Ultrastructural circadian variation of mast cells in the pinna of the mouse

It has been found previously under the light microscope that there was a circadian variation in mast cell number in the pinna of mice. The mast cell number was increased at 18.00 h and decreased at 06.00 h. In the current study, 5 mice of each group were synchronized for 4 weeks with a standard lighting regimen (light: 06.00-20.00 h; dark: 20.00-06.00 h). Both pinnas of every mouse of each group were removed at 06.00, 12.00, 18.00 and 24.00 h, respectively. Under the electron microscope, it was observed that more degranulated mast cells were found at 06.00 h and more intact mast cells were visible at 18.00 h. It appeared that the mast cell numbers, decreased and increased under the light microscope, were due to mast cell degradation and regranulation. This finding suggests that a functional circadian variation exists in the mast cell under physiological conditions.     

Circadian rhythms of folate and vitamin B12 concentration in relation to convulsive thresholds of mice

Circadian rhythms of folate and vitamin B₁₂ concentrations in the liver, brain and blood of Swiss mice have been determined. The relation of the changes in vitamin concentration to circadian rhythms in locomotor activity, drinking activity and convulsive thresholds (maximal electroshock seizure threshold) have been determined also. Both free folic acid (FFA) and total folic acid (TFA) of liver and blood showed minimum values at 21.00–24.00 h and maximum values at 06.00–09.00 h. Liver TFA declined at a steady rate from the peak value at 09.00 h whereas liver FFA and plasma folate maintained constant values from 09.00–18.00 h then declined rapidly. Brain folate (TFA and FFA) showed no rhythm. The concentrations of vitamin B₁₂ in plasma, liver and brain showed only minor fluctuations. Locomotor and drinking activities showed very similar rhythms with a sustained period of high activity between 12.00–21.00 h followed by a much shorter second period of high activity (23.00–03.00 h). Convulsive threshold declined during the first period of increased locomotor and drinking activities reaching a minimum value at 21.00 h thus just preceding the nadir in folate concentrations in liver and blood.     

Circadian rhythms in the incidence of apoptotic cells and number of clonogenic cells in intestinal crypts after radiation using normal and reversed light conditions

Variations in the number of radiation-induced morphologically dead or dying cells (apoptotic cells) in the crypts in the small intestine of the mouse have been studied throughout a 24-h period under a normal light regimen (light on, 07.00-19.00 h; light off, 19.00-07.00 h). A clear circadian rhythm was displayed in the apoptotic incidence 3 or 6 h after irradiation for each gamma-ray dose studied (range 0.14-9.0 Gy). The most prominent circadian rhythm was obtained after 0.5 Gy. The peak time of day for inducing apoptosis was 06.00-09.00 h, and the trough occurred at 18.00-21.00 h. Some mice were also transferred to a room with the light cycle reversed, and were irradiated on different days after the transfer. The apoptosis induced by 0.5 Gy or 9.0 Gy, or the number of surviving crypts (microcolonies) after 11.0 Gy or 13.0 Gy was examined. The transition point for reversal (i.e. the switch time from the normal-light pattern to the reversed-light pattern) of the circadian rhythm in apoptosis (after 0.5 Gy) occurred 7 days after the transfer and the rhythm was reversed by 14 days. The rhythm for crypt survival (i.e. for clonogenic cell radiosensitivity) was disturbed on 1 day and the transition point for reversal occurred 3 days after the transfer. The rhythm became reversed by 7 days. These observations are discussed in relation to the identity of clonogenic cells, (functional) stem cells, proliferating transit cells and the cells sensitive to small doses of radiation (i.e. hypersensitive cells) in the crypt.     

