Periostin peptide affects fibrosis in the myocardium.

<p>Periostin peptide affects fibrosis in the myocardium.</p

Adult cardiac fibroblast proliferation is moderately increased after periostin stimulation.

<p>(<b>a</b>) The effect of periostin on cell proliferation was quantified by BrdU incorporation using a colorimetric detection assay. At 0.5 and 1 μg/ml concentration, periostin treatment results in statistically significant increase in proliferation (p = 0.01 and 0.02 respectively). Culture with 10% FBS resulted in robust proliferation. (* P<0.05, ** P<0.01 to Control) (<b>b</b>) Adult cardiac fibroblasts with (0.5 μg/ml) or without stimulation by periostin.</p

Periostin peptide induces sustained functional improvements after MI.

<p>Periostin peptide induces sustained functional improvements after MI.</p

Periostin improves regional function.

<p>Analysis of regional function by strain analysis. (<b>a</b>) Example of division of mid-ventricular short-axis image into six segments to determine regional strain. (<b>b</b>) Representative example of segmental circumferential strain before initiation of therapy shows lack of contractility in infarcted anterior segment (yellow line, designated A). (<b>c</b>) Strain analysis after combination of infarcted segments (MA+MAS) and non-infarcted segments (MIS+MI+ML+MAL). Ctr., control hearts receiving gelfoam with buffer (<i>n</i> = 6); PN, hearts receiving gelfoam with periostin peptide (<i>n</i> = 7). Statistical comparison between control and periostin peptide groups by t-test (a–e, * <i>P</i><0.05), and two-way ANOVA.</p

Periostin peptide induces cardiomyocyte proliferation after MI.

<p>Confocal images of representative sections from infarcted hearts, one week (<b>a</b>) and three months (<b>b</b>) after MI in vehicle-treated and Periostin-treated animal groups. Frozen sections were stained with Aurora B, Ki67, phospho-histone-H3 (P-H3), and alpha-sarcomeric actin. Blue, DAPI-stained nuclei; red, Aurora B, Ki67, P-H3; green, alpha-sarcomeric actin. A global increase in proliferation expression markers was detected in the epicardial zone of the Periostin treated pigs, 1 week post-MI. Aurora B and P-H3 expression markers were also detected 3 months after periostin treatment.</p

Study design: 20 kg female Yorkshire pigs underwent experimental MI and received epicardial control gelfoam (Ctr.) or periostin peptide gelfoam (PN) 2 days later. PN (<i>n</i> = 7); Ctr (<i>n</i> = 6).

<p>Study design: 20 kg female Yorkshire pigs underwent experimental MI and received epicardial control gelfoam (Ctr.) or periostin peptide gelfoam (PN) 2 days later. PN (<i>n</i> = 7); Ctr (<i>n</i> = 6).</p

Periostin peptide-treated animals have smaller infarct scars and have myocardial strips.

<p>(<b>a, b</b>) Quantification of scar volume by delayed enhancement (DE) MRI imaging 12 weeks after implantation of periostin peptide or control gelfoam. Representative examples showing DE in pink (<b>a</b>) and quantification as percentage of LV myocardium show smaller scar areas in periostin peptide-treated animals (<b>b</b>). (<b>c, d</b>) Quantification of scar volume by tetrazolium chloride (TTC) staining. Representative slabs of the left ventricle (<b>c</b>) and quantification (<b>d</b>) show smaller scar areas in periostin peptide-treated animals; Scale bars, 1 cm. Myocardial tissue was only detectable in the scar area of treated animals (<b>e, f</b>). The heart to body weight ratio remained unaffected, (<b>g</b>), while the expansion index was increased in swine receiving periostin (<b>h</b>). Statistical significance tested with t-test (b, d). Scale bars, 5 mm (e), 1 cm (c). PN (<i>n</i> = 7); Ctr (<i>n</i> = 6).</p

Periostin peptide improves vascularization in the peri-infarct area.

<p>An increase in CD31 positive and vWF positive endothelial cells is detected 3 months post-periostin treatment in the peri-infarcted area. Quantification reveals a significantly increase capillary density compared to control animals (P<0.05). Blue, DAPI-stained nuclei; red, alpha-sarcomeric actin; green, CD31/von-Willebrand-factor (vWf).</p

Structured Inquiry-Based Learning: <i>Drosophila</i> GAL4 Enhancer Trap Characterization in an Undergraduate Laboratory Course

<div><p>We have developed and tested two linked but separable structured inquiry exercises using a set of <i>Drosophila melanogaster</i> GAL4 enhancer trap strains for an upper-level undergraduate laboratory methods course at Bucknell University. In the first, students learn to perform inverse PCR to identify the genomic location of the GAL4 insertion, using FlyBase to identify flanking sequences and the primary literature to synthesize current knowledge regarding the nearest gene. In the second, we cross each GAL4 strain to a UAS-CD8-GFP reporter strain, and students perform whole mount CNS dissection, immunohistochemistry, confocal imaging, and analysis of developmental expression patterns. We have found these exercises to be very effective in teaching the uses and limitations of PCR and antibody-based techniques as well as critical reading of the primary literature and scientific writing. Students appreciate the opportunity to apply what they learn by generating novel data of use to the wider research community.</p></div