Essential Roles of GABA Transporter-1 in Controlling Rapid Eye Movement Sleep and in Increased Slow Wave Activity after Sleep Deprivation

GABA is the major inhibitory neurotransmitter in the mammalian central nervous system that has been strongly implicated in the regulation of sleep. GABA transporter subtype 1 (GAT1) constructs high affinity reuptake sites for GABA and regulates GABAergic transmission in the brain. However, the role of GAT1 in sleep-wake regulation remains elusive. In the current study, we characterized the spontaneous sleep-wake cycle and responses to sleep deprivation in GAT1 knock-out (KO) mice. GAT1 KO mice exhibited dominant theta-activity and a remarkable reduction of EEG power in low frequencies across all vigilance stages. Under baseline conditions, spontaneous rapid eye movement (REM) sleep of KO mice was elevated both during the light and dark periods, and non-REM (NREM) sleep was reduced during the light period only. KO mice also showed more state transitions from NREM to REM sleep and from REM sleep to wakefulness, as well as more number of REM and NREM sleep bouts than WT mice. During the dark period, KO mice exhibited more REM sleep bouts only. Six hours of sleep deprivation induced rebound increases in NREM and REM sleep in both genotypes. However, slow wave activity, the intensity component of NREM sleep was briefly elevated in WT mice but remained completely unchanged in KO mice, compared with their respective baselines. These results indicate that GAT1 plays a critical role in the regulation of REM sleep and homeostasis of NREM sleep.     

Circulating miRNA-33: a potential biomarker in patients with coronary artery disease

Background: Circulating microRNAs (miRNA) are present in body fluids in stable, cell-free form. Likewise, these miRNAs can be identified in various stages of coronary artery disease (CAD) such as inflammation, endothelial dysfunction, proliferation and atherosclerosis among others. miRNA expression levels can be identified.

Aims and objectives: To determine the expression of circulating miRNAs (miR-126, miR-92, miR-33, miR-145 and miR-155) in CAD patients of Indian origin.

Material and methods: miRNA profiling analysis in blood plasma was performed by quantitative real-time-PCR (qRT-PCR) in 60 angiographically verified subjects including 30 CAD patients and 30 age- and gender-matched controls. Association between the expression of all five circulating miRNAs and clinical characteristics of patients with CAD were analysed using Medcalc statistics. The severity of CAD was assessed using SYNTAX score (SS).

Results: Expression of plasma miR-33 increased by 2.9 folds in CAD patients than in control group (p value ≥0.002) also it was found that miR-33 expression levels in mild cases (SS: ≤22) were significantly higher than CAD controls. There was a modest negative correlation between miR-33 and total cholesterol/high density lipoprotein ratio, triglycerides and very low density lipoprotein.

Conclusion: The study reports a significant association between increased levels of plasma miR-33 and CAD. Thus, plasma miR-33 appears to be a promising non-invasive biomarker, but requires further validation in a large cohort.     

Is C-reactive protein elevated in obstructive sleep apnea? a systematic review and meta-analysis

Purpose: This study examined whether circulating C-reactive protein (CRP) is elevated in obstructive sleep apnoea (OSA) independent of the confounding effects of comorbidities, smoking, body mass index (BMI), age and gender.

Methods: A systematic review of the literature was performed using PubMed, Embase and Cochrane databases from 1 January 1997 to 1 November 2017 using the key words obstructive sleep apnoea and C-Reactive protein to identify full text English language studies that compared CRP in adult non-smoking OSA participants without comorbidities and adult healthy non-smoking control participants matched for BMI, age and gender. Data from eligible studies were subjected to meta-analysis using RevMan version 5.3.

Results: Five studies (219 OSA participants, 116 controls) met the selection criteria. The total standard mean difference for circulating high sensitivity CRP was 0.61?mg/dL higher in OSA participants than in control participants (confidence interval: 0.38 to 0.84, p?<?0.00001), with low between-studies heterogeneity (df = 7, p?=?0.16, I²?=?33%) and minimal evidence of publication bias.

Conclusions: CRP levels in non-smoking OSA participants without comorbidities were increased relative to levels in healthy matched non-smoking control participants, suggesting that pharyngeal or systemic inflammatory effects attributable to OSA may elevate CRP.     

