Investigating the Role of Vaccinia virus Immune Suppressing Protein B19 in Aiding Function of E3 in Pathogenesis

135663-Thumbnail Image.png
Description
Vaccinia virus (VV) is a prototype virus of the Orthopox viruses. The large dsDNA virus composed of 200kbp genome contains approximately 200 genes and replicates entirely in the cytosol. Since its use as a live vaccine against smallpox that leads

Vaccinia virus (VV) is a prototype virus of the Orthopox viruses. The large dsDNA virus composed of 200kbp genome contains approximately 200 genes and replicates entirely in the cytosol. Since its use as a live vaccine against smallpox that leads to the successful eradication of smallpox, Vaccinia has been intensely studied as a vaccine vector since the large genome allows for the insertion of multiple genes. It is also studied as a molecular tool for gene therapy and gene functional study. Despite its success as a live vaccine, the vaccination causes some mild to serious bur rare adverse events in vaccinees such as generalized Vaccinia and encepharitis. Therefore, identification of virulence genes and removal of these genes to create a safer vaccine remain an important tasks. In this study, the author seeks to elucidate the possible relationship between immune evading proteins E3 and B19. VV did not allow double deletions of E3 and B19, indicating the existence of a relationship between the two genes.
Date Created
2016-05
Agent

Analysis of an HIV/AIDS Prevention Education Program for Secondary Students in Tanzania

134992-Thumbnail Image.png
Description
The purpose of this study was to evaluate the efficacy and quality of HEAL International's HIV/AIDS education prevention program for secondary school students in the Arusha region of Tanzania during the summer of 2016 using a cross-cultural teaching team. Basic

The purpose of this study was to evaluate the efficacy and quality of HEAL International's HIV/AIDS education prevention program for secondary school students in the Arusha region of Tanzania during the summer of 2016 using a cross-cultural teaching team. Basic HIV/AIDS knowledge and attitudes concerning risk reduction behaviors as well as towards people living with HIV/AIDS were studied among Form 1 and Form 3 students from two secondary schools in rural Tanzania. The intervention program aimed to increase knowledge and positive attitudes related to HIV/AIDS in order to motivate healthy behavior change. 211 Form 1 students and 156 Form 3 students received the intervention and completed both pre- and post-evaluation surveys. At the post-evaluation, all students showed increases in basic HIV/AIDS knowledge levels as well as positive attitudes concerning HIV/AIDS risk reduction and about people living with HIV/AIDS. Students' levels of uncertainty when answering the survey questions were also decreased. Overall, the study findings indicate that HEAL's program had a positive impact on HIV/AIDS related knowledge and attitudes of secondary school students in Arusha, Tanzania. While this study had many limitations, it also offers areas of improvement for future HEAL International volunteer programs.
Date Created
2016-12
Agent

Understanding Bioethical Concerns of Laboratory Research: A Case Study On Discovering Programmed Necrosis Pathways

134904-Thumbnail Image.png
Description
The concept of “good” research is concrete in terms of technique, but complex in theory. As technology advances, the complexity of problems we must solve also grows. Research is facing an ethical dilemma—which projects, applied or basic, should be funded.

