Hypnotic Effects Research Articles

In Vitro and In Vivo Evaluation of Pellotine: A Hypnotic Lophophora Alkaloid.

Quality of life is often reduced in patients with sleep-wake disorders. Insomnia is commonly treated with benzodiazepines, despite their well-known side effects. Pellotine (1), a Lophophora alkaloid, has been reported to have short-acting sleep-inducing properties in humans. In this study, we set out to evaluate various in vitro and in vivo properties of 1. We demonstrate that 1 undergoes slow metabolism; e.g. in mouse liver microsomes 65% remained, and in human liver microsomes virtually no metabolism was observed after 4 h. In mouse liver microsomes, two phase I metabolites were identified: 7-desmethylpellotine and pellotine-N-oxide. In mice, the two diastereomers of pellotine-O-glucuronide were additionally identified as phase II metabolites. Furthermore, we demonstrated by DESI-MSI that 1 readily enters the central nervous system of rodents. Furthermore, radioligand-displacement assays showed that 1 is selective for the serotonergic system and in particular the serotonin (5-HT)1D, 5-HT6, and 5-HT7 receptors, where it binds with affinities in the nanomolar range (117, 170, and 394 nM, respectively). Additionally, 1 was functionally characterized at 5-HT6 and 5-HT7, where it was found to be an agonist at the former (EC50 = 94 nM, Emax = 32%) and an inverse agonist at the latter (EC50 = 291 nM, Emax = -98.6). Finally, we demonstrated that 1 dose-dependently decreases locomotion in mice, inhibits REM sleep, and promotes sleep fragmentation. Thus, we suggest that pellotine itself, and not an active metabolite, is responsible for the hypnotic effects and that these effects are possibly mediated through modulation of serotonergic receptors.

Detection of Flunitrazepam in Biological Fluids and Beverages:A Review

AbstractFlunitrazepam, a very potent benzodiazepine with sedative, hypnotic and amnesic effects, is commonly used to treat severe insomnia and as an anesthetic. Due to these characteristics, when flunitrazepam is sneaked into alcohol or drinks, it is often highly associated with violent behavior and drug rape cases. Therefore, it is also known as a “date rape drug”, and the detection of flunitrazepam and its metabolites is of great interest in forensic medicine and pharmacotoxicology. This paper describes the mechanism of action and drug metabolism process of flunitrazepam, reviews the different methods for detecting flunitrazepam published over the years, summarizes the detection limits of each method and compares their advantages and disadvantages. We hope to provide some inspiration for future research in this field.

Pharmacokinetics and Tissue Distribution of Isovitexin-2''-O-β-D-glucopyranoside (IVG) in Sprague-Dawley Rats.

Isovitexin-2"-O-D-glucopyranoside (IVG) has been known to exhibit sedative and hypnotic effects. However, there is little understanding of the in vivo pharmacokinetics and tissue distribution of IVG. This study aimed to investigate the pharmacokinetics and tissue distribution of IVG. The study employed an HPLC-ESI-MS/MS method to analyze the pharmacokinetics and tissue distribution of IVG. Under mass spectrometry, IVG and internal standard (IS) showed strong negative ionization signals. MRM analysis chose ion transitions m/z 593.3 → 293.0 (IVG) and m/z 579.8 → 271.4 (IS). Method validation indicated high precision, accuracy, and reliability with a quantitation limit under 20 ng/mL. After intravenously administering 5.0 mg/kg of IVG, rapid clearance from rat plasma was observed, with a half-life (t1/2) of 3.49 ± 0.99 h and a clearance rate of 54.53 ± 11.90 mL/kg/h. The area under the curve (AUC0-12h) of 37.79 ± 7.65 µg·h/mL indicated a brisk metabolic rate. Evaluating the tissue distribution, the highest accumulation was seen in the liver (30.32 ± 3.06 µg/g), followed by the kidney (20.58 ± 2.12 µg/g) and intestine (6.69 ± 0.93 µg/g), suggesting a propensity for IVG to concentrate in these tissues. Importantly, the presence of IVG in the brain underlines its potential to traverse the blood-brain barrier. These findings revealed that following intravenous administration, IVG was swiftly and broadly distributed throughout various rat tissues. This study provides valuable information on the pharmacokinetics and tissue distribution of IVG, implicating its potential as a novel and effective drug candidate for sedative and anxiolytic treatment.

