Rhythmic profiles of cell cycle and circadian clock gene transcripts in mice: a possible association between two periodic systems.

The circadian system shapes the rhythms of most biological functions. The regulation of the cell cycle by a circadian clock was suggested to operate via stages S, G2 and G2/M. This study investigated a possible time link at stages G1 and G1/S as well. The daily expression profiles of cell cycle markers (Ccnd1, Ccne1 and Pcna) and circadian clock genes (Per2 and Clock) were monitored in liver and esophagus (low and high proliferation index, respectively) of BALB/c mice. Locomotor activity displayed a 24 h rhythm, establishing the circadian organization of the suprachiasmatic nucleus. In the liver, the mRNA level of Per2 and Clock fitted the circadian rhythm with a 7.5 h shift. This temporal pattern suggests that the liver harbors a functional circadian clock. The rhythm of the analyzed cell cycle genes, however, was of low significance fitness and showed an opposite peak time between Pcna and Clock. These results indicate a weak regulatory role of the circadian clock. In the esophagus, the rhythms of Clock and Per2 mRNA had a similar peak time and non-circadian periods. These results suggest either that the esophagus does not harbor a functional circadian apparatus or that the phenotypes stem from differences in phase and amplitude of the rhythms of its various cell types. The similarity in the rhythm parameters of Clock, Ccne1 and Pcna transcripts questions the control of the circadian clock on the cell cycle along the G1 and G1/S stages. Yet the G1/S transition may play a role in modulating the local clock of proliferating tissues.

Cisplatin effects on rhythmic functions of mice: strain and tissue dependence.

The competence to preserve the optimal timing relationships between rhythmic variables enables adaptation of mammals to alternate environmental conditions. The capability to re-entrain depends on genetic factors and the nature of imposed time cues. In the present study, the authors examined in rodent models, following a cancer chronochemotherapy, cisplatin (CP), the rhythm patterns of locomotor activity and of a few biochemical variables (alkaline phosphatase and creatinine phosphokinase in kidney tissue and plasma, kidney urea nitrogen, and white blood cell count). Males of two inbred mice strains, BALB/c and c57Bl/6J, received 10 consecutive daily intraperitoneal (i.p.) injections of either saline or CP at zeitgeber time 22 (ZT22). CP administration altered the rhythms of each examined function in both strains. The type and extent of the changes varied among variables, tissues/plasma, and mouse strain. Yet, the effect of CP was not detected on all parameters, but only in ∼60% of them. In addition, in the majority of the studied parameters, BALB/c and c57Bl/6J mice differed in their response to CP. The temporal parameters of period and peak time were more affected by CP than were the level ones of mesor (time series mean) and amplitude of variation. This observation may indicate the involvement of independent pathways of action upon each of the rhythm parameter sets. As a result, the rhythm phenotype of each function was modified and novel timing relationships were shaped. The results show that the circadian systems of BALB/c and c57Bl/6J mice failed to re-entrain after cessation of CP injections (tested on the first day following the 10 d course of CP administration), pointing to a direct effect of the medication on the tissues. The findings imply that optimal chemotherapeutic protocols should be tailored individually, according to the current temporal order rather than administered at a fixed predetermined circadian time. Further studies are necessary to determine which variables and rhythmic parameters could be useful to determine the optimal timing of chronochemotherapy.

Internal desynchronization of circadian rhythms and tolerance to shift work.

