National Eye Institute U * I *««* . _ , LIBRARY SEP 1 1991 National Institutes of Health Annual Report of Intramural Research October 1, 1989 to September 30, 1990 / J DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00248-03 LI PERIOD COVERED October 1, 1989 to September 30, 1990 irTLE OF PROJECT {SO charaaers or less. 77tfe must Ht on one line between the borders.) Magainin Therapy of Infectious Keratitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Rubens Belfort, Jr. M.D.,Ph.D. Visiting Scientist LI.NEI Others: Robert B. Nussenblatt M.D. Clinical Director LI, NEI Rashid Mahdi Biologist LI, NEI Juan Lopez M.D. Visiting Associate LI, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical ImTnunoregulation INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 8.0 PROFESSIONAL: 6.0 OTHER: 2.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (b) Human tissues KI (c) Neither n (a1) Minors D (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Studies are being conducted in animals to determine the in vivo activity of a new class of antimicrobial peptides isolated from the skin of the African frog Xenopus laevis and called magainins. The family of peptides consists of two closely related peptides, each 23 amino acids, that inhibit growth of numerous species of bacteria and fungi in vitro. An anim al model of experimental bacterial keratitis induced in adiilt New Zealand white rabbits were used to determine the in vivo relevance of the antimicrobial activity of magainins. Pseudomonas aeruginosa was primarily considered because it is the most destructive and the most difficult to treat corneal infection in humans. Topical treatment with magainin drops was started immedi- ately after the infection. The control animals were treated with the vehicle or with gentamycin. These studies have shown potential activity of the magainin in the treatment oi Pseudomonas corneal ulcers. Although the animals coidd tolerate the treatment well, magainin drops induced severe chemosis with conjunctival hyperhemia which can aggravate the inflammation related to the infection. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED PROJECT NUMBER ZOl EY 00265-01 LI October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the tmrders.) T Cell Vaccination in Experimental Autoimmune Uveitis PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Evelyne Beraud Ph.D. Visiting Associate LI, NEI Others: Satoshi Kotake M.D. Chi-Chao Chan Ph.D. Stephen M. Oddo M.D. Rachel R. Caspi Ph.D. Visiting FeUow Medicm Officer Special Volvinteer Visiting Scientist LI, NEI LI, NEI LI, NEI LI, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI, NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 1.41 PROFESSIONAL: 1.41 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues IXl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Autoimmune T Ijrmphocytes can be used under appropriate conditions to induce resistance to the specific autoimmune disease that they usually produce. This practice, termed "T cell vaccination," has been successful in other experimental autoimmune diseases. Our investiga- tions revealed that experimental autoimmune uveitis in rats can be down-regulated by this procedure. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00245-03 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 characters or less. Title must lit on one line between the borders.) Molecular Biology of Cataracts PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Teresa Borras Ph.D. Biologist LMOD.NEI Others: Ignacio Rodriguez Ph.D. Pedro Gonzalez Ph.D. Carlos Hemandez-CalzadiUa Ph.D. Staff Fellow Visiting Fellow Special Volunteer LMOD, NEI LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) Department of Chemistry, KaroHnska Institute, Stockhohn, Sweden (Dr. Hans Jomvall, Ph.D.); Diabetes Programs Branch, National Institute of Diabetes and Digestive and Kidney Diseases, NIH (Flora de Pablo, M.D.) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Cataracts INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 3.0 PROFESSIONAL: 2.8 OTHER: 0.2 CHECK APPROPRIATE BOX(ES) D (a) Human subjects □ (a1) Minors D (a2) Interviews n (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Investigation of the molecular mechanisms of hereditary cataracts using as a model the nuclear hereditary cataract of strain 13/N guinea pigs continues. This year, the focus was on the further characterization of the ^-crystallin-cDNA present in the lens of the cataractous animal. We developed a new method to construct cDNA libraries from small amoimts of tissue and obtained a cDNA library from four heterozygoxis guinea pig lenses. Screening of this library produced normal and defective ^-cDNAs. Sequencing of the cataractous ^-cDNA revealed a 102 bp deletion corresponding exactly to 34 amino adds. The deletion, which does not interfere with the reading frame, is located toward the carboxy end of the protein. Analysis of the genomic DNA from normal and affected animals indicated that the deleted mRNA region is present in the genome of the cataractous animals. This result suggests that a mutation affecting the sphcing mechanism is possibly the cause of the mRNA deletion. In studies of the role of the zeta mRNA in the Uver of guinea pigs, we isolated four clones from an adult Uver cDNA Hbrary. Upon sequencing, two of the clones showed the poly A tail at different positions in the mRNA, confirming different processing of the ^-crystallin gene in the liver versus the lens. Sequencing the coding region revealed that the amino acids of the liver protein are identical to those of the lens protein. Having successfully cloned the complete C-crystallin gene, we are now analyzing it. Thus far, we have characterized six introns and a very long exon comprising the untranslated 3' end of the mRNA. Sequencing one of the clones has revealed the presence of one exon that corresponds exactly to the deleted region of the zeta mRNA of the cataractous animals. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00244-03 LSR TITLE OF PROJECT {80 characters or less. We must lit on one line between the borders.) Ociilomotor and Visual Disorders in Humans PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute atliliation.) PI: James R. Carl M.D. Expert LSR, NEI Others: Edmond J. FitzGibbon Michael E. Goldberg Michael J. Todd M.D. Senior Staff FeUow LSR, NEI M.D. Chief NMS LSR, NEI Special Volunteer LSR, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Neuro-Ophthalmologic Mechanisms Section INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 1.4 PROFESSIONAL: 1.4 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) S (a) Human subjects n (a1) Minors D (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Patients with a variety of oculomotor and visual problems were evaluated by clinical examination and high resolution eye movement recordings. Patients with cortical and cerebellar disease were evaluated for amount and type of clinical abnormality. Their eye movements in response to visual stimuli were tested using paradigms devised to test the various oculomotor sub-systems, which were correlated with the clinical findings. Patients with cerebellar deficits showed the expected saccadic dysmetria to visual targets and low gain pxirsuit. In addition, preliminary data suggest that some cerebellar patients cannot change the gain of their saccades. Visual motion processing by normal subjects was evaluated using paradigms that tested the subjects' ability to predict futxire target motion in the generation of saccadic and smooth piirsuit eye movements. Smooth pursuit in hvunans was foiind to be primarily based on prediction of future target motion and not on negative feedback to reduce retinal sUp. A nuanber of patients with unilateral cerebral hemispheric lesions showed pursuit deficits but had normal saccades to moving targets, suggesting that their lesions were not those of visual motion processing but actually of motor control. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00201-06 LMOD PERIOD COVERED October 1. 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must tit on one line between the borders.) Structure, Expression and Gene Complexity of Aldose Reductase PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Deborah Carper Ph.D. Biologist LMOD,NEI Others: Susan Old Ph.D. Staff FeUow CaroHne Graham B.A. Chemist LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) National Children's Medical Research Center, Tokyo, Japan (Chihiro Nishimiira, M.D.) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Cataracts INSTITUTE AND LOCATION NEI. NIH. Bethesda. MD 20892 TOTAL MAN-YEARS: 3.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors D (a2) Interviews PROFESSIONAL: 2.0 OTHER: 1.0 n (b) Human tissues K](c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Aldose reductase (AR) is believed to play a major role in the secondary complications of diabetes. Studies in this laboratory have focused on the regulatory properties, potential inhibitor sites, and overall structure of aldose reductase. Better knowledge of these aspects of the protein should help in regulating the xindesirable effects of this enzyme. For example, expression of large amoiints of recombinant AR protein now set the stage for x-ray crystallography and site-directed mutagenesis studies with the goal of locaUzing the important functional sites of the molecvde. Regulation of the gene for AR is also being studied. Controlling the expression of AR at the gene level may limit the effect this protein has on the accumulation of sorbitol in hyperglycemic conditions. Rat lens and hximan placenta AR have been expressed in Escherichia coli. The recombinant proteins have been purified and have immionological and kinetic properties similar to their respective tissue AR, including the same substrate and inhibitor profiles. Characterization of the promoter area of the AR gene shows one TATA and two CCAAT boxes and an approximately 14 kb gene. The nucleotide sequence is nearly complete. We have found that AR is induced under hypertonic conditions in several ceU types, including cultured dog lens epitheUal cells and dog kidney endothelial cells. Recently, we have also found an 8- to 16-fold induction of oB-crystalhn in these same cells grown in hypertonic media (550 mosM). It appears that AR and oB-crystallin are osmotic stress proteins. PHS 6040 (Rev. 1/84) GPO 914-916 DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00123-10 OGCS PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Tide must fit on one line between &ie borders.) Clinical Psychophysics of the Visual System PR INCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Rafael Caruso M.D. Visiting Scientist OGCS,NEI Others: Muriel I. Kaiser-Kupfer M.D. Chief Doris J. Collie A.A. Clinical Research TechTiinan Maria Bankiewicz M.D. Visitiag FeUow ClinicaJ Research Patricia A. Mercer M.P.A. Technician OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) Georgetown University Center for Sight, Washington, DC (Despina Koutsandreas, B.S., Le- anne Renter, B.S.) and School of Optometry, University of California at Berkeley (Jay M. Enoch, Ph.D., Richard Knowles). LAB/BRANCH Ophthalmic Genetics and Chnical Services Branch SECTION Clinical Services Section INSTITUTE AND LOCATION NEI, Nm, Bethesda, MP 20892 TOTAL MAN- YEARS: 0.55 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 0.55 OTHER: 0.0 n (b) Human tissues IS (a) Human subjects D (a1) Minors D (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) n(c) Neither The visual function of patients with ocular diseases or lesions iu the visual pathways and of normal subjects is measured using psychophysical techniques. The data obtained are correlated with those obtained by electrophysiological testing of visual function. The results, which contrib- ute to the diagnosis of ocular and neural disorders that affect vision, are needed to characterize the nature and evolution of the disorders. They are also valuable ia the assessment of the effect of treatment regimens on the outcome of these diseases. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00144-09 OGCS PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between the borders.) Clinical Electrophysiology of the Visual System PRINCIPAL INVESTIGATOR (List other professional personnel below ttie Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Rafael Caruso M.D. Visiting Scientist OGCS.NEI Others: Muriel I. Kaiser-Kupfer M.D. Maria A. Bankiewicz M.D. Patricia A. Mercer M.P.A. Doris J. Collie A.A. Chief Visiting Fellow Clinical Research Tech. Clinical Research Tech. OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) Center for Sight, Georgetown University, Washington, DC (Despina Koustsandreas, B.S., Le- anne M. Renter, B.S.) LAB/BRANCH Ophthalmic Genetics and Chnical Services Branch SECTION Clinical Services Section INSTITUTE AND LOCATION NEI. Nm, Bethesda. MD 20892 TOTAL MAN- YEARS: 1.10 PROFESSIONAL: 0.55 OTHER: 0.55 CHECK APPROPRIATE BOX(ES) IXI (a) Human subjects H (al) Minors n {a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The visual function of patients with ocular diseases or lesions in the visual pathways and of normal subjects is measiired objectively with electrophysiological techniques. These data are correlated with those obtained with psychophysical tests of visual function. The results obtained contribute to the diagnosis of ocular and neural disorders that affect vision, and are needed to characterize their nature and evolution. They are also valuable in the assessment of the effects of different forms of treatment on the outcome of these diseases. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00257-02 OGCS PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Tide must fit on one line between the tmrders.) Visual Function Diagnosis Service PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Inveso'galor.) (Name, title, laboratory, and Institute affiliation.) PI: Rafael Caruso M.D. Visiting Scientist OGCS,NEI Others: Muriel I. Kaiser-Kupfer M.D. Chief OGCS, NEI COOPERATING UNITS (if any) Center for Sight, Georgetown University, Washington, DC (Donna C. Optican, M.A.S., Despina Koutsandreas, B.S., Leanne M. Renter, B.S.) LAB/BRANCH Ophthalmic Genetics and Chnical Services Branch SECTION Eye Services Section INSTITUTE AND LOCATION NEI, NIH, Bethesda. MP 20892 TOTAL MAN-YEARS: 0.1 PROFESSIONAL: 0.1 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) E (a) Human subjects n (a1) Minors D (a2) Interviews n (b) Human tissues n(c) Neither SUMMA RY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This general service project provides diagnostic support for all research protocols conducted by the cUnical sections of the National Eye Institute and other Institutes requiring assessment of visual function. Psychophysical and electrophysiological techniques are used to detect and quantify vision loss due to disorders of the ocular media, uvea, retina, optic nerve, and central visual pathways. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00184-08 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between tt)e borders.) Cellular Mechanisms in Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute attiliation.) LI,NEI LI,NEI LI,NEI LI,NEI LI,NEI LI,NEI LI,NEI LI,NEI PI: Rachel R. Caspi Ph.D. Visiting Associate Others: Stephen Oddo M.D. Special Volunteer Horst Helbig M.D. Special Volunteer Sina Bahmanyar M,D. Staff FeUow Chi-Chao Chan M.D. Medical Officer William T^eake M.S. Biologist Barry Gnibbs B.S. Biologist Yujiro Fujiao M.D. Visiting Fellow COOPERATING UNITS (ilany) Immunology Research Unit, Klinikum Stegliz, Frele Universitat Berlin, Federal Republic of Germany (Tibor Diamantstein, Ph.D., Head); Arthritis and Rheimiatism Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases (Ronald L. Wilder, Head) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 1.45 CHECK APPROPRIATE BOX(ES) □ (a) Human subjects n (a1) Minors D (a2) Interviews PROFESSIONAL: 1.43 OTHER: 0.02 n (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Cellular mechanisms of ociilar immiinologically mediated disease are being studied in animal models of experimental autoimmune uveoretinitis. In vivo functional long-term T cell lines and T cell clones are developed and maintained in vitro from lymphoid organs of experimental animals immunized with uveitogenic ocular proteins. The phenotype and functional properties of these cells, as well as their interaction with ocular resident cells are being studied. The goal of these studies is to identify the immunoreactive cells and mediators as well as the pathogenic mechanisms involved in the intraocular inflammatory process. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00258-02 U TfTLEOF PROJECT {80 characters or less. We must lit on arte line betweer) the borders.) Experimental Autoimmiine Uveitis in the Mouse PRINCIPAL INVESTIGATOR (List other prolessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute attiliation.) PI: Rachel R. Caspi Ph.D. Visiting Associate LI.NEI Others: Barry Grubbs Chi-Chao Chan B.S. Biologist M.D. Medical Officer WilliaTn T,eake M.S. Biologist Benjamin Rubin M.D. Staff Fellow LI,NEI LI,NEI LI,NEI LI,NEI COOPERATING UNITS (ilany) Laboratory of Immunobiology, Rega Instituut, Katholieke Universiteit Leuwen, Belgiimi (A. Billiau, M.D., Head) LAB/BRANCH Laboratory of Immunology SECTION Section on Immimoregulation INSTITUTE AND LOCATION NEL Nm, Bethesda, MP 20892 TOTAL MAN- YEARS: 0.84 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors n (a2) Interviews PROFESSIONAL: 0.52 OTHER: 0.32 D (b) Human tissues ISl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) A new model of experimental autoimmune uveitis (EAU) is being developed in the mouse species, which has until now been considered refractory to induction of ocular autoimmimity. Different retinal antigens, as well as various immunization protocols, are being evaluated for efficacy of EAU induction. The pathological manifestations, disease coiirse, and genetic back- groimd of susceptibility to disease in murine EAU are being studied in relationship to the induction protocol. The goal of these studies is to estabUsh in the mouse species a rodent model of EAU that offers some important advantages over other rodent models of EAU. The extensive knowledge of the immunological parameters of the mouse and the availabihty of genetically defined strains wiU be of great value in the study of celltdar mechanisms and immimogenetics of ocular autoimmune disease. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00124-10 LRCMB TITLE OF PROJECT (flO characters or less. Title must fit on one line between the borders.) Metabolism of the Retina and Pigment Epithelium PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Gerald J. Chader Ph.D. Chief LRCMB, NEI Others: Robert Waldbillig R. Theodore Fletcher Ph.D. M.S. Expert Chemist LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) LAB« RANCH Laboratory of Retinal Cell and Moleciilar Biology SECTION Section on Gene Regulation INSTITUTE AND LOCATION NEI, NIH, Bethesda. MP 20892 TOTAL MAN-YEARS: 1.7 CHECKAPPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors n (a2) Interviews PROFESSIONAL: 1.2 OTHER: 0.5 12 (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) InsuJin and insiihn-like growth factor- 1 (IGF-1) are abundant in ocular tissues. They may act as messengers, coding for differentiation in the eye, may be directly or indirectly involyed in the visual process in the adult animal or control other important processes such as outer segment renewal. Equally important, receptors for these messengers are highly concentrated in devel- oping retina, pigment epithehum, and sclera, and may play a role in differentiation of these tissues. All the components of an IGF-1 system are present in the retinal interphotoreceptor matrix (IPM). The photoreceptor region thus possesses an IGF-1 autocrine-paracrine system. Abnormal ocular growth during early retinal degeneration and experimental myopia, for exam- ple, could be associated with changes in insulin and IGF-1 concentrations, or possibly with binding to receptors and/or binding proteins. