Circadian rhythm of airways caliber and its autonomic modulation

ABSTRACT

The autonomic nervous system (ANS) is one of the effector pathways for circadian variation of many physiological parameters. Autonomic tone and airways caliber have been reported to exhibit circadian variation in separate studies. A simultaneous investigation of heart rate variability (HRV) and airway caliber might ascertain how airway caliber is modulated by autonomic tone. This study was planned to identify the variations in airway caliber and autonomic function tone during a 24-hour span. A total of 56 healthy male subjects with almost similar daily routines were studied. Time domain, frequency domain and nonlinear analysis of R-R interval from 5 min electrocardiogram (ECG) was done seven times during the daytime wake span at 3-hour intervals starting at 05:00 h in the morning until 23:00 h in the night. Simultaneously peak expiratory flow rate (PEFR) was determined using a mini Wright’s peak flow meter. Rhythmometric analysis was done for PEFR and HRV parameters. Significant circadian variation in low frequency (LF) and high frequency (HF) variance was identified in this group of healthy subjects. The circadian rhythm of LF variance was characterized by a gradual increase and corresponding reciprocal change in HF variance from morning until night. The LF/HF ratio and SD2/SD1 ratio reflecting sympatho-vagal balance showed low to high values from morning to evening. The acrophase of the PEFR temporal pattern is similar to that of LF power and almost opposite in phase to that of HF power. PEFR is positively correlated with LF power. The circadian rhythm of airway caliber co-varies with cardiac autonomic tone. It appears that the temporal pattern of cardiac autonomic tone precedes in time that of airways caliber, thereby suggesting the latter operates under the modulatory effect of the 24-hour pattern in sympatho-vagal balance.     

Alertness and psychomotor performance levels of marine pilots on an irregular work roster

ABSTRACT

Fatigue is recognized as an important safety concern in the transportation industry. In this study, our goal was to investigate how circadian and sleep–wake dependent factors influence St-Lawrence River pilots’ sleep–wake cycle, alertness and psychomotor performance levels at work. A total of 18 male St-Lawrence River ship pilots were recruited to a 16–21-day field study. Pilots’ chronotype, sleepiness and insomnia levels were documented using standardized questionnaires. Their sleep–wake cycle was documented by a sleep–wake log and wrist-worn activity monitoring. Subjective alertness and objective psychomotor performances were assessed ~5×/day for each work and rest day. Ship transits were distributed throughout the 24-h day and lasted on average (± SEM) 5.93 ± 0.67 h. Main sleep periods occurred mainly at night, and objectively lasted 6.04 ± 1.02 h before work days. When going to bed at the end of work days, pilots subjectively reported sleeping 7.64 ± 1.64 h in the prior 24 h. Significant diurnal and wake-dependent effects were observed for subjective alertness and objective psychomotor performance, with minimum levels occurring between 09:00 and 10:00. Thus, despite their irregular work schedule, ship pilots presented, as a group, a diurnal variation of alertness and psychomotor performance indicative of a day-oriented circadian system. Important inter-individual differences were observed on psychomotor performance mesor and phase. In individuals, earlier phases in psychomotor performance were correlated with earlier chronotype. This study indicates that both circadian and homeostatic processes modulate alertness and psychomotor performance levels with worst levels reached when long shifts ended in the morning. This work has potential applications as it indicates fatigue countermeasures considering both processes are scientifically based.     