Intolerance to shift work may result from individual susceptibility to an internal desynchronization. Some shift workers (SW) who show desynchronization of their circadian rhythms (e.g., sleep-wake, body temperature, and grip strength of both hands) exhibit symptoms of SW intolerance, such as sleep alteration, persistent fatigue, sleep medication dependence, and mood disturbances, including depression. Existing time series data previously collected from 48 male Caucasian French SW were reanalyzed specifically to test the hypothesis that internal synchronization of circadian rhythms is associated with SW intolerance and symptoms. The entry of the subjects into the study was randomized. Three groups were formed thereafter: SW with good tolerance (n=14); SW with poor tolerance, as evident by medical complaints for at least one year (n=19); and former SW (n=15) with very poor tolerance and who had been discharged from night work for at 1.5 yr span but who were symptom-free at the time of the study. Individual and longitudinal time series of selected variables (self-recorded sleep-wake data using a sleep log, self-measured grip strength of both hands using a Colin Gentile dynamometer, and oral temperature using a clinical thermometer) were gathered for at least 15 days, including during one or two night shifts. Measurements were performed 4-5 times/24 h. Power spectra used to quantify the prominent period (tau) and t-test, chi square, and correlation coefficient were used as statistical tools. The mean (+/-SEM) age of SW with good tolerance was greater than that of SW with poor tolerance (44.9+/-2.1 yrs vs. 40.1+/-2.6 yrs, p<.001) and of former SW discharged from night work (very poor tolerance; 33.4+/-1.7, p<.001). The shift-work duration (yrs) was longer in SW with good than poor tolerance (19.9+/-2.2 yrs vs. 15.7+/-2.2; p<0.002) and former SW (10.7+/-1.2; p<.0001). The correlation between subject age and shift-work duration was stronger in tolerant SW (r=0.97, p<.0001) than in non-tolerant SW (r=0.80, p<0.001) and greater than that of former SW (r=0.72, p<.01). The mean sleep-wake rhythm tau was 24 h for all 48 subjects. The number of desynchronized circadian rhythms (tau differing from 24 h) was greater in non-tolerant than in tolerant SW (chi square=38.9, p<.0001). In Former SW (i.e., 15 individuals assessed in follow-up studies done 1.5 to 20 yrs after return to day work), both symptoms of intolerance and internal desynchronization were reduced or absent. The results suggest that non-tolerant SW are particularly sensitive to the internal desynchronization of their circadian time organization.

Euchronism, allochronism, and dyschronism: is internal desynchronization of human circadian rhythms a sign of illness?

The authors define a subject as euchronic when the circadian parameters--tau (tau=period), Ø (acrophse or peak time), A (amplitude), and M (MESOR=24 h rhythm-adjusted mean)--of a set of circadian variables are within the confidence limits of appropriate reference values of healthy subjects (HS). We define internal desynchronization as a state in which the circadian tau of a set of rhythms differs from 24 h and when the tau of a given variable differs from that of other variables. Such a state was first observed in singly isolated HS without access to time cues and clues. Herein, data and analyses are presented demonstrating that internal desynchronization appears to be a rather common phenomenon in HS dwelling in their natural environment (i.e., in the presence of usual zeitgebers). This has been documented by longitudinal studies (n approximately=15 days) of the circadian rhythm in sleep-wakefulness, body temperature, right- and left-hand-grip strength, and reaction time involving a total of 246 HS and 134 shift workers (SW), with 45.5% showing good and 54.5% poor SW tolerance. The presence of internal desynchronization observed in SW was associated SW intolerance, with symptoms being sleep alteration/disturbances, sleeping-pill dependence, persisting fatigue (asthenia), mood alteration, and digestive complaints. Internal desynchronization was also documented in groups of HS and tolerant SW, though it was almost the rule among the intolerant SW. The authors introduce two new terms: allochronism to describe the time organization of those SW who evidence internal desynchronization without detectable clinical symptoms, and dyschronism to describe the time organization of those SW who exhibit internal desynchrobization plus the symptoms of SW intolerance or medical illness. The condition of allochronism is not restricted only to SW tolerance, as it was detected in 112 HS without medical complains when exposed to various experimental conditions, including medications and placebos, sojourn in the high Arctic summer, intensive sport training, and task-loaded cognitive performance testing. Dyschronism in SW who are sleep-deprived is associated with persisting fatigue. An unpublished Gallup survey found that 47% of 2478 respondents experienced a state of asthenia during the previous 12 months, with symptoms mimicking those of SW intolerance. In one-third of the cases, the origin of the asthenia was undetermined. Taking into account the high incidence of internal desynchronization found in past investigations and the clinical observation that sleep deprivation is a consequence of many acute and chronic medical conditions (nocturnal pain, nocturnal asthma, etc.), it is suggested that dyschronism may be responsible for the asthenia of unknown origin, at least for some persons. The interindividual (including sex-related) variability in the propensity to exhibit an altered temporal organization, whether it be transient or persistent (i.e., reversible or non-reversible) suggests the involvement of genetic factors. The Dian-Circadian genetic model previously proposed by the authors seems pertinent to conceptualize and explain the various levels and output of internal desynchronization.

Possible linkage between the ability to change the period (tau) of the prolactin and cortisol rhythms in women and breast cancer risk.

