Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 1. Normalized protein expression (NPX) of plasma biomarkers in surviving [sTBI (S); n 5 6] and non- surviving sTBI patients [sTBI (NS); n 5 4] compared with matched healthy controls (CTR, n 5 10). Biomarkers shown are (A) ST2, (B) IL6, (C) TNNI3, (D) TNFR-2, (E) CHI3L1, and (F) EN-RAGE. Reference groups shown are ovarian cancer patients (OvCa, n 5 32) and dementia patients (mild cognitive impair- ment, MCI, n 5 18; Alzheimer disease, AD, n 5 12). Dotted lines represent group median, bars show interquartile range. Mann Whitney U test; *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant. For more discussion see text.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Expressing, Clinical Proteomics, MANN-WHITNEY

Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 3. Plasma concentration of ST2 and TNFR-2 in healthy controls (n 5 10) and sTBI patients (n 5 10), as measured by commercially available ELISAs. Mann Whitney U test; ****P < 0.0001. For more details see text.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Clinical Proteomics, Concentration Assay, MANN-WHITNEY

Journal: The journal of applied laboratory medicine

Article Title: Plasma Protein Profiling by Proximity Extension Assay Technology Reveals Novel Biomarkers of Traumatic Brain Injury-A Pilot Study.

doi: 10.1093/jalm/jfab004

Figure Lengend Snippet: Fig. 4. Comparison between commercially available proximity extension assays (Olink Proteomics) and ELISAs. Plasma ST2 was measured in (A) healthy controls (n 5 10); Spearman’s correlation (rs) 5 0.8909, P 5 0.0011, and (B) sTBI patients (n 5 10); rs 5 0.9758, P < 0.0001. Plasma TNFR-2 was measured in (A) healthy controls (n 5 10); rs 5 0.9030, P 5 0.0008, and (B) sTBI patients (n 5 10); rs 5 0.8667, P 5 0.0022. Black line shows line of best fit.

Article Snippet: All sTBI and matched control samples were analyzed in duplicate using the commercially available ELISA kits, DRT200 TNFR2 Quantikine kit and the DST200 ST2 Quantikine kit (R&D Systems), according to manufacturer’s instructions.

Techniques: Comparison, Clinical Proteomics

Journal: Frontiers in Microbiology

Article Title: Probing Clostridium difficile Infection in Complex Human Gut Cellular Models

doi: 10.3389/fmicb.2019.00879

Figure Lengend Snippet: Production of C. difficile spores, toxins, and host responses to infection. (A) Colony counts of spores recovered after heat treatment, and total cells in the cell-associated C. difficile fraction (infected cell lysates) in the 2D epithelial model. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05 as determined by two-way ANOVA. (B) ELISA for C. difficile toxins A and B shows increased toxin production after extended infection. Toxins were measured from medium obtained from the apical compartment containing uninfected cell layers incubated for 24 or 48 h (Control) or cells infected with C. difficile for 3, 6, 24, or 48 h. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05 as determined by two-way ANOVA. Gray line represents the sensitivity of the test at 0.5 ng/ml. (C) ELISA for human IL-8 indicates increased IL-8 production at 24 and 48 h p.i. IL-8 was measured in medium obtained from the basolateral compartment containing uninfected cell layers incubated for 3, 6, 24, or 48 h (Control) or cells infected with C. difficile for 3–48 h. Gray line represents the limit of detection at 32 pg/ml. Data shown are the mean of three independent experiments and error bars indicate SD, * p < 0.05, ** p < 0.01, and *** p < 0.001, **** p < 0.0001 as determined by one-way ANOVA with Tukey’s test for multiple comparison.

