Component Of Circadian Rhythm Research Articles

Analysis of circadian rhythm components in EEG/EMG data of aged mice.

Aging disrupts circadian clocks, as evidenced by a reduction in the amplitude of circadian rhythms. Because the circadian clock strongly influences sleep-wake behavior in mammals, age-related alterations in sleep-wake patterns may be attributable, at least partly, to functional changes in the circadian clock. However, the effect of aging on the circadian characteristics of sleep architecture has not been well assessed, as circadian behaviors are usually evaluated through long-term behavioral recording with wheel-running or infrared sensors. In this study, we examined age-related changes in circadian sleep-wake behavior using circadian components extracted from electroencephalography (EEG) and electromyography (EMG) data. EEG and EMG were recorded from 12 to 17-week-old and 78 to 83-week-old mice for 3 days under light/dark and constant dark conditions. We analyzed time-dependent changes in the duration of sleep. Rapid eye movement (REM) and non-REM (NREM) sleep significantly increased during the night phase in old mice, whereas no significant change was observed during the light phase. The circadian components were then extracted from the EEG data for each sleep-wake stage, revealing that the circadian rhythm in the power of delta waves during NREM sleep was attenuated and delayed in old mice. Furthermore, we used machine learning to evaluate the phase of the circadian rhythm, with EEG data serving as the input and the phase of the sleep-wake rhythm (environmental time) as the output. The results indicated that the output time for the old mice data tended to be delayed, specifically at night. These results indicate that the aging process significantly impacts the circadian rhythm in the EEG power spectrum despite the circadian rhythm in the amounts of sleep and wake attenuated but still remaining in old mice. Moreover, EEG/EMG analysis is useful not only for evaluating sleep-wake stages but also for circadian rhythms in the brain.

Circadian regulation of mTORC1 signaling via Per2 dependent mechanism disrupts folliculogenesis and oocyte maturation in female mice.

mTOR (mammalian target of Rapamycin) is an important signaling pathway involved in several crucial ovarian functions including folliculogenesis and oocyte maturation. The circadian rhythm regulates multiple physiological processes and PER2 is one of the core circadian rhythm components. mTOR is regulated by the circadian clock and in turn, the rhythmic mTOR activities strengthen the clock function. Our current study aims to investigate a possible interconnection between the circadian clock and the mTORC1 signaling pathway in folliculogenesis and oocyte maturation. Here we demonstrate that the circadian system regulates mTORC1 signaling via Per2 dependent mechanism in the mouse ovary. To investigate the effect of constant light on ovarian and oocyte morphology, animals were housed 12:12h L:D group in standard lightening conditions and the 12:12h L:L group in constant light for one week. Food intake and body weight changes were measured. Ovarian morphology, follicle counting, and oocyte aging were evaluated. Afterward, western blot for mTOR, p-mTOR, p70S6K, p-p70S6K, PER2, and Caspase-3 protein levels was performed. The study demonstrated that circadian rhythm disruption caused an alteration in their food intake and decrease in primordial follicle numbers and an increase in the number of atretic follicles. It caused an increase in oxidative stress and a decrease in ZP3 expression in oocytes. Decreased protein levels of mTOR, p-mTOR, p70S6K, and PER2 were shown. The results showed that the circadian clock regulates mTORC1 through PER2 dependent mechanism and that decreased mTORC1 activity can contribute to premature aging of mouse ovary. In conclusion, these results suggest that the circadian clock may control ovarian aging by regulating mTOR signaling pathway through Per2 expression.

Melatonin as a Harmonizing Factor of Circadian Rhythms, Neuronal Cell Cycle and Neurogenesis: Additional Arguments for Its Therapeutic Use in Alzheimer's Disease.

The synthesis and release of melatonin in the brain harmonize various physiological functions. The apparent decline in melatonin levels with advanced aging is an aperture to the neurodegenerative processes. It has been indicated that down regulation of melatonin leads to alterations of circadian rhythm components, which further causes a desynchronization of several genes and results in an increased susceptibility to develop neurodegenerative diseases. Additionally, as circadian rhythms and memory are intertwined, such rhythmic disturbances influence memory formation and recall. Besides, cell cycle events exhibit a remarkable oscillatory system, which is downstream of the circadian phenomena. The linkage between the molecular machinery of the cell cycle and complex fundamental regulatory proteins emphasizes the conjectural regulatory role of cell cycle components in neurodegenerative disorders such as Alzheimer's disease. Among the mechanisms intervening long before the signs of the disease appear, the disturbances of the circadian cycle, as well as the alteration of the machinery of the cell cycle and impaired neurogenesis, must hold our interest. Therefore, in the present review, we propose to discuss the underlying mechanisms of action of melatonin in regulating the circadian rhythm, cell cycle components and adult neurogenesis in the context of AD pathogenesis with the view that it might further assist to identify new therapeutic targets.

