Matching Items (82)
Description
Alternative polyadenylation (APA) is the biological mechanism in which the same gene can have multiple 3'untranslated region (3'UTR) isoforms due to the presence of multiple polyadenylation signal (PAS) elements within the pre mRNAs. Because APA produces mRNA transcripts that have different 3'UTR isoforms, certain transcripts may be subject to post-transcriptional regulation by regulatory non-coding RNAs, such as microRNAs or RNA binding proteins defects of which have been implicated in diseases such as cancer. Despite the increasing level of information, functional understanding of the molecular mechanisms involved in transcription is still poorly understood, nor is it clear why APA is necessary at a cell or tissue-specific level. To address these questions I wanted to develop a set of sensor strain plasmids capable of detecting cleavage and polyadenylation in vivo, inject the complete sensor strain plasmid into C. elegans and prepare stable transgenic lines, and perform proof-of-principle RNAi feeding experiments targeting genes associated with the cleavage and polyadenylation complex machinery. I demonstrated that it was possible to create a plasmid capable of detecting cleavage and polyadenylation in C. elegans; however, issues arose during the RNAi assays indicating the sensor strain plasmid was not sensitive enough to the RNAi to effectively detect in the worms. Once the problems involved with sensitivity and variability in the RNAi effects are resolved, the plasmid would be able to better address questions regarding the functional understanding of molecular mechanisms involved in transcription termination.
ContributorsWilky, Henry Patrick (Author) / Mangone, Marco (Thesis director) / Newbern, Jason (Committee member) / Blazie, Stephen (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2015-05
Description
Background
The purpose of this study is to determine the feasibility of three widely used wearable sensors in research settings for 24 h monitoring of sleep, sedentary, and active behaviors in middle-aged women.
Methods
Participants were 21 inactive, overweight (M Body Mass Index (BMI) = 29.27 ± 7.43) women, 30 to 64 years (M = 45.31 ± 9.67). Women were instructed to wear each sensor on the non-dominant hip (ActiGraph GT3X+), wrist (GENEActiv), or upper arm (BodyMedia SenseWear Mini) for 24 h/day and record daily wake and bed times for one week over the course of three consecutive weeks. Women received feedback about their daily physical activity and sleep behaviors. Feasibility (i.e., acceptability and demand) was measured using surveys, interviews, and wear time.
Results
Women felt the GENEActiv (94.7 %) and SenseWear Mini (90.0 %) were easier to wear and preferred the placement (68.4, 80 % respectively) as compared to the ActiGraph (42.9, 47.6 % respectively). Mean wear time on valid days was similar across sensors (ActiGraph: M = 918.8 ± 115.0 min; GENEActiv: M = 949.3 ± 86.6; SenseWear: M = 928.0 ± 101.8) and well above other studies using wake time only protocols. Informational feedback was the biggest motivator, while appearance, comfort, and inconvenience were the biggest barriers to wearing sensors. Wear time was valid on 93.9 % (ActiGraph), 100 % (GENEActiv), and 95.2 % (SenseWear) of eligible days. 61.9, 95.2, and 71.4 % of participants had seven valid days of data for the ActiGraph, GENEActiv, and SenseWear, respectively.
Conclusion
Twenty-four hour monitoring over seven consecutive days is a feasible approach in middle-aged women. Researchers should consider participant acceptability and demand, in addition to validity and reliability, when choosing a wearable sensor. More research is needed across populations and study designs.
The purpose of this study is to determine the feasibility of three widely used wearable sensors in research settings for 24 h monitoring of sleep, sedentary, and active behaviors in middle-aged women.
Methods
Participants were 21 inactive, overweight (M Body Mass Index (BMI) = 29.27 ± 7.43) women, 30 to 64 years (M = 45.31 ± 9.67). Women were instructed to wear each sensor on the non-dominant hip (ActiGraph GT3X+), wrist (GENEActiv), or upper arm (BodyMedia SenseWear Mini) for 24 h/day and record daily wake and bed times for one week over the course of three consecutive weeks. Women received feedback about their daily physical activity and sleep behaviors. Feasibility (i.e., acceptability and demand) was measured using surveys, interviews, and wear time.
