Bone disorders affect millions of people worldwide and treatments currently available often produce undesirable secondary effects or have limited efficacy. It is therefore of the utmost interest for patients to develop more efficient drugs with reduced off-target activities. In the long process of drug development, screening and preclinical validation have recently gained momentum with the increased use of zebrafish as a model organism to study pathological processes related to human bone disorders, and the development of zebrafish high-throughput screening assays to identify bone anabolic compounds. In this review, we provided a comprehensive overview of the literature on zebrafish bone-related assays and evaluated their performance towards an integration into screening pipelines for the discovery of mineralogenic/osteogenic compounds. Tools available to standardize fish housing and feeding procedures, synchronize embryo production, and automatize specimen sorting and image acquisition/analysis toward faster and more accurate screening outputs were also presented. Human disorders characterized by a progressive loss of bone mineral density  or a gradual deterioration of bone architecture and deformation of bone shape  affect a large number of people worldwide, resulting in reduced patient well-being and imposing a large financial burden upon society ,2,3. SevIn the long process of drug development, screening and preclinical validation are commonly performed using rodents (mostly mouse) as they are evolutionarily close to humans . HoweverThe quest for systems that could bridge the gap between simple high-throughput cell-based assays and complex low-throughput whole-animal assays identified small aquatic vertebrates as promising tools for the screening of compounds , particumodus operandi—including turnaround time, system throughput, operating skills and requirements in compounds—and the cellular complexity greatly varies among the different systems available but a major bottleneck common to all systems is probably the extensive time required to image the bone structures and assess their morphometry  and density  through image analysis. Key processes have recently been optimized and automated, procedures in animal production have been progressively standardized, and sophisticated tools have been developed to increase the throughput and the accuracy of the zebrafish screening systems. This review presents the zebrafish systems that can be used to screen for mineralogenic/osteogenic compounds and highlights the tools available to increase their throughput for faster and more accurate preclinical studies.The first approaches to screen for osteoactive compounds were largely empirical but several in vitro , ex vivo , and in vivo  systems have now been developed and recently optimized (see description below). The assessment of the mineralogenic/osteogenic potential of selected compounds is mostly based on the morphometric analysis of bone structures stained with bone-specific dyes  or marked with reporter proteins  expressed by transgenic zebrafish lines (see description below). The Many cell lines of fish origin are available (856 in the Cellosaurus version 42 of June 2022 ) to studThe use of zebrafish embryos and larvae for compound screening and drug discovery brings several advantages in relation to throughput and ethics. Zebrafish embryo and larvae are small (approximately 0.7 mm for a fertilized egg and 4.2 mm for a larva at 6 days post-fertilization (dpf); Zfin) and can be easily handled using a wide bore transfer pipette or a robotic handling system see . They caA dermal bone with flat morphology and a superficial localization on the side of the head, combined with an early and rapid ossification in larval zebrafish , are keyThe operculum assay is fast (4 days from exposure to data acquisition) and requires a low quantity of molecules (exposure is short and done in a reduced volume of fish water), but also few resources that are commonly found in most research institutes or easily implemented . It is also adaptable to different experimental settings  and it is performed with early-stage larvae that can be produced in large quantities. ImageJ macros have been developed to speed up the rather laborious morphometric analysis  and toolZebrafish craniofacial bones are ossified following mechanisms that have been conserved throughout vertebrate evolution and are therefore commonly used as models for mammalian cranium development . FlemingZebrafish vertebral centrum is formed throughout intramembranous ossification that can be detected as early as 7 dpf, when part of the notochord sheath began to mineralize . QuantifTg(sp7:mCherry) or Tg(Ola.Sp7:Luciferase), where an easily quantifiable reporter signal is used as a proxy for bone formation, has accelerated the screening procedure by shortening ex vivo culture, limiting scale handling and simplifying image acquisition [Zebrafish elasmoid scales are small bone-like units of easy access that can be easily and rapidly plucked out of zebrafish skin. They are available in rather large quantities—around 200 elasmoid scales in a single adult zebrafish ,51—and cuisition . The smauisition , justifyThe ability of the zebrafish to fully restore damaged/amputated skeletal structures—e.g., fin rays, skull, jaw and scales—has been used to study molecular and cellular mechanisms underlying de novo bone formation ,62,63,64Zebrafish caudal fin is a simple structure composed of bony rays spaced with vascularized and innervated connective tissue and covered with a pigmented epidermis . Upon amWhen they are lost or removed experimentally, zebrafish elasmoid scales can fully regenerate within 2–3 weeks following a process comparable to intramembranous ossification . As for Injuries to bony rays and cranial bones in the adult zebrafish have been used to model human bone fractures and study bone repair ,71,72. IThe lower jaw of adult zebrafish is a simple skeletal structure primarily composed of the mandibular bone and Meckel’s cartilage . Upon amchihuahua mutation in col1a1 to mimic osteogenesis imperfecta [Molecular mechanisms and cell types associated with skeleton development and mineralization have been remarkably conserved throughout evolution ,74,75,76perfecta ). The deperfecta  and genoperfecta  and CRISperfecta ,81. Thesperfecta ,83,84,85perfecta  resultedperfecta .Several excellent reviews addressing the use of zebrafish mutant lines to study skeletal disorders have been published ,89,90,91casper [crystal [Besides the lines mentioned above—those developed to model skeletal disorders and assess the expression of skeletal marker genes—other lines have sparked some interest because of the technical advantages they could bring to the study and screening of bone anabolic and catabolic compounds. In this regard, casper  and crys[crystal  mutationExposure of zebrafish—mutant, transgenic or wild-type lines—to an excess of glucocorticoids, iron or dietary fat can also be used to mimic human bone metabolic disorders such as osteoporosis . OsteopoTechnological innovation applied to the standardization and mass production of the animals, to the robotic handling of animals, plates, and liquids, as well as to the automation of data acquisition and processing, can improve screening throughput and data accuracy, thus accelerating the use of zebrafish systems for bone anabolic/catabolic drug discovery B. The miWild-type zebrafish lines found in pet shops worldwide exhibit a relatively high degree of genetic diversity that triggers some variability in their phenotype, and consequently some variability in their response to compound exposure . In thisAnimals for screening purposes must be produced under conditions that ensure optimal growth and welfare to provide homogeneous and replicable data. The standards of an ‘optimal growth and welfare’ may vary considerably between wild-type and mutant lines—it largely depends on the severity of the mutant phenotype—and the term ‘most favorable growth and welfare’ is probably more appropriate. To provide optimal/most favorable conditions, housing, husbandry and feeding of the zebrafish must be adapted to all life stages and standardized. The European community directive 2010/63/EU on the protection of animals used for scientific purposes establishes general guidelines for water quality, chemical and physical parameters applicable to all fish species; however, it does not specify zebrafish requirements.Recommendations for housing and husbandry of zebrafish were recently refined , providwww.sparos.pt, accessed on 22 May 2022) produces tailor-made diets for zebrafish.The nutrition of model animals has been recognized as an important variable in research outcomes and the use of ‘standard reference diets’ has been proposed to improve the replicability of scientific data . Among tTo generate statistically meaningful results, zebrafish high-throughput screening requires the daily production of thousands of synchronized embryos . To decrease the intensive manual labor that would result from the manipulation of dozens of tanks and hundreds of breeders and increase embryo synchronization, large-scale egg production systems have been developed . CommercTg(sp7:mCherry)—can also be efficiently sorted in 96- or 384-well plates based on a fluorescent signal, although they have to be first harvested manually from the fish as plucking has not yet been automated. In a general manner, automatic sorting is efficiently applied to tissues or whole animals in the range 0.1–10 mm.Targets for drug screening in zebrafish can be cells, tissues or whole animals (embryo and larvae). While cells—adherent or in suspension—can be easily arrayed in multiwell plates using liquid-handling robots, tissues and whole animals have been typically sorted out under a stereomicroscope using wide-bore transfer pipettes, in a rather laborious process. Automation of this process can be carried out using a fluorescence-gated sorting system—e.g., the COPAS FP-1000/2000 of Union Biometrica —that hasHigher screening throughput and higher data accuracy can be achieved using liquid-handling robots to rapidly dilute and/or dispense precise amounts of the compounds from stock library plates and array targets. Robotic solutions are commercialized by different companies  and can be adapted to the zebrafish systems used to screen for osteogenic compounds. To further increase throughput screening, plate-handling robotic arms can also be used , although their implementation requires a massive investment which probably limits their usage to big Pharma and Biotech companies. If compounds have to be delivered through microinjection, robotic injection systems have been developed: perivitelline injection for embryos ,122 and To improve the throughput and quality of screening pipelines, image acquisition has to be automated and use rapid and high-resolution camera. In the case of zebrafish screening, the microscope objective should also have the capacity to capture large objects such as larvae. While examination of wild-type and mutant/diseased animals for developmental, morphological, and functional changes upon compound exposure can be done, the use of transgenic reporter fish expressing fluorescent proteins can greatly increase the automation of image acquisition. In this regard, fluorescent microscopes with automated stages  or microplate readers coupled to high resolution camera  will speed-up the examination and imaging (in the minute range) of zebrafish embryos or larvae . ImagingAutomated screening systems generate large quantities of images that need to be analysed using specialized algorithms to provide accurate data in a timely manner. Commercial solutions and open-source software are available for automated image analysis, providing macros to count, measure, characterize morphometry, and classify objects. Most of them can also be used for image acquisition—providing tools to control microscope and capture images—and all of them can be easily applied to screening pipelines using zebrafish  and drugs (safety assessment before marketing). Zebrafish assays and technological innovations described here can also be used to screen compounds for osteotoxicity .


