Atractylodes macrocephala Koidz (PAMK) has antioxidation and immunomodulatory effects. Therefore, we used CTX to construct an immunosuppression model for goslings to explore the mechanism by which PAMK alleviates cyclophosphamide (CTX)-induced ferroptosis in thymocytes and to provide a basis for a more profound elucidation of the immunomodulatory mechanism of PAMK. It was found that PAMK had a significant alleviating effect on CTX-induced thymus damage and thymocyte ferroptosis in goslings. Therefore, PAMK can be used as a natural alternative to antibiotics as a feed additive for immunomodulatory effects on goslings.During the brooding stage, the goslings are susceptible to various stresses, damaging the thymus and decreasing immune function. Ferroptosis is cell death caused by iron-ion-dependent lipid peroxidation, which has been reported to be closely related to organismal immunity. The polysaccharide of p < 0.05) and inhibited the CTX-induced activation of the ferroptosis pathway (p < 0.05). Conclusively, PAMK could reduce thymus injury by alleviating CTX-induced thymocyte ferroptosis in gosling to alleviate the immunosuppression caused by CTX in the organism.The present study aimed to explore the mechanism by which PAMK alleviates cyclophosphamide (CTX)-induced ferroptosis in thymocytes. One-day-old goslings were divided into four groups (10 goslings/group). The CON and CTX groups were fed a basic diet. The PAMK and CTX + PAMK groups were fed the basic diet mixed with PAMK (400 mg/kg). Moreover, the CTX and CTX + PAMK groups were given a daily injection of 40 mg/kg BW of CTX . On the other hand, the CON and PAMK groups were given 0.5 mL of sterilized saline into the leg muscle . The goslings were fed for 28 days. The ferroptosis pathway was enriched in transcriptome sequencing. Compared to the CON group, the thymus in the CTX group underwent injury, and the mitochondria of thymocytes showed features of ferroptosis. PAMK treatment alleviated ferroptosis in thymocytes and thymus injury, and CTX-induced elevated levels of oxidative stress and iron content restored GPX4 protein expression ( Ferroptosis is cell death caused by oxidative damage from iron-ion-dependent lipid peroxidation, leading to iron metabolic dysfunction and massive lipid peroxidation . FerroptThe thymus, as a central immune organ for T cell development and differentiation, has a significant regulatory role in the immune function of the organism . During Atractylodes macrocephala, which has been recorded in the Chinese medical classic Shengnong’s herba. Studies have shown that PAMK can promote the development of immune organs, regulate the proliferation and differentiation of immune cells, positively regulate the immune response capacity of the body, and alleviate the immunosuppression and immune organ damage caused by harmful stimuli, maintaining normal immune organ function and the homeostasis of the immune system [PAMK is the main active component of the Chinese medicine e system ,12. Our e system ,14, and e system .In this study, we constructed a model of immunosuppressed goslings using CTX, added PAMK for treatment. To explore the mechanism by which PAMK alleviates CTX-induced thymus injury, we first conducted transcriptome sequencing of the thymus and enriched the ferroptosis pathway. Based on transcriptome sequencing, we speculated that PAMK could alleviate CTX-induced ferroptosis in the thymocytes of goslings. Therefore, we observed the thymus histology and ultramicroscopic morphology and detected ferroptosis marker GPX4 and thymus iron content, as well as oxidative stress levels and genes related to the ferroptosis pathway. The mechanism of PAMK alleviating CTX-induced ferroptosis in thymocytes was preliminarily explored to provide a basis for more in-depth elucidation of the immunomodulatory mechanism of PAMK and a theoretical basis for the further development of natural plant additives.All goslings were purchased from Guangdong QingyuanJinyufeng Goose Co., Ltd., Qingyuan, China. Forty 1-day-old Magang goslings  were randomly divided into four groups of 10 goslings each and pre-fed for 3 days. The CON and CTX groups were fed a basic diet. The PAMK and CTX + PAMK groups were fed the basic diet mixed with PAMK (400 mg/kg). At 19, 20, and 21 days of age, the CTX and CTX + PAMK groups were given a daily injection of 40 mg/kg BW of CTX  into the leg muscle. On the other hand, the CON and PAMK groups were given 0.5 mL of sterilized saline into the leg muscle . The dosp-value < 0.05. All DEGs were visualized as heat maps and volcano maps using R packages: pheatmap and ggplot2, respectively. Gene ontology (GO) enrichment analysis, including biological processes, cellular components, and molecular functions, were performed using Blast2GO software to determine the biological roles of the DEGs.Transcriptome sequencing was performed by BGI Genomics Co., Ltd., . Total RNA was extracted from the thymus using TRIzol reagent . High-quality RNA was used to construct sequencing libraries and analyze the enrichment of differentially expressed genes (DEGs). First, data were filtered and assembled using HISAT2 software, and the expression of genes and transcripts were calculated using RSEM software. The interception criteria for DEGs were |logFC| ≥ 1 and adjusted −∆∆ct relative quantification method.Total RNA was extracted from the thymus of goslings using TRIzol reagent according to the manufacturer’s reagent instructions  and reverse-transcribed using reverse-transcription reagent . Subsequently, cDNA templates from the samples were amplified with SYBR Green . Primer sequences were designed according to the NCBI database . We usedWe collected thymus for protein extraction using RIPA lysis buffer  supplemented with protease inhibitors . The protein from the thymus was separated by electrophoresis. Transfer the protein to the PVDF membrane at 4 °C. After blocking with 5% skimmed milk at room temperature for 2 h, the membranes were incubated with GPX4 primary antibody  and GAPDH primary antibody  at 4 °C overnight. After incubation with peroxidase-conjugated secondary antibodies  for 1 h at room temperature. The signal was detected on an ImageQuant LAS 500  using the ECL Kit . GAPDH expression served as a loading control for quantification.We cut the paraffin-fixed blocks into consecutive coronal sections 5–6 μm thick. For routine histological examination, paraffin sections were stained with HE. The sections were scanned with a section scanner , and the HE-stained scanned sections were analyzed with CaseViewer. We observed changes in HE-stained sections of the thymus, selected three sections from each group with 10 randomly selected fields of view (200×), and measured the thickness of the thymic cortex area in each field of view for statistical analysis.Each thymus was divided into 1 mm three pieces and then fixed with 2.5% glutaraldehyde at 4 °C. Ultrathin sections of 50–70 nm thickness were prepared and stained with uranyl acetate  and lead citrate . Samples were observed with a transmission electron microscope  and with a scanning electron microscope . The ratio of dead cells to live cells was calculated at 3000×.Thymus sections were dewaxed in xylene and hydrated with a gradient ethanol solution. Antigen repair was performed on the sections using EDTA antigen repair buffer . Endogenous peroxidase was blocked with a 3% hydrogen peroxide solution. The thymus sections were blocked in BSA  for 1 h and and incubated with the GPX4 primary antibody  at 4 °C overnight. The secondary antibody was used at a 1/1000 dilution for 1 h at room temperature. DAPI was used to stain the cell nuclei. Images were acquired by a fluorescent inverted microscope . Anti-fluorescence quenching mounting tablets  were used for mounting. Image J was used to calculate the GPX4 fluorescence.6 cells/mL, mix well, and incubate for 15 min away from light. The cells were washed with PBS, centrifuged at 1600 rpm for 5 min, discarded the supernatant, and resuspended in 300 μL PBS. The resuspended cells were detected using a flowmeter .The content of fresh thymus intracellular ROS was measured using the ROS detection kit . Fresh thymus from goslings was collected and ground on a 70 μm cell sieve to filter the cells and into PBS. Centrifugation was performed at 1600 rpm for 5 min, and the supernatant was discarded. Add 200 μL DCFH-DA staining solution, adjust the concentration of cells to 5 × 10GSH (A006-2-1), MDA (A003-1-2), and tissue Fe (A039-2-1) level assays were performed using the kit  according to the manufacturer’s instructions.p < 0.05 were considered as statistically significant.The experimental data were analyzed by one-way ANOVA using GraphPad Prism 7.0. The differences between the four groups were compared by Tukey’s multiple comparison method, and To clarify the mechanism of action of CTX on thymus injury and PAMK on thymus protection, we first performed RNA sequencing of CTX and the thymus of CTX and PAMK co-treated goslings to systematically investigate the biological process of the effect of PAMK and CTX. The above results suggest that CTX leads to the activation of the ferroptosis pathway and that PAMK has an inhibitory effect on this pathway. Therefore, we observed the microstructure of the thymus to analyze whether the activation of the ferroptosis pathway caused thymus injury. HE staining results E,F showeIn scanning electronic microscopy D, CTX caTo further clarify the involvement of ferroptosis in CTX-induced thymus injury, we detected the protein expression of GPX4 in thymus by immunofluorescence . The resp < 0.05), and the content of GSH was significantly decreased (p < 0.05) in the CTX group. In the PAMK + CTX group, the content of GSH was significantly increased (p < 0.05), and ROS levels were significantly decreased (p < 0.05), compared to the CTX group. As for the content of MDA and iron, although there was no significant difference in the PAMK + CTX group compared to the