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Goal: We validate a recent reverse correlation approach to tinnitus characterization by applying it to individuals with clinically-diagnosed tinnitus. Methods: Two tinnitus patients assessed the subjective similarity of their non-tonal tinnitus percepts and random auditory stimuli. Regression of the responses onto the stimuli yielded reconstructions which were evaluated qualitatively by playing back resynthesized waveforms to the subjects and quantitatively by response prediction analysis. Results: Subject 1 preferred their resynthesis to white noise; subject 2 did not. Response prediction balanced accuracies were significantly higher than chance across subjects: subject 1: 0.5963, subject 2: 0.6922. Conclusion: Reverse correlation can provide the foundation for reconstructing accurate representations of complex, non-tonal tinnitus in clinically diagnosed subjects. Further refinements may yield highly similar waveforms to individualized tinnitus percepts.

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Glyphodes pyloalis Walker (Lepidoptera: Pyralidae) is the most destructive pest, causing severe damage to mulberry production in China's sericulture industry. The insecticide application in mulberry orchards poses a significant risk of poisoning to Bombyx mori. Shifting from insecticides to odor attractants is a beneficial alternative, but not much data is available on the olfactory system of G. pyloalis. We identified 114 chemosensory genes from the antennal transcriptome database of G. pyloalis, with 18 odorant-binding protein (OBP) and 17 chemosensory protein (CSP) genes significantly expressed in the antennae. Ligand-binding assays for two antennae-biased expressed general odorant-binding proteins (GOBPs) showed high binding affinities of GOBP1 to hexadecanal, ß-ionone, and 2-ethylhexyl acrylate, while GOBP2 exhibited binding to 4-tert-octylphenol, benzyl benzoate, ß-ionone, and farnesol. Computational simulations indicated that van der Waal forces predominantly contributed to the binding free energy in the binding processes of complexes. Among them, Phe12 of GOBP1 and Phe19 of GOBP2 were demonstrated to play crucial roles in their bindings to plant volatiles using site-directed mutagenesis experiments. Moreover, hexadecanal and ß-ionone attracted G. pyloalis male moths in the behavioral assays, while none of the candidate plant volatiles significantly affected female moths. Our findings provide a comprehensive understanding of the molecular mechanisms underlying olfactory recognition in G. pyloalis, setting the groundwork for novel mulberry pests control strategies based on insect olfaction.

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Aethina tumida (small hive beetle, SHB) is a rapidly spreading invasive parasite of bee colonies. The olfactory system plays a key role in insect behavior, and odorant-binding proteins (OBPs) are involved in the first step of the olfactory signal transduction pathway and the detection of host volatiles. However, the olfactory mechanism of OBPs in SHB-localized bee colonies is unclear. In this study, electroantennogram (EAG) and behavioral bioassay showed that only three compounds (2-heptanone, ocimene, and ethyl palmitate) from bee colonies triggered high electrophysiological and behavioral responses. Three antenna-specific OBP genes (OBP6, OBP11, and OBP19) were identified, and they were significantly expressed on adult days 6-7. Furthermore, by combining RNA interference (RNAi) with EAG, olfactometer bioassay, competitive fluorescence binding assays, and molecular docking, we found that these three OBP genes were involved in the recognition of 2-heptanone and ethyl palmitate, and AtumOBP6 is also involved in the recognition of ocimene. These data indicate that AtumOBP6, AtumOBP11, and AtumOBP19 play an important role in the olfactory response to bee colony volatiles. Our results provide new insights into the functions of the OBP families in A. tumida and help to explore more potential target genes for environmentally friendly pest control strategies.

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For aquatic ecosystem Daphnia magna is evolving as a model organism to check the teratogenicity of numerous compounds. D. magna can be easily cultured in the laboratory, and the teratogen effect of several compounds can be easily studied. The developmental stages are well studied in D. magna. All the developmental stages are transparent so the defect can be easily accessed. So, the postembryonic developmental changes can be easily studied after the exposure with teratogen. More importantly, D. magna also have a swimming behavioral phenotype. The behavioral defect can be easily accessed after teratogen exposure. The current chapter summarizes numerous protocols associated with embryo and adult staining and adult behavioral assays that can be used to access the teratogenicity of any unknown compound.

