Master of Science / Biochemistry and Molecular Biophysics / Zhilong Yang / Poxvirus infections continue to threaten human health despite the eradication of smallpox, which was one of the most lethal infectious diseases in human history. Our objectives were to identify the host cell components/functions that are important for poxvirus infection and to gain insights into the molecular mechanism of poxvirus replication, ultimately guiding novel anti-viral development. Using vaccinia virus, the prototype poxvirus, we screened inhibitors of viral replication from over 3,000 chemical compounds, most of which have known cellular targets. This screening revealed numerous JAK/STAT3 inhibitors that could inhibit the replication of vaccinia virus. We further used multiple inhibitors of the JAK/STAT3 pathway and tested their effects on the replication of vaccinia virus in multiple primary and transformed cells through reporter assay and viral infectious particles measurement. The JAK/STAT3 inhibitors being tested were: SC144, an inhibitor of the interleukin 6(IL-6), a receptor of the JAK/STAT3 signaling pathway, AZ960 (a JAK2 inhibitor), Stattic and niclosamide (inhibitors of STAT3). Overall, our data indicate the JAK/STAT3 inhibitors could repressed vaccinia virus replication in multiple cell types, suggesting that the JAK/STAT3 signaling pathway is required for the efficient replication of vaccinia virus. Moreover, we observed that STAT3 was enriched in the cell nucleus, although the phosphorylation level of STAT3 was downregulated in vaccinia virus-infected cells during the early stages of infection. This study demonstrates an important role of the JAK/STAT3 signaling pathway in the replication of vaccinia virus, providing a possible novel direction by which to intervene in poxvirus infection and related diseases.

The feto-placental unit synthesises a variety of proteins and hormones which are secreted into the maternal circulation and amniotic fluid from early pregnancy. In pregnancies where the fetus has an autosomal trisomy, the normal concentration profiles of these markers in maternal serum and amniotic fluid are disturbed. These marker changes can be used to estimate the risk that a pregnancy is affected by Down's syndrome (or Trisomy 18) and thus allow the parents to make an informed decision regarding prenatal diagnosis by invasive testing. However, the factors which give rise to the varying patterns of marker concentrations in chromosomally abnormal pregnancies are poorly understood. The aim of this project was to investigate the underlying causes of abnormal marker concentrations in Down's syndrome, Trisomy 13, and Trisomy 18 pregnancies. The results of this investigation indicate that in Down's syndrome pregnancies, maternal serum levels of placental products reflect those found in the placenta; intact hCG, FβhCG and SP1 levels were elevated while PAPP-A and placental ALP levels were little changed. This suggests that transport of these proteins from the placenta into the maternal circulation is not affected but there is altered synthesis of hCG subunits and SP1. Hepatic synthesis of AFP does not appear to be altered in Down's syndrome pregnancies, but increased placental and reduced maternal serum levels of AFP point to a possible placental transport defect specific to AFP. Similarly reduced GGT levels in fetal intestine and in corresponding amniotic fluid from Down's syndrome pregnancies suggest that amniotic fluid GGT activity is of fetal intestinal origin since GGT activity was elevated in fetal liver and placental from the same series of Down's syndrome pregnancies.

Testicular self-examination (TSE) is an important behaviour to prevent the development of testicular cancer. This thesis examined the stages of decision making, emotions, perceived costs and benefits of screening, the self perceptions, social influences, and health routines in samples of adult males and adolescent school boys. A multi-method approach was adopted involving qualitative and quantitative methods of study. The qualitative part of the investigation comprised a series of studies including focus groups, in-depth interviews, programme evaluation and survey studies. The aim was to elucidate the meanings associated with health and illness and preventive cancer screening. Such information formed the bases for developing a survey measure and a health promotion programme to promote TSE. A comparative cross-sectional approach, including the components of three social cognition models was then carried out revealing a hybrid model (HSCM) to be most effective in explaining TSE. Acknowledging traditional social cognitions as predictors of TSE decision making, it also stresses the importance of emotional cognitions, self perceptions and routine behaviours. Using a five-stage model building approach, a longitudinal examination of stages of decision making was also carried out. Logistic analysis revealed that much of the variation in TSE practice was explained by a planning stage, implicating a two-stage motivation and volition stage rather than a five-stage model. In addition to explaining the characteristics associated with TSE practice and elucidating a stage approach to decision making, a draft health promotion programme was developed and evaluated. Two forms of the programme, a booklet and a seminar, were appraised and deemed effective as motivators of preventive action. Findings are discussed in terms of theory and practice of health education. The implications of the findings for health psychology are also discussed.