Literature Review: Potensi Kombinasi Pijat dan Aromaterapi Lavender Terhadap Kualitas Tidur Bayi

Factors that affect the growth of the baby is the fulfillment of sleep needs, 75% of growth hormone is excreted when the baby sleeps, so it can stimulate brain development. However, babies often have difficulty sleeping due to various factors. Many studies have proven that massage using lavender aromatherapy has succeeded in improving the baby's sleep needs. Therefore, to summarize the results of this study, a literature study was carried out on the effect of lavender massage and aromatherapy. Literature review was carried out by searching scientific articles with keywords: aromatherapy-baby massage, aromatherapy massage on babies, baby massage and aromatherapy on babies. Article searches were performed on PubMed and Google Scholar. The search results obtained 7 articles that met the inclusion criteria. Based on the results of the literature review, it can be seen that the study population involved were infants aged 0-12 months. Baby massage combined with lavender aromatherapy can improve the quality of baby sleep and can affect the length of time to sleep and babies can easily fall back asleep when they wake up. Lavender aromatherapy can provide a calming effect, improve mood, blood circulation, maintain and increase endurance, sedative, hypnotic and anti-neurodepressive effects.

PHARMACOLOGICAL EVALUATION OF HYPNOTIC POTENTIAL OF BOMBAX CEIBA FLOWER EXTRACT IN RATS

Insomnia refers the problem with sleep depth or time duration and difficulties during initiation or maintaining sleep are symptoms- significantly distress and impair the daytime working. The Bombacaceae family, of which B. ceiba is a member, includes gigantic, beautiful deciduous trees native to Northern Australia. It can be found in both tropical and temperate regions of Africa, Australia, and Asia. So, the present current study was designed to find-out the hypnotic activity of flowers of B. ceiba and using animal model. The B. ceiba flowers were shipped in from the Northeast. A botanist confirmed its identity and legitimacy. The flowers are cleaned to remove any dust and dried in the shade or at room temperature. Albino rats (both sexes) weighing 130-150g were donated by the Animal House at the Institute of Pharmaceutical Sciences and Research, Unnao. Animals were taken as 6 in each group and treatments were done for 21 days I.E., Group 1 administered vehicle daily, Group 2 administered Diazepam (4mg/kg) orally, Group 3 administered ethanolic flower extracts of B. ceiba (EFBC) (200mg/kg) orally., Group 4 administered ethanolic flower extracts of B. ceiba (EFBC) (400mg/kg) orally. The hypnotic effect of above extract by following screening models i.e., Thiopental sod. induced sleeping time, Motor co-ordination, Light/ dark arena, Locomotion and Antioxidant activity. The study's findings showed that the flower extract of B. ceiba was very effective in treating insomnia and its symptoms. Its mode of action may involve either the downregulation of neurotransmitter release or the upregulation of catecholamine destruction. In concludes that flowers of Bombax ceiba are effective in relieving the symptoms of insomnia and improves status of mind in sleep induction. Its MOA is required to elaborate for its selection in curing different issues related to mental disorders. It also suggests the isolation of the significant active components of B. ceiba acting in activity and then converted into dosage form with facilitated absorption.

A Novel Probabilistic Diffusion Model Based on the Weak Selection Mimicry Theory for the Generation of Hypnotic Songs

The constraints in traditional music style transfer algorithms are difficult to control, thereby making it challenging to balance the diversity and quality of the generated music. This paper proposes a novel weak selection-based music generation algorithm that aims to enhance both the quality and the diversity of conditionally generated traditional diffusion model audio, and the proposed algorithm is applied to generate natural sleep music. In the inference generation process of natural sleep music, the evolutionary state is determined by evaluating the evolutionary factors in each iteration, while limiting the potential range of evolutionary rates of weak selection-based traits to increase the diversity of sleep music. Subjective and objective evaluation results reveal that the natural sleep music generated by the proposed algorithm has a more significant hypnotic effect than general sleep music and conforms to the rules of human hypnosis physiological characteristics.