Diurnal variation and reliability of the urine lactate concentration after maximal exercise

The postexercise urine lactate concentration is a novel valid exercise biomarker, which has exhibited satisfactory reliability in the morning hours under controlled water intake. The aim of the present study was to investigate the diurnal variation of the postexercise urine lactate concentration and its reliability in the afternoon hours. Thirty-two healthy children (11 boys and 21 girls) and 23 adults (13 men and 10 women) participated in the study. All participants performed two identical sessions of eight 25 m bouts of maximal freestyle swimming executed every 2 min with passive recovery in between. These sessions were performed in the morning and afternoon and were separated by 3–4 days. Adults performed an additional afternoon session that was also separated by 3–4 days. All swimmers drank 500 mL of water before and another 500 mL after each test. Capillary blood and urine samples were collected before and after each test for lactate determination. Urine creatinine, urine density and body water content were also measured. The intraclass correlation coefficient was used as a reliability index between the morning and afternoon tests, as well as between the afternoon test and retest. Swimming performance and body water content exhibited excellent reliability in both children and adults. The postexercise blood lactate concentration did not show diurnal variation, showing a good reliability between the morning and afternoon tests, as well as high reliability between the afternoon test and retest. The postexercise urine density and lactate concentration were affected by time of day. However, when lactate was normalized to creatinine, it exhibited excellent reliability in children and good-to-high reliability in adults. The postexercise urine lactate concentration showed high reliability between the afternoon test and retest, independent of creatinine normalization. The postexercise blood and urine lactate concentrations were significantly correlated in all cases, attesting to the validity of urine lactate as an index of anaerobic metabolism. We conclude that urine lactate, after normalization to creatinine, could be used in training practice either in the morning or in the afternoon. Further research is needed to assess the applicability of this novel exercise biomarker.     

Circadian and seasonal variations of physiological and biochemical determinants of acute myocardial infarction

Almost all cardiovascular events occur according to a circadian rhythm with a greater frequency in the morning on waking and when resuming activity, the mechanism and precise triggering events for myocardial infarction (MI) are not yet fully known. Multiple biologic functions show a diurnal and/or seasonal variation that may contribute to adverse cardiac outomes. Exogenous factors may also modulates these variations. The MI peak usually occurs between 07:00 and 12:00 h. This timing corresponds to the concurrent increase in platelet aggregability, blood concentration of cortisol, catecholamines, angiotensin II, myocardial oxygen demand and coagulation activity, while fibrinolytic activity is decreased. In this review paper we will point out the biological rhythms of a number of functions involved in acute myocardial infarction e.g. blood pressure, hormonal determinants, cholesterol, among others.     

The effect of time of day on short-term maximal performance in obese and non-obese young men

The purpose of this study was to examine time-of-day effects on short-term performance in obese young men. In a balanced study design, 30 boys were divided into two groups: obese (OG; n = 15) and non-obese (CG; n = 15) groups. Both groups performed anaerobic tests of strength and power (squat-Jump, SJ; counter-movement jump, CMJ; 10-m sprint; and 30-m sprint) at 08:00 and 17:00 h on separate days. The results showed morning–evening differences in all variables in CG, with significant increases at 17:00 h (around the time of peak temperature) in comparison with 08:00 h (p < 0.01) for OG. However, such morning-evening differences were not found in the OG, except for the 30-m sprint. Obesity affects the diurnal variation on performances of 30-m sprint with a significant decrease in the evening for OG of 1.23% (p < 0.01). A correlation between anaerobic performance and oral temperature was observed only in the 30-m sprint test. In conclusion, the findings of the present study indicated that obesity is a problem that can affect circadian rhythms and performance in obese young men; therefore, preventing childhood obesity should become a top priority in efforts to improve performance and health.     

Mid- and long-term correlations of plasma metabolite concentrations measured by a targeted metabolomics approach

Introduction

Plasma metabolites measured by metabolomics techniques have been shown to be associated with chronic disease risk in epidemiological studies. However, in most prospective studies metabolomic profiles can only be obtained at a single time point and data on intra-individual variation in metabolite levels over time are sparse.

Objectives

Here, we evaluated the intra-individual variation in the concentrations of 177 metabolites over time using repeat blood samples of 104 adults (50% female).

Methods

Blood samples were obtained at a baseline visit between 1994 and 1998 and during two further examinations 14 and 15 years later. Plasma metabolite levels were quantified by tandem mass spectrometry with the MetaDisIDQ? p180 Kit. Intra-individual variation was assessed by Spearman’s correlation coefficients (ρ).

