Objective: To evaluate the level of agreement between two self-reported medication adherence scales and prescription refill records in older adults. Design: Cross-sectional study Setting: Imperial Plaza; a retirement community located in Richmond, Virginia. Participants: 32 independent-living older adults, taking anti-hypertensive medications and filling their prescriptions at on-site Plaza Professional Pharmacy were recruited in the study. Methods: Participants’ 6 months refill records were obtained and Medication Possession Ration (MPR) was calculated. Participants were interviewed using Morisky Medication Adherence Scale (MMAS) & Brief Medication Questionnaire (BMQ). Kappa statistics was used to evaluate the level of agreement. Results: Poor level of agreement was found between refill records and MMAS (k=-0.004), refill records and BMQ belief screen (k=-0.09), regimen screen (k=-0.09), and recall screen (k =-0.004). Strong agreement was found between MMAS and BMQ regimen screen (k=0.79) and recall screen (k=0.87) respectively. Conclusion: Self-reported measure of adherence exhibited poor agreement with prescription refill records.

The Hsp90 chaperone complex associates with the telomerase enzyme, facilitating the assembly of the ribonucleoprotein complex. While previous data from our laboratory indicate that Hsp90 and p23 remain stably associated with (functionally active) telomerase, more recent experiments suggest that these chaperones associate with telomeres independent of telomerase, presumably through a specific interaction with telomere binding proteins. The current study examines the novel interactions between TRF2, TRF1, TIN2 and TPP1 and molecular chaperones (Hsp90, Hsp70, p23). In vitro and in cell experiments have shown an interaction between TRF1 and TRF2 and the molecular chaperones Hsp90 and Hsp70. Inhibition of Hsp90 using drugs that specifically block ATPase activity results in an increased association of TRF1 and TRF2 with Hsp90 to presumably stabilize the telomere associated proteins to the telomere. A definitive explanation as to the mechanisms underlying the chaperone/telomere associated protein interaction has yet to be determined and further studies examining chaperones’ contribution to telomere structure and function are underway. A better understanding of the telomeric proteins and Hsp90 and their roles in nuclear events is important, as both have extremely important functions in the cell. Our current working hypothesis is that chaperone proteins associate with TRF2, TRF1, TIN2 and TPP1 to facilitate telomeric protein-protein interactions and protein-telomere binding in both cancer and normal cells. The interaction between chaperones and telomere binding proteins may eventually provide a better understanding of telomeric structure and function. Defining the mechanisms of telomeric protein regulation is important in the development of new therapeutic approaches for targeting telomeres to induce dysfunction. Clinical trials are underway employing drugs targeting Hsp90 in cancer cells and given the results here, these Hsp90 compounds likely cause telomere alterations.

The formation of the myelin sheath is a crucial step during development because it enables fast and efficient propagation of signals within the limited space of the mammalian central nervous system. During the process of myelination, oligodendrocytes actively interact with the extracellular matrix (ECM). These interactions are considered crucial for proper and timely completion of the myelin sheath. However, the exact regulatory circuits involved in the signaling events that occur between the ECM and oligodendrocytes are currently not fully understood. Therefore, in this dissertation we investigated the role of a known integrator of cell-ECM signaling, namely, focal adhesion kinase (FAK), during oligodendrocyte morphological maturation in vivo and in vitro. Conditional and inducible FAK-knockout mice (Fakflox/flox: PLP/CreERT mice) were generated to observe the effect of FAK loss on myelination in vivo. The role of FAK during post-migratory premyelinating oligodendrocyte morphological maturation was explored in vitro using primary rat oligodendrocyte cultures in combination with siRNA or inhibitor treatment. When inducing FAK knockout just prior to and during active myelination in vivo, we observed a significant reduction in the number of myelinated fibers following initial myelination. In addition, our data revealed a decreased number of primary processes extending from oligodendrocyte cell bodies at this stage of development under induction of FAK knockout. In contrast, myelination appeared normal on postnatal day 28. Our in vitro data demonstrated that reduction of FAK expression or activity affected the oligodendrocyte process network maturation in a way that is unique to the ECM substrate present. More specifically, our data suggest that FAK restrains the expansion of the oligodendrocyte process network in the presence of fibronectin and promotes expansion of the oligodendrocyte process network in the presence of laminin-2. We showed that the FAK-mediated restriction of oligodendrocyte morphological maturation is tightly developmentally regulated; being highly active during early stages of post-migratory premyelinating oligodendrocyte morphological maturation but greatly diminished at the later stages of oligodendrocyte development. Thus, our data suggest that FAK controls the efficiency and timing of CNS myelination during its initial stages, at least in part, by regulating oligodendrocyte process network morphological maturation.

