Georgia Tech
1.
Orwig, Susan D.
Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease.
Degree: PhD, Chemistry and Biochemistry, 2011, Georgia Tech
URL: http://hdl.handle.net/1853/45816 
► The inherited form of primary open angle glaucoma, a disorder characterized by increased intraocular pressure and retina degeneration, is linked to mutations in the olfactomedin…
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▼ The inherited form of primary open angle glaucoma, a disorder characterized by increased intraocular
pressure and retina degeneration, is linked to mutations in the olfactomedin (OLF) domain of the myocilin gene. Disease-causing myocilin variants accumulate within trabecular meshwork cells instead of being secreted to the trabecular extracellular matrix thought to regulate aqueous humor flow and control intraocular
pressure. Like other diseases of protein misfolding, we hypothesize myocilin toxicity originates from defects in protein biophysical properties. In this thesis, the first preparative recombinant high-yield expression and purification system for the C-terminal OLF domain of myocilin (myoc-OLF) is described. To determine the relative stability of wild-type (WT) and mutant OLF domains, a fluorescence thermal stability assay was adapted to provide the first direct evidence that mutated OLF is folded but less thermally stable than WT. In addition, mutant myocilin can be stabilized by chemical chaperones. Together, this work provides the first quantitative demonstration of compromised stability among identified OLF variants and placing myocilin glaucoma in the context of other complex diseases of protein misfolding.
Subsequent investigations into the biophysical properties of WT myoc-OLF provide insight into its structure and function. In particular, myoc-OLF is stable in the presence of glycosaminoglycans (GAGs), as well as over a wide pH range in buffers with functional groups reminiscent of such GAGs. Myoc-OLF contains significant â-sheet and â-turn secondary structure as revealed by circular dichroism analysis. At neutral pH, thermal melts indicate a highly cooperative transition with a melting temperature of ~55°C. A compact core structural domain of OLF was identified by limited proteolysis and consists of approximately residues 238-461, which retains the single disulfide bond and is as stable as the full myoc-OLF construct. This construct also is capable of generating 3D crystals for structure determination. This data, presented in Chapter 3, inform new testable hypotheses for interactions with specific trabecular extracellular matrix components.
To gain further insight into the biological function of myoc-OLF, a facile fluorescence chemical stability assay was designed to identify possible ligands and drug candidates. In the assay described in Chapter 4, the target protein is initially destabilized with a chemical denaturant and is tested for re-stabilization upon the addition of small molecules. The assay requires no prior knowledge of the structure and/or function of the target protein, and it is amendable to high-throughput screening. Application of the assay using a library of 1,280 compounds revealed 14 possible ligands and drug candidates for myoc-OLF that may also generate insights into myoc-OLF function.
Due to the high â-sheet content of monomeric myoc-OLF and presence of an aggregated species upon myoc-OLF purification, the ability of myoc-OLF to form amyloid fibrils was suspected and…
Advisors/Committee Members: Dr. Raquel Lieberman (Committee Chair), Dr. A. (Yomi) Oyelere (Committee Member), Dr. Al Merril (Committee Member), Dr. Loren Williams (Committee Member), Dr. Nicholas Hud (Committee Member), Dr. Roger Wartell (Committee Member).
Subjects/Keywords: Open-angle glaucoma; Pharmacological chaperones; High-throughput drug screen; Amyloid fibrils; Gaucher disease; Myocilin; Glaucoma; Eye Diseases; Eye Diseases Genetic aspects; Intraocular pressure; Body fluids Pressure; Eye
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APA (6th Edition):
Orwig, S. D. (2011). Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/45816
Chicago Manual of Style (16th Edition):
Orwig, Susan D. “Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease.” 2011. Doctoral Dissertation, Georgia Tech. Accessed January 26, 2021.
http://hdl.handle.net/1853/45816.
MLA Handbook (7th Edition):
Orwig, Susan D. “Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease.” 2011. Web. 26 Jan 2021.
Vancouver:
Orwig SD. Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease. [Internet] [Doctoral dissertation]. Georgia Tech; 2011. [cited 2021 Jan 26].
Available from: http://hdl.handle.net/1853/45816.
Council of Science Editors:
Orwig SD. Biophysical and structural characterization of proteins implicated in glaucoma and Gaucher disease. [Doctoral Dissertation]. Georgia Tech; 2011. Available from: http://hdl.handle.net/1853/45816
2.
Binder, Nancy D.
Arterial blood pressure in the anephric sheep fetus.
