[The circadian system in man: From the internal clock to melatonin secretion]. / À propos du système circadien chez l'homme : de l'horloge interne à la sécrétion de mélatonine.

The internal or biological clock which is located in the suprachiasmatic nuclei of the anterior hypothalamus is controlled by clock genes and environmental factors which are able to synchronize the clock to 24h. Rhythm desynchronization (shiftwork and nightwork, transmeridian flights, aging, some psychiatric diseases, blindness, intake of some drugs…) occurs when the internal clock does no longer work in harmony with the astronomical time i.e. our watch. The circadian system consists of three major elements, which are the clock, the retinohypothalamic tract and melatonin which is secreted by the pineal gland and considered as the arrow of the clock. Both light and melatonin present a phase response curve useful for the treatment of sleep circadian disorders.

Chronobiological aspects of food intake and metabolism and their relevance on energy balance and weight regulation.

Overweight and obesity are the result of a chronic positive energy balance, and therefore the only effective therapies are a diet which, on the long term, provides lower calories than the daily expended energy and exercise. Because nearly every physiological and biochemical function of the body shows circadian variations it can be suggested that also different chronobiological aspects of food intake, like time of day, meal frequency and regularity, and also circadian desynchronizations like in shift work may affect energy metabolism and weight regulation. The aim of this review is therefore to summarize and discuss studies that have addressed these issues in the past and to also provide an overview about circadian variations of selected aspects of metabolism, gut physiology and also factors that may influence overall energy regulation. The results show that a chronic desynchronization of the circadian system like in shift work and also sleep deprivation can favour the development of obesity. Also, regarding energy balance, a higher meal frequency and regular eating pattern seem to be more advantageous than taking the meals irregularly and seldom. Additional studies are required to conclude whether time of day-dependent food intake significantly influences weight regulation in humans.

Light pulse induces ALA-S gene expression in the rat Harderian gland.

The rodent Harderian glands (HGs) are large paired orbital organs with highest porphyrinogenic rates. We have previously shown that continuous light exposure abolished the day/night variations of the delta-aminolevulinate synthase (ALA-S; the rate-limiting enzyme for porphyrin biosynthesis) gene expression observed under standard light: dark cycles (LD 12:12) in the rat HGs. This study was designed to examine whether the ALA-S changes were actually associated directly with light. The response of ferrochelatase (enzyme that converts protoporphyrin IX into heme) to light was also examined. Male Wistar rats were acclimatized to light: dark cycles regimen of 12:12 for 2 weeks. At the end of the 2 weeks, a 1 h-light pulse was applied in the middle of the dark phase. Animals were sacrificed immediately after the end of the light pulse. HGs were collected and stored at -80 degrees C until processed for quantitative RT-PCR. A 1 h-light pulse applied during mid-dark caused a significant increase of ALA-S gene expression (3-fold higher than in controls), whereas it was without effect on ferrochelatase gene expression. Our results suggest that light per se may regulate ALA-S gene expression in the rat HGs, and reveal that the ALA-S gene expression, and so heme biosynthesis, is under a photodynamic control.

Desynchronization of daily rest-activity rhythm in the days following light propofol anesthesia for colonoscopy.

Anesthesia and surgery are associated with fatigue and sleep disorders, suggestive of disturbance of the circadian rest-activity rhythm. Previous studies on circadian rhythm disturbance were focused on patients undergoing general anesthesia associated with surgery. This does not permit one to draw valid conclusions about the effects of general anesthesia per se on circadian rhythms. Our study was set up to determine the impact of a hypnotic dose of propofol on the circadian rest-activity rhythm in humans under real-life conditions. Seventeen healthy subjects scheduled to receive light propofol anesthesia for ambulatory colonoscopy were investigated. Their rest-activity rhythms were assessed using actigraphic monitoring. Diurnal rest was increased, whereas nocturnal sleep was unchanged in the days following anesthesia. Nonparametric analyses showed a decrease in the strength of coupling of the rhythm to stable environmental zeitgebers and increase of fragmentation of the rhythm after anesthesia. Light general anesthesia itself impairs synchronization of the circadian rest-activity rhythm to local time in patients by acting directly on the circadian clock.

