Date of Submission

12-19-2023

Document Type

Thesis

Advisor

David San Pietro

Keywords

LDH, cardiac arrest, criminal investigations, hypoxia

LCSH

lactate dehydrogensase, cardiac arrest, criminal investigation, anoxemia

Abstract

In the realm of forensic casework, distinguishing between naturally occurring cardiac arrest deaths and those secondary to drug overdoses poses significant challenges. Pathologically, both scenarios culminate in respiratory failure, presenting difficulties in differentiation. A promising avenue for improvement lies in quantitatively measuring the duration of oxygen loss, or hypoxia, during cardiac events. Lactate dehydrogenase (LDH), an enzyme prevalent in the human body during anaerobic or hypoxic conditions, emerges as a candidate for this purpose. The expression ratio of LDH isoforms A and B, found in muscle and heart tissues, respectively, is hypothesized to help discern these cardiac events. Previous research conducted at the University of New Haven, titled “Development of qPCR-based LDH Detection Assay for Blood Samples as a Possible Marker for Hypoxia” and “LDH Isozymes as a Possible Marker to Differentiate Opioid Induced Hypoxia Death from Cardiac Arrest”, made sufficient progress toward quantifying LDH-A and LDH-B expression separately. This research aimed to compare their expression levels within the same blood sample using a larger cohort. Blood samples were obtained from 29 live donors aged 19-61, devoid of hypoxic cardiac events, to explore the consistency of LDH-A and LDH-B ratios in a healthy population. RNA was extracted from the whole blood samples, quantified, reverse transcribed, and the resulting gene expression was quantified with qPCR. Ultimately, the findings of this research were inconclusive. Future research should focus on optimizing cDNA concentrations for qPCR and employing an alternative positive control to EF1A. This preparation is crucial before transitioning to antemortem/postmortem blood samples from individuals who have undergone hypoxic cardiac events.

Available for download on Thursday, December 19, 2024

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