Phosphoinositides are key regulators of diverse cellular processes. The pleckstrin homology (PH) domain mediates the action PtdIns(3,4)P2, PtdIns(4,5)P2 and PtdIns(3,4,5)P3, while FYVE relays pulse PtdIns3P. recent establishment that Phox (PX) interacts with PtdIns3P other phosphoinositides suggests another mechanism by which can regulate/integrate multiple events via a spectrum PX domain-containing proteins. Together discovery epsin N-terminal homologue (ENTH) PtdIns(4,5)P2, it is becoming...

Glaucoma is a disease frequently associated with elevated intraocular pressure that can be alleviated by filtration surgery. However, the post-operative subconjunctival scarring response which blocks efficiency major hurdle to achievement of long-term surgical success. Current application anti-proliferatives modulate not ideal as these often give rise sight-threatening complications. SPARC (secreted protein, acidic and rich in cysteine) matricellular protein involved extracellular matrix...

KIAA0305 is an uncharacterized member of the FYVE domain protein family. It closely related to SARA, with about 50% identity in carboxyl-terminal 800-amino acid region. Indirect immunofluorescence microscopy using polyclonal antibodies raised against revealed that it enriched early endosomes. The Myc-tagged version also faithfully targeted endosome. We have tentatively called endofin (forendosome-associated FYVE-domain protein). association endosomes mediated by its because deletion mutants...

To investigate the expression of SPARC and matrix metalloproteinases (MMPs) in normal conjunctiva pterygium tissues.This study involved paired control or uninvolved tissue from 21 patients. Quantitative real-time PCR was performed to assess MMP mRNA expression, whereas Western blot analysis protein levels tissue. Tissue localization SPARC, extracellular proteins, MMPs were determined by immunofluorescence analyses.SPARC transcript upregulated compared with conjunctiva. Immunofluorescence...

Purpose.: Pterygium is a prevalent ocular surface disease with unknown pathogenesis. The authors investigated the role of nuclear factor kappa B (NF-κB) transcription factors in pterygium. Methods.: Surgically excised primary pterygia were studied compared uninvolved conjunctiva tissues. NF-κB activation was evaluated using Western blot analysis, ELISA, and DNA-binding assays. Primary pterygium fibroblasts treated TNF-α (20 ng/mL), immunocytochemistry, phospho-IκBα stimulation target genes...

Failure of glaucoma filtration surgery (GFS) is commonly attributed to scarring at the surgical site. The human Tenon's fibroblasts (HTFs) are considered major cell type contributing fibrotic response. We previously showed that SPARC (secreted protein, acidic, rich in cysteine) knockout mice had improved success a murine model GFS. To understand mechanisms deficiency delaying subconjunctival fibrosis, we used gene silencing approach reduce expression HTFs and examined parameters important...

Purpose.: To investigate the effects of SLC4A11 gene depletion in human corneal endothelial cells. Methods.: achieve stable downregulation expression immortalized cells (HCECs), short-hairpin RNA (shRNA) targeted against was used. Cell growth and viability were determined using real-time cell analyzer trypan blue staining respectively. Apoptosis investigated by Annexin V TUNEL assays. Alterations apoptotic following silencing RT2Profiler PCR array for apoptosis while activation pathway...

Failure after glaucoma filtration surgery is attributed to fibrosis at the operated site. To understand wound healing process surgery, we have developed a mouse model for which closely mimics clinical response. In this study, describe systematic analysis of response in vivo. Our data revealed that post-surgical tissue was separable into two distinguishable phases. The early “acute inflammatory” phase characterized by significantly increased transcript expression Vegfa, Cxcl1, Cxcl5, Ccl2,...

Glaucoma Is a progressive optic neuropathy, which, If left untreated, leads to blindness. The most common and modifiable risk factor in glaucoma is elevated intraocular pressure (IOP), which can be managed surgically by filtration surgery. postoperative subconjunctival scarring response, however, remains the major obstacle achieving long-term surgical success. Antiproliferatives such as mitomycin C are commonly used prevent scarring. Efficacy of these agents has been tested extensively on...