[Dysfunctions of biological clocks and their treatments]. / Dysfonctionnements de l'horloge biologique et leurs traitements.

Biological rhythms are periodic phenomena entrained to environmental changes by exogenous factors called synchronizers or entraining agents namely the light-dark cycle, the rest-activity cycle and the seasons, among others. In humans the major synchronizers are the light-dark and rest activity cycles. The endogenous component of a biological rhythm is dependent upon a number of clock genes. The main biological clock (oscillator or pacemaker) is the suprachiasmatic nucleus (SCN) of the anterior hypothalamus. The photoperiod (light-dark cycle) perceived by the retina acts on the SCN genes. Peripheral clocks have also been described in a number of tissues e.g. retina, adrenals. In a number of occurrences the synchronizers are badly perceived (transmeridian flights, shiftwork, nightwork...) or are not at all perceived (blindness). This situation is named rhythm desynchronization, it is external when the desynchronization is strictly related to the environment or internal when it is related to a dysfunction of the clock like in e.g. aging, Alzheimer disease, seasonal affective disorders (SAD) or hormone-dependent cancers which results in fatigue, sleep and mood disorders... A number of drugs called resynchronizing agents or chronobiotics which act on the biological clock are able to resynchronize the clock and to improve the patients' condition. Bright light is used in the treatment of SAD, melatonin, the pineal hormone, is also of interest when administered at precise timings in the 24hours scale. Other drugs like B12 vitamin or psychotropic drugs have also been proposed as chronobiotics.

[Sleep disorders and hypnotic agents: medical, social and economical impact]. / Troubles du sommeil et hypnotiques: impacts médicaux et socio-économiques.

Insomnia is a subjective complaint relating to approximately 30% of the adult population in France, described by the patient as a difficulty of initiating and/or maintaining sleep. Its prevalence increases with age and sex: women are more affected than men (24% vs 14%). Insomnia is either occasional (20%), or chronic (10%). Chronic insomnia has an important impact on patients' everyday life e.g. fatigue, perturbed diurnal waking state, impaired quality-of-life... which results in lower work productivity and drowsiness as well as relational difficulties, absenteeism. About 80% of patients consult their general practitioner first. The aim of a hypnotic agent is to obtain sleep as physiological as possible. Benzodiazepines and benzodiazepines-like agents (zopiclone, zolpidem, zaleplon) are the most widely used hypnotics. However, their indications must be limited to occasional insomnia with a limited duration: less than four weeks. There is no advantage with using a combination of hypnotic agents, a practice which should be prohibited. Adverse effects can be serious, e.g. diurnal somnolence associated with risks of road accidents and, in the elderly, the risk of falls. After chronic use, hypnotics can be addictive, as their effects wear off in three to four weeks. After withdrawal, insomnia rebound is frequent. Use of hypnotics in association with alcohol is a well-known drug-addiction behavior. According to the French health insurance fund, 9% of the general population use hypnotics and about half of them regularly. Insurance refunds for hypnotics and sedatives reach more than 110 million euros annually. The efficiency of hypnotics wears off, quickly for benzodiazepines (three - four weeks), or less quickly for zopiclone and zolpidem (a few months). Insomnia is a major public health issue, each year 10% of the incident cases of insomnia treated by hypnotics joint the group of subjects with chronic insomnia. This failure to treat insomnia properly can be explained, at least in part, by several insufficiencies: physicians and pharmacists training, medical profession awareness, research, public information on the rules of good sleep (public health campaigns, booklets, role of physicians and the pharmacists).

Is melatonin the hormonal missing link between magnetic field effects and human diseases?

The disruption of melatonin secretion has been largely studied since it could provide the missing link between the exposure to 50/60-Hz electric and magnetic fields (EMF) and the occurrence of possible health effects as the "melatonin hypothesis". We analysed the current experimental data from animal (rodents) where contradictory results have been observed, and from human studies conducted with volunteers or with workers in various conditions of exposure, biological endpoints and metrics. In humans, even in long lasting exposures, the overall results of these studies do not support the "melatonin hypothesis". It is unlikely that malignancies or mood disorders reported by people exposed to 50/60-Hz EMF could be related to the disruption of the melatonin levels.

