Blood banking, a meticulously regulated process endorsed by both the Food and Drug Administration (FDA) and the World Health Organization (WHO), plays a pivotal role in collecting and preserving red blood cells (RBCs) for transfusion medicine. Each day, the United States alone administers around 29,000 units of RBCs, addressing the diverse medical needs occurred from surgeries, diseases, traumas, and cancer treatments. However, conventional blood storage solutions employed, while serving as anticoagulants and preservatives, contain glucose levels that substantially exceed the typical "healthy" blood glucose range (4-6 mM). These solutions induce hyperglycemia, which is linked to significant negative alterations in RBCs. Vulnerable populations, such as people with diabetes, face severe consequences with chronic hyperglycemia. Transfusion medicine is undeniably lifesaving, but it is not without its drawbacks. It is theorized that adverse storage-related changes, referred to as storage lesions, are responsible for a multitude of post-transfusion complications. These storage lesion markers encompass metabolic and physical transformations that RBCs undergo during storage, with notable examples including advanced glycation end products (AGEs) and oxidative stress.High glucose concentration storage solutions adversely affect stored RBCs used for transfusion medicine. The Spence group proposed a potential alternative to the hyperglycemia issue: develop a normoglycemic blood storage solution. This novel RBC additive storage solution has already demonstrated improvements in key RBC function, such as adenosine triphosphate (ATP) levels and deformability. This dissertation reveals additional advantages through the evaluation of other storage lesion markers. Key components of this work include novel methods to detect and quantify AGEs and explore connections with oxidative stress, involving measuring free reduced glutathione (GSH).The core focus of this dissertation lies in investigating the ramifications of using a normoglycemic storage solution to reduce AGEs, particularly Nε-carboxymethyl-lysine (CML) and Nε-carboxyethyl-lysine (CEL), mitigate oxidative stress (as evidenced by GSH levels), and promote ongoing research involving normoglycemic storage conditions. This research is of paramount importance because it holds the potential to enhance the quality of RBCs for transfusion. Thus, directly impacting patient outcomes and quality of life, as well as offering insights into aging via in vivo patient samples. The methodologies employed in this study encompass the development of a pioneering approach utilizing ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) for the detection and quantification of CML, CEL, and lysine on the RBC membrane. A comparative analysis was performed between hyperglycemic (AS-1) and normoglycemic (AS-1N) storage solutions over 43 days. These findings played a critical role in determining AGE and GSH levels through weekly sample testing and involving innovative "feeding" techniques to ensure sterile normoglycemic glucose concentrations. In addition to comparing the two storage solutions, samples from type I diabetic (T1D) patients were utilized to explore the correlation between elevated blood glucose, glycated hemoglobin A1c (HbA1c%), and storage lesion markers. The methodologies and potential biomarkers presented in this study hold the promise of enhancing patient screenings and refining future C-peptide drug therapy clinical trials.In summation, this dissertation strives to pioneer novel methodologies and quantify storage lesion markers to advance transfusion medicine interventions. Ultimately, this research has the potential to improve the lives of countless individuals who depend on life-saving transfusions and future clinical trial drug discoveries.
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