The concept of “good” research is concrete in terms of technique, but complex in theory. As technology advances, the complexity of problems we must solve also grows. Research is facing an ethical dilemma—which projects, applied or basic, should be funded. Research is no longer an isolated sector in society, and the decisions made inside of the laboratory are affecting the general public more directly than ever before. While there is no correct answer to what the future of research should be, it is clear that good research can no longer be only defined by the current classification system, which is rooted in antiquated, yet ingrained, social status distinctions.
Differences between basic and applied research were explored through a wet-lab case study. Vaccinia virus (VACV) infections are a prime model of the competition between a virus and its host. VACV contains a gene that is highly evasive of the host immune system, gene E3L. The protein encoded by E3L is E3, which contains two highly conserved regions, a C-terminus, and a N-terminus. While the C-terminus is well-understood, the mechanism by which the N-terminus grants IFN resistance was previously unknown. This project demonstrated that the N-terminus prevents the initiation of programmed necrosis through host-encoded cellular proteins RIP3 and DAI. These findings provide insight into the function of the N-terminus of E3, as well as the unique functions of induced programmed necrosis.
This project was an example of “basic” research. However, it highlights the interconnectivity of basic and applied research and the danger in isolating both projects and perspectives. It points to the difficult decisions that must be made in science, and the need for a better research classification system that considers what makes science “good” outside of antiquated social class ideologies that have shaped science since ancient Greece. While there are no easy answers to determine what makes research “good,” thinking critically about the types of research projects that will be pursued, and the effects that research has on both science and society, will raise awareness, initiate new conversations, and encourage more dialogue about science in the 21st century.
Date Created
2016-12
Agent

Inhibition of PKR phosphorylation by Vaccinia Virus' E3 Protein

134414-Thumbnail Image.png
Description
Vaccinia virus is a cytoplasmic, double-stranded DNA orthopoxvirus. Unlike mammalian cells, vaccinia virus produces double-stranded RNA (dsRNA) during its viral life cycle. The protein kinase R, PKR, is one of the principal host defense mechanisms against orthopoxvirus infection. PKR can

Vaccinia virus is a cytoplasmic, double-stranded DNA orthopoxvirus. Unlike mammalian cells, vaccinia virus produces double-stranded RNA (dsRNA) during its viral life cycle. The protein kinase R, PKR, is one of the principal host defense mechanisms against orthopoxvirus infection. PKR can bind double-stranded RNA and phosphorylate eukaryotic translation initiation factor, eIF2α, shutting down protein synthesis and halting the viral life cycle. To combat host defenses, vaccinia virus encodes E3, a potent inhibitor of the cellular anti-viral eIF2α kinase, PKR. The E3 protein contains a C-terminal dsRNA-binding motif that sequesters dsRNA and inhibits PKR activation. We demonstrate that E3 also interacts with PKR by co-immunoprecipitation. This interaction is independent of the presence of dsRNA and dsRNA-binding by E3, indicating that the interaction is not due to dsRNA-bridging.
PKR interaction mapped to a region within the dsRNA-binding domain of E3 and overlapped with sequences in the C-terminus of this domain that are necessary for binding to dsRNA. Point mutants of E3 were generated and screened for PKR inhibition and direct interaction. Analysis of these mutants demonstrates that dsRNA-binding but not PKR interaction plays a critical role in the broad host range of VACV. Nonetheless, full inhibition of PKR in cells in culture requires both dsRNA-binding and PKR interaction. Because E3 is highly conserved among orthopoxviruses, understanding the mechanisms that E3 uses to inhibit PKR can give insight into host range pathogenesis of dsRNA producing viruses.
Date Created
2017-05
Agent

Proposal to Institute a Needle Exchange in Maricopa County

137595-Thumbnail Image.png
Description
Utilizing a compilation of existing literature, theory and current praxis, a clear case for instituting a needle exchange in Maricopa County is visible.. Research on HIV rates are consulted to establish a foundation for understanding the value of needle exchange

Utilizing a compilation of existing literature, theory and current praxis, a clear case for instituting a needle exchange in Maricopa County is visible.. Research on HIV rates are consulted to establish a foundation for understanding the value of needle exchange programs nationally and in Maricopa County. Case studies on existing needle exchange efficacy are also used to support the proposal and outline the success of needle exchanges in reducing rates of infection.
Date Created
2013-05
Agent

An Evaluation of HEAL International's Health Education Program in Tanzania, Africa

137173-Thumbnail Image.png
Description
This study investigated the potential efficacy of HEAL International's prevention education program in inducing health behavior change in HIV/AIDS, malaria, and communicable disease to children in grade levels ranging from primary school to secondary school. The health education program was

