Matching Items (99)
Description
For cold chain tracking systems, precision and versatility across varying time intervals and temperature ranges remain integral to effective application in clinical, commercial, and academic settings. Therefore, while electronic and chemistry/physics based cold chain tracking mechanisms currently exist, both have limitations that affect their application across various biospecimens and commercial products, providing the initiative to develop a time temperature visual indicator system that resolves challenges with current cold chain tracking approaches. As a result, a permanganate/oxalic acid time temperature visual indicator system for cold chain tracking has been proposed. At thawing temperatures, the designed permanganate/oxalic acid reaction system undergoes a pink to colorless transition as permanganate, Mn(VII), is reduced to auto-catalytic Mn(II), while oxalate is oxidized to CO2. Therefore, when properly stored and vitrified or frozen, the proposed visual indicator remains pink, whereas exposure to thawing conditions will result in an eventual, time temperature dependent, designed color transition that characterizes compromised biospecimen integrity. To design visual indicator systems for targeted times at specific temperatures, absorbance spectroscopy was utilized to monitor permanganate kinetic curves by absorbance at 525 nm. As a result, throughout the outlined research, the following aims were demonstrated: (i) Design and functionality of 1x (0.5 mM KMnO4) visual indicator systems across various time intervals at temperatures ranging from 25°C to -20°C, (ii) Design and functionality of high concentration, 5x, visual indicator systems across varying targeted time intervals at temperatures ranging from 25°C to 0°C, (iii) Pre-activation stability and long-term stability of the proposed visual indicator systems.
ContributorsLjungberg, Emil (Author) / Borges, Chad (Thesis advisor) / Levitus, Marcia (Committee member) / Williams, Peter (Committee member) / Arizona State University (Publisher)
Created2024
Description
Wild horses have roamed the Salt River in Mesa, Arizona since the early 1800s and contribute to the great diversity of the region. Conservation of the herd has been a primary focus for many years and a current focus is population stabilization, but little is known about their virome. Circoviridae, Genomoviridae, and Smacoviridae are the three Cressdnaviricota viruses that have been identified in horses to date. Smacoviridae is classified by the rolling circle replication-associated proteins (Rep) and has a small (2.3-2.9kb), circular, single-stranded genome. The goal of this study was to identify DNA viruses within the fecal samples of the Salt River horses. Samples were collected along the lower Salt River and analyzed in the lab using a metagenomics approach. There were 422 full novel genomes of smacoviruses detected across all samples that were grouped into 144 species based on the similarity of the pairwise identity. Phylogenetic analysis shows the smacoviruses from this study fall into 3 classified genera and the rest cluster into 11 new clades. These results expand the viral diversity associated with wild horses and Smacoviridae, and further studies are needed to determine the host of these viruses.
ContributorsMcGraw, Hannah (Author) / Varsani, Arvind (Thesis director) / Murphree, Julie (Committee member) / Kraberger, Simona (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2024-05
Description
Scorpions are predatory arachnids that are among the most ancient terrestrial invertebrates. They are typically found residing in desert and riparian environments. Viruses associated with scorpions have been explored in the past, unveiling partial RNA virus sequences and polyomaviruses, but more research in this area is necessary. Cycloviruses are non-enveloped viruses with circular single-stranded DNA genomes (~1.7 to 1.9 kb). Cycloviruses were initially identified in mammals and have now been detected in samples from a wide range of mammalian and insect species. Polyomaviruses are double-stranded DNA viruses (~4 to 7 kb). They are known for causing tumors in the host it infects, and have previously been identified in a diverse array of organisms, including scorpions. The objective for this study was to identify known and novel viruses in scorpions. Using high-throughput sequencing and traditional molecular techniques we determine the genome sequences of cycloviruses and polyomaviruses. Sixteen of the forty-three scorpion samples were positive for eight different species of cycloviruses. According to ICTV guidelines, seven of the eight species were novel cycloviruses which were found in bark scorpions, stripe-tailed scorpions, yellow ground scorpions, and giant hairy scorpions (Centruroides sculpturatus, Paravaejovis spinigerus, Paravaejovis confusus & Hadrurus arizonensis) from Maricopa, Pinal, and Pima county in Arizona, USA. Additionally, one previously known cyclovirus species was recovered in bark scorpions (Centruroides sculpturatus) in Pima county which had previously been documented in guano from a Mexican free-tailed bat in Arizona. There were ten scorpions out of forty-three for which we recovered polyomavirus scorpion samples that grouped into four different polyomavirus species. Polyomaviruses were only identified in bark scorpions (Centruroides sculpturatus) from Maricopa, Pinal, and Pima county. Of the polyomavirus genomes recovered three belong to previously identified scorpion polyomavirus 1 and five to scorpion polyomavirus 3, and two represent two new species named scorpion polyomavirus 4 and scorpion polyomavirus 5. The implications of the discovery of cycloviruses and polyomaviruses from this study contributes to our understanding of viral diversity associated with Scorpions.
