Circadian Phase Shifts Research Articles

Human phase response curve to a 1 h pulse of bright white light

The phase resetting response of the human circadian pacemaker to light depends on the timing of exposure and is described by a phase response curve (PRC). The current study aimed to construct a PRC for a 1 h exposure to bright white light (∼8000 lux) and to compare this PRC to a <3 lux dim background light PRC. These data were also compared to a previously completed 6.7 h bright white light PRC and a <15 lux dim background light PRC constructed under similar conditions. Participants were randomized for exposure to 1 h of either bright white light (n=18) or <3 lux dim background light (n=18) scheduled at 1 of 18 circadian phases. Participants completed constant routine (CR) procedures in dim light (<3 lux) before and after the light exposure to assess circadian phase. Phase shifts were calculated as the difference in timing of dim light melatonin onset (DLMO) during pre- and post-stimulus CRs. Exposure to 1 h of bright white light induced a Type 1 PRC with a fitted peak-to-trough amplitude of 2.20 h. No discernible PRC was observed in the <3 lux dim background light PRC. The fitted peak-to-trough amplitude of the 1 h bright light PRC was ∼40% of that for the 6.7 h PRC despite representing only 15% of the light exposure duration, consistent with previous studies showing a non-linear duration–response function for the effects of light on circadian resetting.

Effects of partial circadian adjustments on sleep and vigilance quality during simulated night work

In most situations, complete circadian adjustment is not recommended for night workers. With complete adjustment, workers experience circadian misalignment when returning on a day-active schedule, causing repeated circadian phase shifts and internal desynchrony. For this reason, partial circadian realignment was proposed as a good compromise to stabilize internal circadian rhythms in night shift workers. However, the extent of partial circadian adjustment necessary to improve sleep and vigilance quality is still a matter of debate. In this study, the effects of small but statistically significant partial circadian adjustments on sleep and vigilance quality were assessed in a laboratory simulation of night work to determine whether they were also of clinical significance. Partial adjustments obtained by phase delay or by phase advance were quantified not only by the phase shift of dim light salivary melatonin onset, but also by the overlap of the episode of melatonin production with the sleep-wake cycle adopted during simulated night work. The effects on daytime sleep and night-time vigilance quality were modest. However, they suggest that even small adjustments by phase delay may decrease the accumulation of sleep debt, whereas the advance strategy improves subjective alertness and mood during night work. Furthermore, absolute phase shifts, by advance or by delay, were associated with improved subjective alertness and mood during the night shift. These strategies need to be tested in the field, to determine whether they can be adapted to real-life situations and provide effective support to night workers.

Downregulation of Clock in circulatory system leads to an enhancement of fibrinolysis in mice

As a main component of circadian genes, clock plays not only an important role in circadian rhythm but also in the regulation of many physiological systems. The dysfunction of clock genes is associated with the development of various disorders. Many studies have investigated the association between clock genes and blood coagulation and the fibrinolytic system. The present study was designed to investigate the effect of downregulation of circulatory Clock on blood coagulation and fibrinolysis at the initial stage of active phase in male mice. Downregulation of the expression of the Clock gene by siRNA and, subsequently, its effect on the thrombotic potential and the expression of relative coagulative and/or fibrinolytic factors were investigated. It was found that the Clock interfered mice were less liable to thrombosis and showed prolonged prothrombin time (PT) and activated partial thromboplastin time (APTT) at Zeitgeber time (ZT) 15. Meanwhile, these mice also showed an increase in factor VII (FVII) and a decrease in thrombomodulin (TM) and plasminogen activator inhibitor 1 (PAI-1) at ZT 15 at both transcriptional and translational levels. PT, APTT and mRNA expressions of fvii, tm and pai-1 were analyzed with the least-squares fit of a 24-h cosine function by single cosinor method; no circadian rhythm was determined in PT and APTT, and a higher amplitude of fvii in the Clock RNAi group was found with a circadian phase shift, while lower amplitudes of tm and pai-1 were found in the Clock RNAi group with nearly no phase shift. All these results suggest that downregulation of the Clock gene in circulatory system has an effect on factors involved in both blood coagulation and fibrinolysis resulting in an enhancement in mice. This may be considered as an indication that Clock regulates thrombotic homeostasis through the fibrinolytic system.

