Seasonal variation of body weight loss after bariatric surgery

Seasonal variations have been described in humans in several variables such as sleep, mood, appetite, food preferences, or body weight. We hypothesized that these variations could also influence the decrease in body weight rate in patients submitted to body weight loss interventions. Thus, here we tested the variations of weight loss according to the time of the year the surgery took place in a group patients (n = 1322) submitted to bariatric surgery in the Hospital Universitari de la Vall d’Hebron in Barcelona (geographical coordinates: 41°25?41″N 2°8?32″E). For the analysis, the percentage of total body weight loss (%TWL), excess body weight loss (%EWL) and percentage of body mass index loss (%BMIL) were calculated at 3 (n = 1255), 6 (n = 1172), 9 (n = 1002), and 12 months (n = 1076) after surgery. For %EWL and %BMIL a statistically significant seasonal variation was detected when the variables were calculated at 3 months, but not at the other times, with more weight loss in summer-fall. However, seasonal variations were not detected for %TWL (p = 0.09). The mean amplitude of the seasonal rhythm for %EWL was of 1.8%, while for the rhythm of %BMIL was 0.7%. Moreover, a second peak was detected in January–February modulating the seasonal rhythm of the two variables. Results confirm seasonal variations in humans and indicate that short term responses to weight loss can be modulated by the time of year.     

Circadian and circannual study of the hypophyseal-gonadal axis in healthy young males

Circadian and circannual variations of Testosterone, FSH and LH secretions, other than Oral Body Temperature (OBT) have been studied in four healthy males. OBT showed a constant circadian rhythm with an acrophase located in the afternoon. Plasma Testosterone exhibited both a circadian (acrophase = hr 09,28) and a circannual rhythm (acrophase = 22 february); plasma FSH also showed a circannual rhythm (acrophase = 13 february). By mean chronogram +/- SEM we documented the highest LH levels in December and the lowest in February. These observations would suggest the hypothesis that the winter could be the period in which the hypophysis-gonadal axis in young males exhibits its maximal activity as previously documented for other hormones.     

Seasonal variation in sleeping metabolic rate,thyroid activity,and leptin

We investigated seasonal variation in sleeping metabolic rate (SMR) and the possible relation to body composition, thyroid activity, and leptin. Twenty-five healthy volunteers were examined four times during the year: in spring (April, May), summer (July, August), autumn (October, November), and winter (January, February). Body composition was determined using a three-compartment model based on underwater weighing and the deuterium dilution method. SMR was measured during an overnight stay in a respiration chamber. A blood sample was taken for the analysis of free and total thyroxine, TSH, and leptin. SMR showed a significant seasonal variation (P < 0.01) with a maximum in winter (4.54 kJ/min) and a minimum in summer (4.34 kJ/min). The amplitude was 0.10 +/- 0.02 kJ/min, and the phase was November 5th. Season explained 17% of the intraindividual variation in SMR. The circannual rhythm in SMR could not be explained by changes in body composition, thyroid activity, or leptin. Interindividual variation in SMR was explained by fat-free mass (P < 0.001) and leptin (P < 0.001).     

Circannual variation of efficacy outcomes in patients with newly diagnosed metastatic colorectal cancer and treated with first-line chemotherapy

Seasonal variation of baseline diagnosis (or clinical suspect) of stage I–III colorectal cancer patients has been repeatedly reported as an independent variable influencing overall survival. However, data are conflicting and no information is available about such a rhythm in advanced stage patients. To test whether a circannual rhythm of efficacy outcomes can be detected in this setting, we collected data about response rate (RR), progression-free survival (PFS), and overall survival (OS) to first-line chemotherapy of 1610 newly diagnosed metastatic patients treated at four independent centers. Responses to first-line chemotherapy were available for 1495 patients. A strong circannual rhythm in RR was evident, with the higher proportion of responding patients in the subgroup diagnosed in January (acrophase). At the time of data cutoff, 1322 patients progressed and 986 died, with median PFS and OS of 11 and 25.6 months, respectively. A circannual rhythmicity of the proportion of patients progressing at 6 months and surviving at 1 year was demonstrated, with acrophases located both in winter (February and January, respectively), similar to what reported for RR. Several interpretations about the genesis of this cyclic variation could be claimed: the rhythm in sunlight exposure and, as a consequence, of vitamin D serum levels and folate degradation, the variability in toxic effect intensity of chemotherapy, and the rhythm in the biological behavior of tumor cells. This observation is worth of further investigation both in preclinical and in clinical settings in order to better elucidate the underlying mechanisms.     