CTX group (p > 0.05), the trend was opposite to that of the CTX group. This suggests that ferroptosis may be involved in CTX-induced thymus injury in goslings by increasing levels of oxidative stress and iron in the thymus, which can be alleviated by PAMK.The occurrence of ferroptosis is closely related to the level of antioxidants and iron ions. Therefore, after confirming the occurrence of ferroptosis, we measured ROS, MDA, GSH, and Fe content . Comparep < 0.05), except for Hmox-1. The mRNA expression of the above genes was significantly lower in the PAMK + CTX group than in the CTX group (p < 0.05), except for the NRF2.CON group (p < 0.05). CTX stimulation did not cause a significant decrease in the mRNA of GPX4 but could cause a significant decrease in the protein expression of GPX4 (p < 0.05). Although the change in FPN1 mRNA expression between the CTX and PAMK + CTX groups was insignificant, the expression in the PAMK + CTX group returned to a level similar to that of the CON group. The above results suggest that PAMK can alleviate CTX-induced ferroptosis in thymocytes by blocking the activation of the ferroptosis pathway.Finally, we analyzed the specific changes in the ferroptosis pathway. We examined the relative mRNA expression of ferroptosis pathway genes and the protein expression of GPX4, a marker protein of ferroptosis . Among tFerroptosis is a new form of programmed cell death with iron-dependent properties, whose main features include lipid reactive oxygen species accumulation, iron ion accumulation, and lipid peroxidation. The metabolites of CTX after hepatic biotransformation exert a suppressive effect on the immune function of the body , bringinStudies on the cytotoxicity of CTX have mainly focused on apoptosis ,25. HoweWhen ROS accumulates in excess in cells, membrane polyunsaturated fatty acids are easily oxidized by ROS and produce MDA, for example, causing ferroptosis . GPX4 usIron homeostasis has been shown to play a critical role in the innate immunity of the organism ,37. HostGPX4 plays a very central role in inhibiting ferroptosis, but we found that CTX does not inhibit GPX4 transcription but rather GPX4 protein expression, which is consistent with previous studies . The newIn conclusion, PAMK could reduce thymus injury by alleviating CTX-induced thymocyte ferroptosis in gosling to alleviate the immunosuppression caused by CTX in the organism. Meanwhile, PAMK could co-mitigate ferroptosis in thymocytes by regulating the expression of critical genes of ferroptosis, restoring GPX4 protein expression, improving antioxidant capacity, and reducing iron overaccumulation .


Moon watching is a method of quantifying nocturnal bird migration by focusing a telescope on the moon and recording observations of flying birds silhouetted against the lunar surface. Although simple and well-established, researchers use moon watching infrequently due in part to the hours of late night observation it requires. To reduce the labor entailed in moon watching, we designed a low-cost system called LunAero that can track and record video of the moon at night. Here we present a proof-of-concept prototype that can serve as a platform for citizen scientists interested in observing nocturnal bird migration. We tested the video recording on clear nights from February 2018 to May 2019 when the moon was full or nearly full. Manual analysis of a 1.5 h sample of video revealed a total of 450 birds, which is a much higher detection rate than previous moon watching efforts have yielded. The hardware described here is part of a larger effort involving software development (currently underway) to automate recorded video analysis. We argue that LunAero can reduce the labor involved in moon watching, offer improved data quality over traditional moon watching, and provide insights into social behavior and wind-drift compensation in migrating birds. Specifications table:1Bird migration is both a fascinating and ecologically important phenomenon that influences many natural communities by transfer of biomass e.g.e.g.i.e. missed observations). Moreover, moon watching requires a considerable investment of labor and, for diurnal researchers, lost sleep. However, as noted by Liechti and Bruderer One venerable and well-tested means of observing migration is to simply focus a telescope on the moon and use it as a “backdrop” to note the passage of individual birds overhead. This observational method, known as moon watching, was first described by W. E. D. Scott, in 1881 following a fortuitous observation of flying birds during an astronomy demonstration In an effort to streamline the moon watching method, we developed a system that automates the process through use of a CCD camera, a small computer, a spotting scope, a motorized mount, and some simple computer vision techniques. The system, dubbed LunAero, performs the relatively simple task of recording high-definition video with sufficient magnification to observe and quantify nocturnal bird migration. The hardware described in this paper is part of a larger effort that involves development of software for analyzing the recorded video. This video analysis software is to be the subject of another publication. In this paper we describe a working LunAero system and present the results of testing performed during peak bird migration in the spring of 2018 along with some preliminary analyses of bird densities and flight behavior.2The LunAero system consists of a motorized, two-axis mount for a spotting scope or a telescope that has been integrated with a CCD camera via a small computer. The CCD camera is mounted to the scope such that it can record magnified images of the moon allowing the computer to both store video footage and adjust the position of the scope as needed via the motorized scope mount see . In desihttps://doi.org/10.17605/OSF.IO/N7ZEK.With this design strategy, we were able to produce LunAero from readily sourced parts and materials that are easily obtainable by research groups and citizen scientists with minimal technical experience. Detailed design files and guidelines for construction are online at Open Science Framework repository for collected data at 2.1The core mechanical portion of the automated moon tracking device is an altitude-azimuth mount that supports a wide array of telescopes. Spotting scopes in particular are a pervasive tool of amateur bird watchers, meaning that many potential users already have one. The scope mounts to a cradle with a 1/4 inch bolt, the standard for tripod mount plates 2.2A key element of the development of LunAero was finding a low-cost computer that could handle computer vision, motor control, and video recording. Prototype versions of LunAero used a Raspberry Pi 3  computer running a distribution of Raspbian Linux. However, we found that the combined demands of image processing and high-resolution video recording were too great for the Raspberry Pi. Therefore, we upgraded to an ODROID N2  to operate the hardware. The N2 is a Unix capable machine with a powerful processor and USB 3.0 ports as well as a general purpose input/output (GPIO) capable of interfacing with the TB6612FNG.LunAero used a ELP-SUSB1080P01 camera module  which is based on the Sony IMX291 sensor. The camera module was equipped with a 18 mm fixed focus “S-mount” lens to produce f/2.7 optical characteristics. This configuration produced optics that match well with the target “eye relief” or apparent exit image diameter from the ocular lens of a spotting scope.2.3The moon tracking program works by determining the position of the moon within the camera frame and occasionally activating the motors on the scope mount to bring the moon image close to the center of the field of view. To minimize shakiness in the recorded video, the scope mount remained motionless most of the time, allowing the moon to drift through the field of view. Under this scenario, the mount only needed to move at intervals of approximately 1 m.The moon tracking program was written for Python 3 NumpyPillow packages Numpy package then determines the centroid of the moon profile to index the moon’s position in the video frame. If this centroid is found too far from a reference point near the center of the frame, then the program activates the motors to correct the position of the moon image in the video frame. Motor control is effected by a software pulse-width modulator that sends signals to the TB6612FNG motor controller using the WiringPi Python package When recording initiated, the program assumes control of the motors and begins the process of maintaining the moon in the camera’s field of view. This tracking process is accomplished by extracting a low resolution frame (800 P  × 600 P) from the camera’s video stream every few seconds and converting the pixels to value arrays using the While the robotics keep the moon in view, LunAero uses OpenCV •Making it easier for researchers to observe nocturnal migration via automatic video recording.•Increasing the quality of data produced by nocturnal migration observation by allowing researchers to replay video and refine observations.•Increasing the number of observers at a given site or sites; low-cost hardware is cheaper than hiring technicians.•Retaining accessibility; hardware costs are minimized such that the technology is accessible to most research groups and enthusiasts.