Artificial cells are developed to redesign novel biological functions in a programmable and tunable manner. Although it aims to reconstitute living cell features and address ‘origin of life' related questions, rapid development over the years has transformed artificial cells into an engineering tool with huge potential in applied biotechnology. Although the application of artificial cells was introduced decades ago as drug carriers, applications in other sectors are relatively new and could become possible with the technological advancement that can modulate its designing principles. Artificial cells are non-living system that includes no prerequisite designing modules for their formation and therefore allow freedom of assembling desired biological machinery within a physical boundary devoid of complex contemporary living-cell counterparts. As stimuli-responsive biomimetic tools, artificial cells are programmed to sense the surrounding, recognise their target, activate its function and perform the defined task. With the advantage of their customised design, artificial cells are being studied in biosensing, drug delivery, anti-cancer therapeutics or artificial photosynthesis type fields. This mini-review highlights those advanced fields where artificial cells with a minimalistic setup are developed as user-defined custom-made microreactors, targeting to reshape our future ‘life'. Living cells are self-optimised functional units that can execute diverse processes in a closed compact system. Natural cells originated from non-living chemicals that eventually evolved to be called ‘living'. Therefore, building a living system out of non-living materials is essential to understand how life originates. Regardless, having a fair understanding of natural cell processes, developing lab-made cells or artificial living systems is still a challenging mission for scientists . HoweverOne might ask if an artificial cell is known to be non-living, then under which circumstances it might be transformed into ‘living.' Currently, there are substantial challenges to work with before making any progress towards building artificial living cells . There iHowever, there is noticeable advancement in establishing those components individually in synthetic systems. The application of lipid vesicles as cell membranes, able to encapsulate cellular ingredients, is a fast-growing field of research . Signal-Apart from the scientific interest in building a living system out of scratch and the origin of life research, a question still might arise why do we need artificial cells in modern life, or is there any unmet need that an artificial cell can solve and a living cell cannot fulfil? The answer is yes; there are many advantages and application of artificial cells that combines molecular simplicity and technological advancement.In vitro protein expression systems, also known as cell-free protein expression systems (CFPS), are often found to have advantages over in vivo systems (E. coli expression), in particular, for the expression of low-expressing transmembrane proteins and application of non-canonical amino acids . Me. Me63]. Rhodobacter sphaeroides bacteria) were reconstituted within GUV to allow the development of light-induced pH change across the synthetic cell membrane [Photosynthesis process is able to transduce light energy into chemical energy that fuels the living functions of the photosynthetic organism. Reconstituting photosynthesis inside artificial cells would be able to provide sustainable energy sources for the coupled biomimetic function inside the artificial cell. Photosynthetic reaction centre proteins (RC) are transmembrane proteins embedded in the photosynthetic cell membrane responsible to initiate the light energy transmission process . These pmembrane . This prin vitro.Light-activated ATP synthesis within an artificial organelle was made possible using two membrane proteins; a photo-converter and an ATP synthase ,67. Thisin situ ATP in a feedback loop.Autocatalytic ATP generation to supply continuous energy to the artificial cell was efficiently programmed within GUV as a combination of artificial photosynthetic organelle and CF extract for the protein synthesis . Within Carbon fixation by directly entrapping carbon dioxide was made possible inside an artificial cell. An artificial cell was designed to mimic the function of natural chloroplast by converting carbon dioxide into multicarbon compound glycolate . Light-iDesigning artificial cells can provide a platform for understanding the biological principles of natural cells; in addition, its engineering framework reveals an exciting field of study that allows its design to be customisable for many functions. Membrane-enclosed artificial cells have the advantage of designing with hand-selected biomaterials, primarily engineered genomes encoding signalling proteins or metabolic enzymes and artificial organelles that can easily target living cells, tissues, chemicals or biomolecules. However, an artificial cell design must fulfil some functional conditions to be able to use as a bioengineering tool. Artificial cells must sense the surrounding, recognise the target or migrate to the target site, activate themselves and transmit their synthesised product to initiate the required function. As a biosensor, artificial cells must interact with the surrounding environment to activate their function, which primarily depends on membrane permeability and cell–cell communication. In the current scenario, artificial cell biosensing studies are based on the small chemical exchanges through a lipid membrane that limits the artificial cell applicability as a biosensor. Although reconstitution of membrane proteins within lipid vesicles adds some flexibility to target molecular recognition, to date, artificial cell study is limited to a small number of analytes. Advancements in membrane permeability that can allow not just ions or small molecules but also selected proteins or nucleotides might improve artificial cell design as biosensors.in vivo drug targets, it becomes challenging to mimic complex cellular membranes with their simple and homogeneous lipid composition. Therefore, target-specific modification in the lipid composition is required to design an artificial cell with efficient drug delivery capability.Artificial cells have a huge importance in the drug delivery system, which depends on the efficient encapsulation property of the lipid membrane. Currently, liposome is most favoured as the membrane in drug delivery applications due to its biodegradability and its phospholipid bilayer, which is closer to the biological cell membrane. However, for in vivo studies for proper investigations. The biggest challenge in this field is that these studies are relatively new and still in their early stages of development.In therapeutic application, ongoing artificial cell research uses both membrane functionality and CF system for on-site, sustainable therapeutic protein expression. This enables using a huge variety of therapeutic molecules using the CF system in an artificial cell. Artificial cells formed with natural cell-derived membranes have an advantage over lipid membranes. With this kind of membrane, it can bypass the host's immune defence and accumulate primarily on target cells with common cell-adhesion molecules. Although this is a unique strategy of tumour targeting method, the concept needs further Artificial photosynthesis could be a life-changing research effort in the near future, which might be helpful for solar energy conversion, biofuel generation and carbon fixation. Artificial cells as portable artificial photosynthetic devices have definite potential, which is currently limited to a very small number of studies.Artificial cell design is a combination of membrane engineering and the core functional module, which needs to develop parallelly. The biggest challenge in artificial cell research is developing a customised membrane with the required functionality, permeability and compatibility so that the core functional modules can operate by effectively sensing the environment and delivering an accurate signal as a response. So far, with a handful of studies, it is difficult to explore the practical application of artificial cells fully. However, the combination of biology, medicinal application and advancement in engineering techniques might develop artificial cells to be applicable in biotechnology.Artificial cells are a non-living synthetic design developed to mimic biological functions and modulate them using a minimalistic setup. Having relatively simple engineering principles, artificial cells are equipped with a membrane and membrane encapsulated functional core.Including biosensing and drug delivery applications, artificial cells are designed and developed in various sectors as portable bioengineering devices able to communicate and respond to the surrounding.An artificial cell has huge potential in future therapeutics and synthetic biology applications, which currently lack customised membrane options and selective membrane permeability. However, progress in membrane engineering might be able to circumvent this limitation in the near future, which would accelerate artificial cell applicability in biotechnology.


