Journal: Journal of Neuroinflammation

Article Title: FTY720 attenuates excitotoxicity and neuroinflammation

doi: 10.1186/s12974-015-0308-6

Figure Lengend Snippet: FTY720 prevents excitotoxic neuronal death in cerebrocortical primary and organotypic cultures. (A) Neuronal primary cultures 9 DIV were pre-treated with increased concentration of FTY720 (10 to 1,000 nM) for 24 h and then stimulated with N-methyl-D-aspartic acid (NMDA) (25 to 100 μM) in the same conditioned medium. After 24 h of NMDA treatment, lactate dehydrogenase (LDH) released in the medium was measured and mean ± SEM were reported as per cent of non-treated neurons ( n = 6, except for FTY720 1 μM, n = 2; two-way ANOVA: * or # P < 0.05; ** or ## P < 0.01; # vs control). (B) Organotypic cultures were pre-treated 24 h with FTY720 (10 and 100 nM) and then incubated with NMDA (50 μM) for 30 min, in the presence or in the absence of FTY720 or MK801 (50 μM, 10 min pre-incubation). FTY720 was added for additional 24 h. The damage was detected by PI uptake 24 h after treatment with NMDA and propidium iodide (PI) intensity was measured as corrected total fluorescence (CTF). Data are expressed as the mean ± SEM ( n ≥ 4 independent experiments; in each experiment, at least two slices for condition were used) and referred to percent of NMDA-treated neurons (dotted line). Student’s t test: * P < 0.05 and ** P < 0.01 vs NMDA-treated cells. (C) Representative photographs showing PI uptake in cortical slices treated as in (B). Scale bar: 1 mm.

Article Snippet: Cytotoxicity 96 colorimetric assay for the quantification of lactate dehydrogenase (LDH) release was purchased from Promega Corporation (Madison, USA).

Techniques: Concentration Assay, Control, Incubation, Fluorescence

Journal: Journal of Virology

Article Title: The Thr205 Phosphorylation Site within Respiratory Syncytial Virus Matrix (M) Protein Modulates M Oligomerization and Virus Production

doi: 10.1128/JVI.03856-13

Figure Lengend Snippet: CK2 activity enhances RSV infectious virus production. (A) A549 cells were treated for 48 h with 20 nM siRNA specific for CK2α or control siRNA, followed by infection with RSV at an MOI of 1 for 24 h. Cells were harvested, and cell-associated virus was analyzed by plaque assay on Vero cells. Results shown are represented as the percentages of plaque compared to that of the negative control and are the means ± standard errors of the means (SEM) (n = 3 experiments); significant difference (Student's t test) is indicated by the P value. (B) Vero or HEp-2 cells were infected with RSV at an MOI of 1 and subsequently treated with the CK2-specific inhibitor TBB at 2.8 ng/ml (24, 25) for 12 h either 6 or 18 h postinfection (p.i.). In the case of Vero cells, cell-associated virus was harvested at 30 h and 48 h p.i. and titrated on Vero cells. In the case of HEp-2 cells, culture supernatants and cell-associated virus were collected at 48 h p.i. and infectious virus titers were determined on Vero cells. Results for log TCID50/ml (mean ± SEM, n = 3 experiments) are shown as percentages relative to the control (no TBB). (C) Vero and HEp-2 cells were infected with RSV A2 at an MOI of 1. Cells were treated without or with 2.8 ng/ml TBB for 12 h at 6 to 18 h p.i. or 18 to 30 h p.i. The medium was then replaced with fresh medium without TBB, and cells were cultured further up to 48 h p.i. Cell toxicity was determined by LDH release in culture supernatants as described in Materials and Methods. Data (means ± standard deviations [SD]) from two independent experiments are represented as LDH activity, determined in triplicate, relative to no treatment.

Article Snippet: Release of lactate dehydrogenase (LDH) into the culture medium ( 28 ), as a measure of cell death, was estimated using the Promega nonradioactive LDH release assay as per the manufacturer's instructions.

Techniques: Activity Assay, Infection, Plaque Assay, Negative Control, Cell Culture

Journal: Proteome Science

Article Title: Proteomic analysis of effluents from perfused human heart for transplantation: identification of potential biomarkers for ischemic heart damage

doi: 10.1186/1477-5956-10-21

Figure Lengend Snippet: Part of the identified proteins that may be interesting in potential biomarker discovery of early ischemia in myocardium

Article Snippet: The concentration of three interested proteins identified by MS was quantitatively determinated with phosphoglycerate mutase 2(PGAM2) human ELISA kit (Cusabio biotech CO., LTD, China), L-lactate dehydrogenase B(LDHB) human ELISA kit and titin human ELISA kit (Uscn Life Science Inc, China) according to the manufacturer's instructions in a blinded manner.

Techniques: Biomarker Discovery, Marker, Transduction, Activity Assay, Variant Assay, Protein Binding, Protease Inhibitor, Binding Assay

Journal: Proteome Science

Article Title: Proteomic analysis of effluents from perfused human heart for transplantation: identification of potential biomarkers for ischemic heart damage

doi: 10.1186/1477-5956-10-21

Figure Lengend Snippet: Comfirmation of protein expression by ELISA . The concentration of three interesting proteins was quantitatively determinated with ELISA kits(n = 5). A . The mean concentration of LDHB. B . The mean concentration of PGAM2. C . The mean concentration of Titin. Value are expressed as means ± SEM for each group. # p < 0.05 represents a significant difference in the ischemia for 60 min group compared with the ischemia for 0 min group.

Article Snippet: The concentration of three interested proteins identified by MS was quantitatively determinated with phosphoglycerate mutase 2(PGAM2) human ELISA kit (Cusabio biotech CO., LTD, China), L-lactate dehydrogenase B(LDHB) human ELISA kit and titin human ELISA kit (Uscn Life Science Inc, China) according to the manufacturer's instructions in a blinded manner.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay