Protection by Nano-Encapsulated Bacoside A and Bacopaside I in Seizure Alleviation and Improvement in Sleep- In Vitro and In Vivo Evidences.

Therapeutic options to contain seizures, a transitional stage of many neuropathologies, are limited due to the blood-brain barrier (BBB). Herbal nanoparticle formulations can be employed to enhance seizure prognosis. Bacoside A (BM3) and bacopaside I (BM4) were isolated from Bacopa monnieri and synthesized as nanoparticles (BM3NP and BM4NP, respectively) for an effective delivery system to alleviate seizures and associated conditions. After physicochemical characterization, cell viability was assessed on mouse neuronal stem cells (mNSC) and neuroblastoma cells (N2a). Thereafter, anti-seizure effects, mitochondrial membrane potential (MMP), apoptosis, immunostaining and epileptic marker mRNA expression were determined in vitro. The seizure-induced changes in the cortical electroencephalogram (EEG), electromyography (EMG), Non-Rapid Eye Movement (NREM) and Rapid Eye Movement (REM) sleep were monitored in vivo in a kainic acid (KA)-induced rat seizure model. The sizes of BM3NPs and BM4NPs were 165.5 nm and 689.6 nm, respectively. They were biocompatible and also aided in neuroplasticity in mNSC. BM3NPs and BM4NPs depicted more than 50% cell viability in N2a cells, with IC50 values of 1609 and 2962 µg/mL, respectively. Similarly, these nanoparticles reduced the cytotoxicity of N2a cells upon KA treatment. Nanoparticles decreased the expression of epileptic markers like fractalkine, HMGB1, FOXO3a and pro-inflammatory cytokines (P < 0.05). They protected neurons from apoptosis and restored MMP. After administration of BM3NPs and BM4NPs, KA-treated rats attained a significant reduction in the epileptic spikes, sleep latency and an increase in NREM sleep duration. Results indicate the potential of BM3NPs and BM4NPs in neutralizing the KA-induced excitotoxic seizures in neurons.

Bifunctional cysteine gold nanoclusters for ß-amyloid fibril inhibition and fluorescence imaging: a distinctive approach to manage Alzheimer's disease.

Alzheimer's disease (AD) is a progressive complex neurodegenerative disorder affecting millions of individuals worldwide. Currently, there is no effective treatment for AD. AD is characterized by the deposition of amyloid plaques/fibrils. One major strategy for managing this disease is by slowing the progression of AD using different drugs which could potentially limit free-radical formation, oxidative stress and lipid peroxidation and promote the survival of neurons exposed to ß-amyloid. Inhibition of amyloid fibrillization and clearance of amyloid plaques/fibrils are essential for the prevention and treatment of AD. The thiophilic interaction between the side chain of an aromatic residue in a polypeptide and a sulphur atom of the compound can effectively inhibit amyloid fibril formation. In this work, we have synthesized cysteine-capped gold nanoclusters (Cy-AuNCs) which exhibit inherent red emission and can disintegrate amyloid fibrils through the aforementioned thiophilic interactions. Herein, we also used molecular docking to study the thiophilic interactions between the sulphur atom of Cy-AuNCs and the aromatic rings of the protein. Finally, the gold cluster was functionalized with a brain targeting molecule, Levodopa (AuCs-LD), to specifically target the brain and to facilitate passage through the blood brain barrier (BBB). Both Cy-AuNCs and AuCs-LD showed good biocompatibility and the inherent fluorescence properties of nanoclusters enabled real time imaging. The efficacy of the nanoclusters to disintegrate amyloid fibrils and their ability to cross the BBB were demonstrated both in vitro and in vivo in the BBB model and the AD animal model respectively. Our results imply that nanoparticle-based artificial molecular chaperones may offer a promising therapeutic approach for AD.

Yoga Nidra, a Nonpharmacological Technique in Management of Insomnia and Overall Health in Postmenopausal Women.

Yoga Nidra is a promising technique through which body is consciously simulated into a profound relaxation state similar to attained during naturally occurring deep sleep. It is aimed to attain complete emotional, physical, and mental relaxation of body and mind. In postmenopausal phase of life, regular practice of Yoga Nidra at home preferably in morning, can help in reduction in anxiety and pain associated with early morning awakenings. This nonpharmacological technique has a therapeutic potential to improve sleep quality and quantity, and overall well-being.

Yogic Sleep and Walking Protocol Induced Improvement in Sleep and Wellbeing in Post-menopausal Subject: A Longitudinal Case Study During COVID Lockdown.