The light cycle as a modulator of aminopyrine demethylation by liver from normal,orchiectomized or adrenalectomized rats

Abstract

As part of studies on the interactive effects of light and endocrine factors on hepatic mixed‐function oxidases, the aminopyrine demethylase activity of liver microsomes and the serum concentrations of corticosterone and testosterone were measured in adult male rats (sham‐operated, castrated or adrenalectomized) exposed for 21 days to one of the following types of environmental lighting: (1) Normal light cyle (L from 09.00 to 21.00 h); (2) Reverse light cycle; (3) Constant light (LL) or (4) Constant darkness (DD). One half of the animals in each of the 12 groups was killed at 06.00 h and the other at 18.00 h on the last day of treatment. A 3‐way analysis of variance, multiple comparisons and correlations allowed the following conclusions: (1) The type of lighting had the most significant effect on demethylase activity, its highest values corresponding to periods of darkness (particularly in the rats exposed to a reverse light cycle) and its lowest to LL. (2) Castrated rats as a group had the lowest demethylase activity. (3) Time of death was a significant factor, particularly for the rats in a reverse light cycle (higher activity at 18.00 than at 06.00 h). (4) There was a significant and positive correlation between aminopyrine demethylase and the rates of 7α‐hydroxylation of 3β‐hydroxy‐5‐androsten‐17‐one previously reported for the same animals. (5) There was no significant correlation between demethylase activity and serum corticosterone or testosterone.     

Compartmentalized mast cell degranulations in the ovarian hilum, fat pad, bursa and blood vessel regions of the cyclic hamster: relationships to ovarian histamine and blood flow.

Ovaries from hamsters on each day of the oestrous cycle at 09.00 h were observed for the number of mast cells, the pattern of mast cell degranulation, histamine concentration and blood flow. On day 4 (pro-oestrus), ovaries were also observed at 9.00, 15.00 and 21.00 h. Mast cell degranulation was evaluated by 3 criteria: (1) no degranulation = less than 5 granules dispersed from the cell; (2) moderate degranulation = 5 or more granules dispersed but less than 15, and (3) extensive degranulation = 15 or more granules released. Blood flow was determined using radio-active microspheres in anaesthetized animals. Mast cells were observed in fat pad (beyond 2 mm of the bursal mesothelium), bursa (within 2 mm of the bursal mesothelium), hilum and near ovarian blood vessels (these 4 regions are collectively called the ovarian complex). The distribution of ovarian mast cells was not uniform. Most mast cells were near ovarian blood vessels (42.2%) and in the fat pad (37.2%). A moderate number of cells were in the bursal wall (20%) and only a few cells were observed in the hilum (0.64%). Mast cell number remained unchanged on days 1-4 of the cycle in each ovarian compartment. However, summation of the number of mast cells in the entire ovarian complex revealed a significant decline in number at 15.00 h on pro-oestrus. Alterations in mast cell degranulation were primarily restricted to 2 periods of the cycle (pro-oestrus and di-oestrus). An increase in moderate but not extensive degranulation was observed in only the fat pad and bursa on day 2 when compared with day 1 values. In most ovarian compartments on pro-oestrus, degranulation was higher than on any other day of the cycle. At 15.00 h on pro-oestrus, extensive degranulation in bursa, fat pad and blood vessel regions (but not hilum) coincided with an increase in ovarian histamine and decline in number of mast cells; ovarian blood flow also increased at the time but remained unchanged the remainder of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Circadian rhythm of aldosterone in dairy cattle during the summer

Twelve Holstein heifers, pregnant from 120–150 days were used to study the circadian rhythm of aldosterone, cortisol, progesterone, sodium and potassium in dairy cattle during the summer in Louisiana. Cortisol was not significantly influenced by time (time 1 = 06.00 h). Aldosterone, sodium, potassium and progesterone changed significantly (P<.01) with time. Aldosterone peaked (116.5±17.2 pg/ml) at 08.00 h and then generally declined to 16.00 h (26.7±2.0 pg/ml). Sodium generally increased from 06.00 h (320.1±7.3 mg%) to 18.00 h (377.9±6.1 mg%), and then declined. Potassium generally increased from 06.00 h (20.9±0.5 mg%) to 22.00 h (23.0±0.3 mg%). Progesterone generally increased from 07.00 h (2.8±0.4 mg/ml) to 24.00 h (7.5±1.4 mg/ml). Aldosterone was significantly related to temperature associated with the time of the day samples were taken (r = 0.66, P<.02).     

