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Upper GI endoscopy is speculated to be an aerosol-generating procedure (AGP). Robust evidence exists for aerosol transmission of severe acute respiratory syndrome coronavirus 2. The quality of data available confirming aerosol generation during GI endoscopy is limited. We aimed to objectively demonstrate that GI endoscopy is an AGP and illustrate the mechanism by which the greatest risk for aerosolization of droplets during endoscopy may occur.Aerosolized droplets generated during insertion and withdrawal of an endoscope and with passage of various tools through the endoscopic working channel using 2 experimental apparatuses modeling an upper GI tract (ie, a fluid-filled tube and a lamb esophagus) were qualitatively assessed by laser light scattering.Insertion and withdrawal of the upper endoscope into the upper GI tract models generated numerous aerosolized particles. A large number of brightly scattering particles were observed at the site of insertion and withdrawal of the endoscope. Passage of a cytology brush, biopsy forceps, and hemostatic clip through the working endoscope channel also generated aerosolized particles but in fewer numbers. There was no significant variation in quantity or brightness of droplets generated on testing different biopsy valve cap models or when suctioning fluid with an open versus closed biopsy valve cap. These results were reproducible over several trials.We illustrate in an objective manner that upper GI endoscopy is an AGP. These findings may have implications for transmission of infectious airborne pathogens outside of severe acute respiratory syndrome coronavirus 2 and can help to inform guidance on appropriate personal protective equipment use and other measures for transmission risk mitigation during GI endoscopy.

The Russian invasion of Ukraine on February 24, 2022, has displaced more than a quarter of the population. Assessing disease burdens among displaced people is instrumental in informing global public health and humanitarian aid efforts. We estimated the disease burden in Ukrainians displaced both within Ukraine and to other countries by combining a spatiotemporal model of forcible displacement with age- and gender-specific estimates of cardiovascular disease (CVD), diabetes, cancer, HIV, and tuberculosis (TB) in each of Ukraine’s 629 raions (i.e., districts). Among displaced Ukrainians as of May 13, we estimated that more than 2.63 million have CVDs, at least 615,000 have diabetes, and over 98,500 have cancer. In addition, more than 86,000 forcibly displaced individuals are living with HIV, and approximately 13,500 have TB. We estimated that the disease prevalence among refugees was lower than the national disease prevalence before the invasion. Accounting for internal displacement and healthcare facilities impacted by the conflict, we estimated that the number of people per hospital has increased by more than two-fold in some areas. As regional healthcare systems come under increasing strain, these estimates can inform the allocation of critical resources under shifting disease burdens.

Time-resolved small- and wide-angle X-ray scattering (SAXS/WAXS) studies of proteins in solution based on the pump-probe approach unveil structural information from intermediates over a broad range of length and time scales. In spite of the promise of this methodology, only a fraction of the wealth of information encoded in scattering data has been extracted in studies performed thus far. Here, we discuss the methodology, summarize results from recent time-resolved X-ray scattering studies, and examine the potential to extract additional information from these scattering curves.

The SARS‐CoV‐2 virus is highly contagious, as demonstrated by numerous well‐documented superspreading events. The infection commonly starts in the upper respiratory tract (URT) but can migrate to the lower respiratory tract (LRT) and other organs, often with severe consequences. Whereas LRT infection can lead to shedding of virus via breath and cough droplets, URT infection enables shedding via abundant speech droplets. Their viral load can be high in carriers with mild or no symptoms, an observation linked to the abundance of SARS‐CoV‐2‐susceptible cells in the oral cavity epithelium. Expelled droplets rapidly lose water through evaporation, with the smaller ones transforming into long‐lived aerosol. Although the largest speech droplets can carry more virions, they are few in number, fall to the ground rapidly and therefore play a relatively minor role in transmission. Of more concern is small speech aerosol, which can descend deep into the LRT and cause severe disease. However, since their total volume is small, the amount of virus they carry is low. Nevertheless, in closed environments with inadequate ventilation, they can accumulate, which elevates the risk of direct LRT infection. Of most concern is the large fraction of speech aerosol that is intermediate‐sized because it remains suspended in air for minutes and can be transported over considerable distances by convective air currents. The abundance of this speech‐generated aerosol, combined with its high viral load in pre‐ and asymptomatic individuals, strongly implicates airborne transmission of SARS‐CoV‐2 through speech as the primary contributor to its rapid spread.

