Journal: Allergy, asthma & immunology research

Article Title: Serum Amyloid A1: A Biomarker for Neutrophilic Airway Inflammation in Adult Asthmatic Patients.

doi: 10.4168/aair.2022.14.1.40

Figure Lengend Snippet: Fig. 1. The production of SAA1 from AECs and the effects of SAA1 on proinflammatory cytokine production. SAA1 proteins were produced from AECs (SAECs and A549, respectively) in response to various stimuli (A, B), and in the coculture with PBNs, PBEs (C, D). SAA1-induced release of IL-6, IL-8 and S100A9 from AECs (SAEC and A549, respectively) (E-K). Data are presented as means ± standard error of mean (n = 6). P values were calculated by using one-way analysis of variance with Bonferroni's post hoc test. SAA1 enhanced the expression of p-ERK and p-p38 MAPK (L) in AECs (SAECs and A549, respectively). IL, interleukin; PBEs, peripheral blood eosinophils; PBNs, peripheral blood neutrophils; dex, dexamethasone; S100A9, S100 calcium-binding protein A9; p-ERK, phospho-extracellular signal-regulated kinase; ERK, extracellular signal-regulated kinase; p-p38 MAPK, phospho-p38 mitogen-activated protein kinase; MAPK, mitogen-activated protein kinase; AECs, airway epithelial cells; SAEC, small airway epithelial cells. *P < 0.050, †P < 0.010, ‡P < 0.001, ns = not significant.

Article Snippet: Epithelial cell culture and stimulation Two kinds of AECs were used in this study, primary small airway epithelial cells (SAECs) and human AEC line (A549), both of which were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Produced, Expressing, Binding Assay

Journal: Frontiers in pharmacology

Article Title: The Anticancer Effects of the Pro-Apoptotic Benzofuran-Isatin Conjugate (5a) Are Associated With p53 Upregulation and Enhancement of Conventional Chemotherapeutic Drug Efficiency in Colorectal Cancer Cell Lines.

doi: 10.3389/fphar.2022.923398

Figure Lengend Snippet: FIGURE 7 | Compound 5a decreases HT29 and SW620 cell viability without affecting the viability of normal colon epithelial CCD 841 cells and enhances irinotecan (IRI), 5-fluorouracil (5-FU), and oxaliplatin (OXA) cytotoxic effects on HT29 and SW620 cells. (A) HT29, SW620, and the normal colon epithelial CCD 841 cell lines were exposed to different concentrations (5-10-20 μM) of Compound 5a for 24 h. Cell viability was measured by the MTT assay at 540 nm regarding the cellular metabolic activity. Bar graph showing the cell viability percentage and the data are expressed as mean ± SD (n = 3). ***p < 0.001 and ****p < 0.0001 vs. Control. Half-maximal inhibitory concentrations (IC50) of Compound 5a on HT29 and SW620 cell viability were determined. HT29 (B,C) and SW620 (D,E) cells were treated with different concentrations of the chemotherapeutic drugs IRI, 5-FU, and OXA for 24 h in the presence (C,E) or absence (B,D) of various concentrations (5-10-20 μM) of Compound 5a. Cell cytotoxicity was measured by the MTT assay at 540 nm. Bar graph showing the cell viability percentage and the data are expressed as mean ± SD (n = 3). Half- maximal inhibitory concentrations (IC50) of each chemotherapeutic drug on HT29 and SW620 cell viability were also determined.

Article Snippet: Human normal colon epithelial cell (CCD841 CoTr), colorectal adenocarcinoma HT29, and mCRC SW620 cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA, United States) and grown in a complete medium composed of DMEM supplemented with 10% heatinactivated fetal bovine serum (FBS), 100 μg/ml streptomycin, 100 IU/ml penicillin and 2 mmol/l L-glutamine.

Techniques: MTT Assay, Activity Assay, Control

Journal: Developmental Cell

Article Title: Cigarette Smoke Exposure and Inflammatory Signaling Increase the Expression of the SARS-CoV-2 Receptor ACE2 in the Respiratory Tract

