The relative Zeitgeber strength of lights-on and lights-off is changed in old mice

The daily activity pattern of old mice is characterized by a decreased amplitude, a phase advance, and less stable relationship between lights-off and the onset of the main activity maximum. When analyzing the possible causes of these changes, it must be remembered that the activity rhythm of laboratory mice is bimodal, with a main peak in the first half of the dark time and a secondary one shortly after lights-on. Thus it seems to be controlled by at least two circadian oscillators--an "evening oscillator" coupled more strongly to lights-off and a "morning oscillator" coupled to lights-on--though both oscillators are also coupled to each other. The objective of the present paper was to investigate the putative changes in the strength of these couplings in HaZ:ICR mice of different ages (adult animals of 20 weeks, n = 12; old mice of 72 and 91 weeks of age, n = 6 each) and kept in a 24 h LD-cycle with a gradually reduced light:dark ratio. In adult mice, lengthening the dark time caused the onset of the main maximum of activity to be delayed in relation to the time of lights-off, while the morning maximum of activity was advanced in relation to lights-on. On average, the sizes of the advance and the delay were equal. As a consequence, the activity pattern did not shift in relation to the middle of the dark time. Lengthening the dark time resulted in a bigger (on average, 1.5 h) difference between the evening and morning activity onsets. Under short photoperiods (< or = 2 h of light) the activity rhythm started to free run, and the difference between evening and morning activity onsets decreased again. The changes obtained in senile mice were similar. However, the limits of entrainment were reached with longer photoperiods compared to adult animals. Also, the phase delay of the activity onset in the evening was much less, nearly zero. As a consequence, the activity pattern as whole phase-advanced in relation to the middle of the dark time. A model was proposed in which lights-off triggers advances of the "evening oscillator," lights-on delays the "morning oscillator," and the two oscillators are coupled with each other. Though it was probably the case, decreased coupling strengths could not be shown with the present experimental approach. However, it was clearly evident that, with increasing age, the advancing effect of lights-off exceeded the delaying effect of lights-on.     

Possible evidence for morning and evening oscillators in Drosophila melanogaster populations selected for early and late adult emergence

In this paper, we report the results of our study aimed at a systematic analysis of the circadian phenotypes of fruit flies Drosophila melanogaster selected for early and late adult emergence, in light of the "morning and evening oscillator" (M and E) model for circadian clocks. We monitored adult emergence and activity/rest rhythms in these flies under light/dark (LD) cycles with short (8:16 h), normal (12:12 h) and long (16:8 h) photoperiods, as well as under constant darkness (DD). Across all the three LD cycles, the early populations displayed a morning phenotype with peak of emergence and activity occurring earlier than the controls and greater anticipation to "lights-on" and weak anticipation to "lights-off", while the late populations showed an evening phenotype with peak of emergence and activity occurring later than the controls and greater anticipation to lights-off and weak anticipation to lights-on. The gate of adult emergence and duration of activity in the early populations was narrower than the controls, while those of the late populations were wider than the controls. In addition, the circadian periodicities of adult emergence and activity/rest rhythms of the early flies were significantly shorter than the controls, while those of the late flies were significantly longer than the controls. In summary, the circadian phenotypes indicate that the early populations have evolved a dominant M oscillator, while the late populations have evolved a dominant E oscillator, thus providing an empirical support for the M and E model in Drosophila.     

Temperature profiles, and the effect of sleep on them, in relation to morningness-eveningness in healthy female subjects

There were 15 healthy female subjects, differing in their position on the “morningness-eveningness” scale, studied for 7 consecutive days, first while living a sedentary lifestyle and sleeping between midnight and 08:00 and then while undergoing a “constant routine.” Rectal temperature was measured at regular intervals throughout this time, and the results were subjected to cosinor analysis both before and after “purification” for the effects of physical activity. Results showed that there was a phase difference in the circadian rhythm of core temperature that was associated with the morningness score, with calculations that “morning types” would be phased earlier than “evening types” by up to about 3h. This difference in phase (which was also statistically significant when the group was divided by a median split into a “morning group” and an “evening group”) could not be attributed to effects of waking activity and existed in spite of the subjects keeping the same sleep-wake schedule. Moreover, it persisted when the subjects' data had been purified and when the data were obtained from the constant routine. That is, there was an endogenous component to this difference in phase of the core temperature. The morning group also showed a greater fall of core temperature during sleep; this was assessed in two ways, the main one being a comparison of constant routine and nychthemeral data sets after correction for any effects of activity. Even though the morning group was sleeping at a later phase of their circadian temperature rhythm than was the evening group, neither group showed a fall of temperature due to sleep that varied with time elapsed since the temperature acrophase. It is concluded that another factor that differs between morning and evening types is responsible for this difference. (Chronobiology International, 18(2), 227-247, 2001)     