Heart failure with preserved ejection fraction is an increasing challenge for modern day medicine and has been drawing more attention recently. Invasive right heart catheterization represents the mainstay for the diagnosis of diastolic dysfunction, however due to its attributable risk of an invasive procedure, other non-invasive clinical pathways are trying to approach this pathology in clinical practice. Diastolic failure is complex, and imaging is based on various parameters. In addition to transthoracic echocardiography, numerous novel imaging approaches, such as cardiac magnetic resonance imaging, computed tomography, positron emission (computed) tomography or single photon emission tomography techniques are being used to supplement deeper insights into causal pathology and might open targets for dedicated therapy options. This article provides insights into these sophisticated imaging techniques, their incremental value for the diagnosis of this poorly understood disease and recent promising results for an enhanced prognostication of outcome and therapy monitoring. Heart failure is a significant and well-known challenge in modern day medicine. Particularly the subgroup with preserved ejection fraction is underrecognized and treatment of underlying etiologies often delayed. It accounts for about 50% of heart failureWhile resuming the indisputable clinical impact of heart failure with preserved ejection fraction (HFpEF) the important question arises: why was it such an underrecognized disease with lacking attention until lately?Going back to the “older brother” HFrEF, the quantification of the ejection fraction (EF) with transthoracic echocardiography (TTE) is a straightforward method to make evidence-based assumptions on the stage and outcome of the disease.On the other hand, the diagnosis of HFpEF requires multiple steps including pretest assessments and verifying diagnostic tests as soon as it is suspected. Therapeutic consequences are rare,,In practice, two scoring systems are suggested as the most practical approaches for diagnosing HFpEF: The H2FPEFF score .,While AF is most likely a consequence to the increased LV filling pressures with subsequently increased pressures and dilatation of the atrium,Imaging in HFpEF is mainly based on visualizing the decompensation of the diastolic cardiac function and the subsequent consequences.During a regular cardiac cycle, the diastole is subdivided into major phases including: 1. relaxation of the LV and a base-ward movement of the mitral annulus to fill the LV with blood; and 2. the atrial contraction to guarantee augmentation of ventricular filling in late ventricular diastole. While the first phase during the early diastole is an active mechanism of LV myocytes and biochemical events to initiate ventricular relaxation, the second phase during late diastole is a passive property of the LV, actively contributed by the LA.Finally, during the ventricular systole, the atrial relaxation initiates a passive collection of the pulmonary venous return into the atrium to initiate another diastolic cycle.,,Those three phases are markers of a specific cardiac function as measures of compliance and contractility,,In diastolic dysfunction, impaired LV relaxation and stiffness determine impaired atrial conduit function, which induces an increase of the atrial booster pump function to compensate for the decreased LV filling. This is paralleled by a mismatch of both diastolic phases i.e., the ventricular influx by the atrial contraction catches up with the influx by the atrial conduit function. The incremental atrial workload secondary to the ventricular diastolic disfunction can result in atrial decompensation. Hand in hand with the failure goes atrial dilatation and an absolute reduction of the atrial stroke volume, leaving by implication, a reduced ventricular influx during diastole.Patients suffering from HFpEF are characterized by their inability to adequately react to physical exercise due to an impaired atrial contractile reserve.HFpEF is defined as a PCWP ≥25 mmHg during exercise,The observation of increased PCWP during exercise stress was a milestone in the diagnosis of HFpEF and allows the identification of an otherwise clinically inapparent disease for the first time.As RHC is an invasive procedure, other diagnostic pathways have been explored for an early diagnosis in clinical routine.The most frequently used pathways are based on scoring systems consisting of clinical, laboratory and echocardiographic measurements to estimate the probability of HFpEF, the H2FPEF-ScoreBoth scores include parameters such as bodyweight, aHT, age and AF as well as distinctive echocardiographic parameters.In case of uncertainty with e.g., intermediate scores functional testing with stress-echocardiography on the non-invasive side, or with RHC on the invasive side is recommended.As soon as the diagnosis is established, an etiological work-up is necessary referring to a variety of diagnostic methods such as cardiac magnetic resonance imaging (CMR), biopsies, computed tomography (CT), positron emission computed tomography (PET-CT), single photon emission tomography (SPECT) or genetic testing. Only by the use of those advanced imaging- and diagnostic methods—HFpEF-like syndromes with advanced targeted therapy options can be identified and treated.,Echocardiography must be highlighted first, as it is included in both above-mentioned scoring system and the primary approach to screen for a preserved EF. Starting with structural parameters there is the left ventricular mass index (LVMI) as a direct measure of myocardial hypertrophy with a consecutive thickening of the measured walls. The left atrial volume index (LAVI) is assessed in two orthogonal views and indicates atrial size.An enhanced echocardiographic assessment of diastolic function should include the inflow through the mitral valve (E) and the inflow due to the contraction of the atrium (A) as indirect measures of the atrial conduit and booster pump function. Furthermore, the velocity of the mitral annulus (e′) should be measured in the septal and lateral region. In addition to the individual parameters, the derived ratios  are important for the diagnosis and monitoring of HFpEF see .Fig. 2Pa,All illustrated functional parameters are measured by pulse-wave and tissue doppler signals over the mentioned structures. Those flow-derived echo parameters depend on a correct alignment of the ultrasound beam with the flow direction, which can be difficult, even for experienced echocardiographers.,,A non-angle-dependent functional equivalent and innovative method to monitor LV shortening is multidimensional global longitudinal strain (GLS) using speckle tracking compare .48 Reduc,As outlined before, exercise stress testing is an important element of the diagnosis of HFpEF and is mostly recommended in cases of uncertainty.At the moment, CMR is being recommended for the detailed etiological work-up after establishing the diagnosis using the mentioned guidelines.,As used for various other diseases already, CMR-based tissue differentiation allows for the quantification of myocardial fibrosis by calculating the extracellular volume fraction (ECV) using native and contrast-enhanced quantitative T1 mapping sequences(99m)Tc-DPD Scintigraphy for the identification of cardiac amyloidosis,For the work-up of other cardiomyopathies, guidelines recommend further enhanced diagnostic imaging. This includes Even though those imaging approaches are an indispensable part of the diagnostic work-up, their representation in current guidelines is rather small as some advances still need further validation while the increased costs compared to TTE cannot be disregarded. However, those innovative techniques using multimodal imaging show promising results for the initial diagnosis of HFpEF and are an important area of research to combine the ability of tissue characterization and diagnostic testing within one modality.Multiple non-invasive imaging techniques improve the understanding of the underlying pathophysiology in HFpEF. This paragraph aims to give a brief overview on some of those methods, and their potential use in the clinical routine compare Table 1No,,At the present time CMR represents the reference standard for the assessment of myocardial systolic function due to its inherent superiority over TTE resulting from the better endocardial surface definition and its independency from acoustic windows.Nevertheless, as previously shown, exercise stress is of great importance for the diagnosis of HFpEF. Novel real time CMR techniques allow the acquisition of individual heart beats with high resolution in real time without the need to average cardiac function over several heart beats. The proposed techniques facilitate a quantitative assessment of diastolic and systolic myocardial function using physiological exercise CMR.,Since arterial stiffness as a decrement of vessel compliance was observed to be strongly associated with echocardiographic parameters predicting diastolic dysfunctionAs CMR phase-contrast flow-measurements can be used for a comprehensive calculation of pulse-wave-velocity see  over the,By the quantification of phase shifts in a specific phase-encoding direction, CMR measures blood flow velocity and absolute volumes .82 By auIntracardiac 4D-flow measurements were able to deliver deeper insights to patients with conventional heart failure and the prediction of their exercise capacity. In particular, in patients with dilated or ischemic cardiomyopathy, the direct kinetic energy was shown to be an independent predictor of the functional capacity and could be used for therapy monitoring and prognostication in the future.In patients with atrial fibrillation despite of similar LV mass, volume and EF; 4D-flow measurements were able to indicate that patients suffering from AF had reduced direct flow and a delayed ejection compared to controls.Albeit this method needs further validation, its calculation can be performed semi-automatically with low inter- and intraobserver variability and independently from potential confounders like the angulation in TTE.While the role of the great arteries in HFpEF has been described above, the importance of coronary vessels should be equally emphasized. Coronary artery flow is known to mainly occur during diastole of the cardiac cycle. The impaired lusitropy in HFpEF may reduce coronary artery flow and subsequently myocardial perfusion, which in turn is highly likely to accelerate myocardial remodeling followed by diastolic dysfuntion.Microvascular disease can be diagnosed by a reduced coronary flow reserve with PET-CT being a well-established non-invasive test for this diagnosis. Using PET and distinct tracers, it is possible to accurately quantify myocardial blood flow and the myocardial perfusion reserve .94 As anOn the other end of coronary flow, coronary venous congestion was found to be another important player in HFpEF.,Pulmonary arterial hypertension or pulmonary oedema with subsequently increased RA filling pressure can result in coronary venous