Investigations into the function of Elp3 in Toxoplasma gondii

Indiana University-Purdue University Indianapolis (IUPUI)The parasite Toxoplasma gondii causes life-threatening infection in immunocompromised individuals. Our lab has determined that Toxoplasma Elongator protein-3 (TgElp3) is required for parasite viability. While catalytic domains are conserved, TgElp3 is the only component of the six-subunit Elongator complex present in Toxoplasma; moreover, TgElp3 localizes to the outer mitochondria membrane (OMM). These unusual features suggest that TgElp3 may have unique roles in parasite biology that could be useful in drug targeting. The goals of this thesis were to determine the function of TgElp3 and how the protein traffics to the OMM. In other species, Elp3 mediates lysine acetylation of histones and alphatubulin, and its radical S-adenosyl methionine (rSAM) domain is important for the formation of tRNA modifications, which enhance translation efficiency and fidelity. Given its location, histones would not be an expected substrate, and we further determined that tubulin acetylation in Toxoplasma is mediated by a different enzyme, TgATAT. We found that overexpression of TgElp3 at the parasite’s mitochondrion results in a significant replication defect, but overexpression of TgElp3 lacking the transmembrane domain (TMD) or with a mutant rSAM domain is tolerated. We identified one such modification, 5-methoxycarbonylmethyl-2thiouridine (mcm5S2U) that is likely mediated by TgElp3. These findings signify the importance of TgElp3’s rSAM domain for protein function, and confirms TgElp3 activity at the OMM is essential for Toxoplasma viability as previously reported. To determine how TgElp3 traffics to the OMM, we performed a bioinformatics survey that discovered over 50 additional “tail-anchored” proteins present in Toxoplasma. Mutational analyses found that targeting of these TA proteins to specific parasite organelles was strongly influenced by the TMD sequence, including charge of the flanking C-terminal sequence

The influence of pulmonary rehabilitation and counselling on perceptions of physical activity in individuals with COPD : a qualitative study

Objective: The aim of this study was to explore to what extent a combined counselling and pulmonary rehabilitation program (PR) influences the perception of physical activity (PA) and motivation for behavioral change in PA in individuals with COPD. The results of previous quantitative trial that investigated the effect of this combined treatment on daily PA were inconclusive. It is conjectured that a more targeted tailoring of the counselling and PR intervention could improve its effectiveness. Patients and Methods: Eighteen individuals with COPD (median age 69, 8 females) who had participated in the PneumoReha program were interviewed twice (following PR and at three-month follow-up). These interviews were transcribed and analyzed thematically. Based on the codes thus identified, three categories ‘perception of PA intensity’, ‘quality of motivation to perform PA’, and ‘strategies to cope with barriers’ were used to differentiate ‘types’ of participants. Results: Four different types of COPD individuals were distinguished. Study findings indicate that those individuals who participated in the PR program combined with embedded counselling tended to be more active and intrinsically motivated. Conclusion: A typology of four types of people with COPD was developed, characterized by their perception of activity, individual motivation and strategies for managing barriers. The patients’ physical activity level might be influenced by their concept of physical activity and the quality of motivation. Recognizing patients’ different activity behaviors is important for improving the quality of outpatient PR programs and developing tailored (according to each type) counselling interventions embedded in outpatient PR programs. Clinical Trial Registration: The study was registered on the website of https://www.clinicaltrials.gov/ with the identifier NCT02455206 (27/05/2015), as well as on the Swiss National Trails Portal SNCTP000001426 (05/21/2015)

Building More Inclusive Boards & Committees: Immigrant Participation in Government Decision-making

46 pagesLocal government agencies engage the public in decision-making to increase their understanding of local issues, make better decisions, and build trust along the way. One common tactic for sustained engagement are “citizen” advisory boards and committees. This research is a part of a broader effort, led by Professor José W. Meléndez, to inventory a cross-section of these boards and committees in Oregon to better understand the current state of representation. The research team documented the incorporation of immigrants in local and state government boards and interviewed nearly 50 first- and second-generation immigrants. The research addresses three questions: 1) What are the different levels of board decision-making available across the state? 2) What is the state of immigrant representation within these roles? And 3) What barriers do immigrants face in accessing decision-making roles? My research targets three types of boards that influence decision-making across the state. Immigrant participants represent a variety of local and statewide groups across 1) school district boards, which serve as governing bodies with broad decision-making authority; 2) budget committees, which are statutory and work directly with local elected officials; and 3) transportation advisory bodies with more limited influence. Through the analysis of 15 interviews across these three types of bodies, I answer a final research question: how can government institutions expand and support the inclusion of immigrants in decision-making roles? This report shares findings and suggested strategies to increase inclusiveness on government boards and committees

Physiotherapists’ use and perceptions of digital remote physiotherapy during COVID-19 lockdown in Switzerland ::an online cross-sectional survey

