Matching Items (62)
Description
Human papillomavirus (HPV) is the causative agent of cervical cancer. Persistent infection with high-risk HPV 16, 18 or 45 species is associated with the development and progression of cervical cancer. HPV genotyping and Pap smear tests are the regular methods used to detect pre-invasive cervical lesions, but there is a need for developing a rapid biomarker to profile immunity to these viruses. The viral E7 oncogene is expressed in most HPV-associated cancers and anti-E7 antibodies can be detected in the blood of patients with cervical cancer. This research was focused on viral E7 oncogene expression to be used in development of low-cost point of care tests, enabling patients from low resource settings to detect the asymptotic stage of cervical cancer and be able to seek treatment early. In order to produce the E7 protein in vitro to measure antibody levels, GST tagged E7 genes from HPV 16, 18 and 45 species were inserted into the pDEST15 vector and expressed in E. coli BL21DE3 cells that were induced with 1mM of IPTG. The E7-GST fused expressed protein was then purified using glutathione beads and resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Protein expression was 5.8 \u03bcg/ml for HPV 16E7 in 500 ml culture and for the 500 ml culture of HPV 18 E7 and 45 E7 were 10.5 \u03bcg/ml and 10.5 \u03bcg/ml for HPV 18E7 and 45E7 respectively. High yield values are showing high expression levels of GST-tagged E7 recombinant protein which can be used for serotyping a number of individuals. This shows that HPV E7 can be produced in large quantities that can potentially be used in point of care tests that can help identify women at risk of cervical cancer. In conclusion, the E7 protein produced in this study can potentially be used to induce humoral responses in patients\u2019 sera for understanding the immune response of cervical cancer.
ContributorsMakuyana, Ntombizodwa (Author) / Anderson, Karen (Thesis director) / Ewaisha, Radwa (Committee member) / Varsani, Arvind (Committee member) / Hou, Ching-Wen (Committee member) / School of Mathematical and Statistical Sciences (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2018-12
Description
In response to a national call within STEM to increase diversity within the sciences, there has been a growth in science education research aimed at increasing participation of underrepresented groups in science, such as women and ethnic/racial minorities. However, an underexplored underrepresented group in science are religious students. Though 82% of the United States population is religiously affiliated, only 52% of scientists are religious (Pew, 2009). Even further, only 32% of biologists are religious, with 25% identifying as Christian (Pew, 2009; Ecklund, 2007). One reason as to why Christian individuals are underrepresented in biology is because faculty may express biases that affect students' ability to persist in the field of biology. In this study, we explored how revealing a Christian student's religious identity on science graduate application would impact faculty's perception of the student during the biology graduate application process. We found that faculty were significantly more likely to perceive the student who revealed their religious identity to be less competent, hirable, likeable, and faculty would be less likely to mentor the student. Our study informs upon possible reasons as to why there is an underrepresentation of Christians in science. This further suggests that bias against Christians must be addressed in order to avoid real-world, negative treatment of Christians in science.
ContributorsTruong, Jasmine Maylee (Author) / Brownell, Sara (Thesis director) / Gaughan, Monica (Committee member) / Barnes, Liz (Committee member) / School of Life Sciences (Contributor) / W.P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
Learning student names has been promoted as an inclusive classroom practice, but it is unknown whether students value having their names known by an instructor. We explored this question in the context of a high-enrollment active-learning undergraduate biology course. Using surveys and semistructured interviews, we investigated whether students perceived that instructors know their names, the importance of instructors knowing their names, and how instructors learned their names. We found that, while only 20% of students perceived their names were known in previous high-enrollment biology classes, 78% of students perceived that an instructor of this course knew their names. However, instructors only knew 53% of names, indicating that instructors do not have to know student names in order for students to perceive that their names are known. Using grounded theory, we identified nine reasons why students feel that having their names known is important. When we asked students how they perceived instructors learned their names, the most common response was instructor use of name tents during in-class discussion. These findings suggest that students can benefit from perceiving that instructors know their names and name tents could be a relatively easy way for students to think that instructors know their names. Academic self-concept is one's perception of his or her ability in an academic domain compared to other students. As college biology classrooms transition from lecturing to active learning, students interact more with each other and are likely comparing themselves more to students in the class. Student characteristics, such as gender and race/ethnicity, can impact the level of academic self-concept, however this has been unexplored in the context of undergraduate biology. In this study, we explored whether student characteristics can affect academic self-concept in the context of a college physiology course. Using a survey, students self-reported how smart they perceived themselves in the context of physiology compared to the whole class and compared to the student they worked most closely with in class. Using logistic regression, we found that males and native English speakers had significantly higher academic self-concept compared to the whole class compared with females and non-native English speakers, respectively. We also found that males and non-transfer students had significantly higher academic self-concept compared to the student they worked most closely with in class compared with females and transfer students, respectively. Using grounded theory, we identified ten distinct factors that influenced how students determined whether they are more or less smart than their groupmate. Finally, we found that students were more likely to report participating less than their groupmate if they had a lower academic self-concept. These findings suggest that student characteristics can influence students' academic self-concept, which in turn may influence their participation in small group discussion.