congestion, which is known to cause myocardial edema.In clinical imaging, CMR tissue characterization with T2-based imaging and mapping of myocardial edema was strongly associated with hypertrophy and diastolic dysfunction.,Cardiac edema is a marker for inflammation and can be assessed using T2-weighted imaging in CMR as explained previously. While edema is just an indirect measure of inflammation, FDG-PET can differentiate small changes in myocardial metabolism. While the atria cannot be sufficiently assessed by CMR T2 mapping, PET is able to identify focal regions in the atria with increased metabolism as a sign of inflammation.64Cu-macrin in PETMore experimental techniques enable direct imaging of the inflammation in the heart by additional use of hyperpolarized contrast agency in CMR,123I-MIBG SPECT to determine sympathetic presynaptic nerve function, a correlation between the severity of diastolic dysfunction and remodeling was found.Besides cardiac metabolism, increased cardiac innervation by the sympathetic nerve system with an increased adrenergic drive was found to accelerate cardiac remodeling.As mentioned, adipose tissue has an important role as a substrate and risk factor for diastolic dysfunction and HFpEF.Another important adjunct is the visualization of the pulmonary congestion causing exertional dyspnea as one of the first symptoms in patients with HFpEF.,Transit pulmonary congestion could be observed during submaximal exercise stress already, and was linked to impaired exercise hemodynamics in patients with HFpEF.Using proton density maps, CMR allows for a straightforward quantification of lung water density see  and qual,,HFpEF and HFrEF were observed to have similar outcomes regarding hospitalization, symptomatic burden and quality of life.Innovative techniques may pave the way for an enhanced characterization of a complex pathophysiology; however, they will have to undergo further validation and the clinical benefit has yet to be proven. Some of the techniques are very complex in their everyday application and require highly specialized centers to use and interpret them correctly. The simplification and standardization is an important area of ongoing research.In clinical routine, diagnostic markers are indispensable for the correct description of a patients’ medical condition. Further studies will need to focus on various aspects while using innovative imaging markers. This includes reproducibility, cost efficiency and efficiency of use, while they must be closely related to the pathophysiology and ease diagnostic decision making to identify patients at risk and improve their outcomes.The non-invasive diagnosis of HFpEF is challenged by the complexity of the disease in clinical practice. While modern techniques like speckle-tracking for echocardiography enhance the monitoring of the LV contraction and relaxation, other imaging modalities like CMR, CT or PET-CT are quickly evolving. By using the combined capabilities of myocardial tissue characterization and functional quantification at rest and during exercise, CMR yields high diagnostic value for the diagnosis of HFpEF. Recent developments promise more insights into the complex pathophysiology of this condition, a better risk stratification, and better therapeutic guidance.PubMed and relevant articles from 1990 to 2022 were screened for references to this article. We included the search terms: “HFpEF”, “HFpEF diagnosis”, “HFpEF imaging”, “Diastolic dysfunction”, “Heart failure with preserved ejection fraction”, “HFpEF non-invasive diagnosis” and “HFpEF pathophysiology”. The final references were chosen based on their relevance to the main topic of the article and their importance to current diagnostic guidelines and innovations. All references were discussed by the authors before their final use.Both authors wrote, reviewed, and edited this manuscript. Both authors read and approved the final version of the manuscript.None.


Low anterior resection syndrome (LARS) describes the symptoms and experiences of bowel dysfunction experienced by patients after rectal cancer surgery. LARS is a complex and multifactorial syndrome exacerbated by factors such as low anastomotic height, defunctioning of the colon and neorectum, and radiotherapy. There has recently been growing awareness and understanding regarding the role of colonic motility as a contributing mechanism for LARS. It is well established that rectosigmoid motility serves an important role in coordinating rectal filling and maintaining continence. Resection of the rectosigmoid may therefore contribute to LARS through altered distal colonic and neorectal motility. This review evaluates the role of colonic motility within the broader pathophysiology of LARS and outlines future directions of research needed to enable targeted therapy for specific LARS phenotypes. Rectal cancer prevalence is increasing, particularly in those under 50 , with suSeveral terms have been used to describe these survivorship disorders such as ‘pelvic radiotherapy induced dysfunction’, and ‘low anterior resection syndrome’. Recently, the symptoms and consequences of bowel dysfunction that patients suffer after rectal resection, termed low anterior resection syndrome (LARS), have been defined and standardised based on international consensus . LARS isIn the setting of increasingly minimally invasive surgery, chemoradiotherapy, and emerging watch and wait approaches , 15, surThe heterogenous symptoms experienced by patients with LARS likely also reflects the multifactorial pathophysiology of the syndrome. Surgical trauma to the anal sphincter complex, colonic denervation, reduced neorectal capacity and compliance, radiotherapy-induced fibrosis, and faecal diversion, have all been shown to negatively impact bowel function after rectal resection , 17–20. The term LARS has historically been used to report heterogeneous symptomatology often with a focus on faecal incontinence, neglecting other symptoms shown to be more strongly associated with patients’ postoperative quality of life . A recenAccurately measuring LARS in a standardised manner is an essential step to better understanding the pathophysiology of LARS. Historically, a wide range of validated and unvalidated tools have been used to report LARS, further contributing to the complexity of assessing this syndrome . The besCurrent management of LARS is reactive, empirical, and symptom based . Managemvia the L1-L2 splanchnic nerves (emanating from the inferior hypogastric plexus). The sympathetic nerves inhibit proximal colonic motility tonically  was observed in patients with medically-refractory faecal incontinence compared to healthy controls . While tvs right-sided resections  and 9 healthy controls . Keane eHRCM demonstrated attenuation of the rectosigmoid brake mechanism in the patients with LARS, as they had fewer retrograde propagating contractions pre- and post-meal, which propagated for a shorter distance with reduced amplitude post-meal, compared with healthy controls Figure 3Additionally, Vather et al. showed that in 9 out of 12 patients with “normal” bowel function post-anterior resection, propagating motor patterns (such as HAPCs and the CMP) traversed the anastomosis, with no drop-off at the site of the surgical scar . Return HRCM evidence has suggested that the loss of rectosigmoid brake function as well as loss of long gastrointestinal feedback loops  may be plausible mechanisms underlying LARS. These hypotheses are further supported by other physiological modalities; Mochiki et al. performed a barostat study in 37 patients after low anterior resection, 17 with high stool frequencies and 20 with normal stool frequencies and found an increased gastrocolic reflex in patients after low anterior resection . While aColonic motility is likely an important contributor to the complex and multifactorial underlying pathophysiology of LARS. Other critical aetiological mechanisms during surgery relate to disruptions to intestinal continuity, denervation of the mobilised left colon, and injury to the anal sphincter complex. Construction of a neorectum with altered motility (often secondary to nerve injury) , and decWhile risk factors for LARS after anterior resection  , 12 haveAnastomotic height has been shown to be a consistent risk factors for poor functional outcome after rectal resection –12. The in-situ may allow greater compensation after rectosigmoid resection. The distal colon has important storage functions, which may reduce ability to delay defecation when these reserves are lost along with the rectal storage capacity  and colonic J pouches (CJP) are often used in an attempt to mitigate against bowel dysfunction associated with straight or end-to-end anastomoses . FunctioDenervation of the colon and rectum contributes to the pathogenesis of LARS, and is likely a significant mediator of altered colonic motility after surgery . Surgicavia either surgical or chemical neurectomy of the IMA pedicle increases rectal motility and colonic transit, while denervation of the parasympathetic supply has the opposite effect  among patients that had LAR for rectal cancer with colorectal or coloanal anastomosis . HoweverLARS pathophysiology is receiving increased attention and this review highlights the importance of a standardised definition of LARS to guide consistent clinical investigation. Given the varying symptoms experienced, numerous mechanisms are likely to contribute to LARS, with specific symptom experiences potentially explained by overlapping factors. The role of colonic motility has been a relatively underappreciated contributor. Further work is required to better correlate symptoms of LARS with profiles of colonic dysmotility and such advances may direct targeted therapies, including SNM through its modulation of colonic motility . AdditioThe highly invasive nature of colonic manometry has limited the study of colonic motility, often resulting in small study cohorts. Emerging technologies that enable non-invasive detection of colonic myoelectrical activity through skin-surface electrical recordings have demonstrated the ability to detect and quantify meal responses . These aAltered distal colonic motility is an important contributor to the pathophysiology of LARS and may offer an actionable biomarker for targeted therapies. Further work is required to enable accessible and accurate assessment of colonic motility, to guide understanding of LARS aetiology and to facilitate physiologically congruent management strategies for these patients.IB, GO’G, and CK, provided supervision. CV, CW, IB, GO’G, and CK wrote the paper and approved the final version for publication.This work was supported by the New Zealand Health Research Council and the Royal Australasian College of Surgeons’ John Mitchell Crouch Fellowship.We would like to thank Patrick Lane for their assistance with the medical illustrations.GO’G, IB, and CK are members of The University of Auckland Spin-out companies: The Insides Company Ltd , and Alimetry Ltd (GO’G).The remaining 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.


