Myxovirus (influenza virus) resistance A (MxA) is an interferon regulated protein responsible for a specific antiviral state against viral infection. Our lab has previously shown that MxA is up-regulated by androgens in the normal prostate epithelial cells; however, there is no known role for MxA in cancer. Meta-analysis of different expression databases (e.g. NCBI GEO and Oncomine) suggested a strong inverse association between MxA expression and prostate cancer. To confirm these studies, we performed immunohistochemistry on normal prostate and prostate cancer tissues. Our results revealed that MxA expression was indeed decreased in cancerous as compared to normal prostate, indicating that MxA could be transcriptionally down-regulated in cancer. Previous studies indicated that MxA down-regulation could be due to a specific polymorphism in the proximal MxA promoter at position -88. This single nucleotide polymorphism G>T (rs2071430) is involved in modifying the gene expression and interestingly, it harbors an interferon-stimulated response element (ISRE) that is required for expression in response to interferons. The "T" allele restores whereas the "G" allele attenuates ISRE binding, resulting in increased or decreased MxA expression, respectively. Based on these observations we hypothesized that decreased expression of MxA in prostate cancer could be due to the rs2071430 polymorphism. We investigated this polymorphism in genomic DNA from equal number of disease free and prostate cancer samples. The results provide evidence that the GG genotype (low promoter activity) is higher in PCa (72%) as compared to normal (58.6%). The TT genotype (high activity) was higher in normal (5.7%) compared to PCa (2.4%) p

Id proteins are members of basic helix-loop-helix family. However, Id proteins lack the basic binding domain, which prevents DNA binding, and thereby regulates transcription. There are four members in the Id protein family termed Idl-4. In prostate cancer the expression of Idl and Id3 is high, whereas member Id4 expression is low. Decreased expression of Id4 is due to promoter hypermethylation in prostate cancer as well as many other cancers. This observation led us to hypothesize that Id4 acts as a tumor suppressor in prostate cancer. Furthermore, evidence suggests ectopic Id4 expression in metastatic prostate cancer cell line DU145 induced cell cycle arrest, apoptosis, and senescence. In this study, we expanded on these earlier studies to demonstrate that gain of Id4 attenuates cancer phenotype whereas loss of Id4 promotes cancer phenotype in prostate cancer cell lines DU145 and LNCaP, respectively. Upon over-expression of Id4 in DU145 cells (DU145+W4), there was an increase in apoptosis, due to decreased mitochondrial membrane potential (MMP) and increased expression of pro-apoptotic markers (PUMA, BAX, and p21). Inversely, silencing of Id4 in LNCaP cells (LNCaP-Id4) led to decreased apoptosis due to an intact mitochondrial membrane and decrease in the expression of pro-apoptotic markers (PUMA, BAX, and p21). Since BAX, PUMA, and p21 are direct transcriptional targets of p53, these results therefore prompted us to investigate the effect of Id4 on expression and activity of p53. LNCaP cells express wild-type p53. DU145 cells harbor mutant p53 (P223L and V274F), which lies within the DNA binding domain and abrogates p53 transcriptional activity. DU145 cells also express high levels of p53, due extended half-life. Surprisingly, there was decreased expression of p53 in DU145+Id4 cells associated with nuclear localization indicating enhanced transcriptional activity. We investigated p53 DNA binding and transcriptional activity. We determined that mutant p53 in DU145+Id4 cells was transcriptionally active evident by increased luciferase activity and binding of p53 to the promoters of its targets. In LNCaP-Id4, p53 expression was decreased which resulted in decreased p53 transcriptional activity and decreased DNA binding ability. The data suggested that Id4 can restore mutant p53 activity, which is a significant observation. Our results also suggest that Id4 promotes the assembly of a macromolecular complex involving CBP/p300 that results in acetylation of p53 at K373, a critical post-translational modification required for its biological activity. Loss of Id4 in LNCaP cells also abrogated wild type p53 DNA binding and transcriptional activity with concomitant loss of CBP/p300 requirement and decreased acetylation. In conclusion, we demonstrated that loss of Id4 promotes cancer phenotype in LNCaP cells. We also demonstrated that the tumor suppressor activity of Id4 is in part through regulation of CBP/p300 dependent acetylation and function of p53.

Placental dysfunction is implicated in the pathogenesis of preeclampsia. Chemical signals between the placenta and maternal circulation are a suspect cause of endothelial dysfunction and maternal hypertension. This study examined select lipid mediators of inflammation produced by the placenta. Patients were recruited from Virginia Commonwealth University’s pregnancy clinics and placentas were collected at delivery. Forty-eight-hour explant cultures of villous placental tissue were used to model lipid production. Electrospray ionization mass spectrometry was used to quantify concentrations of free lipids in the culture media. Bicinchoninic acid assays were performed to quantify protein in each culture for normalization of lipid data. After analysis, it was found that severity of preeclampsia was correlated with a unique lipid profile. Pro-inflammatory hydroxyeicosatetraenoic acids and sphingolipids were elevated. Aspirin usage in patients who developed preeclampsia was found to attenuate accumulation of isoprostane oxidative stress markers and thromboxane production while preserving omega-3-fatty acid and increasing prostacyclin levels.

