Molecular Core – Summary The molecular and immunological characterization of inflammation and injury markers is crucial for the evaluation of the analgesic complementary approaches proposed in all four projects of this Center. Preliminary studies have identified sets of pro-inflammatory genes in sensory ganglia, spinal cord, brainstem and brain strongly induced by the pro-algesic stimuli in the models (chemotherapy, tibial fracture, functional pain syndrome, trigeminal pain) suggesting that pain has detrimental effects on the function of the CNS and induces inflammatory states in other organs remote from the site of injury. The Molecular Core will support the Center Projects through the following Specific Aims: Core Specific Aim 1: Analyze the effects of injuries, prior stress and complementary analgesic interventions on gene expression in injured tissues and neuronal tissues of the pain pathway. Core Specific Aim 2: Determine systemic and tissue levels of inflammatory biomarkers and inflammation-resolving lipid mediators. Core Specific Aim 3: Examine the effects of pain stimuli, prior stress and complementary interventions on systemic leukocyte profiles and neuroinflammatory cells in neuronal tissues. Core Specific Aim 4:Provide resources for production of viral tracers for labeling of neuronal circuits engaged in pain models and by bioelectronic complementary analgesic interventions. Methods include quantitative PCR analysis, multiplex immunoassays and ELISAs, flow cytometry and cell culture. The Molecular Core will work closely together with Project Investigators, Behavioral Core, the Statistical Unit of the Administrative Core and external core facilities to establish a workflow for sample tracing, blinded data transfer and feedback for optimization of experimental designs.