A new solution to the curved Ewald sphere problem for 3D image reconstruction in electron microscopy.

We develop an algorithm capable of imaging a three-dimensional object given a collection of two-dimensional images of that object that are significantly influenced by the curvature of the Ewald sphere. These two-dimensional images cannot be approximated as projections of the object. Such an algorithm is useful in cryo-electron microscopy where larger samples, higher resolution, or lower energy electron beams are desired, all of which contribute to the significance of Ewald curvature.

Quantitative agreement between dynamical rocking curves in ultrafast electron diffraction for x-ray lasers.

Electron diffraction through a thin patterned silicon membrane can be used to create complex spatial modulations in electron distributions. By precisely varying parameters such as crystallographic orientation and wafer thickness, the intensity of reflections in the diffraction plane can be controlled and by placing an aperture to block all but one spot, we can form an image with different parts of the patterned membrane, as is done for bright-field imaging in microscopy. The patterned electron beams can then be used to control phase and amplitude of subsequent x-ray emission, enabling novel coherent x-ray methods. The electrons themselves can also be used for femtosecond time resolved diffraction and microscopy. As a first step toward patterned beams, we demonstrate experimentally and through simulation the ability to accurately predict and control diffraction spot intensities. We simulate MeV transmission electron diffraction patterns using the multislice method for various crystallographic orientations of a single crystal Si(001) membrane near beam normal. The resulting intensity maps of the Bragg reflections are compared to experimental results obtained at the Accelerator Structure Test Area Ultrafast Electron Diffraction (ASTA UED) facility at SLAC. Furthermore, the fraction of inelastic and elastic scattering of the initial charge is estimated along with the absorption of the membrane to determine the contrast that would be seen in a patterned version of the Si(001) membrane.

Dynamics of rare gas solids irradiated by electron beams.

The remarkable success of x-ray free-electron lasers and their ability to image biological macromolecules while outrunning secondary radiation damage due to photoelectrons, by using femtosecond pulses, raise the question of whether this can be done using pulsed high-energy electron beams. In this paper, we use excited state molecular dynamics simulations, with tabulated potentials, for rare gas solids to investigate the effect of radiation damage due to inelastic scattering (by plasmons, excitons, and heat) on the pair distribution function. We use electron energy loss spectra to characterize the electronic excitations responsible for radiation damage.

The short-term effects of a mass reach physical activity campaign: an evaluation using hierarchy of effects model and intention profiles.

BACKGROUND: Mass reach physical activity campaigns are designed to deliver physical-activity related messages to a large population across different media including print, television, radio, and websites. Few evaluations have examined the short-term effects of a mass reach campaign on participants who were engaged with the campaign. The current research examined the short-term effects of the ParticipACTION 150 Play List, a mass reach physical activity campaign, on participants who registered with the campaign website. METHODS: Participants (N = 7801) completed a registration questionnaire measuring demographic information, awareness and recall of physical activity and sport advertising, and self-reported number of activities tried or planned to try from the 150 Play List. A follow-up survey was completed by 1298 participants from the original sample. Additional questions assessed experience with the 150 Play List and attitudes towards campaign advertisements. RESULTS: Approximately 14.5% of participants cited the ParticipACTION 150 Play List and 23.6% mentioned a 'getting active' message when recalling advertisements. Those who named the 150 Play List or getting active reported more activities tried and more activities planned than those who did not. They were also more likely to say they had tried a new activity and planned ongoing participation. It was also found that participants with a disability were more likely to have tried a new activity compared to those not in a minority group. Other correlates of trying new activities at follow-up were younger age, more positive reported experience with the 150 Play List, and more favourable attitudes towards campaign advertisements. Those who did not intend continued participation, or who were unsure at baseline and then decided against continued participation at follow-up, reported they were less sedentary or encouraging others to be active. CONCLUSIONS: This research addresses the gap in evidence regarding the efficacy of mass reach physical activity campaigns by informing whether a year-long campaign like the 150 Play List can be effective in influencing the behavior of those engaged with the campaign. The results reinforce the idea that 'top of mind' awareness should be measured. Investigating intention profiles can help inform campaign impacts and continuation intentions.

Short-wavelength free-electron laser sources and science: a review.