Oral Fluid Droplets Generated during Speech In this experiment, investigators used a laser in a darkened box to visualize droplets that were generated during speech. The droplets decreased in numbe...

High efficiency of light harvesting in photosynthetic pigment–protein complexes is governed by evolutionary-perfected protein-assisted tuning of individual pigment properties and interpigment interactions. Due to the large number of spectrally overlapping pigments in a typical photosynthetic complex, experimental methods often fail to unambiguously identify individual chromophore properties. Here, we report a first-principles-based modeling protocol capable of predicting properties of pigments in protein environment to a high precision. The technique was applied to successfully uncover electronic properties of the Fenna–Matthews–Olson (FMO) pigment–protein complex. Each of the three subunits of the FMO complex contains eight strongly coupled bacteriochlorophyll a (BChl a) pigments. The excitonic structure of FMO can be described by an electronic Hamiltonian containing excitation (site) energies of BChl a pigments and electronic couplings between them. Several such Hamiltonians have been developed in the p...

During a pandemic, identifying modes of transmission is paramount to devise effective and practical mitigation strategies. Mohamed Abbas and Didier Pittet1Abbas M Pittet D Surfing the COVID-19 scientific wave.Lancet Infect Dis. 2020; (published online June 30.)https://doi.org/10.1016/S1473-3099(20)30558-2Summary Full Text Full Text PDF Scopus (0) Google Scholar challenge the conclusions of our reports that normal speaking might be an important mode of transmission for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), involving small particles that remain airborne for minutes.2Anfinrud P Stadnytskyi V Bax CE Bax A Visualizing speech-generated oral fluid droplets with laser light scattering.N Engl J Med. 2020; 382: 2061-2063Crossref PubMed Scopus (50) Google Scholar, 3Stadnytskyi V Bax CE Bax A Anfinrud P The airborne lifetime of small speech droplets and their potential importance in SARS-CoV-2 transmission.Proc Natl Acad Sci USA. 2020; 117: 11875-11877Crossref PubMed Scopus (41) Google Scholar Whereas the opening remarks in Abbas and Pittet's correspondence are irrelevant to our work, we eagerly welcome an intellectual debate on the scientific merits of our research. In their correspondence, they claim that our “findings have no immediate implications”.1Abbas M Pittet D Surfing the COVID-19 scientific wave.Lancet Infect Dis. 2020; (published online June 30.)https://doi.org/10.1016/S1473-3099(20)30558-2Summary Full Text Full Text PDF Scopus (0) Google Scholar Nothing could be further from the truth. While we refer readers to the appendix (pp 1–5) for a detailed response to all issues raised, we here address two of Abbas and Pittet's more pertinent concerns.

Speech droplets generated by asymptomatic carriers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are increasingly considered to be a likely mode of disease transmission. Highly sensitive laser light scattering observations have revealed that loud speech can emit thousands of oral fluid droplets per second. In a closed, stagnant air environment, they disappear from the window of view with time constants in the range of 8 to 14 min, which corresponds to droplet nuclei of ca. 4 μm diameter, or 12- to 21-μm droplets prior to dehydration. These observations confirm that there is a substantial probability that normal speaking causes airborne virus transmission in confined environments.

