Journal: Journal of Immunology Research

Article Title: PM2.5 Causes Increased Bacterial Invasion by Affecting HBD1 Expression in the Lung

doi: 10.1155/2024/6622950

Figure Lengend Snippet: Expression of HBD1 at different conditions within the cell model. (a) ELISA was used to detect the levels of secretory HBD1 in the supernatants of HBE cell cultures in the control, PM2.5, PAO1, and PM2.5 + PAO1 groups ( n = 3). (b) RT–qPCR was performed to detect the mRNA levels of HBE in the control, PM2.5, PAO1, and PM2.5 + PAO1 groups of HBE cells ( n = 3). (c) Cell lysate smear plate colony count ( n = 3). (d) RT–qPCR was performed to detect the mRNA levels of A549 in the control, PM2.5, PAO1, and PM2.5 + PAO1 groups of A549 cells ( n = 3). (a, b, d) ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. control, &&& P < 0.001 PAO1 vs. PAO1 + PM2.5 (one-way ANOVA). (c) ∗∗∗ P < 0.001 vs. PAO1, &&& P < 0.001 PAO1 + PM2.5 vs. HBD1 + PAO1 + PM2.5 (one-way ANOVA).

Article Snippet: HBD1 (Cat. No. NBP2-34906-5 μ g, Novus Biologicals, LLC, USA) was dissolved in sterile water containing 0.02% acetic acid and 0.4% BSA to create a 1 mg/mL storage solution, and stored at −20°C.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control, Quantitative RT-PCR

Journal: Journal of Immunology Research

Article Title: PM2.5 Causes Increased Bacterial Invasion by Affecting HBD1 Expression in the Lung

doi: 10.1155/2024/6622950

Figure Lengend Snippet: Effect of PM2.5 on bacterial infestation. (a) The effect of PM2.5 on the bacterial invasiveness of PAO1-infested HBE was examined by the colony method. Different concentrations (0, 50, 100, 200 μ g/mL) of PM2.5 stimulated HBE cells for 24 hr, and then the configured PAO1 bacterial solution was added to the exposure model at different infection complexes (MOI = 1, 10, 20) ( n = 3). (b) The number of intracellular viable bacteria after PAO1 infection with HBE stimulated by different concentrations of PM2.5 (0/25/50/100 μ g/mL) ( n = 3). (c) The mice were modeled with gross lung and HE staining performance (c1–c4) in the order of control, PM2.5, PAO1, and PAO1 + PM2.5 groups ( n = 6). (d) For HE staining microscopic magnification × 40x observation (d1–d4) in order of control, PM2.5, PAO1, and PAO1 + PM2.5 groups ( n = 6). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. control; &&& P < 0.001 PAO1 + PM2.5 vs. PAO1; ### P < 0.001 PAO1 + PM2.5 vs. HBD1 (one-way ANOVA). (e) H&E staining of mouse lung tissue for lung injury scoring ( n = 6).

Article Snippet: HBD1 (Cat. No. NBP2-34906-5 μ g, Novus Biologicals, LLC, USA) was dissolved in sterile water containing 0.02% acetic acid and 0.4% BSA to create a 1 mg/mL storage solution, and stored at −20°C.

Techniques: Infection, Bacteria, Staining, Control

Journal: Journal of Immunology Research

Article Title: PM2.5 Causes Increased Bacterial Invasion by Affecting HBD1 Expression in the Lung

doi: 10.1155/2024/6622950

Figure Lengend Snippet: Mouse model and HBD1 expression. (a) IHC microscopic observation of mouse lung tissue HBD1, (a1–a4) in the order of control, PM2.5 group, PAO1 group, and PAO1 + PM2.5 group (scale bars = 100 µ m, n = 6). (b) IHC scoring of lung tissue in mice ( n = 6). (c) ELISA for secreted HBD1 in the supernatant of alveolar lavage fluid ( n = 6). (d) Detection of DEFB1 mRNA expression in lung tissue homogenates by RT–qPCR ( n = 6). (e) Flowchart of the animal model. (f) Survival curves for each group of mice. (g) Colony count of mouse lung grinding homogenate dilution coated plates ( n = 6). (b–d) ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. control, && P <0.01, &&& P < 0.001 PAO1 vs. PAO1 + PM2.5 (one-way ANOVA). (g) ∗∗∗ P < 0.001 PAO1 vs. PAO1 + PM2.5, &&& P < 0.001 PM2.5 + PAO1 vs. HBD1 + PM2.5 + PAO1 (one-way ANOVA).

