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The ability to learn in the context of predation allows prey to respond to threats by adjusting their behavior based on specific information acquired from their current environment. Habituation is a process that allows animals to adapt to environmental changes. Very little is known about habituation in wild animals in general and there are no studies on habituation in anuran tadpoles in particular. Here, we performed three experiments to investigate the behavioral response of predator naïve Pleurodema thaul tadpoles to repeated stimulation with two predation risk cues (injured conspecific and predator fed cues) which a priori provide different information regarding risk. Experiment 1 showed that P. thaul tadpoles habituate the antipredator response when undergo predation risk chemical cues from injured conspecific and that response is long term. Experiment 2 showed that P. thaul tadpoles did not habituate their antipredator response when exposed to cues derived from an event of nymph odonate preying on P. thaul tadpoles (predator fed cues). Experiment 3 specifically evaluated the risk imposed by each of the risk cues used in Experiment 1 and Experiment 2 and showed that the degree of perceived risk in tadpoles appear to be similar in a single experience with any risk stimuli. We suggest that the behavioral habituation of tadpoles in the context of predation could be modulated by the level of uncertainty associated with risk stimuli.

Summary Complex behavioral phenotyping techniques are becoming more prevalent in the field of behavioral neuroscience, and thus methods for manipulating neuronal activity must be adapted to fit into such paradigms. Here, we present a head-mounted, magnetically activated device for wireless optogenetic manipulation that is compact, simple to construct, and suitable for use in group-living mice in an enriched semi-natural arena over several days. Using this device, we demonstrate that repeated activation of oxytocin neurons in male mice can have different effects on pro-social and agonistic behaviors, depending on the social context. Our findings support the social salience hypothesis of oxytocin and emphasize the importance of the environment in the study of social neuromodulators. Our wireless optogenetic device can be easily adapted for use in a variety of behavioral paradigms, which are normally hindered by tethered light delivery or a limited environment., Graphical Abstract, Highlights • A small, wireless device is used for optogenetic activation in a complex environment • PVN oxytocin neurons were activated repeatedly over 2 days in a group setting • Repeated activation in a group setting elicited both pro-social and agonistic behavior • Findings support the social salience hypothesis of oxytocin neuro-modulation, Anpilov et al. present a lightweight, easily assembled wireless optogenetic device compatible with group-housed mice in a complex environment. Using this approach, they show that repeated activation of oxytocin neurons elicits both pro-social and aggressive behaviors over time, emphasizing the role of the social setting in modulating behavior.

Key points To explore the capability of cathodal transcranial direct current stimulation (tDCS) to induce late-phase long-term depression (LTD) via repeated stimulation. Conventional (1 mA for 15 min) and intensified (3 mA for 20 min) protocols with short (20 min) and long (24 h) intervals were tested. Late-phase plasticity was not induced by a single repetition of stimulation. Repetition reduced the efficacy of stimulation protocols with higher intensities. Abstract Transcranial direct current stimulation (tDCS) has shown promising results in pilot studies as a therapeutic intervention in disorders of the central nervous system, but more sustained effects are required for clinical application. To address this issue, one possible solution is the use of repeated stimulation protocols. Previous studies indicated the possibility of extending the after-effects of single intervention cathodal tDCS by repeating the tDCS, with relatively short intervals between repetitions being most effective. In this study, we thus investigated the effects of repeated stimulation protocols at short and long intervals, for a conventional tDCS protocol (1 mA for 15 min) and a newly developed optimized protocol (3 mA for 20 min). In 16 healthy participants, we compared single interventions of conventional and optimized protocols, repeated application of these protocols at intervals of 20 min and 24 h, and a sham tDCS session. tDCS-induced neuroplastic after-effects were then monitored with transcranial magnetic stimulation (TMS)-induced motor evoked potentials (MEPs) until the following evening after stimulation. The results revealed that the duration of the after-effects of repeated conventional and optimized protocols with short intervals remained nearly unchanged compared to the respective single intervention protocols. For the long-interval (24 h) protocol, stimulation with the conventional protocol did not significantly alter respective after-effects, while it reduced the efficacy of the optimized protocol, compared with respective single interventions. Thus late-phase plasticity could not be induced by a single repetition of stimulation in this study, but repetition reduced the efficacy of stimulation protocols with higher intensities. This study provides further insights into the dependency of tDCS-induced neuroplasticity on stimulation parameters, and therefore delivers crucial information for future tDCS applications.