Integrative multi-omics analysis of three early diverged rosid species reveals an ancient hierarchical cold-responsive regulatory network.

Elucidating regulators, including transcription factors (TFs) and RNA-binding proteins (RBPs), underlying gene transcriptional and post-transcriptional co-regulatory network is key to understand plant cold responses. Previous studies were mainly conducted on single species, and whether the regulators are conserved across different species remains elusive. Here, we selected three species that diverged at the early evolution of rosids (~99-113 million years ago), performed cold-responsive phylotranscriptome experiments, and integrated chromatin immunoprecipitation- and DNA affinity purification-sequencing (ChIP/DAP-seq) analysis to explore cold-responsive regulators and their regulatory networks. First, we detected over 10,000 cold-induced differentially expressed genes (DEGs) and alternative splicing genes (DASGs) in each species. Among the DEGs, a set of TFs and RBPs were conserved in rosid cold response. Compared to TFs, RBPs displayed a delayed cold-responsive pattern, implying a hierarchical regulation of DEGs and DASGs. By integrating DEGs and DASGs, we identified 259 overlapping DE-DASG orthogroups (closely-related homologs) that were cold-regulated at both transcriptional and post-transcriptional levels in all three studied species. Notably, pathway analysis on each of the DEGs, DASGs, and DE-DASGs in the three species showed a common enrichment connected to the circadian rhythm. Evidently, 226 cold-responsive genes were directly targeted by at least two circadian rhythm components (CCA1, LHY, RV4, RVE7, and RVE8). Finally, we revealed an ancient hierarchy of cold-responsive regulatory networks at transcriptional and post-transcriptional levels launched by circadian components in rosids. Altogether, this study sheds light on conserved regulators underlying cold-responsive regulatory networks across rosid species, despite a long evolutionary history after their divergence.

Circadian rhythms and cancer.

Circadian rhythms are autonomous oscillators developed by the molecular circadian clock, essential for coordinating internal time with the external environment in a 24-h daily cycle. In mammals, this circadian clock system plays a major role in all physiological processes and severely affects human health. The regulation of the circadian clock extends beyond the clock genes to involve several clock-controlled genes. Hence, the aberrant expression of these clock genes leads to the downregulation of important targets that control the cell cycle and the ability to undergo apoptosis. This may lead to genomic instability and promotes carcinogenesis. Alteration in the clock genes and their modulation is recognized as a new approach for the development of effective treatment against several diseases, including cancer. Until now, there has been a lack of understanding of circadian rhythms and cancer disease. For that, this chapter aims to represent the core components of circadian rhythms and their function in cancer pathogenesis and progression. In addition, the clinical impacts, current clock drugs, and potential therapeutic targets have been discussed.

Modulatory roles of melatonin on respiratory and heart rates and their circadian rhythmicity in donkeys (Equus asinus) subjected to packing during the hot-dry season.

