Substance specific EEG patterns in mice undergoing slow anesthesia induction.

The exact mechanisms and the neural circuits involved in anesthesia induced unconsciousness are still not fully understood. To elucidate them valid animal models are necessary. Since the most commonly used species in neuroscience are mice, we established a murine model for commonly used anesthetics/sedatives and evaluated the epidural electroencephalographic (EEG) patterns during slow anesthesia induction and emergence. Forty-four mice underwent surgery in which we inserted a central venous catheter and implanted nine intracranial electrodes above the prefrontal, motor, sensory, and visual cortex. After at least one week of recovery, mice were anesthetized either by inhalational sevoflurane or intravenous propofol, ketamine, or dexmedetomidine. We evaluated the loss and return of righting reflex (LORR/RORR) and recorded the electrocorticogram. For spectral analysis we focused on the prefrontal and visual cortex. In addition to analyzing the power spectral density at specific time points we evaluated the changes in the spectral power distribution longitudinally. The median time to LORR after start anesthesia ranged from 1080 [1st quartile: 960; 3rd quartile: 1080]s under sevoflurane anesthesia to 1541 [1455; 1890]s with ketamine. Around LORR sevoflurane as well as propofol induced a decrease in the theta/alpha band and an increase in the beta/gamma band. Dexmedetomidine infusion resulted in a shift towards lower frequencies with an increase in the delta range. Ketamine induced stronger activity in the higher frequencies. Our results showed substance-specific changes in EEG patterns during slow anesthesia induction. These patterns were partially identical to previous observations in humans, but also included significant differences, especially in the low frequencies. Our study emphasizes strengths and limitations of murine models in neuroscience and provides an important basis for future studies investigating complex neurophysiological mechanisms.

Crosstalk between the subiculum and sleep-wake regulation: A review.

The circuitry underlying the initiation, maintenance, and coordination of wakefulness, rapid eye movement sleep, and non-rapid eye movement sleep is not thoroughly understood. Sleep is thought to arise due to decreased activity in the ascending reticular arousal system, which originates in the brainstem and awakens the thalamus and cortex during wakefulness. Despite the conventional association of sleep-wake states with hippocampal rhythms, the mutual influence of the hippocampal formation in regulating vigilance states has been largely neglected. Here, we focus on the subiculum, the main output region of the hippocampal formation. The subiculum, particulary the ventral part, sends extensive monosynaptic projections to crucial regions implicated in sleep-wake regulation, including the thalamus, lateral hypothalamus, tuberomammillary nucleus, basal forebrain, ventrolateral preoptic nucleus, ventrolateral tegmental area, and suprachiasmatic nucleus. Additionally, second-order projections from the subiculum are received by the laterodorsal tegmental nucleus, locus coeruleus, and median raphe nucleus, suggesting the potential involvement of the subiculum in the regulation of the sleep-wake cycle. We also discuss alterations in the subiculum observed in individuals with sleep disorders and in sleep-deprived mice, underscoring the significance of investigating neuronal communication between the subiculum and pathways promoting both sleep and wakefulness.

Altered sleep behavior strengthens face validity in the ArcAß mouse model for Alzheimer's disease.

Demographic changes will expand the number of senior citizens suffering from Alzheimer's disease (AD). Key aspects of AD pathology are sleep impairments, associated with onset and progression of AD. AD mouse models may provide insights into mechanisms of AD-related sleep impairments. Such models may also help to establish new biomarkers predicting AD onset and monitoring AD progression. The present study aimed to establish sleep-related face validity of a widely used mouse model of AD (ArcAß model) by comprehensively characterizing its baseline sleep/wake behavior. Chronic EEG recordings were performed continuously on four consecutive days in freely behaving mice. Spectral and temporal sleep/wake parameters were assessed and analyzed. EEG recordings showed decreased non-rapid eye movement sleep (NREMS) and increased wakefulness in transgenic mice (TG). Vigilance state transitions were different in TG mice when compared to wildtype littermates (WT). During NREMS, TG mice had lower power between 1 and 5 Hz and increased power between 5 and 30 Hz. Sleep spindle amplitudes in TG mice were lower. Our study strongly provides sleep-linked face validity for the ArcAß model. These findings extend the potential of the mouse model to investigate mechanisms of AD-related sleep impairments and the impact of sleep impairments on the development of AD.

