Structure of the Angiotensin receptor revealed by serial femtosecond crystallography.

Angiotensin II type 1 receptor (AT(1)R) is a G protein-coupled receptor that serves as a primary regulator for blood pressure maintenance. Although several anti-hypertensive drugs have been developed as AT(1)R blockers (ARBs), the structural basis for AT(1)R ligand-binding and regulation has remained elusive, mostly due to the difficulties of growing high-quality crystals for structure determination using synchrotron radiation. By applying the recently developed method of serial femtosecond crystallography at an X-ray free-electron laser, we successfully determined the room-temperature crystal structure of the human AT(1)R in complex with its selective antagonist ZD7155 at 2.9-Å resolution. The AT(1)R-ZD7155 complex structure revealed key structural features of AT(1)R and critical interactions for ZD7155 binding. Docking simulations of the clinically used ARBs into the AT(1)R structure further elucidated both the common and distinct binding modes for these anti-hypertensive drugs. Our results thereby provide fundamental insights into AT(1)R structure-function relationship and structure-based drug design.

Predicting cancer-specific vulnerability via data-driven detection of synthetic lethality.

Synthetic lethality occurs when the inhibition of two genes is lethal while the inhibition of each single gene is not. It can be harnessed to selectively treat cancer by identifying inactive genes in a given cancer and targeting their synthetic lethal (SL) partners. We present a data-driven computational pipeline for the genome-wide identification of SL interactions in cancer by analyzing large volumes of cancer genomic data. First, we show that the approach successfully captures known SL partners of tumor suppressors and oncogenes. We then validate SL predictions obtained for the tumor suppressor VHL. Next, we construct a genome-wide network of SL interactions in cancer and demonstrate its value in predicting gene essentiality and clinical prognosis. Finally, we identify synthetic lethality arising from gene overactivation and use it to predict drug efficacy. These results form a computational basis for exploiting synthetic lethality to uncover cancer-specific susceptibilities.

Ultrafast structural changes direct the first molecular events of vision.

Vision is initiated by the rhodopsin family of light-sensitive G protein-coupled receptors (GPCRs)1. A photon is absorbed by the 11-cis retinal chromophore of rhodopsin, which isomerizes within 200 femtoseconds to the all-trans conformation2, thereby initiating the cellular signal transduction processes that ultimately lead to vision. However, the intramolecular mechanism by which the photoactivated retinal induces the activation events inside rhodopsin remains experimentally unclear. Here we use ultrafast time-resolved crystallography at room temperature3 to determine how an isomerized twisted all-trans retinal stores the photon energy that is required to initiate the protein conformational changes associated with the formation of the G protein-binding signalling state. The distorted retinal at a 1-ps time delay after photoactivation has pulled away from half of its numerous interactions with its binding pocket, and the excess of the photon energy is released through an anisotropic protein breathing motion in the direction of the extracellular space. Notably, the very early structural motions in the protein side chains of rhodopsin appear in regions that are involved in later stages of the conserved class A GPCR activation mechanism. Our study sheds light on the earliest stages of vision in vertebrates and points to fundamental aspects of the molecular mechanisms of agonist-mediated GPCR activation.

Femtosecond-to-millisecond structural changes in a light-driven sodium pump.

Light-driven sodium pumps actively transport small cations across cellular membranes1. These pumps are used by microorganisms to convert light into membrane potential and have become useful optogenetic tools with applications in neuroscience. Although the resting state structures of the prototypical sodium pump Krokinobacter eikastus rhodopsin 2 (KR2) have been solved2,3, it is unclear how structural alterations over time allow sodium to be translocated against a concentration gradient. Here, using the Swiss X-ray Free Electron Laser4, we have collected serial crystallographic data at ten pump-probe delays from femtoseconds to milliseconds. High-resolution structural snapshots throughout the KR2 photocycle show how retinal isomerization is completed on the femtosecond timescale and changes the local structure of the binding pocket in the early nanoseconds. Subsequent rearrangements and deprotonation of the retinal Schiff base open an electrostatic gate in microseconds. Structural and spectroscopic data, in combination with quantum chemical calculations, indicate that a sodium ion binds transiently close to the retinal within one millisecond. In the last structural intermediate, at 20 milliseconds after activation, we identified a potential second sodium-binding site close to the extracellular exit. These results provide direct molecular insight into the dynamics of active cation transport across biological membranes.

