Biological sex influences sleep phenotype in mice experiencing spontaneous opioid withdrawal.

Aversive symptoms, including insomnia experienced during opioid withdrawal, are a major drive to relapse; however, withdrawal-associated sleep symptomatology has been little explored in preclinical models. We describe here a model of opioid withdrawal in mice that resembles the sleep phenotype characteristic of withdrawal in humans. Male and female C57BL/6 mice were instrumented with telemeters to record electroencephalogram, electromyogram, activity and subcutaneous temperature. All mice received two treatments separated by a 16-day washout period: (1) saline (volume: 10 ml kg-1 ); or (2) ascending doses of morphine (5, 10, 20, 40 and 80 mg kg-1 ; volume: 10 ml kg-1 ) for 5 days at Zeitgeber time 1 and Zeitgeber time 13. Recordings for the first 71 hr after treatment discontinuation (withdrawal days 1-3) and for 24 hr on withdrawal days 5 and 7 were scored for sleep/wake state, and sleep architecture and electroencephalogram spectral data were analysed. Morphine was acutely wake- and activity-promoting, and non-rapid eye movement and rapid eye movement sleep were increased during the dark phase on withdrawal day 2 in both sexes. While non-rapid eye movement delta power (0.5-4.0 Hz), a measure of sleep intensity, was reduced during the light phase on withdrawal day 1 and the dark phase on withdrawal day 2 in both sexes, female mice also exhibited changes in the duration and the number of bouts of sleep/wake states. These observations of fragmented sleep on withdrawal days 1-3 suggest poorer sleep consolidation and a more pronounced withdrawal-associated sleep phenotype in female than in male mice. These data may indicate a greater sensitivity to morphine, a more distinct aversive sleep phenotype and/or a faster escalation to dependence in female mice.

Evaluation of the efficacy of the hypocretin/orexin receptor agonists TAK-925 and ARN-776 in narcoleptic orexin/tTA; TetO-DTA mice.

The sleep disorder narcolepsy, a hypocretin deficiency disorder thought to be due to degeneration of hypothalamic hypocretin/orexin neurons, is currently treated symptomatically. We evaluated the efficacy of two small molecule hypocretin/orexin receptor-2 (HCRTR2) agonists in narcoleptic male orexin/tTA; TetO-DTA mice. TAK-925 (1-10 mg/kg, s.c.) and ARN-776 (1-10 mg/kg, i.p.) were injected 15 min before dark onset in a repeated measures design. EEG, EMG, subcutaneous temperature (Tsc ) and activity were recorded by telemetry; recordings for the first 6 h of the dark period were scored for sleep/wake and cataplexy. At all doses tested, TAK-925 and ARN-776 caused continuous wakefulness and eliminated sleep for the first hour. Both TAK-925 and ARN-776 caused dose-related delays in NREM sleep onset. All doses of TAK-925 and all but the lowest dose of ARN-776 eliminated cataplexy during the first hour after treatment; the anti-cataplectic effect of TAK-925 persisted into the second hour for the highest dose. TAK-925 and ARN-776 also reduced the cumulative amount of cataplexy during the 6 h post-dosing period. The acute increase in wakefulness produced by both HCRTR2 agonists was characterised by increased spectral power in the gamma EEG band. Although neither compound provoked a NREM sleep rebound, both compounds affected NREM EEG during the second hour post-dosing. TAK-925 and ARN-776 also increased gross motor activity, running wheel activity, and Tsc , suggesting that the wake-promoting and sleep-suppressing activities of these compounds could be a consequence of hyperactivity. Nonetheless, the anti-cataplectic activity of TAK-925 and ARN-776 is encouraging for the development of HCRTR2 agonists.

Wakefulness Induced by TAAR1 Partial Agonism is Mediated Through Dopaminergic Neurotransmission.