This study investigated the potential efficacy of HEAL International's prevention education program in inducing health behavior change in HIV/AIDS, malaria, and communicable disease to children in grade levels ranging from primary school to secondary school. The health education program was aimed at changing health behavior by increasing knowledge. This increase in knowledge was analyzed as a modifying factor in the Health Belief Model suggesting that knowledge, along with five other modifying factors, are directly responsible for an individual's health perceptions. These health perceptions ultimately result in an individual's health behavior. As a result, it is argued that an increase in knowledge can lead to health behavior change so long as it is coupled with a strong theoretical framework. Administering pre-evaluations at the beginning of the program, post evaluations at the end of the program, and a second post evaluation again two months later completed the evaluation. It was hypothesized that if there was a significant difference between the percent of correct answers at the pre-evaluation compared the second post-evaluation then there is evidence that HEAL's health education program is, or at least has the potential to, create sustainable health behavior change. A paired samples t-test was completed on the data and showed a statistically significant difference between the percent of correct answers at pre-evaluation and the percent of correct answers at second post-evaluation. These results indicated that the number of students with a comprehensive knowledge of the subjects that HEAL taught during the program had increased. It was concluded that the results of the study indicate evidence that HEAL's program has the potential to deliver sustainable health behavior change but that it will be more quantifiable once HEAL is able to adopt a theoretical framework on which to base future programs.
Date Created
2014-05
Agent

STUDY OF INACTIVATION OF MS2 BACTERIOPHAGE BY ULTRA SHORT PULSES OF TI-SAPPHIRE LASER

Description
Studies have demonstrated that viruses such as human immunodeficiency virus [HIV], M13 bacteriophage, and murine cytomegalovirus [MCMV] have been effectively inactivated by exposure to ultra short-pulsed lasers (6,7,10,11,14,15,17). Ultra short pulse laser shows promise as a new method for non-invasive

Studies have demonstrated that viruses such as human immunodeficiency virus [HIV], M13 bacteriophage, and murine cytomegalovirus [MCMV] have been effectively inactivated by exposure to ultra short-pulsed lasers (6,7,10,11,14,15,17). Ultra short pulse laser shows promise as a new method for non-invasive antiviral treatments (17). This method can be used to prevent problems such as drug resistance that is currently rising in numbers. According to the Center for Disease Control [CDC], there are more than two million people in the United States of America that are infected with antimicrobial-resistant infections and at least 23,000 deaths per year occur as a result (19). In this study, ultra-short pulses, specifically Ti-Sapphire Laser [USP Ti-Sapphire Laser] will be evaluated for viral inactivation. The virus chosen for this study was MS2 bacteriophage, which is a non- enveloped, icosahedral, single-stranded RNA [ssRNA] bacteriophages that infects F+ pilus Escherichia coli (16). It was hypothesized that ultrashort pulses from a Ti-Sapphire laser will inactivate MS2 bacteriophage. Inactivation was measured using plaque-forming units [PFU/mL] as an indicator. It was expected that there would be an increase in inactivation of MS2 bacteriophage with an increase in irradiation duration. The results indicated that MS2 bacteriophage was highly sensitive to irradiation treatments of the USP Ti-Sapphire Laser. The concentration of MS2 bacteriophage decreased by 107 PFU/mL after being treated for various time periods ranging from 5 minutes to 150 minutes. Longer duration of USP Ti- Sapphire Laser treatment inactivated more MS2 Bacteriophage.
Date Created
2014-05
Agent

Describing Virus Vaccines and Controversies: Comparison of Three Vaccines in Medical and Social Contexts

136878-Thumbnail Image.png
Description
In the United States, a dispute has arisen over the safety and need for vaccination, particularly in regard to compulsory vaccination laws. New outlets and social media sites publish countless reports about the dangers of vaccines or of known adverse

