A Continuous, Fluorescence-based Assay of µ-Opioid Receptor Activation in AtT-20 Cells

Opioids are widely prescribed analgesics, but their use is limited due to development of tolerance and addiction, as well as high variability in individual response. The development of improved opioid analgesics requires high-throughput functional assays to assess large numbers of potential opioid ligands. In this study, we assessed the ability of a proprietary "no-wash" fluorescent membrane potential dye to act as a reporter of µ-opioid receptor (MOR) activation and desensitization via activation of G-protein-coupled inwardly rectifying potassium channels. AtT-20 cells stably expressing mouse MOR were assayed in 96-well plates using the Molecular Devices FLIPR membrane potential dye. Dye emission intensity decreased upon membrane hyperpolarization. Fluorescence decreased in a concentration-dependent manner upon application of a range of opioid ligands to the cells, with high-efficacy agonists producing a decrease of 35% to 40% in total fluorescence. The maximum effect of morphine faded in the continued presence of agonist, reflecting receptor desensitization. The effects of opioids were prevented by prior treatment with pertussis toxin and blocked by naloxone. We have demonstrated this assay to be an effective method for assessing ligand signaling at MOR, which may potentially be scaled up as an additional high-throughput screening technique for characterizing novel opioid ligands.NHMRC Grant Numbers: 1011979 & 104596

Assessment of Longitudinal Efficacy of Community Based Intervention for Schistosomiasis in Mara District, Lake Victoria Region of Tanzania

Program/Project Purpose: An estimated 200 million people are infected with schistosomiasis, making the disease one of the top neglected tropical diseases and a major global health concern. In Tanzania, with a population just over 42 million, about 33 million are in need of treatment or preventative chemotherapy. The disease is of particular concern in the Lake Victoria region, due to daily use of lake water. In conjunction with the Shirati district hospital, Touro University-CA has led control programs carried out in mobile clinics at local villages by providing screening, treatment, and education. Recently there has been a push for use of static dispensary locations for diagnosis and treatment. The aim of this study is to assess the longitudinal efficacy of community based interventions for schistosomiasis carried out from 2009-2014 and compares the utilization of static health dispensary clinics to the community based mobile clinics in reaching out to the population. Structure/Method/Design: Villagers were alerted to the screening, treatment, and education program through local leaders, announcements over the radio, and posters. Effectiveness will be measured by analyzing trends in the number of people visiting the clinics, prevalence of infection, the number treated, given positive infection status, and change in prevalence patterns with treatment. The number of people screened at dispensaries versus mobile clinics will be compared. We hypothesize that mobile clinics are effective in reducing prevalence of schistosomiasis in community based interventions and that health dispensaries provide no additional value towards increasing number of individuals screened. Outcomes & Evaluation: Univariate analysis and 2-tailed t-tests will be performed to assess if the use of services has significantly increased or not at mobile clinics from 2009-2014, and to compare utilization of health dispensaries versus mobile clinics. Prevalence of those who test positive will also be determined and analyzed to assess for a downward trend. Preliminary analysis reveals that from 2012 to 2014, prevalence of infection has decreased and that utilization of health dispensaries provides no additional value over mobile clinics. Our study supports the use of mobile clinics to carry out community-based health education, screening, and treatment programs over the use of dispensaries in rural Tanzania. Going Forward: During this study mobile clinics reported decreased prevalence and are able to capture a larger population of people interested in being screened for infection. Since use of dispensaries is favored by the Mara district, it will be important to point out the efficiency of mobile clinics or improve the efficacy of dispensaries by increasing the frequency of screenings or widening the area of promotion to other nearby villages

Providing an Oral Examination as an Authentic Assessment in a Large Section, Undergraduate Diversity Class

This study reflects on the development and implementation of mid-term oral examinations in large-scale lecture courses at a large, public research university; specifically, this work examines the implications of oral exams for fostering student engagement and concept-based comprehension in addition to institutional and course commitments to diversity. This research traces the development of an effective method for administering oral midterms and assesses the advantages and challenges of utilizing oral examinations for student assessment by detailing student feedback and TAs’ reactions to administering this examination format. Findings reveal that oral examinations provided a chance for students to develop skills through a different means of engaging material and to foster a concept-based learning approach. In a discussion of student and TA reactions, this paper reports a predominantly positive assessment by both groups while noting the challenges and disadvantages of this format

Pathogenic Mechanisms and Host Interactions in Staphylococcus epidermidis Device-Related Infection

