EXTRACTION,ASSAY AND SOME PROPERTIES OF FLORIDOSIDE PHOSPHATE SYNTHASE FROM PORPHYRA PERFORATA (RHODOPHYTA)1

A suitable method for extraction of floridoside phosphate synthase (FPS, UDP-galactose: sn-3-glycerol phosphate: 1→2′α-D-galactosyl transferase)from Porphyra perforata J. Ag. was developed. Two assay methods for enzyme activity were utilized, one measuring the amount of floridoside formed by using gas-liquid chromatography, the other measuring the sn-3-glycerol phosphate-dependent formation of UDP; both assays gave similar results. FPS is a soluble protein, and FPS activity in the extract as determined by the amount of product formed in vitro compared well with the in vivo rate of floridoside synthesis (4–7 μMmol product formed·h^?1·g^?1 fresh wt). The rate of product formation in vitro was linear up to 45 min and proportional to protein concentration in the assay mixture. The temperature optimum was 30–35° C. FPS was active over a range of pH values from 7.0–8.5. It was stable in concentrated solutions in the presence of 0.3 M ammonium sulfate, but activity was lost in diluted solution (protein concentration below 0.2 mg·mL^?1) or below 0.2 M ion strength. The data suggest that FPS may be an oligomeric protein which occurs free in the cytoplasm or loosely bound to a membrane. It may also be a regulatory protein controlling the overall rate of synthesis of floridoside in vivo.     

FLORIDOSIDE AS A CARBON PRECURSOR FOR THE SYNTHESIS OF CELL‐WALL POLYSACCHARIDE IN THE RED MICROALGA PORPHYRIDIUM SP. (RHODOPHYTA)1

Although red algae are known to be obligatory photoautotrophs, the red microalga Porphyridium sp. was shown to assimilate and metabolize floridoside. A pulse‐chase experiment with [¹⁴C]floridoside showed that at the end of a 240‐min pulse, 70% of total ¹⁴C‐uptake by the cells remained in the floridoside fraction. To evaluate the assimilation of floridoside by Porphyridium sp. cells, we exposed Porphyridium sp. not only to [¹⁴C]floridoside but also to its constituents, [¹⁴C]glycerol and [¹⁴C]galactose, as compared with [¹⁴C]bicarbonate. The extent of incorporation of [¹⁴C] galactose by the Porphyridium sp. cells was insignificant (50–80 dpm·mL^?1), whereas uptake of ¹⁴C from [¹⁴C]glycerol into the algal cells was evident (2.4 × 10³ dpm·mL^?1) after 60 min of the pulse. The pattern of ¹⁴C distribution among the major constituent sugars, xylose, glucose and galactose, of the labeled soluble polysaccharide was dependent on the ¹⁴C source. The relative content of [¹⁴C]galactose in the soluble polysaccharide was highest (28.8%) for [¹⁴C]floridoside‐labeled culture and lowest (19.8%) for the [¹⁴C]glycerol‐labeled culture. Upon incubation of [¹⁴C]floridoside with a crude extract of a cell‐free system prepared from nonlabeled cells of Porphyridium sp., the label was indeed found to be incorporated into the sulfated polysaccharide. Our results suggested that the carbon metabolic pathway in Porphyridium sp. passes through the low molecular weight photoassimilatory product—floridoside—toward sulfated cell‐wall polysaccharide production.     

Diurnal regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase activity and its content in Norway spruce needles

Diurnal changes of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activity and its content were measured to find the mechanism of RuBPCO activity regulation in Norway spruce needles. Both initial and total RuBPCO activities as well as the activation state had a typical pattern with two peaks in the morning and afternoon, respectively, and a midday depression. On the 19 October, RuBPCO content decreased during the day from 3.1 to 1.4 g m⁻², while on the 20 October it was approximately constant both in the morning and in the afternoon (2.7 g m⁻²). Neither initial nor total activity of RuBPCO copied irradiances. Relatively low morning and evening values of total activities indicate that nocturnal inhibitor CA1P is important in Norway spruce. However, the midday depression of total activity indicates that besides CA1P there function some other inhibitors of RuBPCO. In addition, the diminution of RuBPCO content during the day may indicate repression of its gene expression.     

