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.

Time dependent protection of amifostine from renal and hematopoietic cisplatin induced toxicity.

Efficacy of chemotherapy may be maximized and its toxicity can be minimized if drugs would be administered at specified daily times. The present study was aimed to examine if the protection of amifostine against cisplatin toxicity is time dependent. Amifostine is an organic thiophosphate that protects selectively normal tissues, but not tumors, against the cytotoxicity of DNA binding chemotherapeutic agents such as cisplatin. ICR male mice which were entrained to Light:Dark (L:D) 14:10 were injected (intrapritoneal bolus) for 5 consecutive days with either: cisplatin, cisplatin plus amifostine (administered 30 minutes prior to cisplatin). Injections were given at either 08:00, 13:00, 20:00 or 01:00. Five days later, on day 10, each set of mice was sacrificed (at the same hour corresponds to the injection hour), blood count, blood creatinine and blood urea nitrogen (BUN) were assayed. Cisplatin treated mice exhibited nephrotoxicity, as indicated by increased blood urea nitrogen values and by high blood urea nitrogen to creatinine ratios, as well as myelotoxicity that was indicated by low levels of hemoglobin and platelets. Co-administration of amifostine-cisplatin reversed both, the nephrotoxicity of cisplatin, and its myelosuppressive effects. For BUN, hemoglobin and platelets, maximal protections were observed at 08:00, (p <0.05, p <0.01 and p <0.01 respectively). For BUN/Cr ratio (p <0.05), maximal protections was observed at 13:00. These findings show that amifostine exhibits time dependent protection against cisplatin toxicity and thus it is recommended to use the protector when treatments are given during morning hours. The results also further validate the notion that chronochemotherapy is advantageous at least in reducing drug toxicity and thus should be integrated in the design of clinical protocols.

Gonadotropin releasing hormone (GnRH) and estradiol (E(2)) regulation of cell cycle in gonadotrophs.

The number of pituitary cells, their size, hormonal content and release and response to external cues varies between day and night and during the estrus cycle. Previous studies have demonstrated that pituitary cells proliferate rhythmically and that estradiol (E(2)) is a mitogen of alpha T3 cells. We, therefore, studied the effect of gonadotropin releasing hormone (GnRH) and E(2), on the cell cycle in primary cultures of mouse pituitary cells and in the gonadotroph cell line L beta T2. We found that GnRH and E(2) modulate the cell cycle in a time dependent manner and induce proliferation in cultures of mouse pituitary and L beta T2 cells. GnRH induces proliferation in cells isolated in the morning of the estrus day and increases the number of cells in G2 stage when isolated in noon and evening. However, the transition into the G1 stage is enabled only by co-addition of E(2) and GnRH. GnRH stimulates LH release from L beta T2 cells after 2 days via exocytosis while after 4 days in culture, the increase in LH release may be accounted for by the increase in cell number. E(2) enhanced the GnRH response after 2 days, and abolished it after 4 days in culture. Furthermore, E(2) has no effect on LH release and cell number after 2 days in culture, however, after 4 days in culture, E(2) had no effect on the total amount of LH released but inhibited LH release per cell due to increase in cell number. Our results show that GnRH and E(2) function to shorten the cell cycle and regulate the cell number of each stage of the cell cycle. The effect of GnRH and E(2) on the cell cycle is dependent on the circadian time. This mechanism may serve to modulate the size and function of the pituitary cell population and consequently the function of pituitary gonadotrophs regulating the surge of LH release before ovulation.

Rhythmicity of luteinizing hormone secretion expressed in vitro.

In the present study we explored the possibility that the pituitary functions as an autonomous clock and is capable of generating rhythms of luteinizing hormone (LH) release independently of hypothalamic control. Pituitaries from estrous or diestrous day 1 female mice were perifused separately with Medium-199. Effluent samples were collected at 10-min intervals and assayed for LH levels. Fourier analysis and curve-fit analysis served to elucidate the presence of prominent periods whose significance was then determined by best-fit cosinor. The latter method was used to determine additional parameters for the significant rhythm. All perifused pituitaries exhibited LH release patterns that were composed of significantly long ultradian rhythms (approximately 16 and 8 h, p < 0.001). Continuous stimulation with gonadotropin-releasing hormone (GnRH) or estradiol did not alter the periods of the observed rhythms but affected other rhythm parameters. Gonadotropin-releasing hormone increased the mesor of the rhythm and estradiol increased the amplitude. The results indicate that pituitary gonadotropes are capable of producing rhythms of LH release for a long duration in vitro, in the absence of hypothalamic control. Both GnRH and estradiol affect different rhythm parameters but do not change the periods of these rhythms.

