Variation of terpene alkaloids in Daphniphyllum macropodum across plants and tissues.

Daphniphyllum macropodum produces alkaloids that are structurally complex with polycyclic, stereochemically rich carbon skeletons. Understanding how these compounds are formed by the plant may enable exploration of their biological function and bioactivities. We employed multiple metabolomics techniques, including a workflow to annotate compounds in the absence of standards, to compare alkaloid content across plants and tissues. Different alkaloid structural types were found to have distinct distributions between genotypes, between tissues and within tissues. Alkaloid structural types also showed different isotope labelling enrichments that matched their biosynthetic relationships. The work suggests that mevalonate derived 30-carbon alkaloids are formed in the phloem region before their conversion to 22-carbon alkaloids which accumulate in the epidermis. This sets the stage for further investigation into the biosynthetic pathway.

Why did Rubens add a parrot to Titian's The Fall of Man? A pictorial manipulation of joint attention.

Almost 400 years ago, Rubens copied Titian's The Fall of Man, albeit with important changes. Rubens altered Titian's original composition in numerous ways, including by changing the gaze directions of the depicted characters and adding a striking red parrot to the painting. Here, we quantify the impact of Rubens's choices on the viewer's gaze behavior. We displayed digital copies of Rubens's and Titian's artworks-as well as a version of Rubens's painting with the parrot digitally removed-on a computer screen while recording the eye movements produced by observers during free visual exploration of each image. To assess the effects of Rubens's changes to Titian's composition, we directly compared multiple gaze parameters across the different images. We found that participants gazed at Eve's face more frequently in Rubens's painting than in Titian's. In addition, gaze positions were more tightly focused for the former than for the latter, consistent with different allocations of viewer interest. We also investigated how gaze fixation on Eve's face affected the perceptual visibility of the parrot in Rubens's composition and how the parrot's presence versus its absence impacted gaze dynamics. Taken together, our results demonstrate that Rubens's critical deviations from Titian's painting have powerful effects on viewers' oculomotor behavior.

Developing a clinical decision support system software prototype that assists in the management of patients with self-harm in the emergency department: protocol of the PERMANENS project.

BACKGROUND: Self-harm presents a significant public health challenge. Emergency departments (EDs) are crucial healthcare settings in managing self-harm, but clinician uncertainty in risk assessment may contribute to ineffective care. Clinical Decision Support Systems (CDSSs) show promise in enhancing care processes, but their effective implementation in self-harm management remains unexplored. METHODS: PERMANENS comprises a combination of methodologies and study designs aimed at developing a CDSS prototype that assists clinicians in the personalized assessment and management of ED patients presenting with self-harm. Ensemble prediction models will be constructed by applying machine learning techniques on electronic registry data from four sites, i.e., Catalonia (Spain), Ireland, Norway, and Sweden. These models will predict key adverse outcomes including self-harm repetition, suicide, premature death, and lack of post-discharge care. Available registry data include routinely collected electronic health record data, mortality data, and administrative data, and will be harmonized using the OMOP Common Data Model, ensuring consistency in terminologies, vocabularies and coding schemes. A clinical knowledge base of effective suicide prevention interventions will be developed rooted in a systematic review of clinical practice guidelines, including quality assessment of guidelines using the AGREE II tool. The CDSS software prototype will include a backend that integrates the prediction models and the clinical knowledge base to enable accurate patient risk stratification and subsequent intervention allocation. The CDSS frontend will enable personalized risk assessment and will provide tailored treatment plans, following a tiered evidence-based approach. Implementation research will ensure the CDSS' practical functionality and feasibility, and will include periodic meetings with user-advisory groups, mixed-methods research to identify currently unmet needs in self-harm risk assessment, and small-scale usability testing of the CDSS prototype software. DISCUSSION: Through the development of the proposed CDSS software prototype, PERMANENS aims to standardize care, enhance clinician confidence, improve patient satisfaction, and increase treatment compliance. The routine integration of CDSS for self-harm risk assessment within healthcare systems holds significant potential in effectively reducing suicide mortality rates by facilitating personalized and timely delivery of effective interventions on a large scale for individuals at risk of suicide.

Minimal reporting guideline for research involving eye tracking (2023 edition).

A guideline is proposed that comprises the minimum items to be reported in research studies involving an eye tracker and human or non-human primate participant(s). This guideline was developed over a 3-year period using a consensus-based process via an open invitation to the international eye tracking community. This guideline will be reviewed at maximum intervals of 4 years.

In Vivo Fiber-Coupled Pre-Clinical Confocal Laser-scanning Endomicroscopy (pCLE) of Hippocampal Capillaries in Awake Mice.

