Objectives

Accordingly, the specific aims of the BARB Core are:

• Provide services to DRC Regular and Supported members and Pilot and Feasibility Grant awardees in the specialized areas of mitochondrial bioanalytical analysis, oxidative stress assessment, and atherosclerotic lesion quantification using state-of-the-art research facilities, specialized expertise, and quality control
• Substantially reduce cost and investigator time commitment by accessing centralized resources, as opposed to operating independent laboratories and hiring separate faculty/staff
• Provide investigators and their technical personnel opportunities to learn and to further develop new research methodologies through recurring workshops and initial consultations to enable optimal usage of the various services of this core
• Pursue and develop methodologies that allow the investigator to extend capabilities and precision in redox biology and to foster multidisciplinary collaborations

Services

The BARB Core provides a series of molecular assays that will measure selected markers of oxidant stress, mitochondrial bioanalytical services, and quantification of atherosclerotic lesions. The selected methods are carefully geared to the needs of the DRC investigators, which is handled through a two-way process involving both the investigator and the core. 
The following sections briefly describe these methodologies.

E.1 Atherosclerotic Lesion Quantification
A basic service for aortic atherosclerotic lesion formation is provided by the BARB Core. Both the methods for cross-sectional and en face analyses have been published previously. This enables the association between the core's molecular measurements and resulting vascular pathology in diabetes and cardiometabolic disease.

E.2 Core Technologies and Functions
Space limitations preclude a detailed methodological description of all the services that will be offered by the core. The director and co-directors cumulatively have expertise in all the offered core services of oxidant stress markers, mitochondrial bioanalytical services, and atherosclerotic lesion quantification, as documented in the published references. 

Interaction with Other Cores
The BARB Core interacts with other DRC cores as well as non-DRC cores at UAB. The interactions with other DRC cores are critical to providing seamless services when assistance is required from multiple cores around a single project. Specifically, the BARB Core interacts with the Animal Physiology DRC Core for tissue collection; the Human Biology Core (e.g., plasma biomarker assays correlating with mtDNA damage and haplotype analyses), and Metrics (epidemiology of disease risk factors, mtDNA damage and haplotype assessment). Important interactions with cores outside the DRC include the UAB Comprehensive Cancer Center (CCC) Proteomics/Mass Spectrometry Facility (Dr. Stephen Barnes). The BARB Core provides a unique portfolio of services that does not duplicate any other UAB core facility.

Mitochondrial Bioanalytical Services

• mtDNA Damage Analysis
• High Resolution Respirometry
• Mitochondrial Oxidative Phosphorylation Complex and other Mitochondrial Proteins' Activity Assays
• Mitochondrial Complex I (NADH-Ubiquinone oxidoreductase) Assay
• Mitochondrial Complex IV (Cytochrome c oxidase) Assay
• Mitochondrial Citrate Synthase Assay
• Bioenergetics' Analysis of Tissues, Mitochondria, and Cells
• Mitochondria Isolation and Preparation

Aortic Lesion Assessment

• Aorta Lesion Assessment

Oxidative stress (Redox) Measures

• Glutathione and Ratios of Reduced (GSH) to Oxidized (GSSG)
• Aconitase Activity Superoxide Dismutase (SOD2; mitochondrial MnSOD) Specific Activity (expression and activity
• Oxidation of Thiols (reduced thiol levels measured)
• Lipid Hydroperoxide (LPO) Measures