Influence of pregnancy on diurnal and seasonal changes in cortisol, T3 and T4 levels in the mare blood serum.

1. The diurnal changes in the level of total protein, cortisol, T3 and T4 were studied in four barren and four pregnant standard-bred mares, kept and examined under the same conditions. 2. Blood samples were taken every 4 hr. for one day each month, throughout one year. 3. In barren mares, a diurnal rhythm in cortisol level (acrophase at 0530 hr in summer and at 0830 hr in winter) and in T3 level (acrophase at 1330 hr in summer and at 1800 hr in winter) was found. 4. In pregnant mares, a diurnal rhythm in cortisol level only till 5th month of pregnancy was observed. 5. A diurnal rhythm in T3 level was found throughout the pregnancy, with acrophase always at 1400 hr. 6. No diurnal rhythm in the total protein content and in the T4 level was observed. 7. In both groups of mares the seasonal cyclicity in T3 and T4 levels were found. A seasonal cyclicity in cortisol level was found only in pregnant mares. 8. Pregnancy abolished seasonal cyclicity in total protein and showed it in cortisol level. 9. Pregnancy in mares modifies diurnal rhythms as well as seasonal cycles in secretion and metabolism of the hormones studied.     

Importance of photoperiodic signal quality to entrainment of the circannual reproductive rhythm of the ewe

An endogenous circannual rhythm drives the seasonal reproductive cycle of a broad spectrum of species. This rhythm is synchronized to the seasons (i.e., entrained) by photoperiod, which acts by regulating the circadian pattern of melatonin secretion from the pineal gland. Prior work has revealed that melatonin patterns secreted in spring/summer entrain the circannual rhythm of reproductive neuroendocrine activity in sheep, whereas secretions in winter do not. The goal of this study was to determine if inability of the winter-melatonin pattern to entrain the rhythm is due to the specific melatonin pattern secreted in winter or to the stage of the circannual rhythm at that time of year. Either a summer- or a winter-melatonin pattern was infused for 70 days into pinealectomized ewes, centered around the summer solstice, when an effective stimulus readily entrains the rhythm. The ewes were ovariectomized and treated with constant-release estradiol implants, and circannual cycles of reproductive neuroendocrine activity were monitored by serum LH concentrations. Only the summer-melatonin pattern entrained the circannual reproductive rhythm. The inability of the winter pattern to do so indicates that the mere presence of a circadian melatonin pattern, in itself, is insufficient for entrainment. Rather, the characteristics of the melatonin pattern, in particular a pattern that mimics the photoperiodic signals of summer, determines entrainment of the circannual rhythm of reproductive neuroendocrine activity in the ewe.     

Circannual rhythm of plasma thyrotropin in middle-aged and old euthyroid subjects

The circannual rhythm of plasma thyrotropin (TSH) was evaluated in 8,310 euthyroid, serially independent, young, middle-aged and old men and women. A statistically significant circannual rhythm of plasma TSH was validated, by the mean group-cosinor method, in the middle-aged and old men and women (p less than 0.05), with acrophase in December, whereas the young subjects did not show any rhythm. No significant correlation was found between TSH plasma levels and free thyroxine (fT4) or ambient temperature in any group. Moreover, plasma fT4 did not show seasonal variations.     

Circannual Incidence of Giardia Lamblia

A circannual rhythm of Giardia lamblia positive stools was found by examination of records from three clinical laboratories in central Arkansas for the period 1980-1986. Cosinor analysis of monthly Giardia incidence based on stool specimen records from approximately 12,000 patients over the 7-year period revealed a circannual rhythm (P< 0.001) on the basis of percent positive patients/month, with a computive acrophase occurring in late summer and minimum values in the winter. Patients involved in the study were primarily from the central Arkansas metropolitan areas, southern delta regions and northern mountainous regions of the state. Analysis of the data on the basis of total positive Giardia patients/month also revealed a circannual rhythm with the acrophase again occurring in late summer. The overall mean for percent positive stool specimens for the 7-year period was 5.3%, compared with the national average of 3.8% for G. lamblia positive stools. The data indicate that there may be a “Giardia season” in Arkansas since they could not be explained on the basis of day-care age distribution, or geographic origin. Awareness by epidemiologists, public health officials and other health care professionals of this circannual incidence of giardiasis is important for the prevention, diagnosis and treatment of this infectious disorder.     

