Abstract:
PURPOSE:Interphotoreceptor retinoid-binding protein (IRBP) binds and transports retinoids and fatty acids in the interphotoreceptor space (IPS). To understand the relationship between the protein structure and its functions requires bulk quantities of human IRBP. The authors sought to produce recombinant human IRBP (rhIRBP), a perfect duplicate in amino acid sequence of the authentic human protein. This material could serve as a supply of the protein and later could be used to make mutants of the protein. The goals of the present study were to produce human IRBP in an expression system and to examine some of its biochemical properties. METHODS:A cDNA encoding human IRBP was cloned into the transplacement vector, pVL1392, and the plasmid was recombined with linearized baculovirus on cotransfection into Sf9 cells. Viruses containing the human IRBP cDNA were identified by polymerase chain reaction analysis. IRBP was secreted from virus-infected insect cells. rhIRBP was purified from cell medium and was examined by chromatography, N-terminal protein sequencing, immunologic techniques, and fluorometry. Eyecup and retina washes of human donor eyes provided a source of authentic human IRBP (IPS-IRBP). RESULTS:rhIRBP and IPS-IRBP exhibit similar elution profiles on concanavalin A, ion-exchange, and size exclusion chromatography. rhIRBP contains a five-amino-acid propeptide at the N-terminus as deduced from the cDNA sequence. Retinol binding of rhIRBP has been characterized by fluorometric titration. The dissociation constant is approximately 1.04 microM, close to that reported for bovine IRBP. By scanning fluorometry, the emission and excitation maxima are 479 nm and 339 nm, respectively. CONCLUSIONS:The baculovirus system provides an excellent method to produce and secrete human IRBP. The recombinant protein can be readily purified from cell culture medium. Its behavior in chromatography and in binding studies suggests that the recombinant protein is virtually identical to the authentic protein. This validates its use in place of IRBP from human donor eyes. Small, but significant, differences in biochemical properties in comparing human and bovine material highlight the significance of studying the human protein.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Lin ZY,Si JS,Nickerson JMsubject
Has Abstractpub_date
1994-09-01 00:00:00pages
3599-612issue
10eissn
0146-0404issn
1552-5783journal_volume
35pub_type
杂志文章abstract:PURPOSE:Anterograde intraflagellar transport (IFT) is essential for photoreceptor outer segment formation and maintenance, as well as for opsin trafficking. However, the role of retrograde IFT in vertebrate photoreceptors remains unclear. The purpose of this study was to evaluate zebrafish photoreceptors lacking the re...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.09-3828
更新日期:2009-11-01 00:00:00
abstract:PURPOSE:To evaluate the ability of various screening tests, both individually and in combination, to detect glaucoma in the general Latino population and high-risk subgroups. METHODS:The Los Angeles Latino Eye Study is a population-based study of eye disease in Latinos 40 years of age and older. Participants (n = 6082...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.09-5126
更新日期:2011-08-05 00:00:00
abstract:PURPOSE:To determine whether anthocyanin-enriched bilberry extracts modulate pre- or posttranslational levels of oxidative stress defense enzymes heme-oxygenase (HO)-1 and glutathione S-transferase-pi (GST-pi) in cultured human retinal pigment epithelial (RPE) cells. METHODS:Confluent ARPE-19 cells were preincubated w...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.06-0452
更新日期:2007-05-01 00:00:00
abstract:Purpose:Dry eye disease (DED) is a multifactorial disease associated with ocular surface inflammation. Toll-like receptors (TLRs) are integral in the initiation of inflammatory signaling. Therefore, we evaluated the effect of TLR-deficiency on dry eye-related ocular surface damage and inflammation using a mouse model o...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.17-23397
更新日期:2018-06-01 00:00:00
abstract:Purpose:Aging affects a variety of visual functions. In this study, we aim to quantitatively investigate the temporal characteristics of visual processing in aging. Methods:Twelve younger (24.1 ± 1.6 years) and 12 older observers (58.4 ± 3.6 years) participated in the study. All participants had normal or corrected-to...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.61.5.60
更新日期:2020-05-11 00:00:00
abstract:PURPOSE:To explore whether pointwise rates of visual field progression group together in patterns consistent with retinal nerve fiber layer (RNFL) bundles. METHODS:Three hundred eighty-nine eyes of 309 patients from the Advanced Glaucoma Intervention Study with ≥6 years of follow-up and ≥12 reliable visual field exams...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-9021
更新日期:2012-04-30 00:00:00
abstract:PURPOSE:To apply tissue-engineered cell sheet transplantation after excimer laser keratectomy as a novel approach for the reduction of postoperative corneal haze. METHODS:Limbal biopsy specimens were obtained, and epithelial cells were cultured on temperature-responsive culture inserts without the use of feeder cells....