In the United States, a dispute has arisen over the safety and need for vaccination, particularly in regard to compulsory vaccination laws. New outlets and social media sites publish countless reports about the dangers of vaccines or of known adverse reactions as well as imagined or unproven worries. Individuals' rights to choose to get vaccinated or allow their children to be vaccinated comes to direct conflict with measures needed to protect communities from preventable viral diseases. The controversy surrounding vaccines is not new, nor necessarily are the fundamental reasons for skepticism. Looking back through the history of vaccines as a medical tool, the evolution of the controversy can be observed taking place with each new historical context, scientific development, and social conditions. Despite scientific research and assurances of vaccine safety, opposition and unease about vaccination appear to take Looking individually at the development and distribution of the smallpox (variola virus), polio (poliovirus) and human papilloma virus(HPV) vaccines, concerns regarding the violation of personal rights, safety of vaccines themselves, and social stigmas and connotations surrounding vaccines can be seen to evolve and change. Due to the way doubt can manifest in different ways over time, it may be impossible to fully end the vaccine debate. However, nderstanding the sociological factors behind anti-vaccine sentiment may allow healthcare professionals to work with concerned people with a particular care to address these visceral and sometimes irrational fears surrounding vaccination.
Date Created
2014-05
Agent

Importance of Cholesterol Depletion in Measles Virus Infection and Replication

136311-Thumbnail Image.png
Description
Lipid membranes are a key structure for many classes of viruses. Lipid membranes can be analyzed using the fluid mosaic model, which states that the phospholipid membrane has variable amounts of fluidity and key membrane proteins are presented in areas

Lipid membranes are a key structure for many classes of viruses. Lipid membranes can be analyzed using the fluid mosaic model, which states that the phospholipid membrane has variable amounts of fluidity and key membrane proteins are presented in areas stabilized by cholesterol-enriched platforms called lipid rafts. This project aims to further the understanding of the importance of lipid rafts in measles virus (MV) infection and replication, which has not been extensively studied. In order to do this, an MV-susceptible cell line was treated with an anti-cholesterol compound before and after measles virus infection. I found that pre-infection treatments had a marginal effect upon measles cytopathic effect (syncytia formation) or replication. Twenty-four hours post-infection treatment had a deleterious effect on cell viability, but the replication/assembly of infectious units per cell decreased importantly and in dose-dependent manner. Furthermore, by measuring the susceptibility to neutralization of infectious particles obtained from MBCD treated cells, I determined the importance of lipid microdomain environment on the stability of infectious particles. Increased anti-cholesterol treatment enhanced the susceptibility of MV to neutralization. Future studies are proposed to assess the properties of cholesterol depleted viral infectious units.
Date Created
2015-05
Agent

Characterization of the Physiological Response Following In Vivo Administration of Astragalus Membranaceus

128615-Thumbnail Image.png
Description

The botanical, Astragalus membranaceus, is a therapeutic in traditional Chinese medicine. Limited literature exists on the overall in vivo effects of A. membranaceus on the human body. This study evaluates the physiological responses to A. membranaceus by measuring leukocyte, platelet,

The botanical, Astragalus membranaceus, is a therapeutic in traditional Chinese medicine. Limited literature exists on the overall in vivo effects of A. membranaceus on the human body. This study evaluates the physiological responses to A. membranaceus by measuring leukocyte, platelet, and cytokine responses as well as body temperature and blood pressure in healthy individuals after the in vivo administration of A. membranaceus. A dose-dependent increase in monocytes, neutrophils, and lymphocytes was measured 8–12 hours after administration and an increase in the number of circulating platelets was seen as early as 4 hours. A dynamic change in the levels of circulating cytokines was observed, especially in interferon-γ and tumor necrosis factor-α, IL-13, IL-6, and soluble IL-2R. Subjective symptoms reported by participants were similar to those typically experienced in viral type immune responses and included fatigue, malaise, and headache. Systolic and diastolic blood pressure were reduced within 4 hours after administration, while body temperature mildly increased within 8 hours after administration. In general, all responses returned to baseline values by 24 hours. Collectively, these results support the role of A. membranaceus in priming for a potential immune response as well as its effect on blood flow and wound healing.

Date Created
2016-03-30
Agent