Trace amine-associated receptor 1 (TAAR1) is known to negatively regulate dopamine (DA) release. The partial TAAR1 agonist RO5263397 promotes wakefulness and suppresses NREM and REM sleep in mice, rats, and non-human primates. We tested the hypothesis that the TAAR1-mediated effects on sleep/wake were due, at least in part, to DA release. Male C57BL6/J mice (n=8) were intraperitoneally administered the D1R antagonist SCH23390, the D2R antagonist eticlopride, a combination of D1R+D2R antagonists or saline at ZT5.5, followed 30 min later by RO5263397 or vehicle (10% DMSO in DI water) at ZT6 per os. EEG, EMG, subcutaneous temperature, and activity were recorded in each mouse across the 8 treatment conditions and sleep architecture was analyzed for 6 hours post-dosing. Consistent with our previous reports, RO5263397 increased wakefulness as well as the latency to NREM and REM sleep. D1, D2, and D1+D2 pretreatment reduced RO5263397-induced wakefulness during the first 1-2 hours after dosing, but only the D1+D2 combination attenuated the wake-promoting effect of RO5263397 from ZT6-8, mostly by increasing NREM sleep. Although D1+D2 antagonism blocked the wake-promoting effect of RO5263397, only the D1 antagonist significantly reduced the TAAR1-mediated increase in NREM latency. Neither the D1 nor the D2 antagonist affected TAAR1-mediated suppression of REM sleep. These results suggest that, whereas TAAR1 effects on wakefulness are mediated in part through the D2R, D1R activation plays a role in reversing the TAAR1-mediated increase in NREM sleep latency. By contrast, TAAR1-mediated suppression of REM sleep appears not to involve D1R or D2R mechanisms.

An implantable neural sensing microsystem with fiber-optic data transmission and power delivery.

We have developed a prototype cortical neural sensing microsystem for brain implantable neuroengineering applications. Its key feature is that both the transmission of broadband, multichannel neural data and power required for the embedded microelectronics are provided by optical fiber access. The fiber-optic system is aimed at enabling neural recording from rodents and primates by converting cortical signals to a digital stream of infrared light pulses. In the full microsystem whose performance is summarized in this paper, an analog-to-digital converter and a low power digital controller IC have been integrated with a low threshold, semiconductor laser to extract the digitized neural signals optically from the implantable unit. The microsystem also acquires electrical power and synchronization clocks via optical fibers from an external laser by using a highly efficient photovoltaic cell on board. The implantable unit employs a flexible polymer substrate to integrate analog and digital microelectronics and on-chip optoelectronic components, while adapting to the anatomical and physiological constraints of the environment. A low power analog CMOS chip, which includes preamplifier and multiplexing circuitry, is directly flip-chip bonded to the microelectrode array to form the cortical neurosensor device.

The development of sleep/wake disruption and cataplexy as hypocretin/orexin neurons degenerate in male vs. female Orexin/tTA; TetO-DTA Mice.

Narcolepsy Type 1 (NT1), a sleep disorder with similar prevalence in both sexes, is thought to be due to loss of the hypocretin/orexin (Hcrt) neurons. Several transgenic strains have been created to model this disorder and are increasingly being used for preclinical drug development and basic science studies, yet most studies have solely used male mice. We compared the development of narcoleptic symptomatology in male vs. female orexin-tTA; TetO-DTA mice, a model in which Hcrt neuron degeneration can be initiated by removal of doxycycline (DOX) from the diet. EEG, EMG, subcutaneous temperature, gross motor activity, and video recordings were conducted for 24-h at baseline and 1, 2, 4, and 6 weeks after DOX removal. Female DTA mice exhibited cataplexy, the pathognomonic symptom of NT1, by Week 1 in the DOX(-) condition but cataplexy was not consistently present in males until Week 2. By Week 2, both sexes showed an impaired ability to sustain long wake bouts during the active period, the murine equivalent of excessive daytime sleepiness in NT1. Subcutaneous temperature appeared to be regulated at lower levels in both sexes as the Hcrt neurons degenerated. During degeneration, both sexes also exhibited the "Delta State", characterized by sudden cessation of activity, high delta activity in the EEG, maintenance of muscle tone and posture, and the absence of phasic EMG activity. Since the phenotypes of the two sexes were indistinguishable by Week 6, we conclude that both sexes can be safely combined in future studies to reduce cost and animal use.

