Cell surface marker expression in DmpCre/Ai9⁺ and DmpCre/Ai9⁻ populations of BOC cultures.

<p>Primary BOC isolated from Dmp1Cre/Ai9 mice were grown for ten days before analysis. Nontransgenic cells were utilized as controls to set the gate for tdTomato⁺ population (A, mean±SEM, n = 8). Expression of pan-hematopoietic marker CD45 was evaluated in the BOC population (B). Expression of CD105, Sca1, and endothelial marker CD31 was evaluated in the CD45⁻ BOC cells (C). Dot-plots from a representative experiment are shown. Percentages within gated populations represent mean±SEM of four biological replicates. Lineage marker expression was analyzed on at least 10⁴ gated cells. Gates were set in accord to the non-stained sample (not shown).</p

Dmp1Cre/Ai9⁺ cells dedifferentiate <i>in vivo.</i>

<p>Digested bone chips derived from Dmp1Cre/Ai9 mice were mixed with collagen type I and implanted subcutaneously in immunodeficient mice, as shown by the representative x-ray image (A<b>,</b> arrows). Five and twenty-eight days following implantation, the distribution of tdTomato⁺ cells was determined by histology (B).</p

BOC are labeled by Dmp1Cre/Ai9 but do not express Dmp1-GFP.

<p>Primary bone chip outgrowth cell (BOC) cultures were obtained by enzymatic digestion of long bones derived from Dmp1-GFP (A–B) and Dmp1Cre/Ai9 transgenic mice (C–D). After 3–5 days of culture, cells started to crawl out of the chips (B, D, F). Dmp1-GFP expressing osteocytes can be observed within the bone chips while no GFP expression was observed in BOC by epifluorescence (A). BOC from Dmp1Cre/Ai9 chips showed tdTomato fluorescence in a small proportion of cells (C). None of the outgrowth cells derived from Dmp1Cre negative/Ai9 bone chips express Dmp1Cre/Ai9 (E). Upper panel, epifluorescence; lower panel, phase contrast imaging. Images were taken at 10X magnification and are representative of at least 5 independent cultures.</p

Adipogenic potential of BOCs.

<p>Sorted Dmp1Cre/Ai9⁺ and Dmp1Cre/Ai9⁻ cells were cultured under basal conditions for 3 days (indicated as day 0), then treated with adipogenic medium. Unlike Dmp1Cre/Ai9⁺, Dmp1Cre/Ai9⁻ cells showed evidence of adipogenesis (A). Adipogenic differentiation was assessed by Oil Red O staining (B) and adiponectin and adipsin expression levels by real-time PCR analysis (C). Real-time PCR results are presented as mean±SEM of data normalized to Gapdh expression and pooled from two biological replicates. * P<0.05 compared to Dmp1Cre/Ai9⁺ day 9 as determined by two-tailed Student’s t-test. Statistical analysis was only performed where expression was detectable in both samples. ND, not detectable.</p

Osteogenic potential of BOCs.

<p>Following sorting, Dmp1Cre/Ai9⁺ and Dmp1Cre/Ai9⁻ populations were cultured under basal conditions overnight (indicated as day 0) before treatment with osteogenic medium. After 9 days of osteogenic induction, Dmp1Cre/Ai9⁺ cells underwent mineralization (A). Osteogenic differentiation was confirmed by von Kossa staining (B) and real-time PCR analysis of Bsp and Dmp1 mRNA expression (C). Real-time PCR results are presented as mean±SEM of data normalized to Gapdh expression and pooled from two biological replicates. * P<0.05 compared to Dmp1Cre/Ai9⁺ day 9 as determined by two-tailed Student’s t-test. Bsp, bone sialoprotein; Dmp1, dentin matrix protein 1; ND, not detectable.</p

Time lapse imaging of Dmp1Cre/Ai9 derived bone chips.

<p>Bone chips were imaged every 40 minutes starting at day three of culture. The movement of Dmp1Cre⁺/Ai9 cells within the bone chips and their expulsion onto the culture plate was observed (moving cell is indicated by the arrow). Representative images spanning a 14 h 20 minute period from the time points indicated are shown. Imaging was performed on three independent cultures.</p

Histology of Dmp1-GFP and Dmp1Cre/Ai9 bones.