Results

Mid-term correlations over 1 year were good (ρ ≥ 0.7) for 5.1% and reasonable (ρ ≥ 0.4 < 0.7) for 61.0% of the metabolites, while long-term correlations over 15 years were good for 2.8% and reasonable for 27.1%. The strongest mid-term correlations between metabolite concentrations were observed for the acylcarnitine C₃–OH (0.72) and metabolites from the amino acid/biogenic amine groups, i.e. creatinine (0.83), proline (0.79), lysine (0.77), isoleucine (0.76), and ornithine (0.74). C₃–OH (0.78) as well as several amino acids/biogenic amines, i.e. ornithine (0.76), sarcosine (0.76), lysine (0.75), spermine (0.73), and glutamine (0.69) showed the strongest long-term correlations.

Conclusions

The biological reproducibility of plasma concentrations of a majority of metabolites occurs reasonable over 1 year. In contrast, concentrations of many metabolites seem to be affected by substantial intra-individual variation over 15 years.

     

Arachidonic acid metabolites and their circadian rhythm in patients with allergic bronchial asthma

The aim of this study was to investigate circadian variation in concentrations of arachidonic acid (AA) metabolites in relation to the circadian pattern in bronchial patency. Blood samples were obtained at 4-hr intervals from 2000 of 1 day until 1400 of the next from 12 diurnally active asthmatic and six diurnally active non-asthmatic patients. Bloods were analyzed for the prostanoids thromboxane A2 (measured as stable metabolite 6-keto-PGF1a), PGE2 and PGF2a. Airways patency was assessed by self-measurement of peak expiratory flow (PEF). In asthmatics, circadian variation was detected in PEF as well as PGE2 and TXB2. The circadian trough of the PEF rhythm closely coincided with the circadian peak of the PGE2 and TXB2 rhythms. In the controls, the PEF was not circadian rhythmic. Of the AA metabolites only 6-keto-PGF1a exhibited 24-hr bioperiodicity in the controls. The controls exhibited a significantly higher circadian mean of PEF (P less than 0.001), while the asthmatics had a lower 24-hr average PGE2 but greater mean TXB2/PGE2 ratio. The obstructive effect caused by the overall 24-hr deficiency of PGE2 in asthmatics is possibly amplified by the increased of TXB2 during the early morning hours. This dissociation of the temporal patterns in TXB2 and PGE2 levels over the 24 hr is discussed as a characteristic finding for asthmatics.     

The fetal circadian rhythm in pregnancies complicated by pregestational diabetes is altered by maternal glycemic control and the morning cortisol concentration

Circadian rhythmicity is fundamental to human physiology, and is present even during fetal life in normal pregnancies. The impact of maternal endocrine disease on the fetal circadian rhythm is not well understood. The present study aimed to determine the fetal circadian rhythm in pregnancies complicated by pregestational diabetes mellitus (PGDM), compare it with a low-risk reference population, and identify the effects of maternal glycemic control and morning cortisol concentrations. Long-term fetal electrocardiogram recordings were made in 40 women with PGDM at 28 and 36 weeks of gestation. Two recordings were made in 18 of the women (45.0%) and one recording was made in 22 (55.0%). The mean fetal heart rate (fHR) and the fHR variation (root mean square of squared differences) were extracted in 1-min epochs, and circadian rhythmicity was detected by cosinor analysis. The study cohort was divided based on HbA1c levels and morning cortisol concentrations. Statistically, significant circadian rhythms in the fHR and the fHR variation were found in 45 (100%) and 44 (95.7%) of the 45 acceptable PGDM recordings, respectively. The rhythms were similar to those of the reference population. However, there was no statistically significant population-mean rhythm in the fHR among PGDM pregnancies at 36 weeks, indicating an increased interindividual variation. The group with higher HbA1c levels (>6.0%) had no significant population-mean fHR rhythm at 28 or 36 weeks, and no significant fHR-variation rhythm at 36 weeks. Similarly, the group with a lower morning cortisol concentration (≤8.8 µg/dl) had no significant population-mean fHR-variation rhythm at 28 and 36 weeks. These findings indicate that individual fetal rhythmicity is present in pregnancies complicated by PGDM. However, suboptimal maternal glycemic control and a lower maternal morning cortisol concentration are associated with a less-well-synchronized circadian system of the fetus.     