Article Snippet: IL-8 production was also determined by analysis of basolateral supernatants from the VDC using a human IL-8 ELISA kit (R&D systems, Minneapolis, MN, United States) following the manufacturer’s instruction.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Control, Comparison

Journal: Frontiers in Microbiology

Article Title: Probing Clostridium difficile Infection in Complex Human Gut Cellular Models

doi: 10.3389/fmicb.2019.00879

Figure Lengend Snippet: C. difficile spores and toxin production, and host response to infection in the 3D-VDC model. (A) Colony counts of spores and total cells in the host cell-associated C. difficile fraction (infected cell lysates). Data shown are the mean of three independent experiments and error bars indicate SD, ns, not significant as determined by two-way ANOVA. (B) Toxin A and B levels from apical compartment supernatants as determined by ELISA in the 3D model. Data shown are the mean of three independent experiments and error bars indicate SD, ns, not significant as determined by two-way ANOVA. Gray line represents the sensitivity of the test at 0.5 ng/ml. (C) Human IL-8 levels in supernatants from the basolateral compartments with uninfected cells incubated for 24 h or with cells infected with C. difficile for 3 and 24 h, as determined by ELISA. Gray line represents the limit of detection at 32 pg/ml. Data shown are the mean of three independent experiments and error bars indicate SD, ** p < 0.01, as determined by the one-way ANOVA with Tukey’s test for multiple comparison.

Article Snippet: IL-8 production was also determined by analysis of basolateral supernatants from the VDC using a human IL-8 ELISA kit (R&D systems, Minneapolis, MN, United States) following the manufacturer’s instruction.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Comparison

Journal: Cancer immunology research

Article Title: Antitumor effects of CAR T cells redirected to the EDB splice variant of fibronectin

doi: 10.1158/2326-6066.CIR-20-0280

Figure Lengend Snippet: (A) Scheme of retroviral vectors encoding the EDB-CAR, a 2A sequence, and truncated CD19 (tCD19); SH: short hinge; TM: transmembrane domain. (B-C) EDB-CAR expression was determined on T cells 7 days post-transduction by flow cytometric analysis. (B) Representative histogram (Black-filled line: NT T cells; Red-filled line: EDB-CAR transduced T cells) and (C) summary plot (n=7; Student’s t-test; ****p<0.0001). (D) NT and EDB-CAR T cells were incubated for 24 hours with increasing concentrations of rhFN-EDB-coated wells. IFNγ production in supernatants was determined by ELISA (n=4; two-way ANOVA; ***p<0.001; ****p<0.0001). (E) EDB expression of LM7, A673, A549, and U87 tumor cells, and two primary fibroblast cell lines (Fib 1, Fib 2) determined by RT-qPCR; dotted line represents the ΔCt score above which samples are considered positive. (F) NT and EDB-CAR T cells were incubated for 48 hours with EDB-positive tumor cells. IFNγ and IL2 production in supernatants was determined by ELISA (two-way ANOVA; ****p<0.0001). (G) Cytolytic activity of NT or EDB-CAR T cells at a E:T ratio of 4:1 against EDB-positive tumor cells (MTS assay; n=3; two-way ANOVA; *p<0.05; **p<0.01; ***p<0.001). (H) Cytolytic activity of NT or EDB-CAR T cells against primary fibroblasts (data of Fib 1 and Fib 2 was combined) at the indicated E:T ratios, with A549 serving as controls (n=3; two-way ANOVA; ****p<0.0001). (I) NT, EDB-CAR, or mutEDB-CAR T cells were incubated for 48 hours with U87 or U87FN−/− cells. IFNγ production in supernatants was determined by ELISA (n=3; two-way ANOVA; ****p<0.0001). (J) Cytolytic activity of NT, EDB-CAR, or mutEDB-CAR T cells against U87 or U87FN−/− cells (n=3; two-way ANOVA; ****p<0.0001). Mean+SEM is shown in panels.

Article Snippet: Cytokines were measured using human IFNγ and IL2 ELISA kits (R&D Systems).

Techniques: Retroviral, Sequencing, Expressing, Transduction, Incubation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Activity Assay, MTS Assay