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The fruit fly Drosophila melanogaster is a powerful genetic model that has been used for many decades to study nervous system function, development, and behavior. There are a large number of developmental and behavioral traits that can be measured to provide a broad readout of neurological function. These include patterned motor behaviors, such as larval locomotion, which can be used to assess whether genetic or environmental factors affect nervous system function to provide an entry point for deeper mechanistic studies. Here, we describe a protocol for quantifying larval locomotion using a simple camera setup and a freely available image analysis software. This protocol can be readily applied to human disease models or in toxicology studies, for example, to broadly assess the impact of treatments on neurological function.

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Sex pheromones are considered to play critical roles in partner communication of most parasitic Hymenoptera. However, the identification of sex pheromone components remains limited to a few families of parasitoid wasps. In this study, we functionally characterized a candidate sex pheromone component in Microplitis mediator (Hymenoptera: Braconidae), a solitary parasitoid of Noctuidae insects. We found that the body surface extract from female wasps could significantly stimulate courtship behavior of males. Gas chromatography-electroantennographic detection (GC-EAD) analysis revealed that a candidate semiochemical from extract triggered significant electrophysiological response of antennae of males. By performing gas chromatography-mass spectrometer (GC-MS) measurement, GC-EAD active compound was identified as n-octyl acrylate, a candidate sex pheromone component in female M. mediator. In electroantennogram (EAG) tests, antennae of male wasps showed significantly higher electrophysiological responses to n-octyl acrylate than those of females. Y-tube olfactometer assays indicated that male wasps significantly chose n-octyl acrylate compared with the control. Furthermore, male wasps showed a remarkable preference for n-octyl acrylate in a simulated field condition behavioral trial; simultaneously, n-octyl acrylate standard could also trigger significant courtship behavior in males. We propose that n-octyl acrylate, as a candidate vital sex pheromone component, could be utilized to design behavioral regulators of M. mediator to implement the protection and utilization of natural enemies.

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Eugenol, the major active ingredient of clove oil, is widely used for anesthesia in fish. Yet virtually nothing is known about its effects on CNS functions, and thus about potential interference with neurophysiological experimentation. To address this issue, we employed a neuro-behavioral assay recently developed for testing of water-soluble anesthetic agents. The unique feature of this in-vivo tool is that it utilizes a readily accessible behavior, the electric organ discharge (EOD), as a proxy of the neural activity generated by a brainstem oscillator, the pacemaker nucleus, in the weakly electric fish Apteronotus leptorhynchus. A deep state of anesthesia, as assessed by the cessation of locomotor activity, was induced within less than 3 min at concentrations of 30-60 µL/L eugenol. This change in locomotor activity was paralleled by a dose-dependent, pronounced decrease in EOD frequency. After removal of the fish from the anesthetic solution, the frequency returned to baseline levels within 30 min. Eugenol also led to a significant increase in the rate of 'chirps,' specific amplitude/frequency modulations of the EOD, during the 30 min after the fish's exposure to the anesthetic. At 60 µL/L, eugenol induced a collapse of the EOD amplitude after about 3.5 min in half of the fish tested. The results of our study indicate strong effects of eugenol on CNS functions. We hypothesize that these effects are mediated by the established pharmacological activity of eugenol to block the generation of action potentials and to reduce the excitability of neurons; as well as to potentiate GABAA-receptor responses.

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Social motivation is critical to the development of typical social functioning. Social motivation, specifically one or more of its components (e.g., social reward seeking or social orienting), could be relevant for understanding phenotypes related to autism. We developed a social operant conditioning task to quantify effort to access a social partner and concurrent social orienting in mice. We established that mice will work for access to a social partner, identified sex differences, and observed high test-retest reliability. We then benchmarked the method with two test-case manipulations. Shank3B mutants exhibited reduced social orienting and failed to show social reward seeking. Oxytocin receptor antagonism decreased social motivation, consistent with its role in social reward circuitry. Overall, we believe that this method provides a valuable addition to the assessment of social phenotypes in rodent models of autism and the mapping of potentially sex-specific social motivation neural circuits.