Background : The Swiss containment strategy for the COVID-19 pandemic during the first wave in spring 2020 resulted in a moratorium on non-urgent physiotherapy via regular direct patient contact. Consequently, such physiotherapy sessions declined by 84%. This study investigates the impact of this moratorium on the use of digital remote physiotherapy in Switzerland during this period and the perceptions of its use by Swiss physiotherapists (PTs). Methods: A cross-sectional online questionnaire was distributed between June and August of 2020 via the Swiss Physiotherapy Association (physioswiss) and various associations of physiotherapy specialists (e.g., sport, pediatric) working in both inpatient and outpatient settings. The questionnaire was designed to capture the demographics of participants and the perceptions of PTs using 33 questions in the following domains: Demography; Attitudes towards digital technology; Private and professional use of digital technology; Use of digital technology during therapy; and, Support requirements. Closed and open-ended questions were included and the frequency of answers was analyzed. Non-parametric inferential statistics were used to identify differences, where appropriate. The Checklist for Reporting Results of Internet E-Surveys (CHERRIES) was adopted. Results: Participants in the survey were 742 PTs (23.5% male, mean age of 43 years, mean working experience of 18 years) from the German-speaking (75.5%), French-speaking (15.1%), and Italian-speaking (9.4%) regions of Switzerland. The percentage of PTs using digital remote therapy increased from 4.9% prior to the lockdown to 44.6% during the lockdown period. The majority of PTs did not consider that digital remote therapy could complement usual physiotherapy practice and did not plan to continue with digital remote therapy after the pandemic. Conclusions: During the lockdown, Swiss PTs adopted various low-cost and easily accessible digital technologies. However, several barriers hampered further implementation of this modality. Specific education and training programs need to be provided among PTs, appropriate digital technologies should be introduced, and a correct reimbursement scheme should be developed. Trial registration: COVIDPhysio Registry of World Physiotherapy, registered 15th June 2020 (https://world.physio/covid-19-information-hub/covid-19-covidphysio-registry)

“A State of Immigrants”: A New Look at the Immigrant Experience in Oregon

155 pagesEn 2008, docentes de la Universidad de Oregón publicaron un reporte titulado “La experiencia de los inmigrantes en Oregón.” Ya que la población inmigrante y refugiada en Oregón ha presentado un mayor aumento desde la década del 1990, les autores buscaban ampliar el conocimiento público sobre la experiencia de les inmigrantes en Oregón, documentar las contribuciones de les inmigrantes tanto en la comunidad como en los espacios de trabajo, y ofrecer recomendaciones que ayudarían a un exitoso proceso de integración de les inmigrantes a la vida social, económica y cívica del estado. El informe se difundió ampliamente y generó un debate reflexivo sobre el papel vital de las inmigrantes en el estado y la necesidad de generar acciones y políticas que aborden sus necesidades e intereses

“A State of Immigrants”: A New Look at the Immigrant Experience in Oregon

136 pagesThis work was funded by the Andrew W. Mellon Foundation

Landscape of Preterm Birth Therapeutics and a Path Forward

Preterm birth (PTB) remains the leading cause of infant morbidity and mortality. Despite 50 years of research, therapeutic options are limited and many lack clear efficacy. Tocolytic agents are drugs that briefly delay PTB, typically to allow antenatal corticosteroid administration for accelerating fetal lung maturity or to transfer patients to high-level care facilities. Globally, there is an unmet need for better tocolytic agents, particularly in low- and middle-income countries. Although most tocolytics, such as betamimetics and indomethacin, suppress downstream mediators of the parturition pathway, newer therapeutics are being designed to selectively target inflammatory checkpoints with the goal of providing broader and more effective tocolysis. However, the relatively small market for new PTB therapeutics and formidable regulatory hurdles have led to minimal pharmaceutical interest and a stagnant drug pipeline. In this review, we present the current landscape of PTB therapeutics, assessing the history of drug development, mechanisms of action, adverse effects, and the updated literature on drug efficacy. We also review the regulatory hurdles and other obstacles impairing novel tocolytic development. Ultimately, we present possible steps to expedite drug development and meet the growing need for effective preterm birth therapeutics

Combined HIV-1 sequence and integration site analysis informs viral dynamics and allows reconstruction of replicating viral ancestors

Understanding HIV-1 persistence despite antiretroviral therapy (ART) is of paramount importance. Both single-genome sequencing (SGS) and integration site analysis (ISA) provide useful information regarding the structure of persistent HIV DNA populations; however, until recently, there was no way to link integration sites to their cognate proviral sequences. Here, we used multiple-displacement amplification (MDA) of cellular DNA diluted to a proviral endpoint to obtain full-length proviral sequences and their corresponding sites of integration. We applied this method to lymph node and peripheral blood mononuclear cells from 5 ART-treated donors to determine whether groups of identical subgenomic sequences in the 2 compartments are the result of clonal expansion of infected cells or a viral genetic bottleneck. We found that identical proviral sequences can result from both cellular expansion and viral genetic bottlenecks occurring prior to ART initiation and following ART failure. We identified an expanded T cell clone carrying an intact provirus that matched a variant previously detected by viral outgrowth assays and expanded clones with wild-type and drug-resistant defective proviruses. We also found 2 clones from 1 donor that carried identical proviruses except for nonoverlapping deletions, from which we could infer the sequence of the intact parental virus. Thus, MDA-SGS can be used for "viral reconstruction" to better understand intrapatient HIV-1 evolution and to determine the clonality and structure of proviruses within expanded clones, including those with drug-resistant mutations. Importantly, we demonstrate that identical sequences observed by standard SGS are not always sufficient to establish proviral clonality