The 24 h profiles of plasma hormone concentrations are rhythmic. The circadian period (tau) changes in development, with seasons, and in women with different stages of the menstrual cycle. It is known that the rhythms of prolactin and cortisol are sensitive to environmental time cues, such as changes in day length and phase; however, the importance of these changes is not yet understood. This study investigates whether there is a relation between the ability of a subject to respond to external cues that are associated with seasonal changes causing alteration of the rhythm's periods in cortisol and prolactin and the epidemiologically determined susceptibility to breast cancer. It is shown that the rhythmic output pattern of prolactin and cortisol in vivo is generated by more than one oscillator and structured by more than one rhythmic component. Each cohort of American women, classified on an epidemiologic basis as high risk (HR) or low risk (LR) to develop breast cancer, expresses different rhythmic output patterns of both variables, suggesting that the genetic background as defined by the risk state is related to differences in the circadian time structure, including the ability of the subject to change the rhythm's tau. The LR cohort exhibited a statistically significant change between seasons in the rhythm's tau of both the prolactin and cortisol patterns. In contrast, the HR cohort showed no change in the rhythm's tau between seasons for prolactin and cortisol patterns. These results show that in human beings, the presence of a circannual rhythm in the circadian time structure or the ability to adapt the circadian rhythmic pattern of these variables to external cues, such as seasons, is related to the partly genetically determined risk state to develop breast cancer and may be of importance for human health.

Concepts in human biological rhythms.

Biological rhythms and their temporal organization are adaptive phenomena to periodic changes in environmental factors linked to the earth's rotation on its axis and around the sun. Experimental data from the plant and animal kingdoms have led to many models and concepts related to biological clocks that help describe and understand the mechanisms of these changes. Many of the prevailing concepts apply to all organisms, but most of the experimental data are insufficient to explain the dynamics of human biological clocks. This review presents phenomena thai are mainly characteristic ofand unique to - human chronobiology, and which cannot be fully explained by concepts and models drawn from laboratory experiments. We deal with the functional advantages of the human temporal organization and the problem of desynchronization, with special reference to the period (τ) of the circadian rhythm and its interindividual and intraindividual variability. We describe the differences between right- and left-hand rhythms suggesting the existence of different biological clocks in the right and left cortices, Desynchronization of rhythms is rather frequent (one example is night shift workers). In some individuals, desynchronization causes no clinical symptoms and we propose the concept of "allochronism" to designate a variant of the human temporal organization with no pathological implications. We restrict the term "dyschronism" to changes or alterations in temporal organization associated with a set of symptoms similar to those observed in subjects intolerant to shift work, eg, persisting fatigue and mood and sleep alterations. Many diseases involve chronic deprivation of sleep at night and constitute conditions mimicking thai of night shift workers who are intolerant to desynchronization. We also present a genetic model (the dian-circadian model) to explain interindividual differences in the period of biological rhythms in certain conditions.

Chemotoxicity and survival of tumor-bearing mice under exposure to randomized photoperiodic regimen.

Although disruption of the circadian rhythm had been traditionally considered as a pathological sign, there is an increasing recognition that an existence of internal disorder (or chaos) in the organism's homeostasis is, to some degree, essential to the organism's well being. In this study we explored the effects of rhythm scrambling by exposure to random light/dark (RLD) alternation or by hydrocortisone administration. The variables measured were the toxicity of Adriamycin, Vincristin, Cisplatinum and Cyclophosphamide in C57Bl/6J mice and the survival of EL4 lymphoma-bearing mice, before and after chemotherapy. Rhythm alterations were determined by WBC counts and plasma Alkaline Phosphatase activity. Injections of Adriamycin, Cisplatinum and Vincristin in RLD conditions resulted in a better survival than in control groups of mice kept in LD illumination regimen, although the differences between the groups were significant only for injection of Adriamycin. RLD conditions imposed a "protective" effect on survival of tumor-bearing mice. On the 94th day, 20% of the injected mice in RLD conditions still survived while, there were no survivors beyond 38 days in control group. Chemotherapy had a more prominent beneficial effect on survival in RLD group, as compared to LD group. The injections of hydrocortisone had detrimental effect on survival in both illumination schedules. However, the survival in the RLD group was still better than in the LD group. These experiments indicate that temporal disorganization has beneficial effects on lymphoma-bearing mice and could be used for development of new therapeutic modalities.

Circadian variation of nitric oxide synthase activity in mouse tissue.