<p>Detection of GFP expression in Dmp1-GFP derived histological sections (A–D). A specific GFP signal was localized in osteocytes (arrowheads) of cortical bone in calvaria (A) and femur (B) and in trabeculae (C). Detection of tdTomato expression in histological sections derived from Dmp1Cre/Ai9 transgenic mice (A’–D’). Cre-directed recombination was detected in osteocytes (arrowheads) and in the osteoblast layer (arrows) of cortical bone from calvaria (A’) and femur (B’) and trabecular bone (C’). tdTomato expression was also present in skeletal muscle (B’). Sections derived from GFP negative (D) and Dmp1Cre negative (D’) mice served as controls. Images were taken at 10X magnification and are representative of histology performed on 6 mice of each genotype. BM, bone marrow; LB, long bone; Ob’s, osteoblasts; Oc’s, osteocytes.</p

Campus virtuales : revista científica iberoamericana de tecnología educativa

Resumen basado en el de la publicaciónResumen en inglésSe describe la creación y contenidos de una plataforma de e-learning llamada AprendeBn para el sistema educativo panameño. Se comienza realizando un análisis de la problemática educativa en Panamá a partir de los informes del Ministerio de Educación (MEDUCA). Posteriormente se hace una evaluación comparativa de las principales plataformas de este tipo y se describen las características más relevantes de cada una de ella. Se pretende desarrollar una plataforma para Panamá que se enfoca en el estudiante y educador panameño de los niveles de premedia y media. En la plataforma se utiliza el formato, las mejores ideas y la experiencia de otras de similares características pero teniendo en cuenta los factores locales y los objetivos del sistema educativo panameño. Se persigue contar con un recurso tecnológico y social educativo, donde tanto estudiantes como educadores cuenten con contenidos actualizados de los programas oficialesES

Additional file 1: Figure S1. of Chemokine signals are crucial for enhanced homing and differentiation of circulating osteoclast progenitor cells

Increased frequency of osteoclast progenitor cells and subsets expressing chemokine receptors in peripheral blood and synovial fluid samples of patients with RA and PsA. a Proportion of OCPs, subsets expressing macrophage colony-stimulating factor receptor (CD115) and RANK in peripheral blood of CTRL subjects and patients with arthritis, and SF samples (SF RA, SF PsA), assessed by flow cytometry. b Concentrations of soluble RANKL in SF of arthritic patients, measured by ELISA. c Chemokine receptor expression on OCPs in peripheral blood of CTRL subjects and patients with arthritis, and SF of patients with arthritis, assessed by flow cytometry. Values are presented as medians (middle line), with boxes representing IQR, whiskers representing 1.5 times the IQR, and squares or circles representing outliers. Group-to-group comparisons were performed using a nonparametric Mann-Whitney U test, and p values <0.05 are shown for comparisons made between PsA and other groups. Previously shown p values for comparisons between RA and CTRL groups (Figs. 2 and 3) are not shown again for the sake of visual clarity. (TIF 1768 kb

Physiological characteristics of rats from high-serotonin (5HT) and low-5HT sublines.

<p>A. Indicators of 5HT homeostasis shown as “fold difference” between high- and low-5HT animals with 95% confidence intervals. Reference values were (mean±SD): a) for platelet serotonin level (PSL) 0.80±0.08 μg 5HT/mg platelet protein; b) for platelet serotonin uptake (PSU) 0.69±0.07 nmol 5HT/mg platelet protein/min. Rats were 2 months (PSU measurements) and 12 months (gut <i>Mao-A</i>, <i>Tph1</i> and <i>5HTT</i> expression) of age. PSL and gut 5HT turnover data are given for animals of 2 and 12 months of age. B. No difference in 5HT production and storage in the gut was observed between high- and low-5HT rat sublines. 5HT visualized by using immunohistochemistry was documented at 40× magnification and is depicted by black arrows. C-E. Physical characteristics of high-5HT and low-5HT animals (mean±SD). High-5HT animals are represented by black squares, low-5HT animals by open circles. C—body weight; D—femur length; E—hanging time in the string test (mean values from three 60-sec trials separated by 10-min intervals). Relative differences (%) are shown for subline*age interaction contrasts. P-values were adjusted for multiple comparisons (n = 6–15 rats/group). H-L indicates a difference between high-5HT and low-5HT animals. 12–2 indicates a difference between 12 months and 2 months old animals. Mao-A—monoamine oxidase A; Tph1 –tryptophan hydroxylase 1; 5HTT—serotonin transporter</p