Loss of dexras1 Alters Nonphotic Circadian Phase Shifts and Reveals a Role for the Intergeniculate Leaflet (IGL) in Gene-Targeted Mice

Loss of Dexras1 in gene-targeted mice impairs circadian entrainment to light cycles and produces complex changes to phase-dependent resetting responses (phase shifts) to light. The authors now describe greatly enhanced and phase-specific nonphotic responses induced by arousal in dexras1−/− mice. In constant conditions, mutant mice exhibited significant arousal-induced phase shifts throughout the subjective day. Unusual phase advances in the late subjective night were also produced when arousal has little effect in mice. Bilateral lesions of the intergeniculate leaflet (IGL) completely eliminated both the nonphotic as well as the light-induced phase shifts of circadian locomotor rhythms during the subjective day, but had no effect on nighttime phase shifts. The expression of FOS-like protein in the suprachiasmatic nucleus (SCN) was not affected by either photic or nonphotic stimulation in the subjective day in either genotype. Therefore, the loss of Dexras1 (1) enhances nonphotic phase shifts in a phase-dependent manner, and (2) demonstrates that the IGL in mice is a primary mediator of circadian phase-resetting responses to both photic and nonphotic events during the subjective day, but plays a different functional role in the subjective night. Furthermore, (3) the change in FOS level does not appear to be a critical step in the entrainment pathways for either light or arousal during the subjective day. The cumulative evidence suggests that Dexras1 regulates multiple photic and nonphotic signal-transduction pathways, thereby playing an essential role modulating species-specific characteristics of circadian entrainment. (Author correspondence: ralph@psych.utoronto.ca)

Effects of an advanced sleep schedule and morning short wavelength light exposure on circadian phase in young adults with late sleep schedules

We examined the effects of an advanced sleep/wake schedule and morning short wavelength (blue) light in 25 adults (mean age±SD=21.8±3 years; 13 women) with late sleep schedules and subclinical features of delayed sleep phase disorder (DSPD). After a baseline week, participants kept individualized, fixed, advanced 7.5-h sleep schedules for 6days. Participants were randomly assigned to groups to receive "blue" (470nm, ∼225lux, n=12) or "dim" (<1lux, n=13) light for 1h after waking each day. Head-worn "Daysimeters" measured light exposure; actigraphs and sleep diaries confirmed schedule compliance. Salivary dim light melatonin onset (DLMO), self-reported sleep, and mood were examined with 2×2 ANOVA. After 6days, both groups showed significant circadian phase advances, but morning blue light was not associated with larger phase shifts than dim-light exposure. The average DLMO advances (mean±SD) were 1.5±1.1h in the dim light group and 1.4±0.7h in the blue light group. Adherence to a fixed advanced sleep/wake schedule resulted in significant circadian phase shifts in young adults with subclinical DSPD with or without morning blue light exposure. Light/dark exposures associated with fixed early sleep schedules are sufficient to advance circadian phase in young adults.

Effects on subjective and objective alertness and sleep in response to evening light exposure in older subjects

Evening bright light exposure is reported to ameliorate daytime sleepiness and age-related sleep complaints, and also delays the timing of circadian rhythms. We tested whether evening light exposure given to older adults with sleep–wake complaints would delay the timing of their circadian rhythms with respect to their sleep timing, thereby reducing evening sleepiness and improving subsequent sleep quality. We examined the impact of evening light exposure from two different light sources on subjective alertness, EEG activity during wakefulness, and sleep stages. Ten healthy older adults with sleep complaints (mean age = 63.3 years; 6F) participated in a 13-day study. After three baseline days, circadian phase was assessed. On the evening of days 5–8 the subjects were exposed for 2 h to either polychromatic blue-enriched white light or standard white fluorescent light, and on the following day circadian phase was re-assessed. Subjects were allowed to leave the laboratory during all but the two days when the circadian phase assessment took place. Evening assessments of subjective alertness, and wake and sleep EEG data were analyzed. Subjective alertness and wake EEG activity in the alpha range (9.75–11.25 Hz) were significantly higher during light exposures when compared to the pre-light exposure evening ( p < 0.05). The light exposures produced circadian phase shifts and significantly prolonged latency to rapid eye-movement (REM) sleep for both light groups ( p < 0.05). The increase in wake EEG alpha activity during the light exposures was negatively correlated with REM sleep duration ( p < 0.05). Evening light exposure could benefit older adults with early evening sleepiness, without negatively impacting the subsequent sleep episode.