•Allowing future researchers to apply machine learning and computer vision technology to archived video.The hardware reported in this publication benefits researchers by:33.1The entries in e.g. a user with a large scope would need to produce large support arms.The file MoonWatcher_wood1inch.dxf is a single layer drawing to act as a guide for preparing parts from 19.05 mm thick wood, not suitable for direct upload to a cutting instrument. Instead, it should be printed at exact scale and used as a drilling/cutting guide. This design adapts to a user’s needs because the hole spacings and cut lines represent the smallest recommended parts. Therefore, the user may customize the parts, 3.2A file manifest for operation is included in Files ODROID_INSTALL.sh and odroid_moontracker.desktop are present for the convenience of the user. The install script can be executed by the user in Linux to automatically fetch and install dependencies required for the successful operation of the device. The desktop file can be copied to the user’s desktop on the ODROID computer to activate the Python script.4A more complete bill of materials including the quantity of individual nuts and bolts required for the project is present on the OSF repository in the Build Instructions subdirectory see .52 laser. While amateur ornithologists are unlikely to own personal CNC equipment, access to inexpensive cutting sources has become increasingly available. A survey from The LunAero hardware design incorporates a number of acrylic, acetal, and wooden parts, found in the 2D CAD files. Cut all acrylic and acetal parts from sheet stock using the CNC method of choice, following all instrumentation guidelines for that equipment. The plastic parts used in the unit described in this paper were fabricated on an 80 W COUsing the bolts listed on the detailed bill of materials, assemble the LunAero unit. Pictorial instructions for each step of the assembly are available at the OSF repository associated with this manuscript. Note that most of the angled connections in LunAero between flat materials use a “cruciform” bolt and nut connection, shown in When deploying LunAero, the user’s spotting scope is bolted to one of the drill holes in the cradle with a 6wget to pull the software files from the online repository. In this description, we assume that the user has downloaded the software from the repository to a file directory such that the directory tree appears similar to:To use the LunAero hardware, the appropriate software must be installed from the OSF repository. In the Linux terminal of a functional ODROID computer running Ubuntu MATE, use If your download appears different, you may experience problems with the next steps. Use the terminal to run the script at /home/%USER%/Documents/LunAero/ODROID_INSTALL.sh as a super user. This script installs all of the prerequisite files and programs required to run the LunAero tracking program. Copy the file /home/%USER%Documents/LunAero/odroid_moontracker.desktop to the desktop of your Ubuntu MATE installation. This desktop file makes it easy to activate the program without using the terminal.As explained in detail in the instructions included in the OSF repository, the user operates LunAero by acquiring a good-quality image of the moon through the camera, then activating the automatic mode. Placing the LunAero hardware on a flat surface, activate the ODROID computer. Open the moon tracking program by clicking on the desktop icon. When the program starts, go through the time-correction queries, and begin manual tracking. Use the arrow keys on the keyboard to move the scope with the motors until the screen shows the moon completely in view. Adjust the scope magnification and focus until the moon appears crisp to the computer view. Change the brightness of the output image by adjusting the digital camera exposure and ISO in the moon tracking program. The image on the screen should be bright, have sharply focused edges, and the craters of the must be equally crisp to the edge of the moon. Once you are happy with the image quality, press r to quit manual tracking mode and enter automatic mode. At this point computer vision intelligence of the robotics take over and track the bright center of the moon. Barring weather changes or extreme events, LunAero may be left alone to record the moon now. Once you done recording, transfer the video found on the hard drive of the ODROID computer to the storage medium of your choice for archival purposes.7We evaluated the hardware by field testing on nights with clear skies and a moon that was half full or more. We conducted tests in April and May of 2018 and 2019, which roughly corresponded to peak bird migration season. Tests were conducted in Norman, OK, USA, using either a 40× ATS 65 spotting scope  or 20–60×80 spotting scope (Alpen Optics). From approximately >70 h of footage collected, we selected a recording sample of 1.5 h beginning at 9:00 PM CDT on April 28, 2018 for manual annotation and analysis. Video annotation was performed by A.H. by playing the video on a desktop computer and noting the time and number of silhouettes present. During annotation, video playback was paused, reversed, and incremented frame-wise to allow for the most accurate object count and timing. This process took approximately 11.5 h to complete.In addition to this simple annotation we selected an even smaller sample of the same video (9:30 PM–10:30 PM CDT) to manually determine flight directions by measuring transit paths on a computer screen with a protractor. These measurements allowed us to emulate the moon watching protocol developed by Lowrey We counted 450 birds in the 1.5 h test recording, as well as one silhouette that was clearly an insect. The silhouettes ranged greatly in size, with the smallest silhouette scarcely larger than a few pixels see . SimilarAnalysis of flight directions revealed NW-trending net migration density, reckoned 110° north of east see . The net7.1LunAero was first conceived as a means of reducing both the labor required for moon watching and the inconvenience of nighttime data collection. However, our efforts thus far have demonstrated that LunAero surpasses the capabilities of traditional moon watching by enabling improvements in terms of the quality and types of data made available. The most obvious advancement is that LunAero remedies the problem of human error. The ability to detect birds is greatly enhanced in a frame-by-frame examination of recorded video compared to real-time bird counts. In particular, analyses performed on video footage allow for detection of birds that merely skirt the edges of the moon, passing through the lunar backdrop too quickly for most observers to discern in real time. Nisbet et al. lamented these near-edge birds as unusable data In addition to offering improvements in data quality, LunAero footage also allows us to extract new forms of data from a moon watching effort. With video analysis, the relative timing of bird observations can be determined with high accuracy, allowing for quantitative studies of flocking behavior see c. .We have demonstrated that video analysis can be used to derive precise flight directions for individual birds as well as the orientation of a bird’s body during flight . CombineA few assumptions must be made to extract information about the flying birds. The accepted approach assumes that all observed migrants are passerines between 10 g–100 g, an approach gradually codified based on original quantification of nocturnal migrants by At present, the most commonly used methods for quantifying nocturnal migration involve some form of radar. Radar provides migration data on a variety of scales ranging from the continental data provided by the NEXRAD network of weather surveillance radars, to short-range marine radars and tracking radars capable of detecting individual birds. NEXRAD generates aggregate, macro-scale data based on reflected photons that can be attributed to a group of dozens to thousands of birds within a given radar volume. Estimating numbers of birds in a radar volume requires assumptions about the radar reflectivity of bird aggregations, and these assumptions have not been fully validated •Moon watching relies on moon phase. Although we have not fully tested the limits of LunAero and the moon watching method, we think that it would only be effective even when the moon is at least >50% full.•Moon watching necessitates clear skies. Even a brief obfuscation of the moon by clouds can cause LunAero to lose its fix on the moon’s position. At present we have no means for LunAero to recover from having lost the moon, although it would be possible to implement a search pattern.•Moon watching samples a small portion of the sky. If factors such as artificial light, landscape features, or wind phenomena affect the specific routes taken by nocturnal migrants, then there is the potential for biased migration estimates depending on the particular location of moon watching equipment relative to these factors.Moon watching has several inherent limitations, even when enhanced with LunAero.Additionally, quantifying the birds recorded in LunAero video samples is time-intensive, taking at least ten times as long as real-time observation. We regard this time expenditure as a trade-off rather than a distinct limitation, attempting to quantify LunAero recordings produces more accurate results at the cost of time investment.By applying modern computing to nocturnal migration bird watching, we have advanced moon watching by not only making it more convenient, but by improving it’s accuracy and utility as a research method. Not counting the cost of a spotting scope, a LunAero system can be assembled for <$400. By keeping the cost low we ensure the practice of moon watching remains accessible and within the means of a citizen scientist with a strong interest in migration. Although moon watching has not been as widely employed as other research techniques since Lowery’s efforts in the 1950s and 1960s, we argue that our automated recording system may help this classic technique find a niche in modern migration studies.The authors would like to acknowledge funding from the University of Oklahoma’s Strategic Organization in Applied Aeroecology and donors from the LunAero crowdfunding campaign. Declarations of interest: none.The work presented in this manuscript did not involve handling or materially affecting animals. As such, it was exempted from review by the University of Oklahoma Institutional Animal Care and Use Committee.