Erratum: Spontaneous Domain Formation in Spherically Confined Elastic Filaments [Phys. Rev. Lett. 123, 047801 (2019)].

This corrects the article DOI: 10.1103/PhysRevLett.123.047801.

Do high-protein diets have the potential to reduce gut barrier function in a sex-dependent manner?

PURPOSE: Impaired gut barrier function is associated with systemic inflammation and many chronic diseases. Undigested dietary proteins are fermented in the colon by the gut microbiota which produces nitrogenous metabolites shown to reduce barrier function in vitro. With growing evidence of sex-based differences in gut microbiotas, we determined whether there were sex by dietary protein interactions which could differentially impact barrier function via microbiota modification. METHODS: Fermentation systems were inoculated with faeces from healthy males (n = 5) and females (n = 5) and supplemented with 0.9 g of non-hydrolysed proteins sourced from whey, fish, milk, soya, egg, pea, or mycoprotein. Microbial populations were quantified using fluorescence in situ hybridisation with flow cytometry. Metabolite concentrations were analysed using gas chromatography, solid phase microextraction coupled with gas chromatography-mass spectrometry and ELISA. RESULTS: Increased protein availability resulted in increased proteolytic Bacteroides spp (p < 0.01) and Clostridium coccoides (p < 0.01), along with increased phenol (p < 0.01), p-cresol (p < 0.01), indole (p = 0.018) and ammonia (p < 0.01), varying by protein type. Counts of Clostridium cluster IX (p = 0.03) and concentration of p-cresol (p = 0.025) increased in males, while females produced more ammonia (p = 0.02), irrespective of protein type. Further, we observed significant sex-protein interactions affecting bacterial populations and metabolites (p < 0.005). CONCLUSIONS: Our findings suggest that protein fermentation by the gut microbiota in vitro is influenced by both protein source and the donor's sex. Should these results be confirmed through human studies, they could have major implications for developing dietary recommendations tailored by sex to prevent chronic illnesses.

Report on progress in physics: observation of the Breit-Wheeler process and vacuum birefringence in heavy-ion collisions.

This report reviews the effort over several decades to observe the linear Breit-Wheeler process (γγ→e+e-) and vacuum birefringence (VB) in high-energy particle and heavy-ion collider experiment. This report, motivated by the STAR collaboration's recent observations, attempts to summarize the key issues related to the interpretation of polarizedγγ→l+l-measurements in high-energy experiments. To that end, we start by reviewing the historical context and essential theoretical developments, before focusing on the decades of progress made in high-energy collider experiments. Special attention is given to the evolution in experimental approaches in response to various challenges, to the demanding detector capabilities required to unambiguously identify the linear Breit-Wheeler process, and to the connections with VB. We close the report with a discussion, followed by a look at near-future opportunities for utilizing these discoveries and for testing quantum electrodynamics in previously unexplored regimes.

Managing and Preventing Migraine in the Emergency Department: A Review.

Migraine is a leading cause of disability worldwide, and acute migraine attacks are a common reason for patients to seek care in the emergency department (ED). There have been recent advancements in the care of patients with migraine, specifically emerging evidence for nerve blocks and new pharmacological classes of medications like gepants and ditans. This article serves as a comprehensive review of migraine in the ED, including diagnosis and management of acute complications of migraine (eg, status migrainosus, migrainous infarct, persistent aura without infarction, and aura-triggered seizure) and use of evidence-based migraine-specific treatments in the ED. It highlights the role of migraine preventive medications and provides a framework for emergency physicians to prescribe them to eligible patients. Finally, it evaluates the evidence for nerve blocks in the treatment of migraine and introduces the possible role of gepants and ditans in the care of patients with migraine in the ED.