Differential control of morning and evening components in the activity rhythm of Drosophila melanogaster--sex-specific differences suggest a different quality of activity

The rhythms of locomotor activity of male and virgin or mated female flies were compared in the Drosophila melanogaster wild-type strains CantonS, Berlin, and OregonR. Under light-dark conditions, most flies showed a bimodal activity pattern with a morning peak around lights-on and an evening peak before lights-off. For all strains, a distinct sexual dimorphism was observed in the phase of the morning peak. Males had a significantly earlier morning peak than females and consequently a larger phase angle between morning and evening peak (psi(m, e)). Under constant dark conditions, the morning component merged with the evening component to a unimodal activity band in about half of the flies. In those flies who maintained bimodality, the sex-specific difference in psi(m, e) disappeared. Other sex-specific differences were now apparent: Males showed a shorter free-running period than females, and in two of the three strains, females were more active than males. Morning and evening components seem to contribute to the free-running period. Spontaneous or externally provoked change in psi(m, e) were correlated with period changes. In some flies, the morning and the evening components showed splitting, indicating that they are the output of two different oscillators. The sexual dimorphism in the phase of the morning peak under LD-conditions suggests that the function of activity during morning and evening peak might be different, for example, during the morning peak, males are active to find females. Overall, the results underline the multioscillatory nature of Drosophila's circadian system.     

Entrainment analysis of the circadian locomotor activity rhythm in specimens of the crayfish, Faxonella clypeata, having activity peaks at different times of the solar day.

Locomotor activity rhythms in the crayfish, Faxonella clypeata, were recorded under conditions of controlled light and temperature. In LD 12:12, dark active rhythms with a major activity onset at lights-off, and bimodally active rhythms with onsets at both lights-on and lights-off were recorded. In DD, most of the LD dark active rhythms retained the lights-off activity onset. However, among the majority of the LD bimodally active crayfish, only the lights-on activity peak persisted in DD. A small number of the LD bimodal rhythms remained bimodal in DD. DD records revealed free-running period lengths both greater and less than 24 hrs. An hypothesis stating that the two recorded LD rhythms represent two basically different rhythmic types is presented. The dark active crayfish appear to entrain to the lights-off position, but the bimodally rhythmic crayfish appear to entrain to the lights-on position.     

Different mechanisms of phase delays and phase advances of the circadian rhythm in rat pineal N-acetyltransferase activity

The circadian rhythm in rat pineal N-acetyltransferase (NAT) activity, which drives the rhythm in melatonin production, is controlled by a pacemaker located in the suprachiasmatic nucleus of the hypothalamus. As the NAT rhythm has two well-defined phase markers--namely, the time of the evening activity rise and of the morning decline--it is suitable for studies of the entrainment of the pacemaker by environmental light. Phase delays of the NAT rhythm proceed more rapidly than phase advances. One day after a brief light pulse applied before midnight, or after a delay in evening lights-off, or a delay of a light-dark (LD) cycle, phase delays of the evening NAT rise result in almost corresponding delays of the morning NAT decline. Consequently, the NAT rhythm is phase-shifted, but its pattern does not change. One day after a brief light pulse applied past midnight, or after bringing forward morning lights-on, or after an advance of an LD cycle, the morning NAT decline is phase-advanced, but the evening rise is not phase-advanced at all or may even by phase-delayed. Consequently, the phase relationship between the evening NAT activity onset and the morning offset may be compressed considerably, and it may take several transient cycles before phase advances of the morning NAT decline are followed by corresponding advances of the evening NAT rise. Due to the phase-delaying effect of evening light on the NAT rise and to the phase-advancing effect of morning light on the NAT decline, the phase relationship between the NAT rise and the decline is compressed on long days and decompressed on short days. Different phase shifts of the evening NAT rise and of the morning decline, even in opposite directions, are consistent with the hypothesis of a complex, two-component (evening-morning, or E-M) pacemaker controlling the NAT rhythm. As the E-M phase relationship determines duration of the high night melatonin production, and the duration of the nocturnal melatonin pulse may convey information on daylength, the data are consistent with the internal coincidence model for photoperiodic time measurement.     