This review is focused on free-electron lasers (FELs) in the hard to soft x-ray regime. The aim is to provide newcomers to the area with insights into: the basic physics of FELs, the qualities of the radiation they produce, the challenges of transmitting that radiation to end users and the diversity of current scientific applications. Initial consideration is given to FEL theory in order to provide the foundation for discussion of FEL output properties and the technical challenges of short-wavelength FELs. This is followed by an overview of existing x-ray FEL facilities, future facilities and FEL frontiers. To provide a context for information in the above sections, a detailed comparison of the photon pulse characteristics of FEL sources with those of other sources of high brightness x-rays is made. A brief summary of FEL beamline design and photon diagnostics then precedes an overview of FEL scientific applications. Recent highlights are covered in sections on structural biology, atomic and molecular physics, photochemistry, non-linear spectroscopy, shock physics, solid density plasmas. A short industrial perspective is also included to emphasise potential in this area.

XFELs for structure and dynamics in biology.

The development and application of the free-electron X-ray laser (XFEL) to structure and dynamics in biology since its inception in 2009 are reviewed. The research opportunities which result from the ability to outrun most radiation-damage effects are outlined, and some grand challenges are suggested. By avoiding the need to cool samples to minimize damage, the XFEL has permitted atomic resolution imaging of molecular processes on the 100 fs timescale under near-physiological conditions and in the correct thermal bath in which molecular machines operate. Radiation damage, comparisons of XFEL and synchrotron work, single-particle diffraction, fast solution scattering, pump-probe studies on photosensitive proteins, mix-and-inject experiments, caged molecules, pH jump and other reaction-initiation methods, and the study of molecular machines are all discussed. Sample-delivery methods and data-analysis algorithms for the various modes, from serial femtosecond crystallo-graphy to fast solution scattering, fluctuation X-ray scattering, mixing jet experiments and single-particle diffraction, are also reviewed.

Structures of riboswitch RNA reaction states by mix-and-inject XFEL serial crystallography.

Riboswitches are structural RNA elements that are generally located in the 5' untranslated region of messenger RNA. During regulation of gene expression, ligand binding to the aptamer domain of a riboswitch triggers a signal to the downstream expression platform. A complete understanding of the structural basis of this mechanism requires the ability to study structural changes over time. Here we use femtosecond X-ray free electron laser (XFEL) pulses to obtain structural measurements from crystals so small that diffusion of a ligand can be timed to initiate a reaction before diffraction. We demonstrate this approach by determining four structures of the adenine riboswitch aptamer domain during the course of a reaction, involving two unbound apo structures, one ligand-bound intermediate, and the final ligand-bound conformation. These structures support a reaction mechanism model with at least four states and illustrate the structural basis of signal transmission. The three-way junction and the P1 switch helix of the two apo conformers are notably different from those in the ligand-bound conformation. Our time-resolved crystallographic measurements with a 10-second delay captured the structure of an intermediate with changes in the binding pocket that accommodate the ligand. With at least a 10-minute delay, the RNA molecules were fully converted to the ligand-bound state, in which the substantial conformational changes resulted in conversion of the space group. Such notable changes in crystallo highlight the important opportunities that micro- and nanocrystals may offer in these and similar time-resolved diffraction studies. Together, these results demonstrate the potential of 'mix-and-inject' time-resolved serial crystallography to study biochemically important interactions between biomacromolecules and ligands, including those that involve large conformational changes.

How perceptions of community environment influence health behaviours: using the Analysis Grid for Environments Linked to Obesity Framework as a mechanism for exploration. / Perceptions à l'égard de l'environnement et comportements en matière de santé : étude fondée sur le cadre ANGELO (grille d'analyse des environnements liés à l'obésité).