Listening to Brain Microcircuits for Interfacing With External World-Progress in Wireless Implantable Microelectronic Neuroengineering Devices: Experimental systems are described for electrical recording in the brain using multiple microelectrodes and short range implantable or wearable broadcasting units.

Acquiring neural signals at high spatial and temporal resolution directly from brain microcircuits and decoding their activity to interpret commands and/or prior planning activity, such as motion of an arm or a leg, is a prime goal of modern neurotechnology. Its practical aims include assistive devices for subjects whose normal neural information pathways are not functioning due to physical damage or disease. On the fundamental side, researchers are striving to decipher the code of multiple neural microcircuits which collectively make up nature's amazing computing machine, the brain. By implanting biocompatible neural sensor probes directly into the brain, in the form of microelectrode arrays, it is now possible to extract information from interacting populations of neural cells with spatial and temporal resolution at the single cell level. With parallel advances in application of statistical and mathematical techniques tools for deciphering the neural code, extracted populations or correlated neurons, significant understanding has been achieved of those brain commands that control, e.g., the motion of an arm in a primate (monkey or a human subject). These developments are accelerating the work on neural prosthetics where brain derived signals may be employed to bypass, e.g., an injured spinal cord. One key element in achieving the goals for practical and versatile neural prostheses is the development of fully implantable wireless microelectronic "brain-interfaces" within the body, a point of special emphasis of this paper.

Focused Ultrasound for Noninvasive, Focal Pharmacologic Neurointervention.

A long-standing goal of translational neuroscience is the ability to noninvasively deliver therapeutic agents to specific brain regions with high spatiotemporal resolution. Focused ultrasound (FUS) is an emerging technology that can noninvasively deliver energy up the order of 1 kW/cm2 with millimeter and millisecond resolution to any point in the human brain with Food and Drug Administration-approved hardware. Although FUS is clinically utilized primarily for focal ablation in conditions such as essential tremor, recent breakthroughs have enabled the use of FUS for drug delivery at lower intensities (i.e., tens of watts per square centimeter) without ablation of the tissue. In this review, we present strategies for image-guided FUS-mediated pharmacologic neurointerventions. First, we discuss blood-brain barrier opening to deliver therapeutic agents of a variety of sizes to the central nervous system. We then describe the use of ultrasound-sensitive nanoparticles to noninvasively deliver small molecules to millimeter-sized structures including superficial cortical regions and deep gray matter regions within the brain without the need for blood-brain barrier opening. We also consider the safety and potential complications of these techniques, with attention to temporal acuity. Finally, we close with a discussion of different methods for mapping the ultrasound field within the brain and describe future avenues of research in ultrasound-targeted drug therapies.

Novel natural fill telemetric pressure flow study of discomfort and bladder outlet obstruction.