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BACKGROUND: Rhynchophorus ferrugineus, the red palm weevil (RPW), is a key pest that attacks many economically important palm species and that has evolved a sensitive and specific olfactory system to seek palm hosts. Odorant-binding proteins (OBPs) not only play crucial roles in its olfactory perception process but are also important molecular targets for the development of new approaches for pest management. RESULTS: Analysis of the tissue expression profiles of RferOBP8 and RferOBP11 revealed that these two Rhynchophorus ferrugineus odorant binding proteins (RferOBPs) exhibited high expression in the antennae and showed sexual dimorphism. We analyzed the volatiles of seven host plants by gas chromatography-mass spectrometry and screened 13 potential ligands by molecular docking. The binding affinity of two recombinant OBPs to aggregation pheromones and 13 palm odorants was tested by fluorescence competitive binding assays. The results revealed that eight tested palm volatiles and ferrugineol have high binding affinities with RferOBP8 or RferOBP11. Behavioral trials showed that these eight odor compounds could elicit an attraction response in adult RPW. RNA interference analysis indicated that the reduction in the expression levels of the two RferOBPs led to a decrease in behavioral responses to these volatiles. CONCLUSION: These results suggest that RferOBP8 and RferOBP11 are involved in mediating the responses of RPW to palm volatiles and to aggregation pheromones and may play important roles in RPW host-seeking. This study also provides a theoretical foundation for the promising application of novel molecular targets in the development of new behavioral interference strategies for RPW management in the future. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Goal: This study validates an approach to characterizing the sounds experienced by tinnitus patients via reverse correlation, with potential for characterizing a wider range of sounds than currently possible. Methods: Ten normal-hearing subjects assessed the subjective similarity of random auditory stimuli and target tinnitus-like sounds ("buzzing" and "roaring"). Reconstructions of the targets were obtained by regressing subject responses on the stimuli, and were compared for accuracy to the frequency spectra of the targets using Pearson's [Formula: see text]. Results: Reconstruction accuracy was significantly higher than chance across subjects: buzzing: [Formula: see text] (mean [Formula: see text] s.d.), [Formula: see text], [Formula: see text]; roaring: [Formula: see text], [Formula: see text], [Formula: see text]; combined: [Formula: see text], [Formula: see text], [Formula: see text]. Conclusion: Reverse correlation can accurately reconstruct non-tonal tinnitus-like sounds in normal-hearing subjects, indicating its potential for characterizing the sounds experienced by patients with non-tonal tinnitus.

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The growing consumption of antidepressant pharmaceuticals has resulted in their widespread occurrence in the environment, particularly in waterways with a typical concentration range from ng L-1 to µg L-1. An increasing number of studies have confirmed the ecotoxic potency of antidepressants, not only at high concentrations but also at environmentally relevant levels. The present review covers literature from the last decade on the individual-level ecotoxicological effects of the most commonly used antidepressants, including their impact on behavior, growth, and survival. We focus on the relationship between antidepressants physico-chemical properties and dynamics in the environment. Furthermore, we discuss the advantages of considering behavioral changes as sensitive endpoints in ecotoxicology, as well as some current methodological shortcomings in the field, including low standardization, reproducibility and context-dependency.

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The limited axon regeneration capacity of mature neurons often leads to insufficient functional recovery after damage to the central nervous system (CNS). To promote CNS nerve repair, there is an urgent need to understand the regeneration machinery in order to develop effective clinical therapies. To this aim, we developed a Drosophila sensory neuron injury model and the accompanying behavioral assay to examine axon regeneration competence and functional recovery after injury in the peripheral and central nervous systems. Specifically, we used a two-photon laser to induce axotomy and performed live imaging to assess axon regeneration, combined with the analysis of the thermonociceptive behavior as a readout of functional recovery. Using this model, we found that the RNA 3'-terminal phosphate cyclase (Rtca), which acts as a regulator for RNA repair and splicing, responds to injury-induced cellular stress and impedes axon regeneration after axon breakage. Here we describe how we utilize our Drosophila model to assess the role of Rtca during neuroregeneration.

