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Description
Calcium is the only ion capable of triggering electrical and chemical reactions in cells which are part of essential biomolecular processes, such as gene transcription and ion flux. Calcium homeostasis, the control of concentration levels, is therefore crucial for the proper functioning of cells. For example, cardiomyocytes, the cells that form cardiac muscle, rely on calcium transfer process to produce muscle contraction.
The purpose of this work is to study aspects of calcium homeostasis in the model organism Saccharomyces cerevisiae, common yeast. Using luminometric techniques, the response of the yeast was monitored against a set of changes in the environment calcium abundance. The results indicate a complex response as both increase and decreases of external calcium induce elevations in cytosolic calcium concentrations.
Calcium is transferred across compartments by means of channels. In Saccharomyces cerevisiae, many of them have been identified; Cch1p-Mid1p, Vcx1p, Pmc1p, Pmr1p, and Yvc1p. Their participation in calcium homeostasis is well established. Observations of cytosolic calcium increase after a hypertonic shock are mainly associated with influx of ions from the environment though the Cch1p-Mid1p. This process is generally considered as driven by calcium concentration gradients. However, recent studies have suggested that the plasma membrane channel, Cch1p-Mid1p, may possess more sophisticated regulation and sensory mechanisms. The results of our experiments support these ideas.
We carried out experiments that subjected yeast to multiple shocks: a hypertonic shock followed by either a second hypertonic shock, a hypotonic shock, or a yeast dilution pulse where the solution volume increases by the calcium concentration has only a small change. The cytosolic calcium concentration of a yeast population was monitored via luminometry.
The main result of this study is the observation of an unexpected response to the combination of hypertonic and hypotonic shocks. In this case it was observed that the cytosolic calcium concentration increased after both shocks. This indicates that cytosolic calcium increases are not solely driven by the presence of concentration gradients. The response after the hypotonic pulse arises from more complex mechanisms that may include sensor activity at the membrane channels and the release of calcium from internal storages.
The purpose of this work is to study aspects of calcium homeostasis in the model organism Saccharomyces cerevisiae, common yeast. Using luminometric techniques, the response of the yeast was monitored against a set of changes in the environment calcium abundance. The results indicate a complex response as both increase and decreases of external calcium induce elevations in cytosolic calcium concentrations.
Calcium is transferred across compartments by means of channels. In Saccharomyces cerevisiae, many of them have been identified; Cch1p-Mid1p, Vcx1p, Pmc1p, Pmr1p, and Yvc1p. Their participation in calcium homeostasis is well established. Observations of cytosolic calcium increase after a hypertonic shock are mainly associated with influx of ions from the environment though the Cch1p-Mid1p. This process is generally considered as driven by calcium concentration gradients. However, recent studies have suggested that the plasma membrane channel, Cch1p-Mid1p, may possess more sophisticated regulation and sensory mechanisms. The results of our experiments support these ideas.
We carried out experiments that subjected yeast to multiple shocks: a hypertonic shock followed by either a second hypertonic shock, a hypotonic shock, or a yeast dilution pulse where the solution volume increases by the calcium concentration has only a small change. The cytosolic calcium concentration of a yeast population was monitored via luminometry.
The main result of this study is the observation of an unexpected response to the combination of hypertonic and hypotonic shocks. In this case it was observed that the cytosolic calcium concentration increased after both shocks. This indicates that cytosolic calcium increases are not solely driven by the presence of concentration gradients. The response after the hypotonic pulse arises from more complex mechanisms that may include sensor activity at the membrane channels and the release of calcium from internal storages.
ContributorsMintz, David Anthony (Co-author) / Parker, Augustus (Co-author) / Solis, Francisco (Thesis director) / Marshall, Pamela (Committee member) / School of Mathematical and Natural Sciences (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
Calcium is the only ion capable of triggering electrical and chemical reactions in cells which are part of essential biomolecular processes, such as gene transcription and ion flux. Calcium homeostasis, the control of concentration levels, is therefore crucial for the proper functioning of cells. For example, cardiomyocytes, the cells that form cardiac muscle, rely on calcium transfer process to produce muscle contraction.