Impairment in clock-time estimation following right hemisphere ischemic damage.

In order to assess clock-time estimation (CTE), we asked "what time is it in your opinion?" to 48 recent stroke in-patients, 21 with right (RH), 27 with left hemispheric (LH) lesions, and to 20 control in-patients without brain lesions (C). Errors were measured in terms of the number of minutes by which the estimated clock-time was later (advance errors) or earlier (delay errors) than the real clock-time. CTE was considered pathological when exceeding the mean advance or delay errors observed in control patients plus 2.5 standard deviations. The estimation of the duration of a short psychological interview was also assessed. CTE, and not duration estimation, was disturbed in patients. RH patients made significantly more pathological advance errors than LH patients (43% vs. 12%). This study points out the RH dominance for CTE in stroke patients.

Prolactin rhythms-oscillators' response to photoperiodic cues is age and circadian time dependent.

In vivo prolactin release patterns exhibit a compound rhythm with circadian (24 h), semicircadian (12 h) and ultradian (6-8 h) periods. Changes in these rhythmic patterns were observed at different photoperiodic conditions, and in elderly. Since in vitro prolactin release was related to the photoperiodic history of the animal, we studied the effect of varying photoperioda upon the in vitro rhythmic output of prolactin release from young and old male rat pituitaries, isolated at different circadian times from animals housed at LD 12:12, 18:6 for 10 days or 6 weeks. The results indicate that, both, mean levels and rhythmic prolactin release in vitro are determined by the age of the animal, the circadian time of pituitary isolation, the photoperiodic conditions in which the animal was housed, and the duration of housing in the long day conditions. The change of the rhythmic output pattern is gradual, reflecting a process by which the oscillators respond to the external cues to fit prolactin release pattern to the environmental conditions. Each of the oscillators (e.g. circadian, semicircadian, ultradian) shows different sensitivity to the changing photoperiodic signal and is regulated at the level of phase and amplitude but not the period. In old rats the response of the oscillators to the change in photoperioda is attenuated and not sufficient to induce a change in the output of prolactin release suggesting a loss in adaptation ability.

[Evaluation of the effects of electric and magnetic fields in humans]. / Evaluation des effets des champs électromagnétiques sur la santé chez l'homme.

During the twentieth century, environmental exposure to electromagnetic fields generated by human activity has increased regularly, at the same time as the quest for electrical energy. Therefore we are all exposed to a complex set of electric and magnetic fields of weak intensity. The levels of exposure of the general population range from 5 to 50 V/m for electric fields and from 0.01 to 0.2 micro T for magnetic fields. Fields in cause are essentially those associated to the use of the electric current (extremely low frequency, ELF: 50 Hz in France, 60 Hz in the United States) and those related to the use of cell phones (radio frequency: 900 and 1,800 MHz). The question of the possible risk on health by exposure to electric and/or magnetic fields became a concern to scientists and is now an important public debate. A number of expertises, led in particular by the WHO, leaning on the careful inspection of scientific publications from numerous countries, conclude that current data do not allow to assert the existence of sanitary effects; however our knowledge of the biologic effects of electromagnetic fields still contains certain gaps which should be filled. Indeed, the numerous epidemiological studies relative to the occurrence of cancer by exposure to electromagnetic fields are conflicting. In every case the increase of the risk, when described, is always weak. The measure of the Relative Risk (RR) which establishes the relation is approximately 2-3. At present, some data concerning the risk of childhood leukemia in the event of exposure of ELF generated in the home indicate that this risk can exist when children are chronically exposed to more than 0.4 micro T (the relative risk is in the order of 2). In the field of radio frequencies, the increasing use of cell phones (38 million users in France) and their antennae - basis are another subject of concern for the effects on health they are susceptible to produce. Large-scale studies, implying numerous countries, carried on at present within the framework of the WHO should bring elements of answer to the unresolved questions.