Circannual incidence of Giardia lamblia

A circannual rhythm of Giardia lamblia positive stools was found by examination of records from three clinical laboratories in central Arkansas for the period 1980-1986. Cosinor analysis of monthly Giardia incidence based on stool specimen records from approximately 12,000 patients over the 7-year period revealed a circannual rhythm (P less than 0.001) on the basis of percent positive patients/month, with a computive acrophase occurring in late summer and minimum values in the winter. Patients involved in the study were primarily from the central Arkansas metropolitan areas, southern delta regions and northern mountainous regions of the state. Analysis of the data on the basis of total positive Giardia patients/month also revealed a circannual rhythm with the acrophase again occurring in late summer. The overall mean for percent positive stool specimens for the 7-year period was 5.3%, compared with the national average of 3.8% for G. lamblia positive stools. The data indicate that there may be a "Giardia season" in Arkansas since they could not be explained on the basis of day-care age distribution, or geographic origin. Awareness by epidemiologists, public health officials and other health care professionals of this circannual incidence of giardiasis is important for the prevention, diagnosis and treatment of this infectious disorder.     

Different behavior of body temperature and total locomotor activity daily rhythms during light/dark cycle in stabled Oryctolagus cuniculus

In order to investigate the potential causal link between the rhythm of activity and body temperature, we simultaneously recorded rectal temperature and total locomotor activity in five clinically healthy female rabbits (blue Vienna breed), 12 week old and mean body weight 2.7 ± 0.3. Animals were housed in individual cages (90?×?50?×?35 cm) under natural 12/12 light/dark cycle. Total locomotor activity was monitored for 15 days by an activity data logger. On day 1, 5, 10, and 15 rectal temperature was recorded every 2 h for a 24-h period. Application of single cosinor method showed a nocturnal daily rhythm of rectal temperature with a range of oscillation of about 1 °C, acrophase after dusk and low robustness value. The daily rhythm of locomotor activity showed its acrophase in the middle of the scotophase and a high robustness value. This information improves the knowledge available on the circadian biology of rabbits useful in the evaluation of physiology of this species.     

Rates of insufficiency and deficiency of vitamin D levels in elite professional male and female skiers: A chronobiologic approach

Vitamin D is essential for the maintenance and promotion of musculoskeletal health, for the functioning of the immune, cardiovascular and reproductive systems, and its main action is to keep calcium and phosphate plasmatic physiological concentrations at intestinal, renal and bony level. Vitamin D affects several parameters related to physical performance too and a particularly high percentage of vitamin D insufficiency and deficiency in professional athletes has been observed. Several variables are able to impair the synthesis of 25(OH)D in athletes, specifically both genetic and environmental factors, but the most probable explanation for the deficient/insufficient vitamin D levels is the insufficient ultraviolet B light (UVB) exposure during winter. To confirm this, the existence of a circannual rhythm of vitamin D in professional soccer players, highlighting a peak in summer and lowest values in winter regardless the period of the season, has been documented. Nonetheless, from what we are aware of, no other study adopted a chronobiologic approach to better understand and describe the circannual variations of serum 25(OH)D in other sport disciplines. Therefore, we studied serum vitamin D in a cohort of top-level professional skiers, during a period of three consecutive competitive seasons (2015, 2016 and 2017), in order to evaluate, with a rhythmometric approach, the vitamin D behavior along the year. The study population was composed by 152 professional Italian alpine skiers of FISI (Winter Sport Italian Federation), 63 females and 89 males (mean age: 24.1 ± 3.2 years) and a total of 298 blood drawings were carried out to determine plasma 25(OH)D. Vitamin D data were compared between genders and then processed with the population mean cosinor tests to evaluate the presence of a circannual rhythm, both for female and male athletes. In total, 77 skiers (50.7%) showed, at least once during the three competitive seasons, an insufficient level of 25(OH)D and other 45 subjects (29.6%) showed a deficient status; no differences were observed between genders (mean for females: 26.9 ± 8.1 ng/mL; mean for males: 27.4 ± 7.6 ng/mL). In addition, the rhythmometric analysis highlighted the existence of a significant circannual rhythm for both female and male professional skiers; the acrophases (Φ) occurred in July and both MESOR (M) and amplitude (A) were comparable between the two groups. Our data indicate that, despite the physical effort spent, vitamin D follows a classical season-associated rhythm with a peak in summer and a nadir in winter. Moreover, the percentage of insufficiency and deficiency is in line with that of the general population. In conclusion, our findings reinforce the hypothesis that there is no direct effect of physical activity on vitamin D metabolism and that the factors involved in the determination of vitamin D levels in the general population are valid also for athletes.     

