Onset Of Signal Research Articles

Stage-specific CAR-mediated signaling generates naïve-like, TCR-null CAR T cells from induced pluripotent stem cells.

Genetically modified, induced pluripotent stem cells (iPSCs) offer a promising allogeneic source for the generation of functionally enhanced, chimeric antigen receptor (CAR) T cells. However, the signaling of CARs during early T cell development and the removal of the endogenous T cell receptor required to prevent alloreactivity pose significant challenges to the production of mature conventional CAR T cells from iPSCs. Here, we show that TCR-null, CD8αβ CAR T cells can be efficiently generated from iPSCs by engineering stage-specific onset of CAR expression and signaling to both permit conventional T cell development and to induce efficient positive selection. CAR T cells produced using this approach displayed a uniform, naïve T cell phenotype and demonstrated superior antigen-specific cytotoxicity compared to iPSC-derived effector memory CAR T cells. Multimodal sequencing revealed CAR-mediated positive selection induced the persistent upregulation of key transcription factors involved in naïve T cell development. Achieving precise control of CAR expression and signaling in developmentally sensitive T precursors will be critical to realizing the full potential for "off-the-shelf", iPSC-derived cellular therapies.

Rapid Onset of Innate Response, Cytokine Signaling and Humoral Immunity in Inactivated LPAI-H9N2-Vaccinated Broilers

The development of effective and innovative vaccination strategies is urgently needed to better control the spread and transmission of the low-pathogenic avian influenza H9N2 subtype (LPAI-H9N2) in poultry. In addition, the enhancement of innate immunity by some of these innovative inactivated vaccines has not yet been investigated. Here, an experiment was conducted in commercial broiler chickens to compare the immune response to two different inactivated H9N2 vaccines. For this, Group 1 (G1) broilers were vaccinated with vaccine 1 [Nobilis® H9N2-P (pathogen-associated molecular patterns—PAMP) technology], broilers in G2 were vaccinated with vaccine 2 [an inactivated whole H9N2 virus (IWV) autogenous oil emulsion vaccine], while birds in G3 were not vaccinated. The study lasted 34 days. Innate immune parameters (phagocytic activity, nitric oxide, and lysozyme), cytokine signaling (IL-1β, IL-6, IL-8), humoral immunity using the hemagglutination inhibition (HI) test, and the gene expressions of IFN-γ and TLR-21 were assessed. The results showed a significant increase in innate immunity and modulatory cytokines at 24–48 h after the vaccination of G1 broilers, with a continuous increase until the end of the experiment. In addition, a significant increase in geometric mean HI titers was observed in G1 at 11 days post-vaccination (dpv), and a significant (p ˂ 0.05) upregulation of IFN-γ and TLR-21 was observed in the same group, G1, at 31 dpv compared to G2 and G3. Nobilis® H9N2-P may induce faster and stronger innate and active humoral immunity compared to another IWV, which may contribute to the protection of broilers against early H9N2 infections. However, challenge protection studies for several IWV vaccines, including PAMP-H9N2 against LPAI-H9N2, should be further evaluated in both specific pathogen-free (SPF) and commercial broilers.

Initiation and maintenance of the pluripotent epiblast in pre-implantation human development is independent of NODAL signaling.

The human blastocyst contains the pluripotent epiblast from which human embryonic stem cells (hESCs) can be derived. ACTIVIN/NODAL signaling maintains expression of the transcription factor NANOG and invitro propagation of hESCs. It is unknown whether this reflects a functional requirement for epiblast development in human embryos. Here, we characterized NODAL signaling activity during pre-implantation human development. We showed that NANOG is an early molecular marker restricted to the nascent human pluripotent epiblast and was initiated prior to the onset of NODAL signaling. We further demonstrated that expression of pluripotency-associated transcription factors NANOG, SOX2, OCT4, and KLF17 were maintained in the epiblast in the absence of NODAL signaling activity. Genome-wide transcriptional analysis showed that NODAL signaling inhibition did not decrease NANOG transcription or impact the wider pluripotency-associated gene regulatory network. These data suggest differences in the signaling requirements regulating pluripotency in the pre-implantation human epiblast compared with existing hESC culture.

