Matching Items (148)
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Description
Serial femtosecond X-ray crystallography (SFX) using an X-ray free electron laser (XFEL) is a recent advancement in structural biology for solving crystal structures of challenging membrane proteins, including G-protein coupled receptors (GPCRs), which often only produce microcrystals. An XFEL delivers highly intense X-ray pulses of femtosecond duration short enough to

Serial femtosecond X-ray crystallography (SFX) using an X-ray free electron laser (XFEL) is a recent advancement in structural biology for solving crystal structures of challenging membrane proteins, including G-protein coupled receptors (GPCRs), which often only produce microcrystals. An XFEL delivers highly intense X-ray pulses of femtosecond duration short enough to enable the collection of single diffraction images before significant radiation damage to crystals sets in. Here we report the deposition of the XFEL data and provide further details on crystallization, XFEL data collection and analysis, structure determination, and the validation of the structural model. The rhodopsin-arrestin crystal structure solved with SFX represents the first near-atomic resolution structure of a GPCR-arrestin complex, provides structural insights into understanding of arrestin-mediated GPCR signaling, and demonstrates the great potential of this SFX-XFEL technology for accelerating crystal structure determination of challenging proteins and protein complexes.
ContributorsZhou, X. Edward (Author) / Gao, Xiang (Author) / Barty, Anton (Author) / Kang, Yanyong (Author) / He, Yuanzheng (Author) / Liu, Wei (Author) / Ishchenko, Andrii (Author) / White, Thomas A. (Author) / Yefanov, Oleksandr (Author) / Han, Gye Won (Author) / Xu, Qingping (Author) / de Waal, Parker W. (Author) / Suino-Powell, Kelly M. (Author) / Boutet, Sebastien (Author) / Williams, Garth J. (Author) / Wang, Meitian (Author) / Li, Dianfan (Author) / Caffrey, Martin (Author) / Chapman, Henry N. (Author) / Spence, John (Author) / Fromme, Petra (Author) / Weierstall, Uwe (Author) / Stevens, Raymond C. (Author) / Cherezov, Vadim (Author) / Melcher, Karsten (Author) / Xu, H. Eric (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2016-04-12
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Description
Diacylglycerol kinase catalyses the ATP-dependent conversion of diacylglycerol to phosphatidic acid in the plasma membrane of Escherichia coli. The small size of this integral membrane trimer, which has 121 residues per subunit, means that available protein must be used economically to craft three catalytic and substrate-binding sites centred about the

Diacylglycerol kinase catalyses the ATP-dependent conversion of diacylglycerol to phosphatidic acid in the plasma membrane of Escherichia coli. The small size of this integral membrane trimer, which has 121 residues per subunit, means that available protein must be used economically to craft three catalytic and substrate-binding sites centred about the membrane/cytosol interface. How nature has accomplished this extraordinary feat is revealed here in a crystal structure of the kinase captured as a ternary complex with bound lipid substrate and an ATP analogue. Residues, identified as essential for activity by mutagenesis, decorate the active site and are rationalized by the ternary structure. The γ-phosphate of the ATP analogue is positioned for direct transfer to the primary hydroxyl of the lipid whose acyl chain is in the membrane. A catalytic mechanism for this unique enzyme is proposed. The active site architecture shows clear evidence of having arisen by convergent evolution.
ContributorsLi, Dianfan (Author) / Stansfeld, Phillip J. (Author) / Sansom, Mark S. P. (Author) / Keogh, Aaron (Author) / Vogeley, Lutz (Author) / Howe, Nicole (Author) / Lyons, Joseph A. (Author) / Aragao, David (Author) / Fromme, Petra (Author) / Fromme, Raimund (Author) / Basu, Shibom (Author) / Grotjohann, Ingo (Author) / Kupitz, Christopher (Author) / Rendek, Kimberley (Author) / Weierstall, Uwe (Author) / Zatsepin, Nadia (Author) / Cherezov, Vadim (Author) / Liu, Wei (Author) / Bandaru, Sateesh (Author) / English, Niall J. (Author) / Gati, Cornelius (Author) / Barty, Anton (Author) / Yefanov, Oleksandr (Author) / Chapman, Henry N. (Author) / Diederichs, Kay (Author) / Messerschmidt, Marc (Author) / Boutet, Sebastien (Author) / Williams, Garth J. (Author) / Seibert, M. Marvin (Author) / Caffrey, Martin (Author) / College of Liberal Arts and Sciences (Contributor) / School of Molecular Sciences (Contributor) / Biodesign Institute (Contributor) / Applied Structural Discovery (Contributor) / Department of Physics (Contributor)
Created2015-12-17
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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

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
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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.

