Reestablishment of ad libitum feeding following partial food deprivation: Impact on locomotor activity, visceral fat, food intake, and circadian glycemic curve.

Food deprivation has been associated with the development of metabolic pathologies. Few studies have explored the repercussions of a partial food deprivation following the reestablishment of an ad libitum diet. This study investigates the impact of a partial food deprivation (an 8-hour food intake restriction coupled with a 4-hour feeding window during the active phase) and the subsequent return to ad libitum feeding on the glycemic curve, food intake, and locomotor behavior. Wistar rats aged 45 days were subjected to 6 weeks of a partial food deprivation followed by 6 weeks of ad libitum feeding. Body weight, visceral fat, food intake, circadian glycemia, oral glucose tolerance, and locomotor activity were evaluated. It was found that the partial food deprivation resulted in the reduction of both the body weight and food intake; however, it increased visceral fat by 60%. Circadian glycemic values were altered at all intervals during the light phase, and glucose sensitivity improved at 60 minutes in the oral glucose tolerance test (OGTT). In the food-deprived group, the locomotor activity rhythm was reduced, with an observed delay in the peak of activity, reduction in total activity, and a decrease in the rhythmicity percentage. After the reestablishment of the ad libitum feeding, there was recovery of body weight, no difference in visceral fat, normalization of the food intake pattern, circadian glycemia, and oral glucose tolerance. Additionally, the return to ad libitum feeding restored locomotor activity, although the duration required for its complete recovery warrants further investigation. In conclusion, partial food deprivation induces physio-metabolic changes in rats, most of which are reversed after reestablishing ad libitum feeding.

Chronic jet lag reduces motivation and affects other mood-related behaviors in male mice.

Introduction: The circadian system regulates various physiological processes such as sleep-wake cycles, hormone secretion, metabolism, and the reaction to both natural and drug-based rewards. Chronic disruption of the circadian system caused by unsteady synchronization with light-dark (LD) schedules, such as advancing chronic jet lag (CJL), leads to adverse physiological effects and pathologies, and is linked with changes in mood and depressive behaviors in humans and rodent models. Methods: C57BL/6J male mice were subjected to circadian disruption through phase advances of 6 h every 2 days (CJL +6/2). Mice under 12:12-h LD cycle were used as controls. After 8 weeks under these conditions, a battery of behavioral tests was performed to assess if mood-related behaviors were affected. Results: Compared to controls under 24 h LD cycles, mice under CJL presented desynchronization of activity-rest rhythms that led to several behavioral impairments, including a decrease in motivation for food reward, and an increase in anxiety, anhedonia, and depressive-like behavior. Conclusion: Chronic circadian disruption, caused by an experimental CJL protocol, affects mood-related and reward-related behaviors in mice. Understanding the importance of the circadian system and its potential role for disruption due to CJL is important for maintaining good health and well-being.

Chronotype as a predictor of scholar performance in a full-time middle school / Cronotipo como preditor de desempenho em uma escola de ensino médio de tempo integral

The performance of day-to-day tasks, whether satisfactory or unsatisfactory, varies due to several environmental synchronizers, including the 24-hour light-dark cycle. For instance, human performance on physical and/or cognitive demanding activities reaches its peak during the day when the body temperature is at its circadian peak. Individual differences in the circadian peaks in temperature along with individuals' timing of sleep is referred to as chronotype. Here, we aimed to answer if (a) chronotypes affect the performance of students in a Brazilian full-time school with an early start time and if (b) there are differences in performance based on chronotype. We expected to find (a) a positive effect of the morning chronotype on the students' performance, particularly in subjects that take place in early morning; (b) while a negative effect of the evening chronotype in that same period. To address the effect of the chronotype on the students' scholar performance we build a Generalized Linear Mixed Model (GLMM). Results support the hypothesis that the students' performance is partially attributed to their chronotype. In particular, our findings shows that evening-type students are expected to have an increase of 0.038 (p ≤0.05) log counts on their performance in Portuguese classes compared to other chronotypes. Here we add evidence for the effect that individual chronotypes have on the students' performance in a Brazilian full-time middle school. Distinctive features of the studied Brazilian full-time middle school related to chronotypes are discussed.