Behavioral rhythms of the Japanese newts, Cynops pyrrhogaster, under a semi-natural condition

 Locomotor activity rhythms of the Japanese newt, Cynops pyrrhogaster, were recorded under a semi-natural condition using phototransistor systems. The daily activity rhythm showed a seasonal change: the locomotor activity was mainly diurnal (active during the daytime) from spring to early summer; mainly nocturnal (active during the night-time) from summer to autumn; and showed either a diurnal or nocturnal pattern, depending on the ambient temperature, in winter. To analyze the daily activity in detail, we observed the behavior of a group of newts (three males, three females) throughout 24 h. Four types of behavior (respiration, feeding, mating, and resting on the land) were observed. Each behavior had daily rhythms and showed a seasonal change. The behavior on land showed mainly a nocturnal or bimodal pattern (activity rhythms with two peaks) throughout the year and was more frequently observed in summer. Mating behavior also showed a seasonal change: high activity in spring, with peaks in the early morning and evening, but no activity in summer. Except in winter, feeding and respiratory behavior showed no seasonal changes in either activity period or frequency. Coupling between behavior and the clock seems to be weak in the Japanese newt because of indistinct daily rhythms and frequent phase changes of locomotor activity in water. Physical factors such as humidity and temperature seem to affect strongly the daily activity of the newts. Received: 21 April 1997 / Accepted: 1 September 1997     

NUTRIENT UPTAKE KINETICS IN PHYTOPLANKTON: A BASIS FOR NICHE SEPARATION1

Daily patterns of incorporation of carbon dioxide and inorganic phosphate were measured in phytoplankton from Lake George, New York, in January and February 1972. Rates of photosynthesis oscillated in phase for the entire assemblage and for individual cells of the diatoms Asterionella, Tabellaria, and Fragilaria. The photosynthetic capacity was maximal in the early afternoon. Daily patterns of phosphate uptake were also rhythmic. At ambient concentrations the assemblage takes up phosphate maximally in the morning while individual cells of the large diatom take it up maximally in the evening. A kinetic analysis of phosphate uptake indicated 2 velocities of uptake for cells of Tabellaria and Fragilaria: a hyperbolic function at small and an apparently linear relationship at relatively large concentrations. The large diatoms, in contrast to the total assemblage, functioned maximally at 2 separate times of the day: in the evening at ambient levels and in the morning at 0.4 μM and larger concentrations. Temporal stratification of the nutrient niche may be achieved by several uptake mechanisms in the algal cell that function at different times of the day or with a variable uptake velocity.     

Circadian rhythm in Schistosoma haematobium egg excretion

The usual peak daily egg excretion period in persons harbouring Schistosoma haematobium is the late morning and early afternoon. No evidence was found to support a hypothesis that the rhythm is mechanical in origin being due to rapid increase of bladder activity in the early morning.A partial reversion of the rhythm occurred when day shift workers changed to night shift.The possible role in the circadian rhythm of host factors—temperature and metabolism and of proteolytic enzyme secretion by miracidia are discussed.     

Day length and evening temperature predict circannual variation in activity duration of the colony of the Indian cliff swallow,Hirundo fluvicola

In avian species, circannual rhythms, in a number of biological variables, including locomotor activity, have been studied under both laboratory and natural environmental conditions. However, information on circannual rhythm in daily activity duration of Indian cliff swallow (Hirundo fluvicola) is not available. It is a communal mud nesting, non-migratory species and inhabits low under arch bridges. Although it figures in the IUCN Red List under the least concerned category, it is important to study its behavioral ecology that may be of utmost importance from conservation stand point. In the present investigation, we examined the circannual rhythm in daily activity duration in this species at a communal roosting site under Kharoon river bridge on the Raipur–Bhilai highway (21°15′07.54′′N; 81°32′30.65′′E), Raipur, India, for eight consecutive days, every month from August 2009 to October 2011. On each study day, observations were made in two sessions, morning and evening. We monitored and recorded emergence time of the first bird from the nest in the morning and entry time of the last bird to the nest in the evening. We also recorded the ambient temperature and humidity of the study site simultaneously. Average daily activity duration (the time difference between the first bird’s emergence time and the last bird’s entry time from/to the nest, respectively) was computed for each month. A significant circannual rhythm in activity duration was validated. The peak of activity rhythm occurred on 22nd June with a spread between 19th June and 25th June. The activity duration was the shortest in winter and the longest in summer. The daily activity duration was positively correlated to the day length, sunset time, and morning and evening temperatures; whereas negatively correlated with the sunrise time, and morning and evening humidity. The multiple linear regression models suggest that day length alone explains 96% of the total variance in activity duration, whereas only 0.5% variance was attributable to evening temperature. We conclude that the day length is the strongest predictor of circannual variation in daily activity duration of the colony of Indian cliff swallow; whereas the evening temperature has very little effect. Further, we recommend that comparable studies under natural conditions might be very helpful to explore the effect of environmental cues on other intriguing behavioral decisions made by this and other avian species.     