In mammals a central circadian clock is located in the suprachiasmatic nuclei of the hypothalamus, which regulates the innate physiological rhythms to the ambient 24-h light-dark cycle. Melatonin is an essential component of circadian rhythm. The study aimed to evaluate the effects of melatonin administration on the respiratory rate (RR) and heart rate (HR) and their circadian rhythmicity in donkeys subjected to packing (load carrying) during the hot-dry season. Twenty healthy pack donkeys, aged 2–3 years with average weight of 93 ± 2.7 kg were divided into two groups randomly for the experiment. Group 1 donkeys (packing with melatonin) were administered melatonin for seven days during the study and subjected to packing, while group 2 were packed without melatonin administration. Both groups of donkeys were packed three times within the week, one day apart, covering a total distance of 20 km on each experimental day. Meteorological parameters were recorded during the study period. RR and HR were measured pre- and immediately (15 min) post-packing. Continuous measurement of the parameters started 16 h later, after the last packing procedures for a period of 27 h at intervals of 3 h. Temperature-humidity index (THI) pre-packing (73.67 ± 0.7) was lower (P < 0.05) than that obtained post-packing (80.33 ± 1.2). The THI recorded during the continuous measurement was at its peak at 15:00 h (86), indicating that the afternoon hours were thermally stressful to the donkeys. The THI was strongly correlated with HR recorded in packing (without melatonin) compared to packing (with melatonin) donkeys. The RR and HR values in both groups of donkeys were significantly (P < 0.05) higher immediately after packing. However, the post-packing values of the parameters were not different (P > 0.05) between the two groups of donkeys. The mesor (adjusted arithmetic mean) and amplitude of RR and HR in packing (without melatonin) donkeys were significantly (P < 0.05) higher than that recorded in packing (with melatonin) donkeys. In conclusion, melatonin reduced negative influence on the circadian rhythmicity (mesor and amplitude) of RR and HR in donkeys by exerting its anti-stress and antioxidant effects. The study has demonstrated the beneficial effect of melatonin and its administration may mitigate excessive respiratory and cardiac activities that may reduce the work output of donkeys during the hot-dry season.

Down-regulation of BMAL1 by MiR-494-3p Promotes Hepatocellular Carcinoma Growth and Metastasis by Increasing GPAM-mediated Lipid Biosynthesis.

The circadian clock confers daily rhythmicity to many crucial biological processes and behaviors and its disruption is closely associated with carcinogenesis in several types of cancer. Brain and muscle arnt-like protein 1 (BMAL1) is a core circadian rhythm component in mammals and its dysregulation has been shown to contribute to aberrant metabolism in human diseases. However, the biological functions of BMAL1, especially its involvement in aberrant lipid metabolism in hepatocellular carcinoma (HCC), remain elusive. In the present study, we found that BMAL1 was frequently down-regulated in HCC cells mainly due to the up-regulation of miR-494-3p. Down-regulation of BMAL1 was significantly associated with poor survival in HCC patients. BMAL1 down-regulation promoted HCC cell growth and metastasis both in vitro and in vivo. Mechanistically, through cooperating with EZH2, BMAL1 transcriptionally suppressed the expression of glycerol-3-phosphate acyltransferase mitochondrial (GPAM), a key enzyme involved in the regulation of lipid biosynthesis, leading to reduced levels lysophosphatidic acid (LPA), which have long been known as mediator of oncogenesis. Particularly, treatment with SR8278, an activator of BMAL1, exhibited a therapeutic effect on HCC in vitro and in vivo. In conclusion, BMAL1 plays a critical anti-oncogenic role in HCC, providing strong research evidence for BMAL1 as a prospective target for HCC therapy.

Circadian Dysregulation: A Mainstream Mess of the Present World

The physiological systems of humans and other organisms are periodic in nature. One such system is a circadian rhythm, a biological internal clock that is endogenous and entrainable. The circadian rhythm regulates essential functions such as the sleep/wake cycle, hormones, feeding behaviour, metabolism and cell division as well. Due to shift work or jet lag or even irregular sleep, diet, etc., circadian rhythm disorders are one of the most common problems in this century. It is a major factor that can trigger various diseases like depression, lung tumorigenesis, cancer, anxiety, depression and many more. The purpose of this review is to discuss circadian dysregulation and its potential long-term effects in cancer including lung tumor and mental illness including depression, anxiety. Loss of autonomous cells containing Bmal1 and Per2 (the core components of circadian rhythm) will increase growth and metabolism imbalance and increase in c-Myc levels. To treat circadian rhythm disorders, zeitgebers (external cues) should be used to entrain or synchronize the circadian rhythm and sleep phase chronotherapy can also be used.

O-158 The effect of circadian rhythm disruption due to constant light on ovarian and oocyte aging through possible relationship between PER2 and mTOR signaling pathway