Class of 2012 Abstract / Specific Aims: The purpose of this study is to assess rural community members in Cochise County, Arizona, to determine if healthcare screenings and informational sessions are desired by the community members for chronic disease states, and from which healthcare providers the community members prefer to obtain these services. Identify which health professional rural members are most likely to seek for healthcare screening. Identify which health professional rural members are most likely to seek for healthcare informational sessions. Determine if rural community members are willing to pay for healthcare screening and informational sessions. Methods: This descriptive study obtained data through 350 questionnaires distributed during selected Cochise County community events in late 2011. Main Results: 341 surveys were completed. The majority of results favored physicians with 47.7% of the people stating they would very likely attend a health care screening conducted by a physician. 34.5% responded they would very likely attend an informational session conducted by a physician. Of the respondents willing to pay for health screenings, 63.9% would pay $30 or less. 70.9% stated they would not be willing to pay for an informational session. However, the majority of participants, 61.3%, indicated they would utilize pharmacists for OTC questions. Regarding questions about prescription medications, 43.4% would go to pharmacists, and similarly, 44% indicated they use physicians. Conclusions: As seen from the results, pharmacists are underutilized by rural community members; except for OTC questions. Respondents with higher levels of education were also more likely to prefer and attend events held by pharmacists.

Class of 2015 Abstract / Objectives: To develop an assay to permit the identification of compounds that can inhibit the interaction between GIT1 and the amino-terminal SH3 domain (SH3-N) of GRB2. Methods: The GIT1 protein was expressed in Sf9 insect cells and purified using Talon resin beads. The SH3-N domain of GRB2 was expressed in the E. coli strain, BL21(DE3)pLysS, and purified using glutathione resin beads. The SH3-N domain was fluorescently tagged on cysteine 32 using Cyanine 3 maleimide. The fluorescence of the assay was measured by using a plate reader with excitation wavelength of 555 nm and emission wavelength of 570 nm. Results: The GIT1 protein was expressed in Sf9 cells and purified using the Talon beads. The SH3-N domain of GRB2 was expressed in BL21 cells and purified from the glutathione resin beads. The SH3-N domain was cleaved from GST by using thrombin, which was engineered into the GST fusion protein and were fluorescently labeled using Cyanine 3 maleimide. Conclusions: The fluorescence polarization assay that will detect the interaction between GIT1 and the SH3-N domain of GRB2 is still under development, but it has progressed towards completion since both components of the assay are in hand.

M.Phil. (Biokinetics) / As with any other athletic activity, football participation carries an inherent risk for injury. Inadequate rehabilitation and incomplete healing have been identified as some of the key reasons for the high levels of injury in football. The majority of football injuries occur in the lower extremities. The aim of this study was to investigate the relationship between functional movement patterns and injury in junior football players. South African youth development players (119) participated in the study. This study adopted a quantitative and descriptive research approach. An injury questionnaire (retrospective) was completed and Functional Movement Screening (FMS) was used to assess each participant. The relationship between injuries suffered previously and FMS score was analysed. Kolmogorov-­‐Smirnov and Shapiro-­‐Wilk tests were used to determine normality. A Levene’s test for equality of variances was conducted. Independent sample t-tests were used to test inter-group differences. The confidence level was set at 95% (p≤0.05). The findings reflected that injuries, in this group, were prevalent with 87.3% of the sample having sustained a previous injury. Lower limb injuries were the most prevalent (76%); 58.8% of the lower limb injuries were of a non-contact nature and 41.1% were due to contact. The mean FMS score was 12.9, which, according to previous research, is linked to an increased risk for injury. There was a significant difference in FMS score between players who sustained a lower limb injury, compared to those who suffered an injury to another part of the body (p=0.032). This study found that injury was prevalent in junior football players and that FMS is a useful tool to screen for injury risk. The primary recommendation is for injury prevention strategies to be established in youth development football in South Africa.

