Orcadian Rhythm of Serotonin Binding in Rat Brain—I. Effect of the Light-Dark Cycle

Moving rapidly from a supine to a standing posture is a common daily activity, yet a significant physiological challenge. Syncope can result from the development of initial orthostatic hypotension (IOH) involving a transient fall in systolic/diastolic blood pressure (BP) of >40/20?mm Hg within the first 15 s, and/or a delayed orthostatic hypotension (DOH) involving a fall in systolic/diastolic BP of >20/10?mm Hg within 15?min of posture change. Although epidemiological data indicate a heightened syncope risk in the morning, little is known about the diurnal variation in the IOH and DOH mechanisms associated with postural change. The authors hypothesized that the onset of IOH and DOH occurs sooner, and the associated cardiorespiratory and cerebrovascular changes are more pronounced, in the early morning. At 06:00 and 16:00?h, 17 normotensive volunteers, aged 26?±?1 yrs (mean?±?SE), completed a protocol involving supine rest, an upright stand, and a 60° head-up tilt (HUT) during which continuous beat-to-beat measurements of middle cerebral artery velocity (MCAv), mean arterial BP (MAP), heart rate, and end-tidal Pco₂ (P_ETco₂) were obtained. Mean MCAv was ～12% lower at baseline in the morning (p?≤?.01) and during the HUT (p?<?.01), despite a morning elevation in P_ETco₂ by ～2.2?mm Hg (p?=?.01). The decline in MAP during initial standing (morning vs. afternoon: 50%?±?4% vs. 49%?±?3%) and HUT (39%?±?3% vs. 38%?±?3%) did not vary with time-of-day (p?>?.30). In conclusion, although there is a marked reduction in MCAv in the morning, there is an absence of diurnal variation in the onset of and associated physiological responses associated with IOH and DOH. These responses, at least in this population, are unlikely contributors to the diurnal variation in orthostatic tolerance. (Author correspondence: N.C.Lewis@2004.ljmu.ac.uk)     

MAXIMAL POWER,BUT NOT FATIGABILITY,IS GREATER DURING REPEATED SPRINTS PERFORMED IN THE AFTERNOON

The present study was designed to investigate if the suggested greater fatigability during repeated exercise in the afternoon, compared to the morning, represents a true time-of-day effect on fatigability or a consequence of a higher initial power. In a counterbalanced order, eight subjects performed a repeated-sprint test [10?×?(6 s of maximal cycling sprint?+?30 s of rest)] on three different occasions between: 08:00–10:00, 17:00–19:00, and 17:00-19:00?h controlled (17:00–19:00?h_cont, i.e., initial power controlled to be the same as the two first sprints of the 08:00–10:00?h trial). Power output was significantly (p?<?0.05) higher for sprints 1, 2, and 3 in the afternoon than in the morning (e.g., sprint 1: 23.3 ±1 versus 21.2 ±1 W·kg^?1), but power decrement for the 10 sprints was also higher in the afternoon. Based on the following observations, we conclude that this higher power decrement is a consequence of the higher initial power output in the afternoon. First, there was no difference in power during the final five sprints (e.g., 20.4 ±1 versus 19.7 ±1 W·kg^?1 for sprint 10 in the afternoon and morning, respectively). Second, the greater decrement in the afternoon was no longer present when participants were producing the same initial power output in the afternoon as in the morning. Third, electromyographic activity of the vastus lateralis decreased during the exercise (p?<?0.05), but without a time-of-day effect. (Author correspondence: sebastien.racinais@aspetar.com)     

The Effect of Time-of-Day and Sympathetic α1-Blockade on Orthostatic Tolerance

Tolerance time to a standardized orthostatic stressor is markedly reduced in normotensive individuals in the morning. However, the physiological mechanisms that underpin this phenomenon are unknown. The purpose of this study was to examine the role of α₁-adrenergic activity on orthostatic tolerance and associated cardiorespiratory and cerebrovascular responses, and to determine whether its endogenous modulation is important in the diurnal variation of orthostatic tolerance. In a four-trial, randomized placebo-controlled crossover experiment, 12 normotensive volunteers (aged 25?±?1 yrs; mean?±?SE) completed a 60° head-upward tilt (HUT; 15?min or until onset of presyncope) at 06:00 and 16:00?h, 90?min after the administration of either α₁-blockade (prazosin, 1?mg/20?kg body weight) or placebo. Continuous beat-to-beat measurements of middle cerebral blood flow velocity (transcranial Doppler), blood pressure (Finometer), heart rate, stroke volume, cardiac output, and end-tidal carbon dioxide were obtained. Independent of time-of-day, α₁-blockade markedly reduced the ability to tolerate a 15-min 60° HUT; tolerance time was 229% shorter compared with the placebo condition (p?≤?.0001). Moreover, a marked diurnal variation in orthostatic tolerance was evident following α₁-adrenergic blockade; e.g., tolerance time in the morning (176?±?30 s) was lower than in the afternoon (354?±?75 s; p?=?.04). These findings highlight an important role of α₁-sympathetic vasoconstrictor activity in acutely regulating blood pressure and offsetting syncope, especially in the early morning. (Author correspondence: Nia.Lewis@ubc.ca)     