A Preliminary Investigation into Individual Differences in the Orcadian Variation of Meal Tolerance: Effects on Mood and Hunger

A forced desynchrony methodology was used to assess postprandial blood glucose in 9 female volunteers during a 3-h period following a mixed meal presented at four times of day (08:00, 14:00, 20:00, 02:00). The influence of time of day on the postmeal glucose responses was evaluated by calculating the area under the curve, largest increase, time taken to reach peak, and fasting level. Circadian variations in meal tolerance were found for the area under the curve and largest increase, responses were greater (indicating poorer meal tolerance) in the evening than the morning. Fasting blood glucose exhibited diurnal variation although in the opposite direction to meal tolerance; levels were higher in the morning than the evening. Time taken to reach peak levels was not modulated by circadian rhythmicity. Estimates of the timing of poorest meal tolerance and the magnitude of this intolerance were computed for each subject. Individual differences in the magnitude of meal intolerance were found to influence hunger and self-reported calmness. Subjects with good tolerance had rhythms in both calmness and hunger, which were not found in those with poor tolerance. Subjects with good tolerance also tended to rate themselves as feeling more calm. These mood and hunger effects may result from differences in insulin resistance, which is hypothesized to underlie the circadian variations in meal tolerance. (Chronobiology International, 13(6), 435-447, 1996)     

Daily rhythms in activity and mRNA abundance of enzymes involved in glucose and lipid metabolism in liver of rainbow trout,Oncorhynchus mykiss. Influence of light and food availability

ABSTRACT

We have investigated the magnitude of diurnal variation in back squat and bench press performance using the MuscleLab force velocity transducer. Thirty resistance-trained males (mean ± SD: age 21.7 ± 1.4 years; body mass 80.5 ± 4.5 kg; height 1.79 ± 0.06 m) underwent two sessions at different times of day: morning (M, 07:30 h) and evening (E, 17:30 h). Each session included a period when rectal temperature (T_rec) was measured at rest, a 5-min standardized 150 W warm-up on a cycle ergometer, then defined programme of bench press (at 20, 40 and 60 kg) and back squat (at 30, 50 and 70 kg) exercises. A linear encoder was attached to an Olympic bar used for the exercises and average force (AF), peak velocity (PV) and time-to-peak velocity (tPV) were measured (MuscleLab software; MuscleLab Technology, Langesund, Norway) during the concentric phase of the movements. Values for T_rec at rest were higher in the evening compared to morning values (0.48°C, P < 0.0005). Daily variations were apparent for both bench press and back squat performance for AF (1.9 and 2.5%), PV (8.3 and 12.7%) and tPV (?16.6 and ?9.8%; where a negative number indicates a decrease in the variable from morning to evening). There was a main effect for load where AF and tPV increased and PV decreased from the lightest load to the heaviest for both bench press and back squat (47.1 and 80.2%; 31.7 and 57.7%; ?42.1 and ?73.9%; P < 0.0005 where a negative number indicates a decrease in the variable with increasing load). An interaction was found only for tPV, such that the tPV occurs earlier in the evening than the morning at the highest loads (60 and 70 kg) for both bench press and back squat, respectively (mean difference of 0.32 and 0.62 s). In summary, diurnal variation in back squat and bench press was shown; and the tPV in complex multi-joint movements occurs earlier during the concentric phase of exercise when back squat or bench press is performed in the evening compared to the morning. This difference can be detected using a low cost, portable and widely available commercial instrument and enables translation of past laboratory/tightly controlled experimental research in to main-stream coaching practice.     