Degree: PhD, 1980, Oregon Health Sciences University
URL: doi:10.6083/M4H70D0V
;
http://digitalcommons.ohsu.edu/etd/2318
Subjects/Keywords: Blood Pressure; Body Fluids; Extracellular Space; Fetus; Nephrectomy; Placenta; Renin
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APA (6th Edition):
Binder, N. D. (1980). Arterial blood pressure in the anephric sheep fetus. (Doctoral Dissertation). Oregon Health Sciences University. Retrieved from doi:10.6083/M4H70D0V ; http://digitalcommons.ohsu.edu/etd/2318
Chicago Manual of Style (16th Edition):
Binder, Nancy D. “Arterial blood pressure in the anephric sheep fetus.” 1980. Doctoral Dissertation, Oregon Health Sciences University. Accessed January 26, 2021.
doi:10.6083/M4H70D0V ; http://digitalcommons.ohsu.edu/etd/2318.
MLA Handbook (7th Edition):
Binder, Nancy D. “Arterial blood pressure in the anephric sheep fetus.” 1980. Web. 26 Jan 2021.
Vancouver:
Binder ND. Arterial blood pressure in the anephric sheep fetus. [Internet] [Doctoral dissertation]. Oregon Health Sciences University; 1980. [cited 2021 Jan 26].
Available from: doi:10.6083/M4H70D0V ; http://digitalcommons.ohsu.edu/etd/2318.
Council of Science Editors:
Binder ND. Arterial blood pressure in the anephric sheep fetus. [Doctoral Dissertation]. Oregon Health Sciences University; 1980. Available from: doi:10.6083/M4H70D0V ; http://digitalcommons.ohsu.edu/etd/2318
3.
Beitel, Adam Daniel 1993-.
THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER.
Degree: 2017, University of Saskatchewan
URL: http://hdl.handle.net/10388/8040
► The surface-mounted finite-height cylinder is a fundamental engineering shape and can be found in a multitude of industrial applications. As a result, the local flow…
(more)
▼ The surface-mounted finite-height cylinder is a fundamental engineering shape and can be found in a multitude of industrial applications. As a result, the local flow field is of great importance in the design of cylindrical components such as heat exchangers or buildings. While two-dimensional (2-D or “infinite”) cylinders are well-understood, the effects of the ground plane and the cylinder free end are significant and require further study. Of particular interest in this thesis is the
pressure distribution on the free end of the cylinder, and a mean normal force that develops from it. A vast majority of studies on this topic have focused on short cylinders with a small aspect ratio (AR = height/diameter). The work in this thesis is an attempt to characterize how the
pressure distribution and mean aerodynamic forces are influenced by the aspect ratio of the cylinder and the boundary layer thickness of the flow. The little-researched mean normal force, the mean drag force and its resultant mean bending moment, and the associated vortex shedding in the wake are investigated, along with the mean surface pressures and
pressure fluctuations for the cylinder free end. A cylinder was designed for use in measuring these parameters for 22 evenly spaced aspect ratios in a range from 0.5 ≤ AR ≤ 11, and an additional cylinder and boundary layer were used to generate data for four different values of relative boundary layer thickness in the range 0.60 ≤ δ/D ≤ 2.86.
The results of this research fit in well with published data, and reveal that the flow regimes appear to be marked by two critical aspect ratios, located approximately at AR = 2.5 and AR = 6. Below the lower critical AR, the boundary layer and ground plane effects are dominant, and the Strouhal number and the mean drag and mean normal force coefficients are drastically reduced. The mean bending moment coefficient is high at low AR, possibly owing to the high point of action of the drag force caused by the velocity distribution in the boundary layer. Between the two critical aspect ratios, the mean force coefficients and Strouhal number are relatively insensitive to AR. Above the upper critical AR, the mean drag coefficient increases towards the value for a 2-D cylinder, while the mean normal force coefficient reduces, and is expected to approach a small, constant value. A vertical wall shear force that acts in the opposite direction of the free end pressures may account for the difference between the mean normal force results obtained from integration and those obtained from direct measurements. For high AR, the bending moment coefficient and point of action are relatively unchanged. The free end
pressure distributions reveal similar features to previously published data, including “eye-like” enclosed regions of minimum
pressure on the upstream half of the cylinder face, and an enclosed region of maximum
pressure on the downstream half of the cylinder face. The eye-like structures disappear above the upper critical AR, and are replaced with a band of minimum…
Advisors/Committee Members: Sumner, David R, Bugg, James D, Mazurek, Kerry A, Evitts, Richard.
Subjects/Keywords: Fluids; bluff body; cylinder; drag; normal force; pressure distribution; free end
…x29;.