Altered sleep regulation in leptin-deficient mice

Recent epidemiological, clinical, and experimental studies have demonstrated important links between sleep duration and architecture, circadian rhythms, and metabolism, although the genetic pathways that interconnect these processes are not well understood. Leptin is a circulating hormone and major adiposity signal involved in long-term energy homeostasis. In this study, we tested the hypothesis that leptin deficiency leads to impairments in sleep-wake regulation. Male ob/ob mice, a genetic model of leptin deficiency, had significantly disrupted sleep architecture with an elevated number of arousals from sleep [wild-type (WT) mice, 108.2 +/- 7.2 vs. ob/ob mice, 148.4 +/- 4.5, P < 0.001] and increased stage shifts (WT, 519.1 +/- 25.2 vs. ob/ob, 748.0 +/- 38.8, P < 0.001) compared with WT mice. Ob/ob mice also had more frequent, but shorter-lasting sleep bouts compared with WT mice, indicating impaired sleep consolidation. Interestingly, ob/ob mice showed changes in sleep time, with increased amounts of 24-h non-rapid eye movement (NREM) sleep (WT, 601.5 +/- 10.8 vs. ob/ob, 669.2 +/- 13.4 min, P < 0.001). Ob/ob mice had overall lower body temperature (WT, 35.1 +/- 0.2 vs. ob/ob, 33.4 +/- 0.2 degrees C, P < 0.001) and locomotor activity counts (WT, 25125 +/- 2137 vs. ob/ob, 5219 +/- 1759, P < 0.001). Ob/ob mice displayed an attenuated diurnal rhythm of sleep-wake stages, NREM delta power, and locomotor activity. Following sleep deprivation, ob/ob mice had smaller amounts of NREM and REM recovery sleep, both in terms of the magnitude and the duration of the recovery response. In combination, these results indicate that leptin deficiency disrupts the regulation of sleep architecture and diurnal rhythmicity.     

Circulating miRs-183-5p, -206-3p and -381-3p may serve as novel biomarkers for 4,4’-methylene diphenyl diisocyanate exposure

Background: Occupational exposure to the most widely used diisocyanate, 4,4’-methylene diphenyl diisocyanate (MDI), is a cause of occupational asthma (OA). Early recognition of MDI exposure and sensitization is essential for the prevention of MDI-OA.

Objective: Identify circulating microRNAs (miRs) as novel biomarkers for early detection of MDI exposure and prevention of MDI-OA.

Materials and methods: Female BALB/c mice were exposed to one of three exposure regimens: dermal exposure to 1% MDI in acetone; nose-only exposure to 4580?±?1497?μg/m³ MDI-aerosol for 60?minutes; or MDI dermal exposure/sensitization followed by MDI-aerosol inhalation challenge. Blood was collected and miRCURY? miRs qPCR Profiling Service was used to profile circulate miRs from dermally exposed mice. Candidate miRs were identified and verified from mice exposed to three MDI-exposure regimens by TaqMan^® miR assays.

Results: Up/down-regulation patterns of circulating mmu-miRs-183-5p, -206-3p and -381-3p were identified and verified. Circulating mmu-miR-183-5p was upregulated whereas mmu-miRs-206-3p and -381-3p were downregulated in mice exposed via all three MDI exposure regimens.

Discussion and conclusion: Upregulation of circulating miR-183-5p along with downregulation of circulating miRs-206-3p and -381-3p may serve as putative biomarkers of MDI exposure and may be considered as potential candidates for validation in exposed human worker populations.     

Seasonal effects on the sleep–wake cycle,the rest–activity rhythm and quality of life for Japanese and Thai older people

Background: The sleep-wake cycle and the rest–activity rhythm are known to change with aging, and such changes have been implicated in higher levels of depression as well as an increased incidence of dementia. However, information supporting seasonal changes in the sleep–wake cycle, the rest–activity rhythm and quality of life in older community-dwelling people remains insufficient. The aim of the present study was to prospectively investigate seasonal effects on the sleep–wake cycle, the rest–activity rhythm and quality of life among older people living in areas of Japan or Thailand with different climate classifications.