Assisted reproductive technologies have become a common method used to treat infertility. These techniques have advanced quickly since the first birth of an in vitro fertilization (IVF) baby in 1978, at the Royal Oldham Hospital in the United Kingdom. Currently, IVF with or without intracytoplasmic sperm injection, is used throughout the world to achieve oocyte retrieval, fertilization, implantation of an embryo, clinical pregnancy, ongoing clinical pregnancy, and a live-born infant. The rationale for selecting one type of fertility treatment over another is multifactorial: the confirmed or unconfirmed cause of infertility, the age of the gamete donor and the recipient, the availability of the type of treatment, and the cost associated with the treatment. The ultimate goal of any fertility treatment is to achieve a successful pregnancy that results in a healthy infant. However, the literature is equivocal on the effects of fertility treatment cycles on the health outcomes of infants and mothers. Presently, there are thirty-six fertility treatment centres across Canada, eighteen of which reside in Ontario. A national, comprehensive database of assisted reproductive technology treatment cycles (Canadian Assisted Reproductive Technologies Register (CARTR) Plus) began collecting data in 2013, and has made the research objectives of this doctoral thesis feasible. Before this data collection system, population-wide studies involving fertility treatments were not possible in Canada. Two understudied issues associated with IVF are the impact of fertility treatments on the maternal serum screening markers used in prenatal screening programs to identify fetal aneuploidies; and the association between fertility treatments and adverse perinatal outcomes, such as preeclampsia and stillbirth. Given the increasing number of women who are using fertility treatments to conceive, it is imperative that studies investigating the association with adverse outcomes are conducted. As the science supporting fertility treatment procedure has advanced, so has prenatal screening. One of the first screening tests that are performed for newly pregnant women, including women who conceived following IVF, is maternal serum screening. The first objective of this doctoral thesis was to systematically review the literature on the association between IVF treatment and maternal serum screening marker levels and nuchal translucency (NT) thickness. After the search and screening of the literature there were 40 studies that were included in this systematic review. A decrease in pregnancy-associated plasma protein A (PAPP-A) and an increase in total human chorionic gonadotropin (hCG) was consistently reported for IVF pregnancies. However, since the levels of the other maternal serum screening markers reported also varied we were unable to generalize about the differences between prenatal screening results in the IVF population. These results led to investigating maternal serum screening marker levels among IVF patients in Ontario, Canada. The second objective of this thesis was three-fold: 1) to investigate the accuracy of IVF identification on the Ontario prenatal screening record, relative to reference standard on the CARTR Plus database; 2) to compare the prenatal screening markers in IVF versus non-IVF pregnancies in the population of Ontario; and 3) to propose updated IVF adjustment factors for prenatal screening in the Ontario population, based on the more accurate coding for IVF status in the CARTR Plus database. Significant differences between IVF and non-IVF groups, based on both the prenatal screening requisition information and CARTR Plus information, were found among the ethnicity adjusted mean multiple of the median (MoM)s for several prenatal screening markers: alpha-fetoprotein (AFP), PAPP-A, unconjugated estriol (µE3), first trimester hCG, total hCG, and dimeric inhibin A (DIA). When we developed the proposed adjustment factors for all CARTR Plus identified pregnancies we found that for PAPP-A, total hCG, and µE3 the mean adjusted marker MoMs were significantly closer to 1.00, as compared to the prenatal screening adjusted or the unadjusted mean marker MoMs. Currently, there is no adjustment made to the other maternal serum screening markers and NT measurement. The third objective was to examine the effect of type of infertility on placental-mediated adverse outcomes (preeclampsia, intrauterine growth restriction, placental abruption, and stillbirth). Type of infertility was classified as male factor (sperm count, poor sperm motility, and abnormal sperm morphology), female factor (ovulation disorders, tubal infertility, and uterine or cervical causes), and unexplained infertility. No significant associations were found between type of conception and the composite outcome, as well as each individual primary outcome. Similarly, the type of infertility was not associated with the composite outcome or any of the individual primary outcomes, except for female factor infertility, which was associated with increased probability of placental abruption. Overall, the results from this doctoral thesis suggest that there are substantial differences seen in maternal serum screening marker MoMs among women who use IVF to conceive, suggesting that appropriate adjustment factors should be employed to ensure accurate results for determining the risk of Down syndrome and trisomy 18. Additionally, although the literature has shown an association between fertility treatment and placental-mediated adverse outcomes no significant associations were found in the population of Ontario. Further studies should be performed to confirm the results of these observational studies.