The Difference Between Activity When in Bed and Out of Bed. II. Subjects on 27-Hour “Days”

Nine healthy subjects have been studied while exposed to the normal alternation of light and dark, but with their sleep and activity pattern adjusted to a 27-h “day” for 17 imposed “days.” Rectal temperature showed clearly the competing influences of 27-h and 24-h components, and these were separated by the method of “purification.” The method indicated that the endogenous component had a constant amplitude throughout the experiment and remained entrained to solar (24-h) time; by contrast, the exogenous component followed the imposed 27-h “day” and increased rectal temperature in proportion to the amount of subjects' activity. Wrist movement was used to assess activity while in bed (attempting sleep) and out of bed (when naps were forbidden). While these results confirmed adherence of the subjects to the imposed 27-h “days,” they also showed that the dichotomy between “out of bed” activity and “in bed” inactivity depended on the phase relationship between endogenous (24h) and exogenous (27h) components. Thus, the dichotomy was highest and was equal to that during control days (with a conventional 24-h life-style) when the two components were in phase and lowest when the solar and imposed day were in antiphase. This was due to changes in activity, both during time spent in bed and out of bed.

We confirm that this protocol can produce valuable information about the properties of the circadian system in humans and the value of the process of purification of temperature data. We have established also that the very simple and noninvasive measurement of wrist movement, coupled with its use to calculate dichotomy indices, provides valuable information that both confirms and extends the results obtained from the more conventional (butalso more invasive) measurement of rectal temperature.     

Parametric and nonparametric entrainment of circadian locomotor rhythm in the Japanese honeybee Apis cerana japonica

The circadian rhythm of locomotor activity in the Japanese honeybee Apis cerana japonica was studied to determine the involvement of parametric and/or nonparametric entrainment. The rhythm was entrained to a skeleton photoperiod in which a 1-h first light pulse was imposed in the morning along with a second light pulse in the evening, as well as to a complete photoperiodic regime (LD 12:12). However, the timing of peak activity relative to the lights-off in the evening in the skeleton photoperiod was earlier than that in the complete photoperiod. A single daily light pulse in the evening entrained the rhythm, whereas a daily light pulse in the morning allowed free-running as in constant darkness. The free-running period (τ) of locomotor activity in constant light became longer as the light intensity increased. A Winfree's type I phase response curve of the locomotor activity rhythm was obtained using a single 1-h light pulse. The results suggest that both parametric and nonparametric entrainment are involved in the circadian rhythm of individual locomotor activity in this honeybee.     

Effects of photoperiod and aging on locomotor activity rhythms in the onion fly,Delia antiqua

At photoperiods longer than 8h per 24h, adults of the day-active onion fly Delia antiqua showed a major peak of locomotor activity in the late photophase and also bursts of activity induced by lights-on or lights-off. At shorter photoperiods the activity peaks fused. After transfer from long photoperiods to constant darkness (DD), the rhythm free-ran, but only the major peak persisted. This suggests that only the major peak is controlled by the circadian pacemaker. At long photoperiods, the daily phase of the major peak occurred progressively later with age. As a result, the activity at short photoperiods often shifted from photophase to scotophase in old flies. The free-running period (tau) also changed with age; tau was shorter than 24h until 14-20 days after eclosion and thereafter became longer, but a few individuals repeated changes in tau. The phase delay of locomotor activity with age in D. antiqua would be attributable to the increase in tau.     