ContributorsKrieg, Anna Florence (Author) / Brownell, Sara (Thesis director) / Stout, Valerie (Committee member) / Cooper, Katelyn (Committee member) / School of Life Sciences (Contributor) / School of Politics and Global Studies (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
Course-Based Undergraduate Research Experiences, or CUREs have become an increasingly popular way to integrate research opportunities into the undergraduate biology curriculum. Unlike traditional cookbook labs which provide students with a set experimental design and known outcome, CUREs offer students the opportunity to participate in novel and interesting research that is of interest to the greater biology community. While CUREs have been championed as a way to provide more students with the opportunity to experience, it is unclear whether students benefit differently from participating in different CURE with different structural elements. In this study we focused in on one proposed element of a CURE, collaboration, to determine whether student's perception of this concept change over the course of a CURE and whether it differs among students enrolled in different CUREs. We analyzed pre and post open-ended surveys asking the question "Why might collaboration be important in science?" in two CUREs with different structures of collaboration. We also compared CURE student responses to the responses of senior honors thesis students who had been conducting authentic research. Five themes emerged in response to students' conceptions of collaboration. Comparing two CURE courses, we found that students' conceptions of collaboration were varied within each individual CURE, as well as what students were leaving with compared to the other CURE course. Looking at how student responses compared between 5 different themes, including "Different Perspectives", "Validate/Verify Results", "Compare Results", "Requires Different Expertise", and "Compare results", students appeared to be thinking about collaboration in distinct different ways by lack of continuity in the amount of students discussing each of these among the classes. In addition, we found that student responses in each of the CURE courses were not significantly different for any of the themes except "Different Expertise" compared to the graduating seniors. However, due to the small (n) that the graduating seniors group had, 22, compared to each of the CURE classes composing of 155 and 98 students, this comparison must be taken in a preliminary manner. Overall, students thought differently about collaboration between different CUREs. Still, a gap filling what it means to "collaborate", and whether the structures of CUREs are effective to portray collaboration are still necessary to fully elaborate on this paper's findings.
ContributorsWassef, Cyril Alexander (Author) / Brownell, Sara (Thesis director) / Stout, Valerie (Committee member) / Cooper, Katelyn (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
Globally, about two-thirds of the population is latently infected with herpes simplex virus type 1 (HSV-1). HSV-1 is a large double stranded DNA virus with a genome size of ~150kbp. Small defective genomes, which minimally contain an HSV-1 origin of replication and packaging signal, arise naturally via recombination during viral DNA replication. These small defective genomes can be mimicked by constructing a bacterial plasmid containing the HSV-1 origin of replication and packaging signal, transfecting these recombinant plasmids into mammalian cells, and infecting with a replicating helper virus. The absence of most viral genes in the amplicon vector allows large pieces of foreign DNA (up to 150kbp) to be incorporated. The HSV-1 amplicon is replicated and packaged by the helper virus to form HSV-1 particles containing the amplicon DNA. We constructed a novel HSV-1 amplicon vector system containing lambda phage-derived attR sites to facilitate insertion of transgenes by Invitrogen Gateway recombination. To demonstrate that the amplicon vectors work as expected, we packaged the vector constructs expressing Emerald GFP using the replication-competent helper viruses OK-14 or HSV-mScartlet-I-UL25 in Vero cells and demonstrate that the vector stock can subsequently transduce and express Emerald GFP. In further work, we will insert transgenes into the amplicon vector using Invitrogen Gateway recombination to study their functionality.
ContributorsVelarde, Kimberly (Author) / Hogue, Ian B (Thesis advisor) / Manfredsson, Fredric (Committee member) / Sandoval, Ivette (Committee member) / Varsani, Arvind (Committee member) / Arizona State University (Publisher)
Created2021
Description
Caracals (Caracal caracal) are a felid species native to regions of southern Africa and western and central Asia. Despite their relatively high prevalence, the majority of research conducted on caracals has been undertaken on captive individuals, which encounter significantly different environments and exhibit different behaviors in comparison to caracals in the wild. Thereby, they likely have a vastly different virome. The goal of this study was to identify known and unknown DNA viruses associated with free-ranging caracals. Caracal fecal and organ samples were obtained from a caracal surveillance study undertaken in the Western Cape region of South Africa. Parasitic ticks found feeding on caracals were also obtained. Using a viral metagenomic informed approach, a novel circovirus (family Circoviridae) was detected and characterized in caracal fecal, kidney, spleen, and liver samples, as well as in ticks feeding on the caracals. To our knowledge, this is the first circovirus identified in caracals. The novel circovirus was determined to be closely related to a canine circovirus. These findings expand the knowledge of viral diversity and caracals and are greatly important to caracal conservation efforts as well as conservation efforts of other animals within their ecosystem.