Results
Women felt the GENEActiv (94.7 %) and SenseWear Mini (90.0 %) were easier to wear and preferred the placement (68.4, 80 % respectively) as compared to the ActiGraph (42.9, 47.6 % respectively). Mean wear time on valid days was similar across sensors (ActiGraph: M = 918.8 ± 115.0 min; GENEActiv: M = 949.3 ± 86.6; SenseWear: M = 928.0 ± 101.8) and well above other studies using wake time only protocols. Informational feedback was the biggest motivator, while appearance, comfort, and inconvenience were the biggest barriers to wearing sensors. Wear time was valid on 93.9 % (ActiGraph), 100 % (GENEActiv), and 95.2 % (SenseWear) of eligible days. 61.9, 95.2, and 71.4 % of participants had seven valid days of data for the ActiGraph, GENEActiv, and SenseWear, respectively.
Conclusion
Twenty-four hour monitoring over seven consecutive days is a feasible approach in middle-aged women. Researchers should consider participant acceptability and demand, in addition to validity and reliability, when choosing a wearable sensor. More research is needed across populations and study designs.
ContributorsHuberty, Jennifer (Author) / Ehlers, Diane (Author) / Kurka, Jonathan (Author) / Ainsworth, Barbara (Author) / Buman, Matthew (Author) / College of Health Solutions (Contributor) / School of Nutrition and Health Promotion (Contributor)
Created2015-07-30
Description
In eukaryotes, most messenger RNA precursors (pre-mRNA) undergo extensive processing, leading to the cleavage of the transcript followed by the addition of a poly(A) tail. This process is executed by a large complex known as the Cleavage and Polyadenylation Complex (CPC). Its central subcomplex, the Cleavage and Polyadenylation Specificity Factor (CPSF) complex is responsible for recognizing a short hexameric element AAUAAA located at the 3’end in the nascent mRNA molecule and catalyzing the pre-mRNA cleavage. In the round nematode C. elegans, the cleavage reaction is executed by a subunit of this complex named CPSF3, a highly conserved RNA endonuclease. While the crystal structure of its human ortholog CPSF73 has been recently identified, we still do not understand the molecular mechanisms and sequence specificity used by this protein to induce cleavage, which in turn would help to understand how this process is executed in detail. Additionally, we do not understand in additional factors are needed for this process. In order to address these issues, we performed a comparative analysis of the CPSF3 protein in higher eukaryotes to identify conserved functional domains. The overall percent identities for members of the CPSF complex range from 33.68% to 56.49%, suggesting that the human and C. elegans orthologs retain a high level of conservation. CPSF73 is the protein with the overall highest percent identity of the CPSF complex, with its active site-containing domain possessing 74.60% identity with CPSF3. Additionally, we gathered and expressed using a bacterial expression system CPSF3 and a mutant, which is unable to perform the cleavage reaction, and developed an in vitro cleavage assay to test whether CPSF3 activity is necessary and sufficient to induce nascent mRNA cleavage. This project establishes tools to better understand how CPSF3 functions within the CPC and sheds light on the biology surrounding the transcription process as a whole.
ContributorsGallante, Christina (Author) / Mangone, Marco (Thesis director) / Sharma, Shalini (Committee member) / Hrach, Heather (Committee member) / School of Life Sciences (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
The number of cancer survivors in the United States is growing rapidly and it is expected to double by 2040. Arizona is nationally ranked with the 14th highest number of survivors, many of which experience a wide range of persisting medical complications that result from the cancer and associated treatment. Consequently, there is an increased need for services tailored to the health and wellness of survivors. Studies have shown that exercise rehabilitation is effective in improving the physical and mental health of this patient population. This project aimed to investigate the status of medically-based exercise rehabilitation for cancer survivors in Arizona. It focused on services offered by cancer treatment centers and cardiac rehabilitation clinics, with cardiac rehabilitation providing a possible delivery method for future cancer exercise rehabilitation. A directory of resources was compiled based on responses to structured telephone interviews with the cancer treatment centers (n=32) and cardiac rehabilitation clinics (n=34) within the state. The directory will serve as a resource for both patients and clinicians by identifying statewide related services that are available at the medical institutions and within the community. Results showed that 42.9% and 39.4% of the cancer treatment centers and cardiac rehabilitation clinics, respectively, offered exercise related services for cancer survivors. 78.6% of cancer centers stated that they refer cancer survivors to physical therapy, while only 35.7% refer survivors to community-based programs. Only 2 cardiac rehabilitation clinics, or 6%, offered preventative cardiology exercise consultations to cancer survivors. In conclusion, rehabilitative exercise resources for cancer survivors in Arizona were limited. Additional cancer rehabilitation efficacy studies are needed to further clarify evidence-based practice guidelines and provide direction for optimal methods of healthcare delivery. It is recommended that this directory remains current with routine updates in an effort to increase patient accessibility to care.