Circadian Locomotor Activity Under Artificial Light in the Freshwater Crab Pseudothelphusa americana

Long-term recordings of locomotor activity were obtained from intact freshwater crabs, Pseudothelphusa americana in constant darkness (DD), constant light (LL) and different light-dark (LD) protocols. Bimodal rhythms were typically observed in this crab when subjected to DD or LD, with bouts of activity anticipating lights-on and lights-off, respectively. Freerunning circadian rhythms were expressed in both DD and LL for longer than 30 days. In DD, we observed that some animals presented different period lengths for each activity component. During LL, activity was primarily unimodal, however spontaneous splitting of the rhythms were observed in some animals. When activity was recorded under artificial long days, the morning bouts maintained their phase relationship but the evening bouts changed their phase relationship with the Zeitgeber. Our results indicate that, bimodal locomotor activity rhythm in the crab Pseudothelphusa americana is variable among organisms. The characteristics of phase relationship with LD and responses to LL for morning and evening bouts, suggest that, locomotor activity could be driven by multiple oscillators, and that coupling between these oscillators may be regulated by light.     

Circadian Locomotor Activity Under Artificial Light in the Freshwater Crab Pseudothelphusa americana

Long-term recordings of locomotor activity were obtained from intact freshwater crabs, Pseudothelphusa americana in constant darkness (DD), constant light (LL) and different light-dark (LD) protocols. Bimodal rhythms were typically observed in this crab when subjected to DD or LD, with bouts of activity anticipating lights-on and lights-off, respectively. Freerunning circadian rhythms were expressed in both DD and LL for longer than 30 days. In DD, we observed that some animals presented different period lengths for each activity component. During LL, activity was primarily unimodal, however spontaneous splitting of the rhythms were observed in some animals. When activity was recorded under artificial long days, the morning bouts maintained their phase relationship but the evening bouts changed their phase relationship with the Zeitgeber. Our results indicate that, bimodal locomotor activity rhythm in the crab Pseudothelphusa americana is variable among organisms. The characteristics of phase relationship with LD and responses to LL for morning and evening bouts, suggest that, locomotor activity could be driven by multiple oscillators, and that coupling between these oscillators may be regulated by light.     

Rhythms in Fos expression in brain areas related to the sleep-wake cycle in the diurnal Arvicanthis niloticus

Most mammals show daily rhythms in sleep and wakefulness controlled by the primary circadian pacemaker, the suprachiasmatic nucleus (SCN). Regardless of whether a species is diurnal or nocturnal, neural activity in the SCN and expression of the immediate-early gene product Fos increases during the light phase of the cycle. This study investigated daily patterns of Fos expression in brain areas outside the SCN in the diurnal rodent Arvicanthis niloticus. We specifically focused on regions related to sleep and arousal in animals kept on a 12:12-h light-dark cycle and killed at 1 and 5 h after both lights-on and lights-off. The ventrolateral preoptic area (VLPO), which contained cells immunopositive for galanin, showed a rhythm in Fos expression with a peak at zeitgeber time (ZT) 17 (with lights-on at ZT 0). Fos expression in the paraventricular thalamic nucleus (PVT) increased during the morning (ZT 1) but not the evening activity peak of these animals. No rhythm in Fos expression was found in the centromedial thalamic nucleus (CMT), but Fos expression in the CMT and PVT was positively correlated. A rhythm in Fos expression in the ventral tuberomammillary nucleus (VTM) was 180 degrees out of phase with the rhythm in the VLPO. Furthermore, Fos production in histamine-immunoreactive neurons of the VTM cells increased at the light-dark transitions when A. niloticus show peaks of activity. The difference in the timing of the sleep-wake cycle in diurnal and nocturnal mammals may be due to changes in the daily pattern of activity in brain regions important in sleep and wakefulness such as the VLPO and the VTM.     

per mRNA cycling is locked to lights-off under photoperiodic conditions that support circadian feedback loop function.

Circadian fluctuations in per mRNA and protein are central to the operation of a negative feedback loop that is necessary for setting the free-running period and for entraining the circadian oscillator to light-dark cycles. In this study, per mRNA cycling and locomotor activity rhythms were measured under different light and dark cycling regimes to determine how photoperiods affect the molecular feedback loop and circadian behavior, respectively. These experiments reveal that per mRNA peaks in abundance 4 h after lights-off in photoperiods of < or = 16 h, that, phase shifts in per mRNA cycling and behavioral rhythmicity occur rapidly after flies are transferred from one photoperiod to another, and that photoperiods longer than 20 h abolish locomotor activity rhythms and leave per mRNA at a median constitutive level. These results indicate that the per feedback loop uses lights-off as a phase reference point and suggest (along with previous findings for per01 and tim01) that per mRNA cycling is not regulated via simple negative feedback from the per protein.     