Rhythms in human bone marrow and blood cells

In 24h studies of bone marrow (BM), circadian stage-dependent variations were demonstrated in the proliferative activity of BM cells from subsets of 35 healthy diurnally active men. On an average, the percentage of total BM cells in deoxyribonucleic acid (DNA) synthesis phase was 188% greater at midday than at midnight (circadian rhythm: p = 0.018; acrophase or peak time of 13: 16h). Patients with malignant disease (n = 15) and a normal cortisol circadian rhythm showed higher fractions of BM cells in S-phase at midday. Colony-forming units--granulocyte/macrophage (CFU-GM), an indicator of myeloid progenitor cells, showed the same circadian variation as DNA S-phase (average range of change or ROC = 136%; circadian rhythm: p < 0.001; acrophase of 12:09h). Deoxyribonucleic acid S-phase and CFU-GM in BM both showed a circannual rhythm (p = 0.015 and 0.008) with an identical acrophase of August 12. The daily peak in BM glutathione content, a tripeptide involved in cellular defense against cytotoxic damage, preceded BM proliferative peaks by 4-5 h (ROC = 31-90%; circadian rhythm: p = 0.05; acrophase of 08:30h). Myeloid (ROC = 57%; circadian rhythm: p = 0.056; acrophase at 08:40h) and erythroid (ROC = 26%; circadian rhythm: p = 0.01; acrophase of 13:01h) precursor cells were positively correlated (r = 0.41; p < 0.001), indicating a circadian temporal relationship and equal influence on S-phase of total BM cells. Yield of positive selected CD34+ progenitor stem cells also showed significant circadian variation (ROC = 595%; circadian rhythm: p = 0.02; acrophase of 12:40h). Thus, the temporal synchrony in cell cycling renders BM cells more sensitive at specific times to hematopoietic growth factors and cell cycle-specific cytotoxic drugs. Moreover, proper timing of BM harvesting may improve progenitor cell yield. When using marker rhythms in the blood to allow for individualized timing of BM procedures, the times of low values in white blood corpuscles, neutrophils, and lymphocytes and high values in cortisol were predictive of the times of highest BM erythroid, myeloid, and total S-phase numbers occurring in the following 12 h.     

Circannual versus seasonal variations of longitudinally sampled 25-hydroxycholecalciferol serum levels

Seasonal variations in human serum levels of 25-hydroxycholecalciferol (25OHD) have been largely documented in transverse studies of population. But seasonality is not per se a demonstration that 25OHD serum levels fluctuate along the course of year according to a waveform profile with a periodic rhythm. Because of this, we attempted to investigate the possible occurrence of a circannual rhythm for 25OHD serum levels in a longitudinal design, by fitting a 365.25-day cosine curve to temporal biodata recorded in 10 clinically healthy subjects, monthly sampled for RIA determinations of 25OHD. Cosinor procedure statistically validated the occurrence of a circannual rhythm for 25OHD serum concentrations at a highly significant level of probability (P = 0.0015) for null hypothesis amplitude = 0. With 95% of probability, amplitude ranges from 5.0 to 16.5 ng/ml (mean value of oscillation = 10 ng/ml), while acrophase is temporally located from September 14 to December 3 (mean timing = October 21). Yearly, mean values for 25OHD serum concentrations is of 40.3 +/- 5.4 ng/ml as quantified by the line which transversely divides the cosine curve interpolating original biodata. By calculating the band of a complete 12 months variability which includes 90% of the distribution with 90% confidence limits, the circannual chronodesm of 25OHD serum levels has been obtained. Such a chronodesmic sinusoid has been compared to the circannual chronogram. By this comparison, a dissociation between the crest (October) and the peak (August) has been detected. The finding suggests that seasonal variations are superimposed to the circannual rhythm. Seasonal but also circannual changes, thus, characterize the yearly variability of 25OHD serum levels in man.     