Partial Sleep Deprivation Reduces Phase Advances to Light in Humans

Partial sleep deprivation is increasingly common in modern society. This study examined for the first time if partial sleep deprivation alters circadian phase shifts to bright light in humans. Thirteen young healthy subjects participated in a repeated-measures counterbalanced design with 2 conditions. Each condition had baseline sleep, a dim-light circadian phase assessment, a 3-day phase-advancing protocol with morning bright light, then another phase assessment. In one condition (no sleep deprivation), subjects had an 8-h sleep opportunity per night during the advancing protocol. In the other condition (partial sleep deprivation), subjects were kept awake for 4 h in near darkness (<0.25 lux), immediately followed by a 4-h sleep opportunity per night during the advancing protocol. The morning bright light stimulus was four 30-min pulses of bright light (~5000 lux), separated by 30-min intervals of room light. The light always began at the same circadian phase, 8 h after the baseline dim-light melatonin onset (DLMO). The average phase advance without sleep deprivation was 1.8 ± 0.6 (SD) h, which reduced to 1.4 ± 0.6 h with partial sleep deprivation (p < 0.05). Ten of the 13 subjects showed reductions in phase advances with partial sleep deprivation, ranging from 0.2 to 1.2 h. These results indicate that short-term partial sleep deprivation can moderately reduce circadian phase shifts to bright light in humans. This may have significant implications for the sleep-deprived general population and for the bright light treatment of circadian rhythm sleep disorders such as delayed sleep phase disorder.

Two Components of Nocturnal Locomotor Suppression by Light

In nocturnal rodents, millisecond light ("flash") stimuli can induce both a large circadian rhythm phase shift and an associated state change from highly active to quiescence followed by behavioral sleep. Suppression of locomotion ("negative masking") is an easily measured correlate of the state change. The present mouse studies used both flashes and longer light stimuli ("pulses") to distinguish initiation from maintenance effects of light on locomotor suppression and to determine whether the locomotor suppression exhibits temporal integration as is thought to be characteristic of phase shift responses to pulse, but not flash, stimuli. In experiment 1, locomotor suppression increased with irradiance (0.01-100 microW/cm( 2)), in accordance with previous reports. It also increased with stimulus duration (3-3000 sec), but interpretation of this result is complicated by the ability of light to both initiate and maintain locomotor suppression. In experiment 2, an irradiance response curve was determined using a stimulus series of 10 flashes, 2 msec each, with total flash energy varying from 0.0025 to 110.0 J/m(2). This included a test for temporal integration in which the effects of two equal energy series of flashes that differed in the number of flashes per series (10 vs 100), were compared. The 10 flash series more effectively elicited locomotor suppression than the 100 flash series, a result consistent with prior observations involving flash-induced phase shifts. In experiment 3, exposure of mice to an 11-h light stimulus yielded irradiance-dependent locomotor suppression that was maintained for the entire stimulus duration by a 100-microW/cm(2) stimulus. Light has the ability to initiate a time-limited (30-40 min) interval of locomotor suppression (initiation effect) that can be extended by additional light (maintenance effect). Temporal integration resembling that seen in phase-shifting responses to light does not exist for either phase shift or locomotor suppression responses to flashes or for locomotor suppression responses to light pulses. The authors present an alternative interpretation of data thought to demonstrate temporal integration in the regulation of phase shift responses to light pulses.