BackgroundDuring and after the coronavirus disease 2019 (COVID-19) pandemic, there was a significant increase in the demand for aesthetic procedures nationwide. We believe that one of the factors contributing to this increase was the use of video conferencing apps. Therefore, the purpose of this study was to analyze the aesthetic concerns that were identified by video conferencing.MethodologyIn Saudi Arabia, an observational, cross-sectional study was conducted from February to April, 2022. An online survey was used to assess the aesthetic concerns raised by video conferencing. The survey addressed multiple aspects, including demographic data, information on video call utilization, the attentional focus of the participants during video calls, and new appearance concerns identified on video calls.ResultsThe survey successfully captured 432 adult participants. More than 85% of participants disliked at least one character of their appearance during video conferencing. The skin color type , time spent on video calls, and visual focus during the virtual meeting were significantly correlated with self-perception of new disliked features.ConclusionVideo conferencing showed a significant statistical correlation with identified disliked facial/body features in both genders. People with skin color type III were less likely to perceive a disliked feature through video conferencing applications. The likelihood of having a disliked feature was significantly higher among individuals who spent up to one hour per day using video calls. Additionally, those who mainly paid attention to their face or body during virtual meetings were more likely to have disliked features. Aesthetic and cosmetic procedure demand demonstrates solid global growth. This growth has been observed mainly after lockdown measures were relaxed, together with reinforcement of all necessary post-coronavirus disease 2019 (COVID-19) precautions for a safe environment .There was a sudden and dramatic increase in the use of video conferencing applications such as Zoom , Microsoft Teams , and Google Meet , due to the COVID-19 pandemic -6. The rBesides the popularity and major advantages of video conferencing apps, it has a detrimental effect on how individuals perceive their appearance . Video cDue to the closure of many cosmetic/beauty services and limited accessibility to aesthetic procedures during the lockdown, people with high dysmorphic concerns were reported to have worsened appearance dissatisfaction ,14. ConsVideo conferencing will likely be used long after the pandemic because studies indicate that by 2024, only 25% of corporate meetings will be held in person . ConsequA cross-sectional study was conducted in Saudi Arabia between February and April 2022. The study received ethical approval from the Medical Research Ethics Committee at Shaqra University, Saudi Arabia (CMD/DWD/SU/2022/03/066). Participants were eligible to participate if they were 18 years or older and currently residing in Saudi Arabia. The sample size was calculated using the SurveyMonkey  calculator with a confidence interval of 95% and a margin of error of 5%, which yielded a sample of 385 subjects. Data collection was performed using an online questionnaire to collect the data and was created using Google Forms. The questionnaire was distributed on social media platforms Twitter , Telegram , and WhatsApp , with an explanatory cover for participation and written consent.The questionnaire comprised three main sections. Section A was aimed at gathering data on the demographics of the participants, which included . Section B aimed to solicit information on video call utilization . Section C was designed to assess the participants' attentional focus and their new appearance concerns identified on video calls. Participants were asked what they paid most visual attention to during the call, whether or not they had noticed a new aspect of their appearance that they disliked due to video calls, and to specify what these new areas of appearance concern were via checkboxes. Finally, the participants were asked to indicate whether they had engaged in any behaviors related to video manipulation to improve their appearance or to feel more comfortable on video calls.Statistical analysisWe used SPSS Statistics for Windows, version 25  for data analysis. We started the data analysis by running frequencies for the demographic information and characteristics of the study sample, followed by running frequencies for the most commonly detected features by the participants as disliked traits to gauge the extent of the problem. Moreover, the adaptive techniques taken by the participants were also analyzed and presented in frequencies. A chi-square test was used to identify the relationship between different variables and their effect on the self-perception of new disliked traits after the increased demands for video conferences. Finally, a logistic regression model was created to measure the direction and the exact effect of statistically significant variables on self-perception.The total number of participants in this study was 432, distributed across the different regions of Saudi Arabia. Most of the sample was collected from the central region (50.2%). This study was composed of an almost equal distribution of females and males. Participants' age group mainly fell between 18 and 34 years, representing a total of 74.4%. More than 50% of the participants were employed; more details are given in Table This study aimed to analyze whether or not the increased demand for video calls adversely affected participants' perception of their appearance. We found that skin aspects, such as color and pimples, were the most bothering aspects reported by the participants (39.6%). At the same time, nose features, such as shape and size, were the second most disliked aspect identified during the video calls Table .Participants had to use some adaptive techniques to improve their appearance or to feel more comfortable during video calls, such as camera position and angle manipulation (45.6%), light adjustment (41.4%), and using in-app filters 39.8%) , time spent on video calls, and visual focus during the call were significantly associated with the self-perception of new disliked characteristics (p < 0.01). It is worth mentioning that 52% of the people who reported no new disliked self-aspect had not used video calls on a daily basis.A logistic regression model was performed to determine the effect of positively correlated factors with self-perceived disliked traits Table . People The demand for aesthetic procedures has increased more than usual since the COVID-19 crisis. Simultaneously, there was a dramatic increase in using video conferencing applications, such as Zoom and Microsoft Teams. In Saudi Arabia, government and private services, such as education, work, and telemedicine, were switched from in-person interaction to virtual platforms using the applications mentioned earlier. However, the association between the increased demand for aesthetic procedures and virtual conferencing has not been investigated in Saudi Arabia. In this study, we aimed to analyze the aesthetic concerns identified by using virtual meeting applications. We believe that the aesthetic concerns identified through video conferencing are one of the factors beyond the surge of aesthetic procedures. Several studies have proven that staring at self-image is linked with appearance dissatisfaction -20. ThusIn our study, we found that 85.4% of the participants identified at least one disliked feature through video conferencing, and the vast majority turned the camera on during the meetings. Thus, they were exposed to self-image staring and identifying new disliked facial/body aspects. Furthermore, we found that visual focus during virtual meetings was a significant factor associated with identifying a disliked feature. The participants who were focusing only on their faces were 15% more likely to have aesthetic concerns relative to those who were focusing on others or screen activities.In the current study, most of the disliked features identified by video conferencing were associated with facial appearances, such as skin blemishes  and the nose . That is not surprising as the above-shoulder area is the most visible area during video conferencing calls. A recent study found that public interest in aesthetic procedures for the above-shoulder area has significantly increased after COVID-19 in the United States . This fiOur study revealed that participants’ age was not correlated with a new disliked feature due to using virtual meeting applications. This suggests that the identified disliked features were not related to aging changes and were probably due to other cosmetic reasons.It was recognized that females tended to be more interested in cosmetic procedures and to have aesthetic concerns about their appearance based on studies done in Saudi Arabia before the COVID-19 pandemic -24. HoweOur data revealed that the average daily time spent on virtual meetings was a significant factor associated with identifying a disliked facial/body feature. We found that people who spent less than 30 minutes a day were 3% more likely to have a disliked facial/body feature relative to participants who were not using virtual applications on a daily basis. The risk increased to 6% in those who spent up to one hour daily. The risk was more with people who spent more than two hours, but not significant relative to non-daily users. However, there is a variation between studies regarding the duration of time using virtual applications and appearance dissatisfaction ,9,15,27.Skin color type  was the third correlated factor with self-perceived disliked features. Interestingly, participants with skin color type III were 32% less likely to perceive a disliked feature compared with type I. In contrast, participants with skin color type V were more likely to have a disliked feature, but it was not statistically significant. The reason behind that was not understood, which may need further investigations in terms of aesthetic concerns and dysmorphic disorders.This study also had some limitations. Body dysmorphia was not addressed through our survey; hence, those who were focusing on themselves during virtual meetings could be comorbid with body dysmorphia. Further investigation on the effect of video conferencing on those already diagnosed with body dysmorphia is necessary. Another limitation we can address here is the unreliable results associated with the Fitzpatrick skin type and Saudi population due to the self-assessment nature used in the online questionnaire.During the COVID-19 pandemic, video conferencing usage significantly increased; the demand for aesthetic procedures was found to be increased as well. Our study aimed to analyze the aesthetic concerns identified through video conferencing applications. We found that video conferencing correlated significantly with identified disliked traits, as 85.4% of participants identified at least one disliked facial/body feature regardless of age and gender. Moreover, skin color type , time spent on video conferencing, and visual focus during the call were significantly associated with self-perception of a new disliked feature. This study encourages the scientific society to conduct more research on this topic, primarily focusing on possible comorbid body dysmorphia.