A QUANTITATIVE STUDY OF THE DAILY ACTIVITY OF THE WARTHOG IN THE QUEEN ELIZABETH NATIONAL PARK, UGANDA

Warthog activity was recorded at 4-min intervals for continuous periods of 14 h on three consecutive days. The major activities were feeding or lying down and walking. The amount of time spent in feeding is related to the size of the animal and maximum-intensity feeding occurs in the early morning and in the late afternoon and evening. The animals drank usually only once each day. Adult females urinate much more often than males and this is thought to be a behavioural adaptation whereby the males can tell whether or not a female is in heat.     

Evidence of daily spawning in natural populations of the New Zealand snapper Pagrus auratus (Sparidae)

Synopsis New Zealand snapper, Pagrus auratus, were captured by trawling from NE New Zealand over two successive spawning seasons, and examined for acute temporal changes in gonad condition. Fish with oocytes completing final oocyte maturation predominated during the morning, with a peak in ovulated fish occurring just after midday. Afternoon catches were dominated by fish in which the most advanced oocytes had yet to begin final maturation. This suggests that ovulation is synchronised to occur soon after midday, and the high proportion (up to 100% of the catch) of fish with particular gonad stages captured at any one time indicates that daily spawning involves most of the population. Diurnal changes in oocyte diameter support a daily spawning rhythm, with numbers of large hydrated oocytes peaking in the late morning, followed by the disappearance of these oocyte stages in the afternoon. Snapper captured alive by longlining were returned to the laboratory to examine the relationship between ovulation and probable time of spawning. Unovulated fish generally ovulated close to midday on the day of capture (morning captures), or the following day (afternoon captures). The viability of ovulated eggs (proportion undergoing division after fertilization) decreased markedly after oviduct residence times of over 8 hours. This suggests that natural spawning occurs before the late evening. The results of this study are consistent with anecdotal evidence suggesting that spawning occurs every day during the late afternoon or early evening, and is similar to the reproductive patterns displayed by a number of closely related sparids. Department of Zoology, University of Otago, PO Box 56, Dunedin, New Zealand     

SALINITY TOLERANCE,BILIPROTEINS, AND FLORIDOSIDE CONTENT OF COMPSOPOGON COERULEUS (RHODOPHYTA)1

The freshwater red alga Compsopogon coeruleus (Balbis) Montague is capable of growing and reproducing in salinities up to 35 ppt. Increased accumulation of floridoside (D-galactopyranosyl glycerol) parallels increase in salinity. Compsopogon phycobilisomes contain an unusual B-phycoerythrin completely lacking in phycourobilin chromophores, but with a pigmented γ subunit. The α, β, and γ subunits of this phycoerythrin all carry phycoerythrobilins. These results suggest that C. coeruleus is secondarily adapted to freshwater from marine habitats.     

The effect of training at a specific time of day: a review

This article focuses on physical performances after training at a specific time of day. To date, although the effect of time of day on aerobic performances appears to be equivocal, during anaerobic exercises, the effect of time of day has been well established with early morning nadirs and peak performances in the late afternoon. These diurnal rhythms can be influenced by several factors such as the regular training at a specific time of day. Indeed, regular training in the morning hours may increase the lower morning performances to the same or even higher level as their normal diurnal peak typically observed in the late afternoon by a greater increase of performance in the evening. However, regular training in the evening hours may increase the morning-evening (i.e., amplitude of the rhythm) difference by a greater increase of performance in the late afternoon. Therefore, adaptations to training are greater at the time of day at which training is regularly performed than at other times. Nevertheless, although modifications in resting hormones concentrations could explain this time-of-day specific adaptations, precise information on the underlying mechanisms is lacking.     