INTRODUCTION: Overweight and obesity are influenced by a complex interplay of individual and environmental factors that affect physical activity and healthy eating. Nevertheless, little has been reported on people's perceptions of those factors. Addressing this critical gap and community partner needs, this study explored how people perceived the influence of micro- and macroenvironmental factors on physical activity and healthy eating. METHODS: Community partners wanted the study results in a format that would be readily and easily used by local decision makers. We used photovoice to engage 35 community members across four municipalities in Alberta, Canada, and to share their narratives about their physical activity and healthy eating. A combination of inductive and deductive analysis categorized data by environmental level (micro vs. macro) and type (physical, political, economic, and sociocultural), guided by the Analysis Grid for Environments Linked to Obesity Framework. RESULTS: Participants conceptualized health-influencing factors more broadly than physical activity and healthy eating to include "community social health." Participants spoke most often about the influence of the microenvironment (n = 792 ANGELO Framework coding tallies) on their physical activity, healthy eating and community social health in comparison to the macroenvironment (n = 93). Photovoice results provided a visual narrative to community partners and decision makers about how people's ability to make healthy choices can be limited by macroenvironmental forces beyond their control. CONCLUSION: Focussing future research on macro- and microenvironmental influences and localized community social health can inform practice by providing strategies on how to implement healthy changes within communities, while ensuring that research and interventions echo diverse people's perceptions.

INTRODUCTION: Le surpoids et l'obésité dépendent de l'interaction complexe entre facteurs liés à l'indivu et facteurs liés à l'environnement ayant une influence sur l'activité physique et une alimentation saine. Il existe pourtant peu d'information sur les perceptions des individus à l'égard de ces facteurs. Afin de combler cette lacune importante et de répondre aux besoins de nos partenaires des collectivités, nous examinons comment les individus perçoivent l'influence des facteurs micro- et macroenvironnementaux sur leur activité physique et leur alimentation. MÉTHODOLOGIE: Nos partenaires des collectivités souhaitaient que les résultats de l'étude soient diffusés dans un format facile à utiliser par les décideurs locaux. Avec la méthode photovoix, nous avons motivé 35 membres issus de quatre collectivités de l'Alberta (Canada) à fournir leurs témoignages à propos de leur activité physique et d'une alimentation saine. Nous avons employé une combinaison d'analyses inductives et déductives pour classer les données par niveau (micro et macro) d'environnement et par type (environnement physique, politique, économique et socioculturel), à l'aide de la Grille d'analyse des environnements liés à l'obésité (ANGELO). RÉSULTATS: Pour les participants, les facteurs liés à la santé étaient plus larges que l'activité physique et une alimentation saine, incluant la « santé sociocommunautaire ¼. Les participants ont parlé plus souvent de l'influence du microenvironnement (score de 792 après codage au moyen du cadre ANGELO) sur l'activité physique, une alimentation saine ou la santé sociocommunautaire que sur l'influence du macroenvironnement (score de 93). Les résultats obtenus avec la méthode photovoix ont fourni aux partenaires des collectivités et aux décideurs un témoignage visuel de la manière dont la capacité des individus à faire des choix sains peut être restreinte par des forces macroenvironnementales qui échappent à leur contrôle. CONCLUSION: Axer les recherches ultérieures sur les influences et macroenvironnementales et microenvironnementales ainsi que sur la santé sociocommunautaire locale sera utile, car cela alimentera les stratégies de changement en vue d'améliorer la santé des collectivités tout en garantissant que la recherche et les interventions font écho aux perceptions diverses de la population.

Exploring women's responses to online media coverage of weight loss surgery.

The purpose of this study was to understand the reactions of women to online news articles about weight loss surgery and related reader comments. Focus groups were conducted; open-ended questions were asked to elicit responses to existing online news media content related to weight loss surgery. The participants described the online articles as predominantly supportive of weight loss surgery and in response they expressed a desire to see more critical content, including different and competing perspectives. Participants felt the online comments represented extreme perspectives and were predominately negative. These were therefore not viewed as helpful or informative. Nevertheless, readers viewed comments as a form of entertainment. Because of the aggressive and anonymous nature of reader comments in response to online news stories, the participants did not feel comfortable leaving comments themselves on the news sites. Findings highlight the importance of gathering readers' perspectives in response to interactive media content and, in particular, health information.

The linac coherent light source single particle imaging road map.

Intense femtosecond x-ray pulses from free-electron laser sources allow the imaging of individual particles in a single shot. Early experiments at the Linac Coherent Light Source (LCLS) have led to rapid progress in the field and, so far, coherent diffractive images have been recorded from biological specimens, aerosols, and quantum systems with a few-tens-of-nanometers resolution. In March 2014, LCLS held a workshop to discuss the scientific and technical challenges for reaching the ultimate goal of atomic resolution with single-shot coherent diffractive imaging. This paper summarizes the workshop findings and presents the roadmap toward reaching atomic resolution, 3D imaging at free-electron laser sources.