Differences between left- and right-hand reaction time rhythms: indications of shifts in strategies of human brain activity.

Reaction time (RT) measurements serve as quantitative indices for pilots' cognitive processes of the brain. To examine if laterality exists in the brain hemispheres we measured, by the use of a Pilot Evaluation System (PES), right- and left-hand performance rhythms as indicative of RT to audible and visual stimuli. The tests included sets of simple tasks and complex ones to which a secondary task composed of audio signals was added. The accuracy of recorded reaction time was 27 ms. Seven right-handed males, 27-42 years of age, experienced with the PES flight simulator, were tested every 2 h, nine times daily (starting at 08:00 h) during 3 consecutive days. The results indicated that for simple tasks, the 24 h period of RT rhythm is either exclusive or prominent for both hands. For complex tasks the prominent period of RT is 24 h for the right (dominant) hand and 8 h for the left (non-dominant) hand (right-hand 24 h period Fstat = 140, r2 = 0.62 and 8 h period Fstat = 25, r2 = 0.22; left-hand 24 h period Fstat = 44, r2 = 0.34 and 8 h period Fstat = 100, r2 = 0.54). The findings suggest that a laterality exists in the brain hemispheres with regard to differences in rhythm periodicities. The expression of this laterality is dependent on the task-load level and points to a strategy of linkage and integrity in brain activity.

Time dependency of hematopoietic growth factor coupled to chronotoxicity of carboplatin.

A growing body of data suggests that cancer therapy may be improved and toxicity reduced by administration of antineoplastic agents and cytokines at carefully selected times of the day. The time-dependent effects of each of the drugs have been documented, but not their mutual time dependencies. In the present studies we sought to determine the best time for granulocyte colony-stimulating factor (G-CSF) administration after carboplatin treatment. Carboplatin was injected in different groups of ICR mice at four different circadian stages for 5 consecutive days. Mice were synchronized with an alternation of 12 h of light (from 6:00 a.m. to 6:00 p.m.) and 12 h of darkness. After the last injection, peripheral WBCs of three mice from each group were counted every 4 h over a 24-h period. Bone marrow toxicity was estimated with the mean 24-h WBC count. The most severe leukopenia occurred in the group injected at 3:00 p.m. - 9 h after light onset. The second set of experiments evaluated the time-dependent effect of G-CSF when singly injected or given after carboplatin injections for 5 days only at 3:00 p.m. G-CSF was injected into various groups on days 8 and 9 at the same four different circadian stages. On the 10th day after the first injection, peripheral WBCs of three mice from each group were counted every 4 h over a 24-h period. Time-dependent effects were observed when G-CSF was injected as a single agent. When G-CSF was given at various times to the group with the most severe carboplatin-induced leukopenia, peripheral WBC count recovery was monitored at all injection times; it reached its highest level (exceeding even that of the control) when G-CSF was injected at 3:00 a.m. Dosing times of both chemotherapy and growth factor are relevant for optimization of carboplatin's hematologic tolerability.

Herpetic eye attacks: variability of circannual rhythms.

BACKGROUND: The issue of seasonal variation of herpetic ocular infections is still controversial. This study was designed to examine whether this variation exists and can be defined as a significant circannual rhythm. METHODS: The patterns of recurrent attacks were monitored in 541 patients over a period of 15 years. Rhythm parameters were analysed according to age, sex, and clinical signs. RESULTS: The majority of herpetic eye attacks exhibited the highest peak in January (p < 0.04), except in the group of atopic children where the incidence of the disease peaked in September (p < 0.05). Among the various clinical forms, significant circannual periodicities were found only in the occurrence of epithelial herpetic keratitis (p < 0.03). The rhythms were detected among males (p < 0.03) but not among females. No direct correlation was demonstrated between the presence of the rhythms and the triggering effect of upper respiratory tract infections. CONCLUSIONS: Chronoepidemiological evaluation of individual reactivation patterns may be beneficial to certain patients and contribute to the optimisation of the treatment when prophylaxis is considered.

Genetic control of biological rhythms: independent expression of each rhythm parameter.

Biological rhythm whose pattern approaches a sinusoidal curve can be characterized by four parameters: Period, Mesor, Acrophase and Amplitude. Inheritance patterns of these parameters were assessed by monitoring White Blood Cell (WBC) count rhythms of C57BL/6J mice, BALB/c mice and the F1 progeny prior and after injection of Cisplatinum at either 0900, 1500, 2100, or 0300. All untreated mice exhibited similar WBC rhythms while a variety of rhythm phenotypes was revealed among the injected mice. Analyses of the variety in the parental strains and F1 progeny suggested that each of the rhythm parameters is independently controlled.

