Journal: Research in veterinary science

Article Title: In vitro protective efficacy of Lithium chloride against Mycoplasma hyopneumoniae infection.

doi: 10.1016/j.rvsc.2016.03.013

Figure Lengend Snippet: Fig. 1. Antibacterial activity of LiCl against M. hyopneumoniae (1 × 106 CCU/ml) infected PK-cells (1 × 105 cells/well). Cells infected with M. hyopneumoniae, XLW-1 for 10 h and treated with LiCl for 12 h. (A) DNA extracted and quantified by qRT–PCR (log10 copies of DNA/well). Data presented as Means ± SD (n = 3). ⁎p b 0.008 and ⁎⁎p b 0.001 vs. control. (B) M. hyopneumoniae treated with LiCl (direct inactivation) before inoculating into cells. (C) Cells treated with LiCl then infected with M. hyopneumoniae, XLW-1 (pre-infection). DNA was extracted and quantified by qRT–PCR as above. A direct effect on bacteria was observed but not in cells. Data presented as Means ± SD (n = 3) ⁎p b 0.05 and p b 0.54 vs. control. (D) Effect of LiCl treatment was evaluated by IFA. Cells washed, fixed and probed with primary (p46 monoclonal antibody) and secondary-FITC conjugated antibodies. FITC- fluorescence was measured at OD 535. ⁎⁎P b 0.001 vs. control.

Article Snippet: We further infected cells (1 × 105 cells/well) in 96-well plates with M. hyopneumoniae (1 × 106 CFU/ml) at 37 °C for 10 h. Cells washed and inoculated LiCl (10–40 mM) for 12 h. Expression level of M. hyopneumoniae surface antigen (p46) was evaluated by IFA using p46 monoclonal antibody (our laboratory) and FITC-conjugated goat antimouse secondary antibodies (Boster Biological Technology Company, China).

Techniques: Activity Assay, Infection, Quantitative RT-PCR, Control, Bacteria

Journal: Research in veterinary science

Article Title: In vitro protective efficacy of Lithium chloride against Mycoplasma hyopneumoniae infection.

doi: 10.1016/j.rvsc.2016.03.013

Figure Lengend Snippet: Fig. 2. Identification of anti-apoptotic mechanism of LiCl against PK-15 cells (1 × 105 cells/well) infected with M. hyopneumoniae (1 × 106 CCU/ml). PK-15 cells were infected with M. hyopneumoniae, XLW-1 for 10 h and treated with or without LiCl for 12 h. (A) Phenotype of apoptosis and anti-apoptotic effect of LiCl were analyzed by AO/EB staining. +Ve denotes for untreated cells while −Ve denotes for neither infected nor treated cells. Most representative fields are shown. (B) Calculated apoptosis rate is provided. Data presented as Means ± SD (n = 3).⁎⁎P b 0.001 vs. control. (C) LiCl treatment inhibits production of NO and (D) superoxide anion induced by M. hyopneumoniae infection. NO production and superoxide anion were evaluated by Griess assay and superoxide anion assay respectively in PK-15 cells infected with M. hyopneumoniae and treated with LiCl. Data presented as mean ± SD (n = 3). ⁎P b 0.05 and ⁎⁎⁎P b 0.001 vs. control. (E) LiCl inhibited caspase-3 activity. Caspase-3 activity was measured in PK-15 cell lysate infected with ~1 × 106 CCU/ml M. hyopneumoniae for 10 h prior to LiCl treatment. LiCl inhibited caspase-3 activity in a dose-dependent manner. Data presented as Means ± SD (n = 3). ⁎⁎P b 0.01 vs. control.

Article Snippet: We further infected cells (1 × 105 cells/well) in 96-well plates with M. hyopneumoniae (1 × 106 CFU/ml) at 37 °C for 10 h. Cells washed and inoculated LiCl (10–40 mM) for 12 h. Expression level of M. hyopneumoniae surface antigen (p46) was evaluated by IFA using p46 monoclonal antibody (our laboratory) and FITC-conjugated goat antimouse secondary antibodies (Boster Biological Technology Company, China).

Techniques: Infection, Staining, Control, Griess Assay, Activity Assay