DIURNAL VARIATION IN WINGATE TEST PERFORMANCES: INFLUENCE OF ACTIVE WARM-UP

The purpose of the present study was to examine the effects of active warm-up duration on the diurnal fluctuations in anaerobic performances. Twelve physical education students performed a medical stress test (progressive test up to exhaustion) and four Wingate tests (measurement of peak power [P_peak], mean power [P_mean], and fatigue index during an all-out 30 s cycling exercise). The tests were performed in separate sessions (minimum interval?=?36?h) in a balanced and randomized design at 08:00 and 18:00?h, either after a 5?min (5-AWU) or a 15?min active warm-up (15-AWU). AWU consisted of pedaling at 50% of the power output at the last stage of the stress exhausting test. Rectal temperature was collected throughout the sessions. A two-way ANOVA (warm-up?×?time of day) revealed a significant interaction for P_peak (F_(1.11)?=?6.48, p?<?0.05) and P_mean (F_(1.11)?=?5.84, p?<?0.05): the time-of-day effect was significant (p?<?0.001) in contrast with the effect of warm-up duration (p?>?0.05). P_peak and P_mean improved significantly from morning to afternoon after both 5-AWU and 15-AWU, but the effect of warm-up duration was significant in the morning only. Indeed, the values of P_peak or P_mean were the same after both warm-up protocols in the afternoon. For rectal temperature, there was no interaction between time-of-day and warm-up duration. Rectal temperature before and after both the warm-up protocols was higher in the afternoon, and the effect of warm-up duration on temperature was similar at 08:00 and 18:00?h. In conclusion, the interpretation of the results of the anaerobic performance tests should take into account time-of-day and warm-up procedures. Longer warm-up protocols are recommended in the morning to minimize the diurnal fluctuations of anaerobic performances. (Author correspondence: n_souissi@yahoo.fr)     

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)     

Chronotype Predicts Activity Patterns in the Neural Underpinnings of the Motor System During the Day

Neuroimaging is increasingly used to study the motor system in vivo. Despite many reports of time-of-day influences on motor function at the behavioral level, little is known about these influences on neural motor networks and their activations recorded in neuroimaging. Using functional magnetic resonance imaging (fMRI), the authors studied 15 healthy subjects (9 females; mean?±?SD age: 23?±?3 yrs) performing a self-paced finger-tapping task at different times of day (morning, midday, afternoon, and evening). Blood-oxygenation-level-dependent signal showed systematic differences across the day in task-related motor areas of the brain, specifically in the supplementary motor area, parietal cortex, and rolandic operculum (p_corr?<?.0125). The authors found that these time-of-day-dependent hemodynamic modulations are associated with chronotype and not with homeostatic sleep pressure. These results show that consideration of time-of-day for the analysis of fMRI studies is imperative. (Author correspondence: roenneberg@lmu.de)     

Effect of Morning School Schedule on Sleep and Anthropometric Variables in Adolescents: A Follow-Up Study

The aim of this study was to assess whether the shift from afternoon to morning classes reduces the duration of sleep and whether this reduction has any relation to body fat measurements. This is a follow-up study in which students (n = 379), 12.4 (SD?±?0.7) yrs old, were evaluated before and after the school schedule shift, with a 1-yr interval between the first and second data collections. Adolescents were divided into two groups: an afternoon-morning group (students who shifted from afternoon to morning classes) and an afternoon-afternoon group (students who remained in afternoon classes). The morning schedule of classes lasted from 07:30 and 12:00?h, and the afternoon schedule of classes lasted from 13:00 and 17:30?h. Self-reported bedtime, wake-up time, and time-in-bed were obtained. Body mass index, waist circumference, and body fat percentage were obtained by direct measures. The results showed a reduction of time-in-bed during weekdays for those students who changed to the morning session (p < .001). Analysis of covariance (ANCOVA) for repeated measures of anthropometric differences between afternoon-afternoon and afternoon-morning groups showed no effect of the school schedule change on weight gain. In conclusion, the time-in-bed reduction in the period analyzed cannot be considered to be a mediating factor to modifications in overweight anthropometric indicators. (Author correspondence: michelleb_fisio@yahoo.com.br)     