The peripheral dopamine 2 receptor antagonist domperidone attenuates ethanol enhancement of dopamine levels in the nucleus accumbens.

BACKGROUND: Dopamine neuron firing in the ventral tegmental area (VTA) and dopamine release in the nucleus accumbens have been implicated in reward learning. Ethanol is known to increase both dopamine neuron firing in the VTA and dopamine levels in the nucleus accumbens. Despite this, some discrepancies exist between the dose of ethanol required to enhance firing in vivo and ex vivo. In the present study we investigated the effects of peripheral dopamine 2 subtype receptor antagonism on ethanol's effects on dopamine neurotransmission. METHODS: Plasma catecholamine levels were assessed following ethanol administration across four different doses of EtOH. Microdialysis and voltammetry were used to assess the effects of domperidone pretreatment on ethanol-mediated increases in dopamine release in the nucleus accumbens. A place conditioning paradigm was used to assess conditioned preference for ethanol and whether domperidone pretreatment altered this preference. Open-field and loss-of-righting reflex paradigms were used to assess the effects of domperidone on ethanol-induced sedation. A rotarod apparatus was used to assess the effects of domperidone on ethanol-induced motor impairment. RESULTS: Domperidone attenuated ethanol's enhancement of mesolimbic dopamine release under non-physiological conditions at intermediate (1.0 and 2.0 g/kg) doses of ethanol. Domperidone also decreased EtOH-induced sedation at 2.0 g/kg. Domperidone did not alter ethanol conditioned place preference nor did it affect ethanol-induced motor impairment. CONCLUSIONS: These results show that peripheral dopamine 2 receptors mediate some of the effects of ethanol on nonphysiological dopamine neurotransmission, although these effects are not related to the rewarding properties of ethanol.

Current practice for primary headache disorders and perspectives on peripheral nerve blocks among emergency physicians in Canada: A national survey.

OBJECTIVE: This national postal survey aimed to examine Canadian emergency physicians' practice patterns with respect to drug treatment and perspectives on peripheral nerve blocks. BACKGROUND: The treatment of primary headache disorders in the emergency department is variable. METHODS: We surveyed 500 emergency physicians listed in the Canadian Medical Directory according to a modified Dillman's method: an initial invitation was followed by up to four reminders to nonresponders. Physicians were asked questions regarding their frequency of medication administration and perspectives toward peripheral nerve blocks. RESULTS: Of 500 mailed surveys, 468 were delivered and 179 physicians responded (response rate = 38.2%). The majority of physicians were men (92/144, 63.9%); 80.6% (116/144) had been in practice for greater than or equal to 10 years with 50.7% (75/148) in a community or district general teaching hospital. Commonly used pharmacotherapies for primary headaches were intravenous dopamine receptor antagonists (69%), co-administration of ketorolac and a dopamine receptor antagonist (54.2%), intravenous fluid boluses (54%), nonsteroidal anti-inflammatory drugs (NSAIDs) alone (53.5%), and acetaminophen (51.4%). Only 80 of 144 physicians (55.6%) reported previous experience with peripheral nerve blocks (95% confidence interval [CI] = 48%-65%). The majority (68/80, 85.0%) agreed peripheral nerve blocks are safe and 55.1% (43/78) agreed they are effective. The vast majority (118/140, 84.3%) would consider peripheral nerve blocks as a first-line treatment option given sufficient evidence from a future trial (95% CI = 78%-90%). CONCLUSION: NSAIDs alone, as well as dopamine receptor antagonists with or without ketorolac are commonly used for primary headache in Canadian emergency departments. A large proportion of physicians have never used a peripheral nerve block in their practice; among those who have experience with peripheral nerve blocks, the majority find them safe and effective. The majority of respondents would consider peripheral nerve blocks as a first-line treatment option given sufficient evidence from a future trial.

