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- Creators: Arizona State University
- Creators: Hale, Taben
- Member of: Theses and Dissertations
- Member of: Barrett, The Honors College Thesis/Creative Project Collection
Methods: Forty-two healthy six-week old male Sprague-Dawley rats were randomly assigned to one of three groups: plain water control, 0.6 mg/ml organometallic complex or 3.0 mg/ml organometallic complex as soon as they arrived. Each rat was then housed individually to prevent the sharing of microbiota through coprophagia. Rats in each treatment group were further divided into two dietary groups that were fed either a high fat diet containing 60% kcal fat that was changed every three days or standard rodent chow. Researchers were not blind to which rat was in each group. At the end of the 10-week study, rats were euthanized with an overdose of sodium pentobarbital (200 mg/kg, i.p.). Heart, left ventricle of the heart, liver, and spleen masses were recorded for each animal. Data were analyzed by two-way ANOVA using SigmaPlot 10.0 software.
Results: At the conclusion of this study, the left ventricle mass of the rats in the high fat diet group were significantly larger than those in the chow group. Neither dose of the organometallic complex supplement prevented these effects induced by high fat feeding.
Conclusion: The organometallic complex supplement was not effective at mitigating the effects of a high fat diet on cardiac hypertrophy in rats. Therefore, this supplement should not be used to treat cardiac hypertrophy.
Reducing the amount of error and introduced data variability increases the accuracy of Western blot results. In this study, different methods of normalization for loading differences and data alignment were explored with respect to their impact on Western blot results. GAPDH was compared to the LI-COR Revert total protein stain as a loading control. The impact of normalizing data to a control condition, which is commonly done to align Western blot data distributed over several immunoblots, was also investigated. Specifically, this study addressed whether normalization to a small subset of distinct controls on each immunoblot increases pooled data variability compared to a larger set of controls. Protein expression data for NOX-2 and SOD-2 from a study investigating the protective role of the bradykinin type 1 receptor in angiotensin-II induced left ventricle remodeling were used to address these questions but are also discussed in the context of the original study. The comparison of GAPDH and Revert total protein stain as a loading control was done by assessing their correlation and comparing how they affected protein expression results. Additionally, the impact of treatment on GAPDH was investigated. To assess how normalization to different combinations of controls influences data variability, protein data were normalized to the average of 5 controls, the average of 2 controls, or an average vehicle and the results by treatment were compared. The results of this study demonstrated that GAPDH expression is not affected by angiotensin-II or bradykinin type 1 receptor antagonist R-954 and is a less sensitive loading control compared to Revert total protein stain. Normalization to the average of 5 controls tended to reduce pooled data variability compared to 2 controls. Lastly, the results of this study provided preliminary evidence that R-954 does not alter the expression of NOX-2 or SOD-2 to an expression profile that would be expected to explain the protection it confers against Ang-II induced left ventricle remodeling.