Low MW carbohydrates and ions in rhodophyceae: Quantitative measurement of floridoside and digeneaside

The content of the heterosides floridoside and digeneaside and of the main ions Na⁺, K⁺, and Cl^? was estimated in 20 species of the Rhodophyceae. Methods for quantitative determination of the heterosides are described. The floridoside content is in the range of 1.5–8% on a dry weight basis (Catenella: up to 22%); the content of digeneaside, exclusively found in species of the Ceramiales, is lower, in the range of 1–2.2% on a dry weight basis. All species investigated have Cl^? as main anion, while there is a remarkable diversity in cation composition. Na⁺ was the major cation in 12 of the species investigated, the others having K⁺ as main cation.     

EFFECT OF NUTRIENT DEPRIVATION AND RESUPPLY ON METABOLITES AND ENZYMES RELATED TO CARBON ALLOCATION IN GRACILARIA TENUISTIPITATA (RHODOPHYTA)1

The starch content of red algae normally increases during nitrogen limitation. Based on this we hypothesized that nutrient deprivation would result in an increased activity of starch‐synthesizing enzymes and a decrease in the activity of starch‐degrading enzymes, with the opposite scenario when nutrients were sufficient. We therefore examined the effect of the nutrient status of Gracilaria tenuistipitata Chang et Xia on the content of starch and floridoside and on the activity of enzymes involved in the allocation of carbon into starch, floridoside, and agar; floridoside phosphate synthase and α‐galactosidase involved in synthesis and degradation of floridoside; starch synthase and starch phosphorylase involved in the metabolism of starch; uridine 5′‐diphosphate (UDP)‐glucose pyrophosphorylase; adenosine 5′‐diphosphate‐glucose pyrophosphorylase; UDP‐glucose 4‐epimerase; and phosphoglucomutase. During the period of nutrient limitation the starch and floridoside content increased, as did dry weight and C/N ratio, whereas growth rate and protein content decreased. A general decrease in the enzyme activities during nutrient limitation was also observed, indicating a decrease in overall cellular metabolism. The addition of nutrients caused an increase in enzyme activities and a decrease in the contents of starch and floridoside. Of the enzymes examined, only the activity of UDP‐glucose pyrophosphorylase increased during nutrient limitation and decreased abruptly after nutrient addition. This implies a regulatory role for this enzyme in the supply of UDP‐glucose for starch synthesis. It also supports our suggestion that UDP‐glucose is the substrate for starch synthesis in red algae. This assertion is further strengthened by the observation that of the potential starch synthases only the UDP‐glucose starch synthase could support the observed rate of starch synthesis.     

Collegiate rowing crew performance varies by morningness-eveningness

During adolescence and early adulthood, most humans are predisposed developmentally, both biologically and socially, toward evening/night activity. The morningness-eveningness (M-E) tendency to be an evening-preferring (E-type) rather than a morning-preferring (M-type) or intermediate/neither (N-type) "chronotype" may affect athletic performance at various times of day. This study evaluated M-E effects on rowing performance of an intact, experienced, university club crew with near-daily early morning (0500-0700 hours) and late afternoon (1630-1800 hours) training schedules. The hypothesis tested was that chronotype would modify circadian effects during morning and afternoon performances. Eight men and eight women (mean age 19.6 +/- 1.5 years) were tested in a randomized, counterbalanced design. A standard qualifying 2000-m ergometer rowing sprint and a nonroutine standing broad jump task were measured during early morning and late afternoon, separated by 3 days of rest. Each subject's chronotype was determined using two standard self-rating M-E scales, resulting in eight E-type (three women/five men), four M-type (two women/two men), and four N-type (three women/one man) subjects. The rowing results show that E-type and N-type subjects did not differ between morning and afternoon rowing performances, whereas M-type subjects rowed significantly faster in the morning. In contrast, the standing broad jump showed no consistent time-of-day or chronotype effect. These findings suggest that basic performance timing in young athletes is determined to some extent by naturally occurring M-E predispositions. Further, modification of time-of-day influences may be possible by routine practice at the same time each day, as was suggested here by the absence of evening superiority in performances. Understanding their personal M-E tendencies could allow young athletes to arrange training schedules at specific times of day to help counteract any natural circadian influences that might work against their performance.     