Examining the steps-per-day trajectories of cardiac rehabilitation patients: a latent class growth analysis perspective.

PURPOSE: Research suggests that cardiac rehabilitation (CR) patients need to engage in at least 6500 steps per day to obtain health benefits. Unfortunately, very little is known about the steps-per-day trajectories of these patients and whether the demographic, clinical, and CR program characteristics are similar for these trajectories. METHODS: Patients (n = 235) completed a questionnaire assessing demographic and clinical variables upon entry to CR and subsequently wore a pedometer for 7 days at the end of CR, and 3, 6, and 9 months after completing CR. RESULTS: Latent class growth analyses showed that 3 classes of patients emerged that were termed nonadherers (averaged ∼3112 steps per day at the end of CR and remained stable up to 9 months after CR), significant decliners (averaged ∼7010 steps per day at the end of CR and steadily declined after CR), and optimal adherers (averaged ∼10 700 steps per day and remained stable after CR). Logistic regressions showed that nonadherers were more likely to be obese, have at least 1 comorbidity, and a lower exercise capacity compared with the significant decliners/optimal adherers. CONCLUSIONS: Distinct steps-per-day trajectories exist for CR patients that are partially distinguished by demographic and clinical variables.

The extraction of single-particle diffraction patterns from a multiple-particle diffraction pattern.

The structures of biological molecules may soon be determined with X-ray free-electron lasers without crystallization by recording the coherent diffraction patterns of many identical copies of a molecule. Most analysis methods require a measurement of each molecule individually. However, current injection methods deliver particles to the X-ray beam stochastically and the maximum yield of single particle measurements is 37% at optimal concentration. The remaining 63% of pulses intercept no particles or multiple particles. We demonstrate that in the latter case single particle diffraction patterns can be extracted provided the particles are sufficiently separated. The technique has the potential to greatly increase the amount of data available for three-dimensional imaging of identical particles with X-ray lasers.

Lawrence Bragg, microdiffraction and X-ray lasers.

We trace the historical development of W. L. Bragg's `law' and the key experimental observation which made it possible using polychromatic radiation at a time when neither X-ray wavelengths nor cell constants were known. This led, through his phasing and solving large mineral structures (without use of a computer), to work on metals, proteins, bubble rafts and his X-ray microscope. The relationship of this to early X-ray microdiffraction is outlined, followed by a brief review of electron microdiffraction methods, where electron-probe sizes smaller than one unit cell can be formed with an interesting `failure' of Bragg's law. We end with a review of recent femtosecond X-ray `snapshot' diffraction from protein nanocrystals, using an X-ray laser which generates pulses so short that they terminate before radiation damage can commence, yet subsequently destroy the sample. In this way, using short pulses instead of freezing, the nexus between dose, resolution and crystal size has been broken, opening the way to time-resolved diffraction without damage for a stream of identical particles.

Single-particle structure determination by correlations of snapshot X-ray diffraction patterns.

Diffractive imaging with free-electron lasers allows structure determination from ensembles of weakly scattering identical nanoparticles. The ultra-short, ultra-bright X-ray pulses provide snapshots of the randomly oriented particles frozen in time, and terminate before the onset of structural damage. As signal strength diminishes for small particles, the synthesis of a three-dimensional diffraction volume requires simultaneous involvement of all data. Here we report the first application of a three-dimensional spatial frequency correlation analysis to carry out this synthesis from noisy single-particle femtosecond X-ray diffraction patterns of nearly identical samples in random and unknown orientations, collected at the Linac Coherent Light Source. Our demonstration uses unsupported test particles created via aerosol self-assembly, and composed of two polystyrene spheres of equal diameter. The correlation analysis avoids the need for orientation determination entirely. This method may be applied to the structural determination of biological macromolecules in solution.

X-ray lasers for structural and dynamic biology.