Action of the general anaesthetic isoflurane reveals coupling between viscoelasticity and electrophysiological activity in individual neurons

General anaesthetics are widely used for their analgesic, immobilising, and hypnotic effects. The mechanisms underlying these effects remain unclear, but likely arise from alterations to cell microstructure, and potentially mechanics. Here we investigate this hypothesis using a custom experimental setup combining calcium imaging and nanoindentation to quantify the firing activity and mechanical properties of dorsal root ganglion-derived neurons exposed to a clinical concentration of 1% isoflurane gas, a halogenated ether commonly used in general anaesthesia. We found that cell viscoelasticity and functional activity are simultaneously and dynamically altered by isoflurane at different stages of exposure. Particularly, cell firing count correlated linearly with the neuronal loss tangent, the ratio of mechanical energy dissipation and storage by the cell. Our results demonstrate that anaesthetics affect cells as a whole, reconciling seemingly contradictory theories of how anaesthetics operate, and highlight the importance of considering cell mechanics in neuronal functions, anaesthesia, and clinical neuroscience in general.

Exopolysaccharide is the potential effector of Lactobacillus fermentum PS150, a hypnotic psychobiotic strain.

Psychobiotics are a class of probiotics that confer beneficial effects on the mental health of the host. We have previously reported hypnotic effects of a psychobiotic strain, Lactobacillus fermentum PS150 (PS150), which significantly shortens sleep latency in experimental mice, and effectively ameliorate sleep disturbances caused by either caffeine consumption or a novel environment. In the present study, we discovered a L. fermentum strain, GR1009, isolated from the same source of PS150, and found that GR1009 is phenotypically distinct but genetically similar to PS150. Compared with PS150, GR1009 have no significant hypnotic effects in the pentobarbital-induced sleep test in mice. In addition, we found that heat-killed PS150 exhibited hypnotic effects and altered the gut microbiota in a manner similar to live bacteria, suggesting that a heat-stable effector, such as exopolysaccharide (EPS), could be responsible for these effects. Our comparative genomics analysis also revealed distinct genetic characteristics in EPS biosynthesis between GR1009 and PS150. Furthermore, scanning electron microscopy imaging showed a sheet-like EPS structure in PS150, while GR1009 displayed no apparent EPS structure. Using the phenol-sulfate assay, we found that the sugar content value of the crude extract containing EPS (C-EPS) from PS150 was approximately five times higher than that of GR1009, indicating that GR1009 has a lower EPS production activity than PS150. Through the pentobarbital-induced sleep test, we confirmed the hypnotic effects of the C-EPS isolated from PS150, as evidenced by a significant reduction in sleep latency and recovery time following oral administration in mice. In summary, we utilized a comparative approach to delineate differences between PS150 and GR1009 and proposed that EPS may serve as a key factor that mediates the observed hypnotic effect.

The effect-site concentration of remifentanil blunting endotracheal intubation responses during anesthesia induction with etomidate: a dose-finding study

BackgroundRemifentanil can inhibit the hemodynamic responses caused by endotracheal intubation, but the effect-site concentration of it required to control intubation responses when combined with etomidate has not been demonstrated. The purpose of this study was to determine the effect-site concentration of remifentanil blunting tracheal intubation responses in 50% and 95% of patients (EC50 and EC95) during etomidate anesthesia.MethodsAmerican Society of Anesthesiologists physical status (ASA) I-II elective surgical patients receiving target-controlled infusion (TCI) of remifentanil, followed by etomidate and rocuronium for anesthesia were enrolled. The Belive Drive A2 monitor was used to calculate the MGRSSI (Maygreen Sedative state index) of hypnotic effect and the MGRNOX (Maygreen Nociception index) of nociception. The MGRSSI and the MGRNOX value were generated every 1 s. Mean arterial pressure (MAP) and heart rate (HR) were measured every minute, noninvasively. Using the modified Dixon’s up-and-down method, the concentration of remifentanil was determined based on the intubation response of the previous patient. The cardiovascular response during endotracheal intubation was defined as positive when MAP or HR is 20% higher than the pre-intubation value. A probit analysis was used for calculating EC50, EC95 and 95% confidence interval (CI).ResultsThe EC50 and EC95 of remifentanil blunting tracheal intubation responses were found to be 7.731 ng/ml (95%CI: 7.212–8.278 ng/ml) and 8.701 ng/ml (95%CI: 8.199–11.834 ng/ml). There were statistically significant increases in HR, MGRSSI and MGRNOX value to tracheal intubation in the positive responses group compared to the negative group. The most common adverse event was postoperative nausea and vomiting, which occurred in 3 patients.ConclusionRemifentanil effect-site concentration of 7.731 ng/ml is effective in blunting sympathetic responses to tracheal intubation in 50% of patients when combined with etomidate anesthesia.Trial registrationThe trial was registered at the Chinese Clinical Trials Registry (www.chictr.org.cn, registration number: ChiCTR2100054565, date of registration: 20/12/2021).

