Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 1: ENDS aerosol exposure causes increased cell death in human lung epithelial cells The BEAS-2B cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 2: ENDS aerosol exposure causes increased cell death in mouse lung epithelial cells The MLE12 cell line was exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N-), 100μM Nicotine (Nicotine), HEPA filtered air (Air), and room temperature (Untreated) for 4 min. Cell death was analyzed 24h after treatment by flow cytometry using Caspase-3/7 stain and SYTOXTM permeability dye. Representative plots are shown in (A) and quantification show in (B-E). Necrotic cells are defined as SYTOXTM +, Caspase-3/7- (i) (B). Secondary necrotic cells are dual positive (ii) (C). Apoptotic cells are Caspase-3/7+, SYTOXTM- (iii) (D). All cells with activated Caspase-3/7 (E). Experiments were performed 3 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, &=p<0.05 compared to PG/VG N-, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol, Flow Cytometry, Staining, Permeability

Journal: American Journal of Respiratory Cell and Molecular Biology

Article Title: Electronic Nicotine Delivery System Aerosol-induced Cell Death and Dysfunction in Macrophages and Lung Epithelial Cells

doi: 10.1165/rcmb.2019-0200oc

Figure Lengend Snippet: Figure 3: One week of ENDS aerosol exposure causes decreased trans-epithelial resistance, dissociation of cells, and cell death Primary human bronchial epithelial cells were differentiated on transwell inserts into a pseudostratified epithelial layer. These cells were exposed to ENDS aerosol with (PG/VG N+) and without nicotine (PG/VG N- ), HEPA filtered air (Air), 100uM nicotine (Nicotine), and room temperature (Untreated) for 4 min. Trans- epithelial resistance was measured using a WPI EVOM TEER meter and STX3 electrode. Measurements were taken in triplicate and averaged. Background readings of empty wells as well as baseline measurements of each sample were used to calculate the change in resistance over the course of the treatment. Dissociated and dead cells were detected using trypan blue exclusion and counting on a hemocytometer. Treatments occurred once per day for 7 days. Experiments were performed 2 times with an n > 3 per group. *=p<0.05 compared to untreated, #=p<0.05 compared to air, %=p<0.05 compared to Nicotine. Data were analyzed using ANOVA followed by Tukey’s multiple comparisons test. Mean values are shown with SEM.

Article Snippet: Primary human bronchial epithelial cells (ATCC® PCS-300-010TM) were cultured on 24-well transwell inserts and grown in PneumaCultTM Medium system (Stemcell Technologies Inc.).

Techniques: Aerosol

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Schematic depicting the protocol for hCECs, hCSSCs, and hiNSCs seeding onto silk scaffolds.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques:

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Evaluating the effects of the microenvironment (elevated glucose or dynamic perfusion) and different chemical stimulants (capsaicin and sodium lauryl sulfate) on cell viability and gene and protein expression using the corneal tissue model. ( Abbreviations : Interleukin-1 beta (IL-1β), Substance P (SP), Calcitonin gene-related peptide (CGRP); Gene names : CRCP -accessory protein for CGRP, SCN- sodium channel, KERA - keratocan (cornea-specific proteoglycan), LUM - lumican (proteoglycan abundant in the stroma)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Expressing, Concentration Assay, Permeability

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Common secreted proteins related to nerve activation or tissue inflammation present in conditioned media isolated from corneal constructs. The relative concentration ranges of each protein detected in conditioned media in response to various chemical stimuli have been previously reported by our group ( Deardorff et al., 2018 ; Siran et al., 2018 ). ( Abbreviations : Substance P (SP), Calcitonin gene-related peptide (CGRP), Interleukin-1 beta (IL-1β), Tumor necrosis factor-alpha (TNF-α), Matrix metalloproteinase-9 (MMP-9)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Activation Assay, Isolation, Construct, Concentration Assay

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Phenotypic markers of each cell type present in the corneal construct. ( Abbreviations : Transient receptor potential cation channel subfamily V member 1 (TRPV1), Transient receptor potential cation channel subfamily A member 1 (TRPA1), Transient receptor potential cation channel subfamily M member 8 (TRPM8)).

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Marker

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Troubleshooting guide for construct assembly and characterization.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Cell Culture, Microscopy, Incubation, Sterility, Isolation, Lysis, Homogenization, Binding Assay

Journal: Current protocols in toxicology

Article Title: Assembly and Application of a 3D Human Corneal Tissue Model: A Step-by-Step Guide

doi: 10.1002/cptx.84

Figure Lengend Snippet: Relative timeframe for completion of each step in the assembly, stimulation, and characterization of the corneal tissue model. The amount of time may vary significantly depending on the user and the total number of constructs generated. These timeframes are based on 12 constructs per experiment.

Article Snippet: SUBCULTURING OF hCECs Materials Complete corneal epithelial media ( Refer to Reagents and Solutions Section ) Pipette T75 flask Primary human corneal epithelial cells (hCECs, ATCC, cat. no. ATCC® PCS-700-010TM) FNC Coating Mix® (Athena Environmental Sciences, cat. no. 0407) (Optional) 500 mL cell culture flask Seed and maintain hCECs 1.

Techniques: Construct, Generated, Incubation, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay, Imaging, Lysis, Protein Concentration, BIA-KA, Western Blot

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Structure and in vitro activities of BKI-1748 against N. caninum (Nc-β-gal) and T. gondii (Tg-β-gal). (A) molecular structure of BKI-1748, MW = molecular weight; EC 50 = concentration of half maximal proliferation inhibition; *toxicity levels against human foreskin fibroblasts (HFF) and zebrafish embryos were determined previously [22]. (B,C) Dose-response curves for Neospora and Toxoplasma tachyzoites grown in HFF.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: In Vitro, Molecular Weight, Concentration Assay, Inhibition

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Litter size, parasite burden, neonatal and postnatal mortality rates of N. caninum infected mice treated with BKI-1748.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: Infection