Discovery of a body-wide photosensory array that matures in an adult-like animal and mediates eye-brain-independent movement and arousal.

The ability to respond to light has profoundly shaped life. Animals with eyes overwhelmingly rely on their visual circuits for mediating light-induced coordinated movements. Building on previously reported behaviors, we report the discovery of an organized, eye-independent (extraocular), body-wide photosensory framework that allows even a head-removed animal to move like an intact animal. Despite possessing sensitive cerebral eyes and a centralized brain that controls most behaviors, head-removed planarians show acute, coordinated ultraviolet-A (UV-A) aversive phototaxis. We find this eye-brain-independent phototaxis is mediated by two noncanonical rhabdomeric opsins, the first known function for this newly classified opsin-clade. We uncover a unique array of dual-opsin-expressing photoreceptor cells that line the periphery of animal body, are proximal to a body-wide nerve net, and mediate UV-A phototaxis by engaging multiple modes of locomotion. Unlike embryonically developing cerebral eyes that are functional when animals hatch, the body-wide photosensory array matures postembryonically in "adult-like animals." Notably, apart from head-removed phototaxis, the body-wide, extraocular sensory organization also impacts physiology of intact animals. Low-dose UV-A, but not visible light (ocular-stimulus), is able to arouse intact worms that have naturally cycled to an inactive/rest-like state. This wavelength selective, low-light arousal of resting animals is noncanonical-opsin dependent but eye independent. Our discovery of an autonomous, multifunctional, late-maturing, organized body-wide photosensory system establishes a paradigm in sensory biology and evolution of light sensing.

Molecular machines open cell membranes.

Beyond the more common chemical delivery strategies, several physical techniques are used to open the lipid bilayers of cellular membranes. These include using electric and magnetic fields, temperature, ultrasound or light to introduce compounds into cells, to release molecular species from cells or to selectively induce programmed cell death (apoptosis) or uncontrolled cell death (necrosis). More recently, molecular motors and switches that can change their conformation in a controlled manner in response to external stimuli have been used to produce mechanical actions on tissue for biomedical applications. Here we show that molecular machines can drill through cellular bilayers using their molecular-scale actuation, specifically nanomechanical action. Upon physical adsorption of the molecular motors onto lipid bilayers and subsequent activation of the motors using ultraviolet light, holes are drilled in the cell membranes. We designed molecular motors and complementary experimental protocols that use nanomechanical action to induce the diffusion of chemical species out of synthetic vesicles, to enhance the diffusion of traceable molecular machines into and within live cells, to induce necrosis and to introduce chemical species into live cells. We also show that, by using molecular machines that bear short peptide addends, nanomechanical action can selectively target specific cell-surface recognition sites. Beyond the in vitro applications demonstrated here, we expect that molecular machines could also be used in vivo, especially as their design progresses to allow two-photon, near-infrared and radio-frequency activation.

CODEX, a neural network approach to explore signaling dynamics landscapes.

Current studies of cell signaling dynamics that use live cell fluorescent biosensors routinely yield thousands of single-cell, heterogeneous, multi-dimensional trajectories. Typically, the extraction of relevant information from time series data relies on predefined, human-interpretable features. Without a priori knowledge of the system, the predefined features may fail to cover the entire spectrum of dynamics. Here we present CODEX, a data-driven approach based on convolutional neural networks (CNNs) that identifies patterns in time series. It does not require a priori information about the biological system and the insights into the data are built through explanations of the CNNs' predictions. CODEX provides several views of the data: visualization of all the single-cell trajectories in a low-dimensional space, identification of prototypic trajectories, and extraction of distinctive motifs. We demonstrate how CODEX can provide new insights into ERK and Akt signaling in response to various growth factors, and we recapitulate findings in p53 and TGFß-SMAD2 signaling.

Robust neuronal dynamics in premotor cortex during motor planning.