Activation of astrocytes in the basal forebrain in mice facilitates isoflurane-induced loss of consciousness and prolongs recovery

ObjectivesGeneral anesthesia results in a state of unconsciousness that is similar to sleep. In recent years, increasing evidence has reported that astrocytes play a crucial role in regulating sleep. However, whether astrocytes are involved in general anesthesia is unknown.MethodsIn the present study, the designer receptors exclusively activated by designer drugs (DREADDs) approach was utilized to specifically activate astrocytes in the basal forebrain (BF) and observed its effect on isoflurane anesthesia. One the other side, L-α-aminoadipic acid was used to selectively inhibit astrocytes in the BF and investigated its influence on isoflurane-induced hypnotic effect. During the anesthesia experiment, cortical electroencephalography (EEG) signals were recorded as well.ResultsThe chemogenetic activation group had a significantly shorter isoflurane induction time, longer recovery time, and higher delta power of EEG during anesthesia maintenance and recovery periods than the control group. Inhibition of astrocytes in the BF delayed isoflurane-induced loss of consciousness, promoted recovery, decreased delta power and increased beta and gamma power during maintenance and recovery periods.ConclusionsThe present study suggests that astrocytes in the BF region are involved in isoflurane anesthesia and may be a potential target for regulating the consciousness state of anesthesia.

Age-Related Effects of Exogenous Melatonin on Anxiety-like Behavior in C57/B6J Mice.

The synthesis of melatonin (MLT) physiologically decreases during aging. Treatment with MLT has shown anxiolytic, hypnotic, and analgesic effects, but little is known about possible age-dependent differences in its efficacy. Therefore, we studied the effects of MLT (20 mg/kg, intraperitoneal) on anxiety-like behavior (open field (OFT), elevated plus maze (EPMT), three-chamber sociability, and marble-burying (MBT) tests), and the medial prefrontal cortex (mPFC)-dorsal hippocampus (dHippo) circuit in adolescent (35-40 days old) and adult (three-five months old) C57BL/6 male mice. MLT did not show any effect in adolescents in the OFT and EPMT. In adults, compared to vehicles, it decreased locomotor activity and time spent in the center of the arena in the OFT and time spent in the open arms in the EPMT. In the MBT, no MLT effects were observed in both age groups. In the three-chamber sociability test, MLT decreased sociability and social novelty in adults, while it increased sociability in adolescents. Using local field potential recordings, we found higher mPFC-dHippo synchronization in the delta and low-theta frequency ranges in adults but not in adolescents after MLT treatment. Here, we show age-dependent differences in the effects of MLT in anxiety paradigms and in the modulation of the mPFC-dHippo circuit, indicating that when investigating the pharmacology of the MLT system, age can significantly impact the study outcomes.

Chaihu-Longgu-Muli Decoction promotes sleep in mice with insomnia

Purpose: To determine the hypnotic and sedative effects, and the mechanism of action of ChaihuLonggu-Muli Decoction (CLMD) in mice with insomnia.
 Methods: A mice model induced with insomnia using p-chlorophenylalanine (PCPA), was established. The sedative and hypnotic effects of CLMD on mice were evaluated by animal behavioral tests. The animals were separated into four experimental groups: (1) control group, (2) PCPA group, (3) PCPA + 5 mg/kg CLMD group, and (4) CPA + 10 mg/kg CLMD group. The 5-HTR1A expression was determined by western blot. The expression levels of the neurotransmitters were assessed using commercial ELISA kits, while the histological changes in hypothalamic tissues were examined microscopically after staining with hematoxylin and eosin (H & E).
 Results: CLMD exerted a sedative effect on and ameliorated anxiety-like and stressful behaviors in mice with insomnia. Moreover, CLMD induced sleep and enhanced 5-HTR1A to modulate neurotransmitter secretion. The data also indicate that CLMD exerted a protective role by alleviating hypothalamic damage and regulating the activation of NLRP3 inflammasome associated with insomnia.
 Conclusion: These findings indicate that CLMD has sedative and hypnotic effects, and offers a new therapeutic modality for insomnia treatment.

