Matching Items (127)
Description
Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
ContributorsEdlund, Petra (Author) / Takala, Heikki (Author) / Claesson, Elin (Author) / Henry, Leocadie (Author) / Dods, Robert (Author) / Lehtivuori, Heli (Author) / Panman, Matthijs (Author) / Pande, Kanupriya (Author) / White, Thomas (Author) / Nakane, Takanori (Author) / Berntsson, Oskar (Author) / Gustavsson, Emil (Author) / Bath, Petra (Author) / Modi, Vaibhav (Author) / Roy Chowdhury, Shatabdi (Author) / Zook, James (Author) / Berntsen, Peter (Author) / Pandey, Suraj (Author) / Poudyal, Ishwor (Author) / Tenboer, Jason (Author) / Kupitz, Christopher (Author) / Barty, Anton (Author) / Fromme, Petra (Author) / Koralek, Jake D. (Author) / Tanaka, Tomoyuki (Author) / Spence, John (Author) / Liang, Mengning (Author) / Hunter, Mark S. (Author) / Boutet, Sebastien (Author) / Nango, Eriko (Author) / Moffat, Keith (Author) / Groenhof, Gerrit (Author) / Ihalainen, Janne (Author) / Stojkovic, Emina A. (Author) / Schmidt, Marius (Author) / Westenhoff, Sebastian (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2016-10-19
Description
Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1 mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.
ContributorsNogly, Przemyslaw (Author) / Panneels, Valerie (Author) / Nelson, Garrett (Author) / Gati, Cornelius (Author) / Kimura, Tetsunari (Author) / Milne, Christopher (Author) / Milathianaki, Despina (Author) / Kubo, Minoru (Author) / Wu, Wenting (Author) / Conrad, Chelsie (Author) / Coe, Jesse (Author) / Bean, Richard (Author) / Zhao, Yun (Author) / Bath, Petra (Author) / Dods, Robert (Author) / Harimoorthy, Rajiv (Author) / Beyerlein, Kenneth R. (Author) / Rheinberger, Jan (Author) / James, Daniel (Author) / Deponte, Daniel (Author) / Li, Chufeng (Author) / Sala, Leonardo (Author) / Williams, Garth J. (Author) / Hunter, Mark S. (Author) / Koglin, Jason E. (Author) / Berntsen, Peter (Author) / Nango, Eriko (Author) / Iwata, So (Author) / Chapman, Henry N. (Author) / Fromme, Petra (Author) / Frank, Matthias (Author) / Abela, Rafael (Author) / Boutet, Sebastien (Author) / Barty, Anton (Author) / White, Thomas A. (Author) / Weierstall, Uwe (Author) / Spence, John (Author) / Neutze, Richard (Author) / Schertler, Gebhard (Author) / Standfuss, Jorg (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor) / Department of Chemistry and Biochemistry (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / School of Molecular Sciences (Contributor)
Created2016-08-22
Description
Dissolved organic matter (DOM) can have numerous effects on the water chemistry and the biological life within an aquatic system with its wide variety of chemical structures and properties. The composition of the dissolved carbon can be estimated by utilizing the fluorescent properties of some DOM such as aromatic amino acids and humic material. This experiment was used to observe how organic matter could influence hydrothermal systems, such as Sylvan Springs in Yellowstone National Park, USA. Using optical density at 600 nm (OD 600), excitation-emission matrix spectra (EEMS), and Illumina sequencing methods (16S rRNA gene sequencing), changes in dissolved organic matter (DOM) were observed based on long term incubation at 84ºC and microbial influence. Four media conditions were tested over a two-month duration to assess these changes: inoculated pine needle media, uninoculated pine needle media, inoculated yeast extract media, and uninoculated yeast extract media. The inoculated samples contained microbes from a fluid and sediment sample of Sylvan Spring collected July 23, 2018. Absorbance indicated that media containing pine needle broth poorly support life, whereas media containing yeast extract revealed a positive increase in growth. Excitation-Emission Matrix Spectra of the all media conditions indicated changes in DOM composition throughout the trial. There were limited differences between the inoculated and uninoculated samples suggesting that the DOM composition change in this study was dominated by the two-month incubation at 84ºC more than biotic processes. Sequencing performed on a sediment sample collected from Sylvan Spring indicated five main order of prokaryotic phyla: Aquificales, Desulfurococcales, Thermoproteales, Thermodesulfobacteriales, and Crenarchaeota. These organisms are not regarded as heterotrophic microbes, so the lack of significant biotic changes in DOM could be a result of these microorganisms not being able to utilize these enrichments as their main metabolic energy supply.