An in-depth analysis of parameter settings and probability distributions of specific ordinal patterns in the Shannon permutation entropy during different states of consciousness in humans.

As electrical activity in the brain has complex and dynamic properties, the complexity measure permutation entropy (PeEn) has proven itself to reliably distinguish consciousness states recorded by the EEG. However, it has been shown that the focus on specific ordinal patterns instead of all of them produced similar results. Moreover, parameter settings influence the resulting PeEn value. We evaluated the impact of the embedding dimension m and the length of the EEG segment on the resulting PeEn. Moreover, we analysed the probability distributions of monotonous and non-occurring ordinal patterns in different parameter settings. We based our analyses on simulated data as well as on EEG recordings from volunteers, obtained during stable anaesthesia levels at defined, individualised concentrations. The results of the analysis on the simulated data show a dependence of PeEn on different influencing factors such as window length and embedding dimension. With the EEG data, we demonstrated that the probability P of monotonous patterns performs like PeEn in lower embedding dimension (m = 3, AUC = 0.88, [0.7, 1] in both), whereas the probability P of non-occurring patterns outperforms both methods in higher embedding dimensions (m = 5, PeEn: AUC = 0.91, [0.77, 1]; P(non-occurring patterns): AUC = 1, [1, 1]). We showed that the accuracy of PeEn in distinguishing consciousness states changes with different parameter settings. Furthermore, we demonstrated that for the purpose of separating wake from anaesthesia EEG solely pieces of information used for PeEn calculation, i.e., the probability of monotonous patterns or the number of non-occurring patterns may be equally functional.

Electroencephalographic monitoring of anesthesia during surgical procedures in mice using a modified clinical monitoring system.

Monitoring brain activity and associated physiology during the administration of general anesthesia (GA) in mice is pivotal to guarantee postanesthetic health. Clinically, electroencephalogram (EEG) monitoring is a well-established method to guide GA. There are no established methods available for monitoring EEG in mice (Mus musculus) during surgery. In this study, a minimally invasive rodent intraoperative EEG monitoring system was implemented using subdermal needle electrodes and a modified EEG-based commercial patient monitor. EEG recordings were acquired at three different isoflurane concentrations revealing that surgical concentrations of isoflurane anesthesia predominantly contained burst suppression patterns in mice. EEG suppression ratios and suppression durations showed strong positive correlations with the isoflurane concentrations. The electroencephalographic indices provided by the monitor did not support online monitoring of the anesthetic status. The online available suppression duration in the raw EEG signals during isoflurane anesthesia is a straight forward and reliable marker to assure safe, adequate and reproducible anesthesia protocols.

Nociception in Chicken Embryos, Part I: Analysis of Cardiovascular Responses to a Mechanical Noxious Stimulus.

Although it is assumed that chicken embryos acquire the capacity for nociception while developing in the egg, an exact time point has not yet been specified. The present research was an exploratory study aiming to determine when the capacity of nociception emerges during embryonic development in chickens. Changes in blood pressure and heart rate (HR) in response to a noxious mechanical stimulus at the base of the beak versus a light touch on the beak were examined in chicken embryos between embryonic days (EDs) 7 and 18. Mean arterial pressure (MAP) was the most sensitive parameter for assessing cardiovascular responses. Significant changes in MAP in response to a noxious stimulus were detected in embryos at ED16 to ED18, whereas significant changes in HR were observed at ED17 and ED18. Infiltration anesthesia with the local anesthetic lidocaine significantly reduced the response of MAP on ED18, so the measured cardiovascular changes may be interpreted as nociceptive responses.

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.