Hypnotic prescriptions in Japan may be shifting from benzodiazepine receptor agonists to other types of hypnotics, melatonin receptor agonists, and orexin receptor antagonists.

Hypnotic prescriptions in Japan may be shifting from benzodiazepine receptor agonists to other types of hypnotics, melatonin receptor agonists, and orexin receptor antagonists.

Research progress on processing historical evolution, chemical constituents, and pharmacological action of Bombyx Batryticatus

Bombyx Batryticatus is a precious traditional Chinese animal drug commonly used in clinical practice in China, which has the effects of extinguishing wind, stopping convulsions, dispelling wind, relieving pain, resolving phlegm, and dissipating mass. The processing of Bombyx Batryticatus has a long history. As early as in the Liu Song period of the Southern and Northern Dynasties, there was a record of the processing of Bombyx Batryticatus with rice swill. In addition to the processing with bran, honey bran, and ginger juice, which are still used today, there are also processing methods such as rendering, flour processing, wine processing, salt processing, oil processing, charcoal, and red dates processing in ancient times. After processing, the fishy smell of Bombyx Batryticatus can be removed, and avoid nausea and vomiting caused by the direct taking. Furthermore, processing can also facilitate the removal of surface hairs and toxicity reduction, making the medicinal material crispy and easy to crush. Previous studies have shown that the main chemical constituents of Bombyx Batryticatus include protein polypeptides, sterols, and flavonoids, with anticonvulsant, anticoagulation, antithrombotic, anti-cancer, hypnotic, hypoglycemic, and other pharmacological effects. This paper reviewed the processing historical evolution, chemical constituents, and pharmacological effects of Bombyx Batryticatus to lay a foundation for the research on the processing mechanism, quality control, and active core substances of Bombyx Batryticatus.

English

The objective of this study is to study the sedative and hypnotic effects of the compatibility of Schisandra chinensis (SC) and Polygala tenuifolia (PT) on insomnia mice and its mechanisms. The 160 SPF KM mice were divided into 8 groups with 20 each group (10 female and 10 male mice), including control, model, estazolam, SC, PT, SC-PT 1:1, SC-PT 1:2, and SC-PT 2:1 group. Insomnia mice model was established with intraperitoneal injection of P-chlorophenylalanine (PCPA) for 3 days. The mice were investigated by the sleep time of mice induced by pentobarbital sodium, the levels of GABA, DA, 5-HT and 5-HIAA in hypothalamus by enzyme-linked immunosorbent assay, and the GABAα1 and DRD2 mRNA expression in hypothalamus by quantitative real-time polymerase chain reaction (RT-qPCR). SC-PT reduced the sleep latency of insomnia mice and prolonged the Pentobarbital-sodium induced sleep time than model group. SC-PT 2:1 group increased the levels of 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA), γ-Aminobutyric acid (GABA) and the expression of GABAARα1 and decreased the levels of dopamine (DA) and the expression of DRD2 in the hypothalamus of insomnia mice. The compatibility of SC and PT can enhance the function of sedation and hypnosis than single group. This study provides a basis for the compatibility of SC and PT in the insomnia treatment. Key words: Schisandra chinensis, Polygala tenuifolia, mice, sedative, hypnotic, γ-Aminobutyric acid, dopamine.

Pharmacological Analysis of GABAA Receptor and Sigma1R Chaperone Interaction: Research Report I-Investigation of the Anxiolytic, Anticonvulsant and Hypnotic Effects of Allosteric GABAA Receptors' Ligands.

