Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions

Experimental models for study of retinal pigment epithelial physiology and pathophysiology

Abstract We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions

Resonant Transfer and Excitation in Li-Like F Colliding with H₂

We have measured coincidences between x rays and projectiles that have captured one electron in F6+ + H2 collisions at projectile energies between 15 and 33 MeV. The cross sections for capture and simultaneous x-ray emission as a function of projectile energy show clear structures. Indications of an unexpectedly high population of high-n states predominantly formed by resonant transfer and excitation (RTE) were found. Above the Kln (n\u3e1) RTE resonance energies another maximum was observed

SDSS-RASS: Next Generation of Cluster-Finding Algorithms

We outline here the next generation of cluster-finding algorithms. We show how advances in Computer Science and Statistics have helped develop robust, fast algorithms for finding clusters of galaxies in large multi-dimensional astronomical databases like the Sloan Digital Sky Survey (SDSS). Specifically, this paper presents four new advances: (1) A new semi-parametric algorithm - nicknamed ``C4'' - for jointly finding clusters of galaxies in the SDSS and ROSAT All-Sky Survey databases; (2) The introduction of the False Discovery Rate into Astronomy; (3) The role of kernel shape in optimizing cluster detection; (4) A new determination of the X-ray Cluster Luminosity Function which has bearing on the existence of a ``deficit'' of high redshift, high luminosity clusters. This research is part of our ``Computational AstroStatistics'' collaboration (see Nichol et al. 2000) and the algorithms and techniques discussed herein will form part of the ``Virtual Observatory'' analysis toolkit.Comment: To appear in Proceedings of MPA/MPE/ESO Conference "Mining the Sky", July 31 - August 4, 2000, Garching, German

(R)-␣-Lipoic Acid Protects Retinal Pigment Epithelial Cells from Oxidative Damage

PURPOSE. To determine whether (R)-␣-lipoic acid (LA) protects cultured human fetal retinal pigment epithelial (hfRPE) cells against oxidative injury and identify the pathways that may mediate protection. METHODS. Cultured hfRPE cells were pretreated with various concentrations of LA for 14 to 16 hours followed by treatment with a chemical oxidant, tert-butylhydroperoxide (t-BuOOH; 0.8 mM, 3 hours). Reactive oxygen species (ROS) production and cell viability were measured using H 2 DCF and MTT assays, respectively. RPE cells were evaluated with fluorescent dyes (SYTOX Orange and SYTO Green; Molecular Probes, Eugene, OR), which differentiate between live and dead cells. Apoptosis was visualized by using the TUNEL assay. Changes in mitochondrial membrane potential were detected by JC-1 dye. Intracellular levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were measured by HPLC. Regulation of ␥-glutamylcysteine ligase (GCL), the rate-controlling enzyme of GSH production, was assayed by RT-PCR. RESULTS. Pretreatment of hfRPE cells with LA, 0.2 mM and 0.5 mM, significantly reduced the levels of t-BuOOH-induced intracellular ROS, by 23% and 49%, respectively. LA (0.5 mM) prevented oxidant-induced cell death and apoptosis and also increased the viability of oxidant-treated hfRPE cells from 38% to 90% of control. LA upregulated the mRNA expression of GCL, and was protective against t-BuOOH-induced decreases in both mitochondrial membrane potential and intracellular levels of GSH and GSH/GSSG. CONCLUSIONS. The present study suggests that the protective effect of LA involves multiple pathways and that LA could be effective against age-associated increase in oxidative stress and mitochondrial dysfunction in RPE cells. (Invest Ophthalmol Vis Sci. 2005;46:4302-4310) DOI:10.1167/iovs.04-1098 A ge-related macular degeneration (AMD) is the major cause of severe visual impairment for elderly individuals in developed countries. 1-3 Although the underlying pathogenic mechanisms and risk factors are not well understood, 1-9 epidemiologic studies suggest that environmental factors associated with oxidative stress-for example, cigarette smoking, sunlight exposure, and low dietary intake of antioxidants-are contributing factors in the development of AMD. 4 -7 Recent results have shown that antioxidants and zinc protect against the progression of the disease. 8 There is good evidence to suggest that the retinal pigment epithelium (RPE), choriocapillaris, and distal retina are all involved in this process, but the pathology associated with each can be difficult to distinguish, given their close anatomic, physiologic, and metabolic relationships. 7,15 A significant oxidative load is associated with the RPE phagocytosis of photoreceptor outer segments, 16 Cellular membranes with high polyunsaturated fatty acids are particularly susceptible to radical-induced chain reactions of lipid peroxidation. 17 Age-associated decreases in antioxidant defense mechanisms throughout the body can further increase the levels of RPE exposure to oxidants. For example, the levels of plasma glutathione (GSH), one of the major water-soluble antioxidants, decrease with age. 19 -21 Oxidantinduced mitochondrial dysfunction and death of RPE cells may contribute to the onset of AMD. 27-29 ␣-Lipoic acid is a potent intracellular antioxidant that can induce all three cellular protective mechanisms. 36 t-BuOOH is a relatively stable alkyl peroxide that readily penetrates cell membranes From th