Macromolecular diffractive imaging using imperfect crystals.

The three-dimensional structures of macromolecules and their complexes are mainly elucidated by X-ray protein crystallography. A major limitation of this method is access to high-quality crystals, which is necessary to ensure X-ray diffraction extends to sufficiently large scattering angles and hence yields information of sufficiently high resolution with which to solve the crystal structure. The observation that crystals with reduced unit-cell volumes and tighter macromolecular packing often produce higher-resolution Bragg peaks suggests that crystallographic resolution for some macromolecules may be limited not by their heterogeneity, but by a deviation of strict positional ordering of the crystalline lattice. Such displacements of molecules from the ideal lattice give rise to a continuous diffraction pattern that is equal to the incoherent sum of diffraction from rigid individual molecular complexes aligned along several discrete crystallographic orientations and that, consequently, contains more information than Bragg peaks alone. Although such continuous diffraction patterns have long been observed--and are of interest as a source of information about the dynamics of proteins--they have not been used for structure determination. Here we show for crystals of the integral membrane protein complex photosystem II that lattice disorder increases the information content and the resolution of the diffraction pattern well beyond the 4.5-ångström limit of measurable Bragg peaks, which allows us to phase the pattern directly. Using the molecular envelope conventionally determined at 4.5 ångströms as a constraint, we obtain a static image of the photosystem II dimer at a resolution of 3.5 ångströms. This result shows that continuous diffraction can be used to overcome what have long been supposed to be the resolution limits of macromolecular crystallography, using a method that exploits commonly encountered imperfect crystals and enables model-free phasing.

Quantum kinematics in terms of observable quantities and the chirality of entangled two-qubit states.

We consider the kinematics of bipartite quantum states as determined by observable quantities, in particular the Bloch vectors of the subsystems. In examining the simplest case of a pair of two-level systems, there is a remarkable connection between the presence of non-classical correlations and the chirality of the two bases generated by the singular value decomposition of the correlation matrix of the Bloch vectors. We investigate the limits imposed by quantum mechanics of this effect and its relationship with other methods on quantifying the system's non-classical behavior.

Experimental evaluation of respiratory droplet spread to rooms connected by a central ventilation system.

This article presents results from an experimental study to ascertain the transmissibility of the SARS-CoV-2 virus between rooms in a building that are connected by a central ventilation system. Respiratory droplet surrogates made of mucus and virus mimics were released in one room in a test building, and measurements of concentration levels were made in other rooms connected via the ventilation system. The paper presents experimental results for different ventilation system configurations, including ventilation rate, filtration level (up to MERV-13), and fractional outdoor air intake. The most important finding is that respiratory droplets can and do transit through central ventilation systems, suggesting a mechanism for viral transmission (and COVID-19 specifically) within the built environment in reasonable agreement with well-mixed models. We also find the deposition of small droplets (0.5-4 µm) on room walls to be negligibly small.

A Systematic Review of the Cost of Chronic Pelvic Pain in Women.

OBJECTIVE: To systematically summarize the evidence on costs related to chronic pelvic pain (CPP) for women. DATA SOURCES: Electronic databases (MEDLINE, EMBASE, PubMed, and Cochrane Library) were searched for English and French articles published from 1990 to January 2021 STUDY SELECTION: Of 1304 articles screened, 67 were screened in full-text form, and a total of 13 articles were included in the final analysis. Articles included involved cost studies that estimated hospital or health system costs for pelvic pain, dysmenorrhea, dyspareunia, endometriosis with pain, interstitial cystitis, or painful bladder syndrome. DATA EXTRACTION AND SYNTHESIS: A standardized form was created to extract study setting, design, and population; patient demographics; study duration; and reported costs of CPP components and amounts. Two independent reviewers completed the data extraction, and discrepancies were resolved through discussion with a third reviewer. CONCLUSION: Estimated health care costs ranged from US$1367 to US$7043 per woman per year. Prescription costs ranged from US$193 to US$2457 per woman per year. Indirect costs ranged from US$4216 to US$12 789 per woman per year. Combined costs ranged from US$1820 to US$20 898 per woman per year. The yearly costs of CPP varied according to country; yearly costs were estimated to be $2.8 billion, ¥191,680 to ¥246,488, and $16 970 to $20 898 per woman per year in the United Sates, Japan, and Australia, respectively. The literature suggests that CPP represents a considerable economic burden on women and health care systems internationally, with indirect costs contributing a significant portion of total costs.

