Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Proteins preferential to either HFpEF or control groups

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Control, Sequencing, Ubiquitin Proteomics

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Representative MS/MS scan for S100A8 peptide sequence ALNSIIDVYHK. Raw m/z spectral images with peak assignments and b and y ion lists along with a representation of peptide sequencing by tandem mass spectrometry

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Tandem Mass Spectroscopy, Sequencing, Mass Spectrometry

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Plasma levels of S100A8 in control vs. HFpEF groups. a S100A8 is found in increased levels in the plasma of subjects with HFpEF vs. control subjects as detected by ELISA. The MCW columns include the control (n = 7) and HFpEF (n = 9) from the discovery cohort and the NWU colums include the control (n = 18) and HFpEF (n = 25) samples from the validation cohort. *p < 0.006 vs MCW Control. # p < 0. 002 vs NWU Control

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: Overview of primary and secondary screening methods to identify potential mediators of HFpEF. a Platelet proteomes were subject to mass spectral analysis and novel proteins were identified. b Human cardiomyocytes derived from induced pluripotent stem cells were used to determine whether proteins that were identified in a had direct effects on cardiomyocytes function in vitro. Purified recombinant protein S100A8 was tested in this assay

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Derivative Assay, In Vitro, Purification, Recombinant

Journal: Journal of Translational Medicine

Article Title: Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction

doi: 10.1186/s12967-016-0774-3

Figure Lengend Snippet: S100A8-mediated effects on human iPSC-derived cardiomyocytes. a Shows example action potentials recorded from rS100A8 treated iPSC derived human cardiomyocytes. The addition of rS100A8 to the buffer extended the period between action potentials. This period is phase 4; the diastolic membrane potential between action potentials. b rS100A8 exacerbates the arrhythmic tendencies of human cardiomyocytes. c Spontaneous Ca 2+ transients recorded from human cardiomyocytes treated with rS100A8 as indicated by the blue line. rS100A8 significantly delayed the recovery of depolarization. Wash out of rS100A8 reversed these effects

Article Snippet: Recombinant S100A8 was purchased from Creative BioMart (Shirley, NY).

Techniques: Derivative Assay, Membrane

Journal: Cell Death & Disease

Article Title: USP18 is a key regulator of the interferon-driven gene network modulating pancreatic beta cell inflammation and apoptosis

doi: 10.1038/cddis.2012.158

Figure Lengend Snippet: Inhibition of USP18 exarcebates IFN α -, IFN γ - and PIC-induced cell apoptosis in INS-1E cells and primary rat beta cells. ( a , b ) INS-1E cells ( a ) or primary rat beta cells ( b ) were transfected with siCTRL or siUSP18. After 48 h of recovery, INS-1E cells were left untreated or treated with IFN α (1000 U/ml) or IFN γ (100 U/ml) for 24 h ( a ) and primary rat beta cells with IFN α for 48 h ( b ). Apoptosis was measured using HO/PI staining. Results are means±S.E.M. of 3–5 independent experiments; $$ P <0.01 and $$$ P < 0.001 versus untreated (i.e., not treated with cytokines) transfected with the same siRNA; *** P <0.001 versus siCTRL treated with the same cytokine; ANOVA. ( c ) INS-1E cells were transfected with siCTRL (C) or siUSP18 (U) and exposed to IFN α or IFN γ for 24 h. Expression of cleaved caspases-9 and -3 and α -tubulin (used as loading control) were measured by western blot. Results are representative of three independent experiments. ( d ) INS-1E cells were transfected as in ( a ). After 48 h of recovery, they were left untransfected or transfected with PIC for 24 h. Apoptosis was measured using HO/PI staining. Results are means±S.E.M. of three independent experiments; $$$ P < 0.001 versus untreated (i.e., not transfected with PIC); *** P <0.001 versus siCTRL transfected with PIC; ANOVA

Article Snippet: The following cytokine concentrations were used based on dose–response experiments previously performed by our group ( , ; unpublished data): recombinant rat IFN α (specific activity: 1 × 10 8 U/mg, PBL Biomedical Laboratories, Piscataway, NJ, USA) at 1000 U/ml, recombinant human IFN α at 2000 U/ml (specific activity: 1.8 × 10 8 U/mg, PeproTech, Rocky Hill, USA), recombinant rat IFN β at 1000 U/ml (specific activity: 8 × 10 7 U/mg, PBL Biomedical Laboratories), recombinant rat IFN γ at 100 U/ml (specific activity: 2 × 10 7 U/mg, R&D Systems, Abingdon, UK), recombinant human IL-1 β (specific activity 1.8 × 10 7 U/mg; a gift from C.W.

Techniques: Inhibition, Transfection, Staining, Expressing, Western Blot