Abstract Study question Does circadian rhythm disruption by constant light affect the ovarian morphology and function, and cause ovarian and oocyte aging through possible relationship between PER2 and mTOR? Summary answer We demonstrated that circadian rhythm disruption by light may cause ovarian and oocyte aging. What is known already Circadian rhythm regulates multiple physiological processes and PER2 is one of the core circadian rhythm components. Changes in light conditions may cause circadian rhythm disruptions. Light exposure at night may cause attenuation in PER2 mRNA and protein levels. Circadian rhythm disruptions are thought to be associated with reproductive diseases. mTOR signaling pathway functions in folliculogenesis and oocyte maturation in ovary. Also, it is associated with ovarian and oocyte aging. Study design, size, duration A total of 32 female Balb/c mice which enter estrous cycle were used in the study. Mice were randomly assigned to one of two groups as 12:12h L:D and 12:12h L:L. During the experiment, 12:12h L:D (control group) was housed in a 12:12h light:dark cycle and 12:12h L:L (experiment group) was housed in a constant light conditions 12:12h light:light for 1 week. Participants/materials, setting, methods We housed 12:12h L:D group in standard lightening conditions and 12:12h L:L group in constant light for one week. We performed food intake and body weight change analysis. We evaluated ovarian morphology, follicle counting analysis. We evaluated ZP3 and nitrotyrosine (NTY) expression for oocyte aging markers. We performed western blot for PER2, mTOR, p-mTOR, p70 S6K, p-p70 S6K, and Caspase-3 protein levels. Main results and the role of chance We demonstrated that circadian rhythm disruption caused alteration in their food intake and decrease in primordial follicle numbers and increase in atretic follicles (p &amp;lt; 0.05). It caused increase in oxidative stress and decrease in ZP3 expression in oocytes (p &amp;lt; 0.05). We showed decreased protein levels of PER2, mTOR, p-mTOR and p70 S6K (p &amp;lt; 0.05). Limitations, reasons for caution The explanation of molecular mechanism underlying the relationship between circadian rhythm disruptions by light and ovarian function may lead the usage of circadian rhythm-based or light-based therapies currently using to treat some diseases on female reproductive system related diseases. Wider implications of the findings We conclude that constant light may reduce follicle reserve, cause follicles to go rapidly atresia and disrupt the oocyte quality, thus it may be a risk factor for female reproductive diseases such as premature ovarian insufficiency and early menopause. Trial registration number not applicable

Dual function of clock component OsLHY sets critical day length for photoperiodic flowering in rice.

SummaryCircadian clock, an endogenous time‐setting mechanism, allows plants to adapt to unstable photoperiod conditions and induces flowering with proper timing. In Arabidopsis, the central clock oscillator was formed by a series of interlocked transcriptional feedback loops, but little is known in rice so far. By MutMap technique, we identified the candidate gene OsLHY from a later flowering mutant lem1 and further confirmed it through genetic complementation, RNA interference knockdown, and CRISPR/Cas9‐knockout. Global transcriptome profiling and expression analyses revealed that OsLHY might be a vital circadian rhythm component. Interestingly, oslhy flowered later under ≥12 h day length but headed earlier under ≤11 h day length. qRT‐PCR results exhibited that OsLHY might function through OsGI‐Hd1 pathway. Subsequent one‐hybrid assays in yeast, DNA affinity purification qPCR, and electrophoretic mobility shift assays confirmed OsLHY could directly bind to the CBS element in OsGI promoter. Moreover, the critical day length (CDL) for function reversal of OsLHY in oslhy (11–12 h) was prolonged in the double mutant oslhy osgi (about 13.5 h), indicating that the CDL set by OsLHY was OsGI dependent. Additionally, the dual function of OsLHY entirely relied on Hd1, as the double mutant oslhy hd1 showed the same heading date with hd1 under about 11.5, 13.5, and 14 h day lengths. Together, OsLHY could fine‐tune the CDL by directly regulating OsGI, and Hd1 acts as the final effector of CDL downstream of OsLHY. Our study illustrates a new regulatory mechanism between the circadian clock and photoperiodic flowering.

Circadian Rhythms in Legumes: What Do We Know and What Else Should We Explore?

The natural timing devices of organisms, commonly known as biological clocks, are composed of specific complex folding molecules that interact to regulate the circadian rhythms. Circadian rhythms, the changes or processes that follow a 24-h light–dark cycle, while endogenously programmed, are also influenced by environmental factors, especially in sessile organisms such as plants, which can impact ecosystems and crop productivity. Current knowledge of plant clocks emanates primarily from research on Arabidopsis, which identified the main components of the circadian gene regulation network. Nonetheless, there remain critical knowledge gaps related to the molecular components of circadian rhythms in important crop groups, including the nitrogen-fixing legumes. Additionally, little is known about the synergies and trade-offs between environmental factors and circadian rhythm regulation, especially how these interactions fine-tune the physiological adaptations of the current and future crops in a rapidly changing world. This review highlights what is known so far about the circadian rhythms in legumes, which include major as well as potential future pulse crops that are packed with nutrients, particularly protein. Based on existing literature, this review also identifies the knowledge gaps that should be addressed to build a sustainable food future with the reputed “poor man’s meat”.