ContributorsCollins, Courtney (Author) / Varsani, Arvind (Thesis director) / Dolby, Greer (Committee member) / Kraberger, Simona (Committee member) / Barrett, The Honors College (Contributor) / School of Molecular Sciences (Contributor)
Created2022-05
Description
Evolution is a key feature of undergraduate biology education: the AmericanAssociation for the Advancement of Science (AAAS) has identified evolution as one of
the five core concepts of biology, and it is relevant to a wide array of biology-related
careers. If biology instructors want students to use evolution to address scientific
challenges post-graduation, students need to be able to apply evolutionary principles to
real-life situations, and accept that the theory of evolution is the best scientific
explanation for the unity and diversity of life on Earth. In order to help students progress
on both fronts, biology education researchers need surveys that measure evolution
acceptance and assessments that measure students’ ability to apply evolutionary concepts.
This dissertation improves the measurement of student understanding and acceptance of
evolution by (1) developing a novel Evolutionary Medicine Assessment that measures
students’ ability to apply the core principles of Evolutionary Medicine to a variety of
health-related scenarios, (2) reevaluating existing measures of student evolution
acceptance by using student interviews to assess response process validity, and (3)
correcting the validity issues identified on the most widely-used measure of evolution
acceptance - the Measure of Acceptance of the Theory of Evolution (MATE) - by
developing and validating a revised version of this survey: the MATE 2.0.
ContributorsMisheva, Anastasia Taya (Author) / Brownell, Sara (Thesis advisor) / Barnes, Elizabeth (Committee member) / Collins, James (Committee member) / Cooper, Katelyn (Committee member) / Sterner, Beckett (Committee member) / Arizona State University (Publisher)
Created2023
Description
Scientists are entrusted with developing novel molecular strategies for effective prophylactic and therapeutic interventions. Antivirals are indispensable tools that can be targeted at viral domains directly or at cellular domains indirectly to obstruct viral infections and reduce pathogenicity. Despite their transformative potential in healthcare, to date, antivirals have been clinically approved to treat only 10 out of the greater than 200 known pathogenic human viruses. Additionally, as obligate intracellular parasites, many virus functions are intimately coupled with host cellular processes. As such, the development of a clinically relevant antiviral is challenged by the limited number of clear targets per virus and necessitates an extensive insight into these molecular processes. Compounding this challenge, many viral pathogens have evolved to evade effective antivirals. Therefore, a means to develop virus- or strain-specific antivirals without detailed insight into each idiosyncratic biochemical mechanism may aid in the development of antivirals against a larger swath of pathogens. Such an approach will tremendously benefit from having the specific molecular recognition of viral species as the lowest barrier. Here, I modify a nanobody (anti-green fluorescent protein) that specifically recognizes non-essential epitopes (glycoprotein M-pHluorin chimera) presented on the extra virion surface of a virus (Pseudorabies virus strain 486). The nanobody switches from having no inhibitory properties (tested up to 50 μM) to ∼3 nM IC50 in in vitro infectivity assays using porcine kidney (PK15) cells. The nanobody modifications use highly reliable bioconjugation to a three-dimensional wireframe deoxyribonucleic acid (DNA) origami scaffold. Mechanistic studies suggest that inhibition is mediated by the DNA origami scaffold bound to the virus particle, which obstructs the internalization of the viruses into cells, and that inhibition is enhanced by avidity resulting from multivalent virus and scaffold interactions. The assembled nanostructures demonstrate negligible cytotoxicity (<10 nM) and sufficient stability, further supporting their therapeutic potential. If translatable to other viral species and epitopes, this approach may open a new strategy that leverages existing infrastructures – monoclonal antibody development, phage display, and in vitro evolution - for rapidly developing novel antivirals in vivo.