Translational Applications of Wearable Sensors in Education: Implementation and Efficacy.

BACKGROUND: Adding new approaches to teaching curriculums can be both expensive and complex to learn. The aim of this research was to gain insight into students' literacy and confidence in learning sports science with new wearable technologies, specifically a novel program known as STEMfit. METHODS: A three-phase design was carried out, with 36 students participating and exposed to wearable devices and associated software. This was to determine whether the technology hardware (phase one) and associated software (phase two) were used in a positive way that demonstrated user confidence. RESULTS: Hardware included choosing a scalable wearable device that worked in conjunction with familiar and readily available software (Microsoft Excel) that extracted data through VBA coding. This allowed for students to experience and provide survey feedback on the usability and confidence gained when interacting with the STEMfit program. Outcomes indicated strong acceptance of the program, with high levels of motivation, resulting in a positive uptake of wearable technology as a teaching tool by students. The initial finding of this study offers an opportunity to further test the STEMfit program on other student cohorts as well as testing the scalability of the system into other year groups at the university level.

Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser.

G-protein-coupled receptors (GPCRs) signal primarily through G proteins or arrestins. Arrestin binding to GPCRs blocks G protein interaction and redirects signalling to numerous G-protein-independent pathways. Here we report the crystal structure of a constitutively active form of human rhodopsin bound to a pre-activated form of the mouse visual arrestin, determined by serial femtosecond X-ray laser crystallography. Together with extensive biochemical and mutagenesis data, the structure reveals an overall architecture of the rhodopsin-arrestin assembly in which rhodopsin uses distinct structural elements, including transmembrane helix 7 and helix 8, to recruit arrestin. Correspondingly, arrestin adopts the pre-activated conformation, with a ∼20° rotation between the amino and carboxy domains, which opens up a cleft in arrestin to accommodate a short helix formed by the second intracellular loop of rhodopsin. This structure provides a basis for understanding GPCR-mediated arrestin-biased signalling and demonstrates the power of X-ray lasers for advancing the frontiers of structural biology.

Limitations of Hartree-Fock with quantum resources.

The Hartree-Fock problem provides the conceptual and mathematical underpinning of a large portion of quantum chemistry. As efforts in quantum technology aim to enhance computational chemistry algorithms, the Hartree-Fock method, central to many other numerical approaches, is a natural target for quantum enhanced algorithms. While quantum computers and quantum simulation offer many prospects for the future of modern chemistry, the non-deterministic polynomial-complete Hartree-Fock problem is not a likely candidate. We highlight this fact from a number of perspectives including computational complexity, practical examples, and the full characterization of energy landscapes for simple systems.

The Effect of Cleat Position on Running Using Acceleration-Derived Data in the Context of Triathlons.