The collective microbial genomes within our gut(microbiome) represent a powerful metabolic force, leading many authors to call our GI flora an "organ within an organ", and the metagenomic sequencing of our microbiome, "the second human genome project". Bile acids, endogenously produced by the host liver, represent both a strong selective pressure for potential colonizers, aswell as substrates for microbial metabolism. Indeed, microbes have evolved enzymes to deconjugate bile salts, epimerize bile acid hydroxyl groups, and 7alpha-dehydroxylateprimary bile acids. The products of microbial 7alpha-dehydroxylation, secondary bile acids, are suggested by numerous lines of evidence to be involved in promoting colon carcinogenesis. 7alpha-dehydroxylating activity is a multi-step pathway, genes of which have only been identified in a small number of organisms within the genusClostridium. The biochemistry of this pathway has been largely worked out. The third step in the pathway is introduction of a delta-4-double bond; however, the gene product(s) responsible have not been identified. The baiCD and baiH genes were cloned, expressed and shown to have NAD-dependent 3-oxo-delta-4-steroid oxidoreductase activity showing stereospecificity for 7alpha-hydroxy and 7beta-hydroxy bile acid, respectively.In addition, bai genes were isolated from C.hylemonae TN271 by bidirectional genome-walking by PCR. This represents the first report of bai genes from a "low activity" 7alpha-dehydroxylating bacterium. The gene organization and sequence of the baiBCDEFGHI operon was highly conserved between C. hylemonae TN271 and the "high activity" 7alpha-dehydroxylating bacterium C. scindens VPI12708. The baiA gene was located by PCR using degenerate oligonucleotides. Bi-directional genome-walking revealed what appears to be several novel genes involved in bile acid metabolism which were also located in C. scindens VPI 12708. Expression of a 62 kDa flavoprotein and reactionwith [24-14C] 3-oxo-DCA and NADP resulted in a product of greater hydrophilicity than deoxycholic acid. The identity of this product was not determined. A second gene appears to share a common evolutionary origin with the baiF gene. A hypothesis is offered regarding the function of these homologues as Type III CoA transferasesrecognizing 5alpha-bile acids, or 5beta-bile acids (allo-bile acids). A third gene encodes a putative short chain reductase, similar in size and predicted function to the baiA gene, which may be involved in the final reductive step in the pathway. These novel genes also contained a conserved upstream regulatory region with the baioxidative genes. Finally, two genes were identified which may serve as potential drug targets to inhibit bile acid 7alpha-dehydroxylation. The first is an ABC transporter which may be co-transcribed with the other novel bile acid metabolizing genes, and what appears to be a bile acid sensor/regulator similar to the Tryptophan-rich sensory protein (TspO)/mitochondrial peripheral benzodiazepinereceptor (MBR) family of proteins.

State Medicaid programs were established to care for the poor by eliminating financial barriers and increasing their ability to be treated within the mainstream of the heath care system. The number of children eligible for Medicaid services is increasing, yet the number of Medicaid providers remains low. Health care providers cite failed appointments as being a major problem with Medicaid patients and one of the largest deterrents to participating. The purpose of this study was to determine whether a difference in appointment keeping behavior exists between Medicaid and non-Medicaid orthodontic patients. During a twelve-month period, a tally of appointments was kept for 707 active patients at Virginia Commonwealth University's Department of Orthodontics. Patients were categorized as either Medicaid or non-Medicaid and their appointment keeping behavior was evaluated. The results revealed that a significant difference does exist in the number of failed appointments between the groups (P<0.0001). The Medicaid patients failed 247(15.4%) of 1609 appointments and non-Medicaid patients failed 367(8.3%) of 4438 appointments. Additionally, these data show that although Medicaid patients accounted for only 26.6% of all appointments, they were responsible for about 40% of all appointment failures. The findings from this study support the concern among dental practitioners that Medicaid patients have higher rate of appointment failures than non-Medicaid patients.