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Aim: Cancer patients undergoing chemotherapy report cognitive changes collectively termed "chemo brain." Neuroinflammation is among the factors believed to contribute to "chemo brain" suggesting a potential beneficial role for anti-inflammatory drugs in cancer patients undergoing chemotherapy. We investigated whether the non-steroidal anti-inflammatory drug naproxen influenced hippocampal inflammation in non-tumor bearing female mice receiving the chemotherapy drug cyclophosphamide (CP).Materials and methods: Intact and ovariectomized C57BL/6 mice were used to examine potential role of ovarian hormones on neuroinflammation. The mice were placed on naproxen (375 ppm) or control diet, and a week later CP (100 mg/kg; i.p.) was administered every 3 days for 2 weeks. We analyzed hippocampal inflammatory biomarkers, anxiety-like behavior, spatial working memory, exploratory behavior, spontaneous locomotor activity and depression-like behavior.Results: CP produced significant effects on anti-inflammatory but not pro-inflammatory biomarkers. However, CP and naproxen in combination produced significant effects on both pro- and anti- inflammatory biomarkers. Naproxen and ovariectomy individually produced significant effects on pro- and anti-inflammatory biomarkers as well. Working memory and depression-like behavior were not significantly influenced by CP, naproxen or ovariectomy individually although CP and ovariectomy produced significant interaction effects on depression-like behavior. Exploratory behavior and locomotor activity showed significant effects of CP, and interaction between CP and naproxen was significant for locomotor activity.Conclusions: Ovariectomy, naproxen and a combination of CP and naproxen upregulate hippocampal pro- and anti- inflammatory biomarkers. None of the factors individually produce significant behavioral changes that could be consistent with chemo brain, although CP and ovariectomy in combination produced significant effects on depression-like behavior, a co-morbidity of chemo brain.

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Adaptation in the sensory-mechanical loop during locomotion is a powerful mechanism that allows organisms to survive in different conditions and environments. Motile animals need to alter motion patterns in different environments. For example, crocodiles and other animals can walk on solid ground but switch to swimming in water beds. The nematode Caenorhabditis elegans also shows adaptability by employing thrashing behaviour in low viscosity media and crawling in high viscosity media. The mechanism that enables this adaptability is an active area of research. It has been attributed previously to neuro-modulation by dopamine and serotonin. This study introduces an experimental assay to physiologically investigate the neuronal mechanisms of modulation of locomotion by dopamine. The technique is utilized to test gait switching while imaging the mechanosensory dopaminergic neurons PDE. Results revealed their role to be not limited to touch sensation, but to sensing surrounding environment resistance as well. The significance of such characterization is improving our understanding of dopamine gait switching which gets impaired in Parkinson's disease.-A locomotion pattern switching system was devised to allow studying this process in vivo in the nematode C. elegans.-This system allowed the study of dopaminergic neurons PDE response as the worms switched from crawling to swimming.

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C. elegans shows robust and reproducible behavioral responses to oxygen. Specifically, worms prefer O 2 levels of 5-10% and avoid too high or too low O 2 . Their O 2 preference is not fixed but shows plasticity depending on experience, context, or genetic background. We recently showed that this experience-dependent plasticity declines with age, providing a useful behavioral readout for studying the mechanisms of age-related decline of neural plasticity. Here, we describe a technique to visualize behavioral O 2 preference and its plasticity in C. elegans , by creating spatial gradients of [O 2 ] in a microfluidic polydimethylsiloxane (PDMS) chamber and recording the resulting spatial distribution of the animals.

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[This corrects the article DOI: 10.3389/fphys.2022.854124.].

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The granary weevil Sitophilus granarius (L.) is one of the most serious primary insect pests of stored products. When S. granarius present in grains, various volatile organic compounds are released as chemical signals which can be used to detect the insects. In this study, volatile chemical compounds released from S. granarius were analyzed using the headspace solid phase micro-extraction (HS-SPME) coupled with gas chromatography (GC)-mass spectrometry (MS) techniques. Two key compounds, 3-hydroxy-2-butanone and 1-pentadecene, were identified from mixed gender of S. granarius adults at high density. Moreover, both male and female adults showed dose-dependent electroantennography (EAG) responses to 3-hydroxy-2-butanone. In behavioral assays, S. granarius was attracted to 3-hydroxy-2-butanone at 0.001 µg/10 µL but repelled at 10 µg/10 µL or higher. S. granarius was consistently repelled by 1-pentadecene at concentrations at 0.001 and 1000 µg/10 µL. 3-hydroxy-2-butanone and 1-pentadecene have considerable potential to offer in the development of new approaches for the monitoring and management of this destructive stored grain insect pest.