Qualitative research with 60 males (16–80 years) from two Caribbean countries wascarried out to explore men’s perspectives on domestic violence (DV). Aninductive latent/thematic approach to data analysis supported by analyticsoftware led to five key domains being identified: (1) meanings of violence; (2)patrinormative culture; (3) normalization of violence; (4) male victimization;and (5) blame attribution and empathy. Patriarchal values, together withchildhood exposure to violence, were found to reduce empathic capacity andcontribute to the normalization of DV. In addition, the minimization of malevictimization and the lack of behavior-change support services for men wereidentified as contributory factors. Gender-based violence (GBV) is a widespread public health issue limiting the livesand livelihoods not only of women, but also of sexual minorities and transgenderpeople . The mosIn the Caribbean, violence against women is acknowledged as being extensive acrossthe region , andcommensurate rates of per capita income. This is important because low rates ofsocial development have been associated with societal attitudes that justify DVand have been said to predict rates of perpetration and victimization . In 2017iswidely acknowledged that there are regional commonalities in the manifestations ofpatriarchal assumptions and the embeddedness of structural gender inequalities that apply to both countries and the datahave therefore been merged.This article is based on a subset of data gathered between 2016 and 2018 as part of astudy by the None in Three Centre for the Global Prevention of Gender-based Violencewhich aimed to explore the social etiology of DV in Barbados and Grenada. The aim ofthe current study was to explore male perspectives on DV. Although it is not assumedthat Barbados and Grenada share the same cultural norms, it The study consisted of a secondary analysis of transcripts from focus groups withmen. Convenience, purposive, and nonprobability sampling techniques were used toidentify a diverse sample of single and ever-partnered males (16–80 years) fromthree socioeconomic groups: unemployed, manual/low-skilled workers, andprofessional/high skilled and including males who had been convicted of aDV-related offense as well as males from the general public. The study utilizeda cross-sectional qualitative design—data collected at a one-time point throughguided focus-group discussions with purposively selected participants; eightfocus groups were held, four in each country. The objectives were to ascertainthe views of men concerning DV.A total of 60 men (age 26 + ) and young adults (16–25) took part, 14 men and nineyoung adults from Barbados and 23 men and 14 young adults from Grenada. Among the sample, 11 of the participants had beenconvicted of DV-related offenses, four men and three young adults from Barbadosand two men and two young adults from Grenada. Offense details were not soughtbut all of the males who had perpetrated DV-related offenses had taken part in acourt-mandated treatment program.Prompt questions included: how men define DV; views on prevalence, causes, andeffects; how living with DV affected them; situations/circumstances thatcontribute to men being violent; views about the help that men need. Ethicalapproval for the study was granted by the University of Huddersfield, School ofHuman and Health Sciences Ethics and Integrity Committee. Briefing informationemphasized the voluntary nature of participation and the right to discontinue atany time. Participants gave informed consent and focus group participantsselected pseudonyms to protect their anonymity. All focus groups were conductedin private settings, and the need for confidentiality was stressed.Focus groups were digitally recorded, recordings transcribed and an inductivelatent/thematic approach applied to data analysis. The NVivo software program(v10)  was used for coding. Key themes werecollaboratively identified by both authors to generate a codebook to guide theiterative evaluation of all data. Text searches were carried out for contentvalidity, to identify negative cases, and to check for coding density. Given thenature of qualitative research, lexical searches cannot be reduced to theirfrequency counts and phrases were therefore analyzed in the context of thesurrounding text. A constant comparison between cases enriched analysis bypointing to negative cases where men described developing nonviolent tendenciesdespite being immersed in subcultures of violence acceptance. More complexmatrix queries based on node attributes highlighted segments of the transcriptsfor further thematic analysis. The authors jointly agreed on the selection ofquotations to best illustrate the themes.n = 11) were men and young adults who hadbeen convicted of DV-related offenses, while the majority (n = 49)were men and young adults without any DV-related convictions. The intention, incombining both groups, was to explore differences in perspectives depending onwhether participants had engaged in a psychosocial treatment program or not . Two important differences emerged. Males who had participated in atreatment program demonstrated greater awareness of the situation of women and wereless likely to blame the victims of DV than the participants from the generalpopulation. However, given the small number of offender participants(n = 11) this finding should be regarded as tentative.Concerning age-related differences, older men (26 + ) tended to subscribe to moretraditional gender stereotypes than young adults (18–25) and young adults suggesteddifferent coping strategies, such as the use of marijuana rather than alcohol. Theyounger participants also argued for youth-led counseling interventions in additionto the men-to-men approaches advocated by older males. Beyond these points, therewere no other notable differences across the groups and the data were subsequentlymerged for further analysis.Of the participants, 18% . Using a deductive approach, this involved moving betweennodes and the original transcripts to identify and code units of text and referencesbased on the taxonomy. Items that shared a semantic relationship were synthesized toproduce five key domains: (1) meanings of violence; (2) patrinormative culture; (3)normalization of violence; (4) male victimization, and (5) blame attribution andempathy see .The use of focus groups meant that it was not always possible to attribute responsesto individual men or to determine precisely how many participants shared a viewbeing expressed and for this reason, in presenting the findings, phrases indicatingconsensus rather than actual numbers are sometimes used. However, the use of NVivosoftware enabled the authors to accurately determine the frequency of recurringresponses and in many instances, to track these to the participants expressing them;where this was possible, the numbers or percentages of respondents are given.n = 59) did not identify sexualcoercion as a form of DV and referred only to physical and psychological acts ofharm. Only two respondents mentioned economic control as a form of abusealthough several men thought that women's lack of economic power made themvulnerable to abuse. More than half of the interviewees questioned the ways inwhich DV had been defined by policymakers and their comments suggested thatdominant definitions had generated a polarized view of the problem resulting inthe minimization of some forms of DV. Broader conceptualizations were called forwhich include all forms of abuse and interpersonal violence, including violenceperpetrated by women and violence against and between children.I know young men who were abusing at 13,14 years old, but we don't wantto admit that you know because you see this sibling war, boys beat theirsisters and vice versa, that is abuse, that is domesticviolence.Out of 60 participants, 98.3%  expressedviews that indicated they had grown up with cultural norms that meant boys werenot encouraged or expected to recognize, express, or manage their feelings andthis led to an inability to regulate emotions.I feel man should behave a little more better with they self. Learn howto control their anger, but eh have a lot a man they just hyper, anylittle thing does just trigger them off; … Them have to learn how tocontrol their anger.This domain yielded the largest number of references see  and indiThey [boys] are encouraged to behave in violent ways, you know. When theyspeak to their peers they would be encouraged in fact to continue [beingviolent]. A lot of times, um men, they male friends would actuallysupport them [in being violent] and encourage them to be a man because alot of times domestic violence is linked to being male, being a realman.Participants suggested that growing up in the midst of violence influenced thekind of psychological tools developed for responding to conflict and whentensions boiled over because males were expected to be in control, they oftenbehaved aggressively.This is all that I know … this is what my mother and my aunts would have toldme that I ain't gonna end up anything better, and therefore “well, hey,”self-fulfilling prophecy I going to become the very thing they claim I tobe.Society itself has influenced men that they are in charge. Society itselfdoes not teach men that women should be of an equal partner with them and …because of the way they grow, men believe they are in control and theydictate everything that a woman should and should not do.n = 15) of the participantssaid they were opposed to all forms of violence and even if they had grown upwith these cultural mores, rejected the notion that violence was a part of theiridentity as males. These men spoke about the strategies that had helped them indealing with relationship conflict; religion and the influence of positive malerole models being the most common. However, men who had witnessed DV as childrenor who had been subjected to abuse themselves often drew on those samerepertoires of violence and male dominance even as they demonstrated theiropposition to it.I’ve heard a lot of young men say “you see me, I’m not gonna beat mypartner like how my father did to my mother” and there are a lot ofyoung fellas now that have a different attitude and they try, they tryto close the gap between them, they and their partners. But somepartners are very rebellious and resilient.Set alongside these findings, 25%  expressed comments that indicated thatbehaving aggressively was a default position, a choice often made unthinkinglybecause in their communities, children, and boys, in particular, were socializedinto accepting violence as normal.