Evidence of a diurnal rhythm in implicit reward learning

Many aspects of hedonic behavior, including self-administration of natural and drug rewards, as well as human positive affect, follow a diurnal cycle that peaks during the species-specific active period. This variation has been linked to circadian modulation of the mesolimbic dopamine system, and is hypothesized to serve an adaptive function by driving an organism to engage with the environment during times where the opportunity for obtaining rewards is high. However, relatively little is known about whether more complex facets of hedonic behavior – in particular, reward learning – follow the same diurnal cycle. The current study aimed to address this gap by examining evidence for diurnal variation in reward learning on a well-validated probabilistic reward learning task (PRT). PRT data from a large normative sample (N = 516) of non-clinical individuals, recruited across eight studies, were examined for the current study. The PRT uses an asymmetrical reinforcement ratio to induce a behavioral response bias, and reward learning was operationalized as the strength of this response bias across blocks of the task. Results revealed significant diurnal variation in reward learning, however in contrast to patterns previously observed in other aspects of hedonic behavior, reward learning was lowest in the middle of the day. Although a diurnal pattern was also observed on a measure of more general task performance (discriminability), this did not account for the variation observed in reward learning. Taken together, these findings point to a distinct diurnal pattern in reward learning that differs from that observed in other aspects of hedonic behavior. The results of this study have important implications for our understanding of clinical disorders characterized by both circadian and reward learning disturbances, and future research is needed to confirm whether this diurnal variation has a truly circadian origin.     

α1-Adrenoreceptor activity does not explain lower morning endothelial-dependent, flow-mediated dilation in humans

Early morning reduction in endothelium-dependent, flow-mediated dilation (FMD) may contribute to the high incidence of sudden cardiac death at this time of day. The mechanisms underpinning diurnal variation in FMD are unclear, but potentially relate to a circadian rhythm in sympathetic nerve activity. We hypothesized that blockade of α(1)-mediated sympathetic nerve activity would act to attenuate the diurnal variation in FMD. In a randomized and placebo-controlled design, we measured brachial artery FMD in 12 participants (mean age = 26 yr, SD = 3) at 0600 and 1600 after ingestion of an α(1)-blocker (prazosin, 1 mg/20 kg body mass) or placebo. Arterial diameter and shear rate were assessed using edge-detection software. Heart rate and blood pressure were also measured. Data were analyzed using linear mixed modeling. Following placebo, FMD was 8 ± 2% in the morning compared with 10 ± 3% in the afternoon (P = 0.04). Blockade with prazosin led to a slight but nonsignificant increase in morning FMD (P = 0.24) and a significant (P = 0.04) decrease in afternoon FMD, resulting in no diurnal variation (P = 0.20). Shear rate did not differ in the morning or afternoon under either condition (P > 0.23). Blood pressure was lower following prazosin compared with placebo (P < 0.02), an effect that was similar at both times of day (P > 0.34). Heart rate and norepinephrine levels were higher in the afternoon following prazosin. These data indicate that α(1)-adrenoreceptor activity does not explain lower morning endothelium-dependent FMD.     

Excretion of corticosteroid metabolites in urine and faeces of rats.

Stress enhances the production of corticosteroids by the adrenal cortex, resulting in the increased excretion of their metabolites in urine and faeces. An intraperitoneal injection of radioactive corticosterone was applied to adult, male Sprague-Dawley rats to monitor the route and delay of excreted metabolites in urine and faeces. Peak concentrations appeared in urine after 3.2 +/- 1.9 h and in faeces after 16.7 +/- 4.3 h. Altogether about 20% of the recovered metabolites were found in urine and about 80% in faeces. Using high-performance liquid chromatography (HPLC), several peaks of radioactive metabolites were found. Some metabolites were detected by enzyme immunoassay (EIA) using two different antibodies (corticosterone, 11beta-OH-aetiocholanolone). There was a marked diurnal variation with low levels of faecal corticosterone metabolites in the evening and higher values in the morning. This diurnal variation was influenced neither by the intraperitoneal injection of isotonic saline nor by ACTH. However, the administration of dexamethasone eliminated the morning peak for 2 days.     