ContributorsHitt, Ellen (Author) / Scales, Robert (Thesis director) / Huberty, Jennifer (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-12
Description
The successful reduction of CO2 and protons by a light-induced cobalt porphyrin/cytb562 hybrid metalloenzyme in water is reported. Incorporation of the porphyrin into a protein scaffold results in increases in CO and H2 production over naked porphyrin. Rational point mutations to the CoPPIX binding site of cytb562 modulate production, indicating possible further improvements in catalytic activity.
ContributorsGwerder, Noah D (Author) / Ghirlanda, Giovanna (Thesis director) / Williams, Peter (Committee member) / Mangone, Marco (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
Duchenne muscular dystrophy (DMD) is a lethal, X-linked disease which occurs in approximately 1 in 3,500 male births. This disease is characterized by progressive muscle wasting and causes premature death. One of the earliest symptoms of this disease is mitochondrial dysfunction. Dystrophin is a protein found under the sarcolemma. The N terminus binds to actin and the C terminus binds to dystrophin glycoprotein complex (DGC). DMD is caused by mutations in the dystrophin gene. C. elegans possess an ortholog of dystrophin, DYS-1. Though there is evidence that C. elegans can be used as a model organism to model DMD, nematode DGC has not been well characterized. Additionally, while we know that mitochondrial dysfunction has been found in humans and other model organisms, this has not been well defined in C. elegans. In order to address these issues, we crossed the SJ4103 worm strain (myo-3p::GFP(mit)) with dys-1(cx18) in order to visualize and quantify changes in mitochondria in a dys-1 background. SJ4103;cx18 nematodes were found to have less mitochondrial than SJ4103 which suggests mitochondrial dysfunction does occur in dys-1 worms. Furthermore, mitochondrial dysfunction was studied by knocking down members of the DGC, dys-1, dyb-1, sgn-1, sgca-1, and sgcb-1 in SJ4103 strain. Knock down of each gene resulted in decrease in abundance of mitochondria which suggests that each member of the DGC contributes to the overall health of nematode muscle. The ORF of dyb-1 was successfully cloned and tagged with GFP in order to visualize this DGC member C. elegans. Imaging of the transgenic dyb-1::GFP worm shows green fluoresce expressed in which suggests that dyb-1 is a functional component of the muscle fibers. This project will enable us to better understand the effects of dystrophin deficiency on mitochondrial function as well as visualize the expression of certain members of the DGC in order to establish C. elegans as a good model organism to study this disease.
ContributorsObrien, Shannon Nishino (Author) / Mangone, Marco (Thesis director) / Newbern, Jason (Committee member) / Hrach, Heather (Committee member) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Cleavage and polyadenylation is a step in mRNA processing in which the 3’UTR is cleaved and a polyA tail is added to create a final mature transcript. This process relies on RNA sequence elements that guide a large multimeric protein complex named the Cleavage and Polyadenylation Complex to dock on the 3’UTR and execute the cleavage reaction. Interactions of the complex with the RNA and specific dynamics of complex recruitment and formation still remain largely uncharacterized. In our lab we have identified an Adenosine residue as the nucleotide most often present at the cleavage site, although it is unclear whether this specific element is a required instructor of cleavage and polyadenylation. To address whether the Adenosine residue is necessary and sufficient for the cleavage and polyadenylation reaction, we mutated this nucleotide at the cleavage site in three C. elegans protein coding genes, forcing the expression of these wt and mutant 3’UTRs, and studied how the cleavage and polyadenylation machinery process these genes in vivo. We found that interrupting the wt sequence elements found at the cleavage site interferes with the cleavage and polyadenylation reaction, suggesting that the sequence close to the end of the transcript plays a role in modulating the site of the RNA cleavage. This activity is also gene-specific. Genes such as ges-1 showed little disruption in the cleavage of the transcript, with similar location occurring in both the wt and mutant 3’UTRs. On the other hand, mutation of the cleavage site in genes such as Y106G6H.9 caused the activation of new cryptic cleavage sites within the transcript. Taken together, my experiments suggest that the sequence elements at the cleavage site somehow participate in the reaction to guide the cleavage reaction to occur at an exact site. This work will help to better understand the mechanisms of transcription termination in vivo and will push forward research aimed to study post-transcriptional gene regulation in eukaryotes.