Impairment of neuronal tyrosine phosphatase STEP worsens post-ischemic inflammation and brain injury under hypertensive condition

Hypertension is associated with poor outcome and higher mortality in patients with ischemic stroke. The impairment of adaptive vascular mechanisms under hypertensive condition compromises collateral blood flow after arterial occlusion in patients with acute ischemic stroke resulting in hypoperfusion. The increased oxidative stress caused by hypoperfusion is thought to be a trigger for the rapid evolution of ischemic infarct volume under hypertensive condition. However, the cellular factors and pathways that contribute to the exacerbation of ischemic brain injury under hypertensive condition is not yet understood. The current study reveals that predisposition to hypertension leads to basal loss of function of the neuron-specific tyrosine phosphatase STEP, which plays a crucial role in neuroprotection against excitotoxic insult. The findings further show that a mild ischemic insult in hypertensive rats triggers an early onset and sustained activation of the neuronal extracellular signal regulated kinase (ERK MAPK), a member of the mitogen activated protein kinase family and a substrate of STEP. This leads to rapid increase in the activation of neuronal NF-κB, expression of neuronal cyclooxygenase-2 and subsequent biosynthesis of the pro-inflammatory mediator prostaglandin E2, resulting in rapid morphological transformation of microglia to the pro-inflammatory state and subsequent exacerbation of ischemic brain injury. Restoration of STEP signaling with intravenous administration of a STEP-derived peptide mimetic reduces the pro-inflammatory response in neurons, activation of microglia, and ischemic brain injury. The findings suggest that the basal loss of STEP function under hypertensive condition contributes to the exacerbation of ischemic brain injury by enhancing post-ischemic inflammatory response. The study not only presents a novel role of STEP in regulating neuroimmune communication but also highlights the therapeutic potential of a STEP-mimetic in mitigating ischemic brain damage under hypertensive condition.

Enhancing Safety in Autonomous Vehicles: The Impact of Auditory and Visual Warning Signals on Driver Behavior and Situational Awareness

Semi-autonomous vehicles (AVs) enable drivers to engage in non-driving tasks but require them to be ready to take control during critical situations. This “out-of-the-loop” problem demands a quick transition to active information processing, raising safety concerns and anxiety. Multimodal signals in AVs aim to deliver take-over requests and facilitate driver–vehicle cooperation. However, the effectiveness of auditory, visual, or combined signals in improving situational awareness and reaction time for safe maneuvering remains unclear. This study investigates how signal modalities affect drivers’ behavior using virtual reality (VR). We measured drivers’ reaction times from signal onset to take-over response and gaze dwell time for situational awareness across twelve critical events. Furthermore, we assessed self-reported anxiety and trust levels using the Autonomous Vehicle Acceptance Model questionnaire. The results showed that visual signals significantly reduced reaction times, whereas auditory signals did not. Additionally, any warning signal, together with seeing driving hazards, increased successful maneuvering. The analysis of gaze dwell time on driving hazards revealed that audio and visual signals improved situational awareness. Lastly, warning signals reduced anxiety and increased trust. These results highlight the distinct effectiveness of signal modalities in improving driver reaction times, situational awareness, and perceived safety, mitigating the “out-of-the-loop” problem and fostering human–vehicle cooperation.

What would affect drivers’ stop-and-go decisions at yellow dilemma zones? A driving simulator study in Hong Kong

Yellow dilemma, at which a driver can neither stop nor go safely after the onset of yellow signals, is one of the major crash contributory factors at the signal junctions. Studies have visited the yellow dilemma problem using observation surveys. Factors including road environment, traffic conditions, and driver characteristics that affect the driver behaviours are revealed. However, it is rare that the joint effects of situational and attitudinal factors on the driver behaviours at the yellow dilemma zone are considered. In this study, drivers’ propensity to stop after the onset of yellow signals is examined using the driving simulator approach. For instances, the association between driver propensity, socio-demographics, safety perception, traffic signals, and traffic and weather conditions are measured using a binary logit model. Additionally, variations in the effect of influencing factors on driver behaviours are accommodated by adding the interaction terms for driver characteristics, traffic flow characteristics, traffic signals, and weather conditions. Results indicate that weather conditions, traffic volume, position of yellow dilemma in the sequence, driver age and safety perception significantly affect the drivers’ propensity to stop after the onset of yellow signals. Furthermore, there are remarkable interactions for the effects of driver gender and location of yellow dilemma.

Exploring the role of spinal astrocytes in the onset of hyperalgesic priming signals in acid-induced chronic muscle pain.