Appropriate cycling cleat adjustment could improve triathlon performance in both cycling and running. Prior recommendations regarding cleat adjustment have comprised aligning the first metatarsal head above the pedal spindle or somewhat forward. However, contemporary research has questioned this approach in triathlons due to the need to run immediately after cycling. Subsequently, moving the pedal cleat posteriorly could be more appropriate. This study evaluated the effectiveness of a triaxial accelerometer to determine acceleration magnitudes of the trunk in outdoor cycling in two different bicycle cleat positions and the consequential impact on trunk acceleration during running. Seven recreational triathletes performed a 20 km cycle and a 5 km run using their own triathlon bicycle complete with aerodynamic bars and gearing. Interpretation of data was evaluated based on cadence changes whilst triathletes cycled in an aerodynamic position in two cleat positions immediately followed by a self-paced overground run. The evaluation of accelerometer-derived data within a characteristic overground setting suggests a significant increase in total trunk acceleration magnitude during cycling with a posterior cleat with significant increases to longitudinal acceleration (p = 0.04) despite a small effect (d = 0.2) to the ratings of perceived exertion (RPE). Cycling with a posterior cleat significantly reduced longitudinal trunk acceleration in running and overall acceleration magnitudes (p < 0.0001) with a large effect size (d = 0.9) and a significant reduction in RPE (p = 0.02). In addition, running after cycling in a posterior cleat was faster compared to running after cycling in a standard cleat location. Practically, the magnitude of trunk acceleration during cycling in a posterior cleat position as well as running after posterior cleat cycling differed from that when cycling in the fore-aft position followed by running. Therefore, the notion that running varies after cycling is not merely an individual athlete's perception, but a valid observation that can be modified when cleat position is altered. Training specifically with a posterior cleat in cycling might improve running performance when trunk accelerations are analysed.

Evaluation of Accelerometer-Derived Data in the Context of Cycling Cadence and Saddle Height Changes in Triathlon.

In the multisport of triathlon cycling is the longest of the three sequential disciplines. Triathlon bicycles differ from road bicycles with steeper seat tube angles with a change to saddle height altering the seat tube angle. This study evaluated the effectiveness of a tri axial accelerometer to determine acceleration magnitudes of the trunk in outdoor cycling in two saddle positions. Interpretation of data was evaluated based on cadence changes whilst triathletes cycled in an aerodynamic position in two saddle positions. The evaluation of accelerometer derived data within a characteristic overground setting suggests a significant reduction in mediolateral acceleration of the trunk, yielding a 25.1% decrease when saddle height was altered alongside reduced rate of perceived exertion (3.9%). Minimal differences were observed in anteroposterior and longitudinal acceleration. Evaluation of sensor data revealed a polynomial expression of the subtle changes between both saddle positions. This study shows that a triaxial accelerometer has capability to continuously measure acceleration magnitude of trunk movements during an in-the-field, varied cadence cycle protocol. Accessible and practical sensor technology could be relevant for postural considerations when exploring saddle position in dynamic settings.

Investigation of potential aerosol transmission and infectivity of SARS-CoV-2 through central ventilation systems.

The COVID-19 pandemic has raised concern of viral spread within buildings. Although near-field transmission and infectious spread within individual rooms are well studied, the impact of aerosolized spread of SARS-CoV-2 via air handling systems within multiroom buildings remains unexplored. This study evaluates the concentrations and probabilities of infection for both building interior and exterior exposure sources using a well-mixed model in a multiroom building served by a central air handling system (without packaged terminal air conditioning). In particular, we compare the influence of filtration, air change rates, and the fraction of outdoor air. When the air supplied to the rooms comprises both outdoor air and recirculated air, we find filtration lowers the concentration and probability of infection the most in connected rooms. We find that increasing the air change rate removes virus from the source room faster but also increases the rate of exposure in connected rooms. Therefore, slower air change rates reduce infectivity in connected rooms at shorter durations. We further find that increasing the fraction of virus-free outdoor air is helpful, unless outdoor air is infective in which case pathogen exposure inside persists for hours after a short-term release. Increasing the outdoor air to 33% or the filter to MERV-13 decreases the infectivity in the connected rooms by 19% or 93% respectively, relative to a MERV-8 filter with 9% outdoor air based on 100 quanta/h of 5 µm droplets, a breathing rate of 0.48 m3/h, and the building dimensions and air handling system considered.