Method: The survey was conducted from March 2016 to May 2017, and 109 participants were recruited from Japan and Thailand: 47 older people living in Akita prefecture, Japan, and 62 older people living in Chiang Mai or Nakhon Ratchasima, Thailand. According to the Köppen–Geiger classification of Asian climates comprising tropical, desert, steppe, temperate and subarctic climates, Akita prefecture, which is located in northern Japan, is classified as a humid subarctic climate, while the Thai study areas are classified as tropical savanna. To monitor parameters of the sleep–wake cycle during nighttime (e.g. total sleep time, sleep latency, sleep efficiency, awaking time and frequency of sleep interruptions) and to calculate parameters of the rest–activity rhythm over the 24 h profile (e.g., interdaily stability, intradaily variability, relative amplitude, mean of least active 5 h period and mean of most active 10 h period), all the participants from both countries wore an Actiwatch 2 device on their nondominant wrist continuously for 7 days during each local season. The World Health Organization Quality of Life Questionnaire-BREF (WHOQOL-BREF) was also assessed during each local season.

Results: The final sample size was 37 older people living in Akita prefecture, Japan, and 44 older people living in Thailand; these subjects completed the data collections during each local season. The dropout rates were 21% in Japan and 29% in Thailand. The results for the Japanese subjects showed a significantly shorter sleep time with higher levels of activity during the nighttime on summer (p < 0.001) and a fragmented rest–activity rhythm over the 24 h profile on winter (p < 0.001). The older Thai participants exhibited a poor state of night sleeping year-round, and a significant relationship was observed between seasonal variations in motor activity and the social domain of WHOQOL-BREF for each Thai season (|r| = 0.4, p < 0.01).

Conclusion: These findings provide new and important information regarding seasonal effects on the sleep–wake cycle, the rest–activity rhythm and quality of life in older community-dwelling people living in two different Asian climates. Consequently, clinical preventions targeting such seasonal variations might be useful for improving the quality of life of older Japanese and Thai individuals.     

miR-200b-3p in plasma is a potential diagnostic biomarker in oral squamous cell carcinoma

Context: Circulating MicroRNAs (miRNAs) are emerging as novel biomarkers for tumour.

Objective: Evaluate the diagnostic potential of plasma miR-200b-3p in oral squamous cell carcinoma (OSCC).

Materials and methods: miR-200b-3p was detected by qRT-PCR in paired pre-operative and post-operative plasmas from 80 OSCC patients and 80 healthy controls.

Results: Plasma miR-200b-3p was significantly upregulated in OSCC, and it was higher in WHO II/III grade than WHO I grade. The AUC of miR-200b-3p for OSCC was 0.9173. miR-200b-3p was significantly downregulated after surgery. High miR-200b-3p expression was associated with poor prognosis.

Discussion and conclusion: Plasma miR-200b-3p could be a potential diagnostic biomarker for OSCC.     

The effect of chronic exposure to extremely low-frequency electromagnetic fields on sleep quality,stress, depression and anxiety

Exposure to extremely low-frequency electromagnetic fields (ELF-EMF) is inevitable in some industries. There are concerns about the possible effects of this exposure. The present study aimed to investigate the effect of chronic exposure to extremely low-frequency electromagnetic fields on sleep quality, stress, depression and anxiety among power plant workers.

In this cross-sectional study, 132 power plant workers were included as the exposed group and 143 other workers were included as the unexposed group. The intensity of ELF-EMF at work stations was measured by using the IEEE Std C95.3.1 standard and then the time weighted average was calculated. Sleep quality, stress, depression and anxiety were measured by using the Pittsburgh Sleep Quality Index Questionnaire; and the Depression, Anxiety and Stress Scale.

The workers in the exposed group experienced significantly poorer sleep quality than the unexposed group. Depression was also more severe in the exposed group than the unexposed group (P = 0.039). Increased exposure to ELF-EMF had a direct and significant relation with increased stress, depression, and anxiety. Sleep quality in technicians with the highest exposure was significantly lower than the other groups.

This study suggests that long-term occupational exposure to ELF-EMF may lead to depression, stress, anxiety and poor sleep quality.     