The purpose of this work is to study aspects of calcium homeostasis in the model organism Saccharomyces cerevisiae, common yeast. Using luminometric techniques, the response of the yeast was monitored against a set of changes in the environment calcium abundance. The results indicate a complex response as both increase and decreases of external calcium induce elevations in cytosolic calcium concentrations.
Calcium is transferred across compartments by means of channels. In Saccharomyces cerevisiae, many of them have been identified; Cch1p-Mid1p, Vcx1p, Pmc1p, Pmr1p, and Yvc1p. Their participation in calcium homeostasis is well established. Observations of cytosolic calcium increase after a hypertonic shock are mainly associated with influx of ions from the environment though the Cch1p-Mid1p. This process is generally considered as driven by calcium concentration gradients. However, recent studies have suggested that the plasma membrane channel, Cch1p-Mid1p, may possess more sophisticated regulation and sensory mechanisms. The results of our experiments support these ideas.
We carried out experiments that subjected yeast to multiple shocks: a hypertonic shock followed by either a second hypertonic shock, a hypotonic shock, or a yeast dilution pulse where the solution volume increases by the calcium concentration has only a small change. The cytosolic calcium concentration of a yeast population was monitored via luminometry.
The main result of this study is the observation of an unexpected response to the combination of hypertonic and hypotonic shocks. In this case it was observed that the cytosolic calcium concentration increased after both shocks. This indicates that cytosolic calcium increases are not solely driven by the presence of concentration gradients. The response after the hypotonic pulse arises from more complex mechanisms that may include sensor activity at the membrane channels and the release of calcium from internal storages.
The purpose of this work is to study aspects of calcium homeostasis in the model organism Saccharomyces cerevisiae, common yeast. Using luminometric techniques, the response of the yeast was monitored against a set of changes in the environment calcium abundance. The results indicate a complex response as both increase and decreases of external calcium induce elevations in cytosolic calcium concentrations.
Calcium is transferred across compartments by means of channels. In Saccharomyces cerevisiae, many of them have been identified; Cch1p-Mid1p, Vcx1p, Pmc1p, Pmr1p, and Yvc1p. Their participation in calcium homeostasis is well established. Observations of cytosolic calcium increase after a hypertonic shock are mainly associated with influx of ions from the environment though the Cch1p-Mid1p. This process is generally considered as driven by calcium concentration gradients. However, recent studies have suggested that the plasma membrane channel, Cch1p-Mid1p, may possess more sophisticated regulation and sensory mechanisms. The results of our experiments support these ideas.
We carried out experiments that subjected yeast to multiple shocks: a hypertonic shock followed by either a second hypertonic shock, a hypotonic shock, or a yeast dilution pulse where the solution volume increases by the calcium concentration has only a small change. The cytosolic calcium concentration of a yeast population was monitored via luminometry.
The main result of this study is the observation of an unexpected response to the combination of hypertonic and hypotonic shocks. In this case it was observed that the cytosolic calcium concentration increased after both shocks. This indicates that cytosolic calcium increases are not solely driven by the presence of concentration gradients. The response after the hypotonic pulse arises from more complex mechanisms that may include sensor activity at the membrane channels and the release of calcium from internal storages.
ContributorsParker, Augustus Carrucciu (Co-author) / Mintz, David (Co-author) / Solis, Francisco (Thesis director) / Marshall, Pamela (Committee member) / School of Mathematical and Natural Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
High childhood obesity rates have resulted in many interventions to attempt to lower these rates. Interventions such as day camps, residential camps, therapy-based interventions and family-based interventions lead to changes in weight and self-esteem but family-based intervention leads to the longest-term success for children ages nine to 17. Analysis of the interventions was measured using tools such as BMI, BMI-percentiles, and weight. Psychological measures such as self-esteem, happiness, and quality of life analysis was preferred, however were not measured in all studies. While most interventions resulted in weight loss and increased self-esteem, results were often not long-term. Studies provided evidence that family-based therapy has potential to last long-term, however there is a lack of research. To determine the most effective childhood nutrition intervention research must conduct follow-ups for many years after the initial intervention to ensure they provide long-term results.