The purpose of this review article is to summarize our current understanding of the efficacy and safety of cardiac defibrillation with nanosecond shocks. Experiments in isolated hearts, using optical mapping of the electrical activity, have demonstrated that nanosecond shocks can defibrillate with lower energies than conventional millisecond shocks. Single defibrillation strength nanosecond shocks do not cause obvious damage, but repeated stimulation leads to deterioration of the hearts. In isolated myocytes, nanosecond pulses can also stimulate at lower energies than at longer pulses and cause less electroporation (propidium uptake). The mechanism is likely electroporation of the plasma membrane. Repeated stimulation leads to distorted calcium gradients.

Two thirds of the 302 neurons in C. elegans form bilaterally symmetric pairs in its physical connectome, and similar gross morphological symmetries are seen in the nervous systems of many other animals. A central question is whether and how this morphological symmetry is broken to produce functional asymmetry. Addressing this question, in all but two cases, has been impossible because no promoters are known that can direct gene expression to a single cell within a symmetric pair. Here we develop an efficient genetic code expansion system in C. elegans and use this system to create a photo-activatable version of Cre recombinase. Using this system, we target single neurons within a bilaterally symmetric pair (PLMR and PLML) with a laser. This turns on Cre and thereby switches on expression of an optogenetic channel in a single cell. We hereby overcome the limitation that these neurons cannot be targeted by genetic means. Our approach enables the generation of large numbers of animals for downstream experiments. By globally illuminating groups of freely moving animals to stimulate the targeted neurons that express an optogenetic channel we dissect the individual contributions of PLMR and PLML to the C. elegans touch response. Our results reveal that the individual neurons make asymmetric contributions to this behaviour, and suggest distinct roles for PLMR and PLML in the habituation to repeated stimulation. Our results demonstrate how genetic code expansion and optical targeting can be combined to break the symmetry of neuron pairs in C. elegans and thereby dissect the contributions of individual neurons to behaviour.

How the body is represented in the developing brain is a topic of growing interest. The current study takes a novel approach to investigating neural body representations in infants by recording somatosensory mismatch negativity (sMMN) responses elicited by tactile stimulation of different body locations. Recent research in adults has suggested that sMMN amplitude may be influenced by the relative distance between representations of the stimulated body parts in somatosensory cortex. The current study uses a similar paradigm to explore whether the sMMN can be elicited in infants, and to test whether the infant sMMN response is sensitive to the somatotopic organization of somatosensory cortex. Participants were healthy infants (n = 31) aged 6 and 7 months. The protocol leveraged a discontinuity in cortical somatotopic organization, whereby the representations of the neck and the face are separated by representations of the arms, the hands and the shoulder. In a double-deviant oddball protocol, stimulation of the hand (100 trials, 10% probability) and neck (100 trials, 10% probability) was interspersed among repeated stimulation of the face (800 trials, 80% probability). Waveforms showed evidence of an infant sMMN response that was significantly larger for the face/neck contrast than for the face/hand contrast. These results suggest that, for certain combinations of body parts, early pre-attentive tactile discrimination in infants may be influenced by distance between the corresponding cortical representations. The results provide the first evidence that the sMMN can be elicited in infants, and pave the way for further applications of the sMMN in studying body representations in preverbal infants.