Gender differences in airway resistance during sleep

Trinder, John, Amanda Kay, Jan Kleiman, and Judith Dunai.Gender differences in airway resistance during sleep.J. Appl. Physiol. 83(6):1986-1997, 1997.At the onset of non-rapid-eye-movement (NREM)sleep there is a fall in ventilation and an increase in upper airwayresistance (UAR). In healthy men there is a progressive increase in UARas NREM sleep deepens. This study compared the pattern of change in UARand ventilation in 14 men and 14 women (aged 18-25 yr) both duringsleep onset and over the NREM phase of a sleep cycle (from wakefulnessto slow-wave sleep). During sleep onset, fluctuations betweenelectroencephalographic alpha and theta activity were associated withmean alterations in inspiratory minute ventilation and UAR of between 1 and 4.5 l/min and between 0.70 and 5.0 cmH₂O · l¹ · s,respectively, with no significant effect of gender on either change(P > 0.05). During NREM sleep,however, the increment in UAR was larger in men than in women(P < 0.01), such that the meanlevels of UAR at peak flow reached during slow-wave sleep were ~25and 10 cmH₂O · l¹ · sin men and women, respectively. We speculate that the greater increasein UAR in healthy young men may represent a gender-related susceptibility to sleep-disordered breathing that, in conjunction withother predisposing factors, may contribute to the development ofobstructive sleep apnea.

     

A novel microRNA miR-1165-3p as a potential diagnostic biomarker for allergic asthma

Context: A further examination of a novel miRNA,miR-1165-3p as a biomarker for asthma, which was previously implicated in helper T cells (Th2) in a murine asthma model.

Objective: To determine whether serum miR-1165-3p can serve as a potential diagnostic biomarker for allergic asthma.

Methods: Serum miR-1165-3p was quantified via quantitative real-time PCR (qRT-PCR) in asthmatic and control samples. Serum miR-1165-3p levels were compared between groups and the clinical diagnostic abilities of miR-1165-3p were evaluated. The analyses utilized included a student’s t test, one-way ANOVA, and the generation of receiver operating characteristic (ROC) curves.

Results: Serum miRNA-1165-3p levels were significantly elevated in asthmatics when compared to the healthy controls. Furthermore, the sensitivity and specificity of serum miR-1165-3p were found to be 83% and 68.2%. Additionally, serum miR-1165-3p levels were also found to be significantly elevated in patients with allergic rhinitis (AR) or allergic bronchopulmonary aspergillosis (ABPA).

Conclusions: This study showed that serum miR-1165-3p can potentially be utilized as a noninvasive biomarker that is able to aid in the diagnosis and characterization of allergic asthma.     

MicroRNAs expression profiles as diagnostic biomarkers of gastric cancer: a systematic literature review

Objective: The early identification of gastric cancer (GC) represents a major clinical challenge. We conducted a systematic review of studies evaluating the miRNA expression profiling as a diagnostic tool in GC.

Methods: We performed a search of PubMed, ISI Web of Science and SCOPUS databases for studies on diagnostic miRNAs and GC, published in English up to October 2017. Eligibility criteria included case-control studies evaluating blood or tissue-based miRNA expression profiles, and incorporating at least two detection phases (screening and validation).

Results: We included 27 eligible studies, that reported on 97 deregulated miRNAs either in blood or tissue, out of which 30 were reported in at least two studies. Among 22 studies on tissue-diagnostic miRNAs, 13 consistently upregulated miRNAs (miR-214, miR-21, miR-103, miR-107, miR-196a, miR-196b, miR-7, miR-135b, miR-222, miR-23b, miR-25, miR-92 and miR-93), and six consistently downregulated miRNAs (miR-148a, miR-375, miR-133b, miR-30a, miR-193a and miR-204) were reported. Ten miRNAs with inconsistent direction of expression in tissues were identified. Among the five studies performed on blood samples, only one miRNA was consistently upregulated (miR-20a).

Conclusions: This review shows that some tissue or blood miRNAs may be considered as potential biomarkers for GC diagnosis, that urgently needs to be confirmed from large prospective studies.     