PURPOSE: We evaluated the accuracy of natural fill telemetric pressure flow study performed in a private room, and assessed patient discomfort and experience after the procedure compared to those of standard pressure flow study. MATERIALS AND METHODS: In 58 patients with lower urinary tract symptoms/benign prostatic hyperplasia free uroflowmetry, and natural fill telemetric and standard pressure flow studies were prospectively performed. Immediately after each step patients were asked to rate the experience in terms of pain, embarrassment, bother, boredom and repeat testing. Subjective items and objective urodynamic parameters were compared among the 3 tests. RESULTS: Maximum urine flow on natural fill telemetric pressure flow study was not different from that on free uroflowmetry. In contrast, maximum flow, detrusor pressure at maximum flow and bladder contractility index on the standard pressure flow study were significantly lower than on the natural fill pressure flow study. However, the bladder outlet obstruction index was not significantly different between the 2 studies, and the bladder outlet obstruction and bladder contractility indexes correlated well. There were also no differences in bother and embarrassment scores. However, natural fill telemetric pressure flow study was superior in terms of pain and boredom scores. Patients were more willing to undergo repeat natural fill telemetric pressure flow study than standard pressure flow study. Time to complete the test was significantly shorter for the natural fill study than for the standard study. CONCLUSIONS: Flow rate differences between pressure flow studies and free uroflowmetry are not due to mechanical obstruction by the catheter but to other factors, such as the bladder filling method. Objective parameters on the natural fill telemetric pressure flow study correlated with standard pressure flow study findings. The natural fill telemetric pressure flow study shortened the time needed for the test and may decrease the discomfort of the standard pressure flow study.

Effect of superheated steam roasting with hot smoking treatment on improving physicochemical properties of the adductor muscle of pen shell (Atrina pectinate).

The adductor muscle of the pen shell Atrina pectinata (AMPS) is a popular protein-enriched food item in Asian Pacific countries, and has only been marketed in the frozen condition, as a result browning and decreased sensory attributes occur. To overcome these problems, superheated steam roasting (at 270°C for 4 min) combined with the hot smoke (10 min) using a selective Oak sawdust was employed to develop a new AMPS product yielding high physicochemical properties during storage periods (0-13 days) especially at 10°C. The processed AMPS showed high sensory preferences because of good odor, color, and textural properties. It also significantly inhibited bacterial growth, volatile basic nitrogen, thiobarbituric acid-reactive species, and pH changes, and eventually possessed higher nutritional composition with low trimethylamine N-oxide level. Results indicate that saturated steam allows AMPS at good physicochemical conditions, whereas hot smoke-derived aroma compounds prolong its shelf life through antioxidant and antimicrobial activities.

The antioxidant properties of brown seaweed (Sargassum siliquastrum) extracts.

Antioxidant properties of brown seaweed (Sargassum siliquastrum) extracts were evaluated using various antioxidant measurements, i.e., inhibitory effect on thiobarbituric acid-reactive substances (TBARS), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, metal chelating effect, reducing power effect, and total phenolic compounds. When the extraction solvents n-hexane, chloroform, ethanol, and water were compared, the water extract showed the highest yield in extracted mass. Total phenolic compounds were the highest in the ethanol extract, with 127.4 mg/g. The TBARS inhibition of chloroform and ethanol extracts at 10 mg/mL was 90.9% and 80.9%, respectively. DPPH radical scavenging capacity was more than 90% in all extracts at 1 mg/mL. The chloroform extract exhibited the highest metal ion chelating ability of 69.6% at 10 mg/mL. The reducing power was found to be the highest in the ethanol extract at 10 mg/mL, showing an effect similar to ascorbic acid. Thus, the ethanol extract of S. siliquastrum has potential as a natural antioxidant.

Peripheral optogenetic stimulation induces whisker movement and sensory perception in head-fixed mice.

We discovered that optical stimulation of the mystacial pad in Emx1-Cre;Ai27D transgenic mice induces whisker movements due to activation of ChR2 expressed in muscles controlling retraction and protraction. Using high-speed videography in anesthetized mice, we characterize the amplitude of whisker protractions evoked by varying the intensity, duration, and frequency of optogenetic stimulation. Recordings from primary somatosensory cortex (S1) in anesthetized mice indicated that optogenetic whisker pad stimulation evokes robust yet longer latency responses than mechanical whisker stimulation. In head-fixed mice trained to report optogenetic whisker pad stimulation, psychometric curves showed similar dependence on stimulus duration as evoked whisker movements and S1 activity. Furthermore, optogenetic stimulation of S1 in expert mice was sufficient to substitute for peripheral stimulation. We conclude that whisker protractions evoked by optogenetic activation of whisker pad muscles results in cortical activity and sensory perception, consistent with the coding of evoked whisker movements by reafferent sensory input.