Hyperalgesic priming, a form of pain plasticity initiated by initial injury, leads to heightened sensitivity to subsequent noxious stimuli, contributing to chronic pain development in animals. While astrocytes play active roles in modulating synaptic transmission in various pain models, their specific involvement in hyperalgesic priming remains elusive. Here, we show that spinal astrocytes are essential for hyperalgesic priming formation in a mouse model of acid-induced muscle pain. We observed spinal astrocyte activation 4 h after initial acid injection, and inhibition of this activation prevented chronic pain development upon subsequent acid injection. Chemogenetic activation of spinal astrocytes mimicked the first acid-induced hyperalgesic priming. We also demonstrated that spinal phosphorylated extracellular regulated kinase (pERK)-positive neurons were mainly vesicular glutamate transporter-2 positive (Vglut2+) neurons after the first acid injection, and inhibition of spinal pERK prevented astrocyte activation. Furthermore, pharmacological inhibition of astrocytic glutamate transporters glutamate transporter-1 and glutamate-aspartate transporter abolished the hyperalgesic priming. Collectively, our results suggest that pERK activation in Vglut2+ neurons activate astrocytes through astrocytic glutamate transporters. This process eventually establishes hyperalgesic priming through spinal D-serine. We conclude that spinal astrocytes play a crucial role in the transition from acute to chronic pain.

Iterative alignment discovery of speech-associated neural activity.

Objective. Brain-computer interfaces (BCIs) have the potential to preserve or restore speech in patients with neurological disorders that weaken the muscles involved in speech production. However, successful training of low-latency speech synthesis and recognition models requires alignment of neural activity with intended phonetic or acoustic output with high temporal precision. This is particularly challenging in patients who cannot produce audible speech, as ground truth with which to pinpoint neural activity synchronized with speech is not available.Approach. In this study, we present a new iterative algorithm for neural voice activity detection (nVAD) called iterative alignment discovery dynamic time warping (IAD-DTW) that integrates DTW into the loss function of a deep neural network (DNN). The algorithm is designed to discover the alignment between a patient's electrocorticographic (ECoG) neural responses and their attempts to speak during collection of data for training BCI decoders for speech synthesis and recognition.Main results. To demonstrate the effectiveness of the algorithm, we tested its accuracy in predicting the onset and duration of acoustic signals produced by able-bodied patients with intact speech undergoing short-term diagnostic ECoG recordings for epilepsy surgery. We simulated a lack of ground truth by randomly perturbing the temporal correspondence between neural activity and an initial single estimate for all speech onsets and durations. We examined the model's ability to overcome these perturbations to estimate ground truth. IAD-DTW showed no notable degradation (<1% absolute decrease in accuracy) in performance in these simulations, even in the case of maximal misalignments between speech and silence.Significance. IAD-DTW is computationally inexpensive and can be easily integrated into existing DNN-based nVAD approaches, as it pertains only to the final loss computation. This approach makes it possible to train speech BCI algorithms using ECoG data from patients who are unable to produce audible speech, including those with Locked-In Syndrome.

Do microsaccades vary with discriminability around the visual field?

Microsaccades-tiny fixational eye movements-improve discriminability in high-acuity tasks in the foveola. To investigate whether they help compensate for low discriminability at the perifovea, we examined microsaccade characteristics relative to the adult visual performance field, which is characterized by two perceptual asymmetries: horizontal-vertical anisotropy (better discrimination along the horizontal than vertical meridian) and vertical meridian asymmetry (better discrimination along the lower than upper vertical meridian). We investigated whether and to what extent microsaccade directionality varies when stimuli are at isoeccentric locations along the cardinals under conditions of heterogeneous discriminability (Experiment 1) and homogeneous discriminability, equated by adjusting stimulus contrast (Experiment 2). Participants performed a two-alternative forced-choice orientation discrimination task. In both experiments, performance was better on trials without microsaccades between ready signal onset and stimulus offset than on trials with microsaccades. Across the trial sequence, the microsaccade rate and directional pattern were similar across locations. Our results indicate that microsaccades were similar regardless of stimulus discriminability and target location, except during the response period-once the stimuli were no longer present and target location no longer uncertain-when microsaccades were biased toward the target location. Thus, this study reveals that microsaccades do not flexibly adapt as a function of varying discriminability in a basic visual task around the visual field.

