Web-Based Chronobiological Analysis

In this paper we describe a computer application that allows users from all over the world to perform chronobiological analyses with popular Internet browsers. Our application is based on the client-server philosophy, and it has been designed so as to minimize the network resources and connections to be used. Presently, our application includes several well-known data analysis techniques, mainly, that known as the cosinor method and those related. All the problems concerning security restrictions are solved, so that the user can have a real interactive chronobiological application.     

Recording the Motor Activity of Aquatic Animals by Ultrasound

An ultrasonic technique based on the Doppler effect is presented to allow the registration of aquatic animal movements. The method was tested on crucians (Carassius carassius) in an aquarium as part of chronobiological investigations. Comparison with light barrier measurements shows that the Doppler technique is suitable for detecting typical circadian and seasonal changes in the motor activity of the fishes. The features of the ultrasonic method are discussed.     

SUPPLEMENTARY ADMINISTRATION OF ARTIFICIAL BRIGHT LIGHT AND MELATONIN AS POTENT TREATMENT FOR DISORGANIZED CIRCADIAN REST-ACTIVITY AND DYSFUNCTIONAL AUTONOMIC AND NEUROENDOCRINE SYSTEMS IN INSTITUTIONALIZED DEMENTED ELDERLY PERSONS

Increased daytime napping, early morning awakening, frequent nocturnal sleep interruptions, and lowered amplitude and phase advance of the circadian sleep-wake rhythm are characteristic features of sleep-waking and chronobiological changes associated with aging. Especially in elderly patients with dementia, severely fragmented sleep-waking patterns are observed frequently and are associated with disorganized circadian rhythm of various physiological functions. Functional and/or organic deterioration of the suprachiasmatic nucleus (SCN), decreased exposure to time cues such as insufficient social interaction and reduced environmental light, lowered sensitivity of sensory organs to time cues, and reduced ability of peripheral effector organs to express circadian rhythms may cause these chronobiological changes. In many cases of dementia, the usual treatments for insomnia do not work well, and the development of an effective therapy is an important concern for health care practitioner and researchers. Recent therapeutical trials of supplementary administration of artificial bright light and the pineal hormone melatonin, a potent synchronizer for mammalian circadian rhythm, have indicated that these treatments are useful tools for demented elderly insomniacs. Both bright light and melatonin simultaneously ameliorate disorganized thermoregulatory and neuroendocrine systems associated with disrupted sleep-waking times, suggesting a new, potent therapeutic means for insomnia in the demented elderly. Future studies should address the most effective therapeutic design and the most suitable types of symptoms for treatment and investigate the use of these tools in preventive applications in persons in early stages of dementia. (Chronobiology International, 17(3), 419–432, 2000)     

Functional Proteomics: A Promising Approach to Find Novel Components of the Circadian System

In the postgenome era, the analysis of entire subproteomes in correlation with their function has emerged due to high throughput technologies. Early approaches have been initiated to identify novel components of the circadian system. For example, in the marine dinoflagellate Lingulodinium polyedra, a chronobiological proteome assay was performed, which resulted in the identification of already known circadian expressed proteins as well as novel temporal controlled proteins involved in metabolic pathways. In the green alga Chlamydomonas reinhardtii, two circadian expressed proteins (a protein disulfide isomerase and a tetratricopeptide repeat protein) were identified by functional proteomics. Also, the first hints of temporal control within chloroplast proteins of Arabidopsis thaliana were identified by proteome analysis.     

Pentobarbital anaesthesia in the chronobiological studies

Administration of anaesthesia may influence specific aspects of in vivo animal experiments and is an especially important consideration for experiments conducted during the daytime. Although chronobiological studies investigating interactions between general anaesthesia and circadian rhythms are sparse, all suggest that general anaesthetic agents have a significant effect on circadian rhythms. To assess the suitability of pentobarbital anaesthesia in chronobiological studies, this study was performed using pentobarbital-anaesthetized (40 mg/kg, intraperitoneal) female Wistar rats after adaptation to a light–dark (LD) cycle (12 h:12 h). Heart rate, rectal temperature (RT), electrocardiographic parameters, autonomic nervous system activity, acid–base balance and plasma concentrations of Na⁺, K⁺, Ca²⁺ and Cl^- were evaluated for their dependence on the LD cycle. LD differences were found in heart rate and RT, measured before the administration of the anaesthetic agent. Pentobarbital anaesthesia eliminated LD differences in all electrophysiological parameters, parameters of heart rate variability (except RR intervals) and parameters of acid–base balance and ion concentrations. LD differences with borderline statistical significance were found only for Na⁺ levels, with a higher level in the light period (i.e. nonactive) of the rat regimen day. During pentobarbital anaesthesia, parasympathetic tone predominates and sympathetic activity is depressed. Spontaneously breathing rats under pentobarbital anaesthesia are in an asphyxic state independent of the LD cycle in in vivo experiments. Results of this study suggest that pentobarbital anaesthesia is not suitable for chronobiological studies. However, it is suitable for cardiovascular research that is done regardless of the circadian rhythmicity, because it does not cause significant changes in heart activity.     

