Expression of Robo4 in the fibrovascular membranes from patients with proliferative diabetic retinopathy and its role in RF/6A and RPE cells

Purpose: Robo4, a member of the roundabout (Robo) family, acts as a neuronal guidance receptor and plays some role in vasculogenesis and angiogenesis. This study investigated the effect of Robo4 on the formation of fibrovascular membranes (FVMs) from patients with proliferative diabetic retinopathy and its roles in choroid-retina endothelial (RF/6A) and human retinal pigment epithelial (RPE) cells. Methods: RT-PCR and immunohistochemistry were used to determine the levels of mRNA and the presence and distribution of Robo4 in FVMs. Small interfering RNA (siRNA) technology was used to knock down Robo4 expression and to study its effects on RF/6A and RPE cells in vitro. Cell proliferation, migration, spreading, cycling, and apoptosis were assessed with MTT assay, Boyden chamber assay, immunocytochemistry, and flow cytometry. Tube formation by RF/6A on Matrigel was also analyzed. Results: The level of Robo4 mRNA was high in FVMs. Robo4 was expressed in the vessels and fibrous-like tissue co-immunostained for CD31 and GFAP, respectively. Robo4 siRNA knockdown inhibited cell proliferation and migration. Tube formation by RF/6A cells was also disturbed. Under hypoxic conditions, more apoptotic cells were evident among the knockdown cells than among the control cells (p < 0.01). Conclusions: Robo4 may play a role in the formation of FVMs. Silencing the expression of Robo4 in RF/6A and RPE cells inhibited their proliferation and reduced their tolerance of hypoxic conditions, suggesting physiologic functions of Robo4 in the cells of the retina.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000267136400001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701Biochemistry & Molecular BiologyOphthalmologySCI(E)PubMed15ARTICLE112-131057-10691

RNAi-directed downregulation of OsBADH2 results in aroma (2-acetyl-1-pyrroline) production in rice (Oryza sativa L.)

<p>Abstract</p> <p>Background</p> <p>Aromatic rice is popular worldwide because of its characteristic fragrance. Genetic studies and physical fine mapping reveal that a candidate gene (<it>fgr</it>/<it>OsBADH2</it>) homologous to <it>betaine aldehyde dehydrogenase </it>is responsible for aroma metabolism in fragrant rice varieties, but the direct evidence demonstrating the functions of <it>OsBADH2 </it>is lacking. To elucidate the physiological roles of <it>OsBADH2</it>, sequencing approach and RNA interference (RNAi) technique were employed to analyze allelic variation and functions of <it>OsBADH2 </it>gene in aroma production. Semi-quantitative, real-time reverse transcription-polymerase chain reaction (RT-PCR), as well as gas chromatography-mass spectrometry (GC-MS) were conducted to determine the expression levels of <it>OsBADH2 </it>and the fragrant compound in wild type and transgenic <it>OsBADH2</it>-RNAi repression lines, respectively.</p> <p>Results</p> <p>The results showed that multiple mutations identical to <it>fgr </it>allele occur in the 13 fragrant rice accessions across China; <it>OsBADH2 </it>is expressed constitutively, with less expression abundance in mature roots; the disrupted <it>OsBADH2 </it>by RNA interference leads to significantly increased 2-acetyl-1-pyrroline production.</p> <p>Conclusion</p> <p>We have found that the altered expression levels of <it>OsBADH2 </it>gene influence aroma accumulation, and the prevalent aromatic allele probably has a single evolutionary origin.</p

Software defect management of electro hydraulic control system of hydraulic support

For software defect problem of electro hydraulic control system of hydraulic support, software structural features of the electro-hydraulic control system were analyzed from software testing method, software defect management framework was built, software testing schema was determined, and database structure of software defects database and software defect management flow were designed, so as to analyze software. The application shows that software quality and development efficiency of the electro-hydraulic control system are improved through software defect management

Crystal structure of (4aS,5S,6aS,6a1S, 10aS)-4a,5,6a,6a1,9,10-hexahydro-7H-4,5-methanocyclobuta[4,5]naphtho[8a,1-b]pyran-6(2H)-one, C15H16O2

C15H16O2, triclinic, P1̄ (no. 2), a = 5.6034(7) Å, b = 8.7105(11) Å, c = 12.5661(16) Å, α = 92.790(7)°, β = 100.516(7)°, γ = 106.105(6)°, V = 576.15(13) Å3, Z = 2, Rgt(F) = 0.0476, wRref(F2) = 0.1348, T = 296.15 K