Evaluation of the nocturnal levels of urinary biogenic amines in men exposed overnight to 50-Hz magnetic field.

The aim of this study was to determine whether exposure to magnetic fields might affect human health and to look for possible effects of acute exposure (9 hours) to 50-Hz magnetic fields (10 microT) on the urinary concentration of biogenic amines. Thirty-two young men (20-30 years old) were divided into two groups (sham-exposed and exposed group) of 12 to 16 subjects each. All subjects participated in two 24-hour experiments to evaluate the effects of both continuous and intermittent exposure to magnetic fields. The subjects were exposed to the magnetic field from 2300 to 0800, while lying down. Total urine (from 2300 to 0800) was collected at 0800. The results (expressed as a ratio of biogenic amine excretion to creatinine excretion (nmol/mmol)) did not differ significantly between sham-exposed and exposed men for any of the parameters measured: adrenaline, noradrenaline, dopamine, dihydroxyphenylalanine, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid. These results suggest that nocturnal exposure to either continuous or intermittent 50-Hz magnetic fields of 10 microT does not affect, at least under our experimental conditions, the nocturnal excretion of biogenic amines in healthy young men.

Magnetic field (50 Hz) increases N-acetyltransferase, hydroxy-indole-O-methyltransferase activity and melatonin release through an indirect pathway.

PURPOSE: To examine whether magnetic fields (MF) affect N-acetyltransferase (NAT) and hydroxy-indole-O-methyltransferase (HIOMT) activity directly or exert their effect through a cellular pathway that indirectly regulates the activity of these enzymes and melatonin release. MATERIALS AND METHODS: The pineal glands from Wistar rats were isolated at 10:00 h and exposed to MF (50 Hz, 1 mT) for 4 h in vitro, with or without 1 micro M norepinephrine. An additional group of pineals was exposed to MF 30 min before norepinephrine addition. The direct effect of MF on the activity of the enzymes was studied in sonicated glands exposed to MF. NAT activity, HIOMT activity and melatonin release were determined. RESULTS: In pineal glands isolated in the morning, 4-h in vitro exposure did not affect the basal release of melatonin from the pineal gland as well as the basal NAT and HIOMT activities. Pineal gland exposure to MF 30 min before norepinephrine addition significantly (p<0.05) increased NAT activity, HIOMT activity and melatonin release (p<0.05). These effects were not observed in pineals co-treated with MF and norepinephrine or in sonicated glands exposed to MF. CONCLUSIONS: The results suggest that in pineals isolated in the morning, 4-h MF exposure changes melatonin release by affecting the signal transduction pathway leading from the norepinephrine receptor to NAT and HIOMT and not via a direct effect at the enzyme levels.

Effect of a synthetic pineal tetrapeptide (Ala-Glu-Asp-GLy) on melatonin secretion by the pineal gland of young and old rats.

The pineal gland contains many peptides known to be implicated in melatonin production. We examined the effects of a synthetic pineal tetrapeptide Ala-Glu-Asp-Gly on melatonin secretion by the pineal gland. The tetrapeptide effects on pineal gland melatonin secretion were studied in young (9 weeks) and old (27 months) male Wistar rats using a perifusion device. Pineal tetrapeptide at the concentrations used (10(-4) to 10(-6) M) had no significant effect upon melatonin secretion whatever the age of the animals, young or old. We also looked at the effect of the tetrapeptide on pineal melatonin stimulated by a beta-adrenergic agonist, isoproterenol. We found that isoproterenol-induced melatonin increase was not modified by the tetrapeptide. Our results suggest that the pineal tetrapeptide Ala-Glu-Asp-Gly, does not seem to play a role, at least in vitro, in the control of melatonin secretion by the rat pineal gland.

Effect of age and photoperiodic conditions on metabolism and oxidative stress related markers at different circadian stages in rat liver and kidney.