Pathogenic microorganisms, heavy metals, and antibiotic residues in seven Korean freshwater aquaculture species.

This survey was performed to estimate the levels of pathogenic microorganisms, antibiotic residues, and heavy metals in seven Korean freshwater aquaculture species including Anguilla japonica, Cyprinus carpio nudus, Oncorhynchus mykiss, Pseudobagrus fulvidraco, Semisulcospira coreana, Silurus asotus, and Trionyxs sinensis. None of the ten foodborne pathogens tested in this study were found in any of the species collected from any of the aquaculture farms. Furthermore, no banned chemicals or antibiotic residues were found in any of the species collected from any of the aquaculture farms, except enrofloxacin, which was below guideline limits (0.1 mg/kg). Finally, no species had lead, cadmium, total arsenic, or total mercury concentrations above the Ministry of Food and Drug Safety (MSDF) guidelines (0.5, 0.5, 0.1, and 0.5 mg/kg, respectively). These results ensure the safety of freshwater aquaculture species and will be useful for developing consumption advisories of freshwater fishes.

Oral Administration of a Hot Water Extract of the Softshell Turtle (Trionyx sinensis) Improves Exercise Performance.

Freshwater softshell turtle (Trionyx sinensis) extract has been used traditionally as a tonic soup, and to recover from physical fatigue. To support these claims, the forelimb grip strength of mice was measured after feeding a soft-shell turtle extract for 7 days. The T. sinensis extract significantly increased the grip strength to 1.25±0.07 N (P<0.01), which is 16.8% higher than the force on day 0. After exercising, the blood glucose levels in extract-fed mice were 202% higher and urea levels were 73% lower, which were both significantly different than the levels observed after control treatment. Lactate dehydrogenase was significantly higher by 314%, and glutathione peroxidase increased by 165%. In addition, the obesity markers, serum triglyceride and cholesterol, decreased to 62% and 49%, respectively, after mice were fed the extract. These data show that the T. sinensis extract provided more energy for forelimb exercise, prevented protein catabolism and muscle fatigue, and decreased the oxidative stress caused by an exhaustive workout.

Hot Water Extract of Leather Carp (Cyprinus carpio nudus) Improves Exercise Performance in Mice.

The hot water extract of leather carp (Cyprinus carpio nudus) has been used as a nourishing tonic soup and as an aid for recovery from physical fatigue. In this study, we investigated the effect of leather carp extract on exercise performance in mice. Swimming endurance and forelimb grip strength were assessed following oral administration of the extract (once per day for 7 days) at a dose of 0.5 mg/10 µL/g body weight. After 7 days, mice given the leather carp extract had significantly greater swimming endurance [105±18 s (P<0.05); 52% longer than day 0] and forelimb grip strength [1.18±0.05 Newton (P<0.01); 17% greater than day 0]. The extract increased muscle mass, but had little effect on body weight. Following the swimming exercise, blood glucose, glutathione peroxidase, and superoxide dismutase levels in extract-fed mice were significantly higher (145%, 131%, and 106%, respectively) than in the saline control group. Blood levels of high-density lipoprotein cholesterol were also significantly increased (128%) in mice given the extract compared to the controls. These results suggest that leather carp extract can improve physical exercise performance and prevent oxidative stress caused by exhaustive workouts.

Characteristics and in vitro Anti-diabetic Properties of the Korean Rice Wine, Makgeolli Fermented with Laminaria japonica.