“…wuh monkey see, monkey do” [referring to mimicking an action withoutunderstanding or thinking about the associatedconsequences].Many men  wereregularly exposed to DV in the home while 40.6% had been directly subjected tophysical violence (One of the most consistent findings in the literature is that being socialized in ahome where DV or child abuse is perpetrated increases the likelihood ofvictimization or battering in adulthood (violence . These fviolence , 2015b. violence .Many of the participants drew on repertoires of misogyny to excuse male violence.Negative attitudes to females have long been associated with gender inequality andit is argued that the family is the primary vehicle through which such attitudes aretransmitted and where patterns of violence become established and rationalizationsfor gender-based violence formulated . The curDespite being exposed to similar levels of family and community violence as theirpeers, many men rejected culturally inscribed patrinormative values that requirethem to be the holders of power and control within a relationship. And neither werethey willing to view the woman's status as derivative or subservient. Instead, thesemen had incorporated values of nurturance, compassion, and nonviolence into theirbelief systems. A common factor in these cases was the existence within their lives,of other nonviolent men and also, older men in the community who did not dismisstheir concerns and relationship conflicts, but who guided and supported them.Indeed, they recommended that male-to-male interventions, through which menchallenge anti-woman discourses and help to develop nonviolent problem-solvingstrategies could be an important strategy for addressing DV. Exploring the value ofprograms for men who commit DV is beyond the scope of this article nevertheless itappears that the most successful batterer intervention programs are those that, inaddition to prioritizing victim safety, hold men accountable and enable thereflexive deconstruction of masculine constructs that reify control and domination.An important factor affecting the reproduction of violence is its normalization andthe social processes that contribute to its acceptance. These processes include thesocial construction of masculinized identities which, developed in the context ofthe family and affirmed by peers and others, shape the attitudes, perspectives, andbehavior of young men . In her do commit acts ofviolence against their partners. In environments in which interpersonal violence is considered a regular, everydayaspect of life, females can also become socialized toward its acceptance, and notonly as victims. A strong theme from the current research was thatfemale-perpetrated DV was not taken seriously by professionals and that views aboutmale dominance made it difficult for men to report experiences of victimhood or toobtain support. Notwithstanding the cultural challenges male victims faced indisclosing abuse perpetrated by females, they believed the disparity in professionalresponses reflected gender priorities skewed against men. Although the reportedrates of DV against males may be low, women A key finding of the current study was the attribution of blame to victims and thefailure of perpetrators to take responsibility for their actions and related tothis, the lack of empathy. Many participants offered up victim-blaming narratives tojustify DV and in such instances, victims were regarded as the arbiters of their ownabuse. This is reflective of other studies that show that norms related to thejustification of violence against women are predictive of its prevalence . Within This study is limited in several respects, most notably the fact that the focusgroups were not a representative sample of men and young adults, and in includingmales who had participated in a treatment program, the insights and behavioralchanges derived may have influenced the findings in ways that were not reflective ofthe overall sample. Future research should address these limitations. There is alsoa need to explore factors that enable men to desist from violence even when theyhave been socialized in contexts in which DV was normative. A further limitation isthat the data from the countries and age groups were merged and it was not possibletherefore to investigate country-specific or age-specific differences.The intergenerational transmission of negative gender attitudes andviolence-affirming behaviors as contributing to its normalization andacceptance.The coterminous layering of violence, meaning that violence within thehome is fuelled by factors outside it and vice versa.DV is informed by culturally/historically inscribed patriarchalconstructs which are bolstered by the structural location of females inrelation to male status and privilege.The need to engage boys, young adults, and men in violenceprevention.Despite its limitations, the study yields important insights and shows that DV is notonly sustained through gender inequalities and intergenerational processes that aregiven shape within domestic spheres but also through social, cultural, andstructural factors in the external environment. Through this coterminous layering ofviolence reproduction, negative gender attitudes are normalized and behaviors thatcontribute to DV bolstered. Also, the findings indicate that though rates ofviolence against women in the Caribbean are said to be among the highest in theworld ; 2017,i


This paper examines the anti-mask and anti-lockdown online movement in connection to the COVID-19 pandemic. To combat the spread of the coronavirus, health officials around the world urged and/or mandated citizens to wear facemasks and adopt physical distancing measures. These health policies and guidelines have become highly politicized in some parts of the world, often discussed in association with freedom of choice and independence. We downloaded references to the anti-mask and anti-lockdown social media posts using 24 search terms. From a total of 4209 social media posts, the researchers manually filtered the explicit visual and textual content that is related to discussions of different genders. We used multimodal discourse analysis (MDM) which analyzes diverse modes of communicative texts and images and focuses on appeals to emotions and reasoning. Using the MDM approach, we analysed posts taken from Facebook and Instagram from active anti-mask and anti-lockdown users, and we identified three main discourses around the gendered discussion of the anti-mask movement including hypermasculine, sexist and pejorative portrayals of “Karen”, and appropriating freedom and feminism discourses. A better understanding of how social media users evoke gendered discourses to spread anti-mask and anti-lockdown messages can help researchers identify differing reactions toward pandemic measures. Increasing evidence of the utility of face masks in reducing coronavirus transmission has normalized their use as an effective protective measure globally. However, scores of people choose not to wear a mask in public, stating different reasons for this decision. On August 29, 2020, for example, hundreds rallied in Paris to march against the newly introduced mask mandate  as it shows weakness  which is an extension of discourse analysis that uses the study of language in combination with images, scientific symbolism, gesture, music and sounds ”  engage  who are presented as aggressively complaining about anti-maskers. Men are not “Karens” even though they make similar complaints, denoting the sexist dimension in this pejorative term. Though it is difficult to know the identity of online users only from their public profiles, we found social media evidence based on users’ profiles and posts, suggesting that such pejorative labels often come from both men and women users mostly due to ideological reasons.This discourse includes posts mentioning women who project toughness, defiance, humour along with a tendency to reject rules including the use of unusual anti-lockdown measures. There is also a clear connection between anti-mask or anti-lockdown views and conspiracy theories by appropriating feminism and freedom of expression issues. Figure Interestingly, many women users in this community are appropriating popular terms from feminist movements around abortion rights, sexual freedom, and reproductive rights like #wedonotconsent, #mybodymychoice, #itsourbody, #itsourbodyitsourright, and #choice see Fig. . One FacOur results both confirm and advance previous findings regarding gender norms and public health compliance. Some studies referenced how men, more than women, agree that wearing a face covering is shameful, not cool, a sign of weakness, and a stigma; and these gender differences also mediate intentions to wear a face covering (Zanin et al., The analysis of pejorative portrayals of “Karens” demonstrates how values and beliefs around gender influence social norms regarding masking. Similar to Bhasin et al.  analysisThis paper has many limitations. First, our study focused on two social media platforms, and it does not claim to fully explore the gendered dimensions of the anti-mask and/or anti-lockdown movement because there are different other sites where this online community interacts such as YouTube, Twitter, Reddit, and Telegram. Second, we examined a limited number of visual and/or textual social media posts that explicitly feature and/or mention different genders as there could be other indirect or implicit references to gender that we could have missed. In addition, we only analysed English language posts that were overwhelmingly dealing with the US context; hence, there is a need to further understand the gendered dimension of this online community in other countries and languages. A very important point to make for possible future studies is to focus on more diverse possibilities that envision global content rather than the one which is heavily centred around North American discourses by using different search terms that take into account language differences. This may allow for a manifold audience and a wholesome dialogue around the same topic. Finally, our methodological approach is limited to identifying major discourses and could have some potential bias in terms of overlooking minor issues, so using other communication research methods like content analysis can be useful.Studying the gendered discussion around COVID-19 and anti-pandemic measures can assist in better understanding the nature of the communities that engage in such discourses on social media sites like Facebook and Instagram. For example, the relationship found here between hypermasculinity and anti-mask/anti-lockdown discourse is particularly concerning, and suggests the need for more research in this area to further examine those users propagating such ideas and others susceptible to such messaging. There is also evidence that many anti-mask users belong to conservative circles, for they seem to be motivated by an anti-liberal political agenda. The continued sexist portrayal of “Karen” across the spectrum suggests this trope is unhealthy and unhelpful. Understanding the ways social media users evoke gendered discourses to spread anti-mask and anti-lockdown messages can also help researchers gain important insight into the differences between men and women who often attack others that abide by pandemic measures.