Neural activity maintains representations that bridge past and future events, often over many seconds. Network models can produce persistent and ramping activity, but the positive feedback that is critical for these slow dynamics can cause sensitivity to perturbations. Here we use electrophysiology and optogenetic perturbations in the mouse premotor cortex to probe the robustness of persistent neural representations during motor planning. We show that preparatory activity is remarkably robust to large-scale unilateral silencing: detailed neural dynamics that drive specific future movements were quickly and selectively restored by the network. Selectivity did not recover after bilateral silencing of the premotor cortex. Perturbations to one hemisphere are thus corrected by information from the other hemisphere. Corpus callosum bisections demonstrated that premotor cortex hemispheres can maintain preparatory activity independently. Redundancy across selectively coupled modules, as we observed in the premotor cortex, is a hallmark of robust control systems. Network models incorporating these principles show robustness that is consistent with data.

When uncertain, does human self-motion decision-making fully utilize complete information?

When forced to choose humans often feel uncertain. Investigations of human perceptual decision-making often employ signal detection theory, which assumes that even when uncertain all available information is fully utilized. However, other studies have suggested or assumed that, when uncertain, human subjects guess totally at random, ignoring available information. When uncertain, do humans simply guess totally at random? Or do humans fully utilize complete information? Or does behavior fall between these two extremes yielding "above chance" performance without fully utilizing complete information? While it is often assumed complete information is fully utilized, even when uncertain, to our knowledge this has never been experimentally confirmed. To answer this question, we combined numerical simulations, theoretical analyses, and human studies performed using a self-motion direction-recognition perceptual decision-making task (did I rotate left or right?). Subjects were instructed to make forced-choice binary (left/right) and trinary (left/right/uncertain) decisions when cued following each stimulus. Our results show that humans 1) do not guess at random when uncertain and 2) make binary and trinary decisions equally well. These findings show that humans fully utilize complete information when uncertain for our perceptual decision-making task. This helps unify signal detection theory and other models of forced-choice decision-making which allow for uncertain responses. NEW & NOTEWORTHY Humans make many perceptual decisions every day. But what if we are uncertain? While many studies assume that humans fully utilize complete information, other studies have suggested and/or assumed that when we're uncertain and forced to decide, information is not fully utilized. While humans tend to perform above chance when uncertain, no earlier study has tested whether available information is fully utilized. Our results show that humans make fully informed decisions even when uncertain.

Generalized receptor law governs phototaxis in the phytoplankton Euglena gracilis.

Phototaxis, the process through which motile organisms direct their swimming toward or away from light, is implicated in key ecological phenomena (including algal blooms and diel vertical migration) that shape the distribution, diversity, and productivity of phytoplankton and thus energy transfer to higher trophic levels in aquatic ecosystems. Phototaxis also finds important applications in biofuel reactors and microbiopropellers and is argued to serve as a benchmark for the study of biological invasions in heterogeneous environments owing to the ease of generating stochastic light fields. Despite its ecological and technological relevance, an experimentally tested, general theoretical model of phototaxis seems unavailable to date. Here, we present accurate measurements of the behavior of the alga Euglena gracilis when exposed to controlled light fields. Analysis of E. gracilis' phototactic accumulation dynamics over a broad range of light intensities proves that the classic Keller-Segel mathematical framework for taxis provides an accurate description of both positive and negative phototaxis only when phototactic sensitivity is modeled by a generalized "receptor law," a specific nonlinear response function to light intensity that drives algae toward beneficial light conditions and away from harmful ones. The proposed phototactic model captures the temporal dynamics of both cells' accumulation toward light sources and their dispersion upon light cessation. The model could thus be of use in integrating models of vertical phytoplankton migrations in marine and freshwater ecosystems, and in the design of bioreactors.

Structured-Light-Based System for Shape Measurement of the Human Body in Motion.