Some observations on the diurnal variation of mitosis in the stratified squamous epithelium of wounded tympanic membrane

Summary A study of mitosis in the stratified squamous epithelium of the tympanic membrane of the guinea-pig was made after wounding this organ at 09.00 h in one series of animals and at 21.00 h in a second series. It was found that the diurnal variation of mitosis was abolished by the injury. This finding corresponds with a basic requirement of the chalone-adrenaline hypothesis which is considered to be a factor in the mitotic control of epidermal cells. It was also found that the maximum number of dividing cells was at the wound edge in the group wounded at 09.00 h, which also agrees with the chalone concept. In the group wounded at 21.00 h, however, the initial mitotic response was in cells distally placed from the wound edge which is more difficult to explain. Perhaps these cells are inhibited mitotically by the factors normally producing a low nocturnal mitotic rate and undergo migration to cover the defect in the first instance, and only divide at a later time when the mitotic inhibition is lifted. It appears that the factors responsible for the increased mitotic rate after wounding are different from those responsible for the diurnal variation.The author wishes to express his thanks to Professor R.M.H. McMinn for his support of this work     

Circadian variations of the plasma proteins in the adult male rat under natural synchronisation (author's transl)]

Circadian rhythms of blood total proteins, albumin, alpha 1-, alpha 2-, beta- and gamma-globulins were documented in 80 Wistar SPF adult male rats, synchronized by natural light (06.00-18.00 h) and darkness (18.00-06.00 h) during the month of October 78. Blood was sampled at four fixed time points in the 24 h scale (i.e. 04.00, 10.00, 16.00 or 24.00 h). Total proteins, albumin, alpha 2- and gamma-globulins showed a statistically significant rhythm with a maximum at 04.00 h. The data obtained in anaesthetized rats under artificial synchronization and in man are compared, taking into account that the rat is a nocturnally active rodent. The present work partially confirms the hypotheses of previous chronopharmacological studies, thus e.g. curarizing substances have a mimimum activity when the protein plasma level, especially albumin, is the highest.     

A morphometric study of 24-hour variations in subcellular structures of the rat pancreatic acinar cell

Summary Subcellular structures of pancreatic acinar cells were examined at six evenly spaced time points in the 24-h period (light cycle: 06.00 h–18.00 h) in four Wistar male rats at each time point. At each sampling point, the area and circumference of acinar cell bodies and the area, number and circumference of their cytoplasmic organelles were measured using a semiautomatic computer system for morphometry and a point-counting method.The area, number and circumference-area ratio of the cytoplasmic organelles were subject to strong circadian variations, and the cellular area and circumference exhibited weak circadian variations. Variation pattern of the cytoplasmic organelles suggested an intracellular route for secretory proteins during a 24-h span. From the results it was possible to divide the 24-h period into three stages. 1. The resting or protein synthetic stage (00.00 h to 08.00h): the area of the rough surfaced endoplasmic reticulum (rER) was strongly increased, and that of zymogen granules was clearly decreased. 2. The granule accumulation stage (08.00h to 16.00h): the area of the rER was markedly decreased; that of zymogen granules was increased. 3. The secretion stage (16.00 h to 00.00): as a result of the release of zymogen granules from the acinar cell, the area of zymogen granules decreased, and that of the rER increased. The relationship between the area of the rER and zymogen granules varied in a reciprocal manner. Other cytoplasmic organelles, namely the Golgi complex, condensing vacuoles, mitochondria and lysosomes also varied prominently during the 24-h span, corresponding to variations in the rER and zymogen granules.     

Cell proliferation kinetics of epidermis and sebaceous glands in relation to chalone action.