ContributorsPradhan, Swechchha (Author) / Hariadi, Rizal (Thesis advisor) / Hogue, Ian (Committee member) / Varsani, Arvind (Committee member) / Chen, Qiang (Committee member) / Arizona State University (Publisher)
Created2022
Description
Monkeypox virus (MPXV) is an orthopoxvirus that causes smallpox-like disease and has up to a 10% mortality rate, depending on the infectious strain. The global eradication of the smallpox virus has led to the decrease in smallpox vaccinations, which has led to a drastic increase in the number of human MPXV cases. MPXV has been named the most important orthopoxvirus to infect humans since the eradication of smallpox and has been the causative agent of the 2022 world-wide MPXV outbreak. Despite being highly pathogenic, MPXV contains a natural truncation at the N-terminus of its E3 homologue. Vaccinia virus (VACV) E3 protein has two domains: an N- terminus Z-form nucleic acid binding domain (Z-BD) and a C-terminus double stranded RNA binding domain (dsRBD). Both domains are required for pathogenesis, interferon (IFN) resistance, and protein kinase R (PKR) inhibition. The N-terminus is required for evasion of Z-DNA binding protein 1 (ZBP1)-dependent necroptosis. ZBP1 binding to Z- form deoxyribonucleic acid/ribonucleic acid (Z-DNA/RNA) leads to activation of receptor-interacting protein kinase 3 (RIPK3) leading to mixed lineage kinase domain- like (MLKL) phosphorylation, aggregation and cell death. This study investigated how different cell lines combat MPXV infection and how MPXV has evolved ways to circumvent the host response. MPXV is shown to inhibit necroptosis in L929 cells by degrading RIPK3 through the viral inducer of RIPK3 degradation (vIRD) and by inhibiting MLKL aggregation. Additionally, the data shows that IFN treatment efficiently inhibits MPXV replication in a ZBP1-, RIPK3-, and MLKL- dependent manner, but independent of necroptosis. Also, the data suggests that an IFN inducer with a pancaspase or proteasome inhibitor could potentially be a beneficial treatment against MPXV infections. Furthermore, it reveals a link between PKR and pathogen-induced necroptosis that has not been previously described.
ContributorsWilliams, Jacqueline (Author) / Jacobs, Bertram (Thesis advisor) / Langland, Jeffrey (Committee member) / Lake, Douglas (Committee member) / Varsani, Arvind (Committee member) / Arizona State University (Publisher)
Created2022
Description
Traditional public health strategies for assessing human behavior, exposure, and activity are considered resource-exhaustive, time-consuming, and expensive, warranting a need for alternative methods to enhance data acquisition and subsequent interventions. This dissertation critically evaluated the use of wastewater-based epidemiology (WBE) as an inclusive and non-invasive tool for conducting near real-time population health assessments. A rigorous literature review was performed to gauge the current landscape of WBE to monitor for biomarkers indicative of diet, as well as exposure to estrogen-mimicking endocrine disrupting (EED) chemicals via route of ingestion. Wastewater-derived measurements of phytoestrogens from August 2017 through July 2019 (n = 156 samples) in a small sewer catchment revealed seasonal patterns, with highest average per capita consumption rates in January through March of each year (2018: 7.0 ± 2.0 mg d-1; 2019: 8.2 ± 2.3 mg d-1) and statistically significant differences (p = 0.01) between fall and winter (3.4 ± 1.2 vs. 6.1 ± 2.9 mg d-1; p ≤ 0.01) and spring and summer (5.6 ± 2.1 vs. 3.4 ± 1.5 mg d-1; p ≤ 0.01). Additional investigations, including a human gut microbial composition analysis of community wastewater, were performed to support a methodological framework for future implementation of WBE to assess population-level dietary behavior. In response to the COVID-19 global pandemic, a high-frequency, high-resolution sample collection approach with public data sharing was implemented throughout the City of Tempe, Arizona, and analyzed for SARS-CoV-2 (E gene) from April 2020 through March 2021 (n = 1,556 samples). Results indicate early warning capability during the first wave (June 2020) compared to newly reported clinical cases (8.5 ± 2.1 days), later transitioning to a slight lagging indicator in December/January 2020-21 (-2.0 ± 1.4 days). A viral hotspot from within a larger catchment area was detected, prompting targeted interventions to successfully mitigate community spread; reinforcing the importance of sample collection within the sewer infrastructure. I conclude that by working in tandem with traditional approaches, WBE can enlighten a comprehensive understanding of population health, with methods and strategies implemented in this work recommended for future expansion to produce timely, actionable data in support of public health.
ContributorsBowes, Devin Ashley (Author) / Halden, Rolf U (Thesis advisor) / Krajmalnik-Brown, Rosa (Thesis advisor) / Conroy-Ben, Otakuye (Committee member) / Varsani, Arvind (Committee member) / Whisner, Corrie (Committee member) / Arizona State University (Publisher)
Created2022