The chemokine receptor CCR5 has been implicated in the pathogenesis of prostate cancer. A novel natural product, anibamine, was isolated and found to be a micromolar inhibitor of the receptor. Anibamine was used as a new anti-prostate cancer lead compound. To discover the pharmacophore, analogs of anibamine were designed using the “deconstruction-reconstruction-elaboration” approach and synthesized. The establishment of a stereoselective route to only one isomer was explored, to increase yield and eliminate elaborate purification procedures. Analogs were found to have anti-prostate cancer activity at levels higher than the parent compound. The molecular modeling studies of the deconstructed analogs indicate that due to the psuedo-symmetry of the parent compound, the binding conformation of the deconstructed analogs may not be very different from each other. All this information together may help identify a next generation lead compound for anti-prostate cancer treatment.

Worldwide, bacterial vaginosis (BV) is the most common vaginal disorder in women of childbearing age. BV is of major clinical importance due to its ability to significantly affect pregnancy outcome and enhance the transmission and acquisition of HIV. BV is characterized by a dramatic shift in the vaginal microflora; in most BV cases, the predominant bacterial species is Gardnerella vaginalis. It has been demonstrated that G. vaginalis forms an adherent biofilm on the vaginal epithelium of women with BV. Furthemore, evidence suggests that the high rate of recurrence associated with BV is related to incomplete eradication of the biofilm. The overall goal of this study was to characterize G. vaginalis virulence properties, including biofilm formation, in order to better understand the pathogenesis of BV and to improve available treatment methods. In an effort to tease apart the uncertain etiology of this disorder, we utilized in vitro assays to compare three virulence properties of G. vaginalis relative to other BV-associated anaerobes. Only G. vaginalis demonstrated all three virulence properties, including robust biofilm formation. It has been shown that the biofilm phenotype allows its constituent bacteria to be resistant to many negative environmental stimuli. Therefore, we studied the susceptibilities of biofilm vs. planktonic cultures to H2O2 and lactic acid. Biofilms tolerated higher concentrations of both chemicals; however, when the biofilm was proteolytically disrupted, sensitivity to the chemicals returned to planktonic levels. Since our data suggested a critical role for a protein in biofilm formation, a partial genome sequence of G. vaginalis was searched for sequence homology to known biofilm adhesins using the tBLASTn program. This revealed an open-reading frame encoding a hypothetical protein with significant homology to the staphylococcal Bap protein. Antibody towards a portion of the identified gene product was produced in rabbits by inoculation of a recombinant peptide to an antigenic region of the protein. Antibody inhibition assays against biofilm formation, adherence, initial adherence and aggregation were conducted. Relative expression levels of the biofilm-associated protein were analyzed under different conditions by western blot analysis. Finally, the protein was expressed in heterologous hosts and analyzed for an increase in biofilm formation.

The type II topoisomerase subfamily of enzymes has been clinically targeted by the widely used, broad-spectrum quinolone class of antibacterials. Due to emerging drug-resistant strains of bacteria, the quinolones’ effectiveness is threatened. The natural product simocyclinone D8 (SD8) has shown the ability to inhibit the type II topoisomerase, DNA gyrase, even when mutated to be resistant to the quinolones. In order to determine the pharmacophore required for SD8 binding to DNA gyrase, 16 compounds were synthesized. These compounds were then tested by surface plasmon resonance for their ability to inhibit the DNA – DNA gyrase binding interaction. It was found that three compounds were able to inhibit the DNA – DNA gyrase binding interaction, while another showed partial inhibition of the interaction. From this data, a minimum pharmacophore was able to be determined. The pharmacophore required a coumarin scaffold bonded to a carboxylic acid group through an approximately 15 Å hydrocarbon linker. Functional supercoiling assays determined that while the compounds were able to bind the enzyme, the binding did not inhibit DNA gyrase’s ability to supercoil DNA.