New in vitro anti-diabetes makgeolli was produced from rice by adding various quantities of Laminaria japonica, and the fermentation characteristics of the L. japonica makgeolli during the fermentation process were investigated. The contents of alcohol and reducing sugar, and viable count of yeast, of L. japonica makgeolli were not significantly changed when the proportion of L. japonica was increased. The total acid content decreased with an increase in L. japonica concentration; the pH and total bacterial cell count increased in proportion with the increase in L. japonica concentration. The L. japonica makgeolli contents of free sugars, such as fructose, glucose, and sucrose, and of organic acids, such as acetic acid, citric acid, succinic acid, and lactic acid, were altered during fermentation and showed various patterns. The effects of the quantity of L. japonica added on the acceptability and anti-diabetes activities of L. japonica makgeolli were also investigated. In a sensory evaluation, L. japonica makgeolli brewed by adding 2.5 or 5% L. japonica to the mash showed the best overall acceptability; the 12.5% L. japonica sample was least favored due to its seaweed flavor. L. japonica addition did not increase the peroxynitrite-scavenging activity of makgeolli. L. japonica makgeolli showed potent anti-diabetes activity, particularly that containing >7.5% L. japonica. Therefore, L. japonica makgeolli may represent a new functional makgeolli with anti-diabetes properties.

Antibacterial effect of grapefruit seed extract (GSE) on Makgeolli-brewing microorganisms and its application in the preservation of fresh Makgeolli.

UNLABELLED: To develop a new preservation method, the antimicrobial activity of grapefruit seed extract (GSE) against Makgeolli-brewing microorganisms and food-borne pathogens was assessed, and a general analysis and sensory evaluation of fresh Makgeolli with added GSE was made. The minimum inhibitory concentration (MIC) values of GSE against 10 strains of Makgeolli-brewing microorganism were 0.0122 to 1.5625 µL/mL. The MIC values against 6 strains of food-borne pathogens were 0.0061 to 0.7813 µL/mL. On addition of 0.1% (v/v) and 0.2% GSE in bottled fresh Makgeolli, no significant difference in the pH, or the contents of total acids, ethanol, or methanol in the Makgeolli, were observed compared with control Makgeolli (with no GSE), during the preservation period (8 weeks) at 10 °C. In the Makgeolli with 0.1% and 0.2% GSE, the total bacterial counts decreased significantly by 4.9% (P < 0.01) and 11.2% (P < 0.001), respectively, versus the control. The decreases in yeast count were significantly lessened by 15.33% and 15.24% (both P < 0.001), respectively, after 8 weeks of storage, compared with the control. In the sensory evaluation of Makgeolli with 0.1% and 0.2% GSE, the refreshment and overall acceptability received significantly better scores than the control (P < 0.01), with no change in sweetness, bitterness, sourness, turbidity, color, or odor. These results suggest that GSE controls the growth of Makgeolli-brewing microorganisms and extends the shelf life (ca. 2 wk), without decreasing overall acceptance. PRACTICAL APPLICATION: A new preservation method for fresh Makgeolli by adding grapefruit seed extract (GSE) was developed. As fresh Makgeolli contains live microorganisms, the preservation period is 1 wk, which is relatively short. GSE controls the growth of Makgeolli-brewing and Makgeolli-spoiling microorganisms. 0.1% to 0.2% GSE is optimum for prolonging the shelf life (2 wk) of bottled fresh Makgeolli, and has no adverse effect on overall acceptability. We demonstrated that GSE is an effective natural additive that prolongs the shelf life of fresh Makgeolli with no significant loss in quality.

Wireless, high-bandwidth recordings from non-human primate motor cortex using a scalable 16-Ch implantable microsystem.

A multitude of neuroengineering challenges exist today in creating practical, chronic multichannel neural recording systems for primate research and human clinical application. Specifically, a) the persistent wired connections limit patient mobility from the recording system, b) the transfer of high bandwidth signals to external (even distant) electronics normally forces premature data reduction, and c) the chronic susceptibility to infection due to the percutaneous nature of the implants all severely hinder the success of neural prosthetic systems. Here we detail one approach to overcome these limitations: an entirely implantable, wirelessly communicating, integrated neural recording microsystem, dubbed the Brain Implantable Chip (BIC).