Features of the circadian rhythm of temperature of the skin at children of 8-9 years and young men and girls

Problem of the present work was studying age features circadian rhythm of temperature of a skin with application of a method "Termochron iButton". Day-night rhythm of a skin temperature at two age-grades was investigated: boys and girls of 8-9 years and young men and girls of 20-22 years. For this purpose monitoring temperature during 48 hours with an interval of registration 10 minutes has been lead. Are revealed authentic chronobiological differences: mesor temperatures above at girls, than at boys, and at young men, above, than at girls. Amplitude of a circadian rhythm is above at boys and at girls. Researches chronobiological parameters in the different terms of day have shown, that an average level of temperature at night below at all examinees. The amplitude at adult people in the different terms of day does not differ, while at children it above in the night term.     

The Maximization of Mean Vector Length for the Identification of Unknown Periods in Biological Rhythms: An Extension to Continuous Variables

In a recent paper Rocchi and Ghiandoni proposed the maximization of mean vector length as a circular statistics tool for the preliminary identification of unknown and unforeseeable periods in biological rhythms, but its application was limited to unitless data. In this paper an extension of the method to the case of continuous variables collected at equidistant time intervals is proposed. The extended method is proposed as an exploratory tool in the analysis of chronobiological data. Two different examples of the application of this method (to edible white truffle harvesting data and to TSH level data) are also included and compared with the Fourier spectral analysis.     

Daily rhythms of body temperature and heat production of sibling Mastomys species from different ecosystems--the response to photoperiod manipulations

We compared body temperature (T_b) and metabolic rates, measured as oxygen consumption (VO₂), daily rhythms of two sibling species of the genus Mastomys. We also studied their responses to long day (16L: 8D, LD) and short day (8L: 16D, SD) photoperiod manipulations at a constant ambient temperature of 26 + 1 °C. We noted significant differences in T_b and VO₂ daily rhythm patterns, under SD and LD-acclimation between the sibling species. These differences explain adaptation to the climatic conditions that prevail in the different ecosystems where these species live. To the best of our knowledge, this is the first time that physiological differences between the two siblings are measured by using chronobiological methods.     

The Maximization of Mean Vector Length for the Identification of Unknown Periods in Biological Rhythms: An Extension to Continuous Variables

In a recent paper Rocchi and Ghiandoni proposed the maximization of mean vector length as a circular statistics tool for the preliminary identification of unknown and unforeseeable periods in biological rhythms, but its application was limited to unitless data. In this paper an extension of the method to the case of continuous variables collected at equidistant time intervals is proposed. The extended method is proposed as an exploratory tool in the analysis of chronobiological data. Two different examples of the application of this method (to edible white truffle harvesting data and to TSH level data) are also included and compared with the Fourier spectral analysis.     

Chronobiological effects on exercise performance and selected physiological responses