Staphylococcus epidermidis is a permanent member of the normal human microbiota, commonly found on skin and mucous membranes. By adhering to tissue surface moieties of the host via specific adhesins, S. epidermidis is capable of establishing a lifelong commensal relationship with humans that begins early in life. In its role as a commensal organism, S. epidermidis is thought to provide benefits to human host, including out-competing more virulent pathogens. However, largely due to its capacity to form biofilm on implanted foreign bodies, S. epidermidis has emerged as an important opportunistic pathogen in patients receiving medical devices. S. epidermidis causes approximately 20% of all orthopedic device-related infections (ODRIs), increasing up to 50%in late-developing infections. Despite this prevalence, it remains underrepresented in the scientific literature, in particular lagging behind the study of the S. aureus. This review aims to provide an overview of the interactions of S. epidermidis with the human host, both as a commensal and as a pathogen. The mechanisms retained by S. epidermidis that enable colonization of human skin as well as invasive infection, will be described, with a particular focus upon biofilm formation. The host immune responses to these infections are also described, including how S. epidermidis seems to trigger low levels of pro-inflammatory cytokines and high levels of interleukin-10, which may contribute to the sub-acute and persistent nature often associated with these infections. The adaptive immune response to S. epidermidis remains poorly described, and represents an area which may provide significant new discoveries in the coming years

Production of ritual material culture in the Pre-Pottery Neolithic period in Jordan : some methods for analytical investigation

The Neolithic period in Jordan has come under increasing study since the accidental discovery of 'Ain Ghazal in the late 1970s, which not only showed that large settlements were present in marginal areas, but proved to be the type site of a sub-period previously unknown in the archaeological record. As more sites are discovered and excavated, prior entrenched notions of site interactions, demographic and settlement change, ritual, and other aspects of social organization have been questioned. The discovery of both large, densely occupied "mega-sites" and smaller, perhaps more specialized, peripheral sites which were apparently totally independent of the larger sites in the Pre-Pottery Neolithic has led to the idea of a polycentric evolution of Neolithic ideas and lifeways. However, while most researchers believe that these sites were self-sufficient, this does not preclude their interaction with other sites. In this thesis, potential ritual remains from Tel Tifdan, one of these small peripheral sites in Southern Jordan, are analyzed using chemical analyses to determine whether evidence of ritual trade with nearby or distant sites can be ascertained. X- Ray Fluorescence (XRF) and Fourier-Transform Infrared (FTIR) spectroscopy are employed to determine whether the clay from a sample of figurines from the site was sourced locally. The results are inconclusive as far as location of clay sources, but theoretically could be solidified with analysis using different chemical method

Production of ritual material culture in the Pre-Pottery Neolithic period in Jordan : some methods for analytical investigation

The Neolithic period in Jordan has come under increasing study since the accidental discovery of 'Ain Ghazal in the late 1970s, which not only showed that large settlements were present in marginal areas, but proved to be the type site of a sub-period previously unknown in the archaeological record. As more sites are discovered and excavated, prior entrenched notions of site interactions, demographic and settlement change, ritual, and other aspects of social organization have been questioned. The discovery of both large, densely occupied "mega-sites" and smaller, perhaps more specialized, peripheral sites which were apparently totally independent of the larger sites in the Pre-Pottery Neolithic has led to the idea of a polycentric evolution of Neolithic ideas and lifeways. However, while most researchers believe that these sites were self-sufficient, this does not preclude their interaction with other sites. In this thesis, potential ritual remains from Tel Tifdan, one of these small peripheral sites in Southern Jordan, are analyzed using chemical analyses to determine whether evidence of ritual trade with nearby or distant sites can be ascertained. X- Ray Fluorescence (XRF) and Fourier-Transform Infrared (FTIR) spectroscopy are employed to determine whether the clay from a sample of figurines from the site was sourced locally. The results are inconclusive as far as location of clay sources, but theoretically could be solidified with analysis using different chemical method

Providing an Oral Examination as an Authentic Assessment in a Large Section, Undergraduate Diversity Class

This study reflects on the development and implementation of mid-term oral examinations in large-scale lecture courses at a large, public research university; specifically, this work examines the implications of oral exams for fostering student engagement and concept-based comprehension in addition to institutional and course commitments to diversity. This research traces the development of an effective method for administering oral midterms and assesses the advantages and challenges of utilizing oral examinations for student assessment by detailing student feedback and TAs’ reactions to administering this examination format. Findings reveal that oral examinations provided a chance for students to develop skills through a different means of engaging material and to foster a concept-based learning approach. In a discussion of student and TA reactions, this paper reports a predominantly positive assessment by both groups while noting the challenges and disadvantages of this format