bal fluid proteins  (Biogen Inc)

Journal: Frontiers in physiology

Article Title: Differential modulation of lung aquaporins among other pathophysiological markers in acute (Cl 2 gas) and chronic (carbon nanoparticles, cigarette smoke) respiratory toxicity mouse models.

doi: 10.3389/fphys.2022.880815

Figure Lengend Snippet: FIGURE 3 Quantitative assessment of compromised lung integrity based on protein filtration [BAL protein content (A)] and expression of cell tight junction proteins [OCLN (B) and ZO-1 (C)] in mice subjected to different respiratory exposure regimens for CNT, CSE, CNT + CSE, and Cl2, as described in the Figure 1 legend. Transcriptional expression of the cell tight junction proteins occludin (OCLN) and zona-occludens-1 (ZO-1) was measured using gene-specific qRT-PCR. Statistical differences are represented by p-values.

Article Snippet: Total protein in bronchoalveolar lavage fluid BAL protein content was determined using DC protein estimation kit (BioRad, Hercules, CA) following the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR

Journal: Physiological Reports

Article Title: CFTR dysfunction increases endoglin and TGF‐ β signaling in airway epithelia

doi: 10.14814/phy2.13977

Figure Lengend Snippet: Increased endoglin in CF bronchoalveolar lavage fluid ( BAL ) fluid. Quantitative analysis by human CD 105 ELISA kit (Cat.# EK 0644, Boster Immuno‐Leader Biotechnology) indicates a fivefold increase in endoglin in severe CF ( n = 14) compared to non‐ CF samples (* P < 0.05, n = 5; A ). Immunohistochemistry on paraffin‐embedded lung sections shows increased ENG (brown staining) in CF lungs with severe disease as compared to non‐ CF (100x; primary Ab: Rabbit polyclonal anti‐human ENG , 1:50 dilution, Santa Cruz, Calibration bar = 250 μ m) (B).

Article Snippet: Quantitative analysis of ENG levels was measured in BAL fluid of CF and non‐CF subjects by following manufacturer's protocol using the human CD105 ELISA kit (Boster Biological Technology, Cat#EK0644).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining

Journal: PLoS ONE

Article Title: PI3K-AKT Signaling via Nrf2 Protects against Hyperoxia-Induced Acute Lung Injury, but Promotes Inflammation Post-Injury Independent of Nrf2 in Mice

doi: 10.1371/journal.pone.0129676

Figure Lengend Snippet: Mice were treated i . p . with LY294002 (LY) or DMSO (vehicle) 1 h prior to exposure and at 24 h intervals during hyperoxia exposure. Mice were exposed to 95% oxygen for 48 h at which time lungs were harvested. The BAL fluid was collected to measure both total cell counts and protein concentration, as markers of lung inflammation and lung injury (permeability), respectively. (A) Total protein in the BAL fluid of room air or hyperoxia-exposed mice treated with vehicle or LY. (B) Total cells in the BAL fluid of room air- or hyperoxia-exposed mice treated with vehicle or LY. (C) Total neutrophils and macrophages in the BAL fluid of vehicle- or LY-treated mice were exposed to room air or hyperoxia. BAL fluids (100 μl) were cytospun onto microslides and stained with Diff Quick stain kit. Differential cell counts were enumerated by counting a total number of 300 cells. The horizontal and vertical lines were plotted as median ± interquartile range for each group (n = 4–5). One-way ANOVA with Bonferroni corrections was performed for multiple group comparisons. * P < 0.05, hyperoxia vs room air; † P < 0.05, vehicle vs LY.

Article Snippet: BAL fluid protein concentration was measured by Bio-Rad protein assay reagent (Cat # 500–0006, Bio-Rad, Hercules, CA).

Techniques: Protein Concentration, Permeability, Staining, Diff-Quik

Journal: PLoS ONE

Article Title: PI3K-AKT Signaling via Nrf2 Protects against Hyperoxia-Induced Acute Lung Injury, but Promotes Inflammation Post-Injury Independent of Nrf2 in Mice

doi: 10.1371/journal.pone.0129676

Figure Lengend Snippet: Nrf2 –/– mice were treated with either vehicle or LY and subsequently exposed to hyperoxia or room air as described in . (A) Total protein in the BAL fluid of room air or hyperoxia exposed Nrf2 –/– mice treated with vehicle or LY. (B) Total cells in the BAL fluid of room air or hyperoxia exposed Nrf2 –/– mice treated with vehicle or LY. (C) Total neutrophils and macrophages in vehicle or LY-treated mice exposed to room air or hyperoxia. The horizontal and vertical lines were plotted as median ± interquartile range for each group (n = 4–5). One-way ANOVA with Bonferroni corrections was performed for multiple group comparisons. * P = 0.05, hyperoxia vs room air; † P = 0.05, vehicle vs LY treated group.