ContributorsGomez, Magali (Author) / Neil, Julia (Co-author) / Varsani, Arvind (Thesis director) / Kraberger, Simona (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / School of Life Sciences (Contributor)
Created2024-05
Description
Scorpions are predatory arachnids that are among the most ancient terrestrial invertebrates. They are typically found residing in desert and riparian environments. Viruses associated with scorpions have been explored in the past, unveiling partial RNA virus sequences and polyomaviruses, but more research in this area is necessary. Cycloviruses are non-enveloped viruses with circular single-stranded DNA genomes (~1.7 to 1.9 kb). Cycloviruses were initially identified in mammals and have now been detected in samples from a wide range of mammalian and insect species. Polyomaviruses are double-stranded DNA viruses (~4 to 7 kb). They are known for causing tumors in the host it infects, and have previously been identified in a diverse array of organisms, including scorpions. The objective for this study was to identify known and novel viruses in scorpions. Using high-throughput sequencing and traditional molecular techniques we determine the genome sequences of cycloviruses and polyomaviruses. Sixteen of the forty-three scorpion samples were positive for eight different species of cycloviruses. According to ICTV guidelines, seven of the eight species were novel cycloviruses which were found in bark scorpions, stripe-tailed scorpions, yellow ground scorpions, and giant hairy scorpions (Centruroides sculpturatus, Paravaejovis spinigerus, Paravaejovis confusus & Hadrurus arizonensis) from Maricopa, Pinal, and Pima county in Arizona, USA. Additionally, one previously known cyclovirus species was recovered in bark scorpions (Centruroides sculpturatus) in Pima county which had previously been documented in guano from a Mexican free-tailed bat in Arizona. There were ten scorpions out of forty-three for which we recovered polyomavirus scorpion samples that grouped into four different polyomavirus species. Polyomaviruses were only identified in bark scorpions (Centruroides sculpturatus) from Maricopa, Pinal, and Pima county. Of the polyomavirus genomes recovered three belong to previously identified scorpion polyomavirus 1 and five to scorpion polyomavirus 3, and two represent two new species named scorpion polyomavirus 4 and scorpion polyomavirus 5. The implications of the discovery of cycloviruses and polyomaviruses from this study contributes to our understanding of viral diversity associated with Scorpions.
ContributorsNeil, Julia (Author) / Gomez, Magali (Co-author) / Varsani, Arvind (Thesis director) / Kraberger, Simona (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / School of Politics and Global Studies (Contributor)
Created2024-05
Description
Insulator-based dielectrophoresis (iDEP) has attracted considerable attention due to its ability to precisely capture and manipulate nanoparticles and biomolecules. A distinctive approach for effective manipulation of nanometer-sized proteins employing iDEP technique by generating higher electric field (E) and gradient (??2) in the iDEP microfluidic devices is delineated. Strategies to generate higher ??2 in the iDEP devices were outlined using numerical simulations. Intriguingly, the numerical simulation results demonstrated that by decreasing the post-to-post gap in the iDEP microfluidic devices, the ??2 was increased by ⁓12 fold. Furthermore, the inclusion of channel constrictions, such as rectangular constriction or curved constriction into the straight channel iDEP microfluidic device led to a significant increase in ??2. In addition, the inclusion of rectangular constrictions in the straight channel iDEP microfluidic device resulted in a greater increase in ??2 compared to the incorporation of curved constrictions in the same device. Moreover, the straight channel device with horizontal post-to-post gap of 20 μm and vertical post-to-post gap of 10 μm generated the lowest ??2 and the ??2 was uniform across the device. The rectangular constriction device with horizontal and vertical post-to-post gap of 5 μm generated the highest ??2 and the ??2 was non-uniform across the device. Subsequently, suitable candidate devices were fabricated using soft lithography as well as high resolution 3D printing and the DEP behavior of ferritin examined under various experimental conditions. Positive streaming DEP could be observed for ferritin at low frequency in the device generating the lowest ??2, whereas at higher frequency of 10 kHz no DEP trapping characteristics were apparent in the same device. Importantly, in the device geometry resulting in the highest ??2 at 10 kHz, labeled ferritin exhibited pDEPtrapping characteristics. This is an indication that the DEP force superseded diffusion and became the dominant force.