Speaking may be a primary mode of transmission of SARS-CoV-2. Considering that reports of asymptomatic transmission account for 50-80% of COVID-19 cases and that saliva has peak viral loads at time of patient presentation, droplet emission while speaking could be a significant factor driving transmission and warrants further study. We used a planar beam of laser light passing through a dust-free enclosure to detect saliva droplets emitted while speaking. We found that saying the words ‘Stay Healthy’ generates thousands of droplets that are otherwise invisible to the naked eye. A damp homemade cloth face mask dramatically reduced droplet excretion, with none of the spoken words causing a droplet rise above the background. Our preliminary findings have important implications for pandemic mitigation efforts.

In this paper we report the steady-state optical properties of a series of site-directed mutants in the Fenna-Matthews-Olson (FMO) complex of Chlorobaculum tepidum, a photosynthetic green sulfur bacterium. The FMO antenna complex has historically been used as a model system for energy transfer due to the water-soluble nature of the protein, its stability at room temperature, as well as the availability of high-resolution structural data. Eight FMO mutants were constructed with changes in the environment of each of the bacteriochlorophyll a pigments found within each monomer of the homotrimeric FMO complex. Our results reveal multiple changes in low temperature absorption, as well as room temperature CD in each mutant compared to the wild-type FMO complex. These datasets were subsequently used to model the site energies of each pigment in the FMO complex by employing three different Hamiltonians from the literature. This enabled a basic approximation of the site energy shifts imparted on each pigment by the changed amino acid residue. These simulations suggest that, while the three Hamiltonians used in this work provide good fits to the wild-type FMO absorption spectrum, further efforts are required to obtain good fits to the mutant minus wild-type absorption difference spectra. This demonstrates that the use of FMO mutants can be a valuable tool to refine and iterate the current models of energy transfer in this system.

The 2.5 A structure of the cytochrome (cyt) b6 f complex provides a basis for control of the rate-limiting electron transfer step of oxygenic photosynthesis associated with the plastoquinol/quinone exchange pathway. Here, a structural change was made at a site containing two proline residues which border the intra-cyt pathway for plastoquinol/quinone exchange. The proline side chains confer a larger aperture for passage of plastoquinol/quinone. Change of these prolines to alanine in the cyanobacterium Synechococcus sp. PCC 7002 results in attenuation of this rate-limiting step, observed by a two-fold reduction in the rate of cell growth, O2 evolution, and plastoquinol-mediated reduction of cyt f. This study demonstrates modification by site-directed mutagenesis of photosynthetic energy transduction based on rational application of information in the atomic structure.

It is well-known that tetrameric hemoglobin binds ligands cooperatively by undergoing a ligand-induced T → R quaternary structure transition, a structure-function relationship that has long served as a model system for understanding allostery in proteins. However, kinetic studies of the reverse, R → T quaternary structure transition following photolysis of carbonmonoxyhemoglobin (HbCO) reveal complex behavior that may be better explained by the presence of two different R quaternary structures coexisting in thermal equilibrium. Indeed, we report here time-resolved small- and wide-angle X-ray scattering (SAXS/WAXS) patterns of HbCO following a temperature jump that not only provide unambiguous evidence for more than one R state, but also unveil the time scale for interconversion between them. Since the time scale for the photolysis-induced R → T transition is likely different for different R-states, this structural heterogeneity must be accounted for to properly explain the kinetic heterogeneity observed in time-resolved spectroscopic studies following photolysis of HbCO.

We describe an optical near shot-noise limited time-resolved circular dichroism (TRCD) pump-probe spectrometer capable of reliably measuring circular dichroism signals in the order of μdeg with nanosecond time resolution. Such sensitivity is achieved through a modification of existing TRCD designs and introduction of a new data processing protocol that eliminates approximations that have caused substantial nonlinearities in past measurements and allows the measurement of absorption and circular dichroism transients simultaneously with a single pump pulse. The exceptional signal-to-noise ratio of the described setup makes the TRCD technique applicable to a large range of non-biological and biological systems. The spectrometer was used to record, for the first time, weak TRCD kinetics associated with the triplet state energy transfer in the photosynthetic Fenna-Matthews-Olson antenna pigment-protein complex.

