Measurement of dark-adapted electroretinography (ERG) amplitudes in the oxygen-induced retinopathy (OIR) model and normal mice.

<p>Amplitudes of a- (A) and b-waves (B) from the OIR model or from normal mice were measured at 4, 6, and 8 w. Stimulus flashes were used from −2.92 to 0.98 log cds/m². (C) Representative ERG waveforms at 4, 6, and 8 w. Values are expressed as the mean ± S.D., n = 4 to 6. ^*<i>P</i><0.05, ^**<i>P</i><0.01 versus Normal. OIR, oxygen-induced retinopathy model.</p

Retinal damage in the oxygen-induced retinopathy (OIR) model and normal mice.

<p>Retinal cross sections were prepared at 4 w and 8 w. (A) Hematoxylin and eosin staining. Scale bar, 50 µm. Retinal damage was evaluated by counting the number of cells in the GCL (B) and measuring the thickness of the IPL (C), INL (D), and ONL (E) in mice at 4 w and 8 w. Values are expressed as the mean ± S.D., n = 5 or 6. ^*<i>P</i><0.05, ^**<i>P</i><0.01 versus Normal. GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; ONL, outer nuclear layer; OIR, oxygen-induced retinopathy model.</p

Effects of fluoro-thalidomide on H929 cells.

<p>(A) Detection of cleaved PARP expression (after treatment with fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide, (<i>S</i>)-fluoro-thalidomide or cytosine β-D-arabinofuranoside at a concentration of 20 μg/mL for 24 h unless otherwise noted). (B) Detection of Fas (CD95) expression. (C) From left to right, detection of DNA fragmentation treated with cytosine β-D-arabinofuranoside, (<i>S</i>)-fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide, fluoro-thalidomide and untreated sample by agarose gel electrophoresis compared to DNA size marker (100bp DNA Ladder, Promega, Madison, USA). (D) Cell cycle analysis. The proportion of cells in the G1, S, G2/M or sub-G1 phase. Data are shown as mean (n = 3). (E) Phosphorylation expression of Bcl-2 Ser 70. Data are shown as mean ± SD (n = 3). ***, p < 0.001 **, p < 0.01 *, p < 0.05 vs. untreated.</p

Hematoxylin and eosin staining of retinal sections (thickness, 5 µm) obtained from mice at 7 days after the intravitreal injection of -methyl--aspartate (NMDA)

<p><b>Copyright information:</b></p><p>Taken from "Brazilian Green Propolis Protects against Retinal Damage and "</p><p>Evidence-based Complementary and Alternative Medicine 2006;3(1):71-77.</p><p>Published online Jan 2006</p><p>PMCID:PMC1375228.</p><p>© The Author (2006). Published by Oxford University Press. All rights reserved</p> Representative photographs showing non-treated normal retina (), NMDA-treated, vehicle-treated retina () and NMDA-treated, propolis-treated retina (). Propolis (PW; extract with water) at 100 mg kg (0.1 ml per 10 g) was intraperitoneally administered at 48 h, 24 h and 60 min before and at 6 h after the NMDA injection (40 nmol per eye). Treatment with propolis limited the damage to retinal ganglion cells (arrows) and to the IPL (vertical bars) induced by the NMDA injection. Also see . Vertical bars show thickness of plexiform layer. Horizontal bar represents 25 µm

The oscillatory potentials (OPs) amplitudes in response to a light flash in the oxygen-induced retinopathy (OIR) model and normal mice.

<p>The averaged OP amplitudes were measured at 4, 6, and 8 w. Values are expressed as the mean ± S.D., n = 4 to 6. ^*<i>P</i><0.05, ^**<i>P</i><0.01 versus Normal. OIR, oxygen-induced retinopathy model.</p

Effects of fluoro-thalidomide on H929, AGLCL and U937 cells.