Median S-phase lengths of pinna epidermis and sebaceous glands, and of epithelia from the oesophagus and under surface of the tongue of Albino Swiss S mice were estimated by the percentage labelled mitoses method (PLM). The 18.4 and 18,8 hr for the median length of S-phase for pinna epidermis and sebaceous glands respectively made it possible for these two tissues to be used experimentally for testing tissue specificity in chalone assay experiments. The 10.0 and 11.5 hr for oesophagus ang tongue epithelium respectively made experimental design for chalone assay difficult when pinna epidermis was the target tissue. The results of the Labelling Index measured each hour throughout a 24-hr period showed no distinct single peaked diurnal rhythm for pinna epidermis and sebaceous glands. Instead a circadian rhythm with several small peaks occurred which would be expected if an S-phase of approximately 18 hr was imposed on the diurnal rhythm. This indicates that there may be very little change in the rate of DNA synthesis. The results are given for the assay in vivo of purified epidermal G1 and G2 chalones, and the 72--81% ethanol precipitate of pig skin from which they could be isolated. These experiments were performed over a time period which took into account the diurnal rhythm of activity of the mice as well as the S-phase lengths. Extrapolating the results with time of action of the chalone shows that the G1 chalone acts at the point of entry into DNA synthesis and that the S-phase length was approximately 17 hr for both the pinna epidermis and sebaceous glands. This may be a more correct value since the PLM method overestimates the median S-phase length as it is known that in pinna skin the [3H]TdR is available to the tissues for 2 hr and true flash labelling does not take place. The previous reports that epidermal G1 chalone acts some hours prior to entry into S-phase resulted from experiments on back skin where the S-phase is shorter and there is a pronounced diurnal rhythm which could mask the chalone effect. The epidermal G2 chalone had no effect on DNA synthesis even at different times in the circadian rhythm. Thus the circadian rhythms and S-phase lengths of the test tissues need to be considered when experiments are performed with chalones. Ideally, the target tissues selected for cell line specificity tests should have the same cell kinetics for the easier and more accurate assessment and interpretation of results. When the tissues have markedly different cell kinetics, experimental procedures and results need to be evaluated accordingly. The point of action of G1 chalone can only be assessed if the effect is measured over the peak of incorporation of [3H]TdR into DNA. The results of the effects of skin extracts are analysed in relation to changes in the availability of [3H]TdR for the incorporation into DNA and to the possibility of there being two distinct populations of proliferating cells.     

Circadian rhythm of DNA synthesis and mitotic activity in tongue keratinocytes

Tongue keratinocytes have a high mitotic index (MI) with an evident circadian variation. Our study set out to compare and contrast two phases of the cell cycle: DNA synthesis (S-phase), with inmunocytochemical detection by bromodeoxyuridine (BrdU), and mitosis (M-phase), by the colchicine-arrest of metaphase method, exploring both the dorsal and ventral surfaces of the mouse tongue throughout a circadian period. Adult male mice standardized for light periodicity used for MI experiment were injected intraperitoneally with colchicine. Other animals were injected intraperitoneally with 5-BrdU for S-phase determination. Animals given both treatments were divided into six groups and killed at 4 h intervals until 20:00 h. Tongue samples were processed for histology and immuno-histochemistry. S and M indices were expressed as labelled nuclei or colchicine metaphases, respectively, per 1000 nuclei. Peak MI occurred at 12:00, with the minimum value at 20:00 on dorsal and ventral tongue surfaces. Peak S-phase was at 04:00, whereas the minimum value was at 16:00 for both surfaces. These results show that the proliferative activity of the tongue epithelium is of similar intensity and temporal distribution on both surfaces.     

CIRCADIAN INFLUENCE ON THE WAVE FORM OF THE FREQUENCY OF LABELED MITOSES IN MOUSE CORNEAL EPITHELIUM

The FLM was studied in mouse corneal epithelium at two different phases of the mouse circadian system. The following results were obtained: (1) a circadian rhythm was found in the corneal mitotic index of both experimental groups, with the peak near 09.00 hours, the trough near 21.00 hours and a 10-fold difference in values between the peak and the trough; (2) when ³H-TdR was injected at 09.00 hours, the first wave of labelled mitoses rose earlier, attained a broader peak and fell later when compared to the first wave of labeled mitoses obtained when ³H-TdR was injected at 21.00 hours; and (3) in both experimental groups there was a suggestion of a second wave of labeled mitoses, especially in the group injected with ³H-TdR at 21.00 hours, where a flat second wave appeared in the region of 60–90 hr. Transit times derived from FLM curves obtained from such synchronous populations of cells are of questionable validity. Circadian rhythmicity and other causes of synchronization must be ruled out before accepting a single FLM curve as a valid indication of transit times.     