The goal of this protocol is to describe fiber-optic-bundle-coupled pre-clinical confocal laser-scanning endomicroscopy (pCLE) in its specific application to elucidate capillary blood flow effects during seizures, driven by mural cells. In vitro and in vivo cortical imaging have shown that capillary constrictions driven by pericytes can result from functional local neural activity, as well as from drug application, in healthy animals. Here, a protocol is presented on how to use pCLE to determine the role of microvascular dynamics in neural degeneration in epilepsy, at any tissue depth (specifically in the hippocampus). We describe a head restraint technique that has been adapted to record pCLE in awake animals, to address potential side-effects of anesthetics on neural activity. Using these methods, electrophysiological and imaging recordings can be conducted over several hours in deep neural structures of the brain.

Eye tracking: empirical foundations for a minimal reporting guideline.

In this paper, we present a review of how the various aspects of any study using an eye tracker (such as the instrument, methodology, environment, participant, etc.) affect the quality of the recorded eye-tracking data and the obtained eye-movement and gaze measures. We take this review to represent the empirical foundation for reporting guidelines of any study involving an eye tracker. We compare this empirical foundation to five existing reporting guidelines and to a database of 207 published eye-tracking studies. We find that reporting guidelines vary substantially and do not match with actual reporting practices. We end by deriving a minimal, flexible reporting guideline based on empirical research (Section "An empirically based minimal reporting guideline").

What the Neuroscience and Psychology of Magic Reveal about Misinformation.

When we believe misinformation, we have succumbed to an illusion: our perception or interpretation of the world does not match reality. We often trust misinformation for reasons that are unrelated to an objective, critical interpretation of the available data: Key facts go unnoticed or unreported. Overwhelming information prevents the formulation of alternative explanations. Statements become more believable every time they are repeated. Events are reframed or given "spin" to mislead audiences. In magic shows, illusionists apply similar techniques to convince spectators that false and even seemingly impossible events have happened. Yet, many magicians are "honest liars," asking audiences to suspend their disbelief only during the performance, for the sole purpose of entertainment. Magic misdirection has been studied in the lab for over a century. Psychological research has sought to understand magic from a scientific perspective and to apply the tools of magic to the understanding of cognitive and perceptual processes. More recently, neuroscientific investigations have also explored the relationship between magic illusions and their underlying brain mechanisms. We propose that the insights gained from such studies can be applied to understanding the prevalence and success of misinformation. Here, we review some of the common factors in how people experience magic during a performance and are subject to misinformation in their daily lives. Considering these factors will be important in reducing misinformation and encouraging critical thinking in society.

Advancing Research on Medical Image Perception by Strengthening Multidisciplinary Collaboration.

Medical image interpretation is central to detecting, diagnosing, and staging cancer and many other disorders. At a time when medical imaging is being transformed by digital technologies and artificial intelligence, understanding the basic perceptual and cognitive processes underlying medical image interpretation is vital for increasing diagnosticians' accuracy and performance, improving patient outcomes, and reducing diagnostician burnout. Medical image perception remains substantially understudied. In September 2019, the National Cancer Institute convened a multidisciplinary panel of radiologists and pathologists together with researchers working in medical image perception and adjacent fields of cognition and perception for the "Cognition and Medical Image Perception Think Tank." The Think Tank's key objectives were to identify critical unsolved problems related to visual perception in pathology and radiology from the perspective of diagnosticians, discuss how these clinically relevant questions could be addressed through cognitive and perception research, identify barriers and solutions for transdisciplinary collaborations, define ways to elevate the profile of cognition and perception research within the medical image community, determine the greatest needs to advance medical image perception, and outline future goals and strategies to evaluate progress. The Think Tank emphasized diagnosticians' perspectives as the crucial starting point for medical image perception research, with diagnosticians describing their interpretation process and identifying perceptual and cognitive problems that arise. This article reports the deliberations of the Think Tank participants to address these objectives and highlight opportunities to expand research on medical image perception.

Mandating Limits on Workload, Duty, and Speed in Radiology.

