Contribution of inflammation and growth factor expression to post-stroke cortical plasticity- Pilot
- UT Southwestern-Other
- Zale Lipshy University Hospital
Ann Stowe, Ph.D.
after collection, blood samples will be centrifuged to get pellets for Dna purification and genotyping. Genomic Dna will be extracted and primer pairs will be used to amplify BDnF polymorphism Val66Met, or VeGF polymorphism -2578C/a. Based on prior studies, we expect on the average of 41% of the initital enrollees to carry the BDnF Val66Met polymorphism and 66% to carry the VeGF-2578a/a polymorphism.
Following peripheral blood collection, each sample will be centrifuged to separate serum from cells. Serum will be used to quantitate chemokines, cytokines, and other growth factor levels, including VeGF and BDnF, using an enzyme-Linked immunosorbent assay (eLiSa) kit according to the manufacturer's instructions. Microplates will be coated with specific capture antibody, and serum samples aliquoted on the coated immunoassay plates. a corresponding biotinylated detection antibody will be applied and developed with avidin-horseradish peroxidase. a colorimetric substrate will allow for the intensity to convert absorbance to concentration for comparison of each biomarker.
The cellular portion of the blood will be stained for various cell-surface markers which will identify the composition of the cells in the blood, and allow the quantification of pro-inflammatory lymphocytes. an intracellular stain, carboxyfluorescein succinimidyl ester (CFSe), will be used to study the amount of auto-reactivity present.
We will retain whatever blood specimens that are left over after all the analyses are complete in case other tests become available that might be useful to explain inflammation and growth factor expression in post-stroke cortical plasticity.
in order to compare the degree of disability or dependence on others resulting from the stroke, research staff will assess all subjects who have experienced a stroke using the standard measure for neurologic disability, the modified Rankin Scale.
a maximum of 35 mls (2.5 tablespoons) of blood will be collected at each time point, for either the stroke patient, or the healthy control. The stroke patient's medical record will be accessed for information including, but not limited to, the patient's medical history, stroke history, and imaging results. The healthy control subjects will be self-reported healthy individuals. any medical history acquired from the healthy control subjects will be self-reported.
a total of 3 blood draws from stroke patients will be collected as follows:
acute Phase: 1 blood draw will be collected after consent, and within 0-48 hours after being
admitted to Zale Hospital
Sub-acute Phase: 1 blood draw between days 3 x 7 after hospital admission
Chronic Phase: 1 blood draw between 1 week and 3 months after initial hospitalization for
Stroke patients who are currently admitted at Zale Hospital will either have their blood drawn by the phlebotomists, Floor nurses at Zale, or by uTSW Phlebotomy Certified Research Staff. Stroke patients who are out of the hospital at the time we require a blood sample will be seen in the neuroscience Translational Research Center (nTRC) clinic in aston, where our uTSW Phlebotomy Certified Research Staff will draw their blood.
Healthy control subjects will have 1 blood draw at the time of consent. Healthy participants will have the option to be contacted in the future to provide additional blood specimens, as needed for research purposes. Sometimes, we find that a particular control subject may have a rare biomarker, or other test result, that provides a unique and important control for genetic polymorphism tests or inflammation tests.
Healthy control subjects will have their blood drawn at uT Southwestern or at the nTRC clinic in aston by uTSW Phlebotomy Certified Research Staff.
Inclusion Criteria for Healthy Control Subjects is as follows:
1.) Between 18-85 years old
2.) Self-reported Healthy volunteer
Inclusion Criteria for Stroke Patients is as follows:
1.) Between 18-85 years old
2.) Hospital admission due to an ischemic stroke, hemorrhagic stroke, or transient ischemic attack (TIA) x either a confirmed or suspected stroke or TIA diagnosis