PARADOXICAL POST-EXERCISE RESPONSES OF ACYLATED GHRELIN AND LEPTIN DURING A SIMULATED NIGHT SHIFT

Approximately 10% of employees undertake night work, which is a significant predictor of weight gain, possibly because responses to activity and eating are altered at night. It is known that the appetite-related hormone, acylated ghrelin, is suppressed after an acute bout of exercise during the day, but no researcher has explored whether evening exercise alters acylated ghrelin and other appetite-related outcomes during a subsequent night shift. Six healthy men (mean?±?SD: age 30?±?8 yrs, body mass index 23.1?±?1.1?kg/m²) completed two crossover trials (control and exercise) in random order. Participants fasted from 10:00?h, consumed a test meal at 18:00?h, and then cycled at 50% peak oxygen uptake or rested between 19:00–20:00?h. Participants then completed light activities during a simulated night shift which ended at 05:00?h. Two small isocaloric meals were consumed at 22:00 and 02:00?h. Venous blood samples were drawn via cannulation at 1?h intervals between 19:00–05:00?h for the determination of acylated ghrelin, leptin, insulin, glucose, triglyceride, and non-esterified fatty acids concentrations. Perceived hunger and wrist actimetry were also recorded. During the simulated night shift, mean?±?SD acylated ghrelin concentration was 86.5?±?40.8 pg/ml following exercise compared with 71.7?±?37.7 pg/ml without prior exercise (p?=?0.015). Throughout the night shift, leptin concentration was 263?±?242 pg/ml following exercise compared with 187?±?221 pg/ml without prior exercise (p?=?0.017). Mean levels of insulin, triglyceride, non-esterified fatty acids, and wrist actimetry level were also higher during the night shift that followed exercise (p?<?0.05). These data indicate that prior exercise increases acylated ghrelin and leptin concentrations during a subsequent simulated night shift. These findings differ from the known effects of exercise on acylated ghrelin and leptin during the day, and therefore have implications for energy balance during night work. (Author correspondence: c.j.morris@2006.ljmu.ac.uk).     

Diurnal Variation in Heart Rate Variability before and after Maximal Exercise Testing

As heart-rate variability (HRV) is under evaluation in clinical applications, the authors sought to better define the interdependent impact of age, maximal exercise, and diurnal variation under physiologic conditions. The authors evaluated the diurnal changes in HRV 24-h pre- and post-maximal aerobic exercise testing to exhaustion in young (19–25 yrs, n?=?12) and middle-aged (40–55 yrs, n?=?12) adults. Subjects wore a portable 5-lead electrocardiogram holter for 48?h (24?h prior to and following a maximal aerobic capacity test). Time-, frequency-, time-frequency-, and scale-invariant-domain measures of HRV were computed from RR-interval data analyzed using a 5-min window size and a 2.5-min step size, resulting in a different set of outputs every 2.5?min. Results were averaged (mean?±?SE) over four prespecified time periods during the morning, afternoon, evening, and night on Day 1 and Day 2. Diurnal changes in HRV in young and middle-aged adults were compared using a two-way, repeated-measures analysis of variance (ANOVA). Young adults demonstrated higher HRV compared to middle-aged adults during periods of wakefulness and sleep prior to maximal exercise stress testing (i.e., high-frequency power during Day 1: young adults: morning 1862?±?496?ms², afternoon 1797?±?384?ms², evening 1908?±?431?ms², and night 3202?±?728?ms²; middle-aged adults: morning 341?±?53?ms², afternoon 405?±?68?ms², evening 469?±?80?ms², and night 836?±?136?ms²) (p < .05). Exercise resulted in reductions in HRV such that multiple measures of HRV were not significantly different between age groups during the afternoon and evening periods. All measures of HRV demonstrated between-group differences overnight on Day 2 (p < .05). Young adults are associated with higher baseline HRV during the daytime. Sleep increases variability equally and proportionally to daytime variability. Given the higher baseline awake HRV and equal rise in HRV during sleep, the change in HRV from sleep to morning with exercise is greater in younger subjects. These physiologic results have clinical significance in understanding the pathophysiology of altered variability in ill patients. (Author correspondence: aseely@ohri.ca)     

Mutual Binding Inhibition of Tetrodotoxin and Saxitoxin to Their Binding Protein from the Plasma of the Puffer Fish,Fugu pardalis

The mutual binding inhibition of tetrodotoxin and saxitoxin to their binding protein from the plasma of Fugu pardalis was investigated by HPLC. The values for the half inhibitory concentration of tetrodotoxin (1.6 μM) binding to this protein (1.2 μM) for saxitoxin, and of saxitoxin (0.47 μM) binding to that (0.30 μM) for tetrodotoxin were 0.35±0.057 μM and 81±16 μM (n=2), respectively.     