Previous studies investigating the impact of circadian rhythms on physiological variables during exercise have yielded conflicting results. The purpose of the present investigation was to examine maximal aerobic exercise performance, as well as the physiological and psychophysiological responses to exercise, at four different intervals (0800 hours, 1200 hours, 1600 hours, and 2000 hours) within the segment of the 24-h day in which strenuous physical activity is typically performed. Ten physically fit, but untrained, male university students served as subjects. The results revealed that exercise performance was unaffected by chronobiological effects. Similarly, oxygen uptake, minute ventilation and heart rate showed no time of day influences under pre-, submaximal, and maximal exercise conditions. Ratings of perceived exertion were unaffected by time of day effects during submaximal and maximal exercise. In contrast, rectal temperature exhibited a significant chronobiological rhythm under all three conditions. Under pre- and submaximal exercise conditions, significant time of day effects were noted for respiratory exchange ratio, while a significant rhythmicity of blood pressure was evident during maximal exercise. However, none of these physiological variables exhibited significant differential responses (percent change from pre-exercise values) to the exercise stimulus at any of the four time points selected for study. Conversely, resting plasma lactate levels and lactate responses to maximal exercise were found to be significantly sensitive to chronobiological influences. Absolute post-exercise plasma norepinephrine values, and norepinephrine responses to exercise (percent change from pre-exercise values), also fluctuated significantly among the time points studied. In summary, these data suggest that aerobic exercise performance does not vary during the time frame within which exercise is normally conducted, despite the fact that some important physiological responses to exercise do fluctuate within that time period. Accepted: 18 August 1997     

Changes in human blood pressure with season,age and solar cycles: A 26-year record

The record was studied of a 71-year-old, diurnally active (0700–2200 hours) male psychiatrist (G.N.) who self-measured systolic and diastolic blood pressure (BP_S and BP_D) mostly but not exclusively on Sunday mornings, from 1969 to 1994. A large about-yearly change was revealed which increased with age and was accompanied by a decreasing trend in the yearly rhythm-adjusted mean (MESOR;P<0.01). According to conventional criteria that specify only upper limits of acceptability, G.N. was hypertensive in summer and normotensive in other seasons. Since changes in both MESOR and circannual amplitude occurred, a systematic surveillance of BP is the chronobiological recommendation.     

A chronobiological study on the relation between heart rate and QT interval duration in newborn infants

The relation between heart rate and QT interval is the result of the autonomic nervous system control on cardiac function in healthy adults; accordingly, chronobiological studies have shown that adult subjects have circadian rhythms of heart rate (expressed as R-R interval) and QT interval in phase. We have employed chronobiological methods to study heart rate and QT interval relation in 10 newborn infants, who are known to have an immature cardiac control. Findings from this study indicate that not all the newborns show circadian rhythms of heart rate and QT interval and that when both rhythms are present they do not correlate like in the adults. Likely, this lack of relationship between heart rate and QT interval in newborns is due to different maturational stages of the newborns studied. As a practical implication, in newborn infants, mathematical correction of QT interval by heart rate is not a reliable method.     

A Circular Statistics Tool for the Preliminary Identification of Unknown Periods in Biological Rhythms: An Application

A tool for the preliminary identification of unknown and unforeseeable periods in biological rhythms is presented. This method, called 'maximization of mean vector length', is based on circular statistics assumptions. It can be used effectively as an exploratory tool in the analysis of chronobiological data, while it carries a high risk of type I statistical error when used as a confirmative method. However, a correction term is provided in order to reduce this risk. An example of the application of this method to a set of spontaneous delivery data is also included.     

Small shifts in diurnal rhythms are associated with an increase in suicide: The effect of daylight saving

Sleep and Biological Rhythms - Large disruptions of chronobiological rhythms are documented as destabilizing individuals with bipolar disorder; however, the impact of small phase altering events is...     

Heart Rate Orcadian Rhythm as a Biological Marker of Desynchronization in Major Depression: A Methodological and Preliminary Report

Heart rate (HR) was continuously monitored during successive 24-hr periods in 19 healthy subjects and 26 major depressed patients (DSM III-R). Recordings were performed after a 2-week wash-out period and the morningness or eveningness typology of each subject was determined. The chronobiological parameters and rhythm percentage (RP) were calculated by the single cosinor method from the smoothed HR curves of each subject. In normal subjects, HR follows a circadian rhythm (RP > 65%) with the lowest values at night. Morning type subjects have an earlier peak time (13:30) than evening type subjects (17:30). Major depressive patients were split into two groups; in the first one HR circadian rhythm was still present (RP > 63%) with a decrease in amplitude (24%) while in the second group, no circadian rhythm of HR could be detected (RP < 25%, decrease in amplitude > 70%). In the group of patients with a persisting HK circadian rhythm, no veritable phase advance was observed. Our results suggest that circadian HR rhythm, which can be easily studied with non-invasive methods, might represent a chronobiological marker of some depressions. Given the lag that exists between the rhythms of morning type and evening type subjects, our study also stresses the importance of taking into account this behavioural trait in chronobiological studies.     