Projectile Energy Loss in Multiply Ionizing Ion-Atom Collisions

The projectile energy loss for 7.5--25-MeV C6+,5+ and F6+ ions was measured for single collisions with He, Ne, Ar, and Kr as a function of the recoil-ion charge state and the projectile scattering. This energy loss was measured for collisions in which the projectile captured an electron and for those involving just direct ionization. We investigated and found a large average energy transfer (100--250 eV/electron) to the continuum electrons. A strong increase of the scattering angle with recoil-ion charge state was observed for both capture and direct ionization. The results imply that, for smaller impact parameters, higher recoil-ion charge states are produced and that higher energy losses are obtained. We observed a weak target-Z dependence of the energy loss. The results are compared with n-body classical-trajectory Monte Carlo calculations by Olson, semiclassical-approximation calculations by Schuch et al. [Nucl. Instrum. Methods Phys. Res. Sect. B 42, 566 (1989)], and the energy-deposition model

Advanced lost foam from casting technology

Previous research made significant advances in understanding the Lost Foam Casting (LFC) Process and clearly identified areas where additional research was needed to improve the process and make it more functional in an industrial environment. The current project focused on five areas listed as follows: Task 1: Precision Pattern Production Task 2: Pattern Coating Consistency Task 3: Sand Fill and Compaction Effects Task 4: Pattern Gating Task 5: Mechanical Properties of Castings. This report summarizes the work done under the current contract in all five areas in the period of October 1, 1994 through December 31, 1995. Twenty-eight (28) companies jointly participate in the project. These companies represent a variety of disciplines, including pattern designers, pattern producers, coating manufacturers, plant design companies, compaction equipment manufacturers, casting producers, and casting buyers

A GMBCG Galaxy Cluster Catalog of 55,424 Rich Clusters from SDSS DR7

We present a large catalog of optically selected galaxy clusters from the application of a new Gaussian Mixture Brightest Cluster Galaxy (GMBCG) algorithm to SDSS Data Release 7 data. The algorithm detects clusters by identifying the red sequence plus Brightest Cluster Galaxy (BCG) feature, which is unique for galaxy clusters and does not exist among field galaxies. Red sequence clustering in color space is detected using an Error Corrected Gaussian Mixture Model. We run GMBCG on 8240 square degrees of photometric data from SDSS DR7 to assemble the largest ever optical galaxy cluster catalog, consisting of over 55,000 rich clusters across the redshift range from 0.1 < z < 0.55. We present Monte Carlo tests of completeness and purity and perform cross-matching with X-ray clusters and with the maxBCG sample at low redshift. These tests indicate high completeness and purity across the full redshift range for clusters with 15 or more members.Comment: Updated to match the published version. The catalog can be accessed from: http://home.fnal.gov/~jghao/gmbcg_sdss_catalog.htm

GREAT3 results I: systematic errors in shear estimation and the impact of real galaxy morphology

We present first results from the third GRavitational lEnsing Accuracy Testing (GREAT3) challenge, the third in a sequence of challenges for testing methods of inferring weak gravitational lensing shear distortions from simulated galaxy images. GREAT3 was divided into experiments to test three specific questions, and included simulated space- and ground-based data with constant or cosmologically-varying shear fields. The simplest (control) experiment included parametric galaxies with a realistic distribution of signal-to-noise, size, and ellipticity, and a complex point spread function (PSF). The other experiments tested the additional impact of realistic galaxy morphology, multiple exposure imaging, and the uncertainty about a spatially-varying PSF; the last two questions will be explored in Paper II. The 24 participating teams competed to estimate lensing shears to within systematic error tolerances for upcoming Stage-IV dark energy surveys, making 1525 submissions overall. GREAT3 saw considerable variety and innovation in the types of methods applied. Several teams now meet or exceed the targets in many of the tests conducted (to within the statistical errors). We conclude that the presence of realistic galaxy morphology in simulations changes shear calibration biases by $\sim 1$ per cent for a wide range of methods. Other effects such as truncation biases due to finite galaxy postage stamps, and the impact of galaxy type as measured by the S\'{e}rsic index, are quantified for the first time. Our results generalize previous studies regarding sensitivities to galaxy size and signal-to-noise, and to PSF properties such as seeing and defocus. Almost all methods' results support the simple model in which additive shear biases depend linearly on PSF ellipticity.Comment: 32 pages + 15 pages of technical appendices; 28 figures; submitted to MNRAS; latest version has minor updates in presentation of 4 figures, no changes in content or conclusion