Research has not yet quantified the effects of workload or duty hours on the accuracy of radiologists. With the exception of a brief reduction in imaging studies during the 2020 peak of the COVID-19 pandemic, the workload of radiologists in the United States has seen relentless growth in recent years. One concern is that this increased demand could lead to reduced accuracy. Behavioral studies in species ranging from insects to humans have shown that decision speed is inversely correlated to decision accuracy. A potential solution is to institute workload and duty limits to optimize radiologist performance and patient safety. The concern, however, is that any prescribed mandated limits would be arbitrary and thus no more advantageous than allowing radiologists to self-regulate. Specific studies have been proposed to determine whether limits reduce error, and if so, to provide a principled basis for such limits. This could determine the precise susceptibility of individual radiologists to medical error as a function of speed during image viewing, the maximum number of studies that could be read during a work shift, and the appropriate shift duration as a function of time of day. Before principled recommendations for restrictions are made, however, it is important to understand how radiologists function both optimally and at the margins of adequate performance. This study examines the relationship between interpretation speed and error rates in radiology, the potential influence of artificial intelligence on reading speed and error rates, and the possible outcomes of imposed limits on both caseload and duty hours. This review concludes that the scientific evidence needed to make meaningful rules is lacking and notes that regulating workloads without scientific principles can be more harmful than not regulating at all.

Perceptual hue, lightness, and chroma are represented in a multidimensional functional anatomical map in macaque V1.

Humans perceive millions of colors along three dimensions of color space: hue, lightness, and chroma. A major gap in knowledge is where the brain represents these specific dimensions in cortex, and how they relate to each other. Previous studies have shown that brain areas V4 and the posterior inferotemporal cortex (PIT) are central to computing color dimensions. To determine the contribution of V1 to setting up these downstream processing mechanisms, we studied cortical color responses in macaques-who share color vision mechanisms with humans. We used two-photon calcium imaging at both meso- and micro-scales and found that hue and lightness are laid out in orthogonal directions on the cortical map, with chroma represented by the strength of neuronal responses, as previously shown in PIT. These findings suggest that the earliest cortical stages of vision determine the three primary dimensions of human color perception.

Visual Illusions in Radiology: Untrue Perceptions in Medical Images and Their Implications for Diagnostic Accuracy.

Errors in radiologic interpretation are largely the result of failures of perception. This remains true despite the increasing use of computer-aided detection and diagnosis. We surveyed the literature on visual illusions during the viewing of radiologic images. Misperception of anatomical structures is a potential cause of error that can lead to patient harm if disease is seen when none is present. However, visual illusions can also help enhance the ability of radiologists to detect and characterize abnormalities. Indeed, radiologists have learned to exploit certain perceptual biases in diagnostic findings and as training tools. We propose that further detailed study of radiologic illusions would help clarify the mechanisms underlying radiologic performance and provide additional heuristics to improve radiologist training and reduce medical error.

Gaze mechanisms enabling the detection of faint stars in the night sky.

For millennia, people have used "averted vision" to improve their detection of faint celestial objects, a technique first documented around 325 BCE. Yet, no studies have assessed gaze location during averted vision to determine what pattern best facilitates perception. Here, we characterized averted vision while recording eye-positions of dark-adapted human participants, for the first time. We simulated stars of apparent magnitudes 3.3 and 3.5, matching their brightness to Megrez (the dimmest star in the Big Dipper) and Tau Ceti. Participants indicated whether each star was visible from a series of fixation locations, providing a comprehensive map of detection performance in all directions. Contrary to prior predictions, maximum detection was first achieved at ~8° from the star, much closer to the fovea than expected from rod-cone distributions alone. These findings challenge the assumption of optimal detection at the rod density peak and provide the first systematic assessment of an age-old facet of human vision.

An early-morning gene network controlled by phytochromes and cryptochromes regulates photomorphogenesis pathways in Arabidopsis.

Light perception at dawn plays a key role in coordinating multiple molecular processes and in entraining the plant circadian clock. The Arabidopsis mutant lacking the main photoreceptors, however, still shows clock entrainment, indicating that the integration of light into the morning transcriptome is not well understood. In this study, we performed a high-resolution RNA-sequencing time-series experiment, sampling every 2 min beginning at dawn. In parallel experiments, we perturbed temperature, the circadian clock, photoreceptor signaling, and chloroplast-derived light signaling. We used these data to infer a gene network that describes the gene expression dynamics after light stimulus in the morning, and then validated key edges. By sampling time points at high density, we are able to identify three light- and temperature-sensitive bursts of transcription factor activity, one of which lasts for only about 8 min. Phytochrome and cryptochrome mutants cause a delay in the transcriptional bursts at dawn, and completely remove a burst of expression in key photomorphogenesis genes (HY5 and BBX family). Our complete network is available online (http://www-users.york.ac.uk/∼de656/dawnBurst/dawnBurst.html). Taken together, our results show that phytochrome and cryptochrome signaling is required for fine-tuning the dawn transcriptional response to light, but separate pathways can robustly activate much of the program in their absence.