Circannual rhythm of plasmatic vitamin D levels and the association with markers of psychophysical stress in a cohort of Italian professional soccer players

Adequate plasmatic Vitamin D levels are crucial to maintain calcium homeostasis and bone metabolism both in the general population and in athletes. Correct dietary supply and a regular sun exposure are fundamental for allowing the desired and effective fitness level. Past studies highlighted a scenario of Vitamin D insufficiency among professional soccer players in several countries, especially in North Europe, whilst a real deficiency in athletes is rare. The typical seasonal fluctuations of Vitamin D are wrongly described transversally in athletes belonging to teams that play at different latitudes and a chronobiologic approach studying the Vitamin D circannual rhythm in soccer players has not been described yet. Therefore, we studied plasma vitamin D, cortisol, testosterone, and creatin kinase (CK) concentrations in three different Italian professional teams training at the same latitude during a period of two consecutive competitive seasons (2013 and 2014). In this retrospective observational study, 167 professional soccer players were recruited (mean age at sampling 25.1 ± 4.7 years) and a total of 667 blood drawings were carried out to determine plasma 25(OH)D, serum cortisol, serum testosterone and CK levels. Testosterone to cortisol ratio (TC) was calculated based as a surrogate marker of overtraining and psychophysical stress and each athlete was drawn until a maximum of 5 times per season. Data extracted by a subgroup of players that underwent at least 4 sample drawings along a year (N = 45) were processed with the single and population mean cosinor tests to evaluate the presence of circannual rhythms: the amplitude (A), acrophase (Φ) and the MESOR (M) are described. In total, 55 players (32.9%) had an insufficient level of 25(OH)D during the seasons and other 15 athletes (9.0%) showed, at least once, a deficiency status of Vitamin D. The rhythmometric analyses applied to the data of Vitamin D revealed the presence of a significant circannual rhythm (p < 0.001) with the acrophase that occurred in August; the rhythms of Vitamin D levels were not different neither among the three soccer teams nor between competitive seasons. Cortisol, testosterone and TC showed significant circannual rhythms (p < 0.001): cortisol registered an acrophase during winter (February) while testosterone and TC registered their peaks in the summer months (July). On the contrary, CK did not display any seasonal fluctuations. In addition, we observed weak but significant correlations between 25(OH)D versus testosterone (r = 0.29 and p < 0.001), cortisol (r = ?0.27 and p < 0.001) and TC (r = 0.37 and p < 0.001). No correlation was detected between Vitamin D and CK. In conclusion, the correct chronobiologic approach in the study of annual variations of Vitamin D, cortisol and testosterone could be decisive in the development of more specific supplementation and injury prevention strategies by athletic trainers and physicians.     

Circannual variation of intestinal cell proliferation in BDF1 male mice on three lighting regimens

BDF, male mice were studied over a 24-hr span in winter, spring, summer and fall. For three weeks prior to study, one-third of the animals were kept under a lighting regimen of 8 hr light alternating with 16 hr of darkness (LD 8:16), one-third on a lighting regimen of LD 12:12 and a remainder on a lighting regimen of LD 16:8. During each study, subgroups of animals on all three lighting regimens were killed at 4-hr intervals over a 24-hr span. Twenty minutes prior to being killed, the animals received 5yCi of [³Hthymidine/0.2 ml/20 gm of body weight intraperitoneally. The thymidine uptake in the DNA of the colon and of the small intestine were studied as an index of cell proliferation. A circadian rhythm in [³H]-thymiduie uptake in the colon was found and validated by cosinor analysis. This rhythm was similar in acrophase and amplitude in the animals kept on LD 8:16 and LD 12:12. Also in the mice on LD 16:8, there was a statistically significant circadian rhythm of ('HJ-thymidtne uptake in the DNA of the colon during all four seasons. The acrophases of this rhythm, however, varied widely suggesting free running. A circadian rhythm of pHJ-thymidine uptake in small intestine was less consistent. In animals on all three lighting regimens, however, a circannual variation of f'HJ-thymidine uptake in DNA in colon and small intestine was found with the highest uptake during summer. This study indicates that a lighting regimen of LD 16:8 does not reliably synchronize the circadian rhythm of [³H]-thymidine uptake in the colon. It further shows a circannual rhythm of this function in the colon and in the small intestine which persists under three lighting regimens (LD 8:16, 12:12 and 16:8) maintained for three to four weeks prior to being killed.     