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00148-17 LRCMB TITLE OF PROJECT {80 charaaers or lass. Title must fit on one iine between the borders.) Visual Control Mechanisms PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Gerald J. Chader Ph.D. Chief LRCMB, NEI Others: R. Theodore Fletcher Lila Inouye Betty J. Hayden M.S. M.D. Ph.D. Chemist Staff FeUow Staff FeUow LRCMB, NEI LRCMB, NEI LRCMB, NEI COOPERATING UNITS (ilany) School of Veterinary Medicine, University of Pennsylvania (G. Aguirre, D.V.M., Ph.D.); De- Bartment of Anatomy, Erasmus University, Rotterdam, The Netherlands (S. Sanyal, Ph.D.); >epartment of Zoology, University of Lund, Lund, Sweden (T. van Vee n, Ph.D.) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on G^ne Regulation INSTITUTE AND LOCATION NEL NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 2.7 PROFESSIONAL: 2.2 OTHER: 0.5 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews 12 (b) Human tissues n(c) Neither SUMMARY OF W/ORK {Use standard unreduced type. Do not exceed the space provided.) Retinitis pigmentosa and retinoblastoma are important hereditary diseases that usually cause retinal dysfunction in the first decade or two of life. Thus, it is important to study genes and their protein products specific to the retina that may be abnormal either in function or concentration in these retinal diseases. As a model system for photoreceptor-specific proteins, we are studying the regulation of expression of interphotoreceptor retinoid-binding protein (IRBP), a retinoid-transport protein synthesized by the photoreceptor neuron and secreted into the interphotoreceptor matrix. In a second study, we have found that a specific cAMP-dependent protein kinase exhibits a defect in synthesis in human retinoblastoma t\imor cells that is similar in osteosarcoma. Such a defect could cause the uncontrolled growth of retinoblastoma ceUs. In a third study, we have excellent new evidence for the general hypothesis of "transdifferentiation" because we have found that retinoblastoma cells highly express specific lens crystallins in vivo and in vitro. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00222-05 LI PERIOD COVERED October 1. 1989 to September 30, 1990 TITLE OF PROJECT (80 characters or less. Title must fit on one line between the txirders.) Immunopathology in the Eyes with Experimental Autoimimine Uveitis (EAU) Kir^inAI IMl/CCTiriATr\D // ;^» a^k^^ n^A/»»»;..»/ - ; i._(_,. ti — n-- .__( / ^- ._-./« I ^■^_ (_i , ■ ■ — PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Chi-ChaoChan M.D. Medical Officer LI.NEI Others: Robert B. Nussenblatt IgalGery Rachel R. Caspi MingNi M.D. Clinical Director NEI Ph.D. Head, Section on LI, NEI Experimental Immunology Ph.D. Visitiag Associate LI, NEI M.D. Visiting Fellow LI, NEI COOPERATING UNITS (if any) University of Tokyo, School of Medicine (Manabu Mochizuld, M.D.); University of Montreal, School of Medicine (Francois G. Roberge, M.D.) LAB^RANCH Laboratory of Immunology SECTION Section on Immunoresrulation INSTITUTE AND LOCATION NEI. NIH, Bethesda. MP 20892 TOTAL MAN-YEARS: 1.15 PROFESSIONAL: 1.15 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues ia(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Identity and topographic localization of immimocompetent cells and alteration of surface markers on ocular resident cells in rodents with experimental autoimmune uveoretinitis (EAU) were analyzed by immunohistochemical studies. In the early stage of EAU, T helper/inducers were the predominant cells in the eye; in the late stage, a relative increase of T suppressor/cjrtotoxic cells was observed. T lymphocjrte specificity is directed to small fragments of antigen boimd to ceU surface major histocompabihty complex (MHC) molecules which are presented on the surface of specialized antigen-presenting cells. Expression of MHC class 11 antigens was observed on ocular resident cells such as RPE, retinal endothelixmi, keratocytes, fibroblasts, and ciliary epitheUum. Both the infiltrate ing cell subpopulation and the expression of class 11 antigens on ocular resident cells can be altered by various immunomodulating agents, some of which — Qingaining, FK506, 15-deoxysperguaIine, interleukin 2-PE40, and S-antigen — ^were evaluated for their immunopathological effects on EAU. Clinical and immunopathologic examinations of the eyes of mice with EAU demostrated a chronic relapsing focal granulomatous inflammation and vascuHtis in the retina and choroid. Development of subretinal neovascularization may also occur. The infiltrating cells were mainly macrophages and T helpers/inducers (CD4). The expression of MHC class II antigens is confined to ocvdar resident cells immediately at the inflammatory sites. Due to the chronicity and recurrence of EAU, the mouse may serve as a better model for himoan ocular inflammation. Experimental endotoxin-induced uveitis (EIU) is a model for anterior uveitis in humans. The mechanism of this inflammation is still unclear. Because recentiy it has been suggested that activation of phospholipase A2 may play a role in the initiation and propagation of this disease, we studied the effect of a novel synthetic peptide £ind a PLA2 inhibitor, antiflammin, on EIU in rats. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOIEY 00224-04 LI TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Sympathetic Ophthalmia: Immunopathological Findings PRINCIPAL INVESTIGATOR (Ust other protessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Chi-ChaoChan M.D. Medical Officer LI,NEI Others: Robert B. Nussenblatt Alan G. Palestine Toichiro Kuwabara M.D. Clinical Director NEI M.D. Head, Section on LI, NEI Clinical Immunology M.D. Chief, Laboratory of LI, NEI Ophthalmic Pathology COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI, Nm, Bethesda. MP 20892 TOTAL MAN- YEARS: 0.12 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews PROFESSIONAL: 0.12 OTHER: 0.0 (H (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00225-05 LI TITLE OF PROJECT {80 characters or less. Title must St on one line between the borders.) Post-Inflamraatory Complications in Uveitis PR\NC\PAL\N\/SSJlGATOR(Ustotherprotessionalpersonnelbelow the Principal Investigator.) (Name, tideJaboratory.andinstitutBalfiliation.) PI: Chi-ChaoChan M.D. Medical Officer LI,NEI Others: Robert B. Nussenblatt Rachel R Caspi M.D. Clinical Director NEI Ph.D. Visiting Associate LI, NEI COOPERATING UNITS (if any) Bascom Palmer Eye Institute (Donald G. Pxiro, M.D.); University of Montreal, School of Medicine (Francois G. Roberge, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immimoregulation INSTITUTE AND LOCATION NEI, NIH, Bethesda. MD 20892 TOTAL MAN- YEARS: 0.26 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors D (a2) Interviews PROFESSIONAL: 0.26 OTHER: 0.0 1X1 (b) IHuman tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Post inflam matory complications in uveitis patients include destruction of photoreceptors, ghosis, choroidal scar, and formations of cyclitic and pupillary membranes, snowbanking and epiretinal membrane. Postinflammatory membrane composition may play an important role in the cause of these complications associated with uveitis. In this study, eyes enucleated from patients with end stages of chronic anterior uveitis (formation of cycUtic and pupillary mem- branes) and posterior uveitis (formation of cyclitic and epiretinal membranes) were evaluated immunohistochemically. Glial cells and proliferating Miiller cells were the major cellular components in these membranes. Basement membrane-like components and new collagens were the major extracellular membrane components. In vitro studies of the Miiller cell may help to elucidate the mechanism of action in its involvement of uveoretinitis. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00226-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (80 characters or less. Title must lit on one line between the borders.) Immunopathology of Ocular Onchocerciasis and Other Parasitic Diseases PRINCIPAL INVESTIGATOR (List other prolessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Chi-ChaoChan M.D. Medical Officer LI,NEI Others: Robert B. Nussenblatt M.D. Clinical Director NEI COOPERATING UNITS (it any) National Institute of AUergy and Infectious Diseases, Clinical Parasitic Diseases Section (Eric A. Ottesen, M.D.); World Health Organization (K. Awadzi, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. Nm. Bethesda. MP 20892 TOTAL MAN-YEARS: 0.33 PROFESSIONAL: 0.33 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors n (a2) Inten/iews Kl (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00241-04 LI TITLE OF PROJECT ( 80 characters or less. Tide must nt on one line between the borders.) Immiinopathology of Oailar Diseases in Htunans PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute alfiliation.) PI: Chi-Chao Chan M.D. Medical Officer LI,NEI Others: Robert B. Nussenblatt M.D. MingNi M.D. Marc de Smet M.D. Benjamin Rubin M.D. Clinical Director NEI Visiting Fellow LI, NEI Visiting Associate LI, NEI Senior Staff Fellow LI, NEI COOPERATING UNITS (if any) University of Minnesota, Department of Ophthalmology (Edward J. Holland, M.D.); Institute of Ophthalmology, University of London (Susan Lightman, M.D.); Lliopital de la Piti6, Paris, France (Phuc LeHoang, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH, Bethesda. MD 20892 TOTAL MAN-YEARS: 0.82 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors D (a2) Interviews PROFESSIONAL: 0.82 OTHER: 0.0 1X1 (b) IHuman tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Specimens from human ocular tissues with various diseases, such as uveitis, conjimctival and corneal diseases, metabolic genetic diseases, and tumors were studied using im- munoperoxidase technique as well as light and electron microscopic evaluation. In uveitis, immimocompetent cells and lymphokines are critical in the reflection of cHnical diagnosis, disease course, and prognosis. In non-uveitis conditions, alteration of cellular membrane surface markers and intracytoskeleton on the ocular resident ceUs may imply damage and abnormalities in these diseases. Elucidating the role of the relationships between infiltrating inflammatory cells and other cells in the clinical behavior of various diseases will increase our understanding of human ocular disorders. PHS 6040 (Rev. 1/84) GP0914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00264-01 LI PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Cytokines and Ocular Antigens in the Eye PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Chi-ChaoChan M.D. Medical Officer LI,NEI Others: Robert B. Nussenblatt IgalGery MingNi QuianLi M.D. Clinical Director NEI Ph.D. Head, Section on LI, NEI Experimental Immunology M.D. Visiting Fellow LI, NEI M.D. Visiting Fellow LI, NEI COOPERATING UNITS (if any) Laboratory of Immunoregulation, National Cancer Institute (Kouji Matsushima, M.D., Ph.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN- YEARS; 1.7 CHECK APPROPRIATE BOX(bS) D (a) Human subjects D (a1) Minors G (a2) Interviews PROFESSIONAL: 1.7 OTHER; 0.0 IS (b) Human tissues IXI(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Cytokines are communication signals between leukocytes and organ resident cells. Interleu- kin 1 (IL-1) a pleiotropic cytokine produced by many cell types, most notably macrophages, can stimulate a general inflammatory reaction by causing the activation of a variety of cells which then, among their other functions, release a cascade of other cytokines. One of them is Interleukin 8 (IL-8), which has more direct action of activating the neutrophils and T lympho- cytes. Intravitreal injection of IL-8 was compared to that of IL-1 in both Lewis and Fischer rats. IL-8 induced milder inflammation than IL-1. Lewis rats show a higher inflammatory response to the injections than do Fischer rats. These data suggest that IL-8 may be active as one component in neutrophil-mediated ocular disease. Further understanding of the role of each cytokine in ocidar inflammation, will require study of the production of endogenous cytokines in the eye. Ocular antigens play an important role in autoimmune diseases. S-antigen (S-Ag), a retinal soluble antigen, can induce experimental autoimmune uveoretinitis. In situ, study of S-Ag expression in human fetal eyes demonstrated an embryonic development pattern similar to that of other mammals. The expression of S-Ag in RNA of non-retinal fetal tissues may suggest its involvement in certain ocular diseases. Lens crystaUin proteins a, p and y were also foimd in human fetal iris and retinoblastoma cells. The roles of their expression in non-lenticvdar tissue require further investigation. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1. 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00251-03 LMDB TITLE OF PROJECT {80 charaaars or less. We must fit on one line between the borders.) ~ Regulation of the oA-Crystallin Promoter and Its Use for Genetically Engineering the Lens PRINCIPAL INVESTIGATOR (List other professional personnel behw the Principal Investigator.) (Name, title, laboramry, andinstitute affiliation.) PI: Others: Ana B. Chepelinsky Ph,D. Research Biologist Teresa I. Limjoco M.D. Jorge M. Sztein D.V.M. Graeme J. Wistow Ph.D. Visiting Fellow Visiting Fellow Visiting Scientist LMDB, NEI LMDB,NEI LMDB, NEI LMDB, NEI COOPERATING UNITS (if any) Gerontological Research Unit, Natl. Inst, of Health and Medical Research, Paris (Yves Courtois, Ph.D., Maryvonne Laurent, Ph.D.); Imperial Cancer Research Fund, London (Clive Dickson, Ph.D., Susan Jamie- son, Ph.D.); Lab. of Neurobiology, Natl. Inst, of Neurological Diseases and Stroke (Carolyn A. Bondy, M.D.) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section on Molecular Genetics INSTITUTE AND LOCATION NEI. NIH, Bethesda. MD 20892 TOTAL MAN- YEARS: 2.3 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews PROFESSIONAL: 2.3 OTHER: 0.0 n (b) Human tissues ISI(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The cis regulatory elements of the (xA-crystallin promoter responsible for the lens-specific expression of this gene and for its developmental regulation were further characterized. The results indicated that sequence -88 to +46 of the murine oA-crystallin gene contains the cis regulatory elements for lens-specific expression and for correct developmental regvdation of this gene in vivo. The ocA-crystaUin promoter (-366/+46) which has become a very usefvd tool to target gene expression to the lens, is being used to study how foreign gene expression in the lens affects the phenotype of the lens or the rest of the eye. Aldose reductase mRNA levels were foimd to be significantly increased in the lens of a transgenic mouse hne that carries the oA-crystallin promoter fused to the SV40 large T antigen. That mouse line develops a complete cataract as early as embryonic day 14. Several fusion genes containing the oA-crystalhn promoter fused to growth factors, oncogenes, interferon, or ribozymes are currently being used to study the effect of their expression in the eye. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00253-02 LMDB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Titia must fit on one line between the borders.) Regulation of Expression of Lens Fiber Membrane Genes PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratoiy, and institute affiliation.) PI: Ana B. Chepelinsky Ph.D. Research Biologist LMDB,NEI Others: M. Michele Pisano Teresa I. Limjoco GabrielaM.Tobal Ph.D. Staff FeUow LMDB, NEI M.D. Visiting Fellow LMDB, NEI Summer Student LMDB, NEI COOPERATING UNITS (if any) Harvard Medical School (David Paul, Ph.D.) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section on Molecular Genetics INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 1.8 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews PROFESSIONAL: 1.8 OTHER: 0.0 IS (b) Human tissues n(c) Neittier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project studies the regulation of expression of genes encoding lens fiber membrane proteins that may be involved in cell-cell commimication. Particular emphasis is put on the study of major intrinsic protein (MIP). We have cloned the MIP gene and have begun to analyze the cis regulatory elements responsible for its lens-specific expression. A recombinant clone containing the human MIP gene was isolated from a human leukocsrte genomic library using a bovine MIP cDNA (Gorin et al., Cell 39:49, 1984). This is the first lens-specific non-crystallin gene that has been cloned. Sequencing data indicates that this gene is 3.6 kbp long and contains four exons. The 5' flanking sequence of the gene contains a TATA box, two CCAAT boxes, and potential binding sites for the transcription factors Spl, NF-kB, and glucocorticoid receptor. Alu repeats are present in the 5' flanking region and third intron of the gene. In order to characterize the elements regulating expression of the MIP gene, 5' flanking DNA firagments of different lengths, synthesized by the polymerase chain reaction, were introduced upstream of the chloramphenicol acetyltransferase (CAT) reporter gene. Transfection of these gene constructs into explanted embyonic chicken lens epitheha and analysis of transient gene expression indicated that 253 bp of 5' flanking sequence contains an active promoter. The hximan MIP gene promoter functions with approximately the same efficiency as the promoter for the mouse y2 crystallin gene (also expressed specifically in lens fibers ). We are presently mapping the various cis regulatory elements responsible for the discrete temporal and tissue-specific expression of the MIP gene during lens fiber cell differentiation. PHS 6040 (Rev. 1/84) GPO 914-91 B DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00254-01 LMDB PERIOD COVERED October 1, 1989 to September 30. 1990 TfTLE OF PROJECT {80 characters or less. We must lit on one line between the borders.) Regfulatory Elements of the Opsin Promoter PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Ana B. Chepelinsky Ph.D. Research Biologist LMDB,NEI Others: Teresa I. Limjoco M.D. Visiting FeUow LMDB, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section of Molecular Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.35 PROFESSIONAL: 0.35 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors n (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00259-01 LMDB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (80 characters or less. Title must fit on one line tietween the borders.) Molecular Biology of the Cornea PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: R. Andrew Cuthbertson M.B.B.S., Visiting Associate LMDB.NEI Others: Joram Piatigorsky Ph.D. Ph.D. Chief LMDB, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section on Molecular Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda. MD 20892 TOTAL MAN- YEARS: 0.8 PROFESSIONAL: 0.8 OTHER: 0.0 CHECK APPROPRIATE BOX{ES) n (a) Human subjects n (a1) Minors n (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The cornea depends on a high degree of cellular differentiation to fulfill its unique fiinctions as an optically clear barrier and refiractive surface at the front of the eye. The deceptively simple overall structure of the corneal epithehum, stroma, and endothelium is ultimately dictated by the spatial and temporal regulation of gene expression in these cells. The molecular events that control phenotypic expression in the layers of the cornea are very poorly understood; we have therefore undertaken to isolate and characterize cDNAs that are preferentially (and perhaps uniquely) expressed in the corneal epithehum and endothehum, using subtractive hybridization and subsequent differential screening. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOIEY 00062-14 OGCS TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Irido-Comeal-Endothelial (ICE) Syndrome PR\HClPAL\HWEST\GMOR(Ust other professional personnel below the Principal Investigator.) (Narne, title, laboratory, and institute affiliation.) PI: Manuel B. Datiles M.D. Visiting Scientist OGCS.NEI Others: Muriel I. Kaiser-Kupfer M.D. Lessie McCain R.N. Chief Nurse Specialist OGCS, NET OGCS, NEI COOPERATING UNITS (if any) LAB/BRANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Cataract and Corneal Diseases INSTITUTE AND LOCATION NEL Nm, Bethesda. MP 20892 TOTAL MAN-YEARS: 0.15 CHECK APPROPRIATE BOX(ES) E (a) Human subjects D (a1) Minors D (32) Interviews PROFESSIONAL: 0.10 OTHER: 0.05 n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project was formerly titled "Progressive Essential Iris Atrophy." Patients with progres- sive essential iris atrophy with or without associated corneal disease are being recruited. Information is being gathered to evaluate the cUnical features and course of the disease process and to investigate aqueous hiomor dynamics in both affected and imaffected eyes. PHS 6040 (Rev. 1/84) GPOgi4-91B DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00187-07 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) The Effects of Corneal Contact Lenses on the Cornea PR\NC\PALimBSJ\GATOR(Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Manuel B. Datiles M.D. Visiting Scientist OGCS, NEI Others: Maria Elena Sibug M.D. Lessie McCain R.N. Juan Lopez M.D. Visiting FeUow Nurse Specialist Visiting Associate OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) LAB/BRANCH Ophthalmic Genetics and CUnical Services Branch SECTION Section on Cataract and Corneal Diseases INSTITUTE AND LOCATION NEI. NIH, Bethesda. MD 20892 TOTAL MAN-YEARS: 0.65 PROFESSIONAL: 0.40 OTHER: 0.25 CHECK APPROPRIATE BOX(ES) S (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues n(c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Short-term as well as long-term effects of contact lens wear on the cornea are being investigated. Changes ia corneal curvature, changes in corneal epithelial morphology, and changes in corneal endothelial cell morphology are being studied by specular microscopy. Analysis of the data obtained will help us vmderstand the dynamics involved in the interaction between a contact lens and the cornea, the risk to corneal tissues, and how a systemic or local disorder may increase these risks. Animal models showing abnormalities in corneal endotheUum similar to those in long-term contact lens wearers are also being explored. These are diabetic and galactosemic animal models. Treatment with aldose reductase inhibitors helps prevent these corneal abnormalities. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00188-07 OGCS PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Tide must fit on one line between the borders.) Documentation and Monitoring of Opacities in the Human Lens PRINCIPAL INVESTIGATOR (Ust other professional personnel below ttie Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Manuel B. Datiles M.D. Visiting Scientist OGCS,NEI Others: Rafael C. Caruso M.D. Benjamin Magno M.D. James Schumer M.D. Maria E. Sibug M.D. Lessie McCain R.N. Juan Lopez M.D. Visiting Scientist Visiting Associate StaffFeUow Visiting Scientist Clinical Technician Visiting Associate OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) Image Processing and Analysis Laboratory, Division of Computer Research and Technology, NIH (Banes Trus, Ph.D., Chief)- Chnical and Di^nostic Trials Section, National Cancer Institute, NIH (Sylvan Green, M.D.); Nuclear Medicine, Clinical Center, NIH (Joseph Frank, M.D.) LAB/BRANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Cataract and Corneal Diseases INSTITUTE AND LOCATION NEL NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 1.275 CHECK APPROPRIATE BOX(ES) ^ (a) Human subjects D (a1) Minors n (a2) Interviews PROFESSIONAL: 1.250 OTHER: 0.025 n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project uses different systems to develop objective and subjective methods to monitor and dociiment opacities in the human lens. We are actively recruiting patients with and vsdthout cataracts for reproducibility studies on the objective system — the Scheimpflug cameras (Zeiss and Topcon), retroillumination camera (Neitz), specular microscope (Keeler) and laser light-scat- tering spectroscope (Kowa). Tests of other systems will use sound (\iltrasonography) and nuclear magnetic resonance (magnetic resonance imaging). Ovir study of subjective systems or methods such as the effects of cataracts on visual perception, contrast sensitivity, and glare may be useful in identifying additional parameters for monitoring cataract presence, progression, or regression. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00212-05 OGCS TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between the borders.) Model Program for Collaboration Between Cataract Surgeons and Ophthalmic Researchers PR\UC\Pf^L\N\/ESJ\GAlOR(Ustotherprolessionalpersonnelbelowth0Principallnvestigator.)(Name, title, laboratory, and institute affiiiation.) PI: Manuel B. Datiles M.D. Visiting Scientist Others: D. James Schumer M.D. Benjamin Magno M.D. Juan Lopez M.D. Staff FeUow Visiting Associate Visiting Associate OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) LAB/BRANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Cataract and Corneal Diseases INSTITUTE AND LOCATION NEI, NIH, Bethesda. MD 20892 TOTAL MAN-YEARS: 0.95 PROFESSIONAL: 0.95 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) H (a) Human subjects n (al) Minors D (a2) Interviews n (b) Human tissues n(c) Neitlier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) There is an extreme scarcity of human cataract material because of an abrupt shift of cataract surgical technique from intracapsular (intact lens) to extracapsular (fragmented lens), primarily because of advent of the use of intraocular lens. We are exploring ways by which fragmented lens materials can be maximally used in cataract basic research through close collaboration with cataract surgeons and basic researchers and through modification of techniques by both groups. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00249-02 LI TfTLE OF PROJECT {80 characters or less. Title must fit on one line between ttie borders.) Cytokines in Human Intraociilar Fuids Pn\NC\PAL\H\/ESJ\GAJOH (LJst other professional personnel below the Prindpal investigator.) (Naine.ddB, laboratory, and institute affiliation.) PI: Janet L. Davis M.D. Senior Staff Fellow LI,NEI Others: Robert B. Nussenblatt M.D. Clinical Director NEI COOPERATING UNITS (if any) Eye Research Institute, Boston, MA (Alex E. Jalkh, M.D., Charles Schepens, M.D.); Univer- sity of Miami, Miami, FL (Harry W. Flynn, Jr., M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immimoregulation INSTITUTE AND LOCATION NET, NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 0.32 CHECK APPROPRIATE BOX(ES) □ (a) Human subjects n (a1) Minors D (32) Interviews PROFESSIONAL: 0.32 OTHER: 0.0 IXI (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00065-13 IRP PERIOD COVERED October 1. 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Physiological Studies of the Primate Visual System PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Francisco M. de Monasterio M.D., D.Sc. Medical Officer IRP, NEI COOPERATING UNITS (if any) LAB/BRANCH Office of the Director of Intramural Research SECTION INSTIUTTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 0.3 PROFESSIONAL: 0.3 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (b) Human tissues IS (c) Neither n (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project involves the study of the physiological organization of neurons of the visual system of primates, with emphasis on the chromatic properties of color-opponent ganglion cells and of cells from the lateral geniculate nucleus and the primary visual cortex of macaques. PHS 6040 (Rev. 1/84) GPO 91'*-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00122-10 IRP PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 characters or less. Title must fit on one line between the borders.) Anatomical Studies of the Primate Visual System PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Francisco M. de Monasterio M.D., D.Sc. Medical Officer IRP, NEI COOPERATING UNITS (If any) LAB/B RANCH Office of the Director of Intramural Research SECTION INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.7 PROFESSIONAL: 0.7 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects Kl (b) Human tissues D (c) Neither D (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project involves the study of the anatomical properties and organization of cells in the visual system of primates, with emphasis on the retina and the visual cortex. The studies include (1) the electroretinogram (ERG) mediated by blue-sensitive cone signals before and after the staining of putative blue-sensitive cones by tissue-reactive dyes, (2) the distribution patterns of cones in the hujnan donor retinas with a diagnosis of diabetic retinopathy, and (3) the anatomical association of outer-retinal cells selectively stained with tissue-reactive dyes. (1) Evaluation of the spectral properties of the macaque monkey cones selectively stained with tissue-reactive dyes, which have been tentatively identified as the "blue" cones, has been hindered by the fact that the dye itself alters the spectral transmission of the ocular media. The study of ERG sensitivity of the same animals before and after the selective labeling of the putative blue cones with the clear monochlorotriazinyl reagent Sandospace S, which can be marked with a secondary dye, has continued. This reagent produced a sharp reduction of the test and field sensitivity of the ERG b-wave mediated by blue cone signals, providing further evidence that the above tentative identification is correct. (2) Evidence of a cone population vsdth a point pattern resembling that of cones selectively stained by tissue-reactive dyes was obtained in initial studies of the retinas of diabetic human donors. Although such dyes were not injected into the eyes of these donors, in vitro staining with more conventional dyes showed a differential receptor labeling, in which cones with the pattern mentioned above were fovmd to be more densely stained than other cones. (3) A systematic Ught-microscope study of the anatomical association of blue cones and horizontal and bipolar cells that have been selectively stained by several tissue-reactive dyes has been initiated in the macaque retina. These studies provide information on the probable retinal circuitry of the blue-sensitive cone pathway of primate retina. PHS 6040 (Rev. 1 1SA) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00218-05 LI TITLE OF PROJECT {80 characters or less. Title must fit on one iine between the borders.) Acquired Immune Deficiency Syndrome PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Marc D. de Smet M.D. Visiting Scientist LI, NEI Others: Robert B. Nussenblatt M.D. Benjamin I. Rubin M.D. Scott Whitcup M.D. Rubens Belfort M.D. Juan Lopez M.D. Clinical Director NEI Senior Staff FeUow LI, NEI Senior StafFFeUow LI, NEI Visiting Scientist LI, NEI Visiting Fellow LI, NEI COOPERATING UNITS (if any) Lab. of Cellular and Molecular Biology, Natl. Cancer Inst. (Dharam Ablashi, D. V.M.); Dept. of Critical Care Medicine, Clinical Center (Henry Masur, M.Dj; Lab. of Tumor Cell Biology, Natl. Cancer Inst. (Robert C. Gallo, M.D.); Lab. of Immunoregulation, Natl. Inst, of Allergy and Infectious Diseases (H. Clifford Lane, M.D.); Natl. Inst, of Allergy and Infectious Diseases (Anthony Fauci, M.T).); Pediatric Branch, Natl. Cancer Inst. (Phil A. Pizzo, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immimoregulation INSTITUTE AND LOCATION NEI, Nm, Bethesda, MD 20892 TOTAL MAN-YEARS: 1.2 PROFESSIONAL: L2 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) S (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Patients with AIDS are at risk of developing significant ocvdar problems, some of which can lead to blindness. Left vintreated, CMV retinitis is a major cause of blindness in AIDS patients. We have looked at the effect of the new antiviral drug foscarnet and determined its ability to inhibit progression of the virus. We have also started investigating the usefulness of adjunctive therapy with laser photocoagulation in preventing progression of the disease. In the course of oiu" studies, we have also identified new manifestations of known infectious agents in patients with AIDS, namely, HZV retinitis. In conjtLuction with the Pediatric Branch of the National Cancer Institute, we have been following about 125 children with symptomatic HIV to determine the nat\iral history of the disease and to determine the probabihty of significant ocular disease in these children. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00266-01 LI TITLE OF PROJECT {80 characters or less. Titte must fit on one line between the txirders.) Characterization of Immune Responses to S-antigen PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tiUa, laboratory, and institute affiliation.) PI: Marc D. de Smet M.D. Visiting Scientist LI,NEI Other: Igal Gery Robert B. Nussenblatt Scott Whitcup Xiao Yan Zhang Ph.D. Head, Section on LI, NEI Experimental Immunology M.D. Clinical Director NEI M.D. Senior Staff Fellow LI,NEI M.D. Guest Worker LI,NEI COOPERATING UNITS (if any) Department of Ophthalmology, Kurume Univ., Japan (M. Mochizuki, M. D.); INSERM-86, Laboratory of Ocular Immunopathology, Paris (J-P Faure, Ph.D.); Hadassah Hospital, Jeru- salem, Israel. LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immiinoregulation INSTITUTE AND LOCATION NEL Nm, Bethesda. MD 20892 TOTAL MAN-YEARS: 0.44 CHECK APPROPRIATE BOX{ES) IS (a) Human subjects D (a1) Minors D (a2) Inten/iews PROFESSIONAL: 0.44 OTHER: 0.0 1X1 (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Experimental autoimmune uveitis (EAU) is a disease model induced in genetically suscepti- ble animals by injection of various soluble retinal antigens in adjuvant. The best characterized of these antigens is S-Ag. The uveitis it induces is critically dependent on the presence of activated T cells. It has been established that T cells become activated as a result of their interaction with primed antigen-presenting cells, a process through which the immvmogenic protein is internal- ized and partially digested. Fragments from the original protein are then reexpressed on the cell siu-face where they can interact with T cells. Workis being carried out to identify and characterize those fragments which are responsible for the chnical response in animals as well as in humans. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00267-01 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (80 charaaers or /ess. TWa must fit on one line between the borders.) Modiilation of Immune Functions Using the Immunotoxin IL2-PE40 PR{NClPAL\N\/ESJ\GfiiJOR (Usi other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Marc D. de Smet M.D. Visiting Scientist LI,NEI Others: Stefan S. Thurau Carl P. Herbert Robert B. Nussenblatt M.D. Special Volunteer LI.NEI M.D. Special Volunteer LI,NEI M.D. Clinical Director NEI COOPERATING UNITS (if any) Laboratory of Molecular Biology, National Cancer Institute (Ira Pastan, M.D.); University of Montreal, School of Medicine (Francois G. Roberge, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL MAN-YEARS; 0.6 PROFESSIONAL: 0.6 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues K](c) Neitlner SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) IL-2-PE40 is a recombinant chimeric protein composed of interleukin-2 (IL-2) and ofPseu- domonas endotoxin (PE40), minus its ceU-binding domain. This toxin, once internalized, kills cells by irreversibly inhibiting protein synthesis. Hence, cells bearing ILr2 receptors on their surface, which are prime targets for this toxin, can be effectively removed from an organism without producing too many iintoward side effects. In experimental autoimmvme uveitis (EAU), T cells play a major role. Once activated, these T cells express on their cell surface IL-2 receptors, which can be targeted by the chimeric toxin and ehminated. We have demonstrated that this toxin is able to reduce significantly the incidence and severity of experimentally induced uveitis. IL-2PE40 is also able to reduce the incidence and severity of corneal graft rejection following systemic administration of the toxin. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PRCXIECT NUMBER ZOl EY 00231-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must lit on one line tietween the borders.) Cell Surface Antigens on Retinoblastoma Cells PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Barbara Detrick Ph.D. Expert LI,NEI Others: John J. Hooks Ph.D. Gerald J. Chader Ph.D. C aroHne Percopo B . S. Head, Section on Immunology and Virology Chief Biologist LI.NEI LRCMB, NEI LI,NEI COOPERATING UNITS (if any) Tximor Biology Section, Laboratory of Biology, National Cancer Institute (Charles Evans, M.D.); Walter Reed Army Medical Center (Norman Katz, M.D.); University of Maryland, Bal- timore (MeryLa Rodrigues, M.D.) LAB« RANCH Laboratory of Immunology SECTION Section on Immimoregulation INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 0.6 PROFESSIONAL: 0.4 CHECK APPROPRIATE BOX(ES) IS (a) Human subjects n (a1) Minors n (a2) Interviews OTHER: 0.2 D (b) Human tissues n(c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00235-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 charactars or less. We must St on one line between the borders.) Tdentification and Modiilation of Class II Antigens PRINCIPAL INVESTIGATOR ( Ust other professional personnel below the PrindpaJ Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Barbara Detrick Ph.D. Expert LI,NEI Others: John J. Hooks Chi-Chao Chan CaroUne Percopo Robert B. Niissenblatt Ph.D. Head, Section on LI, NEI Immunology and Virology M.D. Medical Officer LI, NEI B.S. Biologist LI, NEI M.D. Clinical Director NEI COOPERATING UNfTS (if any) University of Pennsylvania (G. Aguirre, D.D.S., Ph.D.); Duke University (Barton F. Haynes, M.D.); Paris, France (Laurence Boxomsell, M.D.). LAB« RANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 0.44 CHECK APPHOPRIA 1 1 BUX(bb) D (a) Human subjects n (a1) Minors D (a2) Interviews PROFESSIONAL: 0.34 OTHER: 0.10 n (b) Human tissues ia(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/S4) QP0 9U-91i DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00261-01 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TfTLE OF PROJECT [80 characters or less. Title must lit ort one line between the borders.) Identification of Pathogens in Ocxilar Tissues by PCR PR\NC\PAL\ti\/€ST\GATOR (List other prolessional personnel below the Principal Investigator.) (Name, tide, laboratory, and instituteaffiliation.) PI: Charles E. Egwuagu M.P.H., Scientist LI,NEI Others: Antoine P. Brezin Robert B. Nussenblatt Rubens Belfort, Jr. Rashid Mahdi Ph.D. M.D. M.D. M.D. Visiting Fellow LI, NEI Clinical Director LI, NEI Visiting Scientist LI, NEI Biologist LI, NEI COOPERATING UNITS (if any) stitute of Allergy and Infedbious Diseases (Ricardo T. Gazzinelli, Ph.D.) Brazil- , stry of ational In- LAB/B RANCH Laboratory of Immunology SECTION Section of Experimental Immunology INSTITUTE AND LOCATION NEI. Nm. Bethesda, MP 20892 TOTAL MAN-YEARS: 2.0 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 1.9 OTHER: 0.1 [XI (b) Human tissues n(c) Neither n (a) Human subjects n (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) We have used the polymerase chain reaction (PCR) to amplify fi-agments specific to Tox- oplasma gondii firom two cases of ocular toxoplasmosis. In both cases, PCR revealed the presence of toxoplasmic DNA although optical microscopy showed toxoplasmic cysts in only one eye. Our success in identifjdng the parasite in these samples indicates the usefulness of this technique as an adjunct to ciurently available diagnostic methods. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00262-01 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TFTLE OF PROJECT {80 characters or less. Title must fit on one line between the tiorders.) TCR Gene Usage in Experimental Autoimmiine Uveoretinitis (EAU) PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Charles E. Egwuagu M.P.H.,Ph.D. Scientist LI.NEI Others: Igal Gery Robert B. Nussenblatt Christopher Chow Evelyne Beraud Rachel Caspi Rashid Mandi Antoine P. Brezin Ph.D. M.D. B.A. Ph.D Ph.D. M.D. Head, Section on LI, NET Experimental Immunology Clinical Director LI, NEI HHMI-NIH Scholar U, NEI Visiting Associate LI, NEI Visiting Associate LI, NEI Biologist LIINEI Visiting Fellow COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section of Experimental Immunology INSTITUTE AND LOCATION NEI, NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: L4 PROFESSIONAL: LI OTHER: 0.3 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors n (a2) Interviews D (b) Human tissues IXl(c) Neitlier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project is aimed at learning about the T cell receptors (TCR) expressed by T cells that mediate a group of inflammatory eye diseases referred to as "uveitis". Most emphasis in FY 1990 has been on the isolation and characterization of the genes that code for these surface proteins used by T cells for antigen recognition. We have investigated the TCRs expressed on T cell hnes and clones specific for S-antigen and interphotoreceptor retinoid-binding protein (IRBP) and compared V-region gene usage between T cells capable of transferring EAU (uveitogenic) and non-pathogenic T cells. Our analyzsis of the VP8 gene locus for genomic rearrangements demonstrated the predominant VpS gene rearrangement, even in oiir most pathogenic T cell lines, suggesting that small proportions of the cells in our T cell lines that induce EAU use the V|38 gene element. On the other hand, our detection of Vp8 transcripts in uveitogenic T ceU lines and the level of VpS gene expression coiild be correlated with the abiHties of our various T cell lines and clones to induce EAU, which suggests that T cells expressing the Vp8 phenotype may be involved in the etiology of EAU. However, Northern analysis lising a probe specific to Vp8.2 revealed that, unlike the pathogenic T cells involved in other experimental autoimmune diseases, uveitogenic T ceU hnes express a member of the VpS TCR family that appears to be similar to but distinct fi-om Vp8.2. We have cloned and sequenced the VP8.2-like cDNAs derived fi-om S-antigen and IRBP-specific uveitoge- nic T cell Hnes. The gene Smm. the IRBP line shows approximately 99% sequence homology within rat VP8.2, while the gene fix)m the S-antigen Hne is only 90% homologous to Vp8.2. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00189-07 LMOD TITLE OF PROJECT (80 characters or less. 7We must fit on one line between the borders.) Oxidation of Proteins in Cataractogenesis Pf^tfiC\PAUmESJ\GAJOR(Ustoiher professional personnel below the Prindpal Investigator.) (Name, titlejaboratory, and Institute PI: Donita L. Garland Ph.D. Research Chemist LMOD, NEI Others: Jose Jimenez Ph.D. Lorenzo Merola M.S. Visiting Fellow Chemist LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Disease s SECTION ' Section on Cataracts INSTITUTE AND LOCATION NEL Nm. Bethesda. MD 20892 TOTAL MAN- YEARS: 3.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors a (a2) Interviews PROFESSIONAL: 3.0 OTHER: 0.0 D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Oxidative changes of lens proteins are thought to occur with aging and to contribute to the development of cataracts. The goals of this project are to determine (1) the extent of oxidative modification of crystallias and metabolic enzymes in both normal and cataractous lenses, (2) the nature of the modifications and mechanisms leading to the changes, and (3) the effect of the modifications on the structure and function of lens proteins. Bovine and rat lenses are used. The approach is to study the modifications of lens proteins afl;er treatment in vitro by metal-cat- alyzed oxidation systems. Structviral alterations induced by these oxidative systems were examined by circular dichro- ism and peptide mapping. Trace metal analysis of bovine aqueous and rat and bovine lenses indicated that copper and iron are both present in micromolar concentrations. Further studies on fetal bovine lenses demonstrated that copper and iron are not associated with any crystalhn to an appreciable extent in vivo. In vitro, copper appears to interact specifically with some of the bovine gamma-crystalUns and at greater than stoichiometric levels induces protein aggregation. These results support the possibility that metal-catalyzed oxidative reactions may contribute to age-related changes in lens. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00069-13 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. We must tit on one line between the borders.) Inimiine Responses to Ocular Antigens PB]HC\PM\NVES'i\GA\OR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Igal Gery Ph.D. Head, Section on Experimental Immunology LI, NEI Others: Satoshi Kotake M.D. Yoichi Kawano M.D. Yoichi Sasamoto M.D. Xiao-Yan Zhang M.D. William Lipham B A. Barbara Vistica B A. Gerald J. Chader Ph.D. Barbara Wijsgert Ph.D. T. Michael Redmond Ph.D. Visiting Fellow Visiting Fellow Visiting Fellow Guest Worker HHMI-NIH Scholar Microbiologist Chief Head, Section on Biochemistry Senior Staff Fellow LI, NEI LI NEI LI, NEI LI NEI LI, NEI LLNEI LRCMB, NEI LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) Metabolism Branch, Division of Cancer Biology and Diagnosis, National Cancer Institute (Jay A. Berzofsky, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Experimental Immunology INSTITUTE AND LOCATION NEI. NIH. Bethesda. MD 20892 TOTAL MAN-YEARS: 3.2 CHECK APPROPRIATE BOX(ES) n (a) Human subjects □ (a1) Minors D (a2) Interviews PROFESSIONAL: 3.0 OTHER: 0.2 D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project is aimed at learning about the pathogenesis of inflammatory eye diseases grouped under the term "uveitis." Our effort in FY1990 focused on studies in experimental animals and has yielded the following main achievements: 1. The active site of the immunodominant and highly uveitogenic determinant of bovine Lnter- photoreceptor retinoid-binding protein (IRBP) was localized to a nonapeptide at sequence 1182-1190. Four of the amino acids of this peptide were identified to be pivotal to its immunological activities; residues 1182 and 1190 are essential for its binding to the antigen presenting ceU while 1188 and 1189 interact vnth. the T cell receptor. 2. Lymphocytes become highly uveitogenic by their activation in culture. Data collected this period show dissociation between the generation of uveitogenicity and the proliferation of activated lymphocj^es. 3. Concanavalin A (Con A), which stimulates vigorous proliferation responses in spleen (Sp) and lymph node (LN) cell cultures, also generates uveitogenicity in Sp cells, but not in LN cells. This lack of Con A effect in LN cvdtures was foimd to result from a deficiency in an accessory cell population present in the Sp. 4. Lymphocytes sensitized to non-dominant IRBP peptides do not recognize whole IRBP in culture as they do in the eye when mediating uveitis in rats. However, these lymphocytes do recognize IRBP following its digestion by endopeptidases. This finding provides an explanation for the aforemen- tioned discrepancy and suggests that the recognition of IRBP in the eye is facilitated by its cleavage by tissue enzymes. 5. Rats injected with the uveitogenic peptide 1177-1191 in aqueous solution were found to be resistant to EAU induction by this peptide when it was injected in its uveitogenic form, in adjuvant emulsion. Furthermore, the treated rats were also resistant to induction of EAU by whole IRBP. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00049-12 LSR PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 characiers or less. Tille must fit on one line between the borders.) Cerebral Cortical Mechanisms for Eye Movements and Visual Attention PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and Institute affiliation.) PI: Michael E. Goldberg M.D. Chief, NMS LSR,NEI Others: Edmond J. FitzGibbon Carol L. Colby Jean-Rene Duhamel Carl R. Olsen Suzanne Y. Musil Edward L. Keller M.D. Senior Staff FeUow LSR,NEI Ph,D. Senior Staff FeUow LSR,NEI Ph.D. Visiting Scientist LSR.NEI Ph.D. Guest Researcher LSR,NEI Ph.D. NRSA Fellow LSR,NEI Ph.D. Guest Researcher LSR, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Section on Neuro-Ophthalmologic Mechanisms INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN- YEARS: 5.6 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 4.5 OTHER: 1.1 13(0) Neither n (a) Human subjects D (b) Human tissues n (a1) Minors D (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Two different lines of inquiry were followed to determine how the cerebral cortex and its efferent regions control eye movements and visuospatial attention. In one, activity of movement neurons of the superior coUiculus was studied in relation to saccades evoked by electrical stimidation of either the frontal eye field or the superior colhculus and modified by their temporal proximity to visually evoked saccades. In the other, visual neurons in the posterior parietal cortex were studied using double-step tasks to see how this cortex might maintain spatial accuracy when there was a dissonance between the retinal location of a stimulus and the saccade necessary to acquire that stimvdus. Neurons in this region discharged when the monkey made a saccade of the proper direction to acquire a stimixlus, whether or not that stimiilus lay in the neiiron's receptive field as studied in a routine fixation task. Such neurons required the presence of a visual stimulus, suggesting that in the posterior parietal cortex, spatial acciuracy is maintained by coordinate transformation of a visual map rather than by the explicit coding of target position in space. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00246-03 OGCS TITLE OF PROJECT {80 characters or lass. Title must tit on one line between the borders.) Molecular Genetics of Retinal Degenerations PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Michael B. Gorin M.D.,Ph.D. Medical Officer OGCS,NEI Others: Steven Snyder M.D. Tatiana PutiHna Ph.D. Staff FeUow Special Volunteer OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) Retinitis Pigmentosa Foundation: Northwestern Univ. (Larry Rnto, Ph.D.): Univ. of Linkopine, Linkoping, Sweden (Knstina Narfstrom, V.M.D.); Dept. of Biochemistry, UCLA (David Sigman, Ph.D. J; Lab. of Molecu- lar Microbiology, Natl. Inst, of Allergy and Infectious Diseases (Christine Kozak, Ph.D.) LAB/BRANCH Ophthalmic Genetics and Chnical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda. MD 20892 TOTAL MAN-YEARS: 1.3 PROFESSIONAL: L3 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The pvirpose of this project is to identify the genes responsible for different inherited retinal disorders in animal models and to estabhsh the genetic relationship of these animal disorders to forms of human retinal degenerations and conditions. "Reverse" genetic approaches are being appUed to specific animal models of retinal dysfunc- tion, including new methods for cloning regions associated with a mapped genetic disorder. Polymerase chain ampHfication methods are being used to evaluate interspecies differences in specific genetic transcripts. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00135-18 IRP PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (flO characters or less. Title must fit on one line between the borders.) Biochemistry of Retina and Pigmented Epithelivun in Health and Disease PR\HC\PAL\N\/BSl\GAJOR (Ust other professional personnel below ttie Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Helen H. Hess M.D. Medical Officer (Research) IRP,NEI COOPERATING UNITS (if any) LAB/BRANCH Office of the Director of Intramural Research SECTION INSTITUTE AND LOCATION NEI. NM. Bethesda, MD 20892 TOTAL ^MN-YEARS: 1.0 PROFESSIONAL: 1.0 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (b) Human tissues Kl (c) Neither n (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The effects of nutrition, oxidation, and other environmental factors (light intensity or darkness) on incidence and progress of posterior subcapsular opacities (PSO) associated with retinal degeneration are being studied in pink-eyed Royal College of Surgeons (RCS) rats, in which rod photoreceptor outer segment debris acciunulates secondary to a phagocytic defect in retinal pigmented epitheHum. Evidence was obtained that oxidative changes in polyimsatxirated fatty acids in debris led to water-soluble toxic aldehydes, detectable in the vitreous and toxic to lens cells and membranes. Dystrophic rats fed a natural ingredient diet (NIH-07) are highly susceptable to retinal light damage, beginning at 1-4 footcandles (fc) intensity: 27% of the rats develop mature cataracts by 7-12 months. Increased light intensity (cycUc or constant) increased the percentage of rats with matiire cataracts, while dark-rearing from birth prevented PSO and matiu-e cataracts. A purified diet (AIN-76A), fortffied with 0.4% p-carotene + 0.01% BHT, also prevented PSO and mature cataracts. Rhodopsin bleaching appears to be essential for retinal light damage and PSO. A 100% incidence of bilateral mature cataracts occurred in dystrophic rats given 48 hours of 700-fc constant Hght between postnatal days 22 and 28, when rhodopsin is increased 70% in debris. A similar incidence of bilateral cataracts occxirred in congenic control RCS rats given 18 days of dark adaptation to increase rhodopsin by 50%, followed by the same constant light exposxure. In vitro, free retinaldehyde can act as a photosensitizer to generate singlet oxygen, an extremely energetic oxidant. Present resvilts suggest a similar effect in vivo, with damage to both lipids and proteins. Current studies are directed toward exploring how many days retinal degeneration can be delayed by different antioxidant-containing diets. Antioxidants may slow or prevent cataracts in some human retinal diseases. PHS 6040 (Rev. 1/84) GP0914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00232-05 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line t>etween the borders.) Interferon System in Cellxilar Function and Disease PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: John J, Hooks Others: CaroUne Percopo Christian Hamel Shirin Barer Gerald Chader Ph.D. Head, Section on Immunology and Virology LI,NEI M.S. Biologist LI,NEI M.D. Visiting Fellow LI,NEI M.D. LI,NEI Ph.D. Chief LRCMB, NEI COOPERATING UNITS (if any) New York University, School of Medicine (Jan VUcek, M.D.); Tumor Biology Section, Laboratory of Biology, Division of Cancer Etiology, National Cancer Institute (Charles Evans, MZ).}; Vaccine Research and Development Branch, Division of AIDS, National Institute of Allergy and Infectious Diseases (Barbara DetricK, Ph.DO LAB/BRANCH Laboratory of Immunology SECTION Section on Immimology and Virology INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL NMN- YEARS; LI CHECK APPROPRIATE BQX(hS) D (a) Human subjects D (a1) Minors n (a2) Interviews PROFESSIONAL: 0.7 OTHER: 0.4 D (b) Human tissues (Xl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The interferon (IFN) protein, which can modify a variety of biological activities, is considered one of the body's regulatory proteins. Numerous studies have indicated that the IFNs are potent immunoregulators. During the past year, we have been studying the ways in which IFN proteins interact with cells of the immune system and with cells in the ocular microenvironment. Using immunocytochemical analysis, we have developed a sensitive method of identifying the lymphokines IFN-y and interleukin 2 (IL-2) at the site of tissue damage. We have identified these lymphokines in inflammatory eye diseases. The presence of these lymphokines is associ- ated with lymphocyte infiltrate predominantly of T cell origin and with the expression of major histocompatibility complex (MHC) class II antigens on both the infiltrating cells and retinal pigment epithelial (RPE) cells. Experimentally, we have shown that this direct intravitreal inoculation of recombinant rat IFN-Y results in the expression of MHC Class II antigen (la) in a variety of ocular cells. In conjunction with la expression, two striking changes were noted: an iritis and infiltrating cells in the inner retinal layers. Both of these phenomena have been observed in certain inflamm atory eye diseases. IFN-Y is known to be a potent regulator of gene expression. We foxmd that IFN-Y enhances the expression of retinal S-antigen, a specific neuronal ceU marker. Preliminary studies indicate this IFN-Y is acting at the level of transcription. These observations indicate that IFN-y induced MHC class II antigen expression may serve as a local ampHfication system in autoimmune and inflammatory eye disease. A better under- standing of the role of lymphokines in the mechanisms involved in the development of autoim- munity and inflammation may be beneficial in developing treatments for these diseases. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00233-05 U PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or lass. Title must fit on one line between the borders.) Studies on the Bioregulatory Aspects of the Retinal Pigment Epithelial Cell PBit^C\PAl\N\IESJ\GAJOH(UstotherprofessionalpersonnBlbelowihePrincipallnvestigator.)(Nanie, title, laboratory, and institute affiliation.) PI: John J. Hooks Ph.D. Head, Section on LI, NET Others: CaroHne Percopo M.S. Laura Gaspers- Velu M.D. Shuji Suziiki M.D. Immunology and Virology Biologist LI, NEI Visiting Associate LMOD, NEI Visiting Associate LI, NEI COOPERATING UNITS (if any) Hopital St. Louis, Paris, Prance (Lawrence Boumsell, M.D.); Univ. of Nice, Prance (Alain Bernard, M.D.); NRED, NIH (Reuben Sigraganian, M.D.); Univ. of Virginia. Charlottesville, VA (Stanley A. Vinores, Ph.D., Peter Campochiaro, M.D.)jX)regpn Health Sciences Univ., Portland, OR (Stephen R. Planck, Ph.D., James T. Rosenbaum, M.D.); Division of AIDS, National Institute of Allergy and Infectious Diseases (Barbara Detrick, Ph.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immiinology and Virology INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN- YEARS: 1.96 PROFESSIONAL: 1.76 OTHER: 0.20 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors n (a2) Interviews IXI (b) l-iuman tissues n(c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The retinal pigment epithelial (RPE) ceU has a major regulatory function in the eye, in a variety of ways influencing the maintenance of retinal integrity and function. In order to study this cell more effectively in vivo and in vitro, we have produced monoclonal antibodies directed against human RPE cells. Using immttnoperoxidase assays (ABC), we have identified two mouse IgG monoclonal antibodies that react with the human RPE cell. The monoclonal antibodies are both specific for the RPE cell within the eye because they do not react with any other ocular structures. Moreover, these antibodies do not cross-react with human skin, kidney, or peripheral mononuclear cells. These antibodies recognize cell surface molecules that must be highly conserved, being foimd in man, monkey, rat, mouse, cow, chicken, and fi*og. These antibodies, which detect epitopes present solely on RPE cells, provide us with the xmique opportunity to evaluate a variety of aspects of RPE cell development and function. Studies of RPE cell development indicate tliat the epitopes appear only after the cells have begim terminal differentiation. Moreover, these studies indicate that a very specific product of the RPE cell is synthesized as the photoreceptor outer segment starts to develop, suggesting that this product could be involved in an essential step of the outer segment development. Studies on RPE migration also demonstrate the value of these antibodies in evaluating epiretinal membrane formation. These are the first monoclonal antibodies directed solely at the human RPE cell. Further characterization and studies of these antibodies should prove useful in the identification of RPE cells in situ and in vitro. This immunoglobulin wiU allow us to probe the bioregulatory functions of the cells. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00234-05 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT ( 80 characters or less. Title must fit on one line between the borders.) MHC Class II Antigens in the Pathogenesis of Inflammatory Diseases PRINCIPAL INVESTIGATOR (List other professional personnel belovi the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: John J. Hooks Ph.D. Head, Section on LI,NEI Other: Caroline Percopo Robert B. Nussenblatt Immunology and Virology M.S. Biologist LI,NEI M.D. Clinical Director NEI COOPERATING UNITS (if any) Vaccine Research and Development, Division of AIDS, National Institute of Allergy and In- fectious Diseases (Barbara Detrick, Ph.D.) UB/B RANCH Laboratory of Immunology SECTION Section on Immimology and Virology INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 0.4 PROFESSIONAL: 0.3 OTHER: 0.1 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SU MMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) MHC class II antigens, HLA-DR in the human and la in the mouse, are membrane-boiind glycoproteins encoded by genes of the major histocompatibility complex. Expression of these antigens is of great fiinctional importance for the initiation and perpetuation of immune responses. In a number of immunopathologic conditions, HLA-DR antigen-negative cells are stimulated to express class II antigens. In these cases, an immunologic role has been postulated for the class II antigen expression. We showed that cytokine-activated RPE cells express MHC class II antigens. These la-pos- itive cells can both process retinal antigens and present them to specifically sensitive T-helper lymphoctyes. These studies indicated that cytokine activated RPE cells may be a basic compo- nent of ocular immvmity. These studies on MHC class II antigen expression in localized autoimmune diseases provide evidence that the activation of these antigens may contribute to the immunopathogenesis of these diseases. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00240-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Tide must lit on one line between the borders.) Virus Infections in the Eye PR INCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: John J. Hooks Ph.D. Head, Section on LI, NEI Immunology and Virology Others: Susan Robbins Ph.D. Postdoctoral Fellow Christian Hamel M.D. Visiting Fellow CaroHne Percopo M.S. Biologist Charles Evans M.D., Ph.D. Medical Officer LI, NEI LI, NEI LI, NEI LB, NCI COOPERATING UNITS (if any) Div. of AIDS, Natl. Inst, of Allergy and Infectious Diseases; (Barbara Detrick, Ph.D.); Wilmer Eye Institute, Johns Hopkins Hospital, Baltimore (W.Tlichard Green); Dept. of Pathology, Uniformed Services Univ. for Health Sciences, Bethesda, MD (Katharine Holmes, Ph.D.): Dept. of Ophthalmology, Ruprecht-Karl's Univ., Heidelberg, Germany (Ellen Kraus-Mackiw, M.D.); Dept. of Ophthalmology, Univ. of Munich, Germany (Otto F. Scheiffarth, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunology and Virology INSTITUTE AND LOCATION NEL NIH, Bethesda, MD 20892 TOTAL MAN- YEARS: 1.0 PROFESSIONAL: 0.9 OTHER: 0.1 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors D (a2) Interviews IS (b) Human tissues D(c) Neither SU MMA RY OF WORK (Use standard unreduced type. Do not exceed the space provided.) During the past year, we have studied various virologic and immunopathologic processes that occur when viruses replicate in the ocular microenvironment. This project consists of three areas: (1) studies on coronavirus infection in ocular and optic nerve cells; (2) determination of the possible roles of viruses in human diseases; (3) antiviral therapeutic actions of cjrtokines and drugs. We have established that miirine coronavirus can induce ocular disease and thus may be used as a model system for studying retinal degenerative diseases. This model has many imique features. The virus is capable of inducing an acute infection in the presence of mild inflammation. The initial retinal damage is followed by clearance of the virus and progressive retinal destruc- tion, even months after the virus is gone. This disease may be considered a model for degener- ative diseases of the pigment epithelium and photoreceptors in humans. The need for effective drug treatment and prevention of herjjes virus and other viral diseases has assiimed growing importance. We foimd that leukoregulin, a naturally occiuring immuno- logic cytokine, increases the antiviral actions of the drug acyclovir. These findings which demonstrate that combination immunotherapy and chemotherapy can produce substantial inhibition of herpes virus replication, provide a rationale for the application of this approach to the treatment of virus infections. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00193-06 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one Una between the borders.) Molecular Biology of Hereditary Eye Diseases PRINCIPAL INVESTIGATOR (List o^er professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: George Inana M.D.,Ph.D. Head, Section on LMOD.NEI Others: Carmelann Zintz Ph.D. Yoshihiro Hotta M.D. Caroljm Chambers Ph.D. Tetsuo Sasabe M.D., Ph.D. Molecular Pathology Staff FeUow Visiting Associate IRTAFeUow Visiting Associate LMOD, NEI LMOD, NEI LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Molecular Pathology INSTITUTE AND LOCATION NEL Nm, Bethesda. MD 20892 TOTAL MAN-YEARS: 5.0 PROFESSIONAL: 5.0 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D {a1) Minors G (a2) Interviews IS (b) Human tissues n(c) Neither SUMMA RY OF WORK (Use standard unreduced type. Do rmt exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00003-17 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Pharmacology of Ocular Complications PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, dtle, laboratory, and institute affiliation.) PI: Peter F. Kador Ph.D. Research Chemist LMOD,NEI Others: Julia Derr B.A. Biologist Kosei Karino M.D. Visiting Associate Tadashi Mizoguchi Ph.D. Visiting Scientist Yiikio Takahashi M.D. Visiting Associate Sanai Sato M.D. Visiting Scientist LMOD, NEI LMOD, NEI LMOD, NEI LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Molecular Pharmacology INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL ^MN-YEARS: 4.1 CHECKAPPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews PROFESSIONAL: 4.1 OTHER: 0.0 D (b) Human tissues 1X1(0) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The role of the enzymes aldose reductase and aldehyde reductase in the onset and progression of compUcations associated with diabetes are being investigated. In the eye these compHcations include retinopathy, cataract, keratopathy, changes in pupil ftinction, and iris and cihary process structure changes. Under development are methods of pharmacological control of these enzjmies to either delay or prevent the onset and progression of these complications. Events leading to the formation of several types of cataracts, as well as methods for controlling the onset of these cataracts through pharmacological intervention, are also being investigated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00011-16 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (SO characters or less. Title must fit on one line between the tiorders.) Pigrment Dispersion With and Without Glaucoma PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS.NEI Others: Lessie McCain R.N. Nurse Specialist OGCS, NEI COOPERATING UNITS (if any) LAB/BRANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEL Nm, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.2 PROFESSIONAL: 0.1 OTHER: 0.1 CHECK APPROPRIATE BOX{ES) Kl (a) Human subjects [X] (a1) Minors IS (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The purpose of this project is to determine the risks of patients with pigment dispersion syndrome to developing glaucoma. Comparisons of patients with and without glaucoma will be made on the basis of diagnostic tests, genetic screening, aqueous humor dynamics, and pupillary responses to Hght. The data acquired may enable a determination of the risk of patients with pigment dispersion syndrome to developing glaucoma, as well as adding to the imderstanding of the pathology of the disease. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00060-12 OGCS PERIOD COVERED October 1. 1989 to September 30. 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between the borders.) Visual Function and Ocular Pigmentation in Albinism PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS.NEI Others: Lessie McCain Rafael Caruso R.N. Nurse Specialist OGCS, NEI M.D. Visiting Scientist OGCS, NEI COOPERATING UNITS (if any) UB^RANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEI. Nm, Bethesda. MP 20892 TOTAL MAN-YEARS: 0.20 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 0.15 OTHER: 0.05 □ (c) Neither B (a) Human subjects □ (b) Human tissues [X] (a1) Minors □ (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Patients with hj^omelanotic disorders such as ocular albinism, oculocutaneous albinism, Chediak-Higashi disease, Hermansky-Pudlak syndrome, and iris transillumination defects are being recruited to determine visual fimction with these conditions and to evaluate the changes in visual fixnction over time. Family members are evaluated to attempt to determine factors which may identify the heterozygous state. PHS 6040 (Rev. 1/84) GP0914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00083-13 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (SO characters or less. We must fit on one line between the tmrders.) Gyrate Atrophy of the Choroid and Retina and Other Retinal Degenerations PRINCIPAL INVESTIGATOR (List other professional personnel bekm the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS,NEI Others: Michael B. Gorin M.D., Ph.D. Lessie McCain R.N. Rafael Caruso M.D. Doris Collie A.A. Medical Officer Nurse Specialist Visiting Scientist Ophthalmic Health Technician OGCS, NEI OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) The Howard Hughes Medical Institute, Laboratory and the Department of Pediatrics, The Johns Hopkins University, School of Medicine, Baltimore, MD (David L. Valle, M.D.). UBB RANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda. MD 20892 TOTAL MAN- YEARS: 1.1 PROFESSIONAL: 0.5 OTHER: 0.6 CHECK APPROPRIATE BOX(ES) E (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space pmvided.) Patients with gyrate atrophy of the choroid and retina are examined systematically to confirm the diagnosis. Skin fibroblasts of affected patients and family members are grown in tissue culture and assayed for ornithine aminotransferase activity. The resvilts are evaluated for correlation with the presence of homozygosity or heterozygosity for the disease trait. Each patient is given a trial of pyridoxine to see if serum concentration of ornithine can be reduced; if so, the patient is classified as a "responder" and treatment with pyridoxine is continued. Nonresponder and responder patients are then placed on a low-arginine, low-protein diet with supplemental amino acids and observed for arrest or improvement of the disease. If patients are not considered eligible for the diet, or if they appear unable to comply with the dietary regimen, they are followed to record the natural progression of the condition. Patients with other forms of retinal degeneration such as retinitis pigmentosa, fundus flavimacvdatus, juvenile retinoschi- sis, and Usher's syndrome, are also examined and their courses are compared with those of gyrate atrophy patients. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PRCXJECT NUMBER ZOl EY 00163-08 OGCS PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Tide must St on one line between the borders.) NIH Interinstitute Genetics Program: The Genetics Clinic PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS,NEI Others: Michael B. Gorin Lessie McCain M.D., Ph.D. R.N. Medical Officer Nurse Specialist OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) Interinstitute Medical Genetics Program, NIH LAa^ RANCH Ophthalmic Genetics and Chnical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 0.3 PROFESSIONAL: 0.1 OTHER: 0.2 CHECK APPROPRIATE BOX(ES) S (a) Human subjects 1^ (a1) Minors n (a2) Interviews D (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space pmvided.) The Interinstitute Genetics Program and the Genetics Clinic, supported by the NIH Clinical Center, offer a multidisciplinary approach to patients with genetic disease (ZOl CP 05139-05 CEB). Involved in the program are researchers fitsm all Institutes. Patients evaluated in the clinic represent a broad spectrum of genetic diseases. During the last year, approximately 200 individuals seen represented about 60 distinct disease categories. Due to the high frequency of ocvdar involvement in many of the cases, almost all the patients were evaluated by Ophthalmic Genetics and Clinical Services Branch staff or were discussed in consultation. The Clinic serves as a source of interesting case material concerning patients with inherited or developmental abnormalities of the visual system. In addition to the Genetics Chnic, patients are seen for genetic consultation at the Maryland School for the Blind. This experience has resulted the recruitment of patients into NEI intramural protocols. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00172-07 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Tide must fit on one line between the tiorders.) Age-Related Maciilar Degeneration PRINCIPAL INVESTIGATOR (List other professional personnel bebw the Prindpal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS, NEI Others: Monique S. Roy M.D. Visiting Scientist CB, NEI COOPERATING UNITS (if any) LAB/BRANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.05 PROFESSIONAL: 0.05 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) S (a) Human subjects D (a1) Minors D (a2) Interviews D (b) Human tissues n(c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00211-05 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. TiSe must lit on one line between the borders.) A Double-Masked Controlled Randomized Clinical Trial of Topical Cysteamine PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute afliliation.) PI: Muriel I. Kaiser-Kupfer M.D. Chief OGCS,NEI Others: Lessie McCain Manuel Datiles R.N. Clinical Technician OGCS,NEI M.D. Visiting Scientist OGCS, NEI COOPERATING UNITS (if any) Human Genetics Branch, National Institute of Child Health and Hviman Development (Wil- ham Gahl, M.D., Ph.D.) LAB/BRANCH Ophthalmic Genetics and Chnical Services Branch SECTION Section on Ophthalmic Genetics INSTITUTE AND LOCATION NEI. NIH. Bethesda. MD 20892 TOTAL MAN- YEARS: 0.50 PROFESSIONAL: 0.25 OTHER: 0.25 CHECK APPROPRIATE BOX(ES) ^ (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues n(c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Nephropathic cystinosis is an autosomal, recessively inherited storage disease in which nonprot- ein cystine accumiilates within ceUular lysosomes due to a defect in lysosomal cystine tremsport. Ocular manifestations include photophobia, crystal deposits in the cornea, conjunctiva, and iris, and depigmentation of the retina. Systemic complications include the Fanconi syndrome and renal failure. Nine years ago cysteamine, a free thiol that depletes cystine from cells, was introduced in the therapy of cystinotic patients. Althotigh patients had improved growth and stabiUzed renal function, there was no noticeable effect on the accumulation of corneal crystals. Recent studies showed that corneal cells in tissue culture are readUy depleted of cystine by the introduction of cysteamine, making feasible the use of topical ophthalmic cysteamine to circvunvent the humoral route. After appropriate animal studies to test for compUcations revealed none, we began a double-masked clinical trial to test the efficacy of topical cysteamine (0.1%) in humans. Fourteen patients of ages less than 3 years were enrolled and randomized to 0.1% cysteamine. Five patients showed a significant decrease in crystals in the cysteamine-treated eyes. In order to test the effects of increasing the concentration of cysteamine eye drops in humans, a toxicity study was performed in rabbits, showing no adverse reactions. The results permitted an increase in the concentration to 0.5% for human use, and aU patients receiving 0.1% cysteamine were switched to 0.5%. An additional 5 yoxmg patients showed a significant decrease in treated eyes. Thus, of 18 young patients, 10 successfully had the code broken; of the remaining 8, 2 died, 3 discontinued medication, and 3 are still in the trial Due to the success in the yovmger patients, this study was expanded to include older patients, 3-31 years of age. The findings have been most exciting: seven patients have shown a significant decrease in crystals in treated eyes as well £is improvements in comfort, i.e, relief of pain and photophobia. This study has resulted in significantly improved quality of life for the successfully treated patients. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBUC HEALT>H SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00084-12 OGCS PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the Ixjrders.) Anterior Chamber Anomalies Associated With Glaucoma or Octdar Hypertension PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: CarlKupfer M.D. Director NEI Others: Miuiel I. Kaiser-Kupfer M.D. Lessie McCain R.N. Manuel B. Datiles M.D. Head Nurse Specialist Visiting Scientist OGCS, NEI OGCS, NEI OGCS, NEI COOPERATING UNITS (if any) UBffi RANCH Ophthalmic Genetics and Clinical Services Branch SECTION Section on Cataract and Corneal Diseases INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 0.05 PROFESSIONAL: 0.05 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) S (a) Human subjects n (al) Minors n (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Recent embryological research has indicated the role of the neural crest in contributing to all connective tissues anterior to the lens epithelivmi. Therefore, the group of developmental anomalies of the anterior chamber with glaucoma or ocular h)T)ertension is being reviewed. PHS 6040 (Rev. 1/84) GP09U-91B DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00153-08 LSR PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 diaraaers or less. Title must fit on one line between the borders.) Visual Motion and the Stabilization of Gaze PRINCIPAL INVESTIGATOR (List ottier professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Frederick A. Miles D.Phil. Senior Research Physiologist LSR, NEI Others: Urs Schwarz M.D. Thomas S. CoUett Ph.D. Claudio Biisettini Ph.D. James R. Carl M.D. Visiting Associate Visiting Scientist Visiting Fellow Expert LSR, NEI LSR, NEI LSR, NEI LSR, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Section on Oculomotor Control INSTrnjTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 4.7 PROFESSIONAL; 4.1 OTHER: 0.6 CHECKAPPROPRIATE BOX(ES) B (a) Human subjects n (a1) Minors n (a2) Interviews D (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) We have done a series of studies concerned with the dependence of various oculomotor and perceptual phenomena on viewing distance. Our previous experiments on monkeys had shown that the visual and vestibular ocular responses to translational distiirbances of the scene and of the obser\'er are hnear functions of the inverse of the viewing distance. Such dependence on proximity is appropriate for the vestibiilar reflexes that are stimtdated by the linear acceleration of the head and must transform signals from cartesian to polar coordinates. We attributed the synergistic visual reflex sharing of this property to shared nexiral pathways secondary to shared function and frame of reference. We now report that smooth piirsuit tracking in monkeys also shows this dependence on proximity, though somewhat less vigoroiisly. We have begun similar studies in hiiman subjects and have some preliminary data. Hum an responses to translational distiirbances of the upper torso generated compensatory eye movements that were linear functions of the inverse of the viewing distance and that were virtually identical to those previously obtained in monkeys. However, in stark contrast to those of monkeys, human ocular following responses associated with translation of the visual scene showed no such dependence on