Windows of opportunity and the temporal structuring of foraging activity in a desert solitary bee

1. Females of the desert solitary bee Anthophora pauperata collect nectar and pollen almost exclusively from Alkanna orientalis (Boraginaceae). The bee and plant are found together in the early spring, living in the bottom of steep-sided wadis (dry river valleys) at an altitude of 1500 m in Egyptian Sinai. 2. Female A. pauperata showed clear morning and afternoon peaks in foraging activity, separated by a 2–3 h midday period spent in their underground nests. This study analyses the following in order to identify the factors structuring this daily pattern: thermal aspects of the bee and its environment, temporal patterns of resource provision by the plant, and female nectar and pollen foraging behaviour. 3. Although A. pauperata can generate substantial heat endothermically, morning and evening ambient temperatures well below 10 °C defined a thermal window within which foraging occurred. Maximum air temperatures were moderate (25–30 °C), and examination of the physiology and behaviour of A. pauperata suggests that the midday reduction in flight activity was not due to thermal constraints. 4. Alkanna orientalis produces protandrous hermaphroditic flowers. Female A. pauperata collected pollen from male-phase flowers and harvested nectar preferentially from female-phase flowers. Although the nectar standing crop was relatively constant throughout the day, pollen availability peaked strongly in the early afternoon. 5. Female A. pauperata visited young male-phase flowers as soon as they opened, generating an early afternoon peak in pollen foraging activity and depleting the pollen standing crop rapidly. A morning peak in pollen foraging occurred when females gleaned remnant pollen from flowers that had opened the previous day. Pollen availability in the morning was far lower than in the early afternoon, and the time taken to collect a full pollen load in the morning was significantly longer. Collection of pollen in the morning despite very low resource availability suggests that pollen may be a limiting resource for A. pauperata. 6. In contrast to many existing examples of bimodal activity patterns in highly endothermic bees, the bimodal activity patterns of female A. pauperata appear to be driven not by thermal considerations but by daily patterns of pollen release from its principal food source.     

The dual-oscillator system of Drosophila melanogaster under natural-like temperature cycles

Dual-oscillator systems that control morning and evening activities can be found in a wide range of animals. The two coupled oscillators track dawn and dusk and flexibly adapt their phase relationship to seasonal changes. This is also true for the fruit fly Drosophila melanogaster that serves as model organism to understand the molecular and anatomical bases of the dual-oscillator system. In the present study, the authors investigated which temperature parameters are crucial for timing morning and evening activity peaks by applying natural-like temperature cycles with different daylengths. The authors found that the morning peak synchronizes to the temperature increase in the morning and the evening peak to the temperature decrease in the afternoon. The two peaks did not occur at fixed absolute temperatures, but responded flexibly to daylength and overall temperature level. Especially, the phase of the evening peak clearly depended on the absolute temperature level: it was delayed at high temperatures, whereas the phase of the M peak was less influenced. This suggests that the two oscillators have different temperature sensitivities. The bimodal activity rhythm was absent in the circadian clock mutants Clk(Jrk) and cyc(01) and reduced in per(01) and tim(01) mutants. Whereas the activity of Clk(Jrk) mutants just followed the temperature cycles, that of per(01) and tim(01) mutants did not, suggesting that these mutants are not completely clockless. This study revealed new characteristics of the dual-oscillator system in Drosophila that were not detected under different photoperiods.     

Morningness-eveningness scores predict outcomes differentially for depressed patients attending morning vs. afternoon day treatment streams

ABSTRACT

At the Center for Addiction and Mental Health (CAMH) Integrated Day Treatment (IDT) program, each patient attends either a morning stream or an afternoon stream, but not both. We examined whether subjective chronotype, or the time of day an individual prefers to be most active and alert, predicted treatment outcomes differentially in depressed patients attending the morning vs. afternoon IDT streams. The Horne-Östberg Morningness-Eveningness Questionnaire (MEQ) was administered before IDT treatment to 203 consecutive patients experiencing a major depressive episode. Multiple regression was used to predict change in depression and quality of life scores based on treatment stream (morning or afternoon), baseline MEQ scores and the treatment stream by MEQ interaction. The treatment stream by MEQ interaction was a highly significant predictor of both depression and quality of life change scores. Post-hoc analyses based on established MEQ categories revealed that definite evening chronotypes had significantly better responses in the morning stream than did morning chronotypes, and significantly worse responses in the afternoon stream relative to moderate evening or neutral chronotypes. There were insufficient morning chronotypes in the afternoon stream to assess clinical responses for this subgroup. In the morning stream only, there was a significant positive correlation between the change in MEQ scores after four weeks of IDT treatment (i.e. a shift to greater morningness) and the decrease in depression scores (r = .36, p = .003), consistent with a therapeutic phase advance in circadian rhythms. In sum, these preliminary data suggest that definite evening chronotypes may have the greatest relative benefit from attending the morning vs. afternoon IDT stream. As patients currently select which IDT stream they will attend, future work based on randomized treatment assignment and using passive actigraphy to assess circadian phase is currently planned to extend these preliminary findings.     