Circannual Variations in Blood Cholesterol Levels

The seasonality of blood cholesterol is still not well established. Some have described a seasonal pattern with highest levels during autumn and lowest in summer, whereas others have reported no change. A number of studies showed circannual variations with maximum levels in winter and minimum in summer. The aim of this study was to examine the circannual variation of cholesterol in a large cohort in Israel. In the Israeli CORDIS study, employees of 21 factories were screened for cardiovascular disease risk factors during 1985-7. As part of the information gathered, serum cholesterol and blood counts were available for 3,726 men and 1,514 women. Serum cholesterol levels fit a circannual rhythm assessed by the cosinor analysis. Highest levels of serum cholesterol were found in spring and lowest in summer. We conclude that there is a circannual change of serum cholesterol, which could be partially associated with changes in environmental temperature. The circannual variation in serum cholesterol was considerable and should be taken into account when carrying out clinical evaluation of patients.     

Circannual Variation of Intestinal Cell Proliferation in Bdf1 Male Mice on Three Lighting Regimens

BDF, male mice were studied over a 24-hr span in winter, spring, summer and fall. For three weeks prior to study, one-third of the animals were kept under a lighting regimen of 8 hr light alternating with 16 hr of darkness (LD 8:16), one-third on a lighting regimen of LD 12:12 and a remainder on a lighting regimen of LD 16:8. During each study, subgroups of animals on all three lighting regimens were killed at 4-hr intervals over a 24-hr span. Twenty minutes prior to being killed, the animals received 5yCi of [³Hthymidine/0.2 ml/20 gm of body weight intraperitoneally. The thymidine uptake in the DNA of the colon and of the small intestine were studied as an index of cell proliferation. A circadian rhythm in [³H]-thymiduie uptake in the colon was found and validated by cosinor analysis. This rhythm was similar in acrophase and amplitude in the animals kept on LD 8:16 and LD 12:12. Also in the mice on LD 16:8, there was a statistically significant circadian rhythm of ('HJ-thymidtne uptake in the DNA of the colon during all four seasons. The acrophases of this rhythm, however, varied widely suggesting free running. A circadian rhythm of pHJ-thymidine uptake in small intestine was less consistent. In animals on all three lighting regimens, however, a circannual variation of f'HJ-thymidine uptake in DNA in colon and small intestine was found with the highest uptake during summer. This study indicates that a lighting regimen of LD 16:8 does not reliably synchronize the circadian rhythm of [³H]-thymidine uptake in the colon. It further shows a circannual rhythm of this function in the colon and in the small intestine which persists under three lighting regimens (LD 8:16, 12:12 and 16:8) maintained for three to four weeks prior to being killed.     

Chronobiological study of several enzymes of lysosomal origin in human plasma

The possible occurrence of circadian and circannual rhythms in the plasma concentrations of the following enzymes of lysosomal origin was assessed: beta-D-N-acetylglucosaminidase (EC 3.2.1.30) beta-D-glucuronidase (EC 3.2.1.31), beta-D-glucosidase (EC 3.2.1.21), beta-D-galactosidase (EC 3.2.1.22), alpha-D-galactosidase (EC 3.2.1.23), alpha-L-fucosidase (EC 3.2.1) and alpha-D-mannosidase (EC 3.2.1.24). The circadian rhythm was studied in 16 women (aged: 17-24 years) and 13 men (age: 23 years) volunteers; the circannual rhythm, in 10 women and 8 men (age: 20-25 years). The circadian rhythm was detected in all the tested enzymes of women, and only in alpha-D-galactosidase, beta-D-glucosidase, alpha-D-mannosidase and beta-D-acetylglucosaminidase of men. A statistically significant difference between genders in the circadian rhythm was exhibited by beta-D-galactosidase (MESOR; amplitude) beta-D-glucosidase (MESOR; amplitude; acrophase) beta-D-N-acetylglucosaminidase, beta-D-glucuronidase and alpha-D-galactosidase (MESOR) and alpha-L-fucosidase (amplitude, acrophase). A circannual rhythm was detected in all the tested enzymes with the exception of beta-D-glucuronidase and beta-D-N-acetylglucosaminidase; no statistically significant difference between genders was detected. The group rhythms of some of the enzymes (alpha-D-galactosidase, beta-D-glucosidase, beta-D-galactosidase) showed similar values of both circadian and circannual acrophases, suggesting that they may subjected as a group to the same chronobiological coordination, possibly mediated by hormones. The chronobiological rhythms of lysosomal enzymes were different from those of lactate dehydrogenase and alpha 1-antitrypsin, indicating that these rhythms are not merely reflecting fluctuations of the water content of plasma. No in-phase relationship was observed between the circadian and circannual rhythms of plasma cortisol and those of the tested lysosomal enzymes, excluding a direct chronobiological and possibly functional relationship between this hormone and lysosomal enzymes.     