ContributorsKnott, Nicholas Joseph (Author) / Shock, Everett (Thesis director) / Hartnett, Hilairy (Committee member) / Till, Christy (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Introduction: There is currently a lack of industry-wide gold standardization in accelerometer study
protocols, including within sleep-focused studies. This study seeks to address accuracy of
accelerometer data in detection of the beginnings and ends of sleep bouts in young adults with
polysomnography (PSG) corroboration. An existing algorithm used to differentiate valid/invalid wear
time and detect bouts of sleep has been modified with the goal of maximizing accuracy of sleep bout
detection. Methods: Three key decisions and thresholds of the algorithm have been modified with three
experimental values each being tested. The main experimental variable Sleepwindow controls the
amount of time before and after a determined bout of sleep that is searched for additional sedentary
time to incorporate and consider part of the same sleep bout. Results were compared to PSG and sleep
diary data for absolute agreement of sleep bout start time (START), end time (END) and time in bed
(TIB). Adjustments were made for outliers as well as sleep latency, snooze time, and the sum of both.
Results: Only adjustments made to a sleep window variable yielded altered results. Between a 5-, 15-,
and 30-minute window, a 15-minute window incurred the least error and most agreement to
comparisons for START, while a 5-minute window was best for END and TIB. Discussion: Contrary
to expectation, corrections for snooze, latency, and both did not substantially improve agreement to
PSG. Algorithm-derived estimates of START and END always fell after sleep diary and PSG both,
suggesting either participants’ sedentary behavior beginning and ends were at a delay from sleep and
wake times, or the algorithm estimates consistently later times than appropriate. The inclusion of a
sleep window variable yields substantial variety in results. A 15-minute window appears best at
determining START while a 5-minute window appears best for END and TIB. Further investigation on
the optimal window length per demographic and condition is required.
protocols, including within sleep-focused studies. This study seeks to address accuracy of
accelerometer data in detection of the beginnings and ends of sleep bouts in young adults with
polysomnography (PSG) corroboration. An existing algorithm used to differentiate valid/invalid wear
time and detect bouts of sleep has been modified with the goal of maximizing accuracy of sleep bout
detection. Methods: Three key decisions and thresholds of the algorithm have been modified with three
experimental values each being tested. The main experimental variable Sleepwindow controls the
amount of time before and after a determined bout of sleep that is searched for additional sedentary
time to incorporate and consider part of the same sleep bout. Results were compared to PSG and sleep
diary data for absolute agreement of sleep bout start time (START), end time (END) and time in bed
(TIB). Adjustments were made for outliers as well as sleep latency, snooze time, and the sum of both.
Results: Only adjustments made to a sleep window variable yielded altered results. Between a 5-, 15-,
and 30-minute window, a 15-minute window incurred the least error and most agreement to
comparisons for START, while a 5-minute window was best for END and TIB. Discussion: Contrary
to expectation, corrections for snooze, latency, and both did not substantially improve agreement to
PSG. Algorithm-derived estimates of START and END always fell after sleep diary and PSG both,
suggesting either participants’ sedentary behavior beginning and ends were at a delay from sleep and
wake times, or the algorithm estimates consistently later times than appropriate. The inclusion of a
sleep window variable yields substantial variety in results. A 15-minute window appears best at
determining START while a 5-minute window appears best for END and TIB. Further investigation on
the optimal window length per demographic and condition is required.