Advances in the prerequisite and consequence of STING downstream signalosomes.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway is an evolving DNA-sensing mechanism involved in innate immunity and pathogen defense that has been optimized while remaining conserved. Aside from recognizing pathogens through conserved motifs, these receptors also detect aberrant or misplaced self-molecules as possible signs of perturbed homeostasis. Upon binding external or self-derived DNA, a mobile secondary messenger 2'3'-cyclic GMP-AMP (cGAMP) is produced by cGAS and in turn activates its adapter STING in the endoplasmic reticulum (ER). Resting-state or activated STING protein is finely restricted by multiple degradation machineries. The post-translational changes of the STING protein, along with the regulatory machinery of the secret routes, limit the onset, strength and sustention of STING signal. STING experiences a conformational shift and relocates with TBK1 from the ER to perinuclear vesicles containing transcription factors, provoking the transcription activity of IRF3/IFN-I and NF-κB pathways, as well as to initiate a number of cellular processes that have been shown to alter the immune landscape in cancer, such as autophagy, NLRP3 inflammasome, ER stress, and cell death. STING signal thus serves as a potent activator for immune mobilization yetalso triggers immune-mediated pathology in tissues. Recent advances have established the vital role of STING in immune surveillance as well as tumorigenic process. This review provides an overview of the disparate outcomes of cancer attributed to the actions of pleiotropic and coordinated STING downstream signalosomes, along with the underlying mechanisms of STING function in pathologies, providing therapeutic implications for new approaches in hunt for the next generation of cancer immunotherapy base on STING.

Acoustic emission onset time detection for structural monitoring with U-Net neural network architecture

Acoustic Emission (AE) is a non-destructive structural health monitoring technique, which studies elastic waves emitted during crack formation. Utilizing piezoelectric sensors, these waves are converted into electrical signals for subsequent analysis, offering insights into crack propagation and structural durability. This study focuses on the identification of AE signal onset times, crucial for determining crack locations. Conventional methods often encounter challenges with background noise, prompting the need for innovative approaches. Leveraging a U-Net neural network, specialized in segmentation tasks, onset time identification is approached as a one-dimensional segmentation challenge. Through training and testing on Pencil Lead Break (PLB) test data, commonly used in AE evaluations, the effectiveness of the method is demonstrated even with continuous signals, suggesting potential applicability in real-time monitoring.

Audiovisual speech asynchrony asymmetrically modulates neural binding

Previous psychophysical and neurophysiological studies in young healthy adults have provided evidence that audiovisual speech integration occurs with a large degree of temporal tolerance around true simultaneity. To further determine whether audiovisual speech asynchrony modulates auditory cortical processing and neural binding in young healthy adults, N1/P2 auditory evoked responses were compared using an additive model during a syllable categorization task, without or with an audiovisual asynchrony ranging from 240 ms visual lead to 240 ms auditory lead. Consistent with previous psychophysical findings, the observed results converge in favor of an asymmetric temporal integration window. Three main findings were observed: 1) predictive temporal and phonetic cues from pre-phonatory visual movements before the acoustic onset appeared essential for neural binding to occur, 2) audiovisual synchrony, with visual pre-phonatory movements predictive of the onset of the acoustic signal, was a prerequisite for N1 latency facilitation, and 3) P2 amplitude suppression and latency facilitation occurred even when visual pre-phonatory movements were not predictive of the acoustic onset but the syllable to come. Taken together, these findings help further clarify how audiovisual speech integration partly operates through two stages of visually-based temporal and phonetic predictions.

Index analysis: An approach to understand signal transduction with application to the EGFR signalling pathway.

In systems biology and pharmacology, large-scale kinetic models are used to study the dynamic response of a system to a specific input or stimulus. While in many applications, a deeper understanding of the input-response behaviour is highly desirable, it is often hindered by the large number of molecular species and the complexity of the interactions. An approach that identifies key molecular species for a given input-response relationship and characterises dynamic properties of states is therefore highly desirable. We introduce the concept of index analysis; it is based on different time- and state-dependent quantities (indices) to identify important dynamic characteristics of molecular species. All indices are defined for a specific pair of input and response variables as well as for a specific magnitude of the input. In application to a large-scale kinetic model of the EGFR signalling cascade, we identified different phases of signal transduction, the peculiar role of Phosphatase3 during signal activation and Ras recycling during signal onset. In addition, we discuss the challenges and pitfalls of interpreting the relevance of molecular species based on knock-out simulation studies, and provide an alternative view on conflicting results on the importance of parallel EGFR downstream pathways. Beyond the applications in model interpretation, index analysis is envisioned to be a valuable tool in model reduction.

Differential Interferon Signaling Regulation and Oxidative Stress Responses in the Cerebral Cortex and Cerebellum Could Account for the Spatiotemporal Pattern of Neurodegeneration in Niemann-Pick Disease Type C.