O desempenho em tarefas no dia a dia, seja satisfatório ou insatisfatório, varia devido a diversos sincronizadores ambientais, incluindo o ciclo claro-escuro de 24 horas. Por exemplo, o desempenho humano em atividades físicas e/ou cognitivamente exigentes atinge seu pico durante o dia, quando a temperatura corporal está no pico circadiano. Diferenças individuais nos picos circadianos de temperatura, juntamente com o horário de sono dos indivíduos, são referidas como cronotipo. Neste trabalho o objetivo foi responder se a) os cronotipos afetam o desempenho dos estudantes em uma escola brasileira de período integral e se b) há diferenças no desempenho com base no cronotipo. Esperamos encontrar a) um efeito positivo do cronotipo matutino no desempenho dos estudantes, especialmente em disciplinas que ocorrem no período da manhã; b) enquanto um efeito negativo do cronotipo vespertino nesse mesmo período. Para endereçar o efeito do cronotipo no desempenho acadêmico dos estudantes, construímos um Modelo Linear Generalizado Misto (GLMM). Os resultados corroboraram a hipótese de que o desempenho dos estudantes é parcialmente atribuído ao seu cronotipo. Em particular, nossos resultados mostram que é esperado que os estudantes vespertinos tenham um aumento de 0,038 (p ≤ 0,05) pontos logarítmicos em seu desempenho em aulas de Língua Portuguesa em comparação com outros cronotipos. Assim, adicionamos evidências para o efeito que o cronotipo têm no desempenho dos estudantes em uma escola de ensino médio integral no Brasil. As características particulares da escola estudada e suas relações com os cronotipos foram discutidas.

Time-restricted eating and circadian rhythms: the biological clock is ticking.

Meal timing may be a critical modulator of health outcomes due to complex interactions between circadian biology, nutrition and human metabolism. As such, approaches that aim to align food consumption with endogenous circadian rhythms are emerging in recent years. Time-restricted eating (TRE) consists of limiting daily nutrient consumption to a period of 4 to 12 hours in order to extend the time spent in the fasted state. TRE can induce positive effects on the health of individuals with overweight and obesity, including sustained weight loss, improvement in sleep patterns, reduction in blood pressure and oxidative stress markers and increased insulin sensitivity. However, it is not fully clear whether positive effects of TRE are due to reduced energy intake, body weight or the truncation of the daily eating window. In addition, null effects of TRE in some populations and on some parameters of cardiometabolic health have been documented. Some evidence indicates that greater promotion of health via TRE may be achieved if the nutrient intake period occurs earlier in the day. Despite some promise of this dietary strategy, the effects of performing TRE at different times of the day on human cardiometabolic health, as well as the safety and efficacy of this dietary approach in individuals with cardiometabolic impairments, need to be evaluated in additional controlled and long-term studies.

Circadian modulation of motivation in mice.

Most living organisms have a circadian timing system adapted to optimize the daily rhythm of exposure to the environment. This circadian system modulates several behavioral and physiological processes, including the response to natural and drug rewards. Food is the most potent natural reward across species. Food-seeking is known to be mediated by dopaminergic and serotonergic transmission in cortico-limbic pathways. In the present work, we show evidence of a circadian modulation of motivation for food reward in young (4-months old) and aged (over 1.5 years old) C57BL/6 mice. Motivation was assayed through the progressive ratio (PR) schedule. Mice under a 12:12 light/dark (LD) cycle exhibited a diurnal rhythm in motivation, becoming more motivated during the night, coincident with their active phase. This rhythm was also evident under constant dark conditions, indicating the endogenous nature of this modulation. However, circadian arrhythmicity induced by chronic exposure to constant light conditions impaired the performance in the task causing low motivation levels. Furthermore, the day/night difference in motivation was also evident even without caloric restriction when using a palatable reward. All these results were found to be unaffected by aging. Taken together, our results indicate that motivation for food reward is regulated in a circadian manner, independent of the nutritional status and the nature of the reward, and that this rhythmic modulation is not affected by aging. These results may contribute to improve treatment related to psychiatric disorders or drugs of abuse, taking into account potential mechanisms of circadian modulation of motivational states.