Heartburn is one of the most widely spread symptoms particularly in population of economically developed Western countries. Traditionally it is considered to be related to activation of chemo- and mechanoreceptors of esophageal primary sensory neurons. Repeated stimulation may provoke visceral sensitization. Its development is facilitated by the stress hormone corticotropin-releasing factor. An attentive analysis of heartburn with an assessment of the provoking factors and effectiveness of drug therapy is required for differentiation of gastro-intestinal reflux disease and functional esophageal disease. The lack of response to proton pump inhibitors is an important criterion for suspicion on functional esophageal pathology although sensitization also plays an important role in genesis of heartburn in non-erosive reflux disease. In cases of refractory and «endoscopically negative heartburn», additional studies are justified: endoscopy with esophageal biopsies, 24h-pH-metry/pH-impedansometry, high-resolution manometry. Proton pump inhibitors, highly effective in reflux disease, may relieve heartburn only in some cases of functional heartburn and reflux hypersensitivity. In functional diseases medical therapy affecting visceral hypersensitivity may be used empirically.

Excessive participation in casual leisure activities often leads to satiation, which leaves people no pleasure. Prior research did not consider the gradation effects on the accumulation of satiation. This study proposed a theoretical model to illuminate the changing process of satiation. With repeated stimulation, people undergo three stages: implicit satiation (S1), explicit satiation (S2), and dull satiation (S3). This study conceived the idea of satiation stages. The changing process of enjoyment was studied as the participants listened to repeated music during the experiments. Results show that when total utility is used as a criterion, the marginal utility of stage S1 yields higher affective experience than that of stage S2 and S3. Negative emotions cause individuals to adopt the avoidant coping strategy, S3, which involves more repetition, thereby resulting in a better experience memory and taking less recovery time than S2.

Orienting responses (ORs) are whole-organism reflexes that are elicited by innocuous stimuli, and which decrease in magnitude after stimulus repetition. ORs represent relatively simple responses that can be used to study attentional processes, and are modulated by the organism’s state, including arousal and activation levels, as well as by emotional processes. Here we describe a simple method to study ORs in zebrafish, a model organism increasingly being used in behavioural neuroscience. After presentation of a static visual stimulus, an OR is elicited, characterized by approaching the stimulus and orienting towards it. After repeated stimulation, OR decreases, suggesting habituation. These responses are qualitatively altered by exposure to a fear-eliciting alarm substance (i.e., derived from the skin of a conspecific), since exposed animals avoid the visual stimulus and orient either away from the stimulus or towards it, but at a distance. The protocol can be used to study orienting responses, as well as the impact of fear and arousal on these reflexes.

Objective. Stimulator-attached dissecting instruments are useful for intraoperative nerve monitoring during thyroidectomy. The aim of this study was to evaluate the feasibility of an attachable ring stimulator (ARS) by comparing the electromyography (EMG) amplitudes evoked by an ARS and a conventional stimulator. Methods. Medical records of fourteen patients who underwent thyroidectomy using intraoperative neuromonitoring between June and August 2019 were retrospectively reviewed. The amplitudes of V1, R1, R2, and V2 signals were checked using both the ARS and a conventional stimulator, at the same point. Results. Both stimulators were tested on 20 recurrent laryngeal nerves (RLNs) and 20 vagus nerves (VNs). In all the nerves, the amplitudes of V1, R1, R2, and V2 were greater than 500 μV. The mean amplitudes of V1, R1, R2, and V2 checked with the ARS were 1175, 1432, 1598, and 1279 μV, respectively. The mean amplitudes of V1, R1, R2, and V2 checked with the conventional stimulator were 1140, 1425, 1557, and 1217 μV, respectively. Difference between amplitudes evoked by the two stimulators for V1, R1, R2, and V2 was 77, 110, 102, and 99 μV, respectively. There was no statistical difference in the amplitudes between the two groups for V1, R1, R2, and V2. Conclusion. The ARS transferred electric stimulation as effectively as the conventional stimulator. It is an effective tool for repeated stimulation and facilitates continuous feedback regarding the functional integrity of nerves during thyroid surgery.