Whilst the wellbeing of law enforcement personnel has been widely researched, border security personnel as a discrete group appear to be far less discussed, despite their roles frequently exposing them to potentially traumatic and challenging events such as contact with criminals or witnessing personal tragedies due to trafficking or smuggling.This scoping review aimed to explore existing literature to better understand the mental health of border security personnel and the factors affecting their wellbeing.Four electronic databases and grey literature were systematically searched for studies relevant to the review’s aims. Following the extraction of relevant data from each study, thematic analysis was used to synthesize findings.Thirteen studies included relevant data and were included in the review, identifying stressors including poor management; fatigue; negative public attitudes; inadequate staffing levels, resources, and training; poor opportunities for promotion; low pay; work overload; dangerous working environments; and work-related moral dilemmas.The review found that there has been little academic attention paid to border security personnel as a specific branch of law enforcement. Many of the stressors identified in this review are those also reported by law enforcement generally, although negative attitudes from the public and exposure to moral dilemmas appear to be more relevant for border security staff. Directly addressing work-related stress  may be useful in enhancing the resilience of border security personnel. This scoping review revealed that there is little literature focusing specifically on the mental health of border security personnel despite this occupational group facing unique stressors. The review identified various occupational stressors faced by border security staff; moral injury and negative public attitudes appeared to be prevalent and worthy of more research. Fostering supportive organizational cultures may enhance the resilience of border security personnel. Law enforcement personnel frequently experience mental ill-health such as post-traumatic stress disorder and depression.Law enforcement staff’s mental health appears to be associated with many occupational and organisational factors, including high demands, workload, work-family conflict, organizational injustice, low autonomy, low reward, inadequate training, inadequate resources, poor support, and traumatic exposure.Border security personnel are a distinct branch of law enforcement who have rarely been studied in isolation from other law enforcement personnel.There is little research examining the prevalence of stress or mental ill-health in border security personnel, although some evidence suggests their mental health may be poorer than the general population and other law enforcement branches.Key stressors appear to be similar to those reported by law enforcement generally, although negative perceptions from the public appear to be a prominent stressor for border security personnel despite rarely being discussed in law enforcement literature.There is little research on help-seeking in border security personnel or interventions that may help improve their wellbeing.More research specifically focusing on border security personnel should be carried out, to better understand the similarities and differences between this sub-group and law enforcement more generally in terms of their wellbeing and risk factors for poor mental health.The relationship between staff wellbeing and the perceived public image of the organization may be an important area to consider.Encouraging a supportive organizational culture, reducing mental health stigma in the workplace, and encouraging help-seeking may improve mental health outcomes for border security personnel; increasing opportunities for career development, improving the organization’s public image, reducing individual workloads, and improving the resources and training available to staff could help foster resilience.Border security personnel are a group of law enforcement personnel responsible for protecting borders and regulating the movement of goods and individuals across land, air and maritime borders. They perform a wide range of duties and are frequently exposed to trauma, tragedy, and crime—for example, facing direct threats from criminals and frustrated passengers, and having to contend with refugees (including children) who risk drowning trying to enter the country in boats, as well as refugees who risk suffocation in lorries. The role of a border security officer is characterized by the need to remain calm in stressful situations  and be cLaw enforcement personnel have been frequently identified as an occupational group at risk for mental health problems  includinLiterature reviews have established various factors associated with the mental health of law enforcement personnel. Many are work-related, including high demands, heavy workload, work-family conflict, perceived organizational injustice, lack of autonomy, lack of reward, inadequate training and inadequate equipment , 9–11. OIt is possible that border security personnel have been included in existing research, but most studies do not differentiate between their data and that of other law enforcement staff . AlthougWhat is known about the mental health and wellbeing of border security personnel?As the literature on border security personnel has never been synthesized before and it was unclear how many studies we would find, a scoping review methodology was selected as such reviews are typically broader in scope and inclusion criteria than systematic reviews. This review followed Arksey and O’Malley’s  scoping n = 1 .The first author searched PsycInfo, Medline, Embase and Global Health databases using a combination of search terms relating to border security and wellbeing; the full search strategy is available in All citations were imported into EndNote reference management software, where one author (SKB) reviewed titles, abstracts and full texts for relevance. Any clearly not meeting the inclusion criteria were excluded and any uncertainties about including a study were discussed with the second author. Relevant data from included studies were extracted onto a pre-designed spreadsheet with the following headings: authors, year of publication, country, type of publication, number of participants, job role of participants, socio-demographic characteristics of participants, design, variables measured, instruments used, key results and limitations. Thematic analysis  was usedNo ethical approval was required for this study as no primary data was collected.A total of 96 citations were found via database searching: 71 were excluded based on title, 16 based on abstract and one based on full text. One study with a potentially relevant abstract could not be fully screened due to the article being unavailable online. Another six studies were located via grey literature searching or hand-searching of reference lists, resulting in a total of thirteen studies for inclusion.Ten included studies were journal articles , 16–23, The first theme  revealed that very few studies assessed the prevalence of mental health problems in border security personnel. Two studies reported that border security police had a high prevalence of experiencing potentially traumatic events , 16, butThe third theme identified was ‘factors associated with mental health/wellbeing’. Within this theme, there was some evidence that poorer wellbeing was associated with work-related factors such as high workload, lack of control at work and poor perceived rewards . LimitedThe fourth theme (‘causes of stress’) found several stressors reported by border security personnel. These included inadequate sleep and rest , 20 and Participants reported various ‘moral challenges’ such as performing tasks they did not agree with  and the n = 168) of participants would feel uneasy about their colleagues knowing about them seeking psychological support [The theme ‘attitudes towards psychological help’ revealed that over a third of n = 8 of part support .The final theme, ‘coping’, revealed that the coping strategies most often employed by border security personnel included trying to remain positive and taking with colleagues about problems , while tThis scoping review of literature on the mental health and wellbeing of personnel working in border security described thirteen articles. We aimed to explore how much literature exists looking specifically at border security staff and synthesize the existing literature to provide an understanding of the prevalence of mental ill-health in this population and factors associated with their wellbeing. There was little research on the prevalence of stress or mental ill-health in border personnel although two studies suggested their mental health might be poorer than the general population. Whilst participants in many studies reported being exposed to trauma, there has been little investigation into the impact of traumatic exposure on border security personnel. One study suggested high workload, poor control at work and poor rewards were associated with stress. When asked to describe their main causes of stress, participants reported several different stressors relating to their roles, management and organizational culture. There was very little literature relating to help-seeking or interventions. One study found evidence that border security personnel may be reluctant to seek psychological help, whilst another found that personnel who provide peer support to colleagues may be at risk of vicarious traumatization. Participants described various coping strategies such as exercise and positive thinking, and one study (described in two papers) suggested yoga might help improve sleep and reduce anxiety.This review fills a gap in the literature by including only studies which presented data specifically on border security personnel. However, the review has limitations which means results must be interpreted with caution. First, it is difficult to synthesize literature on border security personnel from different countries because the roles and responsibilities of border security differ between countries; the levels of threat and danger faced by border staff as well as the conditions and terrain they work in are likely to differ by country. Results from studies focusing on one country may therefore not be generalizable to others, although many similarities were found in this review. Second, only a small number of articles were found, again suggesting the results may not be generalizable to the wider population of border security personnel. The finding of only a small number of articles appears to reflect a gap in the literature in terms of studying the mental health and wellbeing of border security staff. In addition, whilst the inclusion of grey literature provided us with several important studies, it must be noted that these would not have been subjected to the same rigorous peer-review process as published articles. Finally, a limitation of the review process itself is that only one author carried out