The existing methods for measuring the shape of the human body in motion are limited in their practical application owing to immaturity, complexity, and/or high price. Therefore, we propose a method based on structured light supported by multispectral separation to achieve multidirectional and parallel acquisition. Single-frame fringe projection is employed in this method for detailed geometry reconstruction. An extended phase unwrapping method adapted for measurement of the human body is also proposed. This method utilizes local fringe parameter information to identify the optimal unwrapping path for reconstruction. Subsequently, we present a prototype 4DBODY system with a working volume of 2.0 × 1.5 × 1.5 m³, a measurement uncertainty less than 0.5 mm and an average spatial resolution of 1.0 mm for three-dimensional (3D) points. The system consists of eight directional 3D scanners functioning synchronously with an acquisition frequency of 120 Hz. The efficacy of the proposed system is demonstrated by presenting the measurement results obtained for known geometrical objects moving at various speeds as well actual human movements.

Impact of beam angle choice on pencil beam scanning breath-hold proton therapy for lung lesions.

INTRODUCTION: The breath-hold technique inter alia has been suggested to mitigate the detrimental effect of motion on pencil beam scanned (PBS) proton therapy dose distributions. The aim of this study was to evaluate the robustness of incident proton beam angles to day-to-day anatomical variations in breath-hold. MATERIALS AND METHODS: Single field PBS plans at five degrees increments in the transversal plane were made and water-equivalent path lengths (WEPLs) were derived on the planning breath-hold CT (BHCT) for 30 patients diagnosed with locally-advanced non-small cell lung cancer (NSCLC), early stage NSCLC or lung metastasis. Our treatment planning system was subsequently used to recalculate the plans and derive WEPL on a BHCT scan acquired at the end of the treatment. Changes to the V95%, D95 and mean target dose were evaluated. RESULTS: The difference in WEPL as a function of the beam angle was highly patient specific, with a median of 3.3 mm (range: 0.0-41.1 mm). Slightly larger WEPL differences were located around the lateral or lateral anterior/posterior beam angles. Linear models revealed that changes in dose were associated to the changes in WEPL and the tumor baseline shift (p < 0.05). CONCLUSIONS: WEPL changes and tumor baseline shift can serve as reasonable surrogates for dosimetric uncertainty of the target coverage and are well-suited for routine evaluation of plan robustness. The two lateral beam angles are not recommended to use for PBS proton therapy of lung cancer patients treated in breath-hold, due to the poor robustness for several of the patients evaluated.

Intrafractional baseline drift during free breathing breast cancer radiation therapy.

BACKGROUND: Intrafraction motion in breast cancer radiation therapy (BCRT) has not yet been thoroughly described in the literature. It has been observed that baseline drift occurs as part of the intrafraction motion. This study aims to measure baseline drift and its incidence in free-breathing BCRT patients using an in-house developed laser system for tracking the position of the sternum. MATERIALS AND METHODS: Baseline drift was monitored in 20 right-sided breast cancer patients receiving free breathing 3D-conformal RT by using an in-house developed laser system which measures one-dimensional distance in the AP direction. A total of 357 patient respiratory traces from treatment sessions were logged and analysed. Baseline drift was compared to patient positioning error measured from in-field portal imaging. RESULTS: The mean overall baseline drift at end of treatment sessions was -1.3 mm for the patient population. Relatively small baseline drift was observed during the first fraction; however it was clearly detected already at the second fraction. Over 90% of the baseline drift occurs during the first 3 min of each treatment session. The baseline drift rate for the population was -0.5 ± 0.2 mm/min in the posterior direction the first minute after localization. Only 4% of the treatment sessions had a 5 mm or larger baseline drift at 5 min, all towards the posterior direction. Mean baseline drift in the posterior direction in free breathing BCRT was observed in 18 of 20 patients over all treatment sessions. CONCLUSIONS: This study shows that there is a substantial baseline drift in free breathing BCRT patients. No clear baseline drift was observed during the first treatment session; however, baseline drift was markedly present at the rest of the sessions. Intrafraction motion due to baseline drift should be accounted for in margin calculations.

Beam angle evaluation to improve inter-fraction motion robustness in pelvic lymph node irradiation with proton therapy.