ContributorsAnderson, Megan Lee (Author) / McCoy, Maureen (Thesis director) / Kniskern, Megan (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
This project details the synthesis and analysis of five analogs of model compound NEt-4IB (6-[ethyl(4-isobutoxy-3-isopropylphenyl)amino]nicotinic acid), that target the retinoid-X-receptor (RXR). These molecules were synthesized by substituting, adding, or removing substituents in the nitrogen-containing ring of NEt-4IB. The parent compound is a RXR partial agonist and has proven to be effective in the treatment of type II diabetes without the unwanted side effects seen with full agonists. Many of the current drugs used to treat type II diabetes are accompanied by adverse effects including increased triglyceride levels, weight gain, and hypoglycemia. Biological evaluation with KK-Ay (obese diabetic) model mice indicates that NEt-4IB may even be more effective than current drugs on the market, like pioglitazone. As a result, it is predicted that due to such structural similarity, the analogs synthesized for this work will perform equally, if not better than, NEt-4IB.
ContributorsMaiorella, Emma Lauren (Author) / Wagner, Carl (Thesis director) / Marshall, Pamela (Committee member) / School of Mathematical and Natural Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
In humans, infections, disease, inflammation, and other injuries to specific tissues have been shown to cause delays in the onset of puberty. It is known that steroid hormones and insulin play a role in these delays, yet it is not understood what is happening with the immune system during this response. Similar results have been found in the fruit fly, Drosophila melanogaster, in which damage to adult precursor tissues triggers developmental delays. This project addresses the immune component of the injury response in Drosophila. The goal is to identify which immune response genes, if any, show a significant change in expression after injury. The general methodologies used were first inducing injury via a temperature- sensitive expression of cell death genes in wing precursor tissues, then examining changes in gene expression of immune response genes before and after injury using real-time PCR. The results show that injury increases the expression of genes Drs, CecA1, and Def while decreasing expression of Rel, Dpt, PGRP-LE, and Tl. The changes in immune gene expression following injury suggest the possibility of an immune component to the systemic injury response. These results can further be explored by using mutations of the immune genes to examine their direct effects on the systemic injury response. This research can eventually lead to preventative measures to protect against developmental delays due to infections and diseases in humans.
ContributorsDuprey, Deanna Jeanette (Author) / Hackney, Jennifer (Thesis director) / Marshall, Pamela (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Natural Sciences (Contributor) / School of Social and Behavioral Sciences (Contributor)
Created2014-12
Description
Virtually all animals require relatively predictable developmental schedules in order to fulfill the cycle of life. Cell death and severe inflammation alter steroid hormone production and can disrupt the timing of developmental transitions such as puberty. In the fruit fly, Drosophila melanogaster, injury to wing precursor tissues has been shown to result in decreased steroid hormone levels and delay development. The effects of damage to other tissues have not yet been explored. Here, the larval salivary glands were damaged in order to observe how injuring these specific tissues affect the timing of developmental transitions. Damage was induced by tissue-specific, temperature sensitive activation of cell death genes. The results indicated that death to salivary gland cells accelerates the Drosophila time to adult eclosion and that the observed acceleration of development is age-dependent. Insight into the effects of injury on development in Drosophila can potentially lead to information about development in other organisms, including humans, following injury or chronic inflammation.
ContributorsRippere, Alicia Leann (Author) / Hackney, Jennifer (Thesis director) / Marshall, Pamela (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Natural Sciences (Contributor)
Created2015-05
Description
Bexarotene (Targretin®) is an FDA approved drug used to treat cutaneous T-cell lymphoma (CTCL), as well as off-label treatments for various cancers and neurodegenerative diseases. Previous research has indicated that bexarotene has a specific affinity for retinoid X receptors (RXR), which allows bexarotene to act as a ligand-activated-transcription factor and in return control cell differentiation and proliferation. Bexarotene targets RXR homodimerization to drive transcription of tumor suppressing genes; however, adverse reactions occur simultaneously when bound to other nuclear receptors. In this study, we used novel bexarotene analogs throughout 5 iterations synthesized in the laboratory of Dr. Wagner to test for their potency and ability to bind RXR. The aim of our study is to quantitatively measure RXR homodimerization driven by bexarotene analogs using a yeast two-hybrid system. Our results suggests there to be several compounds with higher protein activity than bexarotene, particularly in generations 3.0 and 5.0. This higher affinity for RXR homodimers may help scientists identify a compound that will minimize adverse effects and toxicity of bexarotene and serve as a better cancer treatment alternative.