In humans, these enzymes impact many physiological and pathological processes including respiration, pH and CO2 homeostasis, electrolyte secretion, gluconeogenesis, ureagenesis, lipogenesis, bone resorption, and tumorigenicity. For this reason, several human carbonic anhydrases have become therapeutic targets for the treatment of many disorders. In recent years, a huge number of carbonic anhydrase inhibitors have been developed for therapeutics aims, such as diuretic, antiglaucoma, antiobesity, and anticonvulsant agents, and for the diagnosis and treatment of cancer diseases. The authors report a combined crystallographic and computational study on a promising class of carbonic anhydrase inhibitors to clarify their mechanism of action and to obtain useful information for the drug design of new effective and selective molecules.Carbonic anhydrases are a family of enzymes that catalyze an essential physiological reaction for living organisms: the reversible conversion of COAliphatic sulfonamides are an interesting class of carbonic anhydrase inhibitors (CAIs) proven to be effective for several carbonic anhydrase (CA) isoforms involved in pathologic states. Here we report the crystallographic structures of hCA II in complex with two aliphatic sulfonamides incorporating coumarin rings, which showed a good inhibition and selectivity for this isoform. Although these two molecules have a very similar chemical structure, differing only in the substitution of the two aliphatic hydrogen atoms with two fluorine atoms, they adopt a significantly different binding mode within the enzyme active site. Theoretical binding free energy calculations, performed to rationalize these data, showed that a delicate balance of electrostatic and steric effects modulate the protein-ligand interactions. Data presented here can be fruitfully used for the rational design of novel and effective isozyme-specific inhibitor molecules. D (θ-CAs)  all cont (θ-CAs) .All human CAs (hCAs) belong to the α-family, which is the most populous, with 15 isoforms differing for molecular features, kinetic properties, oligomeric arrangement, and cellular localization ,13. In d2 to bicarbonate ion [A huge number of structural studies have been carried out on α-CAs, showing that, in agreement with their high sequence homology, all of these enzymes have a very similar three-dimensional structure, regardless of their oligomeric state or cellular localization. The main structural features are represented by a central twisted β-sheet enclosed by helical regions and additional β-strands ,15. The 2 molecule, leading to the formation of HCO3− which is subsequently displaced from the catalytic site by a water molecule (Equation (1)). Subsequently, in the second and rate limiting step, the zinc-bound hydroxide ion is regenerated by the transfer of a proton from the zinc-bound water molecule to the bulk solvent (B) (Equation (2)) [The hydration reaction proceeds through a two-step mechanism. The first step consists of a nucleophilic attack by the zinc-bound hydroxide ion on the COEquation ). Subseqion (2)) .EZn2+-O2 substrate and its correct orientation in the active site in order to undergo the nucleophilic attack by the zinc-bound hydroxide ion. On the other end, the hydrophilic region enables the assembly of a well-ordered hydrogen-bonded solvent network that assists the transfer of the proton from the zinc-bound water molecule to the bulk solvent. The simultaneous presence of these two regions inside the active site allows the rapid catalytic cycling of CO2 to bicarbonate [The availability of the crystallographic structures of hCA isoforms allowed the identification of two very different environments within the enzyme active site cavity: one region delimited by hydrophobic amino acids and one mainly constituted by hydrophilic residues . Severalarbonate .2 homeostasis, respiration, transport of CO2 and HCO3−, biosynthetic reactions such as lipogenesis, gluconeogenesis and ureagenesis, bone resorption, electrolyte secretion, calcification, and many others [hCAs are extensively distributed in different tissues and organs, where they are involved in various physiological processes including pH and COy others . Consequy others ,15. For To date, the most investigated CA inhibitors (CAIs) are aromatic/heterocyclic sulfonamide derivatives A, which 2 group, generally higher with respect to that of the aromatic/heterocyclic sulfonamides [1 and 2  glycerol before being flash-cooled in liquid nitrogen. Diffracted intensities were processed using the HKL2000 program [1 space group (PDB code 4XE1) [o| − |Fc| maps after checking their hydrogen-bonding geometry. Statistics for refinement are reported in The complexes were crystallized at 20 °C using a procedure previously described for other hCA II/inhibitor complexes . In deta program . Data prde 4XE1) . For botde 4XE1)  and CNS de 4XE1) ,31, respde 4XE1) . Water m2 crystallographic complex and on the three model adducts hCA II/ZBG2, hCA II/ZBG,2* and hCA II/ring2*. The model hCA II/ZBG2 is identical to the corresponding crystallographic complex hCA II/2 with the coumarin ring substituted with a hydrogen atom, whereas hCA II/ZBG2* is identical to hCA II/1, apart from having the coumarin ring substituted with a hydrogen atom and the other two hydrogen atoms substituted by fluorine atoms. The model hCA II/ring2* is identical to the crystallographic structure hCA II/2 apart from having the coumarin ring rotated of 180° thus resembling the orientation of the tail in the hydrogenated derivative 1. All the model adducts were built with Insight II software .Theoretical calculations were performed on the hCA II/Concerning the ligands, their partial atomic charges were determined using the restrained electrostatic potential (RESP) protocol implemented in the PyRED server  through 2+ ion, the Van der Waals parameters  = 0.00330286) from the work of Li and Merz [per-residue decomposition of the binding free energy was performed.The molecular mechanics/generalised Born surface area (MM/GBSA) ,36 methoand Merz  were useand Merz ,42, the bind represents the binding free energy and Gcomplex, Gprotein, and Gligand are the free energies of complex, protein, and ligand, respectively. In particular:bind is the binding free energy in solution; ΔEMM is the molecular mechanics energy including van der Waals (ΔEvdW) and electrostatic (ΔEelec) contributions; and ΔGsol is the solvation energy, and is the sum of electrostatic (ΔGGB) and nonpolar (ΔGSA) interactions. TΔS represents the entropic change due to ligand binding. Our calculations do not include this entropic term, since it is reasonable to exclude it when comparing similar ligands [GB is the electrostatic solvation energy and is computed by the Generalized Born method [According to MM/GBSA method, the binding free energy was estimated as follows: ligands ,43, in a ligands ,44. ΔGGBn method , whereasn method .1 and 2 were determined to resolutions of 1.67 Å and 1.56 Å, respectively.The crystal structures of hCA II in complex with compounds o-Fc| and |2Fo-Fc| electron density maps   see , their ac cavity .1 was an hCA II inhibitor three times more potent than compound 2, suggesting that its binding conformation was energetically favoured with respect to that adopted by the fluorine derivative. Thus, we wondered why the latter did not adopt the same binding mode shown by compound 1. To answer this question, binding free energy calculations were performed, using the MM/GBSA theoretical method [per-residue basis, to identify key protein residues responsible for the inhibitor binding mode. With the aim of evaluating the energetic effects separately due to the different ZBG positions and coumarin ring rotation observed for the two inhibitors, the calculations were carried out on the hCA II/2 crystallographic adduct and on three other model adducts, hereafter indicated as hCA II/ZBG2, hCA II/ZBG,2* and hCA II/ring2*. ZBG2 and ZBG2*, represent simplified models of the two inhibitors (2 and 1) including only the ZBGs. In particular, the model hCA II/ZBG2 corresponds to the crystallographic structure hCA II/2 with the inhibitor coumarin ring substituted with a hydrogen atom, whereas hCA II/ZBG2* is identical to the hCA II/1 crystal structure apart from having the ring substituted by a hydrogen atom and the other two hydrogen atoms substituted by fluorine atoms, thus representing a hypothetical model in which the ZBG of the fluorinated derivative 2 would adopt the same binding position observed for the corresponding hydrogenated derivative. Finally, hCA II/ring2* is the hCA II/2 crystallographic structure with the coumarin ring of 2 rotated 180°, thus corresponding to the orientation of the tail in the hydrogenated derivative 1.Previously reported inhibition studies  showed tl method ,36. ThisZBG2 and hCA II/ZBG2*; 2+ ion [Firstly, we compared results obtained for hCA II/; 2+ ion ,49.ZBG/2* with respect to hCA IIZBG/2 . Indeed, in hCA II/ZBG,2* the substitution of the short CH bonds (1.1 Å) with the longer CF ones (1.3 Å), leads to a reduction in the distances between one of the inhibitor fluorine atoms and the Thr200 side chain atoms (vdW) term and for the increase in the electrostatic (ΔEelec) term, that in hCA IIZBG/2 was already disadvantageous  are involved in stabilizing interactions with residues Ile91, Gln92, and Val121; however, in the case of ring2*, these interactions are weaker with respect