The internal circadian clock increases hunger and appetite in the evening independent of food intake and other behaviors

Objective:

Despite the extended overnight fast, paradoxically, people are typically not ravenous in the morning and breakfast is typically the smallest meal of the day. We assessed whether this paradox could be explained by an endogenous circadian influence on appetite with a morning trough, while controlling for sleep/wake and fasting/feeding effects.

Design and Methods:

Twelve healthy non‐obese adults (six males; age, 20‐42 years) were studied throughout a 13‐day laboratory protocol that balanced all behaviors, including eucaloric meals and sleep periods, evenly across the endogenous circadian cycle. Participants rated their appetite and food preferences by visual analog scales.

Results:

There was a large endogenous circadian rhythm in hunger, with the trough in the biological morning (8 AM) and peak in the biological evening (8 PM; peak‐to‐trough amplitude = 17%; P = 0.004). Similarly‐phased significant endogenous circadian rhythms were present in appetites for sweet, salty and starchy foods, fruits, meats/poultry, food overall, and for estimates of how much food participants could eat (amplitudes 14‐25%; all P < 0.05).

Conclusions:

In people who sleep at night, the intrinsic circadian evening peak in appetite may promote larger meals before the fasting period necessitated by sleep, whereas the circadian morning trough would theoretically facilitate the extended overnight fast. Furthermore, the circadian decline in hunger across the night would theoretically counteract the fasting‐induced hunger increase that could otherwise disrupt sleep.     

Peripheral clock system circadian abnormalities in Cushing’s disease

ABSTRACT

In Cushing’s syndrome, the cortisol rhythm is impaired and can be associated with the disruption in the rhythmic expression of clock genes. In this study, we evaluated the expression of CLOCK, BMAL1, CRY1, CRY2, PER1, PER2, PER3 genes in peripheral blood leukocytes of healthy individuals (n = 13) and Cushing’s disease (CD) patients (n = 12). Participants underwent salivary cortisol measurement at 0900 h and 2300 h. Peripheral blood samples were obtained at 0900 h, 1300 h, 1700 h, and 2300 h for assessing clock gene expression by qPCR. Gene expression circadian variations were evaluated by the Cosinor method. In healthy controls, a circadian variation in the expression of CLOCK, BMAL1, CRY1, PER2, and PER3 was observed, whereas the expression of PER1 and CRY2 followed no specific pattern. The expression of PER2 and PER3 in healthy leukocytes presented a late afternoon acrophase, similarly to CLOCK, whereas CRY1 showed night acrophase, similarly to BMAL1. In CD patients, the circadian variation in the expression of clock genes was lost, along with the abolition of cortisol circadian rhythm. However, CRY2 exhibited a circadian variation with acrophase during the dark phase in patients. In conclusion, our data suggest that Cushing’s disease, which is characterized by hypercortisolism, is associated with abnormalities in the circadian pattern of clock genes. Higher expression of CRY2 at night outlines its putative role in the cortisol circadian rhythm disruption.     