ContributorsSeto, David Hua (Author) / Marshall, Pamela (Thesis director) / Wagner, Carl (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Natural Sciences (Contributor) / School of Social and Behavioral Sciences (Contributor)
Created2015-05
Description
The purpose of this study was to develop proposal lesson plans for 4th-6th graders based on active learning to integrate movement physical activity into the curriculum. The 4th-6th graders were chosen, as this is the age where teaching typically transitions from active learning to sedentary/lecture style teaching. Research compiled indicated positive effects of active based learning on children such as increased attention span, retention, and general focus. A survey was created to not only assess the perception of active versus didactic learners, but to also assess the effects of movement-based learning on the variables that research claimed to change. The lesson plans developed here should be transferable to a classroom lesson to evaluate the hypothesized results.
ContributorsTanna, Nimisha (Author) / Hyatt, JP (Thesis director) / Ainsworth, Barbara (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
About 75% of men and 66.58% of women are considered overweight or obese (BMI ≥25). $117 billion dollars is spent each year in medical costs due to physical inactivity. Aerobic exercise has been well defined in its’ benefits to cardiovascular health; however, the effects of resistance training are still not well defined. The purpose of this preliminary analysis was to evaluate the vascular health effects (central and peripheral blood pressure and VO2 max) of two different types of resistance training programs: high load, low repetitions resistance training and low load, high repetitions resistance training. Fourteen participants aged 18-55 years (6 males, 8 females) were involved in this preliminary analysis. Data were collected before and after the 12-week long exercise program (36 training sessions) via pulse wave analysis and VO2peak testing. Multivariate regression analysis of training program effects, while adjusting for body mass index and time, did not result in significant training effects on central and peripheral diastolic blood pressure, nor VO2peak. A statistical trend was observed between the different training programs for systolic blood pressure, suggesting that subjects partaking in the high load, low repetitions program exhibited higher systolic blood pressures than the low load, high repetitions group. With a larger sample size, the difference in systolic blood pressure may increase between training program groups and indicate that greater loads with minimal repetitions may increase lead to clinically significant elevations in blood pressure. Further work is needed to uncover the relationship between different types of resistance training and blood pressure, especially if these lifting regimens are continued for longer lengths of time.
ContributorsHill, Cody Alan (Co-author) / Hill, Cody (Co-author) / Whisner, Corrie (Thesis director) / Angadi, Siddhartha (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Introduction: Individuals with rotator cuff tears have been found to compensate in their movement patterns by using lower thoracohumeral elevation angles during certain tasks, as well as increased internal rotation of the shoulder (Vidt et al., 2016). Leading joint hypothesis suggests there is one leading joint that creates the foundation for the entire limb motion, and there are other subordinate joints which monitor the passive interaction torque and create a net torque aiding to limb motions required for the task. This experiment seeks to establish a better understanding of joint control strategies during a wide range of arm movements. Based on the leading joint hypothesis, we hypothesize that when a subject has a rotator cuff tear, their performance of planar and three-dimensional motions should be altered not only at the shoulder, which is often the leading joint, but also at other joints on the arm, such as the elbow and wrist. This paper will focus on the effect of normal aging on the control of the joints of the arm.
Methods: There were 4 groups of participants: healthy younger adults (n=14)(21.74 ± 1.97), healthy older adults (n=12)(55-75), older adults (n=4)(55-75) with a partial-thickness rotator cuff tear, and older adults (n=4)(55-75) with a full-thickness rotator cuff tear (RCT). All four groups completed strength testing, horizontal drawing and pointing tasks, and three dimensional (3D) activities of daily living. Kinematic and kinetic variables of the arm were obtained during horizontal and 3D tasks using data from 12 reflective markers placed on the arm, 8 motion capture cameras, and Cortex motion capture software (Motion Analysis Corp., Santa Rosa, CA). Strength testing tasks were measured using a dynamometer. All strength testing and 3D tasks were completed for three trials and horizontal tasks were completed for two trials.