Endogenous nitric oxide (NO) is an important mediator in the processes that control biological clocks and circadian rhythms. The present study was designed to elucidate if NO synthase (NOS) activity in the brain, kidney, testis, aorta, and lungs and plasma NOx levels in mice are controlled by an endogenous circadian pacemaker. Male BALB/c mice were exposed to two different lighting regimens of either light-dark 14:10 (LD) or continuous lighting (LL). At nine different equidistant time points (commencing at 09:00h) blood samples and tissues were taken from mice. The plasma and tissue homogenates were used to measure the levels of NO2 + NO3- (NOx) and total protein. The NOx concentrations were determined by a commercial nitric oxide synthase assay kit, and protein content was assessed in each homogenate tissue sample by the Lowry method. Nitric oxide synthase activity was calculated as pmol/mg protein/h. The resulting patterns were analyzed by the single cosinor method for pre-adjusted periods and by curve-fitting programs to elucidate compound rhythmicity. The NOS activity in kidneys of mice exposed to LD exhibited a circadian rhythm, but no rhythmicity was detected in mice exposed to LL. Aortic NOS activity displayed 24h rhythmicity only in LL. Brain, testis, and lung NOS activity and plasma NOx levels displayed 24h rhythms both in LD and LL. Acrophase values of NOS activity in brain, kidney, testis, and lungs were at midnight corresponding to their behavioral activities. Compound rhythms were also detected in many of the examined patterns. The findings suggest that NOS activity in mouse brain, aorta, lung, and testis are regulated by an endogenous clock, while in kidney the rhythm in NOS activity is synchronized by the exogenous signals.

Seasonality of birth and onset of clinical disease in children and adolescents (0-19 years) with type 1 diabetes mellitus in Canterbury, New Zealand.

AIMS: To determine the seasonality of clinical disease onset and month of birth in type 1 diabetes mellitus (DM) in the southern hemisphere. PATIENTS: Two hundred and seventy-five children with type 1 DM in the South Island of New Zealand were studied. The total live births (91,394) of the same period were used as control data. METHODS: Seasonal rhythms were analyzed using the 12 month cosinor method. RESULTS: The month of birth pattern of the patients with DM showed a statistically significant peak (p < 0.01) in summer, whereas the disease onset had a significant peak in winter (p < 0.01), similar to that registered in countries of the northern hemisphere, but in different months of the year. The total live births had no significant rhythm. The different seasonality of birth of the children who subsequently developed type 1 DM from that of the total live births is suggestive of the initiation of the autoimmune process in utero or perinatally.

Right- and left-brain hemisphere. Rhythm in reaction time to light signals is task-load-dependent: age, gender, and handgrip strength rhythm comparisons.

In healthy mature subjects simple reaction time (SRT) to a single light signal (an easy task) is associated with a prominent rhythm with tau = 24 h of dominant (DH) as well as nondominant (NDH) hand performance, while three-choice reaction time (CRT), a complex task, is associated with tau = 24 h of the DH but tau < 24 h of the NDH. The aims of the study were to assess the influence of age and gender on the difference in tau of the NDH and DH, as it relates to the corresponding cortical hemisphere of the brain, in comparison to the rhythm in handgrip strength. Healthy subjects, 9 (5 M and 4 F) adolescents 10-16 yr of age and 15 (8 M and 7 F) adults 18-67 yr of age, active between 08:00 +/- 1 h and 23:00 +/- 1:30 h and free of alcohol, tobacco, and drug consumption volunteered. Data were gathered longitudinally at home and work 4-7 times daily for 11-20 d. At each test time the following variables were assessed: grip strength of both hands (Dynamometer: Colin-Gentile, Paris, France); single reaction time to a yellow signal (SRT); and CRT to randomized yellow, red, or green signal series with varying instruction from test to test (Psycholog-24: Biophyderm, France). Rhythms in the performance in SRT, CRT, and handgrip strength of both DH and NDH were explored. The sleep-wake rhythm was assessed by sleep-logs, and in a subset of 14 subjects it was also assessed by wrist actigraphy (Mini-Motionlogger: AMI, Ardsley NY). Exploration of the prominent period tau of time series was achieved by a special power spectra analysis for unequally spaced data. Cosinor analysis was used to quantify the rhythm amplitude A and rhythm-adjusted mean M of the power spectral analysis determined trial tau. A 24h sleep-wake rhythm was detected in almost all cases. In adults, a prominent tau of 24 h characterized the performance of the easy task by both the DH and NDH. In adults a prominent tau of 24 h was also detected in the complex CRT task performed by the DH, but for the NDH the tau was < 24 h. This phenomenon was not gender-related but was age-related since it was seldom observed in adolescent subjects. Hand-side differences in the grip strength rhythms in the same individuals were detected, the tau being ultradian rather than circadian in adolescent subjects while in mature subjects the tau frequently differed from that of the rhythm in CRT. These findings further support the hypothesis that functional biological clocks exist in both the left and right hemispheres of the human cortex.