BACKGROUND: Proton therapy dose distributions are sensitive to range variations, e.g. arising from inter-fraction organ motion. The aim of this study was to evaluate the inter-fraction motion robustness of proton beams from different beam angles in irradiation of pelvic lymph nodes (LNs). MATERIAL AND METHODS: Planning CT (pCT) and multiple repeat CT (rCT) scans of 18 prostate cancer patients were used. Considering left and right LNs separately, the average water equivalent path length (WEPL) over all ray paths in the beams eye view of the LNs were calculated for all gantry/couch angle combinations across all rCTs versus the corresponding pCT. Single beam proton plans were optimized on the pCT for all gantry angles (0° couch) and were re-calculated on all rCTs for each respective patient. WEPL and dose parameters were extracted and a statistical clustering analysis performed to identify patient sub-populations in terms of patterns in which angles were robust. RESULTS: The WEPL analysis showed a general pattern of least variation for 0° couch beam angles where three minima were found across gantry angles for the left LNs and two for the right LNs. The clustering analysis identified three patient sub-groups for the left LNs and two groups for the right LNs. The dose calculations showed similar results as the WEPL variation, e.g. for the left LNs angles around 25°-35°, 100°-110°, and 160°-170° were consistently preferable for both target and organs at risk. CONCLUSIONS: Sub-populations of patients with similar patterns of WEPL variations across beam angles were identified. The most robust angles found for WEPL variations were also confirmed by the dose/volume analysis.

Effects of intense pulsed light in microstomia in patients with systemic sclerosis: A pilot study.

INTRODUCTION: The effects of intense pulsed light (IPL) on collagen structures are well known in the treatment of photodamaged skin. OBJECTIVE: The objective of this study was to investigate the effect of IPL on sclerotic skin by treating patients with microstomia due to systemic sclerosis. METHODS AND MATERIALS: 13 patients all with microstomia and systemic sclerosis were treated with IPL, PR (530-750 nm filter) and/or VL (555-950 nm filter) applicator. They were treated in the perioral area 8 times with 3-4 weeks of interval and follow-up for 6 months. The outcomes were the inter-incisal distance and the inter-ridge distance. RESULTS: A significant increase in mouth opening of 4.1 mm (95% confidence interval, 1726-6638, p < 0.005) was found in the inter-ridge distance when comparing the distance before treatment with the distance at six-month follow-up. No significant difference was found in the inter-incisal distance. The patients experienced improved mobility and better control of lip movements after the treatments. CONCLUSION: IPL can improve the inter-ridge distance between the lips in patients with microstomia due to systemic sclerosis but does not affect the inter-incisal distance, which is also dependent on the mobility of the mandibular joints. This treatment can be considered an adjunctive therapy in patients with microstomia due to systemic sclerosis.

Actin-dependent plastid movement is required for motive force generation in directional nuclear movement in plants.

Nuclear movement and positioning are indispensable for most cellular functions. In plants, strong light-induced chloroplast movement to the side walls of the cell is essential for minimizing damage from strong visible light. Strong light-induced nuclear movement to the side walls also has been suggested to play an important role in minimizing damage from strong UV light. Although both movements are regulated by the same photoreceptor, phototropin, the precise cytoskeleton-based force generation mechanism for nuclear movement is unknown, in contrast to the short actin-based mechanism of chloroplast movement. Here we show that actin-dependent movement of plastids attached to the nucleus is essential for light-induced nuclear movement in the Arabidopsis leaf epidermal cell. We found that nuclei are always associated with some plastids, and that light-induced nuclear movement is correlated with the dynamics of short actin filaments associated with plastids. Indeed, nuclei without plastid attachments do not exhibit blue light-induced directional movement. Our results demonstrate that nuclei are incapable of autonomously moving in response to light, whereas attached plastids carry nuclei via the short actin filament-based movement. Thus, the close association between nuclei and plastids is essential for their cooperative movements and functions.

Sensorimotor structure of Drosophila larva phototaxis.