We here provide an update about studies published recently in the field of noninvasive neuromodulation of the motor system, aiming at facilitating recovery of function after stroke.A number of longitudinal studies have confirmed that repeated stimulation of the motor cortex in combination with motor training improves performance compared with control or sham stimulation. In the early postacute stroke phase, enhancement of ipsilesional motor cortex excitability by means of repetitive transcranial magnetic stimulation (rTMS) seems to be a well tolerated and effective strategy to promote motor recovery. In contrast, recent studies suggest that transcranial direct current stimulation (tDCS) in the early poststroke phase does not facilitate motor recovery. In the chronic phase, however, both rTMS and tDCS have been shown to be beneficial when applied over several days combined with training. Interestingly, bihemispheric stimulation strategies (ipsilesional activation and contralesional suppression) seem to constitute effective protocols, especially when using rTMS.Noninvasive brain stimulation seems to support motor recovery. However, to date randomised controlled trials (RCTs) that are sufficiently powered are lacking. Data suggest that the most promising protocols should now be tested in RCTs with sufficiently large samples taking into account the clinical heterogeneity of stroke.

Gagliano et al. (Oecologia 175(1):63-72, 2014) reported that Mimosa pudica habituates to repeated stimulation, as shown by a reduction in response, dishabituation, and stimulus specificity. I argue that Gagliano et al.'s data show an absence of dishabituation, that their experimental design needs an additional condition to test whether there is stimulus specificity, and that most of their data can be explained by motor fatigue. Some data are not easily explained by fatigue, and I suggest a further analysis that may clarify the issue. The status of habituation in Mimosa remains uncertain.

One of the areas where the concept of hormesis has rapidly gained acceptance is in aging research and interventions. Mild stress-induced repeated stimulation of protective mechanisms in cells and organisms has a wide range of health-promoting and life span-extending effects. Moderate and repeated physical exercise is the paradigm for the health beneficial and aging-modulatory effects of hormesis. Molecular mechanisms for the action of hormetins, which bring about the hormesis, comprise a cascade of stress response pathways and stimulation of other pathways of maintenance and repair. Hormetic strengthening of homeodynamics provides wider margins for metabolic fluctuation, stress tolerance, adaptation, and survival of the organism. Hormesis thus counterbalances the progressive shrinkage of the homeodynamic space, which is the major phenotype of aging.

New findings What is the central question of this study? Is the residual force generated by the diaphragm muscle after repeated activation reduced with sarcopenia, and is the residual force generated after fatiguing activation sufficient to sustain ventilatory behaviours of diaphragm muscle in young and old rats? What is the main finding and its importance? After diaphragm muscle fatigue, the residual specific force after 120 s of repeated stimulation was unaffected by ageing and was sufficient to accomplish ventilatory behaviours, but not expulsive manoeuvres (e.g. coughing). The inability to perform expulsive behaviours might underlie the increased susceptibility of older individuals to respiratory tract infections. Abstract Type IIx and/or IIb diaphragm muscle (DIAm) fibres make up more fatigable motor units that are more vulnerable to sarcopenia, i.e. age-associated reductions of specific force and cross-sectional area. In contrast, type I and IIa DIAm fibres form fatigue-resistant motor units that are relatively unchanged with age. The fatigue resistance of the DIAm is assessed by normalizing the residual force generated after a period of repeated supramaximal stimulation (e.g. 120 s) to the initial maximal force. Given that sarcopenia primarily affects more fatigable DIAm motor units, apparent fatigue resistance improves with ageing. However, the central question is whether there is an ageing-related difference in the residual force generated by the DIAm after repeated stimulation and whether this force is sufficient to sustain ventilatory behaviours of DIAm. In 6- and 24-month-old Fischer 344 rats, we assessed the loss of ex vivo DIAm force throughout 120 s of repeated supramaximal stimulation at 10, 40 and 75 Hz. We found that relative fatigue resistance improved in older rats at 40 and 75 Hz stimulation. Across all stimulation frequencies, DIAm residual force was unchanged with age (∼5 N cm-2 ). We conclude that ageing increases the relative contribution of type I and IIa fibres to DIAm force, with decreased contributions of type IIx and/or IIb fibres. The residual force generated by the DIAm after repeated stimulation is sufficient to accomplish ventilatory behaviours, regardless of age.