Background: Many factors affect whether women will insist that their sexual partners use a condom. This research project will identify some of the predictors of condom use among unmarried women in Ghana, Africa. Methods: This research project evaluated data from the 2008 DHS of women in Ghana, Africa for predictors of condom use among unmarried women. A total of 4,916 women completed the surveys. Of these, 1,966 women were unmarried. The research project employed multiple logistic regression analysis to determine condom use predictors at the time of the last sexual encounter for these 1,966 unmarried women. Results: Women with a secondary education or beyond were 3.2 (95% CI=2-5.2) times more likely to have insisted on the use of a condom than women with a primary education or less. Women ages 15-24 were 5.3 (95% CI=2.5-11.3) times more likely to have insisted on the use of a condom than women ages 35-49. Women ages 25-34 were not significantly different than the women ages 35-49. Women living in an urban area were 1.8 (95% CI=1.3-2.5) times more likely to have insisted on the use of a condom than women living in a rural area. Women with a higher level of literacy (could read a full sentence) were 3.6 (95% CI=2.5-5.1) times more likely to have insisted on the use of a condom than women with a lower level of literacy (were unable to read a full sentence). And in concert, women who read a newspaper or a magazine once a week or more were 2.4 (95% CI=1.6-3.5) times more likely to have insisted on the use of a condom than women who read a news paper or a magazine less than once a week. Similarly, women who watched television once a week or more were 2.9 (95%CI= 1.9-4.3) times more likely to have insisted on the use of a condom than women who watched less than once a week. Women who were determined to have “excellent” knowledge about HIV were 5.8 (95% CI=1.5-22.3) times more likely to have insisted on the use of a condom than women who were determined to have “fair or poor” knowledge. The results for women with a “good” knowledge were not significantly different than for those with “fair or poor” knowledge. Women who were determined, through a series of questions about wife beating, to have a low tolerance for abuse towards women (“strong” attitude about domestic violence/women’s rights) were 1.8 (95% CI=1.2-2.7) times more likely to have insisted on the use of a condom than women who were determined to have a high tolerance for abuse towards women (“poor” attitude about domestic violence/women’s rights). Results for women with a “fair” attitude were not significantly different from those with a “poor” attitude. Total life time sexual partners, frequency of listening to the radio, and interestingly, access to condoms did not significantly affect condom use. After multivariate adjustment, the significant predictors of condom use at the time of last sexual encounter were age, literacy, and amount of television watched. The results were: women age 15-24 (compared to women ages 35-49), women who could read a full sentence, and women who watched television once a week or more were 3.7 (95% CI=1.7-8.1), 2.1 (95% CI=1.4-3.3), and 1.8 (95% CI=1.2-2.8) times more likely to have insisted on the use of a condom during their last sexual encounter, respectively. Conclusion: Education, age, locality, literacy, media exposure (through reading the news paper or a magazine and watching television), knowledge about HIV, and attitude about domestic were predictors of condom use by the sexual partner of unmarried Ghanaian women at the time of last sexual encounter. After multivariate adjustment, only age, literacy, and amount of television watched were significant predictors of condom use at the time of the last sexual encounter.

Purpose: The purposes of this study were four-fold: 1) to determine the types and effectiveness of various topical anesthetics being used among dentists currently treating children, 2) to determine the types of procedures for which topical anesthetics are being used among children, 3) to understand the awareness and use of a relatively newer compounded topical gel Oraqix (Dentsply Caulk) among children, 4) to understand the adverse reactions to topical anesthesia that are seen among children. Methods: A cross sectional survey was designed, regarding the type, procedural use, effectiveness, and adverse reactions noted among children to various topical anesthetics. The survey sampled n=4933 actively practicing member dentists from a database of willing survey participants obtained from the American Academy of Pediatric Dentistry. The survey consisted of 14-items in multiple choice/answer format. The survey was pilot tested by a committee of faculty, and attached via e-mail with a cover letter containing a direct survey link for the study participants. Surveys were collected, posted, and managed through www.surveymonkey.com. Results: The study received 1255 responses from practitioners who are actively treating children giving an effective response rate of 25%. Of those that participated 94% are Pediatric dentists, 6% General dentists or “Other” specialists who treat children. The majority of respondents (95%) routinely use topical anesthetic, rating it as effective or very effective clinically. The most commonly used topical was 20%-Benzocaine gel with a reported 96% effective rate. The most common procedures topical anesthetics are being used for are pre-injection of local anesthetic and extraction of exfoliating deciduous teeth. Very few of the responding practitioners have ever heard of or used Oraqix gel prior to this survey. Many though, would consider using Oraqix if proven effective. Only 10% of respondents reported an adverse reaction to topical anesthetics, the most common being contact dermatitis or tissue sloughing from prolonged contact, followed by an allergic or aversive reaction to the dyes or flavoring in the topical anesthetic. Conclusions: The overwhelming majority of dentists treating children routinely use topical anesthetics to reduce pain response among children. 20%-Benzocaine gel is the most widely used topical anesthetic being used for dental procedures on children. Adverse reactions to topical anesthetic noted among practitioners treating children are very low but must still be strongly considered as potential life threatening risks if not used appropriately. Many practitioners treating children are still looking for the “ideal” topical anesthetic with improvements in taste, the ability to stay localized, the method of delivery, and improved effectiveness being key areas for future research.