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Nociception refers to the process of encoding and processing noxious stimuli, which allow animals to detect and avoid potentially harmful stimuli. Several types of stimuli can trigger nociceptive sensory transduction, including thermal, noxious chemicals, and harsh mechanical stimulation that depend on the corresponding nociceptors. In view of the high evolutionary conservation of the mechanisms that govern nociception from Drosophila melanogaster to mammals, investigation in the fruit fly Drosophila help us understand how the sensory nervous system works and what happen in nociception. Here, we present an overview of currently identified conserved genetics of nociception, the nociceptive sensory neurons responsible for detecting noxious stimuli, and various assays for evaluating different nociception. Finally, we cover development of anti-pain drug using fly model. These comparisons illustrate the value of using Drosophila as model for uncovering nociception mechanisms, which are essential for identifying new treatment goals and developing novel analgesics that are applicable to human health.

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Hydrothermal carbonization is considered a powerful technology to convert sewage sludge (SS) into a valuable carbonaceous solid known as hydrochar (HC). Up to now criteria for landfill application of SS and HC are based only on physicochemical properties and levels of pollutant residues. Nevertheless, to ensure their safe environmental applications it is mandatory to develop biosensors which can provide relevant information on their toxic potential for natural ecosystems. Therefore, this study aimed to assess the suitability of a contact assay using zebrafish embryo/larvae combined with sub-lethal end-points to evaluate the hazard associated with SS and related HC exposure. A suite of biomarkers was also applied on larvae, related to detoxification and oxidative stress as the activity of Ethoxyresorufin-O-deethylase, glutathione-S-transferase, and catalase, the content of reactive oxygen species and the behavioral assay using the DanioVision™ chamber. Legacy priority pollutants were also measured either in SS and HC tested samples and in contact waters. The exposure to SS caused higher lethality compared to HC. No significant changes in the activity of oxidative stress markers was observed upon exposure to both matrices. The behavioral test showed a hypoactivity condition in larvae exposed to both SS and HC with the effects of SS stronger than HC. Chemical analysis revealed the presence of trace elements and halogenated compounds in either SS, HC. Heavy metals were also released in contact waters, while volatile hydrocarbons (C6-C10) and halogenated compounds resulted below LOD (<0.05 µ L-1). Our study highlights the suitability of zebrafish embryotoxicity test, coupled with behavioral traits, as screening tool for assessing potential risks, associated with the landfill application of both SS and HC, for aquatic wildlife.

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The photomotor response (PMR) of zebrafish embryos, a light pulse-triggered undirected movement, is known to be altered by neuroactive chemicals. Here, we developed an approach for data analysis of the distribution of PMR movement activities along the time axis; differences between treatment and respective controls are expressed by an aggregated value integrating the time-resolved density of the movement parameter as a measure for a chemically elicited PMR effect. Logistic concentration-PMR effect relationships were modeled for neuroactive test compounds with different modes of action (acetylcholinesterase inhibition, activation and inhibition of voltage-gated sodium channels); 50% effect concentrations (EC50) were in the low to medium µM range (EC50 < 10 µM for flucythrinate, esfenvalerate, azinphos-methyl, propoxur; EC50 > 10 µM for tricaine). Modulation of movement activities in different phases of the PMR (i.e., "fingerprint") by neuroactive test compounds varied across concentrations, showing that mode of action-specific PMR fingerprints are also concentration-dependent. Above concentrations causing 10% lethality (LC10; 48 h), 3,4-dichlo­roaniline caused movement inhibition. This substance presumably is not neuroactive; its effect on the PMR therefore is considered a secondary toxic effect. Quantitative morphological examinations of chemically exposed embryos showed that malformations occurred only above PMR effect concentrations, indicating that changes in the PMR were not due to such indirect effects. The PMR assay will provide a useful measure in ecotoxicological risk assessment of neuroactive chemicals with zebrafish embryos and could potentially be used to infer acute fish toxicity levels from PMR effect concen­trations of neurotoxic compounds.

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