Microsaccades mediate perceptual alternations in Monet's "Impression, sunrise".

Troxler fading, the perceptual disappearance of stationary images upon sustained fixation, is common for objects with equivalent luminance to that of the background. Previous work showed that variations in microsaccadic rates underlie the perceptual vanishing and intensification of simple stimuli, such as Gabor patches. Here, we demonstrate that microsaccade dynamics also contribute to Troxler fading and intensification during the viewing of representational art. Participants fixated a small spot while viewing either a Gabor patch on a blank background, or Monet's painting "Impression, Sunrise." They continuously reported, via button press/release, whether the Gabor patch, or the sun in Monet's painting, was fading versus intensifying, while their eye movements were recorded with high precision. Microsaccade rates peaked before reports of increased visibility, and dropped before reports of decreased visibility or fading, both when viewing Gabor patches and Monet's sun. These results reveal that the relationship between microsaccade production and the reversal and prevention of Troxler fading applies not only to the viewing of contrived stimuli, but also to the observation of "Impression, Sunrise." Whether or not perceptual fading was consciously intended by Monet, our findings indicate that observers' oculomotor dynamics are a contributor to the cornerstone of Impressionism.

What do radiologists look for? Advances and limitations of perceptual learning in radiologic search.

Supported by guidance from training during residency programs, radiologists learn clinically relevant visual features by viewing thousands of medical images. Yet the precise visual features that expert radiologists use in their clinical practice remain unknown. Identifying such features would allow the development of perceptual learning training methods targeted to the optimization of radiology training and the reduction of medical error. Here we review attempts to bridge current gaps in understanding with a focus on computational saliency models that characterize and predict gaze behavior in radiologists. There have been great strides toward the accurate prediction of relevant medical information within images, thereby facilitating the development of novel computer-aided detection and diagnostic tools. In some cases, computational models have achieved equivalent sensitivity to that of radiologists, suggesting that we may be close to identifying the underlying visual representations that radiologists use. However, because the relevant bottom-up features vary across task context and imaging modalities, it will also be necessary to identify relevant top-down factors before perceptual expertise in radiology can be fully understood. Progress along these dimensions will improve the tools available for educating new generations of radiologists, and aid in the detection of medically relevant information, ultimately improving patient health.

A MRI-Based Toolbox for Neurosurgical Planning in Nonhuman Primates.

In this paper, we outline a method for surgical preparation that allows for the practical planning of a variety of neurosurgeries in NHPs solely using data extracted from magnetic resonance imaging (MRI). This protocol allows for the generation of 3D printed anatomically accurate physical models of the brain and skull, as well as an agarose gel model of the brain modeling some of the mechanical properties of the brain. These models can be extracted from MRI using brain extraction software for the model of the brain, and custom code for the model of the skull. The preparation protocol takes advantage of state-of-the-art 3D printing technology to make interfacing brains, skulls, and molds for gel brain models. The skull and brain models can be used to visualize brain tissue inside the skull with the addition of a craniotomy in the custom code, allowing for better preparation for surgeries directly involving the brain. The applications of these methods are designed for surgeries involved in neurological stimulation and recording as well as injection, but the versatility of the system allows for future expansion of the protocol, extraction techniques, and models to a wider scope of surgeries.

Spatiotemporal functional organization of excitatory synaptic inputs onto macaque V1 neurons.

The integration of synaptic inputs onto dendrites provides the basis for neuronal computation. Whereas recent studies have begun to outline the spatial organization of synaptic inputs on individual neurons, the underlying principles related to the specific neural functions are not well understood. Here we perform two-photon dendritic imaging with a genetically-encoded glutamate sensor in awake monkeys, and map the excitatory synaptic inputs on dendrites of individual V1 superficial layer neurons with high spatial and temporal resolution. We find a functional integration and trade-off between orientation-selective and color-selective inputs in basal dendrites of individual V1 neurons. Synaptic inputs on dendrites are spatially clustered by stimulus feature, but functionally scattered in multidimensional feature space, providing a potential substrate of local feature integration on dendritic branches. Furthermore, apical dendrite inputs have larger receptive fields and longer response latencies than basal dendrite inputs, suggesting a dominant role for apical dendrites in integrating feedback in visual information processing.

Microsaccades: Empirical Research and Methodological Advances - Introduction to Part 1 of the Thematic Special Issue.