proximity. We svispect that the insensitivity of hximan ocular following to viewing distance may result from the impoverished nature of the stim-ulxis situation. Other studies on the scaling of size and depth cues with viewing distance by human subjects indicate that angular size is an important parameter in the scaling of depth, especially at greater distances when oculomotor cues are less reliable. PHS 6040 (Rev. 1/84) GP0914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00196-07 LRCMB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (80 characters or less. Title must fit on one line between the borders.) Molecular Genetics of the Eye and Ocular Diseases PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: John M. Nickerson Ph.D. Biologist LRCMB, NEI Others: Diane Borst Steven Bernstein Jing-Sheng Si David Saperstein Fintan Steele Gerald J. Chader Ph.D. Staff FeUow LRCMB, NEI Ph.D., M.D. Senior Staff Fellow LRCMB, NEI M.D. Visiting Associate LRCMB, NEI M.D. Rxtramiiral NRSA Fellow LRCMB, NEI Ph.D. NRG Fellow LRCMB, NEI Ph.D. Chief LRCMB, NEI COOPERATING UNITS (if any) University of Maryland Medical School, Baltimore, MD (M. Rodrigues); Jules Stein Eye Institute, UCLA, Los Angeles, CA (D. Father, B. Bateman, J. Ngo-Jones, R. Sparkes); Departments of Pathology and Ophthalmology, University of Virginia, Charlottesville, VA (F. Gonzalez-Fernandez) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Gene Regulation INSTITUTE AND LOCATION NEL Nm, Bethesda, MD 20892 rOTAL MAN-YEARS: 3.8 PROFESSIONAL: 3.8 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) D (a) Human subjects n (al) Minors n (a2) Interviews (b) Human tissues a(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Interphotoreceptor retinoid-binding protein (IRBP) is the first example of an extraceUvdar matrix protein that plays a role in transporting, buffering, or mediating the actions of retinoids and fatty acids in the interphotoreceptor space. This laboratory has isolated and characterized recombinant DNA molecules necessary for the study of the structure and expression of IRBP. We determined the primary structure of the IRBP gene and its protein, prerequisites to thorough study of IRBP gene expression. The DNA clones are important substrates that provide the tools for studies of how IRBP is synthesized and functions. IRBP is a single pol5rpeptide that contains four 300 amino acid long repeats, with 30%-40% identity among the repeats. These sequences have been helpful in the analysis of the uveitogenic peptides in IRBP. The cell in the retina that contains IRBP mRNA is the photoreceptor. IRBP mRNA is large and usually gives only one band on a Northern blot. But two sizes of IRBP mRNA are present in rat and mouse. Having analyzed the IRBP gene in m a ny species, especially the human, we have determined that there is only one IRBP gene per haploid genome. The chromosomal location of the IRBP gene is 10q21.1 for human and 14 for mouse, chromosomal localizations that rule out IRBP as the defective gene in many inherited eye disorders. Moreover, we have demonstrated that IRBP cannot cause fundus albipunctatus, a rare autosomal recessive form of stationary night blindness with a defect in the vitamin A cycle. The IRBP gene structure, which is compact for the size of the protein, has only three introns. The remarkable qiaadruplication within the gene suggests an interesting evolution, possibly involving a processed gene intermediate and two unequal crossovers. We have begun to characterize the elements regulating IRBP gene expression, including both cis-elements (the DNA sequences) and irons-acting factors (DNA binding proteins). We find two homologous areas of sequence in the 5' flanking regions of the bovine and mouse IRBP genes, one from -1 to -350 and another at -1200 to -1410. At least two blocks of sequence (one in each homologous area) and at least one protein of 120,000 MW form DNA-protein complexes in this promoter by gel-shift assays, DNAse footprinting, and Southwestern blotting. A region including sequences 1.7 kb upstream from the start of transcription is important for maximum expression in transient assays. Shorter segments have lesser promoter activity. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00075-12 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 charaaers or less. Title must fit on one line between the borders.) Immune Functions in Ocular Diseases of Obscure Etiology PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Robert B. Nussenblatt M.D. Clinical Director NEI Others: Marc de Smet M.D. Benjamin Rubin M.D. Barry Grubbs B.Sc. RasKid Mahdi Scott Whitcup M.D. Jan Lopez M.D. Rubens Belfort M.D. Alan G. Palestine M.D. Visiting. Scientist Senior Staff Fellow Biologist Biologist Senior Staff Fellow Visiting Fellow Visiting Scientist SpeciaTVolunteer LI LI LI LI LI LI LI LI NEI NEI NEI NEI NEI NEI NEI NEI COOPERATING UNITS (if any) University of Kunmie, Kurume, Japan (Manabu Mochizuki, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 1.02 PROFESSIONAL: 1.00 OTHER: 0.02 CHECK APPROPRIATE BOX(ES) S (a) Human subjects D (a1) Minors D (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) In vitro cellular immvme functions and Isrmphocjrte subsets are being studied in a masked method in patients with ociilar toxoplasmosis, pars planitis, Behcet's disease, geographic cho- roiditis, and chorioretinitis of unknown origin. Crude ocular antigens, purified uveitogenic soluble antigen (S-antigen), interphotoreceptor-binding protein (IRBP) of the retina, and uveitogenic fractions of the retinal S-antigen are being used in a lymphocyte microculture technique to evaluate the presence of cellvdar immxine memory in ocular tissues. In addition, piirified antigens from the toxoplasmosis organism are being tested in this in vitro system. A subgroup of patients with posterior uveitis has been identified as having this immunologic memory. Ljonphocs^te subsets in the blood and in the eye are being defined in these patients by monoclonal antibodies, which may shed light on the basic mechanisms of uveitis and may be used as a guide for specific immunologic therapy. The serum from these patients is also being evaluated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00092-11 LI PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Title must tit on one line t>etween the borders.) HLA, ABO, and B-cell Alloaatigens and Ocular Inflammatory Disease PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, latxiratory, and institute affiliation.) PI: Robert B. Nussenblatt M.D. Clinical Director NEI Others: Charles Egwuagu Ph.D. Staff Fellow LI, NEI COOPERATING UNITS (if any) L'Hopital de la Pitie, Paris, France (Phuc Le Hoang, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.03 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 0.03 OTHER: 0.0 D (b) Human tissues El (a) Human subjects n (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. n(c) Neither PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00094-11 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TFTLE OF PROJECT {80 characters or less. TldB must fit on one line between the borders.) ImTmme Mechanisms in Experimental Autoimmtme Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute atfiliation.) PI: Robert B. Nussenblatt M.D. Clinical Director NEI Others: Yujiro Fiijino Stephan Thurau Rashid Mahdi M.D. M.D. Evelsni Beraud M.D. Benjamin Rubin M.D. Phuc Le Hoang M.D. Visiting Associate LI, NEI Special Volunteer LI, NEI Biologist LI, NEI Visiting Associate LI, NEI Senior StafFFeUow LI, NEI Visiting Scientist LI, NEI COOPERATING UNITS (if any) LAB«RANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH. Bethesda. MP 20892 TOTAL MAN-YEARS: 1.35 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 1.25 OTHER: 0.10 D (b) Human tissues D (a) Human subjects D (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed tiie space provided^) This project has been terminated. KI(c) Neitlier PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES ■ PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00115-12 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Tide must lit on one line between the borders.) Cyclosporine Therapy in Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below Oie Principal Investigator.) (Name, tide, laboratory, and institute alliliadon.) PI: Robert B. Nussenblatt M.D. Clinical Director NEI Others: Marc de Smet M.D. Benjamin Rubin M.D. Scott Whitcup M.D. Juan Lopez M.D. Chi-Chao Chan M.D. Senior Staff Fellow LI, NEI Senior Staff FeUow LI, NEI Senior Staff Fellow LI, NEI Visiting FeUow LI, NEI Medical Officer LI, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. Nm, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.68 PROFESSIONAL: 0.68 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) ^ (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed tfje space provided.) Cyclosporine, an endecapeptide fungal product with specific anti-T cell characteristics, will be administered to patients with sight-threatening ocular inflammatory disease of non-infectious origin who have failed on either corticosteroid or cytotoxic agent therapy. This procedure will test cyclosporine's efficacy in the treatment of uveitis. Within the context of ongoing studies, the combined use of cyclosporine A and ketaconazole will be evaluated: Selected patients whose uveitis is well controlled on cyclosporine for 1 year or more are undergoing kidney biopsies for evaluatation of the long-term effects of this agent. The use of specific thromboxane A2 antago- nists to inhibit the renal toxicity of cyclosporine is being considered. An ongoing phase I/II randomized trial using cyclosporine A and G nears completion. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1, 1989 to September 30. 1990 PROJECT NUMBER ZOl EY 00015-25 LRCMB TITLE OF PROJECT {80 characters or less. Title must lit on one line between the borders.) The Cell Biology of the Vertebrate Retina PRINCIPAL INVESTIGATOR (List other prolessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute atfiliation.) PI: PaulJ. O'Brien Ph.D. Head, Section on LRCMB, NEI CeU Biology Others: Sylvia B. Smith Ph.D. Staff Fellow LRCMB, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Cell Biology INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 0.6 PROFESSIONAL: 0.6 OTHER: 0.0 CHECKAPPROPRIATE BOX{ES) D (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues IS(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The posttranslational modifications of rhodopsin include acylation, glycosylation, and chro- mophore addition. All appear to take place in the rod inner segment. The resulting molecules exhibit a slightly higher molecular weight than the mature rhodopsin in the outer segment and thus can be distinguished. This higher molecular weight is attributable to the larger ohgosac- charide normally found on nascent glycoproteins. Ordinarily, the large oHgosaccharide is trimmed and modified as the glycoprotein passes through the Golgi complex. The glycoprotein acquires its mature molecular size by the time it reaches the ceU surface. Rhodopsin behaves somewhat differently in that it still exhibits a sHghtly larger molecular weight after it has been inserted into the plasma membrane of the rod outer segment. Several Lines of experimentation have shown that these molecules possess a galactose residue not present in rhodopsin molecvdes that have been further sequestered in isolated rod outer segment disc membranes. The experi- mental methods included sensitivity to galactose oxidase and P-galactosidase as well as affinity chromatography on galactose-specific lectin matrices. As the rhodopsin molecules migrated firom the plasma membrane to the disc membranes, they lost labeled galactose and lectin-binding. Separation of plasma membrane and disc membrane Abactions confirmed that galactose was trimmed from the oligosaccharide. This unusual sequence of events may be related to disc morphogenesis insofar as tunicamycin blockage of oHgosaccharide synthesis results in failure of the rhodopsin-containing membranes to form new discs. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00016-23 LRCMB PERIOD CX)VERED October 1. 1989 to September 30, 1990 TrUE OF PROJECT (80 characters or less. Tide must lit on one line between tfw borders.) The Biochemistry of Normal and Dystrophic Retinas PRINCIPAL INVESTIGATOR (List ottier professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: PaulJ. O'Brien Ph.D. Head, Section on LRCMB, NEI Cell Biology Others: Sylvia B. Smith Ph.D. Jun Li M.D. StaffFellow IRTA Fellow LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) School of Veterinary Medicine, University of Pennsylvania (G. Aguirre, D.V.M.); Cullen Eye Institute, Baylor College of Medicine (R.E. Anderson, Ph.D., M.D.) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Cell Biology INSTITUTE AND LOCATION NEI. NIH, Bethesda. MD 20892 TOTAL MAN-YEARS: 1.6 PROFESSIONAL: 1.6 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) IHuman tissues 13(0) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Previous studies on the metabolism of polyunsaturated fatty acids in the retinas of miniature poodles had suggested that the essential fatty acid, linolenic add, having 18 carbon atoms and three double bonds, was not elongated and desaturated to docosahexaenoic add (DHA) with 22 carbons and six double bonds in the rod outer segments (ROS). This analysis was made 24 hovurs after intravitreal injection of labeled linolenic add. Further study revealed that some elongation and desaturation had taken place elsewhere in the retinas. A more detailed study ia rats showed that there was rapid conversion of linolenic add to several of the iatermediates in the pathway, but that the final desaturation step was extremely slow, as was the appearance of labeled DHA in ROS. Thus, the mammalian retina appears to be capable of converting small amoxmts of linolenic add to DHA but is probably dependent on extraocular sources for the DHA needed to replace large amounts of ROS disc membranes, which are imiquely enriched in this fatty add. Poodles affected with progressive rod-cone degeneration (prcd) have low blood levels of DHA, probably because of a defect in the final desaturating enzym.e. Several types of retinitis pigmentosa patients also exhibit low blood levels of DHA, which could resiilt in impaired ROS renewal. Attempts to identify the plasma proteins involved in transporting linolenic add and DHA have revealed that labeled linolenic add, administered by gavage, first enters the blood bovmd to very low density lipoprotein which transports it to the liver. After conversion to DHA, the labeled fatty add then appears to be transported by albumin. Identification of the carrier proteins will target candidate genes for studies of inherited retinal degenerations. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00256-02 LSR PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Infonnation Processing by Visual System Neurons PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Lance M. Optican Ph.D. Chief, NMS LSR,NEI Others: John W. McClurkin Ph.D. Arthur V. Hays B.A. BradJ. Zoltick M.A. Staff FeUow Electronics Engineer Computer Programmer LSR, NEI LSR, NEI LSR, NEI COOPERATING UNITS (if any) Laboratory of Neuropsychology, National Institute of Mental Health (Barry J. Richmond, M.D., Timothy J. Gawne, Ph.D., Emad N. Eskandar, B.A.) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Section on Neural ModeUng INSTITUTE AND LOCATION NEL Nm. Bethesda, MD 20892 TOTAL MAN- YEARS: 2.8 PROFESSIONAL: 1.9 OTHER: 0.9 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors □ (82) Interviews n (b) Human tissues ia(c) Neitlier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Visual perception depends on rich interactions among individual neurons. These interac- tions depend on mechanisms that encode, process, and transmit information among different visual areas of the brain. We are applying information theory to neurophysiological studies of behavior to learn what role neurons play in forming perceptions. We have already shown that neurons in different areas of the brain encode and transmit information about stationary, two-dimensional pictures that vary in form, brightness, and duration. In aU areas studied, neurons encode picture information iising a mviltidimensional temporal code. Neurons can transmit at least three times as much information using a multivariate temporal code as could be transmitted using a iinivariate strength code. We are now recording from individual neurons during visual discrimination tasks. It appears that neurons in the inferior temporal cortex send two types of messages during a pattern recognition task. One type of message describes the picture, whereas the other type of message indicates that a certain response is called for, without describing the eliciting picture. These results suggest that behavioral and physical parameters of a stimulus may be coupled within a temporal code. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00217-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must lit on one line between the borders.) Lymphocyte Migration in Experimental Autoimmune Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Alan G. Palestine M.D. Head, Section on LI,NEI Clinical Immxinology M.D. Clinical Director NEI M.D. Senior Staff FeUow LI, NEI Others: Robert B. Nussenblatt Jeffrey N. Bloom COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL hMN-YEARS: 0.26 PROFESSIONAL: 0.26 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors n (a2) Inten/iews D (b) Human tissues ISl{c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00219-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 characters or less. Title must fit on one line between the borders.) The Effect of Bromocriptine on Hmnan Uveitis PRINCIPAL INVESTIGATOR (Ust other professional personnel beiow the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Alan G. Palestine M.D. Head, Section on LI,NEI Clinical Immunology M.D. Clinical Director NEI M.D. Senior Staff Fellow LI, NEI M.D. Senior Staff Fellow LI, NEI M.D. Senior Staff FeUow LI, NEI Others: Robert B. Nussenblatt Janet L. Davis David C. Herman Jeffrey N. Bloom COOPERATING UNITS (if any) MetaboHsm Branch, National Cancer Institute (Marie C. Gelato, M.D.) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEL NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 0.91 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 0.91 OTHER: 0.0 n (b) Human tissues El (a) Human subjects n (a1) Minors D (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. n(c) Neitfier PHS 6040 {Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00220-04 U PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between the borders.) Endocrine Modulation of Inimiine-Mediated Eye Disease in Rats PR\NC\P^L\NWSJ\GAJOn(Ust other professional personnel below the PritKipallrivesdgator.) (Name, dde, laboratory, arid instituteaftiliatiori.) PI: Alan G. Palestine M.D. Head, Section on LI, NEI Clinical Immunology Others: Robert B. Nussenblatt David C. Herman M.D. Clinical Director M.D. StaffFeUow NEI LI, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEL NIH, Bethesda. MD 20892 TOTAL NMN- YEARS: 0.31 PROFESSIONAL: 0.31 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00221-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Intraocular Class II Antigen Expression in Endotoxin-Induced Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, tide, laboratory, and institute affiliation.) PI: Alan G. Palestine M.D. Head, Section on LI,NEI Clinical Immunology M.D. Clinical Director NEI M.D. Special Volimteer LI, NEI M.S. Biologist LI, NEI Others: Robert B. Nussenblatt Horst Helbig Rebecca Gurley COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEL NIH. Bethesda. MD 20892 TOTAL MAN-YEARS: 0.31 PROFESSIONAL: 0.31 OTHER: 0.0 CHECK APPROPRIATE BQX(ES) n (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00230-04 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or less. TMb must fit on one line between the borders.) Modulation of Retinal Vascular Permeability by Inflammatory Mediators PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, dde, laboratory, and institute affiliation.) PI: Alan G. Palestine M.D. Head, Section on LI,NEI Others: Rebecca Gurley M.S. Benjamin Rubin M.D. HorstHelbig M.D. Clinical Immunology Biologist Senior Staff Fellow Special Volunteer LI,NEI LI,NEI LI,NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN- YEARS: 0.43 PROFESSIONAL: 0.03 OTHER: 0.40 CHECK APPROPRIATE BOX(ES) D (a) Human subjects a (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00247-02 LI PERIOD COVERED October 1, 1989 to September 30. 