to 2. Moreover, ring2* is affected by strong destabilizing interactions with Phe131 and Pro202  mainly due to van der Waals contributions (see Subsequently, calculations were carried out on the crystallographic adduct hCA II/ions see . ring2*, the coumarin ring and its methyl substituent are too close to the side chains of Phe131 and Pro202, respectively, leading to steric repulsion. These results indicate that the fluorinated compound 2 would experience significant destabilizing interactions with Pro202 and Phe131 if its coumarin ring would adopt the same orientation as the corresponding hydrogenated derivative 1.Indeed, as shown in 2-SO2NH2, the replacement of two aliphatic hydrogen atoms with two bulky and electronegative fluorine atoms can have important effects on the binding conformation not only locally at the bottom of the active site, but even remotely at the entrance of the catalytic cavity, thanks to a delicate balance of electrostatic and steric effects involving enzyme residues.Overall, our energetic calculations show that in aliphatic sulfonamides with general formula R-CH2-SO2NH2 type, the introduction of substituents on the aliphatic carbon atom can be used to modulate the affinity of the inhibitor against a specific CA isoform. In this respect, the present study represents a step forward in the rational design of novel isozyme-specific inhibitor molecules with improved features.In conclusion, the combined crystallographic and computational study reported here describes in detail the structural features and the energetic factors involved in the interaction of hCA II with a promising class of aliphatic sulfonamides incorporating coumarin rings. In agreement with previous studies ,42,50, i


Studies have not provided clear enough evidence on the direct association between cigarette smoking and poverty. This study aims to assess the association of householder smoking with near-poverty households, and the potential mediating effect of NCDs.2-test was performed, and multivariable logistic regression and extended probit regression models were fitted to examine the association between householder smoking and near-poverty households. Moreover, generalized structural equation modeling was used to explore the mediating effect of NCDs.A cross-sectional survey was conducted from November 2019 to October 2020 in relatively underdeveloped regions in China. In total, 2,409 households were investigated in areas under the jurisdiction of 24 primary health care (PHC) institutions of eight provinces. Pearson's χAfter adjusting for all other potential confounding factors, compared with households headed by never-smokers, households headed by smokers exhibited significantly elevated risks of being near poverty, with an odds ratio of 2.01 (95% CI: 0.48–0.91). We also found that living in rural areas and having a low education level both had a negative effect on being near poverty. Additionally, NCDs had a significantly positive mediating effect, with a 31.57% effect of householder smoking on near-poverty status mediated by NCDs; the indirect effect was estimated to be 0.17 (95% CI: 0.04–0.31).Householder smoking significantly elevated the risk of the household being near poverty, and suffering NCDs had a positive mediating effect. Cigarette smoking is a major public health problem and remains the leading preventable cause of death and disability in China and other countries, killing more than 8 million people a year around the world . In 2020Additionally, even though the global percentage of people living in extreme poverty declined from 36% in 1990 to 10% in 2015, more than 700 million people now live in extreme poverty , 9. MeanAt the macro level, multiple studies have demonstrated that tobacco use imposes an economic burden through a reduction in productivity and an increase in the cost of medical treatment . For exaAt the micro level, there have been few studies about cigarette smoking and poverty. Liu et al.  estimateWe conducted a cross-sectional questionnaire-based survey in relatively underdeveloped regions in China from November 2019 to October 2020, and the participants were enrolled using a stratified multistage sampling method. According to per capita GDP levels, we first chose 12 cities/counties that had relatively lower per capita GDP in eight province-level regions in China . Then, in each city/county, two primary health care (PHC) institutions were randomly selected as our investigation units, for a total of 24 PHC institutions.Next, with the coordination of local health professionals in PHC institutions, approximately 100 households were selected in areas under the jurisdiction of each PHC institution. The participants were not sampled randomly but were directly selected based on the following: (1) living in local communities for at least 6 months; (2) willing to participate in this study; (3) the householder was aged over 40; (4) near-poverty households and nonpoor households were included; and (5) smokers and never-smokers were included.The selected householders had a face-to-face interview with the local health professionals of the PHC institutions, and prior to the interview, investigators received professional training to ensure data collection quality. A validated interviewer-administered questionnaire was used to obtain information about the household's basic situation , demographic characteristics of the householder , and the householder's smoking status, whether the householder suffered from NCDs or not and so on. All measures were self-reported in the study.In 2019, the Chinese government defined the poverty as yearly per capita income <3,747 Yuan (or US $1.49 per day). In our study, we further defined near-poverty as 200% of the poverty level definition  and categorized household poverty status as near-poverty  or nonpoor .Smoking status was categorized as smoker (including current smoker and former smoker) or never-smoker. Participants were asked: “Have you smoked in the last month?” When the respondent answered “yes”, the individual was categorized as a “current smoker”; if the respondent answered “no”, he or she was then asked, “have you ever smoked?” If the respondent answered “yes”, the individual was considered to be a “former smoker”; if the answers to both questions were “no”, the respondent was considered a never-smoker.Noncommunicable disease status were determined by the following question: “Have you ever been diagnosed with noncommunicable diseases, such as hypertension, diabetes, COPD, CHD, strokes, cancer and so on?” Respondents who replied “yes” were categorized as having NCDs.We controlled for an array of demographic and socioeconomic statuses that have been previously shown to be associated with cigarette smoking and poverty. The basic household situation included the household's residential location  and household size (number of household members). Demographic characteristics of the householder included the following: sex ; age group ; educational level , marital status  and alcohol use (yes or no). We also asked, “Does raising cigarette prices influence tobacco use?” (yes or no). All covariates were categorical variables.2-test was performed to assess differences in the characteristics between near-poverty and nonpoor households for the categorical variables. Both univariate and multivariate logistic regression models were used to determine the association between householder smoking and near-poverty households. The covariates were adjusted in the multivariable logistic regression model. Moreover, to accommodate endogenous covariates, we fitted an extended probit regression model  variables were described using frequencies and percentages. Pearson's χon model  using thP < 0.001). NCDs were reported to be more prevalent among near-poverty householders (51.16%) than nonpoor householders (30.32%). Furthermore, near-poverty householders tended to be older, to be single or divorced/widowed, and to have a lower education level. In terms of householder sex, there were no considerable differences between the two categories of households .The study included 1,595 households from rural areas and 814 households from urban areas. Of the 2,409 households, 430 (17.85%) households were near-poverty, and 1,979 (82.15%) households were nonpoor. A total of 1,183 (49.11%) householders were never-smokers, while 1,226 (50.89%) householders were smokers. The ages of the householders ranged from 40 to 80 years, with an average age of 55.52 ± 10.42 years. P < 0.05). As shown in Models 1–2, compared to households headed by never-smokers, households headed by smokers had an approximately 50% increase in the probability of household being near poverty, with odds ratios of 1.57 (95% CI: 1.27–1.94) and 1.49 (95% CI: 1.14–1.95), respectively. In Model 3, we found that the correlation between the errors of our two equations was significantly negative, so the unobservable factors that increased the householder smoking rates decreased the probability of households being near poverty. The results of Model 3 revealed that households headed by smokers had a two times higher risk of being near poverty than households headed by never-smokers, with an odds ratio of 2.01 (95% CI: 0.48–0.91).Moreover, households in rural areas had higher odds of being near poverty than households in urban areas . Additionally, we also found that the higher the education level of the householder was, the lower the predictive