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-0995
更新日期:2006-02-01 00:00:00
abstract::Experimental esotropia was produced surgically in infant rhesus monkeys (Macaca mulatta) for brief periods of time. Electrophysiological studies of the visual cortex showed that esotropia of only 2 weeks duration in an infant monkey is sufficient to cause a marked shift of dominance in favor of the fixating eye and to...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1979-05-01 00:00:00
abstract:PURPOSE:CD4(+)CD25(+)FoxP3(+) naturally occurring regulatory T cells (Tregs) maintain self-tolerance and function to suppress overly exuberant immune responses. However, it is unclear whether innate immune cells modulate Treg function. Here the authors examined the role of innate immunity in lymphomyeloid homeostasis. ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.10-6536
更新日期:2011-04-08 00:00:00
abstract:PURPOSE:To evaluate the dynamic changes in refraction during the accommodative facility test in myopes and emmetropes. METHODS:Ten myopes and 10 emmetropes participated in the study. All were young adults, and refractive error in the myopes was corrected with soft contact lenses. Monocular accommodative facility measu...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.07-0269
更新日期:2007-09-01 00:00:00
abstract::Photoreceptor-mediated mechanisms were studied in patients with a recently identified retinopathy typified by night blindness, cystoid maculopathy, and similar scotopic and photopic electroretinograms (ERGs). Dark-adapted spectral sensitivity functions were only partly explained as composites of rod and cone curves sh...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1990-05-01 00:00:00
abstract::An objective visual field can be mapped in terms of stimulus-induced eye movement. The authors used the scleral coil technique to record vergence and conjugate eye movements while stimulating different visual field locations with a 3 X 3 deg target whose image vergence was oscillated. For each of three subjects tested...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1986-05-01 00:00:00
abstract:PURPOSE:We investigated the response of retinal vessel diameters and oxygen saturation to flicker light stimulation of neuronal activity in patients with diabetic retinopathy. METHODS:We included 18 patients with nonproliferative diabetic retinopathy (mean age 62.2 ± 8.3 years, diabetes type 1 in 4 patients and type 2...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-9659
更新日期:2012-06-26 00:00:00
abstract::To investigate the role of cellular immunodeficiency in recurrent toxoplasmic retinochoroiditis, six Cynomolgus monkeys (Macaca fascicularis) with healed toxoplasmic lesions of the retina were immunosuppressed by total lymphoid irradiation. Three months prior to irradiation 30,000 Toxoplasma gondii organisms of the Be...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1988-06-01 00:00:00
abstract:PURPOSE:We examined the treatment period necessary to restore retinal and visual stability following trauma to the optic nerve. METHODS:Cats received unilateral optic nerve crush and no treatment (NT), treatment of the injured eye with brain-derived neurotrophic factor (BDNF), or treatment of the injured eye combined ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.13-12683
更新日期:2013-10-09 00:00:00
abstract::Patients with choroidal melanomas and simulating choroidal tumors were tested in a masked manner in a leukocyte-migration inhibition (LMI) assay with a 3M KCl antigen derived from a metastatic choroidal melanoma. Good discrimination petween patient groups was observed with this antigen. The sensitivity and specificity...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1977-02-01 00:00:00
abstract:PURPOSE:To evaluate the differential effects of ganglion cell depletion from experimental glaucoma on the relative metabolic activities of neurons in the parvo (P)- and magno (M)-cellular visual pathways of the macaque visual system. METHODS:Monocular experimental glaucoma was induced in monkeys (Macaca mulatta and M....
