article Snippet: Adhesion assay of human intestinal and bladder epithelial cells T84 cells (derived from human colorectal carcinoma) and T24 cells (human bladder epithelial cells) were purchased from ATCC and maintained in an atmosphere containing 5% CO2 at 37uC in appropriate medium.

Techniques: Bacteria, Infection, Mutagenesis, Construct, Binding Assay, Shear

Journal: Molecules

Article Title: Homobivalent Lamellarin-Like Schiff Bases: In Vitro Evaluation of Their Cancer Cell Cytotoxicity and Multitargeting Anti-Alzheimer’s Disease Potential

doi: 10.3390/molecules26020359

Figure Lengend Snippet: Cytotoxic activity of 5,6-dihydro-1-phenylpyrrolo[2,1- a ]isoquinoline (DHPPIQ) derivatives against a panel of tumor cell lines. a.

Article Snippet: Human cell cultures A549 (ATCC ® CCL-185TM, lung carcinoma), HCT116 (ATCC ® CCL-247TM, intestinal carcinoma), RD (ATCC ® CCL-136TM, rhabdomyosarcoma), SH-SY5Y (ATCC ® CCL-2266TM, neuroblastoma), HeLa (ATCC ® CCL-2TM, adenocarcinoma of the cervix uterus) and K562 (ATCC ® CCL243TM, chronic myelogenous leukemia) were maintained in Dulbecco’s modified Eagle’s medium (DMEM for A549, HCT116 and RD); DMEM/F12 1:1 (for SH-SH5Y); Eagle’s minimal essential medium (EMEM for HeLa) and Roswell Park Memorial Institute 1640 medium (RPMI 1640 for K562), with 10% fetal bovine serum, 2-mM l -glutamine and 1% gentamicin as an antibiotic at 37 °C under 5% CO 2 in a humid atmosphere.

Techniques: Activity Assay

Journal:

Article Title: Coordinate Intracellular Expression of Salmonella Genes Induced during Infection

doi:

Figure Lengend Snippet: Intracellular expression of ivi genes in cultured macrophages and cultured epithelial cells. Individual ivi fusions were incubated with cultured RAW 264.7 murine macrophages and cultured human epithelial cell lines HEp-2 (larynx carcinoma) and Henle-407 (embryonic small intestine). The coculture was incubated for 30 min, washed, treated with gentamicin to kill extracellular bacteria, washed, and incubated for 4 h. The cultured mammalian cells were lysed with Triton X-100, and β-galactosidase assays were performed on the recovered intracellular bacteria. For comparison, β-galactosidase assays were also performed on individual ivi fusions grown for 4 h in LB and in MEM supplemented with 10% FCS. Preselected Lac− and Lac+ strains were obtained from the initial nonselected pool of integrated IVET fusions. Error bars represent 1 standard deviation of the measured value.

Article Snippet: The murine macrophage cell line RAW 264.7 and human epithelial cell lines HEp-2 (larynx carcinoma) and Henle-407 (embryonic small intestine) (ATCC TIB-71, CCL-23, and CCL-6, respectively) were obtained from the American Type Culture Collection, Rockville, Md., and maintained in minimum essential medium (MEM) supplemented with Earle’s salts, l -glutamine, and 10% heat-inactivated fetal calf serum (FCS) (Life Technologies, Rockville, Md.).

Techniques: Expressing, Cell Culture, Incubation, Standard Deviation