Sleep dependent learning in contour integration

Aim. A large body of evidence supports the involvement of sleep dependent mechanisms in perceptual learning (Karni et al, 1994; Stickgold et al, 2000). However, sleep dependence has only been studied extensively in the texture discrimination paradigm. We have indicated earlier (Kozma & Kovacs, 2002) that improvement in a contour integration task might also depend on sleep, but our design did not make a clear distinction between time vs. sleep dependent learning. Here we clarify the role of sleep in the contour integration task. Method. The contour integration task consisted of an egg-shaped, closed contour of Gabor patches on a background of randomly positioned and oriented Gabor patches. Subjects had to determine the direction where the egg-shape was pointing to at various levels of difficulty. 40 subjects practiced in the contour integration task over five 15 min sessions (240 trials/session) with 12 hours between sessions. We introduced a circadian phase shift between two groups of subjects. One group of subjects started the 1st session at 8 a.m. (Morning Group/MG), while the second group started at 8 p.m. on the same day (Evening Group/EG). Result. By the 4th session (2 nights of sleep for EG subjects, and only 1 night of sleep for MG subjects), EG performance was significantly better than MG performance. Conclusion. The ‘circadian shift’ design (see also Walker, 2002) allowed us to dissociate the impact of time vs. sleep on perceptual learning. We can now safely say that learning in the contour integration task is dependent on sleep.

Circadian entrainment to light–dark cycles involves extracellular nitric oxide communication within the suprachiasmatic nuclei

The ability to synchronize to light-dark (LD) cycles is an essential property of the circadian clock, located in mammals within the hypothalamic suprachiasmatic nuclei (SCN). Single light pulses activate nitric oxide (NO) intracellular signaling, leading to circadian phase-shifts required for synchronization. In addition, extracellular NO has a role in the SCN paracrine communication of photic phase advances. In this work, the extracellular nitrergic transmission was assessed in steady-state synchronization to LD cycles of locomotor rhythms in the golden hamster (Mesocricetus auratus). Extracellular NO levels were pharmacologically decreased in vivo with the specific scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO). Hamsters were subjected to LD cycles different from normal 24 h (LD 14 : 10) cycles (i.e. T-cycles), with a single 30-min light pulse presented either every 23 h (T23 cycles), or every 25 h (T25 cycles), thus allowing synchronization by advances or delays, respectively. Acute PTIO intracerebroventricular microinjections, delivered 30 min previous to the light pulse, inhibited synchronization by phase advances to T23 cycles, but did not alter phase delays under T25 cycles. In addition, NO scavenging inhibited light-induced expression of PERIOD1 protein at circadian time 18 (i.e. the time for light-induced phase advances). These findings demonstrate the role of extracellular NO communication within the SCN in the steady-state synchronization to LD cycles.

Millisecond Light Pulses Make Mice Stop Running, then Display Prolonged Sleep-Like Behavior in the Absence of Light

Masking, measured as a decrease in nocturnal rodent wheel running, is a visual system response to rod/cone and retinal ganglion cell photoreception. Here, the authors show that a few milliseconds of light are sufficient to initiate masking, which continues for many minutes without additional photic stimulation. C57J/B6 mice were tested using flash stimuli previously shown to elicit large circadian rhythm phase shifts. Ten flashes, 2 msec each and equally distributed over 5 min, activate locomotor suppression that endures for an additional 25 to 35 min in the dark and does not differ in magnitude or duration from that elicited by 5-min saturating light pulse. Locomotor activity by mice without access to running wheels is also suppressed by light flashes. The effects of various light flash patterns on mouse locomotor suppression are similar to those previously described for hamster phase shifts. Video analysis of active mice indicates that light flashes initiated at ZT13 rapidly induce an interval of behavioral quiescence that lasts about 10 min at which time the animals assume a typical sleep posture that is maintained for an additional 25 min. Thus, the period coincident with light-induced wheel running suppression appears to consist of two distinct behavioral states, one interval during which locomotor quiescence is initiated and maintained, followed by a second interval characterized by behavioral sleep. Given this sequence effected by light stimulation, we suggest that it be referred to as "photosomnolence," the term reflecting upon both the nature of the stimulus and the associated behavioral change.

