Journal: Scientific pages of pulmonology

Article Title: Method for the Culture of Mouse Pulmonary Microvascular Endothelial Cells

doi:

Figure Lengend Snippet: Products and supplies for culture of mouse pulmonary microvascular endothelial cells.

Article Snippet: ECIS array , Applied BioPhysics , Troy, NY, USA , 8W1E PC.

Techniques: Modification, Saline

Journal: FEBS Open Bio

Article Title: Antimicrobial ceragenins inhibit biofilms and affect mammalian cell viability and migration in vitro

doi: 10.1002/2211-5463.12235

Figure Lengend Snippet: Effect of ceragenins on viability of HaCaT cells as measured via MTT assay (A). Treatments consist of LL‐37 (black), CSA‐13 (gray), CSA‐44 (dark blue), CSA‐90 (light blue), CSA‐192 (red), CSA‐131 (pink), CSA‐138 (dark green), and CSA‐142 (light green). Data points show viability as percent of control‐treated cells and are a mean of four independent experiments ± SEM. The levels of statistical significance of the treatment compared to the media control are as follows: ** P < 0.01; * P < 0.05 (two‐way ANOVA with Bonferroni post‐test). Representative ECIS responses are shown for growth factors (B). Treatments consist of control media (red solid), epidermal growth factor at 5 ng·mL −1 (brown dash), and transforming growth factor β1 at 2 ng·mL −1 (yellow dash). Ceragenin effect on ECIS cell migration (C). Data in the table show time to closure as percent of control‐treated cells and are a mean of five independent experiments ± SEM. The levels of statistical significance of the treatment compared to the media control are as follows: ** P < 0.01 (nonparametric Kruskal–Wallis with Steel post‐test). Representative ECIS responses are shown for test conditions (D). Compared to control (red solid), stated treatments are at doses of 10 ng·mL −1 (blue dash), 50 ng·mL −1 (purple dash), 100 ng·mL −1 (black dash) for all ceragenins and additionally 1 μg·mL −1 (green dash) for CSA‐142.

Article Snippet: All experiments were performed on 96W1E+ ECIS arrays (Applied Biophysics).

Techniques: MTT Assay, Control, Migration

Journal: Journal of Intensive Care

Article Title: Leukadherin-1 ameliorates endothelial barrier damage mediated by neutrophils from critically ill patients

doi: 10.1186/s40560-018-0289-5

Figure Lengend Snippet: Endothelial barrier disruption by neutrophils isolated from healthy ( n = 14), trauma ( n = 8), and septic donors ( n = 6) and protection by LA-1. Change in normalized resistance with respect to time. T = 0 represents time of neutrophil addition to HUVECs activated by TNF for 4 h. a Representative tracing of electrical resistance across activated HUVEC monolayers in the presence of neutrophils obtained from healthy, trauma, or septic donors. b Values are mean ± SEM of normalized resistance at 60 min following PMN addition. Analyses were performed evaluating the effect of neutrophils obtained from critically ill trauma patients ( n = 8), from critically ill septic patients ( n = 6), and from healthy controls ( n = 14) and assayed in duplicate on each experimental day. c Representative ECIS tracing of neutrophils added to HUVEC monolayer 30 min after fMLP (10 −6 M). d Values are mean ± SEM of normalized resistance at 60 min following PMN addition ± fMLP. e – g Representative tracings of neutrophils ± fMLP activation in the presence or absence of LA-1. e Neutrophils obtained from healthy donors. f Neutrophils obtained from trauma patients. g Neutrophils obtained from septic patients. h Values are mean ± SEM of normalized resistance at 60 min following PMN addition ± fMLP. * p < 0.05 vs. healthy unstimulated; ** p < 0.05 vs. healthy fMLP-stimulated; # p < 0.05 vs. trauma unstimulated; ## p < 0.05 vs. trauma fMLP-stimulated; @ p < 0.05 vs. septic unstimulated; and @@ p < 0.05 vs. septic fMLP-stimulated. Statistical significance determined ( p < 0.05) using three-way ANOVA with post hoc Tukey analysis

Article Snippet: Electric cell-substrate impedance sensing (ECIS) cultureware electrode arrays (8W10E+) and 16-well array station were obtained from Applied BioPhysics (Troy, NY).

Techniques: Disruption, Isolation, Activation Assay

Journal: PLoS Pathogens

Article Title: Endothelial Cell Permeability during Hantavirus Infection Involves Factor XII-Dependent Increased Activation of the Kallikrein-Kinin System

doi: 10.1371/journal.ppat.1003470

Figure Lengend Snippet: Mock-, HTNV-, or ANDV-infected HUVEC were trypsinized, seeded onto ECIS chamberslides, and cultured until confluent. Media were removed and replaced with phenol red free EBM containing Zn 2+ . After an equilibration, 0.2 ml of phenol red free EBM containing Zn 2+ , FXII, PK, and HK (100 nM each) were added to cells (time zero) (black lines) (A). To measure inhibition of activation, some samples were treated with CTI (1 µM) (blue line), PKSI-527 (5 µM) (red line), or HOE 140 (1 µM) (green line) simultaneously with factors (B, C, & D). Real time measurements frequency measurements were taken throughout the assay.

Article Snippet: Arrays were then loaded into the ECIS 16-well array station and stabilized using ECIS software v1.2.92.0 (Applied Biophysics).

Techniques: Infection, Cell Culture, Inhibition, Activation Assay