<p>(A) Proportion of annexin V-positive cells in AGLCL cells, normal human B cells, by thalidomide, fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide or (<i>S</i>)-fluoro-thalidomide (Data are shown as mean ± SD (n = 3). ***, p < 0.001 **, p < 0.01 *, p < 0.05 vs. untreated, unless otherwise noted). (B) Proportion of annexin V-positive cells in U937 cells by thalidomide, fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide or (<i>S</i>)-fluoro-thalidomide. (C) MTT assay measuring (550 nm) the optical density of H929 cells treated with fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide, (<i>S</i>)-fluoro-thalidomide, or cytosine β-D-arabinofuranoside at a concentration of 20 μg/mL for 24 h. (D) Observation of morphological changes in H929 cells after treatment with fluoro-thalidomide, (<i>R</i>)-fluoro-thalidomide, (<i>S</i>)-fluoro-thalidomide, or cytosine β-D-arabinofuranoside at a concentration of 20 μg/mL for 24 h using Wright-Giemsa staining protocol. Apoptotic bodies (indicated by arrows) were observed after the treatment.</p

Fluoro-thalidomide not thalidomide promotes VEGF induced tube formation in HUVECs.

<p>(A) Representative photographs were shown in tube formation. Quantitative analysis of stained tube formation structures was measured by using angiogenesis imaging analyzer, version 2 in five different areas for each well. We measured joint (B), joints (B), paths (C), tube area (D), lengths (E). Data are shown as mean ± SEM (n = 3 or 4). ^##, p < 0.01 vs. Control group, and *, p < 0.05 **, p < 0.01 vs. Vehicle-treated group.</p

Structures of thalidomide (racemic form), fluoro-thalidomide (racemic form), and its (<i>R</i>)- and (<i>S</i>)- enantiomers, (<i>R</i>)-fluoro-thalidomide and (<i>S</i>)-fluoro-thalidomide.

<p>Structures of thalidomide (racemic form), fluoro-thalidomide (racemic form), and its (<i>R</i>)- and (<i>S</i>)- enantiomers, (<i>R</i>)-fluoro-thalidomide and (<i>S</i>)-fluoro-thalidomide.</p

BiP protein levels regulates retinal neovascularization.

<p>(A) HRMEC were cultured in a 96-well plate (at a density of 2×10³ cells/well), and were then supplemented with the indicated concentrations of BiX for 1 h, and measurements were made by WST-8 assay. Data are shown as mean ± S.E.M. (n = 6). *, p<0.05 vs. Control (Dunnett's multiple-comparison test). Migration of HRMEC was assessed using a wound-healing assay. (B) Images of the wounded monolayer of HRMEC were taken at 24 h after treatment for 1 h with BiX. (C) The indicated concentrations of BiX increased migration compared to the control. Scale bars indicate 500 µm. Data are shown as mean ± S.E.M. (n = 4). **, p<0.01 vs. Control (Dunnett's multiple-comparison test). The original images (D, F and J, L), together with the analyzed images (E, G and K, M) obtained using the Angiogenesis Tube Formation module in Metamorph, are shown. Scale bars indicate 500 µm and 100 µm (in box). Quantitative analysis was performed on the entire retinal microvasculature in flat-mounted retinas obtained at P17. BiX at 1 µM and BiP small interfering (si) RNA at 1 µg/mL increased (vs. vehicle) both the number of nodes (E, K) and the node areas (F, L), which are indexes of pathological neovascularization as calculated using the Angiogenesis Tube Formation module. Data are shown as mean ± S.E.M. (n = 4 or 5). *, p<0.05 vs.Vehicle (Student's <i>t</i>-test).</p

BiP was expressed in the retinal vascular endothelial cells of a murine oxygen-induced retinopathy (OIR) model.

<p>(A) OIR model mice at P17, treated with 3 µg/ml of tunicamycin at P14, were perfused with FITC-dextran and the retinas were stained with anti-BiP antibody. The original images (green channel) and BiP stained images (red channel) are shown, along with the analyzed images. (B) BiP (green channel) and CD31 (red channel) staining in flat-mounted retinas of P17 mice, under normoxia or subjected to the OIR protocol, following tunicamycin injection. Arrowheads and arrows indicate normal vessels and newly formed vessels anterior to internal limiting membrane, respectively. Scale bars indicate 50 µm. The complexes of BiP and T-cadherin in the retina were immunoprecipitated with the anti-T-cadherin antibody and detected by using Western blotting with the anti-BiP antibody (C), or by the Coomassie-staining (D).</p