Sleep Patterns in High School and University Students: A Longitudinal Study

We performed a longitudinal study to investigate whether changes in social zeitgebers and age alter sleep patterns in students during the transition from high school to university. Actimetry was performed on 24 high-school students (mean age ± SD: 18.4 ± 0.9 yrs; 12 females) for two weeks. Recordings were repeated in the same subjects 5 yrs later when they were university students. The sleep period duration and its center, the mid-sleep time, and total sleep time were estimated by actimetry. Actigraphic total sleep time was similar when in high school and at the university on school days (6.31 ± 0.47 vs. 6.45 ± 0.80 h, p = ns) and longer on leisure days by 1.10 ± 1.10 h (p < 0.0001 vs. school days) when in high school, but not at the university. Compared to the high school situation, the mid-sleep time was delayed when at the university on school days (03∶11 ± 0.6 vs. 03∶55 ± 0.7 h, p < 0.0001), but not on leisure days. Individual mid-sleep times on school and leisure days when in high school were significantly correlated with the corresponding values 5 yrs later when at the university (r = 0.58 and r = 0.55, p < 0.05, respectively). The large differences in total sleep time between school and leisure days when students attended high school and the delayed mid-sleep time on school days when students attended university are consistent with a circadian phase shift due to changes in class schedules, other zeitgebers, and lifestyle preferences. Age-related changes may also have occurred, although some individuality of the sleep pattern was maintained during the 5 yr study span. These findings have important implications for optimizing school and work schedules in students of different age and level of education. (Author correspondence: pneubloc@usz.uzh.ch)

SLEEP PATTERNS IN HIGH SCHOOL AND UNIVERSITY STUDENTS: A LONGITUDINAL STUDY

We performed a longitudinal study to investigate whether changes in social zeitgebers and age alter sleep patterns in students during the transition from high school to university. Actimetry was performed on 24 high-school students (mean age ± SD: 18.4 ± 0.9 yrs; 12 females) for two weeks. Recordings were repeated in the same subjects 5 yrs later when they were university students. The sleep period duration and its center, the mid-sleep time, and total sleep time were estimated by actimetry. Actigraphic total sleep time was similar when in high school and at the university on school days (6.31 ± 0.47 vs. 6.45 ± 0.80 h, p = ns) and longer on leisure days by 1.10 ± 1.10 h (p < 0.0001 vs. school days) when in high school, but not at the university. Compared to the high school situation, the mid-sleep time was delayed when at the university on school days (03∶11 ± 0.6 vs. 03∶55 ± 0.7 h, p < 0.0001), but not on leisure days. Individual mid-sleep times on school and leisure days when in high school were significantly correlated with the corresponding values 5 yrs later when at the university (r = 0.58 and r = 0.55, p < 0.05, respectively). The large differences in total sleep time between school and leisure days when students attended high school and the delayed mid-sleep time on school days when students attended university are consistent with a circadian phase shift due to changes in class schedules, other zeitgebers, and lifestyle preferences. Age-related changes may also have occurred, although some individuality of the sleep pattern was maintained during the 5 yr study span. These findings have important implications for optimizing school and work schedules in students of different age and level of education. (Author correspondence: pneubloc@usz.uzh.ch)

Chronic Ethanol Intake Alters Circadian Phase Shifting and Free-Running Period in Mice

Chronic alcohol intake is associated with widespread disruptions in sleep and circadian rhythms in both human alcoholics and in experimental animals. Recent studies have demonstrated that chronic and acute ethanol treatments alter fundamental properties of the circadian pacemaker--including free-running period and responsiveness to photic and nonphotic phase-shifting stimuli--in rats and hamsters. In the present work, the authors extend these observations to the C57BL/6J mouse, an inbred strain characterized by very high levels of voluntary ethanol intake and by reliable and stable free-running circadian activity rhythms. Mice were housed individually in running-wheel cages under conditions of either voluntary or forced ethanol intake, whereas controls were maintained on plain water. Forced ethanol intake significantly attenuated photic phase delays (but not phase advances) and shortened free-running period in constant darkness, but voluntary ethanol intake failed to affect either of these parameters. Thus, high levels of chronic ethanol intake, beyond those normally achieved under voluntary drinking conditions, are required to alter fundamental circadian pacemaker properties in C57BL/6J mice. These observations may be related to the relative ethanol insensitivity displayed by this strain in several other phenotypic domains, including ethanol-induced sedation, ataxia, and withdrawal. Additional experiments will investigate chronobiological sensitivity to ethanol in a range of inbred strains showing diverse ethanol-related phenotypes.