Strain dependent response of circadian rhythms during exposure to continuous illumination.

Enzymes activities were measured, at three hours intervals, during 30 hours, in various tissues of C57BL/6J and A/J male mice. The measurements, were carried out on mice which were exposed for two, five and twenty one days to continuous illumination. Identical measurements were performed also on mice which were kept in alternating 14 hours light: 10 hours dark. Activity patterns of each group were analysed to test the presence, or absence, of rhythm characteristics. The results of the experiments with C57BL/6J have been previously reported. The comparison of the results, which were obtained from the two strains revealed that under exposure to alternating light: dark conditions all activity patterns exhibited a significant circadian rhythm. Except for one enzyme (thymus GAPD), the times of peak activity (acrophase) were identical for all other examined enzymes, in both strains. On the other hand when the two strains were exposed to continuous illumination they differed in their response to the effect of continuous light. The activity of the same enzyme exhibited different periodicity and/or different acrophase in each of the two strains. This variability reflects the existence of genetic differences, between the strains in the free running behavior of these enzymes' activity rhythms.

Gender-dependent differences in biological rhythms of mice.

The advantage of a variable's rhythm resides in its optimal time-phasing. This implies that, for a given function, members of a species will strive to exhibit identical time-phasing namely, their inter-individual genetic differences will be masked. To examine the generality of this assumption we explored if inbred mice exhibit gender dependent differences in rhythm parameters of biochemical variables. Male and female mice, entrained by exposure to 12:12 light:dark illumination were sacrificed, every 3 hours over a 27 hours period. Activities of creatine-phosphokinase (CK) and alkaline- phosphatase (AP), white blood cell (WBC) counts and urea nitrogen (UN) concentration were determined at each time point. For each significant rhythm four parameters were computed: period, acrophase, mesor and amplitude. In addition two derived parameters were also calculated: relative-amplitude (RA) and the rate of change in RA (CRA) which provide information about the slope and width of the peak. Patterns of most variables exhibited a compound rhythm containing two significant periodicities. Gender dependent differences were documented in the parameters of most rhythms indicating that the genetic and physiological differences limit to a certain extent the phasing ability of the entraining signals and point to an independent control of each of the rhythm parameters.

Compound structure of rodents activity rhythm.

In the present study an attempt to determine the fine structure of rodents activity rhythm was carried out. To eliminate masking effects which are produced by the active influence of the monitoring system on the behavior of the animal (e.g., running wheel) we designed a passive infra red detection system. Rats were exposed to light-dark 12:12 [LD(12:12)] cycles and mice to LD(12:12), LD(8:16) and LD(16:8) cycles. Multiwave patterns of activity were observed in both rodents genera which differ from each other in the number of activity bouts and the periods of the activity rhythm components. In LD(12:12), rats exhibited 2 bouts of activity and 1 bout of rest which were attributed to the presence of 24 and 8 h components. Mice, exposed to the three varying ratios of L to D regimens exhibited 3 bouts of activity and one bout of rest which were attributed to the presence of 24 and 6 h components. The relation of the compound structure to the 24 h rhythm is discussed.

Circadian variation of nystagmus in healthy and sick subjects.

Rhythmic activity of various biological systems and functions have been investigated extensively. However, information about diurnal variations of the vestibular system is scarce. In the present preliminary report, diurnal variations of the spontaneous and induced nystagmus in healthy subjects and in patients with peripheral vestibular disorders have been assessed. Circadian rhythms of the vestibular responses were demonstrated. In some variables significant differences have been found between health and sick examinees. These differences may have practical clinical implications on diagnosis and treatment.

Interindividual differences in chronopharmacologic effects of drugs: a background for individualization of chronotherapy.

In order to optimize chronotherapeutic schedules (designs), we examined the interindividual differences in chronopharmacologic effects of drugs with consideration of the following three factors: (a) inherited factors of direct relevance to chronopharmacology (genetic variability, gender-related differences) as well as age-related differences; (b) interindividual difference in chronoeffectiveness related to disease (e.g., various types and stages of cancer, affective disorders, etc.) as well as to drug-dependent alteration (phase shifts, distortion) of biological rhythms; and (c) means to solve problems resulting from the need of individualization in chronotherapy. These involve the use of circadian marker rhythms (MR) whose characteristics (peak or trough time, amplitude, etc.) can be precisely quantified and thus are applicable as a reference system for physiologic, pathologic, pharmacologic, and therapeutic uses. The MR has to be specific and pertinent and must be easily monitored and documented. This approach can be further advanced by the use of a battery of MRs rather than a single MR. Other suggested means relate to the fact that chronobiotics (agents capable of influencing parameters of a set of biological rhythms) should be considered (e.g., corticoids and adrenocorticotropic hormone) and/or to the subject's synchronization should be enforced by "conventional" zeitgebers (e.g., bright light, physical activity).