Purpose: Post-menopausal life is fairly long period of life that is marked by poor health and sleep. Fatigue amidst extraordinary pandemic stress had taken a toll on the sleep quality and overall wellbeing. Yogic sleep can be instrumental in relaxing the brain and help in achieving self-control of mind and body in the post-menopausal life. This can be a non-pharmacological intervention to improve the wellbeing of women. Methods: Effect of 24 weeks of yoga-nidra practice and exercise module was tested in a post-menopausal subject after taking baseline of 4 weeks on parameters like sleep latency, total sleep time, mood on waking and during day, BMI, and activity rhythm of body using 24 h actigraphy and sleep diary. Results: After administering the dual protocol, there was remarkable elevation in mood both on waking up and entire day from 5th week onwards. Mood shifted toward a happier state. Latency to sleep decreased after 4 weeks, while total sleep time improved only after 16 weeks of dual management strategy. The BMI was also reduced to 28.4 from initial value of 30.3. Morning awakening patterns did not change, but it was not accompanied by pain or headache. Conclusion: The results indicated the therapeutic potential of yoga-nidra and exercise package in this actigraphy-based longitudinal pilot study. Yoga-nidra can be easily practiced at home, and thus, it is a promising non-pharmacological strategy for aging population in improving their wellbeing.

Effects of sleep modulation during pregnancy in the mother and offspring: Evidences from preclinical research.

Disturbed sleep during gestation may lead to adverse outcomes for both mother and child. Animal research plays an important role in providing insights into this research field by enabling ethical and methodological requirements that are not possible in humans. Here, we present an overview and discuss the main research findings related to the effects of prenatal sleep deprivation in animal models. Using systematic review approaches, we retrieved 42 articles dealing with some type of sleep alteration. The most frequent research topics in this context were maternal sleep deprivation, maternal behaviour, offspring behaviour, development of sleep-wake cycles in the offspring, hippocampal neurodevelopment, pregnancy viability, renal physiology, hypertension and metabolism. This overview indicates that the number of basic studies in this field is growing, and provides biological plausibility to suggest that sleep disturbances might be detrimental to both mother and offspring by promoting increased risk at the behavioural, hormonal, electrophysiological, metabolic and epigenetic levels. More studies on the effects of maternal sleep deprivation are needed, in light of their major translational perspective.

Immature sleep pattern in newborn rats when dams encountered sleep restriction during pregnancy.

There is a growing realization that proper sleep during pregnancy is essential for the health of the mother and the offspring. However, there are no reports on the effects of maternal sleep restriction on the sleep-wake profiles of newborns. So, this study was conducted to evaluate the effects of sleep restriction during the third term of pregnancy on the sleep-wake profiles of neonates born to them. The female pregnant Wistar rats were sleep restricted for 5 h/day on gestational days 15-20 by gentle handling. Sleep-wake profiles of the pups born to them and to the control rats were recorded on postnatal days 1-21. Pups of sleep restricted dams had higher active sleep (AS) and lower quiet sleep (QS) as well as wakefulness. Higher ratio of AS to QS, longer duration of sleep cycles, lesser bout frequency and reduced EEG delta power were also observed in these pups, all of which indicated brain immaturity. All these signs of delayed maturation, usually found in premature babies, were observed in the pups of sleep restricted mothers, who had longer gestation period. This report not only shows the importance of sleep during pregnancy, but it also suggests that neonatal sleep monitoring can be used as a tool for early assessment of retarded brain development.

Sleep disorders in the elderly: a growing challenge.

In contrast to newborns, who spend 16-20 h in sleep each day, adults need only about sleep daily. However, many elderly may struggle to obtain those 8 h in one block. In addition to changes in sleep duration, sleep patterns change as age progresses. Like the physical changes that occur during old age, an alteration in sleep pattern is also a part of the normal ageing process. As people age, they tend to have a harder time falling asleep and more trouble staying asleep. Older people spend more time in the lighter stages of sleep than in deep sleep. As the circadian mechanism in older people becomes less efficient, their sleep schedule is shifted forward. Even when they manage to obtain 7 or 8 h sleep, they wake up early, as they have gone to sleep quite early. The prevalence of sleep disorders is higher among older adults. Loud snoring, which is more common in the elderly, can be a symptom of obstructive sleep apnoea, which puts a person at risk for cardiovascular diseases, headaches, memory loss, and depression. Restless legs syndrome and periodic limb movement disorder that disrupt sleep are more prevalent in older persons. Other common medical problems of old age such as hypertension diabetes mellitus, renal failure, respiratory diseases such as asthma, immune disorders, gastroesophageal reflux disease, physical disability, dementia, pain, depression, and anxiety are all associated with sleep disturbances.

Categories of Wistar Rats Based on Anxiety Traits: A Study Using Factor and Cluster Method.