ContributorsMAHMUD, SAMIRA (Author) / Ros, Alexandra (Thesis advisor) / Borges, Chad (Committee member) / Mills, Jeremy (Committee member) / Arizona State University (Publisher)
Created2024
Description
Background: Eosinophilic esophagitis (EoE) is an increasingly prevalent allergic disease characterized by eosinophilic inflammation and symptoms of esophageal dysfunction. Diagnosis and monitoring require repeated, invasive endoscopic esophageal biopsies to assess levels of eosinophilic inflammation. Recently, the minimally invasive esophageal string test (EST) has been used collect protein in mucosal secretions as a surrogate for tissue biopsies in monitoring disease activity. From the string, assessment of the eosinophil-associated proteins major basic protein-1 (MBP-1) and eotaxin-3 (Eot3) is used to assess disease activity; however, this requires measurement in a reference laboratory, for which the turnaround time for results exceeds the time required for histopathologic assessment of endoscopic biopsies. In addition, MBP-1 and Eot3 are not markers unique to eosinophils. These obstacles can be overcome by targeting eosinophil peroxidase (EPX), an eosinophil-specific protein, using a rapid point-of-care test. Currently, EPX is measured by a labor-intensive enzyme-linked immunosorbent assay (ELISA), but we sought to optimize a rapid point-of-care test to measure EPX in EST segments.
Methods: We extracted protein from residual EST segments and measured EPX levels by ELISA and a lateral flow assay (LFA).
Results: EPX levels measured by LFA strongly correlated with those quantified by ELISA
(rs = 0.90 {95% CI: 0.8283, 0.9466}). The EPX LFA is comparable to ELISA for measuring EPX levels in ESTs.
Conclusions: The EPX LFA can provide a way to rapidly test EPX levels in ESTs in clinical settings and may serve as a valuable tool to facilitate diagnosis and monitoring of EoE.
ContributorsDao, Adelyn (Author) / Lake, Douglas (Thesis director) / Borges, Chad (Committee member) / Wright, Benjamin (Committee member) / Barrett, The Honors College (Contributor) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor)
Created2024-05
Description
Alzheimer’s Disease (AD) is the most common form of dementia affecting the population over the age of 65. AD is characterized clinically by increasing difficulty with memory and language, resulting in a loss of independence. This is due to the presence of two characteristic protein aggregates in the brain: extracellular amyloid plaques and intracellular neurofibrillary tangles (NFTs). Utilizing multiplexed immunofluorescence and dimensional reduction analysis the types of cells present in the hippocampus, the region of the brain most affected by AD, can be explored. Understanding the kinds of cell subtypes present, the mechanism behind how AD develops can be explored. Multiplexed IF was performed on human hippocampus FFPE tissues to detect a total of 37 proteins. Dimensional reduction analysis was performed to identify the four major cell types in the brain: neurons, oligodendrocytes, astrocytes, and microglia. After identifying each cell type, further dimensional reduction analysis was performed within each cell type to identify cell subtypes. A total of 21 neuron, 41 oligodendrocyte, 20 astrocyte, and 22 microglia subtypes were identified. The location of cell subtypes in each region of the hippocampal formation was found to match previous reports, further validating the findings of this project.
ContributorsEllison, Mischa A (Author) / Guo, Jia (Thesis advisor) / Borges, Chad (Committee member) / Mastroeni, Diego (Committee member) / Arizona State University (Publisher)
Created2024
Description
Precise modulation of gene expression is essential for proper tissue and cell-specific differentiation and function. Multiple distinct post-transcriptional regulatory mechanisms, such as miRNA (microRNA)-based regulation and alternative polyadenylation (APA), are an intrinsic part of this modulation and orchestrate intricate pathways to achieve and maintain balanced gene expression.MiRNA-based regulation and APA function through sequence motifs located in the 3’ Untranslated Region (3’UTR) of mRNA transcripts. MiRNAs are short (~22 nt) non-coding RNA molecules that bind target sequences within the 3’UTR of an mRNA transcript, inhibiting its translation or promoting its degradation. APA occurs during RNA transcription termination and leads to the preparation of mature mRNAs with different 3’UTR lengths, allowing shorter 3’UTRs to bypass miRNA regulation.
In addition to these two post-transcriptional forms of regulation, co-transcriptional mechanisms such as alternative RNA splicing, which produces distinct gene products from a precursor mRNA, are also important in controlling gene expression.
While miRNA-based regulation, APA, and alternative RNA splicing are important regulatory mechanisms, there is a lack of comprehensive understanding of how they interact and communicate with each other. This thesis studies these three forms of gene regulation in the nematode C. elegans, with the goal of extracting rules and mechanisms used by each of them in development to establish and maintain somatic tissue identity.