From ultradian to infradian rhythms: LH release patterns in vitro.

In the present study, we examined in vitro luteinizing hormone (LH) release patterns from pituitaries and from pituitary cell cultures (3 and 7 days in culture) to elucidate the endogenous period generated by the gonadotroph cell population and to evaluate the relationship between the basic period generated at the cellular level and the output pattern observed at the organ level. In addition, we examined the effect of photic environmental signals perceived by the animals on LH release patterns from pituitaries in vitro. When the animals were exposed to circadian photoperiodic signals, the in vitro LH release pattern from the pituitaries exhibited ultradian, circadian, and infradian frequencies. When the animals were exposed to continuous illumination, the in vitro patterns exhibited only ultradian and infradian frequencies. Furthermore, free running is a process, not a state. This process is driven by a change in the relative dominance of different frequencies that construct the pattern without changing the basic period length. Evaluation of the relative dominance of the different frequencies that construct the pattern indicates that, although infradian oscillators may take part in shaping the output pattern, the basic rhythm generated by the pituitary cells is in the ultradian domain. The results obtained from the examined system suggest that an endogenous oscillator is a cellular entity with ultradian periodicity, and that the rhythmic output of many biological variables is structured by various ultradian components that construct the circadian and infradian output rhythms.

Interindividual differences in the flexibility of human temporal organization: pertinence to jet lag and shiftwork.

Interindividual variability in the human temporal structure is seldom taken into account, especially in studies devoted to the effects of shiftwork and jet lag. The understated postulate is that humans can be treated as a pure strain species. This paper reviews some facts and concepts with special reference to interindividual changes in the rhythm period tau and the resulting dyschronism. The following points are addressed. (1) Subjects and methods (importance of longitudinal field studies on shift workers). (2) Criteria for tolerance to shiftwork and jet lag. (3) Interindividual differences and shiftwork problems (subject type; the association between good shiftwork tolerance and stable temporal structure; dychronism with tau s differing from 24h and from variable to variable. (4) The genetic background of circadian dyschronism. The Dian-circadian genetic model of biological rhythms. It allows understanding of one's susceptibility to dyschronism, which was actually observed in approximately equal to 30% of subjects studied longitudinally. (5) Practical implications of interindividual differences (dissociate problems of passengers after a transmeridian flight-who have to adjust their temporal structure to local time-from problems of shiftworkers-who need to prevent alteration of their temporal structure; the advantage for the latter of participating in a rapid rotation system rather than a weekly rotation; emphasis that the suitability of a given subject for a given shiftworking condition is likely to be estimated only after a trial span of time including longitudinal study of a set of rhythms.

Circadian pattern of simulated flight performance of pilots is derived from ultradian components.

Studies suggest some physiologic, cognitive, and behavioral 24h rhythms are generated by cyclic components that are shorter in period than circadian. The aim of this study was (1) to examine the hypothesis that 24h human performance rhythms arise from the integration of high-frequency endogenous components and (2) to quantify the contribution of each higher frequency component to the phenotype of the rhythm. We monitored the performance of 9 experienced pilots by employing an array of cognitive-based tests conducted in a flight simulator so that, over the 6-day experiment, data were obtained for each 2h interval of the 24h. The activity-rest schedule of the subjects, no matter the exact clock time schedule of sleep and activity, always consisted of 14h activity (when they carried out regular professional duties) and 10h rest, with at least 8h of sleep. The simulated combat scenarios consisted of simple and complex tasks associated with target interception, aircraft maneuvering, and target shooting and downing. The results yielded two indices: the number of prominent periodicities in the time series and the relative magnitude of the amplitude of each relative to the construction of the composite 24h waveform. Three cyclic components (8h, 12h, and 24h) composed the observed 24h performance pattern. The dominant period and acrophase (peak time) of the compound output rhythm were determined by the interplay between the amplitudes of the various individual ultradian components. Task complexity (workload) increases the expression of the ultradian entities in the 24h pattern. We constructed a model composed of the multiple ultradian components; the composite output defined a "time span" (of 2h-4h duration) as opposed to an exact "time point" of high and low performance, endowing elevated functional capability.

Human circadian time structure in subjects of different gender and age.