ContributorsSteber, Hannah Suzanne (Author) / Mangone, Marco (Thesis director) / Harris, Robin (Committee member) / LaBaer, Joshua (Committee member) / School of Life Sciences (Contributor, Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Background:
The positive impacts of yoga on stress, pain, and chronic disease has recently led to the integration of yoga as part of physical therapy (PT) treatment. Due to the lack of training for PTs related to yoga, there is currently a need to provide knowledge and education about how to safely and easily implement therapeutic yoga (TY) as a complementary treatment approach.
Objective:
The purpose of this study was to assess the readiness of PTs (those who do not currently prescribe TY to patients) to integrate TY into treatment, and secondly, the feasibility (i.e., acceptability, demand, and practicality) of a 5-week online TY training to improve the readiness of PT’s to utilize TY in their practice.
Methods:
Licensed Physical Therapist’s (n=103) were recruited nationally through social media and email. Eligible and consented participants were asked to register in a 5-week online TY training course, Readiness for Integrating Yoga Therapeutics into Rehabilitation for PTs (intervention). PTs perceptions of TY and the role of safety and confidence in prescribing TY to patients were measured at baseline and post-intervention using a customized survey. Feasibility outcomes were measured after completion of the 5-week online training course with a survey. Feasibility was measured with acceptability, demand, and practicality. Our benchmarks included: (1) at least 70% of PTs would find the course acceptable, (2) at least 60% would finish the course (i.e., demand) and (3) there would be significant improvements in PTs perceptions of TY.
Results:
A total of 95 licensed PTs registered in the 5-week online TY training course, with 60 PTs (63%) completing the intervention and surveys. Of the PTs who completed the 5-week online training course, most PTs felt they were not ready (n=19/60, 31.7%) or somewhat ready (n=25/60, 41.7%) to integrate TY prior to taking the online training. Over half of PTs thought the online training was acceptable (n= 50/60, 83.3%) and finished the course (n=60/95, 63%). There were significant improvements in personal readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients.
Conclusion:
Findings suggest a 5-week online TY training course is feasible in improving PTs readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients. Future studies are proposed to test the effectiveness of TY training and education opportunities with PTs to further advance the adoption of TY into PT practice.
The positive impacts of yoga on stress, pain, and chronic disease has recently led to the integration of yoga as part of physical therapy (PT) treatment. Due to the lack of training for PTs related to yoga, there is currently a need to provide knowledge and education about how to safely and easily implement therapeutic yoga (TY) as a complementary treatment approach.
Objective:
The purpose of this study was to assess the readiness of PTs (those who do not currently prescribe TY to patients) to integrate TY into treatment, and secondly, the feasibility (i.e., acceptability, demand, and practicality) of a 5-week online TY training to improve the readiness of PT’s to utilize TY in their practice.
Methods:
Licensed Physical Therapist’s (n=103) were recruited nationally through social media and email. Eligible and consented participants were asked to register in a 5-week online TY training course, Readiness for Integrating Yoga Therapeutics into Rehabilitation for PTs (intervention). PTs perceptions of TY and the role of safety and confidence in prescribing TY to patients were measured at baseline and post-intervention using a customized survey. Feasibility outcomes were measured after completion of the 5-week online training course with a survey. Feasibility was measured with acceptability, demand, and practicality. Our benchmarks included: (1) at least 70% of PTs would find the course acceptable, (2) at least 60% would finish the course (i.e., demand) and (3) there would be significant improvements in PTs perceptions of TY.
