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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.
Methods. Latinx parents and their 6th-8th grade children were recruited from eligible middle schools in Maricopa County to participate in a larger intervention study. A sample of parent-youth dyads from the first cohort of the larger study was selected for cross-sectional analysis of baseline data in this study (n=124). Participants completed a survey requesting demographics, family meal habits, and dietary intake. Participants were asked to report annual income, education level, and number of family meals in the home in the past week. They were also asked to complete an Acculturation Rating Scale for Mexican Americans, a 6-item Household Food Security Questionnaire, and a 26-question Dietary Screener Questionnaire. Analyses were run using Spearman’s Rank Correlation test and a Chi Square test of Independence.
Results. Mean daily youth intake of FV was 2.7 ± 1.4 cup equivalents, and daily youth intake of sugars from SSBs was 8.6 ± 4.9 teaspoon equivalents per day. Fifty percent of parents reported 7 or more family meals per week, while 38.7% reported 3-6 family meals per week and 11.3% reported 2 or fewer family meals per week. There was no significant association between family meal frequency and youth FV (r=-0.154; p=0.256) or added sugar from SSBs (r=0.027; p=0.807) intake. Similarly, results from Chi Square analyses suggested there was no association between family meal frequency and parent income level (p=0.392), Mexican-oriented acculturation level (p=0.591), Anglo-oriented acculturation level (p=0.052) and food insecurity (p=0.754). In contrast, a significant association between parent education and family meal frequency was found (p=0.014).
Conclusions. Parent education may play a role in shaping family meal practices in Latinx families. More research is needed to further understand this relationship and the relationship between family meal habits and youth dietary intake.