Two oscillators might control the locomotor activity rhythm of the high-altitude Himalayan strain of Drosophila helvetica

The properties of the pacemaker controlling the adult locomotor activity rhythm of the high-altitude Himalayan (haH) strain (Hemkund Sahib, 4121 m above sea level) of Drosophila helvetica are strikingly different from those of the low-altitude Himalayan (laH) strain (Birahi, 1132 m above sea level) of the same species. The haH strain has a unimodal activity pattern with a delayed peak occurring about 4.5 h after lights-on of the entraining light-dark (LD) cycle, while the laH strain has a bimodal activity pattern with the morning and evening peaks. It is rather unusual for a wild type strain of any Drosophila species to have a unimodal activity pattern during entrainment as observed in the haH strain. The single activity peak of the haH strain is regarded as a consequence of delayed morning peak merging with the evening one. Three experiments were performed to test this hypothesis. The first experiment examined whether the single activity peak could be dissociated into two components by LD cycles in which photoperiods varied from 10 to 16 h per 24 h. The haH strain again exhibited a unimodal activity pattern with a delayed peak in 10, 12, and 14 h photoperiods but a bimodal activity pattern in 16 h photoperiod. The laH strain had bimodality in 10 and 12 h photoperiods, unimodality in a 14 h photoperiod, but complete arrhythmicity in a 16 h photoperiod. In the second experiment, the haH flies were transferred from LD 16:8 to LL at 5 lux to confirm whether the bimodality of this strain in LD 16:8 cycles was not the result of masking by the long photoperiod of 16 h. Bimodality of the haH strain persisted in LL too; moreover, the morning component free-ran with period (tau) <24 h, while the evening component free-ran with tau>24 h. The third experiment examined the LL-induced splitting of activity peak of the haH strain. Flies were transferred from LD 12:12 cycles to LL at 0, 1, 5, and 15 lux. The haH strain was rhythmic in LL at 0 and 1 lux with a unimodal activity pattern. It was also rhythmic in LL at 5 lux, but the single activity peak was split into two discrete components; the morning component free-ran with tau<24 h, while the evening component free-ran with tau>24 h. This strain, however, was completely arrhythmic in LL at 15 lux. The laH strain was uniformly arrhythmic in LL at all levels of light intensity. These results suggest that the single but late activity component of the haH strain during entrainment appears to be the consequence of merging the delayed morning peak with the evening one as an adaptation to the environmental conditions at the altitude of origin of this strain, where these flies begin activity in the forenoon owing to non-permissible low temperature in the morning.     

The periodicity of pheromone release and male responsiveness in the artichoke plume moth, Platyptilia carduidactyla

ABSTRACT. The periodicity of calling by female Platyptilia carduidactyla (Riley) is influenced by temperature and photoperiod. Both diurnal and nocturnal temperature changes shift the period of nocturnal calling, but in fundamentally different ways; lower daytime temperatures delay activity, while lower night temperatures result in earlier activity. Lengthening photoperiods result in activity earlier in the scotophase, but the period of activity does not maintain a constant phase relationship with either lights-on or lights-off. The periodicity of the behavioural response of males parallels that of calling by females, resulting in coordination of reproductive activities in the field. The result of these responses to environmental stimuli is that mating activity can occur throughout the year in coastal California.     

ESCODD: European Standards Committee on Oxidative DNA Damage

“There can be no more important goal in a clinical laboratory than that of ensuring that the results produced have the precision and accuracy necessary to aid diagnosis and treatment.”

T.P. Whitehead 1976     

Entrainment by different environmental stimuli in the frugivorous bats from the Lonar crater

The timings of emergence at dusk and return at dawn of a colony of frugivorous bats Rousettus leschenaulti, roosting in a temple ruin of the Lonar crater (19.97°N, 76.52°E), were investigated at 10-day intervals for one year. The onset of emergence occurred about 18 min after sunset throughout the year irrespective of the prevailing light intensity or temperature but the end of returning activity occurred at a fixed light intensity of about 4 lux irrespective of the time of sunrise or prevailing temperature. Apparently, the evening oscillator controlling the onset of emergence was set by the sunset, i.e. the lights-off stimulus of the natural light - dark cycles, while the morning oscillator controlling the end of activity was set by a certain invariant threshold intensity of the dawn twilight.     