probability of the household being near poverty in both rural and urban areas. However, regarding different education levels, households headed by smokers all had higher probabilities of being near poverty than households headed by never-smokers .In this study, we observed a strong association between householder smoking and household being near-poverty status that persisted after adjusting for basic household situation and sociodemographic factors. In line with previous studies , 21, ourMoreover, our study revealed a higher impact of smoking on poverty than in a previous study . PossiblThe purchasing of cigarettes imposes a financial burden on low-income smokers and their families, but tobacco-related diseases, such as NCDs, exact an even greater cost and plunge those already on the margins into poverty. This study revealed that NCDs had a significantly positive mediating effect between householder smoking and household being near-poverty status, which offered evidence to consider the underlying mechanisms of the observed association. Potential explanations include the toxic effects of smoking on the human body, including but not limited to endothelial dysfunction, inflammation, and cancer, leading to a higher chance of developing NCDs , 24. FurOur study indicated that cigarette smoking has a strong association with near-poverty status, so from a policy perspective, controlling tobacco use and reducing smoking prevalence are not only important public health issues but are also closely related to poverty alleviation. As the most populated developing country in the world, China has made remarkable progress in poverty alleviation; however, problems of “sickness poor” and “poverty due to illness” remain the biggest barriers to consolidating achievements in poverty alleviation. Therefore, it is recommended to integrate tobacco control strategies with national poverty alleviation policies, which will be helpful in overcoming economic and political obstacles in the implementation of existing tobacco control measures. Additionally, we also found that NCDs mediated the association between householder smoking and near-poverty status, so it is recommended to strengthen health education and expand the coverage of medical insurance, which will be useful to lighten near-poverty households' financial burden and solve near-poverty households' inclination toward poverty because of illness. Furthermore, worldwide evidence has shown that raising tobacco taxes had the single greatest impact on the reduction of tobacco consumption , 28, andWe also noted that a low educational level of the householder was significantly associated with a higher probability of being near poverty. Previous studies have shown that less-educated individuals and individuals with lower socioeconomic status were more likely to be smokers , 32. In The strengths of our study include the nationally representative survey. A broad range of data from relatively underdeveloped regions in China was collected, supplementing the empirical research conclusions on the relationship between householder smoking and household being near-poverty status. Moreover, in the study of the effects of householder smoking on near-poverty status, data endogeneity was unavoidable, and an extended probit regression model with the instrumental variable was used to overcome endogeneity problems. In addition, we used generalized structural equation modeling to evaluate the mediating effect of NCDs, providing evidence to consider the underlying mechanisms of the association between householder smoking and near-poverty households. However, there are also several limitations in this study. First, this study is a retrospective self-reported survey, and recall bias may be inevitable; however, prior studies have found that self-reported smoking status and health conditions had reasonable validity , 38. SecIn conclusion, householder smoking significantly elevated the risk of households being near poverty, and suffering NCDs had a positive mediating effect between householder smoking and near-poverty status. We also found that a lower educational level and living in rural areas were significantly associated with a higher probability of being near poverty.At the macro level, tobacco use imposes an economic burden through a reduction in productivity and an increase in the cost of medical treatment.At the micro level, few studies estimate the impact of cigarette smoking on poverty by reducing the income of Chinese urban residents and excessive medical spending.At the micro level, householder smoking significantly elevated the risk of households being near poverty.Suffering from NCDs has a positive mediating effect on the relationship between householder smoking and households being near poverty.yanghuimin85@126.com.The datasets presented in this article are not readily available because data involves personal privacy issues. Requests to access the datasets should be directed to HY, This study was approved by the Ethics Committee of the Capital Institute of Pediatrics, Beijing (ID: SHERLL2020017). The patients/participants provided their written informed consent to participate in this study.CJ, BC, and HY conceptualized the study. The questionnaire was developed by CJ, BC, and AG and refined with input from HY. Data collection in the field was coordinated by BC and AG and supervised by HY and JS. HY, JS, and XC contributed substantially to data analyses and interpretation of the results under the guidance of CJ. HY drafted the manuscript. CJ and BC revised the manuscript critically for intellectual content. All authors approved the final version of the manuscript.This work was supported by the Capital Institute of Pediatrics's Research Foundation (QN-2020-21) and the Capital's Funds for Health Improvement and Research (CFH 2020-3-1132).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The aim of this study was to compare the angiographic appearance of symptomatic and asymptomatic carotid near-occlusion. We have found no such previous study. The study hypothesis was that among symptomatic patients with ≥ 50% carotid stenosis, near-occlusion is more common and near-occlusions are more severe than among asymptomatic persons with ≥ 50% carotid stenosis. We reassessed consecutive CTAs from 4042 persons, 645 had ≥ 50% carotid stenosis, and 385 (60%) symptomatic. Near-occlusion was similarly common in symptomatic  and asymptomatic  cases. Among near-occlusions, the angiographic appearance was very similar between symptomatic and asymptomatic cases: mean stenosis lumen diameter (0.7 mm), distal ICA diameter (2.1 mm), and ECA ratio (0.79) were the same in both groups. Mean ICA ratio (0.46 and 0.48) and share of full collapse was very similar (45% and 42%). These findings add to the pathophysiological understanding of carotid near-occlusion. Carotid near-occlusion is a severe carotid stenosis that causes a size reduction (collapse) of the ICA beyond the stenosis . Near-ocWe reassessed consecutive CTA exams from 4042 persons aged ≥ 18 years from 2010 to 2014. We included all persons with extracranial ≥ 50% carotid stenosis. We reviewed the medical records and defined symptomatic as cases with an ischemic cerebrovascular event fitting with the distribution of a stenosed artery within 6 months of the exam. Three observers  reviewed the medical records blinded to degree of stenosis, but side of stenosis was known as this was needed for data synthesis. All cases with unclear symptomatic status were assessed by a neurologist with extensive carotid stenosis experience (EJ). Diagnosis of near-occlusion was done as described elsewhere by two observers (EJ (2 years experience), and AJF (> 40 years experience)), blinded to each other and to clinical data and with inter-rater observer kappa of 0.80 . Disagret-test and χ2-test in bivariate analyses. In multivariable analyses (linear and binary logistic regression), we used all parameters with p ≤ 0.1 in the baseline assessment as co-variates. p < 0.05 was considered statistically significant. A Bonferroni-corrected p-value threshold was calculated as 0.05/m for the two analyses with multiple testing , nor between near-occlusion and conventional ≥ 50% stenosis . Among near-occlusions, there were no differences in any parameter of how severe the near-occlusion was between symptomatic and asymptomatic cases in bivariate and multivariable analyses  were symptomatic. There were several baseline differences between the symptom groups, albeit some might have been false positive due to multiple testing Table . CompariThe main finding of this study was that there was no difference in angiographic appearance of near-occlusions between symptomatic patients and asymptomatic persons.The mechanism of the recurrent strokes in near-occlusion with full collapse is unknown. Cerebral hypoperfusion has been proposed –7. As ICStrengths of this study were the large sample size and rigorous classification of symptom status and degree of stenosis. Weaknesses include lack of test of cerebrovascular reserve and possible selection bias among asymptomatic stenoses, as many were detected by cerebrovascular disease and more symptomatic patients came by referral than asymptomatic persons.In summary, there is no difference in angiographic appearance between symptomatic and asymptomatic carotid near-occlusions. These findings add to the pathophysiological understanding of carotid near-occlusion.


