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: Infection and Immunity

Article Title: Vaginal Epithelial Cell-Derived S100 Alarmins Induced by Candida albicans via Pattern Recognition Receptor Interactions Are Sufficient but Not Necessary for the Acute Neutrophil Response during Experimental Vaginal Candidiasis

doi: 10.1128/IAI.00861-13

Figure Lengend Snippet: In vivo PMN chemotactic ability of culture supernatants containing C. albicans-derived S100 alarmins. (A) PMN migration induced by S100 alarmins of C. albicans or E. coli origin. Overnight culture supernatants of S100A8, S100A9, and wild-type C. albicans were semisolidified with carboxymethylcellulose (3%) to reach a consistency of vaginal gel formulation. The vaginal gel preparations were administered intravaginally to estrogenized uninoculated mice once daily for 2 days in a volume of 20 μl per mouse using a microdispenser. Gel preparations of E. coli-derived recombinant S100A8 (rS100A8), S100A9 (rS100A9), or PBS (gel control) were tested in parallel. Vaginal lavage samples were collected prior to treatment and then daily after the last treatment. Vaginal PMNs were quantified by identifying PMNs by Pap smear staining and enumerating PMNs in 5 high-powered fields (×400 magnification) per mouse, and values were averaged. (B) Dose-dependent effects of S100A8-containing culture supernatant in vaginal PMN migration. Estrogenized uninoculated mice were treated with culture supernatant from S100A8-producing C. albicans at various dilutions. Vaginal lavage fluids were evaluated for PMNs at 48 h postadministration. The figure presents cumulative data from two repeats. *, P < 0.05 compared to the estrogenized untreated group (0 h). SEM, standard error of the mean.

Article Snippet: Recombinant mouse S100A8 and S100A9 ( Escherichia coli -derived; available by custom order from R&D Systems) or culture supernatants containing C. albicans -derived S100A8 or S100A9 were intravaginally administered to estrogenized mice.

Techniques: In Vivo, Derivative Assay, Migration, Formulation, Recombinant, Control, Staining

Journal: Infection and Immunity

Article Title: Vaginal Epithelial Cell-Derived S100 Alarmins Induced by Candida albicans via Pattern Recognition Receptor Interactions Are Sufficient but Not Necessary for the Acute Neutrophil Response during Experimental Vaginal Candidiasis

doi: 10.1128/IAI.00861-13

Figure Lengend Snippet: Antibody neutralization of S100A8 and S100A9 during infection. Estrogenized inoculated mice were intravaginally treated with anti-S100A8 (100 μg/ml), anti-S100A9 (100 μg/ml), or a combination of the two antibodies in a volume of 20 μl on days 1, 2, and 3 postinoculation. Vaginal lavage samples were collected prior to inoculation and on days 4 and 7 postinoculation. Vaginal PMNs were quantified by identifying PMNs by Pap smear staining and enumerating PMNs in 5 high-powered fields (magnification, ×400) per mouse and averaged. The figure presents cumulative data from three repeats with 5 mice per group. SEM, standard error of the mean.

Article Snippet: Recombinant mouse S100A8 and S100A9 ( Escherichia coli -derived; available by custom order from R&D Systems) or culture supernatants containing C. albicans -derived S100A8 or S100A9 were intravaginally administered to estrogenized mice.

Techniques: Neutralization, Infection, Staining

Journal: Infection and Immunity

Article Title: Vaginal Epithelial Cell-Derived S100 Alarmins Induced by Candida albicans via Pattern Recognition Receptor Interactions Are Sufficient but Not Necessary for the Acute Neutrophil Response during Experimental Vaginal Candidiasis

doi: 10.1128/IAI.00861-13

Figure Lengend Snippet: Role of S100 alarmins in the PMN response during infection. Quantification of vaginal Candida burden (A) and vaginal PMNs (B). The number of CFU per 100 μl of lavage fluids from estrogenized S100A8- and S100A9-deficient or C57BL/6 wild-type mice inoculated with C. albicans was assessed on day 4 postinoculation. PMNs in vaginal lavage fluid were identified by Pap smear staining and enumerated in 5 high-powered fields (magnification, ×400) per mouse. The figure presents cumulative data from three repeats with 10 mice per group. ***, P < 0.0001 compared to the wild-type control group. SEM, standard error of the mean.

Article Snippet: Recombinant mouse S100A8 and S100A9 ( Escherichia coli -derived; available by custom order from R&D Systems) or culture supernatants containing C. albicans -derived S100A8 or S100A9 were intravaginally administered to estrogenized mice.

Techniques: Infection, Staining, Control

Journal: Infection and Immunity

Article Title: Vaginal Epithelial Cell-Derived S100 Alarmins Induced by Candida albicans via Pattern Recognition Receptor Interactions Are Sufficient but Not Necessary for the Acute Neutrophil Response during Experimental Vaginal Candidiasis

doi: 10.1128/IAI.00861-13

Figure Lengend Snippet: In vitro screening of S100 alarmin response by PRR-deficient vaginal epithelial cells. Primary vaginal epithelial cell cultures derived from wild-type, MR−/−, TLR4-deficient, and SIGNR1−/− vaginal tissue explants were inoculated with C. albicans, and S100A8 (A) and S100A9 (B) mRNA expression was quantified by real-time PCR 24 h following inoculation. The expression of the target genes was normalized to that of the β-actin gene and expressed as the fold increase over expression in epithelial cells cultured alone in parallel. The figure presents cumulative data from two repeats. SEM, standard error of the mean.

Article Snippet: Recombinant mouse S100A8 and S100A9 ( Escherichia coli -derived; available by custom order from R&D Systems) or culture supernatants containing C. albicans -derived S100A8 or S100A9 were intravaginally administered to estrogenized mice.

Techniques: In Vitro, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Over Expression, Cell Culture