Effects of prolonged night-time light exposure and traffic noise on the behavior and body temperature rhythmicity of the wild desert rodent, Gerbillus tarabuli

ABSTRACT The aim of this study was to demonstrate for the first time in Tarabul’s gerbils (Gerbillus tarabuli), the effects of simultaneous exposure to two major environmental stressors – light and noise pollutions – on the body temperature rhythm and anxious behavior. Seven groups, each consisting of 6 adult male gerbils, were subjected to a standard LD cycle (12 L:12D) with lights on at 08:00 h and off at 20:00 h, constant conditions (total darkness, DD), prolonged nighttime exposure to light (PEL: 18 L:6D) with lights on at 08:00 h and off at 02:00 h, mimicking prolonged exposure to light pollution in peri-urban areas, exposure to auditory stress (TNS) of 80 dB, and conditions combining PEL&TNS. The body temperature circadian rhythm was recorded, and behavioral tests were performed at the end of experimental phases. The results revealed the existence, for the first time in Gerbilus tarabuli, of an endogenous circadian rhythm of body temperature with a period of 23.8 ± 0.04 h. Prolonged exposure to light at night (PEL) induced a significant phase delay (02 h 09 min ± 0.16 h) of the rhythm, with an acrophase (peak time) occurring at 04:42 ± 0.13 h instead of 02:33 ± 0.21 h. Exposure to TNS for 4 hours per night induced a significant increase of the amplitude of the rhythm and a decrease of the rhythm regularity (robustness of 73.26% in TNS vs. 82.32 in control condition). While combining TNS and PEL significantly delayed the phase of the Tb rhythm by 3 h 10 min (acrophase at 06:39 ± 0.37 h instead of 02:33 ± 0.21 h), increased the amplitude, and significantly reduced the stability of the rhythm (robustness of 67.25% in PEL&TNS vs. 82.32 in control condition). PEL&TNS and TNS environments induce an important stress in gerbils highlighted by a significant decrease of the number of line crossings and time spent in the center area of the open field test. Furthermore, elevated plus maze test revealed gerbils of the PEL&TNS and TNS conditions significantly visited the lowest number of open arms and spent a shorter amount of time in it. In addition, these conditions were responsible for less activity (total number of entries in arms) than in the control and PEL conditions. These results indicate clearly that in the desert area, peri-urban light and noise pollutions disturb the circadian rhythm components and alter the behavior of Tarabul’s gerbils inducing an anxious state.

Bmal1 Deletion in Myeloid Cells Attenuates Atherosclerotic Lesion Development and Restrains Abdominal Aortic Aneurysm Formation in Hyperlipidemic Mice.

Although the molecular components of circadian rhythms oscillate in discrete cellular components of the vasculature and many aspects of vascular function display diurnal variation, the cellular connections between the molecular clock and inflammatory cardiovascular diseases remain to be elucidated. Previously we have shown that pre- versus postnatal deletion of Bmal1 (brain and muscle aryl hydrocarbon receptor nuclear translocator-like 1), the nonredundant core clock gene has contrasting effects on atherogenesis. Here we investigated the effect of myeloid cell Bmal1 deletion on atherogenesis and abdominal aortic aneurysm formation in mice. Approach and Results: Mice lacking Bmal1 in myeloid cells were generated by crossing Bmal1 flox/flox mice with lysozyme 2 promoter-driven Cre recombinase mice on a hyperlipidemic low-density lipoprotein receptor-deficient background and were fed on a high-fat diet to induce atherosclerosis. Atherogenesis was restrained, concomitant with a reduction of aortic proinflammatory gene expression in myeloid cell Bmal1 knockout mice. Body weight, blood pressure, blood glucose, triglycerides, and cholesterol were unaltered. Similarly, myeloid cell depletion of Bmal1 also restrained Ang II (angiotensin II) induced formation of abdominal aortic aneurysm in hyperlipidemic mice. In vitro, RNA-Seq analysis demonstrated a proinflammatory response in cultured macrophages in which there was overexpression of Bmal1. Myeloid cell Bmal1 deletion retards atherogenesis and restrains the formation of abdominal aortic aneurysm and may represent a potential therapeutic target for inflammatory cardiovascular diseases.