Diurnal Variation in Vascular Function: Role of Sleep

Although vascular function is lower in the morning than afternoon, previous studies have not assessed the influence of prior sleep on this diurnal variation. The authors employed a semiconstant routine protocol to study the contribution of prior nocturnal sleep to the previously observed impairment in vascular function in the morning. Brachial artery vascular function was assessed using the flow-mediated dilation technique (FMD) in 9 healthy, physically active males (mean?±?SD: 27?±?9 yrs of age), at 08:00 and 16:00?h following, respectively, 3.29?±?.37 and 3.24?±?.57?h prior sleep estimated using actimetry. Heart rate and systolic and diastolic blood pressures were also measured. The data of the experimental sleep condition were compared with the data of the “normal” diurnal sleep condition, in which FMD measurements were obtained from 21 healthy individuals who slept only during the night, as usual, before the morning test session. The morning-afternoon difference in FMD was 1?±?4% in the experimental sleep condition compared with 3?±?4% in the normal sleep condition (p?=?.04). This difference was explained by FMD being 3?±?3% lower in afternoon following the prior experimental sleep (p?=?.01). These data suggest that FMD is more dependent on the influence of supine sleep than the endogenous circadian timekeeper, in agreement with our previous finding that diurnal variation in FMD is influenced by exercise. These findings also raise the possibility of a lower homeostatic “set point” for vascular function following a period of sleep and in the absence of perturbing hemodynamic fluctuation. (Author correspondence: h.jones1@ljmu.ac.uk)     

A pilot replication study of two PER3 single nucleotide polymorphisms as potential genetic markers for morning and evening earliness-lateness

Polymorphisms in genes of circadian system family seem to be of most importance for understanding of mechanisms underlying self-assessed individual variation in morning–evening preference. A review of earlier reported positive findings indicated that, at least, four polymorphisms in period circadian clock 3 (PER3) showed significant association with, at least, one of sub-constructs of a morningness–eveningness scale. However, similar to other candidate gene studies, these studies suffer from increased likelihood of false positive findings. We tried to replicate some of the most recently published positive results on associations of sub-traits of morningness–eveningness with two PER3 non-synonymous single nucleotide polymorphisms. DNA from buccal swabs was collected from healthy residents of three Russian cities, Moscow (N = 149) and Novosibirsk and Stavropol (N = 248). The tested hypotheses were formulated in accord with the earlier reported positive findings: the rare alleles might be linked to a higher score on (i) morning earliness–lateness scale (rs2640909, Moscow data-set) and (ii) both morning and evening earliness–lateness scales (rs228729, Novosibirsk and Stavropol datasets). The results provided support for the former hypothesis. These and earlier reported results highlighted several critical issues that remained to be addressed in future independent replications of positive findings on potential genetic markers for morning and evening earliness–lateness.     

Melatonin-induced phase and dose responses in a diurnal mammal,Funambulus pennantii

ABSTRACT

Melatonin, an essential pineal hormone, acts as a marker of the circadian clock that regulates biological rhythms in animals. The effects of exogenous melatonin on the circadian system of nocturnal rodents have been extensively studied; however, there is a paucity of studies on the phase-resetting characteristics of melatonin in diurnal rodents. We studied the phase shifting effects of exogenous melatonin as a single melatonin injection (1 mg/kg) at various phases of the circadian cycle on the circadian locomotor activity rhythm in the palm squirrel, Funambulus pennantii. A phase response curve (PRC) was constructed. Adult male squirrels (N = 10) were entrained to a 12:12 h light-dark cycle (LD) in a climate-controlled chronocubicle with food and water provided ad libitum. After stable entrainment, squirrels were transferred to constant dark condition (DD) for free-running. Following stable free run, animals were administered a single dose of melatonin (1 mg/kg in 2% ethanol-phosphate buffered saline (PBS) solution) or vehicle (2% ethanol-PBS solution) at circadian times (CTs) 3 h apart to evoke phase shifts. The phase shifts elicited at various CTs were plotted to generate the PRC. A dose response curve was generated using four doses (0.5, 1, 2 and 4 mg/kg) administered at the CT of maximum phase advance. Melatonin evoked maximum phase advances at CT0 (1.23 ± 0.28 h) and maximum phase delays at CT15 (0.31 ± 0.09 h). In the dose response experiment, maximal phase shifts were evoked with 1 mg/kg. In contrast, no significant shifts were observed in control groups. Our study demonstrates that the precise timing and appropriate dose of melatonin administration is essential to maximize the amelioration of circadian rhythm–related disorders in a diurnal model.     

