The objective of the proposed research is to yield information which will develop a clear understanding of human tear film stability in normal and diseased eyes, based on the roles taken on by various tear components. We propose to isolate and/or identify the responsible components. Special attention will be paid to the isolation of mucin. Other important tear components to be studied will also include plasma proteins, lipids, and enzymes. Microtechniques will be applied or developed. Methods of isolation and purification will include selective solvent extraction, gel and ion exchange chromatography, zone electrophoresis, gradient ultracentrifugation, immunological methods, and others. Microanalyses of individual components will provide a basis for the characterization and identification. Chemical analyses of isolates will include carbohydrates on GLC, amino acids and some amino sugars on the amino acid analyzer, and lipid components by TLC. Specific activities of some enzymes of importance will be assayed. Physical properties of individual components will include viscosity, solubility, molecular weight, surface activities and mode of binding of structural elements. Our preliminary studies on the subject: a) Fractionation and partial characterization of macromolecular components from human ocular mucus; b) O-Glycosidic linkage in the glycoprotein isolates from human ocular mucus, indicate that we will stand an excellent chance to achieve our objectives.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY004934-03
Application #
3259591
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1983-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
3
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Eye and Ear Hospital of Pittsburgh
Department
Type
DUNS #
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Chao, C C; Stuebben, A M; Butala, S M (1990) Characterization of ocular mucus extracts by crossed immunoelectrophoretic techniques. Invest Ophthalmol Vis Sci 31:1127-35
Chao, C C; Butala, S M; Herp, A (1988) Studies on the isolation and composition of human ocular mucin. Exp Eye Res 47:185-96
Chao, C C; Butala, S M; Jones, S L (1988) Association of serum albumin with tear prealbumin in human ocular mucus. Curr Eye Res 7:427-31
Chao, C W; Butala, S M; Zaidman, G et al. (1987) Immunological study of proteins and mucosubstance in saline soluble human ocular mucus. Invest Ophthalmol Vis Sci 28:546-54
Chao, C C; Butala, S M (1986) Isolation and preliminary characterization of tear prealbumin from human ocular mucus. Curr Eye Res 5:895-901