Diurnal activity rhythms of the subterranean termiteAnacanthotermes vagans (Hagen) under laboratory and field conditions of the Kuwait desert

Daily foraging activity ofAnacanthotermes vagans was tested using toilet paper rolls as bait. The highest activity was recorded at midnight and during the hours of the early morning in both summer and winter seasons. In spring the time of highest activity was reversed to take place at midday, afternoon and early evening. Aktograph records in the laboratory demonstrated that individual workers are active most of the day and not naturally entrianed by photoperiods. Daily activity of groups of 10 workers was not statistically different from that of individual workers.     

Daily and seasonal rhythms of locomotor activity and adrenal function in male hedgehogs(Erinaceus europaeus L.)

Abstract

Male hedgehogs were maintained in outdoor parks, under natural climatic conditions. The general locomotor activity was recorded over one year in five hedgehogs, using an infra‐red producer‐receptor system registering entering or leaving the nest box in relation to time. Simultaneously, thecortico‐adrenal activity was studied monthly by sampling without disturbing, at 4 h intervals over a period of 24 h, in groups of 6 animals with a catheter inserted in the left carotid artery. The corticosteroids were assayed using a competitive protein binding technique.

In our study, hedgehogs living in Western Central France showed:

(1) from February‐March to August‐September a daily nocturnal locomotor activity well synchronized with sunset and sunrise,

(2) between August‐September and February‐March an alternation of resting and activity periods which constitute the hibernating period. The activity periods occur still during the night, but there are no relationships with sunset or sunrise,

(3) from February to October, a nycthemeral rhythm of plasma corticosteroids wi th an evening maximum and a morning minimum, as in laboratory animals with nocturnal locomotor activity,

(4) after October, during hibernation, significant variations of plasma corticos teroids with an irregular.pattern (shifting of the maximum or biphasic profile),

(5) a clearly annual cycle of the daily plasma corticosteroids levels, with a considerable increase in autumn (maximum in December), a significant decrease in January and a minimum from January to May.

These daily and seasonal variations of locomotor activity and cortico‐adrenal activity are related to the seasonal variations of the environment.     

Daily Changes in the Competence for Photo- and Gravitropic Response by Potato Plantlets

Competence for phototropic (PT) and gravitropic (GT) bending by potato plantlets grown in vitro manifests regular daily changes indicating possible involvement of circadian regulation. Unilateral stimulation of plantlets with blue light at dawn resulted in moderate PT response regarding both attained curvature and long lag phase. The PT response was the strongest between 8:00 and 12:00 h. Throughout the afternoon and in the evening, bending rate and maximal PT curvature declined significantly until 23:00 h. The GT response was fastest and strongest for plantlets stimulated early in the morning and late in the evening. During the rest of the day, GT competence did not change much apart from a minimum at 15:00. In conditions comprising either prolonged day or prolonged night, plantlets appeared to maintain rhythmicity of competence for PT and GT at least in the short-term. Introduction of a dark period prior to the tropic stimulation at 11:00 h when both PT and GT responses were strong resulted in the opposite effect: PT was depressed, and GT was enhanced. There was a time threshold of 60 min for the duration of the dark period so the plants can sense interruption in the daylight. Levels of relative expression of a PHOT2 gene indicate rhythmic daily changes. The PHOT2 gene was present at high levels during morning hours and late in the evening. As the mid-day and the afternoon hours approached, PHOT2 expression decreased and reached a daily minimum at 17:00 h. We believe that our data offer strong support for the conclusion that there is an involvement of circadian rhythms in control of both PT and GT.