BACKGROUND: Outcomes of neurobehavioral studies are invariably affected by the variations in the anxiety traits of animals in the same species. Identifying those traits and categorizing them accordingly would improve the reproducibility of results, and reduce the variation in the results from different laboratories. PURPOSE: The present study was done to identify the possible groups among the normal population of outbred adult male Wistar rats. METHODS: Anxiety traits were measured in elevated plus maze (EPM) test and open field test (OFT). The various anxiety responses from these tests were subjected to exploratory factor and hierarchical cluster analyses. Different clusters thus derived were compared with each other. RESULTS AND CONCLUSION: In exploratory factorial analysis, 2 components, that is, anxiety and activity were derived from the EPM and OFT parameters. Cluster analysis of EPM parameters classified the rats into 3 groups "high anxiety and low activity", "medium anxiety and high activity", and "low anxiety and medium activity". Whereas, cluster analysis on OFT parameters identified one more group namely "low anxiety and high activity". The rats which came under the clusters formed from the EPM and OFT parameters were not identical. Moreover, EPM and OFT may be measuring different aspects of anxiety.

Assessment of the septal area neuronal activity during penile erections in rapid eye movement sleep and waking in the rats.

To understand the central mechanism of penile erections during rapid eye movement (REM) sleep and waking, single units were recorded from the septal area in un-anesthetized head-restrained rats simultaneous with erections. Erectile events were assessed by pressure in the bulb of the corpus spongiosum of the penis and bulbospongiosus-muscle activity. Of 143 recorded neurons, 36% showed increased activity (E-type) and 24% decreased activity (I-type) during different phases of erection in REM sleep, while 10% were E-type and 35% were I-type during erections in waking. Most E-type neurons were recorded from the dorsal and intermediate part of lateral septum, whereas I-type neurons were from the medial septum. The findings illustrate the extensive network of various types of neurons in the septal area that fire in concert in relation to erection during REM sleep and waking. This study provides a unique prospective of the septal area for perpetuation of erectile circuitry during sleep.

The effects of rapid eye movement sleep deprivation during late pregnancy on newborns' sleep.

Sleep deprivation during pregnancy is an emerging concern, as it can adversely affect the development of the offspring brain. This study was conducted to evaluate the effects of deprivation of rapid eye movement (REM) sleep during the third term of pregnancy on the sleep-wake profiles of neonates in the Wistar rat model. Sleep-wake patterns were assessed through electrophysiological measures and behavioural observations during postnatal days 1-21 on pups born to REM sleep-deprived dams and control rats. Pups of REM sleep-deprived dams had active sleep that was not only markedly higher in percentage during all the days studied, but also had reduced latency during later postnatal days 15-21. Quiet sleep and wake periods were lower. These factors, along with less frequent but longer sleep-wake cycles, indicated maturational delay in the sleep-wake neural networks. The disruption of time-bound growth of sleep-wake neural networks was substantiated further by the decreased slope of survival plots in the sleep bouts. Examination of altered sleep-wake patterns during early development may provide crucial information concerning deranged neural development in the offspring. This is the first report, to our knowledge, to show that maternal sleep deprivation during pregnancy can delay and impair the development of sleep-wake profile in the offspring.

Sleep promoting potential of low dose α-Asarone in rat model.

Commonly used hypnotics have undesirable side-effects, especially during continuous usage. On the other hand, some herbal products, which are used for prolonged periods, are suggested to have a sleep inducing property, though the claims have not been validated scientifically. The hypnotic potential of α-Asarone, an active principle of Acorus species, was tested in the present study by first identifying the optimal dose of α-Asarone for improving sleep, followed by studies that evaluated the effect of repeated administration of this optimal dose for five days on sleep deprived rats. Of all the doses tested (2, 10, 40, 80 and 120 mg/kg), 10 mg/kg α-Asarone improved the quality of sleep, as indicated by an increased NREM bout duration, reduced arousal index, and decreased bout frequencies of NREM sleep and wakefulness. A marginal decrease in the hypothalamic and body temperatures was also observed. Higher doses, on the other hand, not only reduced the quantity and quality of sleep, but also produced hypothermia. In sleep deprived rats, administration of 10 mg/kg α-Asarone for five consecutive days improved the quality of sleep in contrast to the vehicle and a known hypnotic midazolam. Improvement in NREM sleep quality was observed when the difference between the hypothalamic and the body temperature was minimum. An enhanced association between NREM sleep bout duration and hypothalamic temperature was also observed after administration of 10 mg/kg α-Asarone. This comprehensive study is the first report on the hypnotic property of α-Asarone, which validates its potential to be considered for treatment of insomnia.

Dynamic changes in sleep pattern during post-partum in normal pregnancy in rat model.