Results:
A total of 95 licensed PTs registered in the 5-week online TY training course, with 60 PTs (63%) completing the intervention and surveys. Of the PTs who completed the 5-week online training course, most PTs felt they were not ready (n=19/60, 31.7%) or somewhat ready (n=25/60, 41.7%) to integrate TY prior to taking the online training. Over half of PTs thought the online training was acceptable (n= 50/60, 83.3%) and finished the course (n=60/95, 63%). There were significant improvements in personal readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients.
Conclusion:
Findings suggest a 5-week online TY training course is feasible in improving PTs readiness to prescribe TY, safety prescribing TY, confidence to prescribe TY, current understanding/knowledge of TY and feeling adequately trained and educated to use some form of TY techniques with patients. Future studies are proposed to test the effectiveness of TY training and education opportunities with PTs to further advance the adoption of TY into PT practice.
ContributorsThompson, Abigail Ann (Co-author) / Thompson, Abigail (Co-author) / Huberty, Jennifer (Thesis director) / Taylor, Matthew (Committee member) / Ortiz, Alexis (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Abstract Objective: The purpose of this study was to determine the feasibility (e.g., practicality and demand) of a 4-week series of yoga classes in a homeless shelter. Participants: Five current residents of Central Arizona Shelter Services (CASS) and the Chief of Programming at CASS. Methods: Each shelter resident participated in a 5-minute interview answering questions regarding the demand of implementing a yoga program at CASS. The Chief of Programming participated in a 30-minute interview answering questions regarding the practicality of implementing a 4-week series yoga program at the homeless shelter. Results: CASS residents reported a strong desire to attend a yoga program. The Chief of Programming at CASS reported that implementing a yoga program would conflict with the overall goal of the shelter. Conclusion: Implementing a 4-week series yoga program is not feasible at CASS although there is a strong demand for a yoga program among the homeless population of the Phoenix metro area.
ContributorsSamuels, Jasmyne Angelique (Author) / Hart, Teresa (Thesis director) / Huberty, Jennifer (Committee member) / Pearl, Julia (Committee member) / School of Nutrition and Health Promotion (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
Description
Recent research has confirmed and revealed many physical and mental benefits of yoga. The practice of yoga has spread throughout the western world, where it is widely used for the purpose of exercise and fitness. Due to its rise in popularity, there is a need for research regarding the energy expenditure required for different types of yoga. The majority of the literature cites yoga as being an effective exercise for light intensity activity, but there are not as many studies attempting to determine if there are postures and sequences that can meet the requirements for moderate physical activity. In addition, there is a need to validate mobile devices with which to measure energy expenditure (EE) that are compatible with the dynamic movements that occur during yoga. The purpose of this study was to measure energy expenditure of twenty-two yoga practitioners of varying experience during a 30-minute Vinyasa flow yoga practice and from this data collection determine: if an ashtanga-based vinyasa yoga class meets the criteria for moderate intensity physical activity, the reliability between the Actigraph and Oxycon Mobile and the validity of an Actigraph GT3X device worn on the hip in estimating energy expenditure for ashtanga-based vinyasa flow yoga. The Actigraph GT3X and the Oxycon mobile were used to measure energy expenditure. Mean values for energy expenditure recorded by the Oxycon and Actigraph were 3.19 ± 0.42 METs and 1.16 ± 0.23 METs respectively, exhibiting a significant difference in data collection. There was no correlation between the values recorded by the two devices, indicating that the Actigraph was not consistent with the Oxycon Mobile (previously validated for measurement of EE). Results of this study indicate that this Vinyasa flow yoga sequence does satisfy the criteria for moderate intensity physical activity as defined by ACSM with an average EE of 3.19 ± 0.42 METs, and that the Actigraph GT3X is not an accurate device for measurement of EE for yoga.
ContributorsHand, Lindsay Gabrielle (Author) / Huberty, Jennifer (Thesis director) / Buman, Matthew (Committee member) / School of Nutrition and Health Promotion (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12