Variable screening techniques are fast and crude techniques to scan high-dimensional data and conduct dimension reduction before a refined variable selection method is applied. Its marginal analysis feature makes the method computationally feasible for ultra-high dimensional problems. However, most existing screening methods for classification problems are designed only for binary classification problems. There is lack of a comprehensive study on variable screening for multi-class classification problems. This research aims to fill the gap by developing variable screening for multi-class problems, to meet the need of high dimensional classification. The work has useful applications in cancer study, medicine, engineering and biology. In this research, we propose and investigate new and effective screening methods for multi-class classification problems. We consider two types of screening methods. The first one conducts screening for multiple binary classification problems separately and then aggregates the selected variables. The second one conducts screening for multi-class classification problems directly. In particular, for each method we investigate important issues such as choices of classification algorithms, variable ranking, and model size determination. We implement various selection criteria and compare their performance. We conduct extensive simulation studies to evaluate and compare the proposed screening methods with existing ones, which show that the new methods are promising. Furthermore, we apply the proposed methods to four cancer studies. R code has been developed for each method.

Proper regulation of sleep and metabolism are critical to the survival of all organisms. In humans, dysregulation of sleep is linked to metabolic syndrome, including hypertension, hyperglycemia and hyperlipidemia. However, the mechanisms regulating interactions between sleep and metabolism are poorly understood. Although the fruit fly, Drosophila melanogaster, bears little anatomical resemblance to humans, it shares similar genetics essential in understanding normal development and disease in humans. From humans to flies, many disease-related genes and pathways are highly conserved, rendering the fruit fly ideal to understanding the interactions between sleep and metabolism. Therefore, using the fruit fly provides a framework for understanding how genes function between sleep and metabolism. During starvation, both humans and rats reduce their sleep. Similarly, previous studies have shown that fruit flies also suppress sleep to forage for food, further showing that sleep and metabolism are intricately tied to one another and that they are highly conserved across species. To further explore the interactions between sleep and metabolism, I have conducted multiple genetic screens to identify novel regulators of sleep-metabolism interactions. These experiments led to the identification of the mRNA binding protein translin (trsn) as being required for starvation-induced sleep suppression. A second screen that targeted metabolic genes from a genome-wide association study identified the ion channel accessory protein uncoordinated 79 (unc79) as a critical regulator of both sleep duration and starvation resistance. The genes function in different regions of the brain and suggest complex neural circuitry is likely to underlie regulation of sleep metabolism interactions. Taken together, a mechanistic understanding of how different genes function to regulate sleep in flies will further our understanding of how sleep and metabolism is regulated in humans. / Includes bibliography. / Dissertation (PhD)--Florida Atlantic University, 2021. / FAU Electronic Theses and Dissertations Collection

The clinical practice guideline (CPG) is the implementation of a depression screening tool to be used in the early perinatal period. The practice change occurred in a rural Obstetrical and Gynecological (OBGYN) practice in the southern United States. The CPG change has been guided by recommendations from both the American Congress of Obstetrics and Gynecology and the American College of Nurse Midwives. Implementation of this CPG change addresses the gap in practice of not doing depression screening during the perinatal period, and only screening during the postpartum period, which was being done at the OBGYN office. Theorist Lewin’s ‘change theory’ guided the implementation of the project. In order to apply this project, a process of changing practice guidelines was needed at the OBGYN office. The DNP project presented the practice change guideline of implementing the Edinburgh Postpartum Depression Screening (EPDS) tool. The need for the CPG development was evaluated by 3 nurse leaders using the AGREE II tool and was recommended 100% without modifications by all 3 evaluators. The CPG, with the results, was presented and discussed with the practice site’s practitioners. The practitioners implemented the EPDS to be given at the 12- week checkup appointment versus the confirmation of pregnancy appointment, which was suggested through the DNP project’s CPG. The implementation of this CPG has the potential to provide a safer environment for pregnant women, their newborns, and their families.