The intramembrane cytochrome bc1 complex of the photosynthetic bacterium Rhodobacter capsulatus and the cytochrome b6f complex, which functions in oxygenic photosynthesis, utilize two pairs of b-hemes in a symmetric dimer to accomplish proton-coupled electron transfer. The transmembrane electron transfer pathway in each complex was identified through the novel use of heme Soret band excitonic circular dichroism (CD) spectra, for which the responsible heme–heme interactions were determined from crystal structures. Kinetics of heme reduction and CD amplitude change were measured simultaneously. For bc1, in which the redox potentials of the transmembrane heme pair are separated by 160 mV, heme reduction occurs preferentially to the higher-potential intermonomer heme pair on the electronegative (n) side of the complex. This contrasts with the b6f complex, where the redox potential difference between transmembrane intramonomer p- and n-side hemes is substantially smaller and the n–p pair is preferentially redu...

The cytochrome bc1 complex utilizes two pairs of b-hemes in a mostly symmetric dimer to accomplish trans-membrane electron transfer and generation of a trans-membrane proton electrochemical potential gradient. Analysis of the structure of the complex in the purple photosynthetic bacterium, Rhodobacter capsulatus, allowed determination of the identity of the strongly interacting heme pair. Determination of the preferred heme sites of electron residence utilized Soret band excitonic circular dichroism (CD) spectra to assay heme−heme interactions. Simultaneous kinetics of heme reduction and generation of the CD spectra imply that the thermodynamically favorable pair-wise heme occupancy utilizes the intra-monomer b-heme pair that exhibits different heme redox potentials on the electrochemically positive and negative sides of the cytochrome complex. This contradicts the expectation, based on the assumption of a dielectrically homogeneous protein environment, that the two higher potential hemes on the electronegative cytoplasmic side of the complex would be preferentially reduced. Relative to the protein medium in the structure that separates the two higher potential hemes on the electronegative side of the complex, a relatively large dielectric constant must exist between the intra-monomer b-hemes on the two sides of the complex, thus allowing increased polarizabilty and a smaller electrostatic repulsion between the reduced hemes. Thus, the distribution of dielectric constants in the core of the intra-membrane cytochrome complex, which mediate inter-heme electrostatic interactions, and which are a measure of local hydrophobicity, is heterogeneous.

The cytochrome b6f complex, a member of the cytochrome bc family that mediates energy transduction in photosynthetic and respiratory membranes, is a hetero-oligomeric complex that utilizes two pairs of b-hemes in a symmetric dimer to accomplish trans-membrane electron transfer, quinone oxidation–reduction, and generation of a proton electrochemical potential. Analysis of electron storage in this pathway, utilizing simultaneous measurement of heme reduction, and of circular dichroism (CD) spectra, to assay heme–heme interactions, implies a heterogeneous distribution of the dielectric constants that mediate electrostatic interactions between the four hemes in the complex. Crystallographic information was used to determine the identity of the interacting hemes. The Soret band CD signal is dominated by excitonic interaction between the intramonomer b-hemes, bn and bp, on the electrochemically negative and positive sides of the complex. Kinetic data imply that the most probable pathway for transfer of the two electrons needed for quinone oxidation–reduction utilizes this intramonomer heme pair, contradicting the expectation based on heme redox potentials and thermodynamics, that the two higher potential hemes bn on different monomers would be preferentially reduced. Energetically preferred intramonomer electron storage of electrons on the intramonomer b-hemes is found to require heterogeneity of interheme dielectric constants. Relative to the medium separating the two higher potential hemes bn, a relatively large dielectric constant must exist between the intramonomer b-hemes, allowing a smaller electrostatic repulsion between the reduced hemes. Heterogeneity of dielectric constants is an additional structure–function parameter of membrane protein complexes.

Laser-induced incandescence (LII) of carbon microparticles in epoxy matrixes is studied. Non-monotonical behaviour of LII from doze of laser irradiation is observed. The proposed model interprets the majority of the observed experimental data.