doi: 10.1016/j.devcel.2020.05.012

Figure Lengend Snippet: Cigarette Smoke Increases the Expression of ACE2 in Mouse and Human Lungs (A) ACE2 expression in the lungs of mice that were sham-treated or that were exposed to diluted cigarette smoke for 2, 3, or 4 h a day (low, medium, and high smoke exposure, respectively). (B) A diagram showing the approximate locations of tissue samples used in this analysis. Tracheal, large airway epithelial, and small airway epithelial specimens were collected by fiberoptic bronchoscopy as described in their respective publications. Lung resections were collected surgically from various locations. (C) ACE2 expression in human tracheal epithelia analyzed according to smoking history. (D) ACE2 expression in human large airway epithelia analyzed according to smoking history. (E) ACE2 expression in human small airway epithelia analyzed according to smoking history. (F) A volcano plot comparing gene expression in the respiratory epithelia of current smokers and never smokers from (C)–(E). The dotted lines indicate various p value thresholds (e.g., genes located above the 10th percentile have a combined p value greater than 90% of the genes included in the meta-analysis). The location of ACE2 is indicated with a red star. (G) ACE2 expression in the lungs of a cohort of patients undergoing thoracic surgery analyzed according to the number of pack years each patient smoked. (H) ACE2 expression in the lungs of TCGA patients analyzed according to the number of pack years each patient smoked. (I) ACE2 expression in respiratory epithelia collected by fiberoptic bronchoscopy among either current smokers or former smokers. (J) A volcano plot comparing gene expression between current smokers and former smokers. The dotted lines indicate various p value thresholds (e.g., genes located above the 10 th percentile have a combined p value greater than 90% of the genes included in the meta-analysis). The location of ACE2 is indicated with a red star. Each panel displays log 2 -normalized ACE2 expression relative to a control group. Data analyzed in (A) were from GSE18344 . Data analyzed in (C) were from GSE13933 . Data analyzed in (D) were from GSE22047 . Data analyzed in (E) were from GSE64614 . Data analyzed in (G) were from GSE76925 . Data analyzed in (H) were from https://gdac.broadinstitute.org/ . Data analyzed in (I) were from GSE79209 . Additional information on the data sources and sample sizes are included in . ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005 (Student’s t test).

Article Snippet: Early-passage human tracheal epithelial cells and small airways epithelial cells were acquired from ATCC and cultured according to the supplier’s recommended method.

Techniques: Expressing, Gene Expression, Control

Journal: Developmental Cell

Article Title: Cigarette Smoke Exposure and Inflammatory Signaling Increase the Expression of the SARS-CoV-2 Receptor ACE2 in the Respiratory Tract

doi: 10.1016/j.devcel.2020.05.012

Figure Lengend Snippet: ACE2 Is Expressed in Secretory Club and Goblet Cells Along with Alveolar Type 2 Cells in the Mammalian Lung (A) T-SNE clustering of cells from the mouse lung. Cells expressing ACE2 are highlighted in the right panel. (B) Cells in the mouse lung that express various lineage markers (TMEM100 for endothelial cells, EPCAM for epithelial cells, PDGFRA for mesenchymal cells, and PTPRC for immune cells) are highlighted. (C) Cells expressing markers for various epithelial lineages are highlighted: RTKN2 for alveolar type 1 cells, LAMP3 for alveolar type 2 cells, FOXJ1 for ciliated cells, GABRP for both goblet and club cells, MUC5AC for goblet cells, and SCGB1A for club cells. (D) A track plot displaying the expression of ACE2 and several lineage-related genes in different cell populations obtained from Leiden clustering. (E) T-SNE clustering of cells from the human lung. Cells expressing ACE2 are highlighted in the right panel. (F) Cells in the human lung that express various lineage markers (TMEM100 for endothelial cells, EPCAM for epithelial cells, PDGFRA for mesenchymal cells, and PTPRC for immune cells) are highlighted. (G) Cells expressing markers for various epithelial lineages are highlighted: LAMP3 for alveolar type 2 cells, FOXJ1 for ciliated cells, TP63 for basal cells, GABRP for both goblet and club cells, MUC5AC for goblet cells, and SCGB1A1 for club cells. (H) A track plot displaying the expression of ACE2 and several lineage-related genes in different cell populations obtained from Leiden clustering. The gene expression data used in (A)–(D) are from GSE121611 . The gene expression data used in (E)–(H) are from GSE122960 . Additional information on the data sources and sample sizes are included in .

Article Snippet: Early-passage human tracheal epithelial cells and small airways epithelial cells were acquired from ATCC and cultured according to the supplier’s recommended method.

