Journal of Biological Rhythms

Eating at night time, as is frequent in shift workers, may contribute to increased cardiovascular disease (CVD) risk through a disruption in usual lipid metabolism, resulting in repeated and sustained hyperlipidemia at night. This systematic review aimed to investigate the impact of eating a meal at night compared with the same meal eaten during the day on postprandial lipemia. Six databases were searched: CINAHL Plus, Cochrane Library, EMBASE, Ovid MEDLINE, Informit, and SCOPUS. Eligible studies were original research cross-over design with a minimum fasting period of 5 h before testing preceded by a standardized control meal; measured postprandial triacylglycerol (TAG) for 5 h or greater; had meal time between 0700 h and 1600 h for day time and between 2000 h and 0400 h for night time; and had within-study test meals (food or drink) that were identical in macronutrient composition and energy. Two authors independently completed eligibility and quality assessment using the American Dietetic Association Quality Criteria Checklist for Primary Research. After removing duplicates, 4,423 articles were screened, yielding 5 studies for qualitative synthesis. All studies identified at least one parameter of the postprandial TAG response that was different as a result of meal time (e.g., the total concentration or the time course kinetics). Two studies reported a greater total TAG concentration (area under curve) at night compared with day, and 3 studies found no difference. Four studies reported that the kinetics of the postprandial time course of TAGs was different at night compared with during the day. Inconsistent reporting in the primary studies was a limitation of the review. Night eating may negatively affect postprandial lipemia and this review shows there is a need to rigorously test this using standardized methods and analysis with larger sample sizes. This is critical for informing strategies to lower CVD risk for shift workers.

The circadian clock is an evolutionarily conserved mechanism that generates the rhythmic expression of downstream genes. The core circadian clock drives the expression of clock-controlled genes, which in turn play critical roles in carrying out many rhythmic physiological processes. Nevertheless, the molecular mechanisms by which clock output genes orchestrate rhythmic signals from the brain to peripheral tissues are largely unknown. Here we explored the role of one rhythmic gene, Achilles, in regulating the rhythmic transcriptome in the fly head. Achilles is a clock-controlled gene in Drosophila that encodes a putative RNA-binding protein. Achilles expression is found in neurons throughout the fly brain using fluorescence in situ hybridization (FISH), and legacy data suggest it is not expressed in core clock neurons. Together, these observations argue against a role for Achilles in regulating the core clock. To assess its impact on circadian mRNA rhythms, we performed RNA sequencing (RNAseq) to compare the rhythmic transcriptomes of control flies and those with diminished Achilles expression in all neurons. Consistent with previous studies, we observe dramatic upregulation of immune response genes upon knock-down of Achilles. Furthermore, many circadian mRNAs lose their rhythmicity in Achilles knock-down flies, suggesting that a subset of the rhythmic transcriptome is regulated either directly or indirectly by Achilles. These Achilles-mediated rhythms are observed in genes involved in immune function and in neuronal signaling, including Prosap, Nemy and Jhl-21. A comparison of RNAseq data from control flies reveals that only 42.7% of clock-controlled genes in the fly brain are rhythmic in both males and females. As mRNA rhythms of core clock genes are largely invariant between the sexes, this observation suggests that sex-specific mechanisms are an important, and heretofore under-appreciated, regulator of the rhythmic transcriptome.

Temperature compensation and period determination by casein kinase 1 (CK1) are conserved features of eukaryotic circadian rhythms, whereas the clock gene transcription factors that facilitate daily gene expression rhythms differ between phylogenetic kingdoms. Human red blood cells (RBCs) exhibit temperature-compensated circadian rhythms, which, because RBCs lack nuclei, must occur in the absence of a circadian transcription-translation feedback loop. We tested whether period determination and temperature compensation are dependent on CKs in RBCs. As with nucleated cell types, broad-spectrum kinase inhibition with staurosporine lengthened the period of the RBC clock at 37°C, with more specific inhibition of CK1 and CK2 also eliciting robust changes in circadian period. Strikingly, inhibition of CK1 abolished temperature compensation and increased the Q₁₀ for the period of oscillation in RBCs, similar to observations in nucleated cells. This indicates that CK1 activity is essential for circadian rhythms irrespective of the presence or absence of clock gene expression cycles.