Nociception in Chicken Embryos, Part III: Analysis of Movements before and after Application of a Noxious Stimulus.

Many potentially noxious interventions are performed on chicken embryos in research and in the poultry industry. It is therefore essential and in the interest of animal welfare to be able to precisely define the point at which a chicken embryo is capable of nociception in ovo. The present part III of a comprehensive study examined the movements of developing chicken embryos with the aim of identifying behavioral responses to a noxious stimulus. For this purpose, a noxious mechanical stimulus and a control stimulus were applied in a randomized order. The recorded movements of the embryos were evaluated using the markerless pose estimation software DeepLabCut and manual observations. After the application of the mechanical stimulus, a significant increase in beak movement was identified in 15- to 18-day-old embryos. In younger embryos, no behavioral changes related to the noxious stimulus were observed. The presented results indicate that noxious mechanical stimuli at the beak base evoke a nocifensive reaction in chicken embryos starting at embryonic day 15.

Telemetric electroencephalography recording in anesthetized mice-A novel system using minimally-invasive needle electrodes with a wireless OpenBCI™ Cyton Biosensing Board.

Telemetric electroencephalography (EEG) recording, using subdermal needle electrodes, is a minimally-invasive method to investigate mammalian neurophysiology during anesthesia. These inexpensive systems may streamline experiments examining global brain phenomena during surgical anesthesia or disease. We utilized the OpenBCI™ Cyton board with subdermal needle electrodes to extract EEG features in six C57BL/6J mice undergoing isoflurane anesthesia. Burst suppression ratio (BSR) and spectral features were compared for a verification of our method. Following an increase from 1.5% to 2.0% isoflurane, the BSR increased (Wilcoxon-signed-rank statistic; p = 0.0313). Furthermore, although the absolute EEG spectral power decreased, the relative spectral power remained comparable (Wilcoxon-Mann-Whitney U-Statistic; 95% CI exclusive AUC=0.5; p < 0.05). Compared to tethered systems, this method confers several improvements for anesthesia specific protocols: 1-Avoiding electrode implant surgical procedures, 2-Anatomical non-specificity for needle electrode placement to monitor global cortical activity representative of anesthetic state, 3-Facility to repeat recordings in the same animal, 4-User-friendly for non-experts, 5-Rapid set-up time, and 6-Lower costs.•Minimally-invasive telemetric EEG recording systems ergonomically improve tethered systems for anesthesia protocols.•Using this method, we verified that higher isoflurane concentrations resulted in an increased EEG burst suppression ratio and decreased EEG absolute spectral power, with no change in frequency distribution.

Discriminating rapid eye movement sleep from wakefulness by analyzing high frequencies from single-channel EEG recordings in mice.

Rapid eye movement sleep (REMS) is characterized by the appearance of fast, desynchronized rhythms in the cortical electroencephalogram (EEG), similar to wakefulness. The low electromyogram (EMG) amplitude during REMS distinguishes it from wakefulness; therefore, recording EMG signal seems to be imperative for discriminating between the two states. The present study evaluated the high frequency components of the EEG signal from mice (80-500 Hz) to support REMS detection during sleep scoring without an EMG signal and found a strong positive correlation between waking and the average power of 80-120 Hz, 120-200 Hz, 200-350 Hz and 350-500 Hz. A highly negative correlation was observed with REMS. Furthermore, our machine learning approach demonstrated that simple EEG time-series features are enough to discriminate REMS from wakefulness with sensitivity of roughly 98 percent and specificity of around 92 percent. Interestingly, assessing only the higher frequency bands (200-350 Hz as well as 350-500 Hz) gives significantly greater predictive power than assessing only the lower end of the EEG frequency spectrum. This paper proposes an approach that can detect subtle changes in REMS reliably, and future unsupervised sleep-scoring approaches could greatly benefit from it.

Clinical and virological features of first human monkeypox cases in Germany.