Gynecomastia consists of breast enlargement due to a hormonal imbalance between estrogens and androgens. Unilateral and important breast growth requires ruling underlying pathologic disorders out. Mechanical cause is uncommon, causing enlargement by repeated stimulation. We report a 6-year-old boy with unilateral gynecomastia. Hyperprolactinemia is the only abnormal finding at laboratory tests. After repeated inquiries, a continuous breast selfstimulation is detected. Its relation with gynecomastia is verified because prolactin normalizes and breast regressed in further revisions, after stopping stimulus.

Uniquely expressing diverse innate-like and adaptive-like functions, γδ T cells exist as specialized subsets, but are also able to adapt in response to environmental cues. These cells have long been known to rapidly proliferate following primary malaria infection in humans and mice, but exciting new work is shedding light into their diverse functions in protection and following repeated malaria infection. In this review, we examine the current knowledge of functional specialization of γδ T cells in malaria, and the mechanisms dictating recognition of malaria parasites and resulting proliferation. We discuss γδ T cell plasticity, including changing interactions with other immune cells during recurrent infection and potential for immunological memory in response to repeated stimulation. Building on recent insights from human and murine experimental studies and vaccine trials, we propose areas for future research, as well as applications for therapeutic development.

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for erectile dysfunction. However, the methods used vary between laboratories, and pitfalls still exist. The goal of this study was to describe a surgical technique that would provide a reliable and reproducible model. By exposing the ischiocavernosus muscle at its point of insertion on the ischial tuberosity, the penile crus could be cannulated with minimal dissection and injury to the structures involved in erectile function. Repeated stimulation of the CN, without the need for lifting and drying, was achieved by using a 125 µm bipolar silver electrode and biocompatible silicon glue to isolate the electrode-nerve complex. This method prevents neuropraxia by reducing stretching and drying the nerve and provides complete isolation of the nerve, negating electrical leakage and preventing stimulation of alternative pathways.

There has been long-standing speculation that autism spectrum disorder (ASD) involves an increase in excitation relative to inhibition. However, there is little direct evidence of increased neural excitation in humans with ASD. Here we provide a potential explanation for this discrepancy: we show that increased neural excitation emerges only after repeated stimulation, manifesting as a deficit in neural adaptation. We measured fMRI responses induced by repeated audio-visual stimulation and button presses in early sensory-motor cortical areas. Across all cortical areas we show reduced adaptation in individuals with ASD compared to neurotypical individuals. The degree of adaptation is correlated between cortical areas and with button-press reaction times across subjects. These findings suggest that increased neural excitation in ASD, manifesting as dysregulated neural adaptation, is domain-general and behaviorally-relevant.

Hagfishes are known for their unique defensive slime, which they use to ward off gill-breathing predators. Although much is known about the slime cells (gland thread cells and gland mucous cells), little is known about how long slime gland refilling takes, or how slime composition changes with refilling or repeated stimulation of the same gland. Slime glands can be individually electrostimulated to release slime, and this technique was used to measure slime gland refilling times for Atlantic and Pacific hagfish. The amount of exudate produced, the composition of the exudate and the morphometrics of slime cells were analyzed during refilling, and as a function of stimulation number when full glands were stimulated in rapid succession. Complete refilling of slime glands for both species took 3–4 weeks, with Pacific hagfish achieving faster absolute rates of exudate recovery than Atlantic hagfish. We found significant changes in the composition of the exudate and in the morphometrics of slime cells from Pacific hagfish during refilling. Over successive stimulations of full Pacific hagfish glands, multiple boluses of exudate were released, with exudate composition, but not thread cell morphometrics, changing significantly. Finally, histological examination of slime glands revealed slime cells retained in glands after exhaustion. Discrepancies in the volume of cells released suggest that mechanisms other than contraction of the gland musculature alone may be involved in exudate ejection. Our results provide a first look at the process and timing of slime gland refilling in hagfishes, and raise new questions about how refilling is achieved at the cellular level.