Techniques: Expressing, Gene Expression

Journal: Developmental Cell

Article Title: Cigarette Smoke Exposure and Inflammatory Signaling Increase the Expression of the SARS-CoV-2 Receptor ACE2 in the Respiratory Tract

doi: 10.1016/j.devcel.2020.05.012

Figure Lengend Snippet: Cigarette Smoke Causes the Expansion of ACE2 + Secretory Cells (A) T-SNE clustering of the transcriptomes from single cells derived from the airway epithelia of smokers and never smokers. Cells expressing ACE2 are highlighted in the right panel. (B) Cells in the human airway that express various lineage markers (FOXJ1 for ciliated cells, TP63 for basal cells, MUC5AC for goblet cells, and SCGB1A1 for club cells) are highlighted. (C) GO terms enriched among ACE2-correlated transcripts. (D) Dot plots displaying the expression of the top ten differentially expressed marker genes for various airway lineages and for ACE2+ cells. (E) The fraction of cells expressing the indicated marker genes are displayed. FOXJ1 is a marker for ciliated cells, MUC5AC is a marker for goblet cells, and TP63 is a marker for basal cells. (F) The fractions of cells found in each cell type cluster are displayed. (G) The number of counts per ACE2 + cell are displayed. (H) The fraction of ACE2 + cells co-expressing MUC5AC or that are found within the goblet/club cell cluster are displayed. (I) The 100 top-ranked differentially expressed genes from each cluster in (A) as well as the 100 genes most strongly correlated with ACE2 were used to re-analyze the bulk gene expression data from smokers and non-smokers in Figure 2 F. A volcano plot displays the mean expression change of each cell type signature. (J) The same transcriptional signatures as in Figure 4 I were used to re-analyze the data from current smokers and former smokers in Figure 2 J. A volcano plot displays the mean expression change of each cell type signature. (K) ACE2 expression in mouse tracheal explants undergoing mucociliary differentiation. (L) ACE2 expression in human airway epithelial cells undergoing mucociliary differentiation. (M) ACE2 expression in human airway epithelial cells that underwent mucociliary differentiation in the presence of clean air or cigarette smoke. (N) The same transcriptional signatures as in Figure 4 I were used to re-analyze the data from smoke exposure during differentiation from Figure 4 M. A volcano plot displaying the mean expression change of each cell type signature is displayed. Data analyzed in (A)–(H) were from GSE131391 . Data analyzed in (K) were from GSE75715 . Data analyzed in (L) were from GSE39059 . Data analyzed in (M) were from GSE135188 . Additional information on the data sources and sample sizes are included in . In (E), (F), and (H), a chi-square test is applied. In (G), a Mann-Whitney U test is applied. In (K)–(M), a Student’s t test is applied. ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005.

Article Snippet: Early-passage human tracheal epithelial cells and small airways epithelial cells were acquired from ATCC and cultured according to the supplier’s recommended method.

Techniques: Derivative Assay, Expressing, Marker, Gene Expression, MANN-WHITNEY

Journal: Developmental Cell

Article Title: Cigarette Smoke Exposure and Inflammatory Signaling Increase the Expression of the SARS-CoV-2 Receptor ACE2 in the Respiratory Tract

doi: 10.1016/j.devcel.2020.05.012

Figure Lengend Snippet: ACE2 Is an Interferon-Stimulated Gene that Is Upregulated by Viral Infections (A) ACE2 expression in airway epithelial cells that were infected with influenza. (B) ACE2 expression in airway epithelial cells that were infected with respiratory syncytial virus. (C) ACE2 expression in airway epithelial cells that were infected with SARS. (D) ACE2 expression in airway epithelial cells that were infected with MERS. (E) ACE2 expression in airway epithelial cells that were transfected with the dsRNA mimic poly(I:C). (F) ACE2 expression in human tracheal cells that were cultured in the presence of the indicated cytokine for 24 h. (G) ACE2 expression in human small airway epithelial cells that were cultured in the presence of the indicated cytokine for 24 h. (H) ACE2 expression in airway epithelial cells that were cultured in the presence of IFN-β. Each panel displays log 2 -normalized ACE2 expression relative to a control group. Data analyzed in (A) and (B) were from GSE32138 . Data analyzed in (C) were from GSE47963 . Data analyzed in (D) were from GSE100504 . Data analyzed in (E) were from GSE51392 . Data analyzed in (H) were from GSE19392 . Additional information on the data sources and sample sizes are included in . ∗ p < 0.05, ∗∗ p < 0.005, ∗∗∗ p < 0.0005 (Student’s t test).

Article Snippet: Early-passage human tracheal epithelial cells and small airways epithelial cells were acquired from ATCC and cultured according to the supplier’s recommended method.

Techniques: Expressing, Infection, Virus, Transfection, Cell Culture, Control