the screening process; ideally, all citations would have been double-screened in order to ensure that no relevant studies were excluded.The key stressors for border security personnel appear to be poor management; fatigue and lack of adequate rest; negative attitudes from the public; inadequate staffing levels; lack of resources; inadequate training; lack of opportunities for promotion; low pay; work overload; working in dangerous environments and experiencing moral dilemmas. Many of these stressors are those also reported by law enforcement generally , 4, 9–11This review’s findings have several implications. First, and perhaps most importantly, more research focusing specifically on border security personnel is needed because currently, very little research on this topic exists. Additional research investigating factors affecting the mental health of border security personnel would allow a greater understanding of the stressors this group face and how these stressors might impact their psychological health which in turn could lead to a better understanding of how best to support this occupational group and how to prevent mental health problems. Further research on the relationship between staff wellbeing and the perceived public image of their organization—for both border security personnel and law enforcement more generally—would also be beneficial, as this review provides some data to suggest that public image might be an important factor affecting staff wellbeing but given the paucity of research examining this, replication of this finding is needed. In addition, given we found some evidence that moral challenges were a stressor for border security personnel, further research exploring potential moral injuries in this population would also be useful. The challenges of having to uphold the law and regulations whilst dealing with human tragedy and suffering amongst those who seek to enter countries illegally could potentially result in moral injury and the topic is deserving of further exploration.Despite the limited amount of data relating specifically to border security, several potential recommendations can be made. First, it is notable that several reported stressors related to managers ; many other work-related stressors also related to issues that managers may have some control over . Therefore, managers could positively affect the levels of stress in their staff by addressing these issues as much as possible. A systematic review that explored the impact of leader behaviours on employee wellbeing found that managerial support, empowerment, and positive relationships between managers and employees had a positive impact on employees’ wellbeing and ability to cope with stress . In addiInterventions that directly address work-related stress, for example by providing training in resilience and coping strategies, may help border security personnel manage their stress: for example, a systematic review of interventions for anxiety, PTSD, and fatigue in law enforcement  reportedImproving organizational culture, public image and opportunities for career advancement may also make the role of border security officer more attractive and encourage more people to pursue this career. This could result in increased staff numbers; in other words, incentives for hiring and retaining staff could help to ensure sufficient manpower, allowing officers to share their workload and have adequate sleep and rest and reduce work overload .Overall, this study found that the risk factors for poor mental health in border security personnel appear to be similar to those for law enforcement generally. Therefore, interventions and workplace policies which have been found to improve mental health in law enforcement may be similarly effective in improving the wellbeing of border security officers, although their effectiveness with this sub-group would need to be tested and evaluated.kqac108_suppl_Supplementary_MaterialClick here for additional data file.


On an environmental level, decades of research on EAMs and the derived subject termed “electromicrobiology” provide a rich collection of multidisciplinary knowledge and establish various bioelectrochemical designs for the development of environmental biotechnology. Recent advances suggest that EAMs actually make greater differences on a larger scale, and the metabolism of microbial community and ecological interactions between microbes play a great role in bioremediation processes. In this perspective, we propose the concept of microbial electron transfer network (METN) that demonstrates the “species-to-species” interactions further and discuss several key questions ranging from cellular modification to microbiome construction. Future research directions including metabolic flux regulation and microbes–materials interactions are also highlighted to advance understanding of METN for the development of next-generation environmental biotechnology.The overlap of microbiology and electrochemistry provides plenty of opportunities for a deeper understanding of the redox biogeochemical cycle of natural-abundant elements  on Earth. The electroactive microorganisms (EAMs) mediate electron flows outward the cytomembrane  After a rapid development in the past decade, scientists have contributed worthwhile endeavors to establish the scientific basis of potential microbial electrochemical technologies such as microbial fuel cells or microbial electrolytic cells to deal with various environmental issues . Up to tt cycles . For inst cycles . Furthert cycles . Beyond t cycles . In partt cycles . On this2 or acetate as electron donors for methanogenesis processes, direct interspecies electron transfer (DIET) is recognized as a highly efficient and stable process connecting both respiratory and fermentative bacteria/archaea for bioconversion from (macro-molecular) organics to methane  while the other, Methanosarcina barkeri, conducted CO2-to-CH4 conversion powered by the bio-electron flow. The key question was how those bio-electrons passed through cytomembranes. Revealed by metatranscriptomic analyses, both multihaem cytochrome c and nanofilaments (direct contact) and electron shuttles (indirect connect) wired two species to construct a biological hybrid system  and cytochromes or artificial materials (hydrochar) were evident as available electric conduits for DIET . The answer is obvious. New scientific discoveries and insights have expanded upon new electroactive species and novel electric bridges; however, a coherent and comprehensive picture or framework of METN on how it works in biological applications and its associated environmental implications is still not available. It would be of great significance to formulate this concept to guide the construction and operation of biological treatments from lab-scale to real application, in a multidiscipline view.Now, back to the question of whether METN is important for future environmental biotechnology. More than 400 scientific papers per year have been published within the broad scope of ‘‘Science. The Shewanella sp., a famous EAM model strain, was in vivo embedded with silver nanoparticles for excellent fuel-utilization efficiency in microbial fuel cells  to microbial secreta   . OverallGeobacter spp.) would largely alter the electron transfer networks in complicated microbial consortia  was constructed following a “division-of-labor” principle, resulting in better bioenergy generation during which the production of electron donors/shuttles and bioenergy recovery were separately allocated in the three-species microbial consortium  could be used to further metabolize the chromate-reducing bacteria (like Geobacter spp. or Desulfovibrio spp.). This finding was quite important as it provided a potential microbial consortia design for Cr(VI) removal in groundwater and other water streams where proper organic electron donors are insufficient and any treatments that potentially bring secondary pollution are strictly forbidden. Though promising, the scientific community may expect a database-like toolbox recording interspecific synergy or even competition to better advise the research and application attempts of the synthetic METN microcosm. Ignoring immense technology transfer issues, it is not surprising to expect that the METN effectiveness could be largely improved and dynamically controlled by means of constructing synthetic microbial consortia with designed objectives.Though great strides have been made, an ideal niche for METN remains difficult to maintain in real biotreatments. It was found that the interspecific competition even within the same genus  in METN to avoid unwanted selection bias is a huge challenge. To answer this, cytocompatible EET circuits and eco-compatible strains should be carefully constructed, selected, and assembled in the METN microcosm. Though most proposals of “cytocompatible EET” establishment are based on the modifications of electroactive species, the unique EET capability of EAMs may place themselves as potential chassis cells for “eco-compatible strains” construction to deeply modify the metabolic flux  in such biotechnology (synthetic biology). In this case, biosafety should be carefully noted in the research to avoid the diffusion of modified genes into the natural gene pool. Nevertheless, intensive “Design–Build–Test–Learn cycles” should be both essentially and iteratively conducted for objective-driven and precise optimization. Strategies including competition-related elements  and cross-feeding can be applied to maintain the balance of this synthetic METN microcosm. Though it seems impossible to universally know the specific functions of each microorganism, piecing together clues behind the mechanisms of METN would be a potential shortcut. Speaking of which, quorum sensing (QS) could be probably taken into consideration on the microbial compatibility control. QS is basically an interspecies communication process during microbial aggregation, biofilm formation, and granulation, induced by a series of QS signals like homoserine lactones and autoinducing peptides . Of mostS. oneidensis MR-1 in porous media (Microbial electron transfer network is clearly not an exclusive concept of EAMs or other microorganisms; more innovative research on the microbes–materials interactions should be conducted both technically and economically to increase the knowledge base and the competitiveness of related environmental biotechnologies. In particular, questions on how to expand the influence range of METN and to what extent could METN be domesticated still perplex environmental researchers. Energy taxis, as a key branch of chemotaxis, was recently proposed to control the transport and motility of us media . It is ius media . Attemptus media . On the us media . We are The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.SZ and DS organized ideas and drafted the manuscript. J-DG, YY, and MX provide significant advice for the manuscript and revised the manuscript. All authors contributed to the article and approved the submitted version.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.