In rodent's olfactory bulb, different odors evoke distinct glomeruli patterns to form a specific ‘Odor Map’. The optical imaging has the ability to visualize the functional responses in precise spatial and temporal resolution, which provides an opportunity to decipher the functional architecture of the patterns in the olfactory bulb. Here we adapted intrinsic signal imaging to visualize the odor response in the olfactory bulb. The imaging signals displayed clear odor-evoked pattern with stability over repeated stimulation. The imaging results also revealed a more focal response in comparison with electrophysiological activity. These characteristics suggest the possibility to use the optical imaging signals as the input of a brain-machine interface.

The cardiac defense response (CDR) to intense auditory stimulation is characterized by two acceleration-deceleration heart rate (HR) components. This study investigated contributions of sympathetic cardiac control to habituation and recovery of the CDR. Fifty-six healthy subjects were presented with noise stimuli eliciting the CDR. Three stimuli were presented with short and long (2.5 min and 12.5 min) inter-trial intervals (ITIs). The pre-ejection period was recorded as an index of sympathetic cardiac control, in addition to HR. Repeated stimulation at short ITI was associated with marked habituation of the HR and sympathetic responses; both responses exhibited a degree of recovery with long ITI. Regarding the time course, the first acceleration-deceleration was accompanied by a decline and subsequent increase in sympathetic cardiac control. During the second acceleration-deceleration, the parameters exhibited parallel courses. These results suggest that the sympathetic contribution to the habituation and recovery is limited to the second HR component.

Repetitive transcranial magnetic stimulation (rTMS) is emerging as a valuable adjunctive therapeutic modality in movement disorders. It is a non-invasive technique of repeated stimulation of the cerebral cortex by a train of magnetic pulses. The therapeutic effect of rTMS was first noted in depression. Later several researchers have investigated the role of rTMS in various movement disorders, notably Parkinson's disease, dystonia, Tourette's syndrome etc. The rTMS protocols used in these studies vary widely, lacks uniformity and often the results are not consistent. The optimal rTMS parameters for each disorder are yet to be established. This review discusses the current knowledge on the therapeutic applications of rTMS in various movement disorders.

Adrenal myelolipomas are rare benign tumors of the adrenal cortex composed of adipose and hematopoietic cells. They have been postulated to arise from repeated stimulation by stress, inflammation and ACTH oversecretion. Myelolipomas are usually detected incidentally on imaging and do not require any active intervention besides regular follow-up by imaging. However, myelolipomas may insidiously grow to large sizes and cause mass effects and hemorrhage. Timely diagnosis and surgical resection are curative and lifesaving.

Habituation, or the relatively permanent waning of a response as a result of repeated stimulation, is a form of behavioural plasticity that allows animals to filter out irrelevant stimuli and to focus selectively on important stimuli. Individuals that fail to habituate might be at a disadvantage if they continue to respond to irrelevant stimuli; therefore, habituation can have adaptive significance. In this study we compared rates of behaviour over time toward three different ecologically-relevant stimuli (food, a male intruder and a gravid female) in threespine sticklebacks (Gasterosteus aculeatus). We detected evidence for habituation to the stimuli, and males in this study were especially aggressive toward both male and female conspecifics. Although there were some clear temporal patterns that could be detected by looking at average behaviour, not all individuals behaved in the same ‘average’ way. We detected substantial inter-individual variation in behaviour toward all three stimuli, inter-individual variation in rates of habituation to both male and female conspecifics, but no evidence for correlations between behaviours across stimuli (behavioural syndromes). These results suggest that individual animals vary in rates of habituation, and prompt hypotheses about the causes and consequences of variation in rates of habituation.