The circadian system drives the temporal organization of body physiology in relation to the changing daily environment. Shift-work (SW) disrupts this temporal order and is associated with the loss of homeostasis and metabolic syndrome. In a rodent model of SW based on forced activity in the rest phase for 4 weeks, we describe the occurrence of circadian desynchrony, as well as metabolic and liver dysfunction. To provide better evidence for the impact of altered timing of activity, this study explored how long it takes to recover metabolic rhythms and behavior. Rats were submitted to experimental SW for 4 weeks and then were left to recover for one week. Daily locomotor activity, food intake patterns, serum glucose and triglycerides, and the expression levels of hepatic Pparα, Srebp-1c, Pepck, Bmal1 and Per2 were assessed during the recovery period and were compared with expected data according to a control condition. SW triggered the circadian desynchronization of all of the analyzed parameters. A difference in the time required for realignment was observed among parameters. Locomotor activity achieved the expected phase on day 2, whereas the nocturnal feeding pattern was restored on the sixth recovery day. Daily rhythms of plasma glucose and triglycerides and of Pparα, Pepck and Bmal1 expression in the liver resynchronized on the seventh day, whereas Srebp-1c and Per2 persisted arrhythmic for the entire recovery week. SW does not equally affect behavior and metabolic rhythms, leading to internal desynchrony during the recovery phase.

Circadian misalignment between sleep and behavioral/feeding rhythms is thought to lead to various health impairments in shift workers. Therefore, we investigated how shift work leads to genome-wide circadian dysregulation in hospital nurses. Female nurses from the University of Alabama at Birmingham (UAB) Hospital working night shift (n = 9; 29.6 ± 11.4 y) and day shift (n = 8; 34.9 ± 9.4 y) participated in a 9-day study measuring locomotor activity and core body temperature (CBT) continuously. Additionally, cortisol and melatonin were assayed and peripheral blood mononuclear cells (PBMCs) were harvested for RNA extraction every 3 h on a day off from work. We saw phase desynchrony of core body temperature, peak cortisol, and dim light melatonin onset in individual night-shift subjects compared with day-shift subjects. This variability was evident even though day- and night-shift nurses had similar sleep timing and scheduled meal times on days off. Surprisingly, the phase and rhythmicity of the expression of the clock gene, PER1, in PBMCs were similar for day-shift and night-shift subjects. Genome-wide microarray analysis of PBMCs from a subset of nurses revealed distinct gene expression patterns between night-shift and day-shift subjects. Enrichment analysis showed that day-shift subjects expressed pathways involved in generic transcription and regulation of signal transduction, whereas night-shift subjects expressed pathways such as RNA polymerase I promoter opening, the matrisome, and endocytosis. In addition, there was large variability in the number of rhythmic transcripts among subjects, regardless of shift type. Interestingly, the amplitude of the CBT rhythm appeared to be more consistent with the number of cycling transcripts for each of the 6 subjects than was melatonin rhythm. In summary, we show that shift-work patterns affect circadian alignment and gene expression in PBMCs.

The human circadian system is primarily regulated by the 24-h LD cycle incident on the retina, and nocturnal melatonin suppression is a primary outcome measure for characterizing the biological clock’s response to those light exposures. A limited amount of data related to the combined effects of light level, spectrum, and exposure duration on nocturnal melatonin suppression has impeded the development of circadian-effective lighting recommendations and light-treatment methods. The study’s primary goal was to measure nocturnal melatonin suppression for a wide range of light levels (40 to 1000 lux), 2 white light spectra (2700 K and 6500 K), and an extended range of nighttime light exposure durations (0.5 to 3.0 h). The study’s second purpose was to examine whether differences existed between adolescents’ and adults’ circadian sensitivity to these lighting characteristics. The third purpose was to provide an estimate of the absolute threshold for the impact of light on acute melatonin suppression. Eighteen adolescents (age range, 13 to 18 years) and 23 adults (age range, 24 to 55 years) participated in the study. Results showed significant main effects of light level, spectrum, and exposure duration on melatonin suppression. Moreover, the data also showed that the relative suppressing effect of light on melatonin diminishes with increasing exposure duration for both age groups and both spectra. The present results do not corroborate our hypothesis that adolescents exhibit greater circadian sensitivity to short-wavelength radiation compared with adults. As for threshold, it takes longer to observe significant melatonin suppression at lower CS levels than at higher CS levels. Dose-response curves (amount and duration) for both white-light spectra and both age groups can guide lighting recommendations when considering circadian-effective light in applications such as offices, schools, residences, and healthcare facilities.