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
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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

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.
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
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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

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
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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

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
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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

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.
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
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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

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.
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
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Description
The importance of second language learning in today’s ever-increasing globalized world is becoming ever more paramount. Despite seeming trends which indicate aversion to globalization, the international phenomenon which describes the connection made between peoples and cultures, will only increase its influence in coming years. With the advances globalization has made,

The importance of second language learning in today’s ever-increasing globalized world is becoming ever more paramount. Despite seeming trends which indicate aversion to globalization, the international phenomenon which describes the connection made between peoples and cultures, will only increase its influence in coming years. With the advances globalization has made, it is becoming more important to learn and study foreign languages in order to keep abreast of this trend and not be left behind by globalization. Why electronic translation is not so viable in the long run (at least currently) is that culture and syntax are not things which can be simply ascertained via mediums such as application use. Due to the fact that advanced language proficiency is considered to be an integral piece towards stronger sentiments of “integration” (i.e. Syrian refugees integrating into EU and US) it is of more importance that increasing second language proficiency receives the adequate study and implementation to reflect a more cohesive globalized world. Accompanying this necessity is the simple fact that adult second language learners often struggle to overcome difficulty to the challenging yet rewarding task of learning and eventually mastering a second language.

To truly understand the difficulty some adult second language learners have with learning a second language it can be helpful to compare second language acquisition to how one naturally, and seemingly effortlessly in many cases, acquires their native language. How can a comparative analysis of how native speakers and adult second language learners each learn their first and second language respectively be successfully converted to a specific means of assisting adult second language learners achieve the highest level of possible fluency? In order to more accurately propose a viable solution to the overarching question, the following three questions have been proposed as a means to springboard into better understanding the nature of the main topic.
The points to consider while analyzing the main question throughout this analysis are as follows: How would it be best for an adult second language learner to achieve the same level off proficiency as a native speaker of a given language whom has been exposed to all of its intricacies since birth? At what point exactly is someone considered to have the same level of proficiency which a native speaker of a given language would have and how does that differ from being a heritage speaker? With the final supporting question being: What type of learning would be best suited in helping a heritage speaker (someone who learns a language in the home by virtue of their heritage) or adult second language learner to become highly proficient in a second language?
In order to propose a wide variety of integration between these questions with the conjoined purpose of answering the inquiry of this thesis, many different sources supporting each of the above questions will contain certain overlap, providing a clear basis for constructing a tri-fold argument in answering the thesis question as acutely as possible. In regards to the first question, the question of “proficiency” will be a subsection committed to understanding the nature of how language proficiency works and at what point (if ever) one can ever be considered “highly proficient” in a second language.

All three exploratory questions are compatible with a theory known as Critical Period Theory, a theory in linguistics proposed by Montreal neurologist Wilder Penfield, which states, “There is a critical age, before puberty, that one must learn language. If one has not learned to speak before puberty it is much more difficult, and sometimes impossible, to learn language and speak in a meaningful way.” The overlap which this method binds to Universal Grammar is a rather close-knit relationship. Research composed by certain linguists suggest that “children are born with a certain universal grammar wired into their brains.” This will be compared and cross-examined to a higher degree in a later section of the paper.

The importance of Universal Grammar in relation to Critical Period Theory cannot be overstated.  Universal Grammar in relation to the second language Critical Period Theory will help explain at which point, someone is considered to be a “native speaker” of a given language. The third question posed of how would it be best for heritage and second language learners to increase their proficiency in a second language really touches on both of these theories in regards to at which age someone is exposed to a specific language in addition to how Universal Grammar affects the development of second language acquisition. In the realm of perpetually working towards mitigating an answer to this analysis’s thesis, the connecting thread or “roter Faden”, as it is said in German, will be the integrative domain the above questions will have on arriving to a clearer understanding of the nature of how a comparative analysis of second language learners and mother-tongue speakers can expedite the language learning process of second language learners, using techniques of native speakers which they inherently pick-up.
Created2018-12