Niemann-Pick disease type C (NPC) is a fatal neurodegenerative condition caused by genetic mutations of the NPC1 or NPC2 genes that encode the NPC1 and NPC2 proteins, respectively, which are believed to be responsible for cholesterol efflux from late-endosomes/lysosomes. The pathogenic mechanisms that lead to neurodegeneration in NPC are not well understood. There are, however, well-defined spatiotemporal patterns of neurodegeneration that may provide insight into the pathogenic process. For example, the cerebellum is severely affected from early disease stages, compared with cerebral regions, which remain relatively spared until later stages. Using a genome-wide transcriptome analysis, we have recently identified an aberrant pattern of interferon activation in the cerebella of pre-symptomatic Npc1-/- mice. Here, we carried out a comparative transcriptomic analysis of cerebral cortices and cerebella of pre-symptomatic Npc1-/- mice and age-matched controls to identify differences that may help explain the pathological progression within the NPC brain. We report lower cerebral expression of genes within interferon signaling pathways, and significant differences in the regulation of oxidative stress, compared with the cerebellum. Our findings suggest that a delayed onset of interferon signaling, possibly linked to lower oxidative stress, may account for the slower onset of cerebral cortical pathology in the disease.

Sound-producing actions in guitar performance of groove-based microrhythm

This paper reports on an experiment that investigated how guitarists signal the intended timing of a rhythmic event in a groove-based context via three different features related to sound-producing motions of impulsive chord strokes (striking velocity, movement duration and fretboard position). 21 expert electric guitarists were instructed to perform a simple rhythmic pattern in three different timing styles—"laidback," "on-the-beat," and "pushed"—in tandem with a metronome. Results revealed systematic differences across participants in the striking velocity and movement duration of chords in the different timing styles. In general, laid-back strokes were played with lower striking velocity and longer movement duration relative to on-the-beat and pushed strokes. No differences in the fretboard striking position were found (either closer to the "bridge" [bottom] or to the "neck" [head]). Correlations with previously-reported audio features of the guitar strokes were also investigated, where lower velocity and longer movement duration generally corresponded with longer acoustic attack duration (signal onset to offset).

Yielding Behavior of Turning Drivers to a Pedestrian With a Cane at Low-Light Signalized Intersections

Introduction: This study evaluated drivers’ delays and yields when making right turns at the onset of a circular green traffic signal, in response to pedestrian behaviors, during daylight and at night with streetlamp lighting, in urban and suburban locations. Methods: Researchers conducted trials at three intersections, two typical urban and one typical suburban intersection. Driver movement data were collected (N = 302) in three conditions: no pedestrian present (control); a pedestrian displaying a cane; and a pedestrian flagging a cane. Driver delay was recorded in seconds from the onset of the green traffic signal to when the vehicle crossed the middle of the width of the pedestrian's crosswalk. Results: The primary result was that there are no statistical differences between day and night driver delay. Secondarily, statistically significant differences were found for urban versus suburban drivers when flagging the cane versus displaying the cane, with urban sites and flagging a cane garnering greater delays. Yield calculations, based on the time required for a pedestrian to nearly clear the first half of the crosswalk, confirmed the patterns of drivers’ behaviors. Discussion: Authors anticipated negative effects on yielding in nighttime lower-light environments; however, there was similar driver delay and yields compared to daylight hours. Greater driver delay and yields, for flagging a cane and at urban settings, confirmed the results in previous studies. Implications for Practitioners: When assessing travel options, informed pedestrians and orientation and mobility specialists can consider the effectiveness of active cane techniques to increase the likelihood of yields from turning drivers, even at night where streetlamp lighting is the source of illumination. When instructing cane techniques for use at crosswalks, specialists can consider the options for mitigating risk when crossing typical streets in urban and suburban areas.

Improving onset picking in ultrasonic testing by using a spectral entropy criterion.

In ultrasonic testing, material and structural properties of a specimen can be derived from the time-of-flight (ToF). Using signal features, such as the first peak or envelope maximum, to calculate the ToF is error-prone in multipath arrangements or dispersive and attenuating materials, which is not the case for the signal onset. Borrowing from seismology, researchers used the Akaike information criterion (AIC) picker to automatically determine onset times. The most commonly used formulation, Maeda's AIC picker, is reassessed and found to be based on inappropriate assumptions for signals often used in ultrasonic testing and dependent on arbitrary parameters. Consequently, an onset picker for ultrasonic through-transmission measurements is proposed, based on a spectral entropy criterion (SEC) to model the signal using the AIC framework. This SEC picker takes into account the spectral properties of the ultrasonic signal and is virtually free of arbitrary parameters. Synthetic and experimental data are used to compare the performance of SEC and AIC pickers. It is shown that the accuracy of onset picking is improved for densely sampled data.