PK/PD modeling of 5-hydroxytryptophan (5-HTP) challenge test with cortisol measurement in serum and saliva.

This research was planned to build a Pharmacokinetic/Pharmacodynamic (PK/PD) model of 5‐hydroxytryptophan (5‐HTP) challenge study including a circadian rhythm component of cortisol and to predict serum cortisol based on saliva cortisol. Data from three 5‐HTP challenge studies in healthy volunteers were collected. Serum 5‐HTP, saliva, and serum cortisol were sampled as PK and PD marker. The population PK/PD modeling approach was applied. A baseline model of serum cortisol was built to assess the circadian rhythm before a pharmacodynamic model was used to evaluate the drug effect of the 5‐HTP on cortisol. Finally, linear and power function relationships were tested to predict serum cortisol based on saliva cortisol. The PK of 5‐HTP could be described using a one‐compartment model with a transit compartment. The typical value for clearance was 20.40 L h−1 and showed inter‐study variability. A cosine function was chosen and properly described the circadian rhythm of serum cortisol. A linear approximation model was applied to fit the 5‐HTP PD effect on cortisol data with a slope of 4.16 ng mL−1 h. A power function provided a better description than a linear function to relate the saliva and serum cortisol. In conclusion, a circadian rhythm component was built in the PK/PD model of the 5‐HTP challenge test which could better improve the understanding of the stimulating effect on HPA with cortisol change. After the 5‐HTP challenge, saliva cortisol correlated well with serum cortisol and was predictable by a population PK‐PD model.

Cryptochromes modulate E2F family transcription factors

Early 2 factor (E2F) family transcription factors participate in myriad cell biological processes including: the cell cycle, DNA repair, apoptosis, development, differentiation, and metabolism. Circadian rhythms influence many of these phenomena. Here we find that a mammalian circadian rhythm component, Cryptochrome 2 (CRY2), regulates E2F family members. Furthermore, CRY1 and CRY2 cooperate with the E3 ligase complex SKP-CULLIN-FBXL3 (SCFFBXL3) to reduce E2F steady state protein levels. These findings reveal an unrecognized molecular connection between circadian clocks and cell cycle regulation and highlight another mechanism to maintain appropriate E2F protein levels for proper cell growth.

A computationally efficient algorithm to obtain an accurate and interpretable model of the effect of circadian rhythm on resting heart rate

Objective: Wrist-worn wearable devices equipped with heart rate (HR) sensors have become increasingly popular. The ability to correctly interpret the collected data is fundamental to analyse user’s well-being and perform early detection of abnormal physiological data. Circadian rhythm is a strong factor of variability in HR, yet few models attempt to accurately model its effect on HR. Approach: In this paper we present a mathematical derivation of the single-component cosinor model with multiple components that fits user data to a predetermined arbitrary function (the expected shape of the circadian effect on resting HR (RHR)), thus permitting us to predict the user’s circadian rhythm component (i.e. MESOR, Acrophase and Amplitude) with a high accuracy. Main results: We show that our model improves the accuracy of HR prediction compared to the single component cosinor model (10% lower RMSE), while retaining the readability of the fitted model of the single component cosinor. We also show that the model parameters can be used to detect sleep disruption in a qualitative experiment. The model is computationally cheap, depending linearly on the size of the data. The computation of the model does not need the full dataset, but only two surrogates, where the data is accumulated. This implies that the model can be implemented in a streaming approach, with important consequences for security and privacy of the data, that never leaves the user devices. Significance: The multiple component model provided in this paper can be used to approximate a user’s RHR with higher accuracy than single component model, providing traditional parameters easy to interpret (i.e. the same produced by the single component cosinor model). The model we developed goes beyond fitting circadian activity on RHR, and it can be used to fit arbitrary periodic real valued time series, vectorial data, or complex data.

Saccadic eye movements estimate prolonged time awake.