Circadian rhythmicity in leukocytes immune responses in the freshwater snake,Natrix piscator

Circadian rhythm is observed in most of the physiological functions including immune response. The use of animal models other than mammals is useful in understanding how the vertebrate circadian system is organized and how this biological clock has changed throughout the vertebrate evolution. The present study was aimed to examine the circadian variability in the innate immune responses of leukocytes in the freshwater snake, Natrix piscator. Leukocytes were isolated and processed for total and differential leukocyte count, leukocyte phagocytosis, NBT reduction, nitrite production, and lymphocyte proliferation. Experiments were conducted for seven time points at 24, 4, 8, 12, 16, 20, and 24 h in three seasons – summer, winter, and spring. Cosinor analysis revealed that among leukocytes, only lymphocyte count showed circadian variation in summer. Percent phagocytosis and phagocytic index had significant rhythm of 24 h in winter and summer season, respectively. The acrophase of NBT reduction and nitrite release were coming during the evening hours in summer and during morning hours in winter and had circadian rhythmicity. A significant phase shift in nitrite release was observed with a trend of delayed phase shift from winter to summer. Circadian rhythm was also observed in lymphocyte proliferation (basal and concanavalin A stimulated). It is evident from the present study that animals synchronize their immune activity according to the time of the day and season. Enhancement of immune function helps the individual cope with seasonal stressors that would otherwise jeopardize the survival of animal.     

Twenty-four-hour rhythm patterns of plasma melatonin in short-day and long-day breeders maintained under natural environmental conditions

ABSTRACT

Photoperiodic treatments have been of practical interest in controlling seasonal reproduction in sheep, goats and horses. Melatonin is the principal mediator of the environmental photoperiodic message. To investigate the intra- and inter-subject variability of melatonin 24 h rhythm, ten female Italian Saddle horses (8–10 yrs old, mean body weight 525 ± 30 kg), ten female Sarda breed sheep (2–3 yrs old, mean body weight 40.5 ± 2.8 kg) and ten female Sarda breed goats (3–4 yrs old, mean body weight 38.9 ± 4.1 kg), housed individually in a 4 × 4 m soundproof box equipped with 50 × 100 cm opening windows, were subjected to a natural photoperiod of the vernal equinox (sunrise 06:00 h; sunset 18:00 h). Blood samples were collected from each animal, every 3 h over a 48 h period starting at 00:00 h of day 1 and ending at 00:00 h of day 3. Plasma melatonin concentrations were determined by direct radioimmunoassay (MelatoninDirect RIA, Labor Diagnostika Nord GmbH, Nordhorn, Germany). The application of single cosinor method substantiated a circadian rhythm of melatonin with a nocturnal peak in all studied species. The application of two-way ANOVA on the rhythmic parameters indicated statistically significant differences between the three species in all of the cosinor analysis-derived parameters of MESOR, amplitude, acrophase and robustness of rhythm. Analyses of intra- and inter-subject variability indicate that organization of the melatonin 24 h rhythm is characterized by great accuracy of control within and between the individuals of a breed. In conclusion, features of the 24 h rhythm of melatonin among species; however, the 24 h rhythmicity of melatonin each species showed high stability within the various subjects and within the same subject. These findings must be taken into consideration when applying photoperiod and melatonin treatments for breeding purposes.