Article Snippet: BAL fluid protein concentration was measured by Bio-Rad protein assay reagent (Cat # 500–0006, Bio-Rad, Hercules, CA).

Techniques:

Journal: Science translational medicine

Article Title: Prevention of Alveolar Destruction and Airspace Enlargement in a Mouse Model of Pulmonary Lymphangioleiomyomatosis (LAM)

doi: 10.1126/scitranslmed.3003840

Figure Lengend Snippet: Increases in MMPs, inflammatory cells, and cytokines and decrease in alveolar elastin are associated with TSC2-null lung lesion growth. (A to C) MMP expression assessed in the cell-free supernatant of the BAL fluid at the indicated time points. A multiplex assay was performed by Searchlight technology (Aushon Biosystems). Data are means ± SE of n = 11 in each group. ***P < 0.001 for days 15 and 20 versus day 0 by ANOVA (Bonferroni-Dunn). (D) Schematic representation of elastin and collagen disposition in the alveolar neck. (E) Volumes of elastin (Eln) and collagen (Col) in alveoli of Control and TSC2-null lesion–carrying mice. Elastin and collagen were analyzed in alveoli necks of vehicle-injected (Contr) and TSC2-null cell–injected (TSC2-null) mice and expressed as percentage of the total volume of the alveolar neck. Data are means ± SE. **P < 0.01 for Control versus TSC2-null lesion–carrying mice by ANOVA (Bonferroni-Dunn). (F and G) Representative images of alveoli necks of vehicle-injected (Control) (F) and TSC2-null cell–injected (TSC2-null) (G) mice. Verhoeff's elastin stain and Picro-Ponceau counterstain were used to detect elastin (black) and collagen (red), respectively. Arrowheads, elastin-enriched areas. Scale bar, 10 μm. (H to L) Proinflammatory cytokine and chemokine expression assessed with a multiplex assay in the cell-free supernatant of BAL fluid from mice at the indicated time points after tumor inoculation. (M) Inflammatory cell counts assessed from the BAL collected at times as indicated after tumor cell inoculation. Data are means ± SE of n = 11 in each group. *P < 0.05; **P < 0.01; ***P < 0.001 versus day 0 by ANOVA (Bonferroni-Dunn).

Article Snippet: ELISA was performed with a Searchlight Protein Array multiplex of cell-free supernatant of the BAL fluids at Aushon Biosystems.

Techniques: Expressing, Multiplex Assay, Injection, Staining

Journal: Science translational medicine

Article Title: Prevention of Alveolar Destruction and Airspace Enlargement in a Mouse Model of Pulmonary Lymphangioleiomyomatosis (LAM)

doi: 10.1126/scitranslmed.3003840

Figure Lengend Snippet: Rapamycin plus simvastatin rescues animal survival, prevents lesion growth and lung destruction, and abrogates MMP induction. Mice injected with diluent (Control) or TSC2-null cells were treated with vehicle, rapamycin (Rapa), simvastatin (Simva), and rapamycin + simvastatin (Rapa + Simva) from day 3 after injection. (A) Weight of control (black) and TSC2-null cell–injected (red) mice were examined from day 3 (arrows) to day 20 of experiment. Data are means ± SE of n > 5 in each group. *P < 0.01 for Control versus TSC2-null cell–injected mice by ANOVA (Bonferroni-Dunn). Arrows, beginning of treatment. (B to D) H&E staining of murine lungs. Scale bar, 500 μm. (B) Lesion/lung ratio (C) and MAAA analysis (D) were performed at day 20 after injection. Data are means ± SE of n > 8 in each group. **P < 0.001 for Control versus TSC2-null cell–injected vehicle-treated mice and for Rapa, Simva, and Rapa + Simva versus vehicle for TSC2-null cell–injected mice by ANOVA (Bonferroni-Dunn). (E to G) Expression of MMP-2 (E), MMP-3 (F), and MMP-9 (G), assessed in the cell-free supernatant of BAL at day 20 by multiplex assay. Data are means ± SE of n > 6 in each group. **P < 0.001 for compound- versus vehicle-treated mice by ANOVA (Bonferroni-Dunn).

Article Snippet: ELISA was performed with a Searchlight Protein Array multiplex of cell-free supernatant of the BAL fluids at Aushon Biosystems.

Techniques: Injection, Staining, Expressing, Multiplex Assay