Results: Results of the younger adult participants showed that during the forward portion of seven 3D tasks, there were four phases of different joint control mechanics seen in a majority of the movements. These phases included active rotation of both the shoulder and the elbow joint, active rotation of the shoulder with passive rotation of the elbow, passive rotation of the shoulder with active rotation of the elbow, and passive rotation of both the shoulder and the elbow. Passive rotation during movements was a result of gravitational torque on the different segments of the arm and interaction torque caused as a result of the multi-joint structure of human limbs. The number of tested participants for the minor RCT, and RCT older adults groups is not yet high enough to produce significant results and because of this their results are not reported in this article. Between the older adult control group and the young adult control group in the tasks upward reach to eye height and hair comb there were significant differences found between the groups. The differences were found in shorter overall time and distance between the two groups in the upward eye task.
Discussion: Through the available results, multiple phases were found where one or both of the joints of the arm moved passively which further supports the LJH and extends it to include 3D movements. With available data, it can be concluded that healthy older adults use movement control strategies, such as shortening distance covered, decreasing time percentage in active joint phases, and increasing time percentage in passive joint phases, to account for atrophy along with other age-related declines in performance, such as a decrease in range of motion. This article is a part of a bigger project which aims to better understand how older adults with RCTs compensate for the decreased strength, the decreased range of motion, and the pain that accompany this type of injury. It is anticipated that the results of this experiment will lead to more research toward better understanding how to treat patients with RCTs.
Methods: There were 4 groups of participants: healthy younger adults (n=14)(21.74 ± 1.97), healthy older adults (n=12)(55-75), older adults (n=4)(55-75) with a partial-thickness rotator cuff tear, and older adults (n=4)(55-75) with a full-thickness rotator cuff tear (RCT). All four groups completed strength testing, horizontal drawing and pointing tasks, and three dimensional (3D) activities of daily living. Kinematic and kinetic variables of the arm were obtained during horizontal and 3D tasks using data from 12 reflective markers placed on the arm, 8 motion capture cameras, and Cortex motion capture software (Motion Analysis Corp., Santa Rosa, CA). Strength testing tasks were measured using a dynamometer. All strength testing and 3D tasks were completed for three trials and horizontal tasks were completed for two trials.
Results: Results of the younger adult participants showed that during the forward portion of seven 3D tasks, there were four phases of different joint control mechanics seen in a majority of the movements. These phases included active rotation of both the shoulder and the elbow joint, active rotation of the shoulder with passive rotation of the elbow, passive rotation of the shoulder with active rotation of the elbow, and passive rotation of both the shoulder and the elbow. Passive rotation during movements was a result of gravitational torque on the different segments of the arm and interaction torque caused as a result of the multi-joint structure of human limbs. The number of tested participants for the minor RCT, and RCT older adults groups is not yet high enough to produce significant results and because of this their results are not reported in this article. Between the older adult control group and the young adult control group in the tasks upward reach to eye height and hair comb there were significant differences found between the groups. The differences were found in shorter overall time and distance between the two groups in the upward eye task.
Discussion: Through the available results, multiple phases were found where one or both of the joints of the arm moved passively which further supports the LJH and extends it to include 3D movements. With available data, it can be concluded that healthy older adults use movement control strategies, such as shortening distance covered, decreasing time percentage in active joint phases, and increasing time percentage in passive joint phases, to account for atrophy along with other age-related declines in performance, such as a decrease in range of motion. This article is a part of a bigger project which aims to better understand how older adults with RCTs compensate for the decreased strength, the decreased range of motion, and the pain that accompany this type of injury. It is anticipated that the results of this experiment will lead to more research toward better understanding how to treat patients with RCTs.
ContributorsFlores, Noah Mateo (Author) / Dounskaia, Natalia (Thesis director) / Vidt, Meghan (Committee member) / College of Health Solutions (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2019-05