Most human variables exhibit rhythms with an about 24 hour (circadian) period. Each rhythm can be characterized by its acrophase (calculated peak time of the cosine curve best fitting to the data), its amplitude and rhythm adjusted mean (MESOR). The sequential array of the rhythms' acrophases represents the temporal order of the human time structure. In the present work we used circadian rhythms of 24 chemical and 15 hormonal variables extracted from published studies which were done in a defined area of southeastern Europe (Romania). All studies had a comparable experimental design and were analyzed biochemically and statistically in the same laboratory. The acrophases of these rhythms obtained from both genders of different age groups (from the 2nd to the 9th decade of age) were subjected to multiple correlation test, cluster and principal coordinates analyses. The results show that the temporal order is affected both by gender and age, and evaluate the degree of the effect, offer a "chronbiologic fingerprint" for the examined groups and assist in dissecting rhythm variability among populations.

Temporal pattern of LH secretion: regulation by multiple ultradian oscillators versus a single circadian oscillator.

The possibility that the 24h rhythm output is the composite expression of ultradian oscillators of varying periodicities was examined by assessing the effect of external continuously or pulsed (20-minute) Gonadotropin-releasing hormone (GnRH) infusions on in vitro luteinizing hormone (LH) release patterns from female mouse pituitaries during 38h study spans. Applying stepwise analyses (spectral, cosine fit, best-fit curve, and peak detection analyses) revealed the waveform shape of LH release output patterns over time is composed of several ultradian oscillations of different periods. The results further substantiated previous observations indicating the pituitary functions as an autonomous clock. The GnRH oscillator functions as a pulse generator and amplitude regulator, but it is not the oscillator that drives the ultradian LH release rhythms. At different stages of the estrus cycle, the effect of GnRH on the expression of ultradian periodicities varies, resulting in the modification of their amplitudes but not their periods. The functional output from the system of ultradian oscillators may superimpose a "circadian or infradian phenotype" on the observed secretion pattern. An "amplitude control" hypothesis is proposed: The temporal pattern of LH release is governed by several oscillators that function in conjunction with one another and are regulated by an amplitude-controlled mechanism. Simulated models show that such a mechanism results in better adaptive response to environmental requirements than does a single circadian oscillator.

Circadian rhythms of enzymes' activity in mice tissues during exposure to continuous illumination.

Activity rhythms of enzymes were determined in various tissues of C57BL/6J male mice. The determinations were carried out on mice which were kept in 14 hr light: 10 hr dark regimen, and on day 2, day 5 and day 21 during exposure to continuous illumination. Locomotor activity rhythms were followed in light: dark and up to the seventh day in constant light. All the activities exhibited a significant circadian rhythm in the light: dark regimen. During the exposure to continuous illumination, the locomotor activity exhibit a free running circadian rhythm with a consistent 24 hr and 40 min, major period component. At the same time recording the rhythms of enzyme activity; enzymes exhibited various formats of response which differed from those of the locomotor activity. The results suggest that rhythms of enzyme activity, as well as the desynchronization of the rhythms, are not enzyme specific.

Placebo effect on the circadian rhythm period tau of temperature and hand-grip strength rhythms: interindividual and gender-related difference.

Two different medications, one assumed to be a tranquilizer and the other an antifatigue agent, were tested. Both were found to be ineffective and thus were viewed as placebos and named P1 and P2. The effect of P1 and P2 on the circadian rhythms of a set of variables (e.g., sleep/wake, oral temperature and grip strength of both hands) were monitored by five to 10 measurements per day over three consecutive 8- to 10-day spans. The first documented span was a control (no medication), and the second and third spans (in randomized order) were under P1 and P2. Healthy subjects volunteered for the studies: nine men and seven women (median age 28 years) in study 1 and 12 men and 12 women (median age 36 years) in study 2. They were synchronized with diurnal activity from 07:00 h (+/- 30 min) to 00:00 h (+/- 1 h) and nocturnal rest. De Prins' method was used to obtain the prominent period tau in each (control, P1, and P2) individual time series. The chi 2 test was used to test group and subgroup differences. All 40 subjects exhibited a significant sleep/wake rhythm with a tau = 24 h in control, P1, and P2 series. During the control span a gender-related statistically significant difference was observed: fewer men than women exhibited a temperature rhythm desynchronized from 24 h. In addition, more women than men had a tau < 24 h during control. The gender-related difference was obliterated by placebos. Similarly desynchronized circadian rhythms of left and right hand-grip strength were observed in both men and women during the control span, which were all obliterated by placebo but only in men. Results are discussed with regard to a genetic model of human dyschronism as proposed by the authors.