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Deconstructing bioluminescence: from molecular detail to in vivo imaging.Adams, Spencer T., Jr. 29 January 2020 (has links)
Bioluminescence is the chemical production of light that results when a luciferase enzyme catalyzes the luminogenic oxidation of a small-molecule luciferin substrate. The numerous luciferases and luciferins nature has evolved can be used to illuminate biological processes, from in vitro assays to imaging processes in live animals. However, we can improve the utility of bioluminescence through modification of these enzymes and substrates. My thesis work focuses on developing reporters that expand the bioluminescent toolkit and improving our understanding of how bioluminescence works on a molecular level.
The first part of my thesis focuses on characterizing luciferases and luciferins that improve bioluminescence imaging in vivo. Some of our luciferins can outperform the natural D-luciferin substrate in live mouse imaging, while others are selectively utilized by mutant luciferases in live mouse brain. We also engineered luciferins that can selectively report on endogenous enzymatic activity in live mice.
The second part of my thesis focuses on determining the molecular details of how enzymes related to firefly luciferase, long-chain fatty acyl-CoA synthetases (ACSLs), can function as latent luciferases. I have determined the structure for one of these enzymes and improved its bioluminescent activity with synthetic luciferins enough to image in live mouse brain. I also characterized the selectivity in chimerized enzymes that combine firefly luciferase and ACSLs. In summary, my work improves the utility of bioluminescence for in vivo use and informs us about how evolutionarily-related enzymes function as luciferases on a molecular level.
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Internalized Shame as a Moderating Variable for Inhibited Sexual Difficulties in Adult Women Resulting From Childhood Sexual AbuseRobinson, Kristine 23 May 2006 (has links) (PDF)
An adult female sample of childhood sexual abuse survivors (N=467) were surveyed to determine whether internalized shame moderated the effects of sexual inhibited difficulties. Other variables such as severity, duration, and frequency of sexual abuse, as well as whether physical abuse was also part of their experience, were examined to determine their role in later sexual inhibition. It was predicted that there would be a significant positive relationship between 1) Scores on variables of physical abuse, severity of abuse, frequency of abuse, duration of abuse, identity of the perpetrator and scores on the variable of inhibited sexual difficulties; 2) Scores of internalized shame and scores of inhibited sexual difficulties and 3) Scores on variable of physical abuse, severity of abuse, frequency of abuse, duration of abuse, identity of the perpetrator and scores on the variables of internalized shame. Through Structural Equation Modeling using AMOS, the results indicated a statistically significant positive relationship between severity, frequency and inhibited sexual disturbances but found no direct relationship between physical abuse, the identity of the perpetrator, the duration of the abuse and inhibited sexual disturbances. Results also indicated that shame had a direct positive relationship to inhibited sexual disturbances. The third finding was that physical abuse and severity of abuse had a significant positive relationship with shame which implies that shame is a moderating variable for inhibited sexual disturbances in adult women survivors of childhood sexual abuse. Internalized shame may be an important factor for therapists to consider in helping survivors overcome sexual inhibition as a result of childhood sexual abuse. If a woman develops a shame-prone identity she may be at risk for experiencing inhibition in her sexuality.
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Perceptions about the O Icheke Multiple Concurrent Parnership Campaign among young people who are members of the Selebi Phikwe District Youth Council, BotswanaMatlapeng, Kgosiekae Maxwell January 2014 (has links)
The purpose of this study was to investigate the perceptions of young people regarding the O Icheke MCP campaign who are members of Selebi Phikwe District Youth Council, in a mining town in central Botswana. The main objective of the study was to explore young people‘s views about MCP, to solicit their views on the key strength and weaknesses of the campaign, and to make recommendations to further strengthen the campaign.
Within the framework of the Health Belief Model, a qualitative research approach was employed and data was collected using four focus group discussions categorised into the following strata
Males aged 18-24 years
Females aged 18-24 years
Males aged 25-35 years
Females aged 25-35 years
The overall study finding was that financial issues, the apparent need for sexual variety, quest for material possession, and the impact of unemployment, migration and alcohol abuse are seen as some of the major factors underlying MCP in Botswana. The O Icheke Multiple Concurrent Partnership campaign was positively viewed as playing a key role in addressing the apparent knowledge gap regarding the impacts of these partnerships and in changing people‘s attitudes towards MCP and discouraging wide sexual networks.
The campaign‘s target population, funding, stakeholders and delivery mode of the program were identified as its pillars and an effective vehicle to achieve its goals. The consultation between funding office and implementing organisations, coverage of the programme in surrounding areas and growing non-profit organisation were identified to affect the MCP campaign to achieve its goals. The recommendation is that more has to be done to improve the implementation of O Icheke programme through more funding, consultation with programme implementers, tailor made for people with disability and covering surrounding areas.
The recommendation is that more has to be done to improve the implementation of O Icheke programme through more funding, consultation and capacity building of volunteers. / Health Studies / MA (Social Behaviour Studies in HIV and AIDS)
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Perceptions about the O Icheke Multiple Concurrent Parnership Campaign among young people who are members of the Selebi Phikwe District Youth Council, BotswanaMatlapeng, Kgosiekae Maxwell 12 June 2014 (has links)
The purpose of this study was to investigate the perceptions of young people regarding the O Icheke MCP campaign who are members of Selebi Phikwe District Youth Council, in a mining town in central Botswana. The main objective of the study was to explore young people‘s views about MCP, to solicit their views on the key strength and weaknesses of the campaign, and to make recommendations to further strengthen the campaign.
Within the framework of the Health Belief Model, a qualitative research approach was employed and data was collected using four focus group discussions categorised into the following strata
Males aged 18-24 years
Females aged 18-24 years
Males aged 25-35 years
Females aged 25-35 years
The overall study finding was that financial issues, the apparent need for sexual variety, quest for material possession, and the impact of unemployment, migration and alcohol abuse are seen as some of the major factors underlying MCP in Botswana. The O Icheke Multiple Concurrent Partnership campaign was positively viewed as playing a key role in addressing the apparent knowledge gap regarding the impacts of these partnerships and in changing people‘s attitudes towards MCP and discouraging wide sexual networks.
The campaign‘s target population, funding, stakeholders and delivery mode of the program were identified as its pillars and an effective vehicle to achieve its goals. The consultation between funding office and implementing organisations, coverage of the programme in surrounding areas and growing non-profit organisation were identified to affect the MCP campaign to achieve its goals. The recommendation is that more has to be done to improve the implementation of O Icheke programme through more funding, consultation with programme implementers, tailor made for people with disability and covering surrounding areas.
The recommendation is that more has to be done to improve the implementation of O Icheke programme through more funding, consultation and capacity building of volunteers. / Health Studies / M.A. (Social Behaviour Studies in HIV and AIDS)
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