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:2000-06-01 00:00:00
abstract:PURPOSE:The effects of adrenergic agonists and antagonists on Na+, K+, Cl- cotransport in fetal human nonpigmented ciliary epithelial (NPE) cells were investigated. METHODS:86Rb+ as a marker for K+ was used to study ouabain-insensitive, bumetanide-sensitive 86Rb+ uptake in cultured NPE monolayers. Cyclic adenosine mon...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1996-05-01 00:00:00
abstract:Purpose:Lens epithelial cell (LEC) conversion to myofibroblast is responsible for fibrotic cataract surgery complications including posterior capsular opacification. While transforming growth factor beta (TGFβ) signaling is important, the mechanisms by which the TGFβ pathway is activated post cataract surgery (PCS) are...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.18-25067
更新日期:2018-10-01 00:00:00
abstract::Transmembrane electrical measurements were performed on the isolated rabbit iris-ciliary body (I-CB) to study the direct effects of halogenated inhalation anesthetic agents on the ciliary epithelium. Addition of either halothane, enflurane, or isoflurane to the control 95% O2:5% CO2 gas mixture resulted in a dose-depe...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1991-05-01 00:00:00
abstract:PURPOSE:To investigate the effect of K-115, a novel Rho kinase (ROCK) inhibitor, on retinal ganglion cell (RGC) survival in an optic nerve crush (NC) model. Additionally, to determine the details of the mechanism of K-115's neuroprotective effect in vivo and in vitro. METHODS:ROCK inhibitors, including K-115 and fasud...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.13-13842
更新日期:2014-10-02 00:00:00
abstract:PURPOSE:Aquaporin (AQP) is a hexahelical integral membrane protein that functions as a constitutive channel for water and regulated channel for cations in fluid transporting tissues, including many in the eye. Although AQP1 has been cloned from a cDNA library prepared from cultures of retinal pigment epithelial (RPE) c...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.03-0001
更新日期:2003-06-01 00:00:00
abstract:PURPOSE:The purpose of this study was to examine the relationship between objectively measured ambient light exposure and longitudinal changes in axial eye growth in childhood. METHODS:A total of 101 children (41 myopes and 60 nonmyopes), 10 to 15 years of age participated in this prospective longitudinal observationa...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-15978
更新日期:2015-10-01 00:00:00
abstract::Analysis of cell populations in the cornea may be performed rapidly and accurately employing the technique of enzymatic disaggregation. To illustrate this method normal rat corneas and corneas infected 24 and 48 hours previously with Staphylococcus aureus were disaggregated in a solution containing pancreatin and coll...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1983-02-01 00:00:00
abstract:PURPOSE:To investigate the induction of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) gene expression by prostaglandin E2 (PGE2) in cultured rat Müller cells and to study the mechanism of the induction. METHODS:Müller cells were obtained from neonatal Sprague-Dawley rat retinas an...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1998-03-01 00:00:00
abstract:PURPOSE:To investigate the application of anterior corneal and ocular aberrations in detecting mildly ectatic corneas. METHODS:This study retrospectively reviewed the data of 220 eyes separated into three groups by the NIDEK Corneal Navigator System automated corneal classification software: normal (N) (n = 123); form...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-8803
更新日期:2012-05-17 00:00:00
abstract:PURPOSE:To identify the risk factors for having advanced glaucomatous visual field loss on the first visit at three hospital eye services. METHODS:This was a hospital-based, case-control study involving patients newly diagnosed with glaucoma at first visit to one of three ophthalmic departments in the United Kingdom. ...
journal_title:Investigative ophthalmology & visual science
pub_type: 临床试验,杂志文章,多中心研究
doi:
更新日期:1999-09-01 00:00:00
abstract:PURPOSE:A human biovar of Chlamydia trachomatis was used to develop a murine model of ocular chlamydial infection. The inbred mouse model will allow detailed immunologic studies during ocular infection, and use of a human biovar for infection may aid in identification of appropriate vaccine strategies against chlamydia...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1995-09-01 00:00:00
abstract:PURPOSE:This study was performed to investigate motion perception in patients with glaucoma. METHODS:A random dot motion test was used to measure three aspects of central motion perception: minimum displacement threshold (Dmin), maximum displacement threshold (Dmax), and coherence threshold (signal to noise). Motion p...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1993-12-01 00:00:00
abstract:PURPOSE:To study the putative role of endogenous matrix metalloproteinases (MMPs) in retinal neovascularization, an established mouse model was used to compare the retinal neovascularization observed in wild-type mice with that in mice without the MMP-2 or -9 genes. METHODS:C57Bl/6 (MMP-2(+/+) and -9(+/+)), MMP-2-defi...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.03-0249
更新日期:2003-12-01 00:00:00