To develop an animal model for studies on peri-partum sleep disorders, sleep patterns in female Wistar rats during pregnancy, post-partum and after weaning, were assessed and associated adaptive changes in their anxiety were examined. Adult nulliparous female rats, maintained in standard laboratory conditions with ad libitum food and water, were surgically implanted with electroencephalogram and electromyogram electrodes under anaesthesia for objective assessment of sleep-wakefulness (S-W). After post-surgical recovery, three control recordings of S-W were taken for 24h before the animals were kept for mating. After confirmation of pregnancy, S-W recordings were acquired during different days of pregnancy, post-partum lactation/nursing days, and also after weaning. Their anxiety levels were tested in the elevated plus maze. During pregnancy, sleep increased primarily due to increase in light non-REM sleep during dark period. There was an increase in non-REM sleep delta power after parturition, though the sleep was fragmented, especially during daytime. Simultaneous behavioural recording showed increased anxiety during third trimester of pregnancy and gradual reversal of it after parturition. This is the first report where diurnal and nocturnal variations in S-W and delta power, along with adaptive changes in anxiety, were studied before, during and after pregnancy. This study also provides an animal model for drug trials and studies on sleep disorders during peri-partum window.

Increased ultrasonic vocalizations and risk-taking in rat pups of sleep-deprived dams.

Ultrasonic vocalizations (USVs) in rodent pups are analogous to cries in human babies. There is reduction in USVs in pups after experimental deprivation of rapid eye movement sleep of dams during pregnancy. However, the effects of total sleep deprivation on the USVs of newborns and their emotional development are not documented. Male pups born to the rats that underwent total sleep deprivation for 5h during the third trimester made higher vocalizations, when tested on early postnatal days (pnds) in an isolation-paradigm. Their anxiety-related behaviors during pnds 25-28, were tested using elevated plus maze (EPM). In comparison to the control pups, weanlings of sleep-deprived dams made increased entries into the open arms and higher mobility in the EPM. Enhanced distress calls during early pnds and reduction in risk assessment in weanlings indicate a link between the two behaviors. The USVs during ontogeny may provide early signals about altered emotional development.

Sleep deprivation during late pregnancy produces hyperactivity and increased risk-taking behavior in offspring.

Sleep deprivation in women resulting from their modern lifestyle, especially during pregnancy, is a serious concern as it can affect the health of the newborn. Anxiety disorders and cognitive deficits in the offspring are also on the rise. However, experimental studies on the effects of sleep loss during pregnancy, on emotional development and cognitive function of the newborn, are scanty in literature. In the current study, female rats were sleep-deprived for 5h by gentle handling, during the 6 days of the third trimester (days 14-19 of pregnancy). The effects of this sleep deprivation on anxiety-related behaviors of pups during their peri-adolescence age were studied using elevated plus maze (EPM). In addition to body weights of dams and offspring, the maternal behavior was also monitored. The weanlings of sleep-deprived dams showed heightened risk-taking behavior as they made increased explorations into the open arms of EPM. They also showed higher mobility in comparison to the control group. Though the body weights of sleep-deprived dams were comparable to those of the control group, their newborns had lower birth weight. Nevertheless, these pups gained weight and reached the control group values during the initial post-natal week. But after weaning, their rate of growth was lower than that of the control group. This is the first report providing evidences for the role of sleep during late pregnancy in shaping the neuropsychological development in offspring.

Reduction in ultrasonic vocalizations in pups born to rapid eye movement sleep restricted mothers in rat model.

The effects of rapid eye movement sleep restriction (REMSR) in rats during late pregnancy were studied on the ultrasonic vocalizations (USVs) made by the pups. USVs are distress calls inaudible to human ears. Rapid eye movement (REM) sleep was restricted in one group of pregnant rats for 22 hours, starting from gestational day 14 to 20, using standard single platform method. The USVs of male pups were recorded after a brief isolation from their mother for two minutes on alternate post-natal days, from day one till weaning. The USVs were recorded using microphones and were analysed qualitatively and quantitatively using SASPro software. Control pups produced maximum vocalization on post-natal days 9 to 11. In comparison, the pups born to REMSR mothers showed not only a reduction in vocalization but also a delay in peak call making days. The experimental group showed variations in the types and characteristics of call types, and alteration in temporal profile. The blunting of distress call making response in these pups indicates that maternal sleep plays a role in regulating the neural development involved in vocalizations and possibly in shaping the emotional behaviour in neonates. It is suggested that the reduced ultrasonic vocalizations can be utilized as a reliable early marker for affective state in rat pups. Such impaired vocalization responses could provide an important lead in understanding mother-child bonding for an optimal cognitive development during post-partum life. This is the first report showing a potential link between maternal REM sleep deprivation and the vocalization in neonates and infants.