Journal: Journal of Cell Science

Article Title: MicroRNA-mediated regulation of p21 and TASK1 cellular restriction factors enhances HIV-1 infection

doi: 10.1242/jcs.167817

Figure Lengend Snippet: The p21 protein is regulated by let-7. The p21 protein is regulated by let-7. Flow cytometric analysis of p21 expression following 72 hours of let-7c overexpression (OE) in the Jurkat cell line, (A) sample result and (B) quantification of three experiments. (C) Cell cycle is affected by let-7 and p21 levels. At 4 days following let-7c overexpression in the Jurkat cell line, anti FITC-BrdU and 7-AAD were used in order to determine the proportion of cells in each cell cycle phase. Analysis was performed using Flowing Software 2 (Cell Imaging Core, Turku Centre for Biotechnology, Finland). The S:G1 and G2:G1 ratios in cells treated with let-7c or control are presented. Data in B,C show the mean±s.e.m. ( n = 3); * P <0.05 (paired Student's t -test).

Article Snippet: Both parameters were measured using FITC BrdU Flow Kit (BD Biosciences, USA) according to the manufacturer's protocol.

Techniques: Expressing, Over Expression, Software, Imaging

Journal: Medicina

Article Title: Missed Down Syndrome Cases after First Trimester False-Negative Screening—Lessons to be Learned

doi: 10.3390/medicina56040199

Figure Lengend Snippet: The seven cases with DS and false-positive risk calculation after first-trimester ultrasound risk assessment.

Article Snippet: We conducted traditional first-trimester screening, using maternal age at the time of measurement, of standardized ultrasound parameters (NT, nasal bone, DV, tricuspid flow, and fetal heart rate) and MoM values for biochemical markers (free beta-hCG level and serum PAPP-A), using ASTRAIA software.

Techniques: Software

Journal: Frontiers in Immunology

Article Title: TRAIL-Receptor 4 Modulates γδ T Cell-Cytotoxicity Toward Cancer Cells

doi: 10.3389/fimmu.2019.02044

Figure Lengend Snippet: Decreased Vδ1 T cell-cytotoxicity against TRAIL-R4 knockdown Colo357 cells. Ten thousand control- and TRAIL-R (TR) 4 KD Colo357 cells were cocultured with Vδ1 T cell lines coexpressing a Vγ2, 3, or 4 chain (closed symbol in A ) or a Vγ8 chain (open symbol in A ) with an E/T ratio of 25:1 and a final concentration of 12.5 U/mL IL-2 for 4 h. Thereafter, (A) cytotoxicity was analyzed by Real-Time Cell Analyzer and (B) PGE2 release out of the supernatant by ELISA. Fold change in Cell Index (CI) was calculated using formula as follows: C I - F o l d c h a n g e = 1 - ( ( 1 - ( S C ) * ( 1 1 - M C ) ) ; S, CI value of the sample; C, vale of the medium control; M, Mean of CI value of Triton-X-100 sample. Black lines indicate mean of six independent experiments. Significances are shown as P -value; ** = P < 0.01 and *** = P < 0.001. (C) After culturing 10 4 Colo357 cells (green line) in complete medium for 30 h, impedance of these adherent tumor cells expressed as CI was measured in 5 min steps. The CI was normalized to 1 shortly before the addition of substances as follows: Triton-X-100 to induce maximal lysis (black line), medium (green line), 1 μg/mL PGE2 (light blue line), Vδ2 γδ T cell line (brown line) or Vδ2 γδ T cell line plus PGE2 (dark blue lines) with an E/T ratio of 25:1 in the presence of 12.5 IU/mL rIL-2. CI was then measured in 1 min steps over additional 26 h. The loss of tumor cell impedance and thus a decrease of CI correlated with lysis of tumor cells. The average of triplicates and standard deviation were calculated; one representative experiment. Several replications of the experiments using four different Vδ2 T cell lines and five different Vδ1 T cell lines of different donors in independent experiments were performed (right panel). The cytotoxicity of γδ T cell lines against the indicated tumor cells in the presence of medium or PGE2 was calculated 4 h after addition of γδ T cell lines. The percentage of specific lysis was calculated by comparing measured samples to control sample without effector cells and maximal lysis. Statistical analysis was performed by t- test. Significances are shown as P- value; * P < 0.05 and ** P < 0.01.

Article Snippet: PGE2 was measured by Prostaglandin E2 Parameter Assay Kit (#SKGE004B) and human granzyme B by a sensitive sandwich ELISA (both from R&D System) in duplicates following the procedures outlined by the manufacturer.

Techniques: Knockdown, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Lysis, Standard Deviation

Journal: Frontiers in Immunology

Article Title: TRAIL-Receptor 4 Modulates γδ T Cell-Cytotoxicity Toward Cancer Cells

doi: 10.3389/fimmu.2019.02044

Figure Lengend Snippet: Increased Vδ1 T cell-cytotoxicity against Colo357 cells by bsAb [HER2xCD3] is further enhanced by COX inhibitors. Ten thousand control- or TRAIL-R (TR) 4 KD Colo357 cells were incubated for 24 h in 96-well E-plates in medium. After 23 h, a final concentration of 1 μg/mL bsAb [HER2xCD3] or 50 μM Indomethacin alone or in combination or alternatively in combination with 30 μM DuP697 or 2.5 mM valeryl salicylate as indicated were added. After 24 h, Vδ1 T cell lines of six different donors with an E/T ratio of 25:1 and a final concentration of 12.5 U/mL IL-2 were cocultured. (A) The release of granzyme B or (B) PGE2 were measured after 24 h by ELISA and (C) Vδ1 T cell-mediated cytotoxicity against Colo357 cells by RTCA shown as decrease in Cell Index (CI) fold change calculated with the formula mentioned in . Black bars represent mean of experiments. Significances are shown as P -value; ** = P < 0.01.

Article Snippet: PGE2 was measured by Prostaglandin E2 Parameter Assay Kit (#SKGE004B) and human granzyme B by a sensitive sandwich ELISA (both from R&D System) in duplicates following the procedures outlined by the manufacturer.

Techniques: Control, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: TRAIL-Receptor 4 Modulates γδ T Cell-Cytotoxicity Toward Cancer Cells

doi: 10.3389/fimmu.2019.02044

Figure Lengend Snippet: Enhanced PGE2 release and COX-expression of TRAIL-R4 knockdown Colo357 cells in comparison to control cells. (A) PGE2 release was determined in the supernatant of control KD–and two different TRAIL-R (TR) 4 KD (a) or (b)—Colo357 cells after overnight culture. (B,C) In parallel, cells were intracellularly stained with anti-COX-1 and anti-COX-2 mAb and analyzed by flow cytometry. The expression is shown as fold change in median fluorescence intensity (MFI) relative to isotype control. Bars represent mean ± SD of three independent experiments. Significances are shown as P -value; * = P < 0.05 and ** = P < 0.01.

Article Snippet: PGE2 was measured by Prostaglandin E2 Parameter Assay Kit (#SKGE004B) and human granzyme B by a sensitive sandwich ELISA (both from R&D System) in duplicates following the procedures outlined by the manufacturer.

Techniques: Expressing, Knockdown, Comparison, Control, Staining, Flow Cytometry, Fluorescence