Search Content

Matching Items (2)

Filtering by

Member of: Theses and Dissertations

Investigating the role of SGEF (ARHGEF26) in cell survival through its interactions with BRCA1 in TMZ-resistant Glioblastoma

Description

Glioblastoma (GB) is one of the deadliest cancers and the most common form of adult primary brain tumors. SGEF (ARHGEF26) has been previously shown to be overexpressed in GB tumors, play a role in cell invasion/migration, and increase temozolomide (TMZ) resistance.[3] It was hypothesized parental LN229 cell lines with SGEF knockdown (LN229-SGEFi) will show decreased metabolism in the MTS assay and decreased colony formation in a colony formation assay compared to parental LN229 cells after challenging the two cell lines with TMZ. For WB and co-immunoprecipitation (co-IP), parental LN229 cells with endogenous SGEF and BRCA were expected to interact and stain in the BRCA1:IP WB. LN229-SGEFi cells were expected to show very little SGEF precipitated due to shRNA targeted knockdown of SGEF. In conditions with mutations in the BRCA1 binding site (LN229-SGEFi + AdBRCAm/AdDM), SGEF expression was expected to decrease compared to parental LN229 or LN229-SGEFi cells reconstituted with WT SGEF (LN229-SGEFi + AdWT). LN229 infected with AdSGEF with a mutated nuclear localization signal (LN229-SGEFi + AdNLS12m) were expected to show BRCA and SGEF interaction since whole cell lysates were used for the co-IP. MTS data showed no significant differences in metabolism between the two cell lines at all three time points (3, 5, and 7 days). Western blot analysis was successful at imaging both SGEF and BRCA1 protein bands from whole cell lysate. The CFA showed no significant difference between cell lines after being challenged with 500uM TMZ. The co-IP immunoblot showed staining for BRCA1 and SGEF for all lysate samples, including unexpected lysates such as LN229-SGEFi, LN229-SGEFi + AdBRCAm, and LN229-SGEFi + AdDM. These results suggested either an indirect protein interaction between BRCA1 and SGEF, an additional BRCA binding site not included in the consensus, or possible detection of the translocated SGEF in knockdown cells lines since shRNA cannot enter the nucleus. Further optimization of CO-IP protocol, MTS assay, and CFA will be needed to characterize the SGEF/BRCA1 interaction and its role in cell survival.

ContributorsNabaty, Natalie Lana (Author) / Douglas, Lake (Thesis director) / Loftus, Joseph C. (Committee member) / School of Life Sciences (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)

Created2021-05

3D Robotic Assessment of Proprioception for Up, Down, and Back Directions

Description

Background. Proprioception plays a large role in everyday functioning, involving both information of body position and movement (Johnson & Panayotis, 2010). Clinical assessments of proprioception are largely subjective and are not reliable measures for testing proprioception in impaired or unimpaired individuals. Recent advancements in technology and robotics have brought about new assessments that involve position matching and other paradigms. However, the results are confined to the horizontal plane and only look at a very small subset of human proprioceptive ability. Objective. The present study looks to overcome these limitations and examine differences in proprioceptive sensitivity across different directions in 3D space. Methods. Participants were recruited from Arizona State University to perform a "same-different" discrimination test using a robotic arm. Each participant was tested along two of the three directions, and within each direction, proprioception at four distances (1-4 cm) was tested. Performance was quantified using percent correct, d' analysis, and permutation testing on median and variance values. Results. Proprioceptive sensitivity was significantly greater in the up direction vs. down and back across all distances. The greatest difference in sensitivity occurred at 3 cm; permutation tests using median and variance values from percent correct and d' found statistical significance at this distance in the up vs. down and up vs. back comparisons. Conclusions. There is evidence that proprioceptive sensitivity is greater in an anti-gravity direction (up), in comparison to gravity-assisted or gravity-neutral (down and back) directions.

ContributorsPatel, Megha (Author) / Buneo, Christopher (Thesis director) / Helms Tillery, Stephen (Committee member) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)

Created2016-12