BACKGROUND: Monkeypox is a zoonotic orthopoxvirus infection endemic in central and western Africa. In May 2022, human monkeypox infections including human-to-human transmission were reported in a multi-country outbreak in Europe and North America. CASE PRESENTATIONS: Here we present the first two cases of monkeypox infection in humans diagnosed in Germany. We present clinical and virological findings, including the detection of monkeypox virus DNA in blood and semen. The clinical presentation and medical history of our patients suggest close physical contact during sexual interactions as the route of infection. CONCLUSION: Monkeypox requires rapid diagnosis and prompt public health response. The disease should be considered in the current situation especially the differential diagnosis of vesicular or pustular rash, particularly in patients with frequent sexual contacts. Most importantly, it is essential to raise awareness among all health professionals for the rapid and correct recognition and diagnosis of this disease, which is probably still underreported in Europe (Adler et al. in Lancet Infect Dis https://doi.org/10.1016/s1473-3099(22)00228-6 , 2022).

Controlling absence seizures from the cerebellar nuclei via activation of the Gq signaling pathway.

Absence seizures (ASs) are characterized by pathological electrographic oscillations in the cerebral cortex and thalamus, which are called spike-and-wave discharges (SWDs). Subcortical structures, such as the cerebellum, may well contribute to the emergence of ASs, but the cellular and molecular underpinnings remain poorly understood. Here we show that the genetic ablation of P/Q-type calcium channels in cerebellar granule cells (quirky) or Purkinje cells (purky) leads to recurrent SWDs with the purky model showing the more severe phenotype. The quirky mouse model showed irregular action potential firing of their cerebellar nuclei (CN) neurons as well as rhythmic firing during the wave of their SWDs. The purky model also showed irregular CN firing, in addition to a reduced firing rate and rhythmicity during the spike of the SWDs. In both models, the incidence of SWDs could be decreased by increasing CN activity via activation of the Gq-coupled designer receptor exclusively activated by designer drugs (DREADDs) or via that of the Gq-coupled metabotropic glutamate receptor 1. In contrast, the incidence of SWDs was increased by decreasing CN activity via activation of the inhibitory Gi/o-coupled DREADD. Finally, disrupting CN rhythmic firing with a closed-loop channelrhodopsin-2 stimulation protocol confirmed that ongoing SWDs can be ceased by activating CN neurons. Together, our data highlight that P/Q-type calcium channels in cerebellar granule cells and Purkinje cells can be relevant for epileptogenesis, that Gq-coupled activation of CN neurons can exert anti-epileptic effects and that precisely timed activation of the CN can be used to stop ongoing SWDs.

The impact of tethered recording techniques on activity and sleep patterns in rats.

Electrophysiological recordings in animals constitute frequently applied techniques to study neuronal function. In this context, several authors described tethered recordings as a semi-restraint situation with negative implications for animal welfare and suggested radiotelemetric setups as a refinement measure. Thus, we here investigated the hypothesis that tethered recordings exert measurable effects on behavioral and sleep patterns in Sprague-Dawley rats. Animals were kept in monitoring glass cages either with or without a head connection to a recording cable. Saccharin preference, nest building, serum corticosterone and fecal corticosterone metabolite levels were in a comparable range in both groups. The proportion of vigilance states was not affected by the cable connection. Minor group differences were detected in bout lengths distributions, with a trend for longer NREM and WAKE stages in animals with a cable connection. However, a relevant effect was not further confirmed by an analysis of the number of sleep/wake and wake/sleep transitions. The analysis of activity levels did not reveal group differences. However, prolonged exposure to the tethered condition resulted in an intra-group increase of activity. In conclusion, the comparison between freely moving vs tethered rats did not reveal major group differences. Our findings indicate that telemetric recordings only offer small advantages vs cabled set ups, though this may differ in other experimental studies where for example anxiety- or drug-induced effects are analyzed.

Altered sleep behavior in a genetic mouse model of impaired fear extinction.