Bladder cancer is the one of the most common malignancies of the genitourinary system and the overwhelming majority of those cases, approximately 90% in the United States(1), are of the urothelial/transitional cell histologic type. Small cell histologic type of bladder cancer is extremely rare with a mean frequency of 0.7% (1), and due to its rarity, there have not been any large phase III clinical trials in order to establish a definitive treatment regimen. We report here one such case of this rare type of bladder cancer and our approach towards treatment. A 69-year-old man had an incidental finding of microscopic hematuria during routine annual testing performed by his primary care physician. He was referred to a urologist for further evaluation, and in the interim, he began to experience symptoms of nocturia, dysuria and gross hematuria. Cystoscopy revealed a 5 cm sessile mass within the bladder and transurethral resection of the tumor was performed. Histopathological analysis of the tumor revealed muscle invasive poorly differentiated urothelial carcinoma with neuroendocrine features suggestive of small cell carcinoma. Follow-up systemic imaging only revealed multiple lesions in the liver, with the largest solitary liver lesion measuring 4.4 x 3.4 cm and no discrete lung lesions. Patient was started on palliative systemic chemotherapy with carboplatin and etoposide and follow-up imaging demonstrated excellent response after four cycles of treatment; however, follow-up imaging after the completion of 6 cycles of treatment demonstrated disease progression. Patient was referred for consideration of enrollment into any clinical trials; however, unfortunately no trials were found to be available. Patient was subsequently offered systemic treatment with single-agent immunotherapy with pembrolizumab. Due to development of left sided hydronephrosis, nephrostomy tube placement was performed and patient was also started on palliative radiation. Primary small cell carcinoma (SCC) of the bladder is an exceedingly rare malignancy and therefore, data is not readily available in order to guide treatment decisions. The most commonly administered regimen consists of etoposide with a platinum agent, and this regimen is extrapolated from the treatment of SCC of the lung. However, as for patients like ours, who had progression of disease in a short interval and are deemed primary treatment (platinum) refractory, the prognosis certainly becomes far more grim and the treatment choices even more limited. In sharing our treatment approach, we hope to be able to provide insight towards potential future treatment choices for this most-challenging diagnosis, primary small cell carcinoma of the bladder. (1) Blomjous CE, et. al. Small cell carcinoma of the urinary bladder. A clinicopathologic, morphometric, immunohistochemical, and ultrastructural study of 18 cases. Cancer. 1989 Sep 15; 64(6):1347-57.

Melanosis vesicae (or bladder melanosis) is a rare, benign condition referring to the presence of dark pigmented melanin deposits usually within the bladder mucosa. The clinical presentation can be mistaken for primary or metastatic melanoma of the bladder, and thus a histologic assessment is useful for ruling out a malignant diagnosis. All documented cases of melanosis vesicae have presented with urinary symptoms, including hematuria, symptoms of cystitis, incontinence and obstruction. This is a case report presenting a 57-year-old female with complaints of incomplete bladder emptying who met criteria for urinary retention. She underwent in-office cystoscopy and was found to have suspected melanosis vesicae. The diagnosis was later confirmed on pathology following a bladder biopsy. The next phase in care for this patient is planned sacral neuromodulation for treatment of urinary retention. Previous case reports of bladder melanosis have suggested an association with melanin deposition and inflammatory mechanisms and have not demonstrated malignant transformation during follow up. This is, to our knowledge, the first report of documented bladder melanosis in conjunction with urinary retention. Further studies are required to understand the etiology, clinical significance, and clinical correlation of melanosis vesicae with urinary dysfunction.