Phase delaying the human circadian clock with a single light pulse and moderate delay of the sleep/dark episode: no influence of iris color

BackgroundLight exposure in the late evening and nighttime and a delay of the sleep/dark episode can phase delay the circadian clock. This study assessed the size of the phase delay produced by a single light pulse combined with a moderate delay of the sleep/dark episode for one day. Because iris color or race has been reported to influence light-induced melatonin suppression, and we have recently reported racial differences in free-running circadian period and circadian phase shifting in response to light pulses, we also tested for differences in the magnitude of the phase delay in subjects with blue and brown irises.MethodsSubjects (blue-eyed n = 7; brown eyed n = 6) maintained a regular sleep schedule for 1 week before coming to the laboratory for a baseline phase assessment, during which saliva was collected every 30 minutes to determine the time of the dim light melatonin onset (DLMO). Immediately following the baseline phase assessment, which ended 2 hours after baseline bedtime, subjects received a 2-hour bright light pulse (~4,000 lux). An 8-hour sleep episode followed the light pulse (i.e. was delayed 4 hours from baseline). A final phase assessment was conducted the subsequent night to determine the phase shift of the DLMO from the baseline to final phase assessment.Phase delays of the DLMO were compared in subjects with blue and brown irises. Iris color was also quantified from photographs using the three dimensions of red-green-blue color axes, as well as a lightness scale. These variables were correlated with phase shift of the DLMO, with the hypothesis that subjects with lighter irises would have larger phase delays.ResultsThe average phase delay of the DLMO was -1.3 ± 0.6 h, with a maximum delay of ~2 hours, and was similar for subjects with blue and brown irises. There were no significant correlations between any of the iris color variables and the magnitude of the phase delay.ConclusionA single 2-hour bright light pulse combined with a moderate delay of the sleep/dark episode delayed the circadian clock an average of ~1.5 hours. There was no evidence that iris color influenced the magnitude of the phase shift. Future studies are needed to replicate our findings that iris color does not impact the magnitude of light-induced circadian phase shifts, and that the previously reported differences may be due to race.

Seasonal affective disorder and exercise treatment: a review

In recent years, there has been considerable improvement in the diagnostic criteria and epidemiology of seasonal affective disorder (SAD). This condition is a cyclically recurring mood and neuro-vegetative disorder that particularly affects populations residing in northern latitudes. It is characterised by regular depressive episodes in the autumn and winter, and non-depressive periods in the spring and summer. To date, the aetiology of SAD has not been identified. The leading hypotheses include seasonal variation of the photoperiod, circadian phase shift and neurotransmitter dysfunction. A number of recent studies suggest that physical exercise may provide an effective and easily accessible treatment for patients suffering from SAD. Despite growing research efforts, the palliative effects of exercise on depression and seasonal mood disorders are not clearly understood. This review assesses the clinical and physiological evidence which suggests that exercise intervention may facilitate effective treatment for SAD. Particular emphasis is given to circadian mechanisms that have been hypothesised to explain mood-enhancing affects of physical exercise.

Melatonin, sleep disturbance and cancer risk

The pineal hormone melatonin is involved in the circadian regulation and facilitation of sleep, the inhibition of cancer development and growth, and the enhancement of immune function. Individuals, such as night shift workers, who are exposed to light at night on a regular basis experience biological rhythm (i.e., circadian) disruption including circadian phase shifts, nocturnal melatonin suppression, and sleep disturbances. Additionally, these individuals are not only immune suppressed, but they are also at an increased risk of developing a number of different types of cancer. There is a reciprocal interaction and regulation between sleep and the immune system quite independent of melatonin. Sleep disturbances can lead to immune suppression and a shift to the predominance in cancer-stimulatory cytokines. Some studies suggest that a shortened duration of nocturnal sleep is associated with a higher risk of breast cancer development. The relative individual contributions of sleep disturbance, circadian disruption due to light at night exposure, and related impairments of melatonin production and immune function to the initiation and promotion of cancer in high-risk individuals such as night shift workers are unknown. The mutual reinforcement of interacting circadian rhythms of melatonin production, the sleep/wake cycle and immune function may indicate a new role for undisturbed, high quality sleep, and perhaps even more importantly, uninterrupted darkness, as a previously unappreciated endogenous mechanism of cancer prevention.