Article Snippet: HBD1 (Cat. No. NBP2-34906-5 μ g, Novus Biologicals, LLC, USA) was dissolved in sterile water containing 0.02% acetic acid and 0.4% BSA to create a 1 mg/mL storage solution, and stored at −20°C.

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Animal Model

Journal: Journal of Immunology Research

Article Title: PM2.5 Causes Increased Bacterial Invasion by Affecting HBD1 Expression in the Lung

doi: 10.1155/2024/6622950

Figure Lengend Snippet: P -value after log-rank statistics for survival data for each group of mice ( n = 10, log-rank test).

Article Snippet: HBD1 (Cat. No. NBP2-34906-5 μ g, Novus Biologicals, LLC, USA) was dissolved in sterile water containing 0.02% acetic acid and 0.4% BSA to create a 1 mg/mL storage solution, and stored at −20°C.

Techniques: Mouse Assay

Journal: Journal of Immunology Research

Article Title: PM2.5 Causes Increased Bacterial Invasion by Affecting HBD1 Expression in the Lung

doi: 10.1155/2024/6622950

Figure Lengend Snippet: Mouse model and HBD1 expression under inhibitor action. (a) Flowchart of the animal model. (b) Survival curves in each group under the effect of inhibitors. (c) Detection of DEFB1 mRNA expression in lung tissue homogenates by RT–qPCR ( n = 6). (d) ELISA for secreted HBD1 in the supernatant of alveolar lavage fluid ( n = 6). (e) IHC scoring of lung tissue in mice ( n = 6). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. PM2.5 + PAO1 ( t test).

Article Snippet: HBD1 (Cat. No. NBP2-34906-5 μ g, Novus Biologicals, LLC, USA) was dissolved in sterile water containing 0.02% acetic acid and 0.4% BSA to create a 1 mg/mL storage solution, and stored at −20°C.

Techniques: Expressing, Animal Model, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Antimicrobial Protein and Peptide Concentrations and Activity in Human Breast Milk Consumed by Preterm Infants at Risk of Late-Onset Neonatal Sepsis

doi: 10.1371/journal.pone.0117038

Figure Lengend Snippet: Concentrations of LF (A); HBD1 (B); HBD2 (C); HD5 (D); LL-37 (E); and total protein (F), measured in day 7 (n = 88) and day 21 (n = 77) breast milk samples using ELISA or BCA assay for protein. All data are shown on a log scale with line at median. **p <0.01, ***p<0.001; comparing levels at 7 and 21 days postpartum using Wilcoxon signed-rank tests.

Article Snippet: Standard curves from serial dilutions of purified human LF (Aviva systems biology, San Diego, CA, United States), recombinant HBD1, HBD2, or LL-37 (Innovagen), or a control peptide from the active region of human HD5 (US Biological, Salem, MA, United States) were used to interpolate concentrations in samples using a five-parameter logistic (5PL) curve fit.

Techniques: Enzyme-linked Immunosorbent Assay, BIA-KA

Journal: PLoS ONE

Article Title: Antimicrobial Protein and Peptide Concentrations and Activity in Human Breast Milk Consumed by Preterm Infants at Risk of Late-Onset Neonatal Sepsis

doi: 10.1371/journal.pone.0117038

Figure Lengend Snippet: Results from two experiments show median log-transformed colony-forming units (±interquartile range; n = 4) on a log scale of remaining CFU/mL from an inoculum of ~1 × 10 6 of: (A) S. epidermidis ; (B) S. aureus ; (C) E. coli ; or (D) S. agalactiae after 4 h incubation in LBWF (diagonal lined pattern) spiked with either no AMP (F; n = 8 shows both controls combined, statistical tests were performed using values from relevant day only), or with the low (L), median (M) or high (H) concentration of LF, HBD1, HBD2 or HD5 detected in breast milk samples. C = cocktail of LF, HBD1, HBD2 and HD5 at median concentrations (horizontal lined pattern). Shaded area indicates range of starting inoculum. Symbols depict the groups where statistically significant comparisons were made on log-transformed data (level of significance indicated by multiple symbols; *p <0.05, **p<0.01, ***p<0.001) in ANOVA with Dunnett’s multiple comparison tests of all high, median and low spiked treatments to LBWF control (*), or in a t-test of median LF to cocktail of LF plus other molecules (†).

Article Snippet: Standard curves from serial dilutions of purified human LF (Aviva systems biology, San Diego, CA, United States), recombinant HBD1, HBD2, or LL-37 (Innovagen), or a control peptide from the active region of human HD5 (US Biological, Salem, MA, United States) were used to interpolate concentrations in samples using a five-parameter logistic (5PL) curve fit.

Techniques: Transformation Assay, Incubation, Concentration Assay