Streaming services provide people with a seemingly infinite set of entertainment choices. This large set of options makes the decision to view alternative content or stop consuming content altogether compelling. Yet, nearly all experimental studies of the attributes of video content and their ability to influence behavior require that participants view stimuli in their entirety. The present study measured neurophysiologic responses while participants viewed videos with the option to stop viewing without penalty in order to identify signals that capture the neural value of content. A post-video behavioral choice was included to reduce the likelihood that measured neurophysiologic responses were noise rather than signal. We found that a measure derived from neurophysiologic Immersion predicted how long participants would watch a video. Further, the time spent watching a video increased the likelihood that it influenced behavior. The analysis indicates that the neurologic value one receives helps explain why people continue to watch videos and why they are influenced by them. Audiences want to be immersed in content, from movies to TV, to music , arousal (anterior cingulate), and executive function . Define n as the neural response to the stimulus n ∈ ℝ+, let scalar d > 0 be a neural threshold parameter, and let t ≥ 0 denote time. Then, the sigmoid function σ:ℝ→ maps the neural response and time for a given threshold d into a probability,In order to generate a testable hypothesis, we propose a simple mathematical model relating neurophysiologic responses to post-stimulus decisions. The model is a static version of the classic Hodgkin-Huxley model of the propagation of action potentials between neurons  and t. A key prediction of the model is that the likelihood of deciding to act remains low when the neural response is small, but the cumulative effect of the neural response increases with time. The behavioral response is driven by the hysteresis that occurs at threshold d in which the probability rapidly changes from low to high. The model predicts that both time and neural responses increase the probability of taking an observable action and that the probability of an action rises after threshold d is passed. We will call threshold d “Acceleration” to make the analysis easier to read. The model guides the empirical approach that operationalizes Acceleration, described below. It also indicates that time will mediate the impact of neurophysiologic responses on behavior, further circumscribing the empirical analysis. The probability of taking an action increases with neural activation in response to the stimulus; that is, M = 28.01, SD = 12.64). This study was approved by the Institutional Review Board of Claremont Graduate University (#3444) and all participants gave written informed consent prior to inclusion. The data were anonymized by assigning an alphanumeric code to each person. After consent, participants were seated at partitioned computer stations with headphones and were fitted with PPG (photoplethysmography) sensors . They then completed a short survey on demographics and mood using Qualtrics software .Sixty-two participants  were recruited from the Claremont Colleges and surrounding community using our standing subject pool . Neurologic Immersion combines signals associated with attention and emotional resonance to a stimulus that was identified in studies of neurochemical and electrical signals that predict social behaviors, including charitable donations  was used to present the stimuli and to synchronize choice termination with physiologic data. The video order was counterbalanced.n in the mathematical model. The threshold d (Acceleration) in the theoretical model was operationalized using the cumulation of neurologic Immersion above a threshold value, similar to previous research . Parametric relationships were examined using correlations and logistic regressions. A logistic regression with bootstrapped standard errors was estimated to establish the predictive accuracy of the mathematical model. Since the mathematical model predicts that time and neurophysiology both influence the donation decision, a mediation model was estimated to examine this relationship. Finally, sensitivity analyses were conducted by adding control variables age, sex, income, and positive and negative affect to determine if these factors influenced the time watching the video and/or the donation decision.The data are an unbalanced panel due to differing video stop times. The analysis begins with tests of mean differences between donors and nondonors using Student’s N = 550). The average donation varied substantially across videos, from $0 to $0.46. On average, participants donated to 1.44 of the 11 charities shown in the videos (SD = 1.90). Whether a video elicited a donation also varied substantially, from videos with zero donations to 15 participants who donated money for the most effective video. There were no differences in the IRI subscales of personal distress (PD) or empathic concern (EC) between those who donated money compared to those who did not donate . Participants reported they felt closer to issues to which videos they donated . IOS was significantly correlated with donations .The average donation across all 11 videos was $0.21 , and age . Biological sex and education did not affect donations .The total amount donated across all videos increased with participants’ incomes  as did the proportion of participants who watched videos in their entirety . Indeed, the data revealed positive correlations between time watching a video and whether a donation was made .The free choice of time spent watching each video showed substantial variation, from 9 s to 444 s . However, average Acceleration was higher when donations were made  with the time watching videos positively related to Acceleration  while Immersion was not .Average neurologic immersion while watching videos was unrelated to the donation decision . The estimation produced a log-odds ratio of 1.0017. That is, a one-unit increase in Acceleration while watching a video was associated with a 0.17% increase in the likelihood of a donation. The Acceleration continued to be positive and significant  and has a higher log-odds ratio (1.0025) when including as controls an indicator for video finishing (VF), age, income, sex, and IOS .We estimated a logistic regression to assess the individual impacts of Acceleration and time on donation decisions. Acceleration was significantly associated with donations  with a similar log-odds ratio (1.004). The time watching a video continues to be positive and significant  when controls are included , with an increased log-odds ratio (1.005).Time also had a positive effect on the donation decision in a logit estimation . This reveals another factor that affected how long participants watched a video. The Acceleration-closeness relationship may reflect “top-down” control in which additional neural resources are devoted to processing relevant information. For example, a functional brain imaging study showing photographs of objects documented greater neural activation when smokers viewed a picture of a cigarette pack compared to nonsmokers’ responses . The median age of participants was 25 while only 7% of sample was over 60 years old so this effect may be an artifact of the study population. Yet, there was no relationship between Acceleration and age  indicating that our findings were not driven by the behavior of the oldest participants.Curiously, the average time watching videos increased with participant age  and all participants gave written informed consent prior to inclusion.PZ: conceptualization, methodology, and software. RN: data collection. L-HL and RN: formal analysis. L-HL and PZ: writing—original draft, writing—review and editing. All authors contributed to the article and approved the submitted version.