Journal: PLoS ONE

Article Title: Pharmacologic Inhibition of CXCL10 in Combination with Anti-malarial Therapy Eliminates Mortality Associated with Murine Model of Cerebral Malaria

doi: 10.1371/journal.pone.0060898

Figure Lengend Snippet: The table displays biological processes or functions enhanced by Atorvastatin treatment identified by Ingenuity Pathway Analysis as significantly (p <0.05) associated with the dataset.

Article Snippet: Human brain vascular endothelial cells (HBVEC; Biowhittaker, Walkersville, MD) were cultured at 37°C with 5% CO 2 in Complete Serum-free Medium Kit (Cell Systems, Kirkland, WA) supplemented with 2% fetal bovine serum (FBS; ATCC, Manassas, VA) and 100 U/ml of streptomycin, 100 U/ml of penicillin (Gibco, Grand Island, NY) and were harvested and passaged at about 70–90% confluence.

Techniques:

Journal: PLoS ONE

Article Title: IL-22 Signaling Contributes to West Nile Encephalitis Pathogenesis

doi: 10.1371/journal.pone.0044153

Figure Lengend Snippet: a ) 20 PFU of WNV was inoculated intracranially. At day 5 p.i., brain viral loads (shown by WNV-E expression) and cytokine/chemokine or chemokine receptor transcripts were quantified by q-PCR and normalized with mouse actin. b–d, f ) Mice were infected with 200 PFU of WNV via s.c. footpad injection. b ) Expression of select chemokine receptors by blood cells was quantified by q-PCR using specific primers and probes, and normalized with beta actin. c ) The Cxcr2 expression levels of whole blood cells at day 4 p.i. were determined by immunostaining and flow cytometry. The results are expressed as mean fluorescence intensity (MFI) per cell. d ) The mRNA levels of cxcl1 and cxcl5 in WT and Il22 −/− brains were assessed by q-PCR. Each dot in the plot represents one mouse; * p <0.05; ** p <0.01. e ) Expression of CXCL1 and CXCL5 mRNA by human brain microvascular endothelial cells (HBVEC) treated with human recombinant IL-22. Bars: the mean of the results with standard deviation. ** p <0.01 ( n = 3). f ) Numbers of human PMNs that migrated out of the upper chamber (2×10 5 ) across the HBVEC monolayer down to the lower chamber over 18 hours in the presence of various concentrations of IL-22. Each dot represents the mean cell number from one well (counted 4 times). ** p <0.001. g ) Numbers of mouse PMNs that migrated out of the upper chamber (2×10 5 ) across the mouse brain vascular endothelial cell (MBVEC) monolayer down to the lower chamber. Each dot represents the mean cell number from one well (counted 4 times). * p <0.05. h ) Quantification of Cxcl1 levels of MBVECs treated as in g ) by ELISA. i ) Immunofluorescence staining of WNV-E (green), neutrophils (7/4, red) and DAPI (blue) in the brain cortex sections at day 5 p.i. Arrows point to co-staining. Micrographs were acquired using a laser scanning confocal microscope (20X images are shown and are representative of n = 5 mice per group). A higher magnification (63x) of confocal images of WT brain section is shown below (WNV, red; 7/4, green, TO-PRO3, blue). Arrow heads indicate typical polynuclear morphology of neutrophils. j ) FACS analyses of brain neutrophils (CD45Ly6G + ) of WT and Il22 −/− mice on day 6 p.i. Each dot represents 2 mice. * p <0.05.

Article Snippet: Primary human brain microvascular endothelial cells (HBVECs) (ScienCell, #1000) were cultured in formulated ECM medium (ScienCell, #1001) and treated with human recombinant IL-22 at a concentration of 200 ng/ml.

Techniques: Expressing, Infection, Injection, Immunostaining, Flow Cytometry, Fluorescence, Recombinant, Standard Deviation, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Microscopy

Journal: CNS Neuroscience & Therapeutics

Article Title: Glutathione Prevents Free Fatty Acids‐Induced Oxidative Stress and Apoptosis in Human Brain Vascular Endothelial Cells Through A kt Pathway

doi: 10.1111/cns.12068

Figure Lengend Snippet: Dose‐ and time‐course of free fatty acids (FFA) effect on NO content in human brain vascular endothelial cells (HBVECs). (A) Fluorescent micrographs of representative experiments show the dose and time course of NO production induced by FFA incubation (dose for 50, 100, and 200 μM, time for 24, 48, and 72 h, respectively); (B) Dose‐ and time‐course of FFA effect on NO content in HBVECs, as identified by using a fluorescent probe (DAF‐AM). Data are mean ± SD (following the same). *P < 0.05, **P < 0.001 versus control cells; # P < 0.05 versus FFA‐treated cells (200 μM for 24 h); (C) Dose‐ and time‐course of FFA effect on eNOS/NOS in HBVECs. *P < 0.05, **P < 0.001 versus control cells; # P < 0.05 versus FFA‐treated cells (200 μM for 24 h).

Article Snippet: Human brain vascular endothelial cells and trypsin were supplied by Sciencell Research Laboratories (San Diego, CA, USA).

Techniques: Incubation, Control

Journal: CNS Neuroscience & Therapeutics

Article Title: Glutathione Prevents Free Fatty Acids‐Induced Oxidative Stress and Apoptosis in Human Brain Vascular Endothelial Cells Through A kt Pathway

doi: 10.1111/cns.12068

Figure Lengend Snippet: Dose‐ and time‐course of free fatty acids (FFA) effect on phosphorylated Akt content in human brain vascular endothelial cells (HBVECs). (A) Dose‐course effect of FFA incubation for 48 h on phosphorylated active Akt content in HBVECs identified by western blot. *P < 0.05, **P < 0.001 versus control cells; #P < 0.05 versus FFA‐treated cells (50 μM for 48 h); (B) Time‐course effect of 200 μM FFA on phosphorylated active Akt content in HBVECs identified by western blot. *P < 0.05, **P < 0.001 versus control cells; #P < 0.05 versus FFA‐treated cells (200 μM for 24 h).

Article Snippet: Human brain vascular endothelial cells and trypsin were supplied by Sciencell Research Laboratories (San Diego, CA, USA).

Techniques: Incubation, Western Blot, Control