Sleep disturbances are a common complaint of anxiety patients and constitute a hallmark feature of post-traumatic stress disorder (PTSD). Emerging evidence suggests that poor sleep is not only a secondary symptom of anxiety- and trauma-related disorders but represents a risk factor in their development, for example by interfering with emotional memory processing. Fear extinction is a critical mechanism for the attenuation of fearful and traumatic memories and multiple studies suggest that healthy sleep is crucial for the formation of extinction memories. However, fear extinction is often impaired in anxiety- and trauma-related disorders-an endophenotype that is perfectly modelled in the 129S1/SvImJ inbred mouse strain. To investigate whether these mice exhibit altered sleep at baseline that could predispose them towards maladaptive fear processing, we compared their circadian sleep/wake patterns to those of typically extinction-competent C57BL/6 mice. We found significant differences regarding diurnal distribution of sleep and wakefulness, but also sleep architecture, spectral features and sleep spindle events. With regard to sleep disturbances reported by anxiety- and PTSD patients, our findings strengthen the 129S1/SvImJ mouse models' face validity and highlight it as a platform to investigate novel, sleep-focused diagnostic and therapeutic strategies. Whether the identified alterations causally contribute to its pathological anxiety/PTSD-like phenotype will, however, have to be addressed in future studies.

Combined implanted central venous access and cortical recording electrode array in freely behaving mice.

Establishing a long-lasting, functioning venous access in a non-anesthetized mouse is very challenging at least. Since we needed a reliable venous access to titrate intravenous anesthetics, we refined and combined previously described methods. The tunneling of the catheter from the cranial to the pectoral wound, the fixation of the catheter in the external jugular vein with two sutures, and a tissue adhesive allowed us to combine this method with the implantation of intracranial recording electrodes. With this approach we neither have to restrain the animal causing excessive stress nor do we need an additional anesthetic, interfering with the effects of the intravenous anesthetic. This approach can help to establish a greater understanding of the concept of consciousness by identifying the neural circuits which mediate the effect of intravenous anesthetics. In addition - due to the flexible design of the recording electrode array - our approach can also be applied to investigate further neuroscientific hypotheses.•Establishment of a reliable chronical venous access for the application in freely behaving mice.•The jugular venous access can be combined with all kinds of neurobiological recording and application designs.•The design of the venous access allows chronic combinations with telemetric and tether-bound systems.

Sleep/Wake Behavior and EEG Signatures of the TgF344-AD Rat Model at the Prodromal Stage.

Transgenic modification of the two most common genes (APPsw, PS1ΔE9) related to familial Alzheimer's disease (AD) in rats has produced a rodent model that develops pathognomonic signs of AD without genetic tau-protein modification. We used 17-month-old AD rats (n = 8) and age-matched controls (AC, n = 7) to evaluate differences in sleep behavior and EEG features during wakefulness (WAKE), non-rapid eye movement sleep (NREM), and rapid eye movement sleep (REM) over 24-h EEG recording (12:12h dark-light cycle). We discovered that AD rats had more sleep-wake transitions and an increased probability of shorter REM and NREM bouts. AD rats also expressed a more uniform distribution of the relative spectral power. Through analysis of information content in the EEG using entropy of difference, AD animals demonstrated less EEG information during WAKE, but more information during NREM. This seems to indicate a limited range of changes in EEG activity that could be caused by an AD-induced change in inhibitory network function as reflected by increased GABAAR-ß2 expression but no increase in GAD-67 in AD animals. In conclusion, this transgenic rat model of Alzheimer's disease demonstrates less obvious EEG features of WAKE during wakefulness and less canonical features of sleep during sleep.

Cyto- and myeloarchitectural brain atlas of the pale spear-nosed bat (Phyllostomus discolor) in CT Aided Stereotaxic Coordinates.