Working Time Society consensus statements: Evidence-based effects of shift work on physical and mental health.

Potential effects of shift work on health are probably related to the misalignment between the light-dark cycle and the human activity-rest cycle. Light exposure at night mediates these effects, including social misalignment and leads to an inversion of activity and rest, which, in turn, is linked to changes in behaviours. This article reviews the epidemiological evidence on the association between shift work and health, and possible mechanisms underlying this association. First, evidence from findings of the meta-analyses and systematic reviews published in the last 10 yr is presented. In addition, it reports the larger single-occupation studies and recent large population-based studies of the general workforce. Koch's postulates were used to evaluate the evidence related to the development of disease as a result of exposure to shift work. Finally, we discussed limitations of the multiple pathways that link shift work with specific disorders and the methodological challenges facing shift work research. We concluded that the clearest indications of shift work being the cause of a disease are given when there is a substantial body of evidence from high quality field studies showing an association and there is good evidence from laboratory studies supporting a causal explanation of the link.

Night work effects on salivary cytokines TNF, IL-1ß and IL-6.

Shift work is unavoidable in modern societies, but at the same time disrupts biological rhythms and contributes to social distress and disturbance of sleep, health and well-being of shift workers. Shift work has been associated with some chronic diseases in which a chronic inflammatory condition may play a role. However, few studies investigating the association of cytokine and other inflammation markers with shift workers have been published in recent years. In this study we evaluated the effects of permanent night work on the production of tumor necrosis factor (TNF), interleukin-1ß (IL-1ß), IL-6 and melatonin in saliva. Another aim was to demonstrate the benefit of the use of salivary cytokines for studies in chronobiology, since it is an easy and non-invasive method that allows for sampling at several times. Thirty-eight healthy male workers, being 21 day workers and 17 night workers, agreed to participate in this study. Sleep was evaluated by actigraphy and activity protocols. Saliva was collected during three workdays approximately at the middle of the work shift and at bed and wake times of the main sleep episode. Saliva samples were then analyzed by enzyme-linked immunosorbent assay to measure TNF, IL-1ß, IL-6 and melatonin levels, and the results were submitted to non-parametric statistical analysis. The use of saliva instead of blood allowed for a greater number of samples from the same subjects, allowing identifying alterations in the daily production patterns of salivary cytokines TNF, IL-1ß and IL-6 that probably are linked to night work. Salivary TNF and IL-1ß levels were similar for day and night workers, with higher daily production after awakening, in the morning hours for day workers and in the afternoon for night workers. Both groups presented a significant daily variation pattern of these two cytokines. Day and night workers produced similar amounts of salivary IL-6. Nevertheless, the daily variation pattern observed among day workers, with a peak after awakening, was absent among night workers. Thus, in our study, night workers showed partially adjusted daily variation patterns for salivary TNF and IL-1ß, not seen for salivary IL-6. Results for salivary IL-6 could be better explained as a consequence of circadian disruption due to permanent night work. Our results suggest that the whole circadian system, including clocks and pineal gland, is involved in regulating cytokine profile in shift workers and that a coordinated production of these cytokines, important for an adequate inflammatory response, could be disturbed by shift work. The distinct effects that shift work may have on different cytokines could give some cues about the mechanisms involved in this association.

Don't just say no: Differential pathways and pharmacological responses to diverse nitric oxide donors.

