Characterization of Glycation Sites on Human Serum Albumin using Mass Spectrometry

The modification of proteins by reducing sugars is a process that occurs naturally in the body. This process, which is known as glycation, has been linked to many of the chronic complications encountered during diabetes. Glycation has also been linked to changes in the binding of human serum albumin (HSA) to several drugs and small solutes in the body. While these effects are known, there is little information that explains why these changes in binding occur. The goal of this project was to obtain qualitative and quantitative information about glycation that occurs on HSA. The first section of this dissertation examined methods that could be used to quantify and identify glycation that occurs on HSA. The extent of glycation that occurred on HSA was quantified using oxygen-18 labeling mass spectrometry and the glycation sites were identified by observing the mass-to-charge (m/z) shifts that occurred in glycated HSA. This initial investigation revealed that oxygen-18 labeling based quantitation can be improved over previous methods if a relative comparison is done with oxygen-18 labeled peptides in a control HSA sample. Similarly, the process of making m/z shift-based assignments could be improved if only the peptides that were unique to the glycated HSA samples were used with internal calibration. These techniques were used in subsequent chapters for the assignment of early and late-stage glycation products on HSA. The regions of HSA that contained the highest amount of modification were identified, quantified, and ranked in order of their relative abundance. Of the commonly reported glycation sites, the N-terminus was found to have the highest extent of modification, followed by lysines 525, 199, and 439. The relative amount of modification on lysine 281, with respect to the aforementioned residues, varied with different degrees of glycation. The oxygen-18 labeling approach used for this analysis was novel because it allowed for the simultaneous quantification of all glycation-related modifications that were occurring on HSA. As such, several arginine residues were also found to have high amounts of modification on glycated HSA.

EFFECTS OF A BIRD HAZARD REDUCTION FORCE ON REDUCING BIRD/AIRCRAFT STRIKE HAZARDS AT THE ATLANTIC CITY INTERNATIONAL AIRPORT, NJ

Bird-aircraft strikes at the Atlantic City International Airport (ACY) increased from 18 in 1989 to 37 in 1990. The number of bird-aircraft strikes involving gulls (Larus spp.) during this time rose from 6 to 27, a 350% increase. The predominant species involved in bird strikes was the laughing gull (L. atricilla). Pursuant to an interagency agreement between the U.S. Department of Transportation (USDOT), Federal Aviation Administration (FAA) and the U.S. Department of Agriculture (USDA)l Animal and Plant Health Inspection Service (APHIS)/Animal Damage Control (ADC), ADC established a Emergency/Experimental Bird Hazard Reduction Force (BHFF) at ACY in 1991. An Environmental Assessment (EA) and Finding of No Significant Impact (FONSI) for the 1991 Emergency/Experimental BHRF was executed and signed by the FAA on 19 May 1991. The BHRF was adopted at this time by the FAA Technical Center as an annual program to reduce bird strikes at ACY. The BHRF goals are to minimize or eliminate the incidence of bird-aircraft strikes and runway closures due to increased bird activities. A BHRF team consisting of ADC personnel patrolled ACY for 95 days from 26 May until 28 August 1992, for a total of 2,949 person-hours. The BHRF used a combination of pyrotechnics, amplified gull distress tapes and live ammunition to harass gulls away from the airport from dawn to dusk. Gullaircraft strikes were reduced during BHRF operations in 1992 by 86% compared to gull strikes during summer months of 1990 when there was not a BHRF team. Runway closures due to bird activity decreased 100% compared to 1990 and 1991 closures. The BHRF should continue at ACY as long as birds are a threat to human safety and aircraft operations.