The variability of arsenic in blood and urine of humans

BackgroundHumans are exposed to inorganic and organic arsenic. The total arsenic (As) concentration in urine is a commonly used biomarker of exposure. However, little is known about variability of As in biological fluids and the diurnal variation of As excretion.ObjectivesMain objectives were to assess the variability of As in urine, plasma (P-As), whole blood (B-As), and the blood cell fraction (C-As), and to assess diurnal variation of As excretion.MethodsSix urine samples were collected at fixed times during 24 h on two different days around one week apart among 29 men and 31 women. Blood samples were collected when the morning urine samples were delivered. The intra-class correlation coefficient (ICC) was calculated as the ratio of the between-individuals variance to the total observed variance.ResultsGeometric mean (GM) 24 h urinary excretions of As (U-As_24 h) were 41 and 39 µg/24 h on the two days of sampling. Concentrations of B-As, P-As and C-As were highly correlated with U-As_24 h and As in first void morning urine. No statistically significant differences were observed for the urinary As excretion rate between the different sampling times. A high ICC was observed for As in the cellular blood fraction (0.803), while ICC for first morning urine corrected for creatine was low (0.316).ConclusionsThe study suggests that C-As is the most reliable biomarker for use in exposure assessment of individual exposure. Morning urine samples have low reliability for such use. No apparent diurnal variation was observed in the urinary As excretion rate.     

INTERACTIONS OF CORTISOL,TESTOSTERONE, AND RESISTANCE TRAINING: INFLUENCE OF CIRCADIAN RHYTHMS

Diurnal variation of sports performance usually peaks in the late afternoon, coinciding with increased body temperature. This circadian pattern of performance may be explained by the effect of increased core temperature on peripheral mechanisms, as neural drive does not appear to exhibit nycthemeral variation. This typical diurnal regularity has been reported in a variety of physical activities spanning the energy systems, from Adenosine triphosphate-phosphocreatine (ATP-PC) to anaerobic and aerobic metabolism, and is evident across all muscle contractions (eccentric, isometric, concentric) in a large number of muscle groups. Increased nerve conduction velocity, joint suppleness, increased muscular blood flow, improvements of glycogenolysis and glycolysis, increased environmental temperature, and preferential meteorological conditions may all contribute to diurnal variation in physical performance. However, the diurnal variation in strength performance can be blunted by a repeated-morning resistance training protocol. Optimal adaptations to resistance training (muscle hypertrophy and strength increases) also seem to occur in the late afternoon, which is interesting, since cortisol and, particularly, testosterone (T) concentrations are higher in the morning. T has repeatedly been linked with resistance training adaptation, and higher concentrations appear preferential. This has been determined by suppression of endogenous production and exogenous supplementation. However, the cortisol (C)/T ratio may indicate the catabolic/anabolic environment of an organism due to their roles in protein degradation and protein synthesis, respectively. The morning elevated T level (seen as beneficial to achieve muscle hypertrophy) may be counteracted by the morning elevated C level and, therefore, protein degradation. Although T levels are higher in the morning, an increased resistance exercise–induced T response has been found in the late afternoon, suggesting greater responsiveness of the hypothalamo-pituitary-testicular axis then. Individual responsiveness has also been observed, with some participants experiencing greater hypertrophy and strength increases in response to strength protocols, whereas others respond preferentially to power, hypertrophy, or strength endurance protocols dependent on which protocol elicited the greatest T response. It appears that physical performance is dependent on a number of endogenous time-dependent factors, which may be masked or confounded by exogenous circadian factors. Strength performance without time-of-day–specific training seems to elicit the typical diurnal pattern, as does resistance training adaptations. The implications for this are (a) athletes are advised to coincide training times with performance times, and (b) individuals may experience greater hypertrophy and strength gains when resistance training protocols are designed dependent on individual T response. (Author correspondence: Lawrence.hayes@uws.ac.uk)