ContributorsMartin, Logan Rhett (Author) / Buman, Matthew (Thesis director) / Toledo, Meynard John (Committee member) / Kurka, Jonathan (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2019-12
Description
This thesis paper examines the effects of increased standing and light physical activity in the workplace on postprandial glucose. Sedentary behavior is detrimental to our health, affecting metabolic risk factors. An easy way to implement change is by decreasing sedentary time in workplaces where sitting is common, such as office workspaces. To consider how postprandial glucose is affected by decreasing sedentary time, participants ate a standardized meal for lunch and were asked to decrease their sitting time by replacing it with standing and light physical activity.
ContributorsChilders, Autumn Skye (Author) / Buman, Matthew (Thesis director) / Sears, Dorothy (Committee member) / Hasanaj, Kristina (Committee member) / College of Health Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2020-12
Description
The purpose of this study was to examine the overall maintenance of behavior during the 12 to 24 month period of the Stand&Move@Work study and the impact of implementation factors (i.e., facilitators, advocate activity, and the amount of strategies used) on behavior change. The design of the study was a cluster randomized trial which was facilitated by researchers for the first 12 months of the study. The primary aim of the study was to examine the maintenance of behavior change (i.e., sitting time) at the 12 month and 24 month marks using objectively measured sedentary behavior (activPAL micro). The secondary aim of the study was to examine the impact of implementation factors (i.e., facilitators, advocate activity, and the amount of strategies used) on behavior change during the 12 through 24 months maintenance period. Participants (N=630) included full-time, caucasian, middle-aged office workers. For the primary aim, descriptive means were used to cluster for observations within-persons and were adjusted for age, gender, race, job-type, and ordering effects.. For the secondary aim, descriptive means adjusted for workplace culture and environment were computed. At the 24 month mark, participants spent 280.67 ± 87.67 min/8hr workday sitting and 161.94 ± 85.87 min/8hr workday standing. The top performing worksites displayed reductions in sitting time which largely translated into standing time by about 30 minutes per 8 hour workday at 24 months. Feasibility findings indicated that implementation strategies do not show differences between the top 25% and bottom 25% performing worksites. This study provides insight to implementation strategies for interventions in the workplace.
ContributorsTong, Alyssa Taylor (Author) / Buman, Matthew (Thesis director) / Larouche, Miranda (Committee member) / Estabrooks, Paul (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Cardiovascular disease attributed to about 800,000 deaths per year and is the leading cause of all-cause mortality in the U.S. Previous studies indicate that reducing sedentary time or increasing physical activity (PA) can independently reduce cardiometabolic risk (CMR). Further, studies have shown that higher levels of moderate-to-vigorous PA can attenuate the negative effects of sedentary behavior on CMR.
In this study, we evaluated the association between sedentary time, light-intensity PA (LPA), and moderate-vigorous PA (MVPA) and CMR biomarkers (high density lipoprotein level, low density lipoprotein level, triglycerides, fasting glucose, total cholesterol, blood pressure, and body mass index). Additionally, we examined if the detrimental association between sedentary time and CMR biomarkers is partially or fully attenuated by MVPA. Baseline objective physical activity and cardiometabolic risk data from a two-arm-cluster randomized trial (Stand&Move@work) were used in this study. Multilevel models clustered by worksite evaluated the fixed effects and interaction between MVPA and sedentary time on CMR. Data from 630 sedentary working adults (from 24 worksites) were included in the analysis. The sample was mainly middle aged (44.6±11.2) females (74%) with race distributions as follows; 70.5% white, 13.8% hispanic, 4.1% black, 5.1% asian, and 2.1% other. Our study showed detrimental trends consistent with previous studies between sedentary behavior and cardiometabolic outcomes including HDL, LDL, and total cholesterol. MVPA demonstrated beneficial associations with lipoproteins including HDL, LDL, total cholesterol, and triglycerides. We observed that high levels of MVPA may be able to partially attenuate the negative effects of highly sedentary behavior on fasting glucose, total cholesterol, and LDL levels. Overall, sedentary behavior indicated deleterious associations with cardiometabolic outcomes. Future directions for this study could examine a more at-risk population or a highly active population for further assessment of CMR biomarkers and the effects of behavior.