We studied the effects of repeated stimulation by recombinant human FSH (rhFSH) at various time intervals during a physiologic breeding season in rhesus monkeys. Ovarian recovery and responses were assessed by ultrasonography, serum steroid concentrations, number of oocytes retrieved, and in vitro blastocyst development following IVF. One group underwent a single stimulation regimen with 18 IU rhFSH i.m., followed by 1000 IU hCG, and serum steroid concentrations and ovarian status were determined in the following three menses. Another group was stimulated as before and then allocated into three subgroups; each subgroup was re-stimulated once at the beginning of the ensuing first, second, or third menses. In the final experiment, one group was stimulated with 37.5 IU rhFSH, whereas another group received 18 IU rhFSH. In subsequent cycles, all were re-stimulated twice with 18 IU rhFSH at time intervals of two menstrual cycles (MCs). At the first menses after stimulation, serum progesterone concentrations were significantly higher and the ovaries larger than before stimulation. Monkeys that were re-stimulated at the first menses responded poorly; at the second menses, progesterone concentrations and ovarian size recovered, but the number of oocytes retrieved from re-stimulated monkeys was still significantly reduced. However, animals that were re-stimulated in two MCs later responded well (i.e., percentage of the animals responding, oocytes recovered, and potential for fertilization and blastocyst formation). In conclusion, rhesus monkeys were likely to have similar ovarian responses to repeated stimulation with the same regimen spaced at least two MCs apart.

Microelectrode arrays (MEAs) are used to study the electrical activity in brain slices and neuronal cultures. MEA experiments for the analysis of electrical stimulation responses require the tissue or culture to be prone to stimulation. For brain slices, potential stimulation sites may be directly visible in microscope, in which case the determination of stimulability at those locations is sufficient. In unstructured neuronal cultures, potential stimulation sites may not be known a priori, and spatial stimulability screening should be performed. Considering, e.g., 59 microelectrode sites, each to be stimulated several times, may result in long screening times, unacceptable with a MEA system without an integrated CO2 incubator, or in high stimulation effects on the networks. Here, we describe an implementation of a fast stimulation protocol employing pseudorandom stimulation site switching aiming at alleviating the network effects of the stimulability screening. In this paper, we show the usability of the proposed protocol by first detecting stimulable locations and subsequently apply repeated stimulation on the identified potentially stimulable locations to observe an exemplary neuronal pathway.

Background Habituation, as a basic form of learning, is characterized by decreasing amplitudes of neuronal reaction following repeated stimuli. Recent studies indicate that habituation to pure tones of different frequencies occurs in fetuses and infants. Aims Neural processing of different syllables in fetuses and infants was investigated. Study design An auditory habituation paradigm including two different sequences of syllables was presented to each subject. Each sequence consisted of eight syllables (sequence /ba/: 5 × /ba/, 1 × /bi/ (dishabituator), 2 × /ba/; sequence /bi/: 5 × /bi/, 1 × /ba/ (dishabituator), 2 × /bi/). Each subject was stimulated with 140 sequences. Neuromagnetic signatures of auditory-evoked responses (AER) were recorded by fetal magnetoencephalography (fMEG). Subjects Magnetic brain signals of N = 30 fetuses (age: 28–39 weeks of gestation) and N = 28 infants (age: 0–3 months) were recorded. Forty-two of the 60 fetal recordings and 29 of the 58 infant recordings were included in the final analysis. Outcome measures AERs were recorded and amplitudes were normalized to the amplitude of the first stimulus. Results In both fetuses and infants, the amplitudes of AERs were found not to decrease with repeated stimulation. In infants, however, amplitude of syllable 6 (dishabituator) was significantly increased compared to syllable 5 (p = 0.026). Conclusions Fetuses and infants showed AERs to syllables. Unlike fetuses, infants showed a discriminative neural response to syllables. Habituation was not observed in either fetuses or infants. These findings could be important for the investigation of early cognitive competencies and may help to gain a better understanding of language acquisition during child development.