Nitric oxide (NO) is a gaseous free radical molecule with a short half-life (∼1 s), which can gain or lose an electron into three interchangeable redox-dependent forms, the radical (NO), the nitrosonium cation (NO+), and nitroxyl anion (HNO). NO acts as an intra and extracellular signaling molecule regulating a wide range of functions in the cardiovascular, immune, and nervous system. NO donors are collectively known by their ability to release NOin vitro and in vivo, being proposed as therapeutic pharmacological tools for the treatment of several pathologies, such as cardiovascular disease. The highly reactive NO molecule is easily oxidized under physiological conditions to N-oxides, nitrate/nitrite and nitrogen dioxide. Different cellular responses are triggered depending on: 1) NO concentration [e.g., nanomolar for heme coordination in the allosteric site of guanylate cyclase (sGC) enzyme]; 2) the type of chemical bound to the nitrosated group (i.e., bound to nitrogen, N-nitro, or bound to sulphur atom, S-nitro) determining post-translational cysteine nitrosation; 3) the time-dependent availability of molecular targets. Classic NO donors are: organic nitrates (e.g., nitroglycerin, or glyceryl trinitrate, GTN; isosorbide mononitrate, ISMN), diazeniumdiolates having a diolate group [or NONOates, e.g., 2-(N,N-diethylamino)-diazenolate-2-oxide], S-nitrosothiols (e.g., S-nitroso glutathione, GSNO; S-nitroso-N-acetylpenicillamine, SNAP) or the organic salt sodium nitroprusside (SNP). In addition, nitroxyl (HNO) donors such as Piloty's acid and Angeli's salt can also be considered. The specific NO form released, as well as its differential reactivity to thiols, could act on different molecular targets and should be discussed in the context of: a) the type and amount of NO species determining the sensitivity of molecular targets (e.g., heme coordination, or S-nitrosation); b) the cellular redox state that could gate different effects. Experimental designs should take special care when choosing which NO donors to use, since different outcomes are to be expected. This article will comment recent findings regarding physiological responses involving NO species and their pharmacological modulation with donor drugs, especially in the context of the photic transduction pathways at the hypothalamic circadian clock.

The Times of Our Lives: Interaction Among Different Biological Periodicities.

Environmental cycles on Earth display different periodicities, including daily, tidal or annual time scales. Virtually all living organisms have developed temporal mechanisms to adapt to such changes in environmental conditions. These biological timing structures-ranging from microsecond to seasonal timing-may have intrinsic properties and even different clock machinery. However, interaction among these temporal systems may present evolutionary advantages, for example, when species are exposed to changing climatic conditions or different geographic locations. Here, we present and discuss a model that accounts for the circadian regulation of both ultradian (less than 24-h) and infradian (more than 24-h) cycles and for the interaction among the three time scales. We show two clear examples of such interaction: (i) between the circadian clock and the seasonal regulation of the Hypothalamic-Pituitary-Thyroid (HPT) axis; and (ii) between the circadian clock and the hypothalamic-nigrostriatal (HNS) ultradian modulation. This remarkable interplay among the otherwise considered isolated rhythms has been demonstrated to exist in diverse organisms, suggesting an adaptive advantage of multiple scales of biological timing.

Clock genes expression and locomotor activity are altered along the light-dark cycle in transgenic zebrafish overexpressing growth hormone.

In the present work it was demonstrated that transgenic Danio rerio overexpressing growth hormone (GH-transgenic) present either altered gene expression at a determined time point, or different expression pattern along the LD cycle, when compared with non-transgenic (NT) animals, in the positive and negative loops of the circadian system. Gene expression of clock paralogs was reduced in GH fish at the beginning of the dark phase, leading to diminished expression amplitude along the LD cycle. Furthermore, although no differences were observed between NT and GH animals for bmal1a and cry2b expression at each time point, only GH fish presented amplitude along the LD cycle. Also, the locomotor activity behavior was evaluated for both groups. GH-transgenic animals presented higher locomotor activity along the whole LD cycle when compared with NT animals. These data suggest that alterations in the gene expression patterns along the LD cycle of the positive and negative loops of the circadian system, could lead to altered locomotor activity behavior in GH-transgenic fish, and GH overexpression could be responsible for these alterations, either affecting the pathways involved in the expression of genes from the circadian system or altering the metabolism.

Circadian rhythm in melatonin release as a mechanism to reinforce the temporal organization of the circadian system in crayfish.