The avoidance of light by fly larvae is a classic paradigm for sensorimotor behavior. Here, we use behavioral assays and video microscopy to quantify the sensorimotor structure of phototaxis using the Drosophila larva. Larval locomotion is composed of sequences of runs (periods of forward movement) that are interrupted by abrupt turns, during which the larva pauses and sweeps its head back and forth, probing local light information to determine the direction of the successive run. All phototactic responses are mediated by the same set of sensorimotor transformations that require temporal processing of sensory inputs. Through functional imaging and genetic inactivation of specific neurons downstream of the sensory periphery, we have begun to map these sensorimotor circuits into the larval central brain. We find that specific sensorimotor pathways that govern distinct light-evoked responses begin to segregate at the first relay after the photosensory neurons.

Chloroplast and nuclear photorelocation movements.

Chloroplasts move toward weak light to increase photosynthetic efficiency, and migrate away from strong light to protect chloroplasts from photodamage and eventual cell death. These chloroplast behaviors were first observed more than 100 years ago, but the underlying mechanism has only recently been identified. Ideal plant materials, such as fern gametophytes for photobiological and cell biological approaches, and Arabidopsis thaliana for genetic analyses, have been used along with sophisticated methods, such as partial cell irradiation and time-lapse video recording under infrared light to study chloroplast movement. These studies have revealed precise chloroplast behavior, and identified photoreceptors, other relevant protein components, and novel actin filament structures required for chloroplast movement. In this review, our findings regarding chloroplast and nuclear movements are described.

A Simulation Study of a Radiofrequency Localization System for Tracking Patient Motion in Radiotherapy.

One of the most widely used tools in cancer treatment is external beam radiotherapy. However, the major risk involved in radiotherapy is excess radiation dose to healthy tissue, exacerbated by patient motion. Here, we present a simulation study of a potential radiofrequency (RF) localization system designed to track intrafraction motion (target motion during the radiation treatment). This system includes skin-wearable RF beacons and an external tracking system. We develop an analytical model for direction of arrival measurement with radio frequencies (GHz range) for use in a localization estimate. We use a Monte Carlo simulation to investigate the relationship between a localization estimate and angular resolution of sensors (signal receivers) in a simulated room. The results indicate that the external sensor needs an angular resolution of about 0.03 degrees to achieve millimeter-level localization accuracy in a treatment room. This fundamental study of a novel RF localization system offers the groundwork to design a radiotherapy-compatible patient positioning system for active motion compensation.

Common behaviors alterations after extremely low-frequency electromagnetic field exposure in rat animal model.

Naturally, the presence of electromagnetic waves in our living environment affects all components of organisms, particularly humans and animals, as the large part of their body consists of water. In the present study, we tried to investigate the relation between exposure to the extremely low-frequency electromagnetic field (ELF-EMF) and common behaviors such as body weight, food and water intake, anorexia (poor appetite), plasma glucose concentration, movement, rearing and sniffing in rats. For this purpose, rats were exposed to 40 Hz ELF-EMF once a day for 21 days, then at days 1, 3, 7, 14 and 21 after exposure, any changes in the above-mentioned items were assessed in the exposed rats and compared to the non-exposed group as control. Body weight of irradiated rats significantly increased only a week after exposure and decreased after that. No significant change was observed in food and water intake of irradiated rats compared to the control, and the anorexia parameter in the group exposed to ELF-EMF was significantly decreased at one and two weeks after irradiation. A week after exposure, the level of glucose was significantly increased but at other days these changes were not significant. Movements, rearing and sniffing of rats at day 1 after exposure were significantly decreased and other days these changes did not follow any particular pattern. However, the result of this study demonstrated that exposure to ELF-EMF can alter the normal condition of animals and may represent a harmful impact on behavior.

Behaviour of the plathelminth Symsagittifera roscoffensis under different light conditions and the consequences for the symbiotic algae Tetraselmis convolutae.