The pale spear-nosed bat Phyllostomus discolor, a microchiropteran bat, is well established as an animal model for research on the auditory system, echolocation and social communication of species-specific vocalizations. We have created a brain atlas of Phyllostomus discolor that provides high-quality histological material for identification of brain structures in reliable stereotaxic coordinates to strengthen neurobiological studies of this key species. The new atlas combines high-resolution images of frontal sections alternately stained for cell bodies (Nissl) and myelinated fibers (Gallyas) at 49 rostrocaudal levels, at intervals of 350 µm. To facilitate comparisons with other species, brain structures were named according to the widely accepted Paxinos nomenclature and previous neuroanatomical studies of other bat species. Outlines of auditory cortical fields, as defined in earlier studies, were mapped onto atlas sections and onto the brain surface, together with the architectonic subdivisions of the neocortex. X-ray computerized tomography (CT) of the bat's head was used to establish the relationship between coordinates of brain structures and the skull. We used profile lines and the occipital crest as skull landmarks to line up skull and brain in standard atlas coordinates. An easily reproducible protocol allows sectioning of experimental brains in the standard frontal plane of the atlas. An electronic version of the atlas plates and supplementary material is available from https://doi.org/10.12751/g-node.8bbcxy.

Teacher Procrastination, Emotions, and Stress: A Qualitative Study.

Stress and negative emotions in teachers can lead to occupational burnout, poor performance in the classroom, and decreased job-satisfaction. Apart from having negative personal and physical effects for the individual teacher, teacher stress and burnout are also thought to have negative effects on the respective students and student achievement. As one potential source of teacher stress, procrastination has been speculated about. However, research on the phenomenology and prevalence of procrastination among teachers, as well as its relevance for their emotional and stress experiences, is very scarce. Further, most of the existing research on teacher procrastination used general self-report scales to obtain results. The present study therefore investigated the phenomenology of teacher procrastination as well as its links with emotional experiences and stress, using a qualitative approach. Twenty-seven male and female teachers from Germany were interviewed personally (Mean age = 35.7, SD = 9.64, Min = 25 years, Max = 67 years). Nine of those teachers reported to never needlessly delay an action concerning their profession or not to perceive their dilatory behavior as negative and stressful. Data from the remaining 16 teachers (Mean age = 35.06, SD = 7.01, Min = 26 years, Max = 48 years) were analyzed on the basis of qualitative content analysis by using deductive as well as inductive category application. Results revealed that these teachers procrastinate on an array of professional tasks, such as administrative and organizational tasks and correcting students' work. The results showed that teachers delayed these tasks for different reasons but mainly due to task aversiveness. Further, teachers reported experiencing mainly negative emotions when procrastinating and perceiving their procrastination behavior as moderately stressful, indicating that procrastination is a potential stressor in the teacher profession. Limitations of the study are discussed and directions for future research are proposed.

Distinct Parameters in the EEG of the PLP α-SYN Mouse Model for Multiple System Atrophy Reinforce Face Validity.

Multiple system atrophy (MSA) is a neurodegenerative movement disorder characterized by parkinsonian symptoms and cerebellar symptoms. Sleep disturbances also play a crucial role in MSA. One of the most convincing animal models in MSA research is the PLP α-SYN model, but to date no studies on sleep disturbances in this mouse model, frequently found in MSA patients are available. We identified spectral shifts within the EEG of the model, strikingly resembling results of clinical studies. We also characterized muscle activity during REM sleep, which is one of the key symptoms in REM sleep behavioral disorder. Spectral shifts and REM sleep-linked muscle activity were age dependent, supporting Face Validity of the PLP α-SYN model. We also strongly suggest our findings to be critically evaluated for Predictive Validity in future studies. Currently, research on MSA lacks potential compounds attenuating or curing MSA. Future drugs must prove its potential in animal models, for this our study provides potential biomarkers.

The khmer software package: enabling efficient nucleotide sequence analysis.

The khmer package is a freely available software library for working efficiently with fixed length DNA words, or k-mers. khmer provides implementations of a probabilistic k-mer counting data structure, a compressible De Bruijn graph representation, De Bruijn graph partitioning, and digital normalization. khmer is implemented in C++ and Python, and is freely available under the BSD license at  https://github.com/dib-lab/khmer/.