A probabilistic model for the ultradian timing of REM sleep in mice

A salient feature of mammalian sleep is the alternation between rapid eye movement (REM) and non-REM (NREM) sleep. However, how these two sleep stages influence each other and thereby regulate the timing of REM sleep episodes is still largely unresolved. Here, we developed a statistical model that specifies the relationship between REM and subsequent NREM sleep to quantify how REM sleep affects the following NREM sleep duration and its electrophysiological features in mice. We show that a lognormal mixture model well describes how the preceding REM sleep duration influences the amount of NREM sleep till the next REM sleep episode. The model supports the existence of two different types of sleep cycles: Short cycles form closely interspaced sequences of REM sleep episodes, whereas during long cycles, REM sleep is first followed by an interval of NREM sleep during which transitions to REM sleep are extremely unlikely. This refractory period is characterized by low power in the theta and sigma range of the electroencephalogram (EEG), low spindle rate and frequent microarousals, and its duration proportionally increases with the preceding REM sleep duration. Using our model, we estimated the propensity for REM sleep at the transition from NREM to REM sleep and found that entering REM sleep with higher propensity resulted in longer REM sleep episodes with reduced EEG power. Compared with the light phase, the buildup of REM sleep propensity was slower during the dark phase. Our data-driven modeling approach uncovered basic principles underlying the timing and duration of REM sleep episodes in mice and provides a flexible framework to describe the ultradian regulation of REM sleep in health and disease.     

Altered sleep and behavioral activity phenotypes in PER3-deficient mice

Sleep homeostasis and circadian rhythmicity interact to determine the timing of behavioral activity. Circadian clock genes contribute to circadian rhythmicity centrally and in the periphery, but some also have roles within sleep regulation. The clock gene Period3 (Per3) has a redundant function within the circadian system and is associated with sleep homeostasis in humans. This study investigated the role of PER3 in sleep/wake activity and sleep homeostasis in mice by recording wheel-running activity under baseline conditions in wild-type (WT; n = 54) and in PER3-deficient (Per3(-/-); n = 53) mice, as well as EEG-assessed sleep before and after 6 h of sleep deprivation in WT (n = 7) and Per3(-/-) (n = 8) mice. Whereas total activity and vigilance states did not differ between the genotypes, the temporal distribution of wheel-running activity, vigilance states, and EEG delta activity was affected by genotype. In Per3(-/-) mice, running wheel activity was increased, and REM sleep and NREM sleep were reduced in the middle of the dark phase, and delta activity was enhanced at the end of the dark phase. At the beginning of the baseline light period, there was less wakefulness and more REM and NREM sleep in Per3(-/-) mice. Per3(-/-) mice spent less time in wakefulness and more time in NREM sleep in the light period immediately after sleep deprivation, and REM sleep accumulated more slowly during the recovery dark phase. These data confirm a role for PER3 in sleep-wake timing and sleep homeostasis.     

Effects of night shift on the cognitive load of physicians and urinary steroid hormone profiles – a randomized crossover trial

Mental and physical stress is common in physicians during night shifts. Neurocognitive effects of sleep deprivation as well as alterations in hormonal and metabolic parameters have previously been described. The aim of this crossover study was to evaluate the effects of night-shift work with partial sleep deprivation on steroid hormone excretion and possible associations with mood, sleep characteristics and cognitive functions in physicians.

In total, 34 physicians (mean age 42 ± 8.5 years, 76.5% male) from different departments of the General Hospital of Vienna, Austria, were randomly assigned to two conditions: a regular day shift (8 h on duty, condition 1) and a continuous day-night shift (24 h on duty, condition 2). In both conditions, physicians collected a 24 h urine sample for steroid hormone concentration analysis and further completed psychological tests, including the sleep questionnaire (SF-A), the questionnaire for mental state (MDBF) and the computer-assisted visual memory test (FVW) before and at the end of their shifts, respectively.

Although mean sleep deprivation during night shift was relatively small (~1.5 h) the impairment in participants’ mental state was high in all three dimensions (mood, vigilance and agitation, p ≤ 0.001). Sleep quality (SQ), feeling of being recovered after sleep and mental balance decreased (p ≤ 0.001), whereas mental exhaustion increased (p < 0.05). Moreover, we could show a nearly linear relationship between most of these self-rating items. Testing visual memory participants made significantly more mistakes after night shift (p = 0.011), however, mostly in incorrectly identified items and not in correctly identified ones (FVW). SQ and false identified items were negatively correlated, whereas SQ and time of reaction were positively associated. It is assumed that after night shift, a tendency exists to make faster wrong decisions. SQ did not influence correctly identified items in FVW. In contrast to previous investigations, we found that only excretion rates for pregnanetriol and androsterone/etiocholanolone ratios (p < 0.05, respectively) were slightly reduced in 24-h urine samples after night shift. A considerable stimulation of the adrenocortical axis could not be affirmed. In general, dehydroepiandrosteron (DHEA) was negatively associated with the sense of recreation after sleep and with the time of reaction and positively correlated with correctly identified items in the FVW test.