After isolating miRNA targets in multiple C. elegans somatic tissues, it was found that miRNAs can modulate the abundance of hnRNPs and SR proteins, which are known to control alternative RNA splicing in a dosage-dependent manner.To identify tissue-specific miRNAs, a nuclear fluorescent cell sorting (FACS)-based methodology named Nuc-Seq, was developed to isolate and sequence tissue-specific miRNAs from body muscle tissue. Nuc-Seq identified 2,848 muscle-specific protein-coding genes and 16 body muscle-specific miRNAs. This data was used to develop a high-quality body muscle-specific miRNA-APA Interactome which allows studies in regulatory processes in detail.
Taken together, this work highlights some of the complexity of pre- and post-transcriptional gene regulation and sheds light on how miRNA-based regulation, APA, and alternative RNA splicing are interconnected and are responsible for the establishment and maintenance of tissue identity.
ContributorsSchorr, Anna L (Author) / Mangone, Marco (Thesis advisor) / Harris, Robin (Committee member) / Sharma, Shalini (Committee member) / Varsani, Arvind (Committee member) / Arizona State University (Publisher)
Created2023
Description
Despite the prevalence of coyotes (Canis latrans) little is known about the viruses associated with this species. To assess the extent of viral research that has been conducted on coyotes, a literature review was performed. Over the last six decades, there have been many viruses that have been identified infecting coyotes. The pathology of some cases implies that infection is rare and lethal while others have been demonstrated to be endemic to coyotes. In addition, the majority of the prior analyses were done through serological assays that were limited to investigating target viruses. To help expand what is known about coyote-virus dynamics, viral assays were conducted on coyote scat. The samples were collected as part of transects established along the Salt River near Phoenix, Arizona, United States (USA). The recovered viral genomes were clustered with other deoxynucleic acid (DNA) viruses and analyzed to determine phylogeny and genetic identity. From the recovered viral genomes, there are two novel circoviruses, one novel naryavirus, five unclassified cressdnaviruses, and two previously identified species of anelloviruses from the Wawtorquevirus genus. For these viruses, new phylogenies for their groups and pairwise identity plots have been generated. These figures give insight into the potential hosts and the evolutionary history. In the case of the anelloviruses, they likely derived from a wood rat (Neotoma) host, given the anellovirus family’s host specificity and its similarity to another viral genome derived from a wood rat in Arizona, USA. Of the recovered circovirus genomes, one is associated with a viral isolate collected from a dust sample in Arizona, USA. The second circovirus species identified is within a clade that consists of rodent associated circoviruses and canine circovirus. Other recovered genomes expand clusters of unclassified cressdnaviruses. The recovered genomes support further genomic analysis. These findings help support the notion that there is a wealth of viral information to be identified from animals like coyotes. By understanding the viruses that coyotes are associated with, it is possible to better understand the viral impact on the urban environment, domesticated animals, and wildlife in general.
ContributorsHess, Savage Cree (Author) / Varsani, Arvind (Thesis advisor) / Kraberger, Simona (Committee member) / Upham, Nathan S (Committee member) / Arizona State University (Publisher)
Created2023
Description
Alpha herpesviruses are a family of neuroinvasive viruses that infect multiplevertebrate species. Alpha herpesviruses are responsible for human and livestock infections, most notably Herpes Simplex Virus (HSV), Varicella Zoster virus (VZV), and Pseudorabies Virus (PRV). PRV is a potent swine virus that can infect other mammals, and results in lethal encephalitis that can be devastating to livestock and of great financial expense to farmers. HSV, types 1 and 2, and VZV are widespread throughout the global human population, with estimates of the HSV-1 burden at about 60% of people worldwide. The hallmark of alpha herpesvirus infection is a persistent, lifelong infection that can reactivate throughout the lifespan of the host. Currently, the precise mechanisms of how these viruses undergo intracellular trafficking to emerge from the infected cell in epithelial tissues is not well understood. Many insights have been made with PRV in animal neurons, both in culture systems and animal models, about the viral genes and host factors involved in these processes. However, understanding of these mechanisms, and the interplay between viral and host proteins, in the human pathogen HSV-1 is even more lacking. Using recombinant fluorescent virus strains of HSV-1 and Total Internal Reflection Microscopy to image the transport of mature viral progeny in epithelial cells, it was determined that the egress of HSV-1 uses constitutive cellular secretory pathways. Specifically, the viral progeny traffic from the trans-Golgi network to the site of exocytosis at the plasma membrane via Rab6a secretory vesicles. This work will contribute to the understanding of how alpha herpesviruses complete their lifecycles in host cells, particularly at the sites where infection initially occurs and can spread to a new organism. Knowledge of these processes may lead to the development of therapeutics or prophylactics to reduce the burden of these viruses.
ContributorsBergeman, Melissa Hope (Author) / Hogue, Ian B (Thesis advisor) / Hogue, Brenda (Committee member) / Roberson, Robert (Committee member) / Varsani, Arvind (Committee member) / Arizona State University (Publisher)
Created2023