It has been shown that some cytochrome P450-dependent enzyme activities could present daily fluctuations, particularly CYP3A isoenzymes which are enhanced during the dark period. The aim of this study was to investigate whether age and photoperiodic conditions at different circadian stages could influence these fluctuations. Young mature (10 weeks) and old (22 months) Wistar rats were initially exposed to light-dark cycles 12:12 during 4 weeks, and secondly 18:6 for either one week or six weeks. Erythromycin N-demethylase (CYP3A-dependent), 7-ethoxycoumarin O-deethylase (CYP1A-dependent) and aniline 4-hydroxylase (CYP2E-dependent) activities were determined in liver and kidney microsomes at different hours after darkness onset (HADO). In addition, liver and kidney GSH, GSHPx, ATP, TBARS were determined. During the LD 12:12 cycle, while no significant modification was observed in CYP1A- and 2E-dependent enzyme activities as functions of HADO, erythromycin N-demethylase activity (CYP3A-dependent) showed a significant increase during the second third of the dark period in both young and old rats. After switching to a LD 18:6 cycle, this variation was still observed during second third of the dark period, to a lesser but still significant degree, with no difference between one week and six weeks exposure to the new photoperiod. It can be noted that the old rats showed a significantly lower level of erythromycin N-demethylase activity than the young rats, in parallel to a decrease in GSH, GSHPx and ATP, and an increase in TBARS. These results confirm the lower resistance of old animals to oxidative stress. The observed variations in metabolism parameters underline the need for study designs in pharmaco-toxicology taking into account the possible risks induced by circadian changes, especially in aged subjects.

Study of circadian melatonin secretion pattern at different stages of Parkinson's disease.

To explore changes in melatonin secretion patterns and biologic rhythms in Parkinson's disease patients with or without levodopa-related motor complications (LDRMCs), the authors investigated, in an observational study, circadian rhythms of central temperature, motor activity, plasma cortisol, and melatonin in three groups: de novo untreated patients (group I), patients treated with levodopa + dopamine agonist and without LDRMCs (group II), and patients treated with levodopa + dopamine agonist and with LDRMCs (group III). There were no differences among the three groups for the rhythm of temperature, motor activity, or plasma cortisol. There was a significant (p < 0.05) phase advance in plasma melatonin secretion in patients receiving a dopaminergic treatment compared with untreated patients. The daytime area under the curve (AUC) was increased significantly in group III, and the nighttime AUC-to-daytime AUC ratio of melatonin secretion decreased significantly in group III, suggesting that the nychthemeral pattern of melatonin secretion was changed in patients with LDRMCs. Comparison of the three groups suggests a slight but insignificant phase advance and amplitude decrease of circadian melatonin secretion related to both evolution and treatment of Parkinson's disease. Despite the lack of a global desynchronization in other circadian biologic rhythms, the circadian secretion pattern of melatonin is modified in patients with LDRMCs.

Effect of a short photoperiod on circadian rhythms of body temperature and motor activity in old rats.

Circadian rhythms of body temperature and motor activity were documented in young and old rats (four 8-week-old and five 22-month-old male Wistars, implanted with telemetric probes and housed in a chronobiological facility) under two different photoperiod conditions. The animals were maintained in a light:dark (LD) cycle of 12 h each (LD 12:12) for 4 weeks and then exposed to a LD 6:18 cycle for 7 weeks to assess the effect of age on the desynchronization of the temporal structure of the rhythms. In old rats under LD 12:12, the power of the 24-h component and the circadian amplitude of body temperature and motor activity were markedly lower than in the young and both rhythms were phase-advanced. After the shift to LD 6:18, the circadian rhythmicity was maintained for both variables and the same phase delay (+5+/-1 h) was observed in both age groups, as was a gradual expansion of the patterns of both functions with the longer night. The photoperiod reduction (6 weeks under LD 6:18) did not modify the power of the 24-h component of body temperature and motor activity in old rats. In young rats, however, the power and amplitude of the 24-h component of motor activity rhythm fell to the levels of those in old rats, while the power of the 24-h component of body temperature rhythm and the amplitude did not change. Our data show that the circadian rhythm of motor activity, but not of body temperature, responds age dependently to a photoperiod reduction.