Research opportunities and techniques are reviewed for the application of hard x-ray pulsed free-electron lasers (XFEL) to structural biology. These include the imaging of protein nanocrystals, single particles such as viruses, pump--probe experiments for time-resolved nanocrystallography, and snapshot wide-angle x-ray scattering (WAXS) from molecules in solution. The use of femtosecond exposure times, rather than freezing of samples, as a means of minimizing radiation damage is shown to open up new opportunities for the molecular imaging of biochemical reactions at room temperature in solution. This is possible using a 'diffract-and-destroy' mode in which the incident pulse terminates before radiation damage begins. Methods for delivering hundreds of hydrated bioparticles per second (in random orientations) to a pulsed x-ray beam are described. New data analysis approaches are outlined for the correlated fluctuations in fast WAXS, for protein nanocrystals just a few molecules on a side, and for the continuous x-ray scattering from a single virus. Methods for determining the orientation of a molecule from its diffraction pattern are reviewed. Methods for the preparation of protein nanocrystals are also reviewed. New opportunities for solving the phase problem for XFEL data are outlined. A summary of the latest results is given, which now extend to atomic resolution for nanocrystals. Possibilities for time-resolved chemistry using fast WAXS (solution scattering) from mixtures is reviewed, toward the general goal of making molecular movies of biochemical processes.

Injector for scattering measurements on fully solvated biospecies.

We describe a liquid jet injector system developed to deliver fully solvated microscopic target species into a probe beam under either vacuum or ambient conditions. The injector was designed specifically for x-ray scattering studies of biological nanospecies using x-ray free electron lasers and third generation synchrotrons, but is of interest to any application in which microscopic samples must be delivered in a fully solvated state and with microscopic precision. By utilizing a gas dynamic virtual nozzle (GDVN) to generate a sample-containing liquid jet of diameter ranging from 300 nm to 20 µm, the injector avoids the clogging problems associated in this size range with conventional Rayleigh jets. A differential pumping system incorporated into the injector shields the experimental chamber from the gas load of the GDVN, making the injector compatible with high vacuum systems. The injector houses a fiber-optically coupled pump laser to illuminate the jet for pump-probe experiments and a hermetically sealed microscope to observe the liquid jet for diagnostics and alignment during operation. This injector system has now been used during several experimental runs at the Linac Coherent Light Source. Recent refinements in GDVN design are also presented.

Qualitative content analysis of online news media coverage of weight loss surgery and related reader comments.

The media has the ability to affect public opinion and policy direction. Prevalence of morbid obesity in Canada is increasing; as is the only effective long-term treatment, weight loss surgery (WLS). Limited research has explored media re/presentations of WLS. The purpose of this study was to examine national online news coverage (and reader comments) of WLS using content analysis. We sought to understand the dominant messages being conveyed within the news texts and reader comments, specifically whose voice was represented, who was the intended audience and what was the overall tone. Articles and comments were retrieved from the Canadian Broadcasting Corporation news web site and analysed using line-by-line techniques. Articles were predominantly 'positive/supportive' (63%) in tone and frequently presented the voices and opinions of 'experts' conveying a biomedical perspective. Comments were overwhelmingly 'negative' (56%) and often derogatory including such language as 'piggy' and 'fatty'. Comments were almost exclusively anonymous (99%) and were frequently directed at other commenters (33%) and 'fat' people (6%). The potentially problematic nature of media framing and reader comments, particularly as they could relate to weight-based stigmatization and discrimination is discussed.

New light on disordered ensembles: ab initio structure determination of one particle from scattering fluctuations of many copies.

We report on the first experimental ab initio reconstruction of an image of a single particle from fluctuations in the scattering from an ensemble of copies, randomly oriented about an axis. The method is applicable to identical particles frozen in space or time (as by snapshot diffraction from an x-ray free electron laser). These fluctuations enhance information obtainable from an experiment such as conventional small angle x-ray scattering.

Towards ETEM serial crystallography: Electron diffraction from liquid jets.

A sufficiently thin column of liquid was produced to permit penetration with a 200 keV electron beam as evidenced by the observation of diffraction rings due to the intermolecular spacing of the liquid samples. For liquid thickness below 800 nm, the diffraction rings became visible above the inelastic background. Studies were carried out in the environmental chamber of a transmission electron microscope using water and isopropanol.