In isolated tracheal smooth muscle preparations in normal rats and in rats with experimental fibrotic alveolitis, responses to electrical field stimulation of nervous and muscle fibers were studied. At stimulation of muscles or nerves of tracheal preparations without intramural ganglia in rats with acute alveolitis, parameters of smooth muscle contractions did not practically differ from those in normal rats. In rats with fibrotic alveolitis the amplitude and rate of muscle contraction decreased, while the response latent period (LP) increased. At stimulation of preganglionic nerve fibers of the tracheal preparations with intramural ganglia in rats with acute alveolitis, the value and rate of smooth muscle contraction decreased, while the response LP increased. After transition into chronic phase of the disease (fibrotic alveolitis), a partial restoration of the response parameters took place. In rats with acute alveolitis, the repeated stimulation of the nerve fibers led to an increase of amplitude and a decrease of rate of tracheal smooth muscle contractions. In rats with fibrotic alveolitis, the repeated stimulation caused a decrease of amplitude and rate of contractions and an increase of the response LP.

Changes in accommodative dynamics with repeated accommodation were studied in three anesthetized rhesus monkeys and two conscious humans. Maximum accommodation was centrally stimulated via the Edinger–Westphal nucleus in monkeys with a 4 s on, 4 s off paradigm (4 × 4) for 17 min, 4 × 1.5 for 27 min and 2 × 1 for 16 min. Humans accommodated repeatedly to visual targets (5 × 5; 5D and 2 × 2; 6D) for 30 min. In all cases, accommodation was sustained throughout. The anesthetized monkeys showed inter-individual variability in the extent of changes in accommodative dynamics over time while no systematic changes were detected in the human accommodative responses. Little accommodative fatigue was found compared to previous studies which have reported a complete loss of accommodation after 5 min of repeated stimulation in monkeys.

Intimidatory behaviour of nestling Bubo bengalensis to human intruders was investigated in five subjects. This disclosed a series of distinct behavioural unitsviz., hissing, ruffled plumage, bill clicking and the full intimidatory display - increasing in intensity from a crouched concealing posture and linked by short duration transition phases. Often no reaction to human presence was discernable. This was due to the effects of repeated stimulation/ of the parents’ alarm calls/ developmental stage of the young. The rationale for these intensity types and behavioural patterns, with emphasis on motivational and phylogenetic elements has been discussed.

Olfactory (H2S) and trigeminal (CO2) event-related potentials (ERPs) were studied with respect to circadian rhythm and desensitization. ERPs, perceived odor and pain intensity, oral temperature, blood pressure, heart rate, nasal volume, and sleepiness were assessed four times at 04:00, 08:00, 12:00, 16:00, 20:00, and 24:00 hr in five young men. For each of these 24 sessions per participant, H2S and CO2 were each presented in 15 series of five stimuli with a 5-s ISI within and 30-s ISI between series. ERP amplitudes, but not latencies, followed a circadian rhythm (largest at 16:00 and smallest at 04:00) similar to oral temperature and opposite to sleepiness. Amplitudes decreased (most pronounced at 16:00 and 20:00) and latencies increased with repeated stimulation, suggesting desensitization, in accordance with odor and pain intensity. These findings imply that circadian rhythm and desensitization should be considered in chemosensory ERP studies.

Many behavioural studies rely on playback experiments. While it is known that songbirds decrease behavioural responses after short-term repeated stimulation, long-term behavioural changes due to playbacks are unknown. We studied the response to playbacks in a free-living songbird in the long-term, while also studying the repeatability of the behaviour. Locomotor behaviour (a proxy of aggressiveness) decreased one year after first exposure to a single playback. Song response, however, remained consistent, suggesting that these two behaviours may provide different information. Locomotor behaviour was less repeatable than the song response to playback, the latter showing significant between-years repeatability. To the best of our knowledge, our study is the first to report long-term decrease in response to playbacks in a songbird, and that some aspects of the response to playback are repeatable. Similar studies in other species or populations of the great tit are important, to examine the generality of our findings.