Diurnal Profiles of Blood Glucose in Relation to Time of Administration of Melatonin in Male Spotted Munia (Lonchura punctulata)

The study was aimed at demonstration of the effect of a single acute dose of melatonin (0.5 mg/ 100 g body wt.) on the diurnal profile of blood glucose in male spotted munia in relation to the administration of hormone at the onset of light (i.e., at 06.00 h) or at the onset of darkness (i.e., at 18.00 h) under natural photoperiodic (~12L : 12D) conditions. Blood samples from all birds belonging to the control, sham-control (administered only with the vehicle of hormone, i.e., ethanol-saline 1:9 v/v), and melatonin treated groups were collected at four time points, i.e. 06.00 h, 12.00 h, 18.00 h, and 24.00 h, in a 24 hour cycle. The blood glucose levels in control and sham-control birds showed marked variation with regard to the time of sampling, with a mid-day peak and morning nadir. Exogenous melatonin induced a significant alteration in this diurnal pattern of blood glucose with a marked variation in relation to the time of administration of melatonin. While morning administration of melatonin resulted in hypoglycemia at 12.00 h and 24.00 h and hyperglycemia at 18.00 h, the response to evening injection of melatonin was only hypoglycemic at 24.00 h leaving the glycemic values at other time-points almost unaltered compared to the blood glucose levels in control and sham-control munias. The results of this investigation demonstrate for the first time that a schedule of morning administration of melatonin induces a more broad range of variations in the blood glucose levels than a schedule of evening administration does.     

CELL PROLIFERATION KINETICS OF EPIDERMIS AND SEBACEOUS GLANDS IN RELATION TO CHALONE ACTION

Median S-phase lengths of pinna epidermis and sebaceous glands, and of epithelia from the oesophagus and under surface of the tongue of Albino Swiss S mice were estimated by the percentage labelled mitoses method (PLM). The 18.4 and 18.8 hr for the median length of S-phase for pinna epidermis and sebaceous glands respectively made it possible for these two tissues to be used experimentally for testing tissue specificity in chalone assay experiments. The 10.0 and 11.5 hr for oesophagus and tongue epithelium respectively made experimental design for chalone assay difficult when pinna epidermis was the target tissue. The results of the Labelling Index measured each hour throughout a 24-hr period showed no distinct single peaked diurnal rhythm for pinna epidermis and sebaceous glands. Instead a circadian rhythm with several small peaks occurred which would be expected if an S-phase of approximately 18 hr was imposed on the diurnal rhythm. This indicates that there may be very little change in the rate of DNA synthesis. The results are given for the assay in vivo of purified epidermal G₁ and G₂ chalones, and the 72–81% ethanol precipitate of pig skin from which they could be isolated. These experiments were performed over a time period which took into account the diurnal rhythm of activity of the mice as well as the S-phase lengths. Extrapolating the results with time of action of the chalone shows that the G₁ chalone acts at the point of entry into DNA synthesis and that the S-phase length was approximately 17 hr for both the pinna epidermis and sebaceous glands. This may be a more correct value since the PLM method overestimates the median S-phase length as it is known that in pinna skin the [³H]TdR is available to the tissues for 2 hr and true flash labelling does not take place. The previous reports that epidermal G₁ chalone acts some hours prior to entry into S-phase resulted from experiments on back skin where the S-phase is shorter and there is a pronounceddiurnal rhythm which could mask the chalone effect. The epidermal G, chalone had no effect on DNA synthesis even at different times in the circadian rhythm. Thus the circadian rhythms and S-phase lengths of the test tissues need to be considered when experiments are performed with chalones. Ideally, the target tissues selected for cell line specificity tests should have the same cell kinetics for the easier and more accurate assessment and interpretation of results. When the tissues have markedly different cell kinetics, experimental procedures and results need to be evaluated accordingly. The point of action of G, chalone can only be assessed if the effect is measured over the peak of incorporation of 13H]TdR into DNA. The results of the effects of skin extracts are analysed in relation to changes in the availability of i3H]TdR for the incorporation into DNA and to the possibility of there being two distinct populations of proliferating cells.     