Two groups of facts have been established in previous drug development studies of the non-benzodiazepine anxiolytic fabomotizole. First, fabomotizole prevents stress-induced decrease in binding ability of the GABAA receptor's benzodiazepine site. Second, fabomotizole is a Sigma1R chaperone agonist, and exposure to Sigma1R antagonists blocks its anxiolytic effect. To prove our main hypothesis of Sigma1R involvement in GABAA receptor-dependent pharmacological effects, we performed a series of experiments on BALB/c and ICR mice using Sigma1R ligands to study anxiolytic effects of benzodiazepine tranquilizers diazepam (1 mg/kg i.p.) and phenazepam (0.1 mg/kg i.p.) in the elevated plus maze test, the anticonvulsant effects of diazepam (1 mg/kg i.p.) in the pentylenetetrazole-induced seizure model, and the hypnotic effects of pentobarbital (50 mg/kg i.p.). Sigma1R antagonists BD-1047 (1, 10, and 20 mg/kg i.p.), NE-100 (1 and 3 mg/kg i.p.), and Sigma1R agonist PRE-084 (1, 5, and 20 mg/kg i.p.) were used in the experiments. Sigma1R antagonists have been found to attenuate while Sigma1R agonists can enhance GABAARs-dependent pharmacological effects.

Evaluation and correlation of nociceptive response index and spectral entropy indices as monitors of nociception in anesthetized patients.

During anesthesia, the response to these stimuli depends on the balance between nociception and antinociception. Recently, various monitoring systems based on the variables derived from electroencephalography, plethysmography, autonomic tone, reflex pathways, and composite algorithms have been introduced for monitoring nociception. The main aim of our study was to evaluate and correlate the physiological variables which reflect the autonomic nervous system response to nociception, such as heart rate (HR), systolic blood pressure (SBP), perfusion index (PI), and nociceptive response index (NRI), with the spectral entropy indices response entropy (RE) and RE-state entropy (SE), which reflects electromyographic (EMG) activation as a response to pain. This is a retrospective analysis of the data from a prospective study on the hypnotic and analgesic effects and the recovery profile of sevoflurane-based general anesthesia. Eighty-six patients undergoing single-agent sevoflurane anesthesia were recruited in the study. The study parameters, HR, SBP, SE, RE, RE-SE, PI, and NRI, were recorded at predefined time points before and after a standardized noxious stimulus. Correlation between the variables was carried out by applying the Pearson correlation equation for normal and the Spearman correlation equation for non-normally distributed data. Receiver operating characteristic (ROC) graphs were plotted, and the area under the curve was calculated to assess the diagnostic accuracy of post-stimulus NRI in detecting pain which was defined as RE-SE >10. There was a significant increase in the SBP, HR, NRI, RE, SE, and RE-SE and a considerable decrease in PI values during the post-noxious period compared to the pre-noxious period. There was no correlation between the absolute values of NRI and entropy indices at T2. However, among the reaction values, there was a weak correlation between the reaction values of NRI and RE (r = 0.30; P = 0.05). The area under the ROC curve for NRI to detect pain as defined by RE-SE >10 was 0.56. During sevoflurane anesthesia, the application of noxious stimulus causes significant changes in variables reflecting sympathetic response and EMG activity. However, NRI failed to detect nociception, and there was only a weak correlation between the reaction values of NRI and RE-SE.

Amapola, lindísima amapola: de los opiáceos y los opioides, utilidad y riesgos.

Opium and its derivatives, and recently the opioids have ac companied the humankind since the ancient civilizations to the present day. Its analgesic, hypnotic and pleasant effects did not go unnoticed by ancient people, which considered most of these effects of medical utility and noticed that they had remarkable mood benefits. Currently, there are no other kind of drugs that can palliate intense pain as efficiently as these powerful analgesics. However, the medical and rec reational use of opiates and opioids may carry health risks such as tolerance, hyperalgesia, and addiction. Nowadays, in addition to being indisputably the most powerful medical treatment to alleviate the suffering caused by pain, it has also become a public health problem due to the high number of people with opioid use disorder that have facilitated deaths caused by opioids overdose. In this review we will discuss the medical benefits of opiates and opioids, as much as the unwanted effects they produce. Keywords: Opioids; opiates; medical utility; health risks; addiction; abuse.