In this study, we evaluated the association between sedentary time, light-intensity PA (LPA), and moderate-vigorous PA (MVPA) and CMR biomarkers (high density lipoprotein level, low density lipoprotein level, triglycerides, fasting glucose, total cholesterol, blood pressure, and body mass index). Additionally, we examined if the detrimental association between sedentary time and CMR biomarkers is partially or fully attenuated by MVPA. Baseline objective physical activity and cardiometabolic risk data from a two-arm-cluster randomized trial (Stand&Move@work) were used in this study. Multilevel models clustered by worksite evaluated the fixed effects and interaction between MVPA and sedentary time on CMR. Data from 630 sedentary working adults (from 24 worksites) were included in the analysis. The sample was mainly middle aged (44.6±11.2) females (74%) with race distributions as follows; 70.5% white, 13.8% hispanic, 4.1% black, 5.1% asian, and 2.1% other. Our study showed detrimental trends consistent with previous studies between sedentary behavior and cardiometabolic outcomes including HDL, LDL, and total cholesterol. MVPA demonstrated beneficial associations with lipoproteins including HDL, LDL, total cholesterol, and triglycerides. We observed that high levels of MVPA may be able to partially attenuate the negative effects of highly sedentary behavior on fasting glucose, total cholesterol, and LDL levels. Overall, sedentary behavior indicated deleterious associations with cardiometabolic outcomes. Future directions for this study could examine a more at-risk population or a highly active population for further assessment of CMR biomarkers and the effects of behavior.
ContributorsMeyer, Emily Camille (Author) / Buman, Matthew (Thesis director) / Toledo, Meynard (Committee member) / Pereira, Mark (Committee member) / School of Life Sciences (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
While type 2 diabetes (T2D) rates have soared, the number of Americans classified as ‘prediabetic’ has also increased. Despite this, current preventative approaches are costly and often not without undue side-effects. Instead, behavioral lifestyle approaches hold promise in reducing conversion rates of T2D as the latest treatment option that could mitigate and transform disease management. However, present interventions do not possess the scope necessary for implementation in a realistic, scalable way that can target the large at-risk population.
The application (app) “BeWell24” mitigates this diabetes risk through targeting sleep, physical activity, sedentary behavior, and diet, and is being delivered through mHealth technology to attenuate the higher-risk of the prediabetic Veteran population. In order for full scale dissemination, this thesis examines a provider perspective of the ‘Post-intervention interview guide’, performed with a Phoenix Veterans Affairs Health Care System (PVAHCS) provider. It then suggests revisions to the interview guide based on the provider’s interview and existing literature. This thesis also emphasizes the rationale behind these proposed changes to be organized in line with the iPARIHS framework (integrated Promoting Action on Research Implementation in Health Services).
Overall, the provider responded positively to BeWell24 and the ‘Post-intervention interview guide’, with constructive suggestions for each question in the interview guide. The main theme of the provider’s answers and comments were to prioritize efficiency and preserve standard clinical flow. A revised interview guide is provided, which prospectively presents as a more brief and focused interview organized by the iPARIHS framework. This revised interview guide could aid in the clarity of provider responses, specifically for the prospective interviews of the ongoing larger BeWell24 study and future studies.
The application (app) “BeWell24” mitigates this diabetes risk through targeting sleep, physical activity, sedentary behavior, and diet, and is being delivered through mHealth technology to attenuate the higher-risk of the prediabetic Veteran population. In order for full scale dissemination, this thesis examines a provider perspective of the ‘Post-intervention interview guide’, performed with a Phoenix Veterans Affairs Health Care System (PVAHCS) provider. It then suggests revisions to the interview guide based on the provider’s interview and existing literature. This thesis also emphasizes the rationale behind these proposed changes to be organized in line with the iPARIHS framework (integrated Promoting Action on Research Implementation in Health Services).
Overall, the provider responded positively to BeWell24 and the ‘Post-intervention interview guide’, with constructive suggestions for each question in the interview guide. The main theme of the provider’s answers and comments were to prioritize efficiency and preserve standard clinical flow. A revised interview guide is provided, which prospectively presents as a more brief and focused interview organized by the iPARIHS framework. This revised interview guide could aid in the clarity of provider responses, specifically for the prospective interviews of the ongoing larger BeWell24 study and future studies.