Diurnal Profiles of Blood Glucose in Relation to Time of Administration of Melatonin in Male Spotted Munia (Lonchura punctulata)

The study was aimed at demonstration of the effect of a single acute dose of melatonin (0.5 mg/ 100 g body wt.) on the diurnal profile of blood glucose in male spotted munia in relation to the administration of hormone at the onset of light (i.e., at 06.00 h) or at the onset of darkness (i.e., at 18.00 h) under natural photoperiodic (~12L : 12D) conditions. Blood samples from all birds belonging to the control, sham-control (administered only with the vehicle of hormone, i.e., ethanol-saline 1:9 v/v), and melatonin treated groups were collected at four time points, i.e. 06.00 h, 12.00 h, 18.00 h, and 24.00 h, in a 24 hour cycle. The blood glucose levels in control and sham-control birds showed marked variation with regard to the time of sampling, with a mid-day peak and morning nadir. Exogenous melatonin induced a significant alteration in this diurnal pattern of blood glucose with a marked variation in relation to the time of administration of melatonin. While morning administration of melatonin resulted in hypoglycemia at 12.00 h and 24.00 h and hyperglycemia at 18.00 h, the response to evening injection of melatonin was only hypoglycemic at 24.00 h leaving the glycemic values at other time-points almost unaltered compared to the blood glucose levels in control and sham-control munias. The results of this investigation demonstrate for the first time that a schedule of morning administration of melatonin induces a more broad range of variations in the blood glucose levels than a schedule of evening administration does.     

Circadian oscillatory patterns of oxygen uptake in individual workers of the ant Camponotus rufipes

Abstract. Respiratory rates of individual workers of Camponotus rufipes Fabricius (Hymenoptera: Formicidae) were measured at 25C and LD 12:12 h (lights on 06.00 hours), DL 12:12h (lights on 18.00 hours), LL (850 lux) and DD (red light, 20–30 lux), using the micro-Warburg technique. Worker ants were collected from natural nest during the winter of 1987 in a woodland park in the region of Rio Claro, Sāo Paulo, Brazil. The respiration of ants showed a circadian rhythm with acrophase ranging from 20 h 41 min to O1 h 18 min and from 10 h 32 min to 12 h 22 min at LD and DD, respectively. In constant darkness the rhythmometric variables were similar to those presented by ants kept at LD 12:12 h. Under constant light no circadian rhythm in the respiration rates was found. A reduction in the amplitude was observed, indicating an inhibitory effect of this light regime on the respiration process.     

IgE-mediated mast cell activation induces Langerhans cell migration in vivo

Langerhans cells and mast cells are both resident in large numbers in the skin and act as sentinel cells in host defense. The ability of mast cells to induce Langerhans cell migration from the skin to the draining lymph node in vivo was examined. Genetically mast cell-deficient (W/Wv) mice and control mice were sensitized with IgE Ab in the ear pinna. Seven to 14 days later, mice were challenged with Ag i.v. After a further 18-24 h, epidermal sheets and draining auricular lymph nodes were examined using Langerin/CD207 immunostaining. In mast cell-containing mice, a significant decrease in the number of Langerhans cells was observed at epidermal sites of mast cell activation. A significant increase in total cellularity and accumulation of Langerin-positive dendritic cells was observed in the auricular lymph nodes, draining the sites of IgE-mediated mast cell activation. These changes were not observed in W/Wv mice, but were restored by local mast cell reconstitution. Treatment of mast cell-containing mice with the H2 receptor antagonist cimetidine significantly inhibited the observed IgE/Ag-induced changes in Langerhans cell location. In contrast, Langerhans cell migration in response to LPS challenge was not mast cell dependent. These data directly demonstrate the ability of mast cells to induce dendritic cell migration to lymph nodes following IgE-mediated activation in vivo by a histamine-dependent mechanism.     

Acinar cell proliferation in the parotid and submandibular salivary glands of the neonatal rat