These results, on the one hand, are in line with previous findings indicating that stress and sleep deprivation suppress gonadal steroids, but, on the other hand, do not imply significant adrenocortical-axis stimulation (e.g. an increase of cortisol) during the day-night shift.     

Sleep and cortical temperature in the Djungarian hamster under baseline conditions and after sleep deprivation

The Djungarian hamster (Phodopus sungorus) is a markedly photoperiodic rodent which exhibits daily torpor under short photoperiod. Normative data were obtained on vigilance states, electroencephalogram (EEG) power spectra (0.25–25.0 Hz), and cortical temperature (T_CRT) under a 168 h light-dark schedule, in 7 Djungarian hamsters for 2 baseline days, 4 h sleep deprivation (SD) and 20 h recovery.During the baseline days total sleep time amounted to 59% of recording time, 67% in the light period and 43% in the dark period. The 4 h SD induced a small increase in the amount of non-rapid eye movement (NREM) sleep and a marked increase in EEG slow-wave activity (SWA; mean power density 0.75–4.0 Hz) within NREM sleep in the first hours of recovery. T_CRT was lower in the light period than in the dark period. It decreased at transitions from either waking or rapid eye movement (REM) sleep to NREM sleep, and increased at the transition from NREM sleep to waking or REM sleep. After SD, T_CRT was lower in all vigilance states.In conclusion, the sleep-wake pattern, EEG spectrum, and time course of T_CRT in the Djungarian hamster are similar to other nocturnal rodents. Also in the Djungarian hamster the time course of SWA seems to reflect a homeostatically regulated process as was formulated in the two-process model of sleep regulation.Abbreviations EEG electroencephalogram - EMG electromyogram - N NREM sleep - NREM non-rapid eye movement - R REM sleep - REM rapid eye movement - SD sleep deprivation - SWA slow-wave activity - T_CRT cortical temperature - TST total sleep time - VS vigilance state - W waking     

Circulating liver-specific microRNAs as noninvasive diagnostic biomarkers of hepatic diseases in human

Context: Hepatitis is an endemic disease worldwide leading to chronic and debilitating cancers. The viral agents and hepatotoxic substances lead to damage of hepatocytes and release of damage associated molecules in circulation. The lack of timely and rapid diagnosis of hepatitis results in chronic disease.

Objective: The present review aimed to describe regulation, release and functions of microRNAs (miR) during human liver pathology and insights into their promising use as noninvasive biomarkers of hepatitis.

Methods: Comprehensive data were collected from PubMed, ScienceDirect and the Web of Science databases utilizing the keywords “biomarkers”, “microRNAs” and “hepatic diseases”.

Results: The miRs are readily released in the body fluids and blood during HBV/HCV associated hepatitis as well as metabolic, alcoholic, drug induced and autoimmune hepatitis. The liver-specific microRNAs including miR-122, miR-130, miR-183, miR-196, miR-209 and miR-96 are potential indicators of liver injury (mainly via apoptosis, necrosis and necroptosis) or hepatitis with their varied expression during acute/fulminant, chronic, liver fibrosis/cirrhosis and hepato-cellular carcinoma.

Conclusions: The liver-specific miRs can be used as rapid and noninvasive biomarkers of hepatitis to discern different stages of hepatitis. Blocking or stimulating pathways associated with miR regulation in liver could unveil novel therapeutic strategies in the management of liver diseases.

• Clinical significance

• Liver specific microRNAs interact with cellular proteins and signaling molecules to regulate the expression of various genes controlling biological processes.

• The circulatory level of liver specific microRNAs is indicator of severity of HBV and HCV infections as well as prognostic and therapeutic candidates.

• The expression of liver specific microRNAs is strongly associated with infectious, drug-induced, hepatotoxic, nonalcoholic steatohepatitis and nonalcoholic fatty liver diseases.