ContributorsWojtas, Abby Ann (Author) / Buman, Matthew (Thesis director) / Larouche, Miranda (Committee member) / Epstein, Dana (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Mobile health or "mHealth" defines a broad spectrum of medical or public health practice supported by mobile devices. The patient's perception of mobile health applications is the key point in confronting whether or not patients will utilize the tools at their disposal As such, the primary aim of this study was to examine participant feedback through quantitative and qualitative measures using the Therapy Evaluation Questionnaire and a patient interview, respectively, to further understand the patient rated acceptability of using BeWell24 and SleepWell24 for improving health outcomes. For BeWell24, it was hypothesized that patients who received the Multicomponent version would report higher acceptability scores than those randomized to the Health Education version. Furthermore, in regard to SleepWell24, it was hypothesized that the SleepWell24 patient would provide positive feedback and suggestions regarding their own experience with the SleepWell24 app. Data from this thesis was pulled from two ongoing randomized controlled trials currently being conducted at the Phoenix Veteran Affairs Health Care Service (PVACHS) and Mayo Clinic hospitals. Means, standard deviations, frequencies, and percentages were commuted to summarize demographics and TEQ scores. In addition, key concepts from a qualitative interview with a SleepWell24 participant were derived. The results showed a greater acceptability of the multicomponent versions of BeWell24 and SleepWell24 but a lower TEQ score of perceived usability. mHealth implementations pose a potential to become an important part of the health sector for establishing innovative approaches to delivering care, and while benefits have been highly praised, it is clear that the perceptions of mHealth must be positive if the technology is to transcend into a practical clinical setting.
ContributorsJimenez, Asael (Author) / Buman, Matthew (Thesis director) / Epstein, Dana (Committee member) / School of Nutrition and Health Promotion (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
Finding life beyond Earth could change our understanding of life and habitability. The best place to look for life beyond Earth is Jupiter's moon, Europa. It has been estimated Europa may have a liquid, salt-water subsurface with 2 to 3 times the volume of all Earth's oceans. Knowing that all life requires water, it is in our best interest to explore Europa. This thesis explored the plausibility of life on Europa in four of its environments: on the surface, under the ice shell, in the liquid subsurface, and at the bottom of the liquid subsurface. Each of these environments were defined from science literature and compared to known Earth analogs. Europa's surface is not likely to support life, as there is not liquid water present. There is also extremely high radiation bombardment and extremely low surface temperatures that are estimated to be well out of the range for supporting life. It is more plausible that life could be under Europa's ice shell than on the surface. Under the surface, radiation exposure dramatically reduces. Researchers have found organisms on Earth that can live in similar environments as Europa's ice as well. These organisms require some interaction with liquid water though. Uncertainties about Europa's ice shell thickness and radiation load per depth it experiences, as well as there being limited research on organisms in ice environments, hinder us from definitively assessing the plausibility of life under the surface. The best environment on Europa to look for life on Europa is the subsurface. There remain a lot of uncertainties about the subsurface, however, that make it difficult to assess the plausibility of finding life. These uncertainties include its depth, water activity, salinity, temperature, pressure, and structure. This subsurface may be suitable for life, but until we can further understand the environment of the subsurface, we cannot make definite conclusions. As for assessing the plausibility of life at the bottom of Europa's subsurface, there is not much we know about this environment either. It has been suggested there may be hydrothermal vents, but no evidence has either supported or rejected this idea. Without a clear understanding of the environment at the bottom of the subsurface, the plausibility of life here cannot be definitively answered. It is apparent we need to further study Europa. In particular, we need to focus on understanding the subsurface. When the subsurface is better defined, we can better assess the plausibility of life being present. Fortunately, both NASA and the ESA are currently planning missions to Europa that are scheduled to launch in the 2020s.
ContributorsHoward, Cheyenne Whiffen (Author) / Farmer, Jack (Thesis director) / Shock, Everett (Committee member) / School of Earth and Space Exploration (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05