Abstract. Although the rat salivary glands are deficient in acini at birth, acinar cells proliferate rapidly during the early post-natal period. The pattern of acinar cell proliferation was analysed in the parotid and submandibular glands of neonatal rats from day of birth until day 34. Mitotic and [³H]thymidine ([³H]TdR) labelling indices of the two glands show distinctly different patterns. Analysis of cell division in the rat parotid gland demonstrated a peak of mitotic index at 14 days (2.9 ± 0.4%) and labelling index at 16 days (25.2 ± 2.1%). Submandibular gland acinar cell proliferation reaches a zenith between 7–8 days; labelling index (14.2 ± 1.1%) and mitotic index (2.3 ± 0.3%). Cell proliferation decreases rapidly in both glands after reaching a peak in activity. Gland size increases more rapidly in the submandibular gland which correlates with the earlier shift from cell proliferation to differentiation which occurs in this organ. Circadian rhythms of [³H]TdR incorporation were also investigated in this study. A circadian rhythm of [³H]TdR incorporation into DNA occurs at 15 days after birth with a peak at 06.00 hours in both glands and a trough occurring at 15.00 hours in parotid gland and 18.00 hours in the submandibular gland. Determination of specific activity of DNA (ct/min per μg DNA) on days 8, 10, 12, 13, 14, 15, and 16 after birth at 06.00 and 15.00 hours indicated that a circadian rhythm in [³H]TdR incorporation into DNA began on day 14. The developmental switch from suckling to solid food may be an initiating factor in the sychronization of the circadian rhythm in cell proliferation.     

Chronopharmacology of pancuronium in rats anesthetized by CT 1341 (Alfatésine)]

A circadian variation in the curarizing ability of pancuronium bromide (Pavulon) has been documented in an homogenous group of 100 Wistar AF-SPF adult male rats anaesthetized by the steroid anaesthetic althesin (Alfatésine, CT 1341). Animals were maintained at 24 +/- 2 degrees C and synchronized with natural light 06.00 to 18.00 and darkness (october 1977). We observed significant circadian rhythms for both of these agents: first the induction of anaesthesia by althesin was markedly pronounced at 15.30 and varied with season. Secondly the maximum effect of pancuronium was recorded at 08.00. When rats were anaesthetized by sodium pentobarbital under similar experimental conditions but during a different season (january to may) we observed a similar circadian rhythm for pancuronium. These data indicate that: a) the type of anaesthesia used in the protocol may not be of importance in demonstration of a curarizing rhythm in the rat but, b) the possibility of a seasonal component being present and effecting this rhythm needs to be investigated.     

Night-time behavior of stabled and pastured peri-parturient ponies

Pregnant and lactating pony mares were observed in two environments, stable and pasture. Twenty-six pony mares were observed on pasture for 2 weeks before and after parturition. The behavior of each mare was recorded every 30 min from 18.00 to 06.00 h. The mutually exclusive behaviors were standing (either standing alert or standing at rest with a hindlimb flexed), grazing (prehending or masticating grass), walking, lying in sternal recumbency and lying in lateral recumbency. The total time-budget for prepartum mares on pasture was 55.3 ± 4.1% grazing, 32.9 ± 3.3% standing, 6.0 ± 1.5% lying in sternal recumbency, 2.7 ± 0.7% walking and 1.4 ± 0.6% lying in lateral recumbency. Grazing and standing occurred at all times, but grazing was most common from 18.00 to 21.00 h and after 05.00 h. Lying was most common between 01.00 and 04.00. Lying in lateral recumbency occurred only after dark, in pre-partum mares. The total time-budget for post-partum mares on pasture was 68.6 ± 4.0% grazing, 22.5 ± 3.0% standing, 4.7 ± 1.0% walking, 4.2 ± 1.2% lying in sternal recumbency and 0.2 ± 0.2% lying in lateral recumbency. Lying in lateral recumbency was seen only at 18.00 h. Lying in sternal recumbency occurred between 21.00 and 04.30 h. More time was spent grazing by the post-partum mares than by the pre-partum mares.The same behaviors were recorded for stabled pony mares except that eating hay rather than grazing constituted the ingestive behavior quantified. The total time-budget for pregnant stabled ponies was 71 ± 3% standing, 15 ± 3% eating, 0.5 ± 0% lying in lateral recumbency, 0.5 ± 0.2% walking and 12.1 ± 2.3% lying in sternal recumbency. Eating decreased and standing increased during the night. Most lying was seen between 01.30 and 05.00 h. Lying in lateral recumbency occurred between 19.30 and 03.30 h. The total nocturnal time-budget of post-partum stalled ponies was 67 ± 3% eating, 19 ± 3% standing and 13.0 ± 2.3% lying in sternal recumbency. Post-partum mares were not observed to walk or to lie in lateral recumbency.The change in behavior after parturition may reflect: (1) nutritional demands of lactation; (2) maternal protective behavior; (3) response to seasonal changes in the environment.