Melatonin (MEL) is a conserved molecule with respect to its synthesis pathway and functions. In crayfish, MEL content in eyestalks (Ey) increases at night under the photoperiod, and this indoleamine synchronizes the circadian rhythm of electroretinogram amplitude, which is expressed by retinas and controlled by the cerebroid ganglion (CG). The aim of this study was to determine whether MEL content in eyestalks and CG or circulating MEL in hemolymph (He) follows a circadian rhythm under a free-running condition; in addition, it was tested whether MEL might directly influence the spontaneous electrical activity of the CG. Crayfish were maintained under constant darkness and temperature, a condition suitable for studying the intrinsic properties of circadian systems. MEL was quantified in samples obtained from He, Ey, and CG by means of an enzyme-linked immunosorbent assay, and the effect of exogenous MEL on CG spontaneous activity was evaluated by electrophysiological recording. Variation of MEL content in He, Ey, and CG followed a circadian rhythm that peaked at the same circadian time (CT). In addition, a single dose of MEL injected into the crayfish at different CTs reduced the level of spontaneous electrical activity in the CG. Results suggest that the circadian increase in MEL content directly affects the CG, reducing its spontaneous electrical activity, and that MEL might act as a periodical signal to reinforce the organization of the circadian system in crayfish.

Involvement of dopamine signaling in the circadian modulation of interval timing.

Duration discrimination within the seconds-to-minutes range, known as interval timing, involves the interaction of cortico-striatal circuits via dopaminergic-glutamatergic pathways. Besides interval timing, most (if not all) organisms exhibit circadian rhythms in physiological, metabolic and behavioral functions with periods close to 24 h. We have previously reported that both circadian disruption and desynchronization impaired interval timing in mice. In this work we studied the involvement of dopamine (DA) signaling in the interaction between circadian and interval timing. We report that daily injections of levodopa improved timing performance in the peak-interval procedure in C57BL/6 mice with circadian disruptions, suggesting that a daily increase of DA is necessary for an accurate performance in the timing task. Moreover, striatal DA levels measured by reverse-phase high-pressure liquid chromatography indicated a daily rhythm under light/dark conditions. This daily variation was affected by inducing circadian disruption under constant light (LL). We also demonstrated a daily oscillation in tyrosine hydroxylase levels, DA turnover (3,4-dihydroxyphenylacetic acid/DA levels), and both mRNA and protein levels of the circadian component Period2 (Per2) in the striatum and substantia nigra, two brain areas relevant for interval timing. None of these oscillations persisted under LL conditions. We suggest that the lack of DA rhythmicity in the striatum under LL - probably regulated by Per2 - could be responsible for impaired performance in the timing task. Our findings add further support to the notion that circadian and interval timing share some common processes, interacting at the level of the dopaminergic system.

Minutes, days and years: molecular interactions among different scales of biological timing.

Biological clocks are genetically encoded oscillators that allow organisms to keep track of their environment. Among them, the circadian system is a highly conserved timing structure that regulates several physiological, metabolic and behavioural functions with periods close to 24 h. Time is also crucial for everyday activities that involve conscious time estimation. Timing behaviour in the second-to-minutes range, known as interval timing, involves the interaction of cortico-striatal circuits. In this review, we summarize current findings on the neurobiological basis of the circadian system, both at the genetic and behavioural level, and also focus on its interactions with interval timing and seasonal rhythms, in order to construct a multi-level biological clock.

Unwinding the molecular basis of interval and circadian timing.

Neural timing mechanisms range from the millisecond to diurnal, and possibly annual, frequencies. Two of the main processes under study are the interval timer (seconds-to-minute range) and the circadian clock. The molecular basis of these two mechanisms is the subject of intense research, as well as their possible relationship. This article summarizes data from studies investigating a possible interaction between interval and circadian timing and reviews the molecular basis of both mechanisms, including the discussion of the contribution from studies of genetically modified animal models. While there is currently no common neurochemical substrate for timing mechanisms in the brain, circadian modulation of interval timing suggests an interaction of different frequencies in cerebral temporal processes.