Prostate and breast cancer are the leading causes of cancer-related death in men and women, respectively, and metastasis is the primary factor underlying the high mortality rates.1 Snail transcription factor is an important molecule that drives prostate and breast cancer metastasis through the process of epithelial mesenchymal transition (EMT). Proteolytic enzymes that promote invasion and metastasis such as the lysosomal cysteine protease cathepsin L (Cat L) have been shown to degrade E-cadherin, promoting the epithelial mesenchymal transition (EMT).2 It has also been shown that silencing Cat L can inhibit transforming growth factor-beta (TGF-β)-mediated EMT by suppressing Snail transcription factor.3 Several recent studies have highlighted an additional unexpected localization and site of action for Cat L within the nucleus in breast, colon and prostate cancer.4 Natural products have been shown to be efficacious in prevention and possible treatment of cancer.5 Specifically, we have been studying Muscadine Grape Skin Extract (MSKE) as a possible candidate to inhibit Snail signaling. MSKE has previously been shown to promote prostate cancer apoptosis.6 We hypothesized that Snail promotes nuclear localization of Cat L, which promotes EMT associated with increased migration and invasion, and that antagonizing Snail-Cat L signaling would lead to mesenchymal epithelial transition (MET). We showed for the first time that MSKE promotes apoptosis through induction of endoplasmic reticulum stress response and autophagy. Additionally, MSKE could inhibit Snail-mediated EMT via scavenging reactive oxygen species. Moreover, Snail could promote nuclear localization of Cat L, which then promoted cleavage of CDP/Cux, increased Snail transcription and decreased E-cadherin transcription by direct promoter binding of cleaved CDP/Cux, leading to EMT associated with increased migration and invasion. Interestingly, Z-FY-CHO, a small molecule specific inhibitor of Cat L, as well as MSKE could antagonize this signaling by promoting nuclear to cytoplasmic re-localization of Cat L. Therefore, we have dissected novel mechanisms of action of Snail and how it can be antagonized by MSKE natural product.

Background Female stress urinary incontinence (SUI) greatly affects quality of life. The midurethal sling (MUS) procedure has been widely accepted as the standard of care treatment for SUI, although there is little information regarding patients' subjective reports of symptom improvement. Objectives The objective of this study was to identify clinical and demographic characteristics that predict subjective symptom improvement following MUS procedures in women with SUI. Materials and Methods The study design was retrospective cohort. Subjects included women who underwent MUS between 2006 and 2008, returned mailed surveys and met our predefined inclusion criteria. Pre-operative data included demographics, prior surgery, co-morbid diseases, urodynamics and concomitant reconstructive surgery. Subjective improvement was measured by score improvement on the UIQ-7, UDI-6, the UDI stress subscale and Question 3 of the UDI, "Do you experience urine leakage related to physical activity, coughing, or sneezing?" Results The mean age of the study sample was 57 years, parity was 2.5 and BMI was 28. Subjects with lower MUCP demonstrated more improvement on the UIQ-7. ΔUDI-6 stress subscale scores were more sensitive to symptom change than either the ΔUDI-6 or ΔUIQ-7. Older, menopausal subjects with urethral hypermobility and concomitant vaginal suspension showed less improvement than subjects without these characteristics. After controlling for urethral straining angle, PVR, menopause and time out from surgery, older age and concomitant vaginal suspension were associated with persistent post-op symptoms on the UDI-6 Question 3 and age remained the only variable associated with persistent symptoms on the UDI-6 stress subscale. Conclusion Concurrent vaginal suspension and advancing age were risk factors for persistent symptoms following MUS procedures in patients with SUI. Symptoms may recur after 24 post-operative months. Clinicians are encouraged to provide additional preoperative counseling to those women who are at greatest risk for persistent symptoms.

Objectives: Adherence to National Comprehensive Cancer Network (NCCN) guidelines for ovarian cancer treatment improves patient outcomes. The aim of this study was to assess disparities associated with ovarian cancer treatment in the state of Kentucky and central Appalachia. Methods: Data on patients diagnosed as having ovarian cancer from 2007 through 2011 were extracted from administrative claims-linked Kentucky Cancer Registry data. NCCN compliance was defined by stage, grade, surgical procedure, and chemotherapy. Selection criteria were reviewed carefully to ensure data quality and accuracy. Descriptive analysis, logistic regression, and Cox regression analyses were performed to examine factors associated with guidelines compliance and survival. Results: Most women were age 65 years or older (62.5%), had high grade (65.9%) and advanced stage (61.0%) ovarian cancer. Two-thirds of cases (65.9%) received NCCN-recommended treatment for ovarian cancer. The hazard ratio (HR) of death for women who did not receive NCCN-compliant care was 62% higher compared to the women who did receive NCCN compliant treatment (HR 1.62, 95% CI 1.11-2.35). Results from the logistic regression showed that NCCN-compliant treatment was more likely for: women age 65-74 years compared to age 20-49 (OR=3.32, 95% CI=1.32- 8.32), late stage compared to early stage cancers (OR 0.32, 95% CI 0.20-0.53), receipt of care at tertiary hospitals (OR=1.92, 95% CI=1.10-3.34), and privately insured compared to Medicaid (OR=0.31, 95% CI=0.13-0.77) or Medicare (OR=0.31, 95% CI=0.15-0.66). Conclusions: When the treatment of ovarian cancer did not follow NCCN-recommendations, patients had a significantly higher risk of death. Women were less likely to receive NCCN-compliant care if they were of younger age (20-49 years), had early stage disease, were not privately insured, or had care provided at a non-tertiary hospital.

Includes bibliographical references and index.

Thesis (Ph.D. in Nursing) -- University of Colorado, 2006. / Typescript. Includes bibliographical references (leaves 177-185). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;