Heukharang lettuce (Lactuca sativa L.) leaf extract displays sleep-promoting effects through GABAA receptor

Ethnopharmacological relevanceAlthough lettuce is traditionally known to have hypnotic and sedative effects, to date, only a few studies have documented its sleep-promoting effects and elucidated the related mechanisms. Aim of the studyWe aimed to investigate the sleep-promoting activity of Heukharang lettuce leaf extract (HLE) with increased lactucin content, known as a sleep-promoting substance in lettuce, in animal models. Materials and methodsTo evaluate the effect of HLE on sleep behavior, analysis of electroencephalogram (EEG), gene expression of brain receptors, and activation mechanisms using antagonists were investigated in rodent models. ResultsHigh-performance liquid chromatography analysis showed that HLE contained lactucin (0.78 mg/g of extract) and quercetin-3-glucuronide (1.3 mg/g of extract). In the pentobarbital-induced sleep model, the group administered 150 mg/kg of HLE showed a 47.3% increase in sleep duration time as compared to the normal group (NOR). The EEG analysis showed that the HLE significantly increased non-rapid eye movement (NREM), where delta waves were improved by 59.5% when compared to the NOR, resulting in increased sleep time. In the caffeine-induced arousal model, HLE significantly decreased the awake time increased by caffeine administration (35.5%) and showed a similar level to NOR. In addition, HLE increased the gene and protein expression of gamma-aminobutyric acid receptor type A (GABAA), GABA type B, and 5-hydroxytryptamine (serotonin) receptor 1A. In particular, in comparison to the NOR, the group administered 150 mg/kg HLE showed an increase in expression levels of GABAA and protein by 2.3 and 2.5 times, respectively. When the expression levels were checked using GABAA receptor antagonists, HLE showed similar levels to NOR, as the sleep duration was reduced by flumazenil (45.1%), a benzodiazepine antagonist. ConclusionsHLE increased NREM sleep and significantly improved sleep behavior due to its action on the GABAA receptors. The collective findings suggest that HLE can be used as a novel sleep-enhancing agent in the pharmaceutical and food industries.

24 Effects of Oral Supplementation of Cannabidiol on Young Stallion Behavior

Abstract Cannabidiol (CBD) has become a popular product in the recent months due to change in legislation with the passing of the Agricultural Improvement Act in 2018. Cannabidiol comes from the plant Cannabis sativa L. and binds to cannabinoid receptors throughout the endocannabinoid system. These receptors are located throughout the body and are prominent in the brain and reproduction tract. Although the effects of CBD on physiology have been researched, little data on the behavioral effects exist, even less so in horses. Calming supplements that may negate aggressive stallion behavior are not suitable due to potential soporific effects that can affect libido. In addition to the effectiveness of CBD to modify behavior in horses, it is also unclear if CBD influences stallion libido. The objective of this study was to evaluate sexual behavior in stallions given a daily CBD supplement for 70 days. Two-year old stallions (n = 8) were assigned to either a Control (CON; n = 3) or Cannabidiol (CBD; n = 4) treatment groups. Cannabidiol was administered at a rate of 0.6 mg/kg BW daily, and equal volume of olive oil was given as a placebo to CON. Horses were weighed every Monday at which time dosages were recalculated to adjust for body weight changes. Behavior data collection occurred twice per week during semen collections. Video data were recorded by placing two cameras, one placed in view of the tease area, and one placed in view of the mount, for behavior data backup. Behavioral data was tallied by handlers calmly calling each behavior as it occurred during collection. The behaviors collected included Flehman’s response, bites, strikes, mounts, high pitch vocalizations, and low pitch vocalizations. Data were analyzed using a Chi Square Test within R Statistical Program (R Core Team). Results were considered significant at P < 0.05. Throughout the duration of the study, all stallions exhibited normal sexual behavior, regardless of treatment. Results showed that total low and high pitch vocalizations, bites, and Flehman’s were affected by treatment (P < 0.01). Cannabidiol-treated stallions had more low pitches (60), compared to CON (58). Cannabidiol-treated stallions had fewer bites (145) compared to the CON (146). Cannibidiol stallions displayed Flehman’s more frequently (270) than CON (236). Flehman’s response and bites were lower for stallions on cannabidiol in Week 1 but increased by Week 10 (P < 0.01), while only Flehman’s response increased in the CON group over time (P < 0.01). These findings indicate that CBD may influence breeding behavior in young stallions specifically when supplemented over time. Further research should focus on the impact of CBD on breeding behaviors in mature stallions and the longevity of the behavioral response to CBD.