The intrinsically photosensitive retinal ganglion cells are the main conduit of the light signal emanating from the retina to the biological clock located in the suprachiasmatic nuclei of the hypothalamus. Lighting manufacturers are developing white light sources that are devoid of wavelengths around 480 nm (“cyan gap”) to reduce their impact on the circadian system. The present study was designed to investigate whether exposure to a “cyan-gap,” 3000 K white light source, spectrally tuned to reduce radiant power between 475 and 495 nm (reducing stimulation of the melanopsin-containing photoreceptor), would suppress melatonin less than a conventional 3000 K light source. The study’s 2 phases employed a within-subjects experimental design involving the same 16 adult participants. In Phase 1, participants were exposed for 1 h to 3 experimental conditions over the course of 3 consecutive weeks: 1) dim light control (<5 lux at the eyes); 2) 800 lux at the eyes of a 3000 K light source; and 3) 800 lux at the eyes of a 3000 K, “cyan-gap” modified (3000 K mod) light source. The same protocol was repeated in Phase 2, but light levels were reduced to 400 lux at the eyes. As hypothesized, there were significant main effects of light level (F_1,12 = 9.1, p < 0.05, η_p² = 0.43) and exposure duration (F_1,12 = 47.7, p < 0.05, η_p² = 0.80) but there was no significant main effect of spectrum (F_1,12 = 0.16, p > 0.05, η_p² = 0.01). There were no significant interactions with spectrum. Contrary to our model predictions, our results showed that short-term exposures (≤ 1 h) to “cyan-gap” light sources suppressed melatonin similarly to conventional light sources of the same CCT and photopic illuminance at the eyes.

Social jetlag, a misalignment between sleep timing on the weekend and during the work week, is associated with depressive symptoms among adults across both sexes. A previous study found that later sleep timing was associated with depressive symptoms in women but not men. To date, however, no research has investigated whether the association between social jetlag and depression varies by sex among adolescents. The current study assessed self-reported sleep, depressive symptoms, and demographic information from 3058 adolescents (48% female, mean [SD] age 15.59 [0.77] years) from the age 15 wave of the Fragile Families and Child Wellbeing Study (FFCWS). Social jetlag was calculated as the absolute value of the midpoint of sleep on the weekend minus the midpoint of sleep during the school week. Depressive symptoms were measured through a modified 5-item version of the Center for Epidemiologic Studies Depression Scale (CES-D). We assessed whether the associations among sleep duration on school nights, social jetlag, and depressive symptoms were similar between male and female adolescents using multiple linear regression. In fully adjusted models, sex moderated the association between school night total sleep time and depressive symptoms (p < 0.001) and between social jetlag and depressive symptoms (p = 0.037). In females, but not in males, school night total sleep time was negatively associated with depressive symptoms (p < 0.001), whereas social jetlag (p < 0.001) was positively and independently associated with depressive symptoms. The results indicate the importance of regular sleep timing across the week and adequate sleep duration for the maintenance of optimal emotional health among female adolescents.

Cyanobacteria contain a circadian oscillator that can be reconstituted in vitro. In the reconstituted circadian oscillator, the phosphorylation state of KaiC oscillates with a circadian period, spending about 12 h in the phosphorylation phase and another 12 h in the dephosphorylation phase. Although some entrainment studies have been performed using the reconstituted oscillator, they were insufficient to fully explain entrainment mechanisms of the cyanobacterial circadian clock due to the lack of input pathway components in the in vitro oscillator reaction mixture. Here, we investigate how an input pathway component, CikA, affects the phosphorylation state of KaiC in vitro. In general, CikA affects the amplitude and period of the circadian oscillation of KaiC phosphorylation by competing with KaiA for the same binding site on KaiB. In the presence of CikA, KaiC switches from its dephosphorylation phase to its phosphorylation phase prematurely, due to an early release of KaiA from KaiB as a result of competitive binding between CikA and KaiA. This causes hyperphosphorylation of KaiC and lowers the amplitude of the circadian oscillation. The period of the KaiC phosphorylation oscillation is shortened by adding increased amounts of CikA. A constant period can be maintained as CikA is increased by proportionally decreasing the amount of KaiA. Our findings give insight into how to reconstitute the cyanobacterial circadian clock in vitro by the addition of an input pathway component, and explain how this affects circadian oscillations by directly interacting with the oscillator components.