Breast Cancer Associated Gene 2 or BCA2 is identified as an E3 ligase in the ubiquitin proteasome system. It has been found that BCA2 is co-expressed with the estrogen receptor (ER) in ER-positive breast cancer cells. ER is crucial in the regulation of cell cycle progression and growth of breast cancer epithelial cells. Previous studies have shown that the BCA2 gene is regulated by the ER and BCA2 protein interacts with UBC9 and E2 ligase in the SUMOylation pathway. The regulation of BCA2 could prove critical since it could mediate the ubiquitination and possible SUMOylation of target proteins. While BCA2 as an Ubiquitin E3 ligase has been well established, its role in SUMOylation is unknown. The aim of this thesis is to identify BCA2’s involvement in SUMOylation. To achieve this, I performed bioinformatics analysis of the BCA2 protein sequence and found that out of the 6 lysines (Ks) in the sequence, K32 is the possible lysine important for SUMO attachment. I also tested the SUMOylation of BCA2 in an in vitro assay by using SUMO-specific proteins. The results have revealed an association between SUMO 2/3 and BCA2. Plasmids containing SUMO 1 and 2/3 were then purified from Top10 bacterial cultures and were transfected into HEK293 cells for immunoprecipitation experiments. Preliminary data from these experiments reveal a possible interaction of BCA2 with SUMO2/3. The involvement of BCA2 in both ubiquitination and SUMOylation pathways as an E3 ligase makes it a novel target for therapeutic intervention for the prevention and treatment of breast cancer patients.

Methods: DNA, RNA, and tissue from human kidney (n=77) were obtained. Kidney was chosen as target tissue because each of the four candidate genes and proteins (natriuretic peptide receptor 1 (NPR1), NPR2, NPR3 and membrane metallo-endopeptidase (MME)) are expressed there. DNA samples were genotyped for 120 single nucleotide polymorphisms (SNP) in these four genes using a custom Illumina array. mRNA levels were quantified using real-time reverse transcriptase polymerase chain reaction. Protein concentration was determined in tissue lysates using commercially available assays for each target. The association of SNPs with RNA and protein quantity was tested with ANOVA adjusted for gender and race. A principal components (PC) based method was also used to test the association of overall gene variation with RNA and protein quantity. Findings with false discovery rate (FDR) <0.05 were considered significant.

Results: Eleven SNPs in NPR2 were significantly associated with protein expression after controlling for FDR at 0.05, but none of these were associated with RNA quantity. Several genotypes in MME and NPR3 showed crude associations (unadjusted p<0.05) for protein and RNA quantity, but not after controlling for FDR. There were no SNPs associated with RNA or protein expression in NPR1. RNA and Protein quantity poorly correlated with each other; NPR1 and MME showed weak but significant positive correlations (p = 0.04 and 0.03, respectively). The PC analysis yielded similar overall results, with PC1 of NPR2 (accounting for 71% of variability) being the only significant association with protein (p=0.04), and no associations for RNA.

Conclusions: Genetic variation NPR2 is associated with altered protein expression, but this is not explained via gene expression. There were crude associations for MME and NPR3 variants but these did not reach adjusted significance in this small study. Additional studies are needed to assess the clinical impact, if any, of NPR2 genetic variation.

It begins with a discussion of the variety of obstacles, both private and public, that hindered women from entering the medical field. For some women, it was the responsibilities at home that left them little time and energy to pursue higher education. For others, it was simply a lack of access to the right opportunities and resources required to apply to medical school. During this time period, many medical schools and associations prohibited the acceptance of women. Certain homeopathic and sectarian schools were more accepting of women, although numbers were still limited.

It continues with a discussion of what the medical education, academic and social life, and opportunity were like for the women who did enter medical school. Here, I examine specific case studies of women who were students at this time. I include these women’s background information, their stories during medical school, as well as their career choices after graduation. I also discuss how the achievements of these women helped to gain increasing acceptance of women into medical school.

Finally, I conclude with a discuss of how these accomplishments helped shape the future of medical education and practice for women, as well as how and why women in medicine are still facing some of these challenges in their careers today.