Overproduction of type I collagen is associated with a wide range fibrotic diseases as well surgical failure such in glaucoma filtration surgery (GFS). Its modulation therefore clinical importance. Valproic acid (VPA) known to reduce variety tissues unclear mechanism action. In this report, we demonstrate that VPA inhibited production both conjunctival fibroblasts and the mouse model GFS. fibroblasts, decreased expression which intensified longer drug exposure suppressed steady-state...

Excessive accumulation of collagen is often used to assess the development fibrosis. This study aims identify genes that define fibrosis in conjunctiva following glaucoma filtration surgery (GFS). Using mouse model GFS, we have identified transcripts were upregulated fibrotic phase wound healing via RNA-seq. The top three highest induced belong Col8a1, Col11a1 and Col8a2. Further validation Col8a2 revealed their increase by 67-, 54- 18-folds respectively phase, compared 12-fold for Col1a1,...

Valproic acid (VPA) is a histone deacetylase inhibitor used clinically for neurological disorders. It also potentially useful as anti-fibrotic therapy it reduced collagen deposition in the post-operative conjunctiva. In this study, we further evaluated effects of VPA on inflammation using mouse model conjunctival scarring. VPA, injected into subconjunctiva immediately after surgery, did not cause any adverse tissue response when examined by live microscopy and produced an apparent reduction...

Abstract Background This study aimed to evaluate differences in iris gene expression profiles between primary angle closure glaucoma (PACG) and open (POAG) their interaction with biometric characteristics. Design Prospective study. Participants Thirty‐five subjects PACG thirty‐three POAG who required trabeculectomy were enrolled at the Singapore National Eye Centre, Singapore. Methods Iris specimens, obtained by iridectomy, analysed real‐time polymerase chain reaction for of type I collagen,...

Endofin is an endosomal protein implicated in regulating membrane trafficking. It characterized by the presence of a phosphatidylinositol 3-phosphate-binding FYVE domain positioned middle molecule. To determine its potential effectors or binding partners, we used carboxyl-terminal half endofin as bait to screen human brain cDNA library yeast two-hybrid system. Three clones that encode TOM1 were recovered. closely related VHS (VPS-27, Hrs, and STAM) domain-containing GGA family. Although...

Abstract Valproic acid (VPA) is a histone deacetylase inhibitor previously shown to promote the proliferation and self‐renewal of CD34 + hematopoietic cells. We tested effect VPA in conjunction with selective amplification technology developed by Viacell Inc. Stem cells enriched from frozen cord blood were cultured for 7 d, subjected reselection grown fresh medium further d. Treatment resulted an average two‐fold higher expansion CD45 34 compared control. Furthermore, VPA‐treatment induced...

TOM1 and its related proteins, TOM1-like1 (TOM1-L1) TOM1-like2 (TOM1-L2), constitute a subfamily of the VHS domain protein family. We have recently shown that endofin, FYVE associated with early endosome, is able to recruit cytosolic onto endosomal membranes. To reveal biological consequence endofin-mediated recruitment TOM1, we identified clathrin heavy chain as major interacting for TOM1. Optimal binding by involves three sites: residues 300-321, 321-326 putative clathrin-binding box at...

Purpose: Secreted protein acidic and rich in cysteine (SPARC) Hevin are structurally related matricellular proteins involved extracellular matrix assembly. In this study, we compared the anterior chamber biometric parameters iris collagen properties SPARC-, Hevin- SPARC-/Hevin-null with wild-type (WT) mice. Methods: The right eyes of 53 WT, 35 56 Hevin-, 63 mice were imaged using RTVue-100 Fourier-domain optical coherence tomography system. measured depth (ACD), trabecular-iris space area...