The Influence of the Masker on the Localization of the Moving Signal in the Horizontal Plane

The effect of the masker on the localization of the moving signal was investigated in the free field conditions. The experiments were carried out in an anechoic chamber. Sound signals were presented from loudspeakers located on a semicircular arc in the horizontal plane. Bandpass noise bursts (5–18 kHz) were used to create a signal and a masker. The signal and the mask were uncorrelated stimuli and were created from two independent noise bursts. The stationary masker was always on the right at an angle of 15°. The moving signals traveled to or from the masker along two paths located at two places (–86° to –52° and –52° to –18°). The signal and the masker of 1-s duration each were presented either simultaneously or with a delay of the signal onset relative to the masker onset. The delay varied from 1 to 40 ms and 1200 ms. The subjects evaluated the start and end points of the trajectory of the moving sounds. Localization data for a moving signal under masking conditions were compared with spatial estimates of the same signal when presented in isolation (without a masker). Localization of the start and end points of the signal in masking condition was compared with localization of the moving source alone. Results showed that the masker affected the start and end points of the signal trajectory. The shift depended on the direction of movement. The starting points were always shifted in the direction of motion of the signal. The end points were shifted in the opposite direction.

New proglacial meteorology and river stage observations from Inglefield Land and Pituffik, NW Greenland

Abstract. Meltwater runoff from the Greenland ice sheet (GrIS) is an important contributor to global sea level rise, but substantial uncertainty exists in its measurement and prediction. Common approaches for estimating ice sheet runoff are in situ gauging of proglacial rivers draining the ice sheet and surface mass balance (SMB) modeling. To obtain hydrological and meteorological data sets suitable for both runoff stage characterization and, pending the establishment of stage–discharge curves, SMB model evaluation, we established an automated weather station (AWS) and a cluster of traditional and experimental river stage sensors on the Minturn River, the largest proglacial river draining Inglefield Land, NW Greenland. Secondary installations measuring river stage were installed in the Fox Canyon River and North River at Pituffik Space Base, NW Greenland. Proglacial runoff at these sites is dominated by supraglacial processes only, uniquely advantaging them for SMB studies. The three installations provide rare hydrological time series and an opportunity to evaluate experimental measurements of river stage from a harsh, little-studied polar region. The installed instruments include submerged vented and non-vented pressure transducers, a bubbler sensor, experimental bank-mounted laser rangefinders, and time-lapse cameras. The first 3 years of observations (2019 to 2021) from these stations indicate (a) a meltwater runoff season from late June to late August/early September that is roughly synchronous throughout the region; (b) the early onset (∼ 23 June to 8 July) of a strong diurnal runoff signal in 2019 and 2020, suggesting minimal meltwater storage in snow and/or firn; (c) 1 d lagged air temperature that displays the strongest correlation with river stage; (d) river stage that correlates more strongly with ablation zone albedo than with net radiation; and (e) the late-summer rain-on-ice events appear to trigger the region's sharpest and largest floods. The new gauging stations provide valuable in situ hydrological observations that are freely available through the PROMICE network (https://promice.org/weather-stations/, last access: 14 September 2023).

Nociception in Chicken Embryos, Part II: Embryonal Development of Electroencephalic Neuronal Activity In Ovo as a Prerequisite for Nociception.

Chicken culling has been forbidden in Germany since 2022; male/female selection and male elimination must be brought to an embryonic status prior to the onset of nociception. The present study evaluated the ontogenetic point at which noxious stimuli could potentially be perceived/processed in the brain in ovo. EEG recordings from randomized hyperpallial brain sites were recorded in ovo and noxious stimuli were applied. Temporal and spectral analyses of the EEG were performed. The onset of physiological neuronal signals could be determined at developmental day 13. ERP/ERSP/ITC analysis did not reveal phase-locked nociceptive responses. Although no central nociceptive responses were documented, adequate EEG responses to noxious stimuli from other brain areas cannot be excluded. The extreme stress impact on the embryo during the recording may overwrite the perception of noniceptive stimuli. The results suggest developmental day 13 as the earliest embryonal stage being able to receive and process nociceptive stimuli.