Prominent Orcadian Absorption of Intranasal Salmon Calcitonin (SCT) in Healthy Subjects

In young healthy subjects salmon calcitonin (SCT), intranasally administered, increased in serum as a function of the drug administration time. The serum concentration of a 400 IU SCT dose monitored 10 min after dosing was statistically more significant when inhaled at 0000 than at other, more conventional, administration times (morning or evening). Following dosing at certain times during the day, the serum SCT was less or even questionable with the dose and under the study conditions selected. Dosing without consideration of timing may lead to reduced effect or lack of effect or perhaps ambiguity or controversy regarding the possible circumstance of a “non-absorbent subject”. The circadian frequency appears to be a critical determinant of intranasal SCT absorption suggesting administration time to be an important factor in the cost/benefit ratio without the unpleasant side effects sometimes experienced through parenteral routes.     

Some factors influencing the sensitivity of body temperature to activity in neonates

In adult humans, core temperature is influenced by activity; the sensitivity of core temperature to such effects shows a phase dependence and is also influenced by the environment and whether the individual is asleep or awake. We have investigated if similar effects are evident in neonates, in whom thermoregulation and the circadian rhythm of core temperature are not fully developed. Eleven full-term, healthy babies were studied singly (light 07:00-19:00) at 2 days of age and again 4 weeks after birth; between these times, they were tended routinely on a communal ward. On study days, 10-minute recordings were made of rectal and skin (abdominal) temperature, heart rate (HR), and behavioral state. Sensitivities of the temperatures to activity (“arousal”) were assessed throughout the 24h by measuring the gradient of (temperature/HR). Sensitivities measured at 01:00, 05:00, 09:00, 13:00,17:00, and 21:00 were used as dependent variables in stepwise regression and linear regression analyses, with “subjects” “light versus dark”, “behavioral state”, and “difference between time of measurement and the acrophase of the endogenous component of the temperature rhythm” (ignoring sign) as possible predictors. (Acrophases of the temperature rhythms had been estimated from 24h data purified using the behavioral state record.) Light versus dark acted as a significant predictor of the sensitivity of rectal temperature to arousal on day 2 and week 4, the sensitivity increasing in the light, and there was limited evidence for behavioral state acting as a predictor on day 2. Neither factor was a significant predictor when the sensitivity of the babies' skin temperatures to arousal was investigated. There was also some evidence that the difference between the time of measurement and the temperature acrophase acted as a predictor of sensitivity to arousal in both rectal (day 2) and skin (week 4) temperature, with larger differences decreasing the sensitivity. These results indicate that there are masking effects on body temperature due to arousal in neonates, the size of which depends on both internal and external factors. However, this sensitivity of temperature to arousal shows differences from the sensitivity of temperature to physical activity in both adult humans and adult mice. One possible explanation of this result is that temperature regulation and the circadian system are not fully developed in humans at this age. (Chronobiology International, 17(5), 679-692, 2000)     

Enclosure design and space utilization by Indian leopards (Panthera pardus) in four zoos in Southern India

Enclosure design and the use of enclosure space influence the activity budget of captive leopards. The study laid out in grids all enclosures on the base map and segregated these grids into 4 zones. Every 5 min, the study recorded the proportion of time spent in these zones with the leopards' behavior. Captive leopards most frequently used the “edge ”zone. Almost all leopards used the edge zone for stereotypic pacing, the “back ”zone for resting, and the “other ”zone for activity. The study positively correlated the proportion of time spent in the “enriched ”zone with activity levels exhibited by leopards housed in some enclosures and with resting in others. Thus, the study segregated structural objects in the enriched zone into activity-related features (e.g., logs) and rest-related features (e.g., trees and sleeping platforms). Compared with individuals housed in barren enclosures, leopards housed in structurally enriched on-exhibit enclosures exhibited higher levels of activity. Enclosure design was found to be an important factor influencing the welfare of leopards in captivity.