1990 TFTLE OF PROJECT [80 charaaars or less. Title must tit on one line between the txtrders.) Autoiminunity to the Anterior Uvea in Patients with Uveitis PRINCIPAL INVESTIGATOR (Ust other professional personnei below the Principal Investigator.) (Name, title, laboratory, and institute alfiliadon.) PI: Alan G. Palestine M.D. Head, Section on Clinical Immunology LI, NEI Others: Rebecca Gurley M.S. Biologist LI, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Immunology SECTION Section on Clinical Immunology INSTITUTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 0.77 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 0.17 OTHER: 0.60 n (a) Human subjects H (b) Human tissues D (a1) Minors n (a2) Inten/iews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. n(c) Neither PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00243-04 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one ling between the borders.) Ocvilar Cells Cultured Under Normal and Diabetic Conditions PR\NCIPAI INWSVGAJOB (List other professional personnel below the Principal Invesigator.) (Name, title, laboratory, and institute affiliation.) PI: Bruce A. Pfeffer Ph.D. Senior Staff FeUow LMOD,NEI Others: W. Gerald Robison, Jr. Ph.D. Chief, Section on LMOD, NEI Pathophysiology COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Pathophysiology INSTITUTE AND LOCATION NEL NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 1.5 PROFESSIONAL: 1.0 OTHER: 0.5 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors D (a2) Interviews Kl (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Biosynthesis of extracellular matrix molecules was studied using cultured monkey retinal pigment epithelial (RPE) cells. In one set of experiments, native interphotoreceptor matrix (IPM) was labeled in vivo to parallel the apical expression of putative IPM components by cvdtured RPE. By polyacrylamide gel electrophoresis and subsequent autoradiography, we fovmd that cultured RPE produces several proteins whose electrophoretic mobiUty coincides with those of proteins in vivo. Therefore, cultured RPE may be a source of previously unidentified IPM constituents. Also shown was cultured RPE synthesis of two well-characterized basement membrane components, the attachment glycoprotein laminin and heparan sulfate proteoglycan (HSPG). While the depositon of the former was polarized to the basal side of the cells, the latter was released in insoluble form as part of the extracellular matrix substrate and secreted in soluble form into the culture media. Cultured RPE cells do not proteolytically "chip" the precursor to the core protein of HSPC, but instead release an intact proteoglycan with glycosaminoglycan chains attached to the fidl-size 400 kD protein moiety. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALT>1 SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT HHUJLCI NUMBfcH ZOl EY 00126-09 LMDB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (flO charaaers or less. Title must fit on one line between the borders.) Crystallia Genes: Structure, Organization, Expression, and Evolution PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Joram Piatigorsky Ph.D. Chief LMDB,NEI COOPERATING UNITS (if any) Section velo- UB/B RANCH Laboratory of Molecular and Developmental Biology SECTION Section on Molecular Genetics INSTITUTE AND LOCATION NEL NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 13.5 PROFESSIONAL: 13.5 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors n (a2) Interviews n (b) Human tissues IXl(c) Neither SUMMA RY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The structiire and expression of various crystallin genes have been investigated in humans, mice, chicken, squid, and jellyfish. The hiiman and mouse oB-, the chicken PA3/A1- and the major sqiiid crystallin genes have been fiilly sequenced; many other squid and one jellyfish crystallin genes have been cloned. Numerous cis-regulatory sites have been identified iu the 5' flanking regions of oA-, oB-, |3B1-, and squid crystallin genes, all of which include sequences known to bind transcription factors found in many tissues and involved in the regulation of other genes. In this connection, at least seven different cDNAs encoding zinc fingers have been cloned from the mouse lens. One of these cDNAs encoding a protein called cxA-CRYBPl is imphcated in the lens-specific expression of the mouse oA-crystallin gene and is similar to human MBP-1 and PRDII-BFl, which bind to the MHC class I and the p-interferon gene regulatory elements, respectively. The oA-CRYBPl binding site confers lens-preferred expression to the thymidine kinase promoter when tested in an SV40- transformed mouse lens epithelial ceU Line, but not in primary cultures of chicken lens cells. The chicken oA-crystaUin gene was foimd to use at least one and possibly two different sequences further upstream than the oA-CRYBPl site for its expresssion in the lens cells; one of these has a dyad of symmetry. The insert exon of the oA-crystallin gene was shown to be present among many mammals, indicating an early evolu- tionary appearance for this alternatively spliced coding sequence. The two 5-crystaUin genes were shown to be differentially expressed in lens, heart and brain in chicken embryos. The 51 gene is specialized for lens, while the 52 gene is preferentially expressed in non-lens tissues, consistent with 62 being the homologue of the argininosucdnate lyase gene. Finally, the chicken carbonic anhydrase II gene was shown to be regulated differently in lens and retina, and a functional promoter has been cloned. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00260-01 LRCMB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Molecular Biology of Outer Retina-Specific Proteins PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, titfe, laboratory, and institute affiliation.) PI: T. Michael Redmond PI1.D. Research Biologist LRCMB, NEI Others: Christian P. Hamel M.D. Gerald J. Chader Ph.D. Visiting Associate Chief LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Gene Regulation INSTITUTE AND LOCATION NEI, NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 2.1 PROFESSIONAL: 2.1 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors D (a2) Interviews n (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Retinal pigment epithelial (RPE) cells and the photoreceptor (PR) ceUs are functionally and developmentally closely integrated. During development of the outer retina in rat, the determi- nant recognized by the RPE-specific monoclonal antibody RPE-9 is first expressed at post-natal day 3 whereas the PR outer segments (OS) appear at day 5. The OS first appears where RPE cells are already expressing their determinant. RPE-9 recognizes a 67-kDa protein specific to the RPE. This protein is found in mammalian and avian RPE. A membrane-associated protein, it is probably non-glycosylated. We have begun to screen a bovine RPE cDNA Library for a cDNA for this protein. We have subcloned DNA fragments corresponding to the first two repeats of bovine IRBP into a bacterial expression vector. IRBP is involved in the transport of retinoids, a functional relationship between the RPE and the PR. The resultant expressed protein firagments will be tested for their ligand-binding and immunological properties. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00228-04 LI PERIOD COVERED October 1, 1989 to September 30. 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Study of Ocular Glial Cell Involvement in Uveitis PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Francois Roberge M.D. Visiting Associate LI, NEI Others: Robert B. Nussenblatt Rachel Caspi M.D. Clinical Director NEI Ph.D. Visiting Associate LI, NEI COOPERATING UNITS (ilany) LAB/BRANCH Laboratory of Immunology SECTION Section on Immunoregulation INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 0.82 PROFESSIONAL: 0.82 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00045-12 LSR PERIOD COVERED October 1, 1989 to September 30, 1990 TtTLE OF PROJECT (80 characters or less. Title must lit on one line between the borders.) Visuomotor Properties of Neurons in the Thalamus PRINCIPAL INVESTIGATOR (List other protessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute atf illation.) PI: David Lee Robinson Ph.D. Section Chief LSR.NEI Others: CaroHne Kertzman Verity J. Brown Eric M. Bowman Edmond J. FitzGibbon James R. Carl Ph.D. IRTA LSR, NEI Ph.D. Visiting Fellow LSR, NEI Ph.D. NRSA Fellow LSR, NEI M.D. Senior Staff Fellow LSR, NEI M.D. Expert LSR, NEI COOPERATING UNITS (if any) Developmental Endocrinology Branch, National Institute of Child Health and Human Development (Rich- ard Sherins, M.D.); Experimental Therapeutics Branch, National Institute of Neurological Disorders and Stroke (Irene Litvan, M.D.); Department of Anatomy, Howard University (Robert J. Covne, Ph.D.) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Section on Visual Behavior INSTITUTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 2.0 PROFESSIONAL: 1.4 OTHER: 0.6 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) A visual stimulxis appearing in the periphery causes a shift of attention. This change is not localized to the site of the visual stimulus, but facilitates adjacent regions. Parietal cortex neurons are modulated during this attentional behavior: cells with visual receptive fields at the focus of attention are suppressed, a mechanism thxis leading to facilitation of the responses of cells not at the focus of attention. When animals respond to visual targets that alternate between locations in a predictable way, reaction times to expected targets are faster than to than imexpected targets. Parietal cells respond best to the unexpected stimulus. These observations are consistent with the parietal cortex's containing a signal to shift attention. We have tested a group of Parkinson's disease patients on complex reaction time tasks. Although the disease produces a general impairment on all tasks, treatment with L-dopa selectively improves perfor- mance on only one. Responses which are "compatible" with the visual stimulus signaling them are speeded with L-dopa; responses "arbitrarily" associated with the triggering stimulus are unaffected by this treatment. These studies suggest that the basal ganglia, using dopamine, function in the initiation of movement. We have discovered that when a brainstem region involved in the initiation of head movements is electrically stimulated, the animal makes a brisk head movement. The size and amplitude of this movement depends on the intensity of the current and the starting position of the head. Injecting chemical tracers, we discovered that areas sending information to this brainstem region include the deep layers of the superior colUculus, the primary motor cortex, and the premotor cortex. The axons of cells in these areas reach the cervical spinal cord as well as other premotor, brainstem sites. These studies help explain the organization of systems for initiating head movements. PHS 6040 (Rev. 1/84) GPO 914-91 8 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00149-17 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Ultras tructure and Fiinction of the Cells and Tissues of the Eye PRINCIPAL INVESTIGATOR (List other prolessional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: W. Gerald Robison, Jr. Ph.D. Head, Section on LMOD,NEI Pathophysiology M.D. Visiting Associate LMOD, NEI Ph.D. Senior Staff FeUow LMOD, NEI Others: Nora Laver Bruce A. Pfefifer COOPERATING UNITS (ilany) UB/BRANCH Laboratory of Mechanisms of Ociilar Diseases SECTION Section on Pathophysiology INSTITUTE AND LOCATION NEL Nm, Bethesda, MD 20892 TOTAL MAN-YEARS: 4.5 PROFESSIONAL: 2.0 OTHER: 2.5 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews Kl (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This laboratory has developed the first rehable and convenient animal model for the very distinct group of lesions characteristic of himian backgrovmd diabetic retinopathy. Taking advantage of the fact that aldose reductase has a higher affinity for galactose than for glucose, which results in more intracellidar polyol acciimulation in galactosemia, we fed rats galactose for extended periods in an effort to produce diabetic-like retinal lesions. Galactosemia indeed induced diabetic-like microangiopathies that were more advanced and more Hke human diabetic lesions than those which develop in long-term diabetic rats. The galactose-fed rat exhibits capillary basement membrane thickening within 28 weeks; selective perics^te loss, capillary dilation, endotheUal cell proUferation, some tortuosity of vessels, and some acellularity within 33 weeks; microanevirysms, occlusions, and shunts within 66 weeks; and extensive regions of varicose capillary meshwork by 98 weeks, making the galactosemic rat a good model for diabetes-induced retinal microangiopathies that take longer to develop. This rat should also serve as a model for other polyol-related complications of diabetes. The galactose-fed rat model has distinct advantages over genetic or chemically induced models of diabetes for intervention studies: It shows lesions sooner, and upon removal from the galactose diet, it returns to a normal physiological state within a few days. We plan to determine appropriate times for intervention using different aldose reductase inhibitors, and to attempt, by dietary manipiilation, to produce rat models that develop the diabetic-like retinal angiopathies sooner. Also, using cell culture, we will investigate possible mechanisms of endotheUal cell proliferation and subsequent pathol- ogies. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00162-07 CB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must tit on one line between the borders.) Vitreous Fluorophotometry PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Monique S.Roy M.D. Visiting Scientist CB, NEI COOPERATING UNITS (if any) Biomedical Engineering and Instrumentation Branch, Division of Research Services, NIH (Peter Bungay, Ph.D.) LAB/BRANCH Clinical Branch SECTION Section on Retinal and Vitreal Diseases INSTITUTE AND LOCATION NEI. NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.4 PROFESSIONAL: 0.4 OTHER: 0.0 CHECKAPPROPRIATE BOX(ES) E (a) Human subjects D (a1) Minors n (a2) Inten/iews D (b) Human tissues n{c) Neitlier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00198-06 CB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Jitie must tit on one line between the borders.) Sorbinil Retinopathy Trial PRINCIPAL INVESTIGATOR (Ust other professional personnel beiow the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Monique S. Roy M.D. Visiting Scientist CB,NEI Others: James R. Carl M.D. Senior StafT Fellow CB.NEI COOPERATING UNITS (If any) National Institute of Diabetes and Digestive and Kidney Diseases, NIH (R. Silverman) LAB/BRANCH Clinical Branch SECTION Section on Retinal and Vitreal Diseases INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL MAN-YEARS: 0.7 PROFESSIONAL: 0.7 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) K (a) Human subjects D (a1) Minors D (a2) Interviews n (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project has been terminated. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00263-01 LI PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (SO characters or less. Tide must tit on one line between the borders.) Comparison of Surgical Treatment in Uveitis Patients With Glaucoma PRINCIPAL INVESTIGATOR (List o&iar professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Benjamin I, Rubin M.D. Senior Fellow LI, NEI Others: Robert B. Nussenblatt M.D. Clinical Director Marc de Smet M.D. Visiting FeUow Medical Officer Chi-Chao Chan M.D. NEI LI, NEI LI, NEI COOPERATING UNITS (if any) LAB^RANCH Laboratory of Immunology SECTION Section of Clinical Immunology INSTITUTE AND LOCATION NEI. Nm, Bethesda, MP 20892 TOTAL MAN-YEARS: 0.2 PROFESSIONAL: 0.2 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) Kl (a) Human subjects B (a1) Minors n (a2) Interviews 1X1 (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Four patients have been entered into the study. Three patients were randomized to trabeculectomy with S-fluoroviracil (5-FU) and one to the Molteno glaucoma implant. Two patients were black; two patients, white. Two patients are women; two, men. Three of the patients have aphakic uveitis, and one patient is phakic with uveitis. Intraocular pressiure in all four patients has been maintained between 12 and 20 miUileters of merciiry. CompUcations occurring postsurgically have included serious choroidal detachments in the three aphakic patients. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00237-05 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. We must fit on one line between the txrders.) Characterization of the Lens PRINCIPAL INVESTIGATOR (List other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute afliliation.) PI: Paul Russell Ph.D. Research Chemist LMOD.NEI Others: Takahiko Yamada M.D. Visiting Associate LMOD, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Cataracts INSTITUTE AND LOCATION NEI. NIH. Bethesda, MP 20892 TOTAL MAN-YEARS: 1.5 PROFESSIONAL: 1.5 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects D (a1) Minors D (a2) Interviews Si (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The advent of transgenic animals has made possible study of the influence and regulation of various genes on the development of an organism. However, this technique has not generally been used to develop ceU lines for use in tissue cidture. Tissue culture of the lens epithelium has been a goal of lens researchers because it may afford an opportunity to develop in vitro systems to test the efficacy of anticataract agents, as well as to study some mechanisms of cataract formation. A recently obtained transgenic animal has the T-antigen from the SV40 virus linked to the oA-crystalUn promoter. Cells from the lens of this animal that prohferate in the tissue cvdture environment have been shown to produce all the a-crystallins. These cells also sjmthesize the enzjmie aldose reductase, which is induced under conditions of hyperosmolarity. The cell line has been used to study metabohc alterations that occur dming incubation with naphthalene metabolites, compoimds chosen because of naphthalene cataracts' similarity to subcapsular age-related cataracts in humans. The specific-activity of the enzyme DT-diaphorase was in- creased when cells were exposed to high levels of naphthalene metabolites. Extensive work with epithelial samples from hiunan donor eyes has shown little difference between epitheUum from various sites in the lens or between epithelivun from young or old adult epithelium. PHS 6040 (Rev. 1/84) QP0 914-91B DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00252-02 LMOD PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT [80 characters or less. Title must fit on one line between the borders.) Cataract in the Philly Mouse Strain PR\HC\PAL\N\/ESl\Q^TOf^ (List other professional personnel below the Principal Investigator.) (Narne, title, laboratory, and institute affiliation.) PI: Paul Russell Ph.D. Research Chemist LMOD.NEI Others: Carolyn Chambers Ph.D. Staff Fellow LMOD, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Cataracts INSTITUTE AND LOCATION NEL NIH. Bethesda, MP 20892 TOTAL MAN- YEARS: 1.5 PROFESSIONAL: 1.5 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects D (a1) Minors D (a2) Interviews n (b) Human tissues ia(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) The Philly mouse derived from the Swiss-Webster strain develops a cataract about 6 weeks after birth. The results of early studies have shown that in the lenses of these animals, the epithehal cells fail to undergo complete differentiation. Biochemically, a 27 kD protein appar- ently missing from the Philly lens was shown to be the (3B2-crystallin, which iu the normal lens is a heat-stable protein. Investigation of the Philly mouse revealed that mRNA with approxi- mately the same size as the normal (362 mRNA is present in the Philly lens. It was further shown that a protein present in the PhiUy lens is immunologically related to the pB2 protein in the normal lens. This protein shares the same amino terminal as the normal PB2 but lacks part of the carboxyl half of the protein. The altered protein is shghtly smaller and has a more acidic isoelectric point than the normal lens pB2-crystallin. cDNAs were cloned and sequenced for normal and Philly mouse pB2-crystallin. The normal mouse PB2 cDNA is 725 (bp) in length and has 618 bp of open reading frame. Deduced amino acid sequences suggest that the normal mouse pB2 lacks a phosphorylation site at the C -terminal that is foiind in other mammals. The Philly mouse has a deletion of 12 nucleotides in the area encoding its foiirth motif. The properties of the protein encoded with this deletion appear to be consistent with the earlier protein findings and may be responsible for the cataract formation. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00250-03 LRCMB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT (flO characters or less. Titia must tit on one line between the borders.) Molecular Biology of Experimental Autoimmxine Uveitis PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Toshimichi Shinohara PI1.D. Head, Section on LRCMB, NEI Others: Tohru Abe Kotaro Eto S.Sunil M.D., Ph.D. M.D., Ph.D. M.D. Molecular Biology Visiting Associate Visiting Associate Visiting Fellow LRCMB, NEI LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) National Research Council, Division of Chemistry, Canada (H. Henry Mantsch, Ph.D.); Wills Eye Hospital, Philadelphia, PA (L.A. Donoso, M.D., Ph.D.) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Molecular Biology INSTITl/TE AND LOCATION NEL Nm, Bethesda, MD 20892 TOTAL MAN-YEARS: 1.0 PROFESSIONAL: 1.0 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) ^ (a) Human subjects la (a1) Minors n (a2) Interviews D (b) Human tissues n(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) We have previously determined amino acid sequences of human, mouse, rat and bovine retinal S-antigen and rat pineal gland S-antigen. Immunogenic sites and four uveitopathogenic sites of S-antigen were also determined. Two of the immunogenic sequences were highly conserved among these species. There are many proteins which have a similar sequence with a uveitopathogenic site in the National Biomedical Research Fovmdation data base. We chemically synthesized many peptides, some of which induced experimental autoimmune uveitis (EAU) and experimental autoimmune pinealitis (EAP) in Lewis rats, including synthetic peptides from yeast (Saccharomyces cerevisiae) histone H3, Escherichia coli hypothetical protein, potato proteinase inhibitor, hepa- titis virus protein, Moloney mxirine sarcoma virus protein, and Moloney mvirine leukemia virus protein. In addition, native yeast histone H3 was also capable of inducing EAU. Interestingly, the animals that were administered the yeast histone orally suppressed the EAU and EAP induction by either yeast histone H3 peptide or a S-antigen peptide. Thus, the peptides that have moleciilar mimicry cross-induced the tolerance. These findings provide a basis for autoimmune inflammatory diseases of the eye in humans. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00132-09 LRCMB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 charaaers or less. Title must fit on one line between the borders.) Molec\ilar Biology of Phototransduction PRINCIPAL INVESTIGATOR (Ust other professional personnel below the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: ToshimichiShinohara Ph.D. Head, Section on LRCMB, NEI Others : Kunihiko Yamaki Thoru Abe Shuji Suzuki Takanobu Kikuchi M.D., Ph.D. M.D., Ph.D. M.D., Ph.D. Ph.D. Molecular Biology Visiting Associate Visiting Associate Visiting Associate Visiting Associate LRCMB, NEI LRCMB, NEI LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) Division of Cancer Research, Mount Sinai Hospital, Toronto Ontario, Canada (Martin Breitman, Ph.D.) LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Molectdar Biology INSTITUTE AND LOCATION NEI, NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 4.2 PROFESSIONAL: 4.2 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (al) Minors n (a2) Interviews n (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) We have characterized the S-antigen genes from human and mouse, a kinase cDNA from hu m a n ; phosducin cDNAs from cow, rat, and human; Shuzin cDNAs from human and cow and a Shuzin gene from human. The gene sequences of the human and mouse S-antigens were determined. The S-antigen genes were approximately 50 kbp in length, contained 16 exons and 15 introns, and were comprised of 97% intron and 3% exon. The 5'-flanking regions of the genes, approximately 1.5 kbp long, have no known regulatory elements for transcription such as TATA, GC, or CCAAT boxes. Interestingly, the 5' flanking regions of the hvmian and mouse genes expressed tissue specific promoter activity in both in vitro and in vivo transcription assays as well as in transgenic mice. Several cDNAs of the phosducin from human, rat, and cow were isolated, and we determined their DNA sequences. Each sequence contains a Ser73 for phosphorylation by A-kinase. Sequencing results show that the phosducin in the retina and pineal gland have the same sequences and the same phosphorylation sites. This suggests that the functional role of this protein is the same in the retina and pineal gland. The functional role of the retinal protein Shvizin is imknown. We isolated several cDNAs and sequenced each of these from human and cow. The entire gene sequence of human Shuzin was also determined. This gene is composed of two introns and three exons and it has a highly repetitive sequence in the 5'-noncoding region. We have constructed fusion genes containing a 5'-flanking S-antigen gene sequence upstream of the bacterial gene chloramphenicol acetyl transferase (CAT). A hybrid gene containing the 5'-flank- ing region of the moiise S-antigen gene and the CAT gene was microinjected into transgenic mice. Those mice expressed CAT activity in the retina and pineal gland, suggesting that the 1300 bp-long S-antigen promoter has a tissue specific enhancer and promoter. S-antigen cDNAs were subcloned into two expression vector systems. The expressed protein was purified by gel filtration, and crystallization of this protein is now in progress. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00070-13 LRCMB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the txjrders.) Vitamin A and Ocular Tissues PR\NC\P^L\N\/£SJ\QATOR (Ust other professional personnel below the PrindpaJ Investigator.) (Name, titlejaboratory, and institute affiliati^^ PI: Barbara Wiggert Ph.D. Head, Section LRCMB, NEI on Biodiemistry Others: Ling Lee M.S. Chemist Todd Dimcan M.S. Biologist Gerald J. Chader Ph.D. Chief LRCMB, NEI LRCMB, NEI LRCMB, NEI COOPERATING UNITS (if any) Boston Univ. School of Medicine (C. Cornwall, PhD., G. Jones. Ph.D.); The Johns Hopkins Univ., Baltimore (R. Adler. M.D.): Univ. of Lund, Lund, Sweden (T. van Veen, Ph.D.): Univ. of Illinois College of Medicine, Chicago (D. Pepperberg^h.D., H. Ripps, Ph.D. j; Univ. of Pennsylvania School of Veterinary Medicine, Philadelphia (G. Aguirre, LAB/BRANCH Laboratory of Retinal Cell and Molecular Biology SECTION Section on Biochemistry INSTITUTE AND LOCATION NEI. NIH. Bethesda, MD 20892 TOTAL MAN-YEARS: 2.6 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 1.1 OTHER: L5 1X1 (b) Human tissues n(c) Neither D (a) Human subjects D (a1) Minors n (a2) Interviews SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) In the intact eye of the developing rd/rd, +/+ mutant mouse, interphotoreceptor-binding protein (IRBP), which is normally located extraceUularly in the interphotoreceptor matrix, remains intracellular. However, when removed from the eye and placed in short-term organ cxilture, retinas from these mutant mice demonstrate the capacity to sjnithesize and secrete IRBP normally until postnatal day 11 - 12. There may then be regulatory factors which control IRBP secretion in the intact eye. In studies of the physiological role of IRBP in the normal retina iising the toad (Bufo marinus) eye-cup preparation, IRBP was shown to be capable of promoting regeneration of rhodopsin in bleached ROS. Serum albumin, however, did not promote regeneration. In addition, it was shown that 11 -cis retinal was removed from the RPE by IRBP but not by serum albumin. It appears, then, that IRBP plays an active role in the vision process. In initial studies on IRBP in mouse eyes inoculated with a mvirine coronavirus, there was a highly significant decrease in IRBP by day 3 following inoculation. In addition, IRBP was no longer restricted to its normal location in the interphotoreceptor matrix but had diffused in the direction of the vitreous, reaching as far as the inner nuclear layer. PHS 6040 (Rev. 1/84) GPOgU-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00255-02 LMDB PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Origins, Structures, and Functions of Crystallins PR\NC\PA\.\NVEST\QAJOR (List other professional personnel bebw the Principal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: Graeme J. Wistow Ph.D. Visiting Scientist LMDB,NEI Others: Robert Kim M.D. Staff FeUow Hyong Kim B.A. Guest Worker Doug Lee Ph.D. NRC FeUow Joram Piatigorsky Ph.D. Chief LMDB, NEI LMDB, NEI LMDB, NEI LMDB, NEI COOPERATING UNITS (if any) Department of Biochemistry, Johns Hopkins University (G. Hart, L. Roquemore); Depart- ment of Biochemistry, Baylor University (W. O'Brien); Jules Stein Eye Institute, UCLA (J. Horwitz) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section on Molecular Genetics INSTITUTE AND LOCATION NEL NIH, Bethesda, MD 20892 TOTAL NMN-YEARS: 2.9 PROFESSIONAL: 2.4 OTHER: 0.5 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) We have continued to characterize novel taxon-specific crystallins including ri-crystalKn/al- dehyde dehydrogenase in primitive placental mammals and |j.-crystallin in marsupials. Both of these proteins appear to bindNAD(P) cofactors. We have also discovered that the ocAins subunit of a-crystalhn has a much more ancient origin than was expected; it is expressed in marsupials as well as in placental mammals. In an attempt to explain the anomalous subunit size of PBl in birds, we have foimd that this major cj^oplasmic protein is specifically glycylated in normal bird lenses but not in mammals. At the same time, we noted that a minor fi-action of a-crystallin subunits in both birds and mammals is modified with an 0-linked GlcNAc moiety, something which may have fimctional significance for the a-crystallLn/small heat-shock protein superfam- ily. In the analysis of the gene for x-crystaUin/a-enolase we have foimd that the gene promoter is highly active in lens explants and in cultured Uver and skin cells. This suggests that the basis for any lens-preferred expression of this gene must lie in other cis elements or in posttranscriptio- nal events. Alternatively the promoter may be responding to the stressed condition of the cultured cells in these experiments. PHS 6040 (Rev. 1/84) GP09U-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOl EY 00109-10 LSR PERIOD COVERED October 1, 1989 to September 30, 1990 TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Visuomotor Processing in the Primate Brain PR\NC\PAL\ti^ESJ\GATOR {Ust other professional personnel below the Prindpal Investigator.) (Narne, tiltejaboralory, and insbtine affiliati^^ PI: Robert H. Wurtz Ph.D. Chief LSR,NEI Others: Dwayne S. G. Yamasaki Ph.D. Staff Fellow Charles J. Duffy M.D.,Ph.D. Staff Fellow David M. Waitzman M.D., Ph.D. Staff FeUow Terence P. Ma Ph.D. Guest Researcher Douglas Munoz Ph.D. Guest Researcher LSR,NEI LSR, NEI LSR, NEI LSR, NEI LSR, NEI COOPERATING UNITS (if any) LAB/BRANCH Laboratory of Sensorimotor Research SECTION Section on Visuomotor Integration INSTITUTE AND LOCATION NEL NIH, Bethesda, MP 20892 TOTAL MAN-YEARS: 4.9 PROFESSIONAL: 3.1 OTHER: 1.8 CHECK APPROPRIATE BOX(ES) D (a) Human subjects n (a1) Minors n (a2) Interviews D (b) Human tissues Kl(c) Neither SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Experiments have concentrated on the investigation of the smooth pursuit eye movement system, which keeps the fovea of the eye centered on a moving target, and of the rapid or saccadic eye movement system, which moves the fovea of the eye from one object of interest to another in the visual field. The pursuit system studies concentrated on the recovery of the abiUty to make pursuit eye movements following pimctate lesions in the middle temporal cerebral cortical area (MT) as a model for studying the recovery of function following cerebral cortical damage. We foxind that large lesions that ehminated multiple areas related to visual motion processing produced deficits in the generation of pursuit that did not recover fully in 7 months. The contribution of other cortical areas beyond these motion-related areas was foimd to be limited. A neuronal correlate of this recovery is expansion of receptive fields in areas of MT cortex adjacent to the lesion. This receptive field expansion could provide information about the region of the visual field previously served by the damaged cells. Studies of the saccadic system concentrated on neuronal activity in the superior colhculus. Altering saccadic eye movements by electrical stimulation of the colliculus further supports the model showing that superior colhciilus activity is part of a feedback control system in the brainstem that controls the amphtude and the velocity in saccadic eye movement. Cells in the rostral pole of the colliculus were found to be related to the fixation of gaze by the monkey, suggesting that these cells may be related to a neural system that controls when the monkey makes a saccadic eye movement. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES - PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PERIOD COVERED October 1. 1989 to September 30, 1990 PROJECT NUMBER ZOl EY 00238-05 LMDB TITLE OF PROJECT {80 characters or less. Title must fit on one line between the borders.) Proto-Oncogene Expression During Lens Differentiation and Development PR\NC\PAL\N\/ESJlG/KTOR (List other professional personnel below the Principal Investigator.) (Nartie, title, laboratory, and institute affiliation.) PI: Peggy Zelenka Ph.D. Others: Barbara Brewitt Ph.D. Anita Dash Jo Ann Rinaudo Ph.D. John TaUan Ph.D. Geneticist Staff Fellow Howard Hughes Scholar IRTA FeUow Staff Fellow LMDB,NEI LMDB.NEI LMDB, NEI LMDB, NEI LMDB, NEI COOPERATING UNITS (ilany) Department of Surgery, New Jersey Medical and Dental College (Thomas Lysz, Ph.D.) LAB/BRANCH Laboratory of Molecular and Developmental Biology SECTION Section on Cellvilar Differentiation INSTITUTE AND LOCATION NEI. NIH, Bethesda, MD 20892 TOTAL NMN-YEARS: 4.9 CHECK APPROPRIATE BOX(ES) PROFESSIONAL: 3.9 OTHER: 1.0 D (b) Human tissues ISI(c) Neither n (a) Human subjects D (a1) Minors n (a2) Interviews SUMMA RY OF WORK (Use standard unreduced type. Do not exceed the space provided.) This project investigates the expression of proto-oncogenes during the differentiation of embryonic lens epithelial cells to form lens fiber cells and seeks to determine the specific function of the corresponding gene products in the developing lens. The normal developmental profile of each of three nuclear proto-oncogenes, c-myc, c-fos, and c-jun, has been investigated in order to correlate expression of these genes with cell growth, differentiation, and expression of lens-spe- cific genes. Information concerning proto-oncogene action in the lens is provided by studies of the exact timing of proto-oncogene expression in relation to cell division, cell differentiation, and expression of specific genes. In particular, we have attempted to correlate expression of specific proto-oncogene mRNAs with expression of mRNAs for proteins such as heat-shock protein (HSP70) and calpactin, whose expression and/or activity has been shown to be regulated by specific proto-oncogenes in other cell types. Finally, the effects of growth factors on proto- oncogene expression are being studied to provide a basis for organ culture of lenses. PHS 6040 (Rev. 1/84) GPO 914-918 DEPARTMENT OF HEALTH AND HUMAN SERVICES • PUBUC HEALTH SERVICE NOTICE OF INTRAMURAL RESEARCH PROJECT PROJECT NUMBER ZOIEY 00105-11 LMOD PERIOD COVERED October 1. 1989 to September 30. 1990 TITLE OF PROJECT {80 charaaers or less. Title must lit on one line between the borders.) Structure and Composition of Lens Crystallins with Respect to Cataractogenesis PRINCIPAL INVESTIGATOR (List other professional personnel below the Prindpal Investigator.) (Name, title, laboratory, and institute affiliation.) PI: J. Samuel Zigler, Jr. Ph.D. Research Biologist LMOD, NEI Others: XinyuDu M.D. Visiting Fellow Guo-TongXu M.D. Special Vohinteer Visiting Fellow Vasantha Rao Ph.D. Padmini Rao Ph.D. Visiting Fellow Xiao-Ian Cui M.D. Special Volimteer LMOD, NEI LMOD, NEI LMOD, NEI LMOD, NEI LMOD, NEI COOPERATING UNITS (if any) Department of Ophthalmology^, University of Tennessee (H.M. Jemigan, Jr.); Oakland University, Rochester, MI (V.N. Reddy); Alcon Laboratories (M. Lou); National Cancer Institute, (M. Krishna and P. Riesz); Centre for Cellular and Molecular Biology, Hyderabad, India (D. Balasubramjinian). LAB/BRANCH Laboratory of Mechanisms of Ocular Diseases SECTION Section on Cataracts INSTITUTE AND LOCATION NEI. NIH. Bethesda. MD 20892 TOTAL MAN-YEARS: 2.7 PROFESSIONAL: 2.7 OTHER: 0.0 CHECK APPROPRIATE BOX(ES) n (a) Human subjects n (a1) Minors D (a2) Interviews 1X1 (b) Human tissues n{c) Neitiier SUMMARY OF WORK (Use standard unreduced type. Do not exceed the space provided.) Lens crystallins are structural proteins that comprise over 90% of the dry mass of the lens. In cataracts, the crystallins are found to be heavily modified, particularly via oxidation. This oxidative damage is thought to be a critical factor in the etiology of lens opacification. This laboratory is working toward elucidation of the actual functions of crystallins in the normal lens. We are studying how normal lens function is affected by modification of crystaUin structure (e.g., by oxidation) or by change in the composition of crystalUns through the loss by mutation of a particular crystallin. In the guinea pig model we are studjdng, a mutation in the ^-crystallin gene causes cataract. We have now shown that ^-crystaUin is an enzyme/crystaUin which specifically binds NADPH and which is capable of functioning catalytically while the mutant form of zeta is enzymatically inactive, ^-crystallin, which catalyzes the reduction of a class of qiiinones, appears to be distinct from previously described quinone reductases. This system provides a imique opportunity to investigate the effects on the lens of a defined structural change in a m^or lens protein. Use of this system also allows us to address fundamental questions concerning the function(s) of enzyme/crystallins. Two major potential mechanisms of oxidative damage to the lens are photochemical processes and Fenton reactions involving metal ions and hydrogen peroxide (H2O2). In collaboration with Dr. D. Balasubramanian, we have quantitated the capacities of varioxis photosensitizers present in the lens to produce singlet oxygen and superoxide anion. In addition, we have evaluated the effects of H2O2 and copper on lens crystallins. PHS 6040 (Rev. 1/84) GPO 914-918 v H i: > ■■^■- ""■" fm Library, iyitdjng 19 National instityfes §1 Mgiffii LIBRARY 'l\^ Amazing Res Amazing Research. Amazing Help. http://nihlibraiy.nlh.gov 10 Center Drive Bethesda,MD 20892-1 150 301-496-1080 ,pM«X3y "-Hrs^Al^"