Prolonged time awake increases the need to sleep. Sleep pressure increases sleepiness, impairs human alertness and performance and increases the probability of human errors and accidents. Human performance and alertness during waking hours are influenced by homeostatic sleep drive and the circadian rhythm. Cognitive functions, especially attentional ones, are vulnerable to circadian rhythm and increasing sleep drive. A reliable, objective and practical metrics for estimating sleepiness could therefore be valuable. Our aim is to study whether saccades measured with electro-oculography (EOG) outside the laboratory could be used to estimate the overall time awake without sleep of a person. The number of executed saccades was measured in 11 participants during an 8-min saccade task. The saccades were recorded outside the laboratory (Naval Academy, Bergen) using EOG every sixth hour until 54hr of time awake. Measurements were carried out on two occasions separated by 10weeks. Five participants participated in both measurement weeks. The number of saccades decreased during sustained wakefulness. The data correlated with the three-process model of alertness; performance differed between participants but was stable within individual participants. A mathematically monotonous relation between performance in the saccade task and time awake was seen after removing the circadian rhythm component from measured eye movement data. The results imply that saccades measured with EOG can be used as a time-awake metric outside the laboratory.

Diurnal changes in photosynthesis by six submerged macrophytes measured using fluorescence

The diurnal changes of chlorophyll fluorescence of terrestrial plants have been well studied and show a midday depression in summer. However, only a few studies have been performed on submerged macrophytes though they play important roles in shallow lakes. In the present study, six common submerged macrophytes were chosen to investigate the diurnal changes of the maximum potential quantum efficiency of photosystem II (Fv/Fm) on sunny days in summer. The Fv/Fm values of all these species decreased at midday under natural conditions, but there was no significant change either under the “Shade” treatment or under laboratory conditions. The Fv/Fm values were negatively correlated with photon radiance, except for Ceratophyllum demersum, which probably could support high light as shown by rapid light curves. The decreasing Fv/Fm values during midday known in terrestrial plants to be caused by either down-regulation or photodamage of photosystem II as a response to high irradiances was here also demonstrated for submerged freshwater macrophytes. In Vallisneria denserrulata showed an additional component of circadian rhythm to the midday photoinhibition. Circadian periodicity and the diurnal changes of the photon irradiance should be considered when investigating the chlorophyll fluorescence of submerged macrophytes.

Natural variation in CIRCADIAN CLOCK ASSOCIATED 1 is associated with flowering time in Brassica rapa.

Flowering time is a very important agronomic trait and the development of molecular markers associated with this trait can facilitate crop breeding. CIRCADIAN CLOCK ASSOCIATED 1 (CCA1), a core oscillator component of circadian rhythms that affect metabolic pathways in plants, has been implicated in flowering time control in species of Brassica. CCA1 gene sequences from three Brassica rapa inbred lines, showing either early flowering or late flowering phenotypes, were analyzed and a high level of sequence variation was identified, especially within the fourth intron. Using this information, three PCR primer sets were designed and tested using various inbred lines of B. rapa. The usage of InDel markers was further validated by evaluation of flowering time and high resolution melting (HRM) analysis. Both methods, PCR and HRM, validated the use of newly developed markers. Additional sequence analyses of Brassica plants with diploid (AA, BB, or CC) and allotetraploid genomes further confirmed a large number of sequence polymorphisms in the CCA1 gene, including insertions/deletions in the fourth intron. Our results demonstrated that sequence variations in CCA1 can be used to develop valuable trait-related molecular markers for Brassica crop breeding.

The role of casein kinase I in the Drosophila circadian clock.

The circadian clock mechanism in organisms as diverse as cyanobacteria and humans involves both transcriptional and posttranslational regulation of key clock components. One of the roles for the posttranslational regulation is to time the degradation of the targeted clock proteins, so that their oscillation profiles are out of phase with respect to those of the mRNAs from which they are translated. In Drosophila, the circadian transcriptional regulator PERIOD (PER) is targeted for degradation by a kinase (DOUBLETIME or DBT) orthologous to mammalian kinases (CKIɛ and CKIδ) that also target mammalian PER. Since these kinases are not regulated by second messengers, the mechanism (if any) for their regulation is not known. We are investigating the possibility that regulation of DBT is conferred by other proteins that associate with DBT and PER. In this chapter, the methods we are employing to identify and analyze these factors are discussed. These methods include expression of wild type and mutant proteins with the GAL4/UAS binary expression approach, analysis of DBT in Drosophila S2 cells, in vitro kinase assays with DBT isolated from S2 cells, and proteomic analysis of DBT-containing complexes and of DBT phosphorylation with mass spectrometry. The work has led to the discovery of a previously unrecognized circadian rhythm component (Bride of DBT, a noncanonical FK506-binding protein) and the mapping of autophosphorylation sites within the DBT C-terminal domain with potential regulatory roles.