Interaction of chronic sleep restriction and circadian system in humans

Nocturnal sleep restriction and compensation with daytime naps is common in today's society. In a between-participants design, we examined the effects of chronic (10 nights) sleep restriction on 24 h plasma melatonin profiles in humans. Following a baseline period with 8.2 h time in bed (TIB) for sleep, participants were randomized to a control (8.2 h TIB) or sleep-restriction condition (4.2 h TIB), with and without diurnal naps. Sleep restriction was achieved via delaying bedtime and advancing wake time by 2 h each relative to the baseline sleep period. Participants were maintained in a controlled, time isolated laboratory environment throughout the protocol, with light levels below 40 lx at all times. Twenty-four hour plasma melatonin profiles were assessed at baseline and at the end of the sleep-restriction period, with subjects maintained in a constant posture protocol. Compared with the baseline assessment and the 8.2 h TIB control group, a significant phase delay in melatonin onset (1.2 +/- 0.9 h) occurred in all sleep-restriction (4.2 h TIB) groups (P < 0.05). There was no evidence of a phase advance or shortening of the period of melatonin secretion associated with the advanced waking time. These results suggest that nocturnal light and dark exposure may be more potent in effecting circadian phase shifts than exposure to morning light, at least in conditions of controlled, dim lighting in the laboratory.

Circadian Phase-Shifting Effects of Repeated Ramelteon Administration in Healthy Adults

To assess the ability of repeated daily oral ramelteon to facilitate re-entrainment of human circadian rhythms after an imposed phase advance of the sleep-wake cycle. A total of 75 healthy adult volunteers aged 18-45 years remained in a sleep laboratory for 6 days and 5 nights; a 5-h phase advance in their sleep-wake cycle was imposed under dim-light conditions. Oral ramelteon (1,2, 4, or 8 mg once daily for 4 days) or placebo was administered 30 min before bedtime. The primary endpoint was the phase of the circadian rhythm as assessed by the time at which salivary melatonin concentrations declined below 3 pg/mL after morning awakening (dim-light melatonin offset [DLMoff]). After 4 days of once-daily treatment, participants receiving 1, 2, or 4 mg ramelteon exhibited statistically significant phase shifts in DLMoff of -88.0 (16.6), -80.5 (14.8), and -90.5 (15.2) minutes respectively, versus -7.1 (18.6) minutes for placebo (least-squares mean(SEM), p = 0.002, p = 0.003, p = 0.001, respectively). Change in DLMoff for the 8 mg dose of ramelteon, -27.9 (16.4) minutes, was not significantly different than that for placebo (p = 0.392). Ramelteon (1, 2, or 4 mg per day) administered before bedtime significantly advanced the phase of the circadian rhythm after a 5-h phase advance in the sleep-wake cycle. These findings suggest that ramelteon has potential as a specific therapy for circadian rhythm sleep disorders.

Immunosuppressant calcineurin inhibitors phase shift circadian rhythms and inhibit circadian responses to light

PP2B is a Ca 2+/calmodulin-dependent protein phosphatase that is ubiquitously expressed in mammals. Among other actions, it is an effector mechanism in NMDA-mediated glutamate neurotransmission as well as a regulator of GSK3β and MAPK signaling cascades. Because all of these mechanisms have demonstrable roles in the control of circadian rhythyms, we hypothesized that PP2B would be a key regulator of rhythm generation and entrainment, and that through inhibition of its phosphatase activity, the circadian system would be affected by immunosuppressant drug therapy. We report here that immunosuppressant drugs (cyclosporin A, FK506) (1) block the circadian responses to light that underlie photic entrainment; (2) produce circadian phase shifts with a characteristic nonphotic profile; and (3) disrupt circadian rhythm expression when applied chronically. These results indicate a role for PP2B in circadian rhythm generation and entrainment. In addition, because rhythm disturbance has been implicated in impairment of both physical and mental health, we suggest that the use of immunosuppressants would be safer and more efficacious if their impacts on circadian rhythmicity were taken into account.