Pharmfit-a Nonlinear Fitting Program for Pharmacology

A new program is presented for nonlinear fitting of data from pharmacological and chronobiological investigations. It contains functions for calculating data from ligand-binding studies and competition experiments, for the analysis of dose-response curves, for pharmacokinetic calculations, and for cosine analysis of harmonic and overlapping rhythms. In addition, it is possible to implement general equations by the user. The program allows data exchange with most spreadsheet, database, and graphics presentation programs, and accepts data from two widely used ambulatory 24-h blood-pressure monitoring systems. The fitting procedure uses the Marquardt-Levenberg algorithm. It calculates the weighted or the unweighted fit together with a great variety of statistics for estimation of goodness of fit. A graphics module permits graphical presentation of the fitted curve. Moreover, fitting of data to different models can be compared for the most likely fit and model discrimination statistics for improvement of further experiments are provided. To demonstrate the chronobiological application of the fitting program PHARMFIT, the analysis of telemetric heart rate data from rats is presented.     

PHARMFIT—a Nonlinear Fitting Program for Pharmacology

A new program is presented for nonlinear fitting of data from pharmacological and chronobiological investigations. It contains functions for calculating data from ligand-binding studies and competition experiments, for the analysis of dose-response curves, for pharmacokinetic calculations, and for cosine analysis of harmonic and overlapping rhythms. In addition, it is possible to implement general equations by the user. The program allows data exchange with most spreadsheet, database, and graphics presentation programs, and accepts data from two widely used ambulatory 24-h blood-pressure monitoring systems. The fitting procedure uses the Marquardt–Levenberg algorithm. It calculates the weighted or the unweighted fit together with a great variety of statistics for estimation of goodness of fit. A graphics module permits graphical presentation of the fitted curve. Moreover, fitting of data to different models can be compared for the most likely fit and model discrimination statistics for improvement of further experiments are provided. To demonstrate the chronobiological application of the fitting program PHARMFIT, the analysis of telemetric heart rate data from rats is presented.     

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.     

Lighting requirements of nocturnal primates in captivity: A chronobiological approach

From the results of chronobiological studies in 11 Aotus lemurinus (trivirgatus) griseimembra, 3 Galago garnettii, 5 Galago senegalensis, and 6 Microcebus murinus, inferences can be made on the most suitable lighting conditions for nocturnal primates kept in captivity. In each species studied light controls the daily periodic course of activity in a dual way. First, the light-dark (LD) cycle acts as the main Zeitgeber, entraining the endogenous circadian timing system (CTS) to the environmental periodicity. Second, the prevailing light intensity has a direct species-specific inhibiting or enhancing effect, masking the level of activity predetermined by the CTS. Marked inhibition of activity is caused especially by low light intensities during dark-time (D-time), which can also lead to drastically reduced food intake (e.g., in Aotus). Therefore, high-amplitude LD cycles should be applied which guarantee a stable external and internal synchronization of the various circadian rhythms of the organism, with a D-illumination intensity high enough to prevent light-induced impairments of the behavior of the animals. Up to now LD cycles of 12:12 h (100–1,000:0.5–0.01 lx; ≥5,000°K) have proved to be most suitable. Only in Microcebus should the D-illumination be reduced to about 10^?4 lx. Moreover, it must be considered that species with a photoperiodically controlled reproduction cycle require specific alterations of the L-time:D-time ratio.     

Twenty-four-Hour Rhythmic Variations of Histamine in Human Female Blood

We have previously observed that blood histamine levels undergo rhythmic 24 hour variations in male normal subjects. In the present study we examined whether similar 24 hour modifications were also present in female subjects. To this purpose, 60 healthy cycling women were enrolled for a chronobiological transverse study. To achieve experimental conditions as homogeneous as possible, the study was carried out on the first day of the menstrual flow. Histamine was determined in whole venous blood by a spectrophotofluorimetric method. The blood histamine levels showed significant rhythmic changes depending on the time of blood sampling, with 1 maximum and 1 minimum within a period of 24 h, with percentage rhythm 15.1, MESOR 0.15 μg/ml, amplitude 0.02 μg/ml and acrophase at 01:20 h. These results reveal the presence of 24 h cyclic variations of histamine levels in the female, as it occurred in male. However, the differences observed in the chronobiological rhythms of histamine between the two sexes may represent a peculiar feature of the first day of the menstrual cycle.