10
2.2 Wake System
The wake (a low pressure region behind the body) of a… …moment coefficient
CN
Mean normal force coefficient
CP
Mean static pressure coefficient
CP… …Static pressure RMS fluctuation coefficient
D
Cylinder diameter [m]
Ei
Load cell… …N·m]
n
Number of samples
P
Mean surface static pressure [Pa]
P0… …Stagnation pressure [Pa]
P∞
Freestream static pressure [Pa]
q∞
Freestream…
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APA ·
Chicago ·
MLA ·
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CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Beitel, A. D. 1. (2017). THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/8040
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Beitel, Adam Daniel 1993-. “THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER.” 2017. Thesis, University of Saskatchewan. Accessed January 26, 2021.
http://hdl.handle.net/10388/8040.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Beitel, Adam Daniel 1993-. “THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER.” 2017. Web. 26 Jan 2021.
Vancouver:
Beitel AD1. THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER. [Internet] [Thesis]. University of Saskatchewan; 2017. [cited 2021 Jan 26].
Available from: http://hdl.handle.net/10388/8040.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Beitel AD1. THE EFFECT OF ASPECT RATIO ON THE AERODYNAMIC FORCES AND FREE END PRESSURE DISTRIBUTION FOR A SURFACE-MOUNTED FINITE HEIGHT CYLINDER. [Thesis]. University of Saskatchewan; 2017. Available from: http://hdl.handle.net/10388/8040
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
4.
Cho, Eunhye.
Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes.
Degree: 2013, IUPUI
URL: http://hdl.handle.net/1805/3663
► Indiana University-Purdue University Indianapolis (IUPUI)
Articular cartilage is subjected to dynamic mechanical loading during normal daily activities. This complex mechanical loading, including cell deformation and…
(more)
▼ Indiana University-Purdue University Indianapolis (IUPUI)
Articular cartilage is subjected to dynamic mechanical loading during normal daily activities. This complex mechanical loading, including cell deformation and interstitial fluid flow, affects chondrocyte mechano-chemical signaling and subsequent cartilage homeostasis and remodeling. Focal adhesion kinase (FAK) and Src are known to be main mechanotransduction proteins, but little is known about the effect of mechanical loading on FAK and Src under its varying magnitudes and types. In this study, we addressed two questions using C28/I2 chondrocytes subjected to the different types and magnitudes of mechanical loading: Does a magnitude of the mechanical loading affect activities of FAK and Src? Does a type of the mechanical loading also affect their activities? Using fluorescence resonance energy transfer (FRET)-based FAK and Src biosensor in live C28/I2 chondrocytes, we monitored the effects of interstitial fluid flow and combined effects of cell deformation/interstitial fluid flow on FAK and Src activities. The results revealed that both FAK and Src activities in C28/I2 chondrocytes were dependent on the different magnitudes of the applied fluid flow. On the other hand, the type of mechanical loading differently affected FAK and Src activities. Although FAK and Src displayed similar activities in response to interstitial fluid flow only, simultaneous application of cell deformation and interstitial fluid flow induced differential FAK and Src activities possibly due to the additive effects of cell deformation and interstitial fluid flow on Src, but not on FAK. Collectively, the data suggest that the intensities and types of mechanical loading are critical in regulating FAK and Src activities in chondrocytes.
Advisors/Committee Members: Na, Sungsoo, Yokota, Hiroki, 1955-, Li, Jiliang.
Subjects/Keywords: FAK, Src, Mechanotransduction; Cells – Mechanical properties; Cells – Morphology; Cellular signal transduction; Tissue engineering; Cell adhesion molecules – Research – Methodology; Musculoskeletal system – Mechanical properties; Cell physiology; Osteoarthritis; Body fluids – Pressure; Tissues – Mechanical properties; Fluorescence spectroscopy; Articular cartilage – Pathophysiology
Record Details
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Record Details
Similar Records
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Cho, E. (2013). Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes. (Thesis). IUPUI. Retrieved from http://hdl.handle.net/1805/3663
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Cho, Eunhye. “Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes.” 2013. Thesis, IUPUI. Accessed January 26, 2021.
http://hdl.handle.net/1805/3663.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Cho, Eunhye. “Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes.” 2013. Web. 26 Jan 2021.
Vancouver:
Cho E. Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes. [Internet] [Thesis]. IUPUI; 2013. [cited 2021 Jan 26].
Available from: http://hdl.handle.net/1805/3663.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Cho E. Effects of interstitial fluid flow and cell compression in FAK and SRC activities in chondrocytes. [Thesis]. IUPUI; 2013. Available from: http://hdl.handle.net/1805/3663
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Open Access Theses and Dissertations