Symsagittifera roscoffensis is a plathelminth living in symbiosis with the green algae Tetraselmis convolutae. Host and symbiont are a model system for the study of endosymbiosis, which has so far mainly focused on their biochemical interactions. Symsagittifera roscoffensis is well known for its positive phototaxis that is hypothesized to optimize the symbiont's light perception for photosynthesis. In this study, we conducted a detailed analysis of phototaxis using light sources of different wavelength and brightness by videotracking. Furthermore, we compared the behavioural data with the electron transfer rate of the photosystem from cultured symbiotic cells. The symbiotic algae is adapted to low light conditions, showing a positive electron transfer rate at a photosynthetically active radiation of 0.112 µmol photons m(-2) s(-1), and S. roscoffensis showed a positive phototactic behaviour for light intensities up to 459.17 µmol photons m(-2) s(-1), which is not optimal regarding the needs of the symbiotic cells and may even harm host and symbiont. Red light cannot be detected by the animals and therefore their eyes seem not to be suitable for measuring the exact photosynthetically active radiation to the benefit of the photosymbionts.

An Optimized Biological Taser: Electric Eels Remotely Induce or Arrest Movement in Nearby Prey.

Despite centuries of interest in electric eels, few studies have investigated the mechanism of the eel's attack. Here, I review and extend recent findings that show eel electric high-voltage discharges activate prey motor neuron efferents. This mechanism allows electric eels to remotely control their targets using two different strategies. When nearby prey have been detected, eels emit a high-voltage volley that causes whole-body tetanus in the target, freezing all voluntary movement and allowing the eel to capture the prey with a suction feeding strike. When hunting for cryptic prey, eels emit doublets and triplets, inducing whole-body twitch in prey, which in turn elicits an immediate eel attack with a full volley and suction feeding strike. Thus, by using their modified muscles (electrocytes) as amplifiers of their own motor efferents, eel's motor neurons remotely activate prey motor neurons to cause movement (twitch and escape) or immobilization (tetanus) facilitating prey detection and capture, respectively. These results explain reports that human movement is 'frozen' by eel discharges and shows the mechanism to resemble a law-enforcement Taser.

Clues to the signals for chloroplast photo-relocation from the lifetimes of accumulation and avoidance responses.

Chloroplast photo-relocation movement is crucial for plant survival; however, the mechanism of this phenomenon is still poorly understood. Especially, the signal that goes from photoreceptor to chloroplast is unknown, although the photoreceptors (phototropin 1 and 2) have been identified and an actin structure (chloroplast actin filaments) has been characterized that is specific for chloroplast movement. Here, in gametophytes of the fern Adiantum capillus-veneris, gametophores of the moss Physcomiterella patens, and leaves of the seed plant Arabidopsis thaliana, we sought to characterize the signaling system by measuring the lifetime of the induced response. Chloroplast movements were induced by microbeam irradiation with high-intensity blue light and recorded. The lifetime of the avoidance state was measured as a lag time between switching off the beam and the loss of avoidance behavior, and that of the accumulation state was measured as the duration of accumulation behavior following the extinction of the beam. The lifetime for the avoidance response state is approximately 3-4 min and that for the accumulation response is 19-28 min. These data suggest that the two responses are based on distinct signals.

Identification of nonvisual photomotor response cells in the vertebrate hindbrain.

Nonvisual photosensation enables animals to sense light without sight. However, the cellular and molecular mechanisms of nonvisual photobehaviors are poorly understood, especially in vertebrate animals. Here, we describe the photomotor response (PMR), a robust and reproducible series of motor behaviors in zebrafish that is elicited by visual wavelengths of light but does not require the eyes, pineal gland, or other canonical deep-brain photoreceptive organs. Unlike the relatively slow effects of canonical nonvisual pathways, motor circuits are strongly and quickly (seconds) recruited during the PMR behavior. We find that the hindbrain is both necessary and sufficient to drive these behaviors. Using in vivo calcium imaging, we identify a discrete set of neurons within the hindbrain whose responses to light mirror the PMR behavior. Pharmacological inhibition of the visual cycle blocks PMR behaviors, suggesting that opsin-based photoreceptors control this behavior. These data represent the first known light-sensing circuit in the vertebrate hindbrain.