Recent technical developments and increased affordability of high-speed eye tracking devices have brought microsaccades to the forefront of research in many areas of sensory, perceptual, and cognitive processes. The present thematic issue on "Microsaccades: Empirical Research and Methodological Advances" invited authors to submit original research and reviews encompassing measurements and data analyses in fundamental, translational, and applied studies. We present the first volume of this special issue, comprising 14 articles by research teams around the world. Contributions include the characterization of fixational eye movements and saccadic intrusions in neurological impairments and in visual disease, methodological developments in microsaccade detection, the measurement of fixational eye movements in applied and ecological scenarios, and advances in the current understanding of the relationship between microsaccades and cognition. When fundamental research on microsaccades experienced a renaissance at the turn of the millennium (c.f. 4), one could hardly have been so bold as to predict the manifold applications of research on fixational eye movements in clinic and practice. Through this great variety of areas of focus, some main topics emerge. One such theme is the applicability of microsaccade measures to neurological and visual disease. Whereas microsaccade quantifications have been largely limited to participants with intact visual and oculomotor systems, recent research has extended this interest into the realm of neural and ophthalmic impairment (see 1 for a review). In this volume, Becker et al analyze " Saccadic intrusions in amyotrophic lateral sclerosis (ALS)" and Kang et al study " Fixational eye movement waveforms in amblyopia", delving into the characteristics of fast and slow eye movements. Two other articles focus on how the degradation of visual information, which is relevant to many ophthalmic pathologies, affects microsaccadic features. Tang et al investigate the " Effects of visual blur on microsaccades on visual exploration" and conclude that the precision of an image on the fovea plays an important role in the calibration of microsaccade amplitudes during visual scanning. Otero-Millan et al use different kinds of visual stimuli and viewing tasks in the presence or absence of simulated scotomas, to determine the contributions of foveal and peripheral visual information to microsaccade production. They conclude that " Microsaccade generation requires a foveal anchor". The link between microsaccadic characteristics and cognitive processes has been a mainstay of microsaccade research for almost two decades, since studies in the early 2000s connected microsaccade directions to the spatial location of covert attentional cues (2 3). In the present volume, Dalmaso et al report that " Anticipation of cognitive conflict is reflected in microsaccades", providing new insights about the top-down modulation of microsaccade dynamics. Ryan et al further examine the relationship between " Microsaccades and covert attention" during the performance of a continuous, divided-attention task, and find preliminary evidence that microsaccades track the ongoing allocation of spatial attention. Krueger et al discover that microsaccade rates modulate with visual attention demands and report that " Microsaccades distinguish looking from seeing". Taking the ecological validity of microsaccade investigations one step further, Barnhart et al evaluate microsaccades during the observation of magic tricks and conclude that " Microsaccades reflect the dynamics of misdirected attention in magic". Two articles examine the role of individual differences and intraindividual variability over time on microsaccadic features. In " Reliability and correlates of intra-individual variability in the oculomotor system" Perquin and Bompas find evidence for intra-individual reliability over different time points, while cautioning that its use to classify self-reported individual differences remains unclear. Stafford et al provide a counterpoint in " Can microsaccade rate predict drug response?" by supporting the use of microsaccade occurrence as both a trait measure of individual differences and as a state measure of response to caffeine administration. Methodological and technical advances are the subjects of three papers in this volume. In " Motion tracking of iris features to detect small eye movements" Chaudhary and Pelz describe a new video-based eye tracking methodology that relies on higher-order iris texture features, rather than on lower-order pupil center and corneal reflection features, to detect microsaccades with high confidence. Munz et al present an open source visual analytics system called " VisME: Visual microsaccades explorer" that allows users to interactively vary microsaccade filter parameters and evaluate the resulting effects on microsaccade behavior, with the goal of promoting reproducibility in data analyses. In " What makes a microsaccade? A review of 70 years research prompts a new detection method" Hauperich et al review the microsaccade properties reported between the 1940s and today, and use the stated range of parameters to develop a novel method of microsaccade detection. Lastly, Alexander et al switch the focus from the past of microsaccade research to its future, by discussing the recent and upcoming applications of fixational eye movements to ecologically-valid and real-world scenarios. Their review " Microsaccades in applied environments: real-world applications of fixational eye movement measurements" covers the possibilities and challenges of taking microsaccade measurements out of the lab and into the field. Microsaccades have engaged the interest of scientists from different backgrounds and disciplines for many decades and will certainly continue to do so. One reason for this fascination might be microsaccades' role as a link between basic sensory processes and high-level cognitive phenomena, making them an attractive focus of interdisciplinary research and transdisciplinary applications. Thus, research on microsaccades will not only endure, but keep evolving as the present knowledge base expands. Part 2 of the special issue on microsaccades is already in progress with articles currently under review and will be published in 2021.