Regulatory Properties of Prephenate Dehydrogenase and Prephenate Dehydratase from Corynebacterium glutamicum

Regulatory properties of the enzymes in l-tyrosine and l-phenyalanine terminal pathway in Corynebacterium glutamicum were investigated. Prephenate dehydrogenase was partially feedback inhibited by l-tyrosine. Prephenate dehydratase was strongly inhibited by l-phenylalanine and l-tryptophan and 100% inhibition was attained at the concentrations of 5 × 10^?2mm and 10^?1mm, respectively. l-Tyrosine stimulated prephenate dehydratase activity (6-fold stimulation at 1 mm) and restored the enzyme activity inhibited by l-phenylalanine or l-tryptophan. These regulations seem to give the balanced synthesis of l-tyrosine and l-phenyl-alanine. Prephenate dehydratase from C. glutamicum was stimulated by l-methionine and l-leucine similarly to the enzyme in Bacillus subtilis and moreover by l-isoleucine and l-histidine. C. glutamicum mutant No. 66, an l-phenylalanine producer resistant to p-fluorophenyl-alanine, had a prephenate dehydratase completely resistant to the inhibition by l-phenylalanine and l-tryptophan.     

Time-of-Day Mediates the Influences of Extended Wake and Sleep Restriction on Simulated Driving

Although a nonlinear time-of-day and prior wake interaction on performance has been well documented, two recent studies have aimed to incorporate the influences of sleep restriction into this paradigm. Through the use of sleep-restricted forced desynchrony protocols, both studies reported a time-of-day?×?sleep restriction interaction, as well as a time-of-day?×?prior wake?×?sleep dose three-way interaction. The current study aimed to investigate these interactions on simulated driving performance, a more complex task with ecological validity for the problem of fatigued driving. The driving performance of 41 male participants (mean?±?SD: 22.8 ±2.2 yrs) was assessed on a 10-min simulated driving task with the standard deviation of lateral position (SDLAT) measured. Using a between-group design, participants were subjected to either a control condition of 9.33?h of sleep/18.66?h of wake, a moderate sleep-restriction (SR) condition of 7?h of sleep/21?h of wake, or a severe SR condition of 4.66?h of sleep/23.33?h of wake. In each condition, participants were tested at 2.5-h intervals after waking across 7?×?28-h d of forced desynchrony. Driving sessions occurred at nine doses of prior wake, within six divisions of the circadian cycle based on core body temperature (CBT). Mixed-models analyses of variance (ANOVAs) revealed significant main effects of time-of-day, prior wake, sleep debt, and sleep dose on SDLAT. Additionally, significant two-way interactions of time-of-day?×?prior wake and time-of-day?×?sleep debt, as well as significant three-way interactions of time-of-day?×?prior wake?×?sleep debt and time-of-day?×?sleep debt?×?sleep dose were observed. Although limitations such as the presence of practice effects and large standard errors are noted, the study concludes with three findings. The main effects demonstrate that extending wake, reducing sleep, and driving at poor times of day all significantly impair driving performance at an individual level. In addition to this, combining either extended wake or a sleep debt with the early morning hours greatly decreases driving performance. Finally, operating under the influence of a reduced sleep dose can greatly decrease performance at all times of the day. (Author correspondence: raymond.matthews@unisa.edu.au)     

TIME-OF-DAY EFFECTS ON FATIGUE DURING A SUSTAINED ANAEROBIC TEST IN WELL-TRAINED CYCLISTS

The aim of this study was to evaluate time-of-day effects on fatigue during a sustained anaerobic cycling exercise. Sixteen healthy male competitive cyclists were asked to perform a 60 s Wingate test against a braking load of 0.087?kg.kg body mass^?1 during two experimental sessions, which were set up either at 06:00 or 18:00?h in counterbalanced order. There was only one session per day with a recovery period of at least 36?h between the two sessions. Each subject was trained to perform the test. The body mass used to determine the braking load was that of the first test session for each subject and remained constant throughout the two test periods. During the test, peak power (PP), mean power during the first 30 s (MP30 s) and the full 60 s of the test (MP60 s), and fatigue (i.e., the decrease in power output values throughout the exercise) were analyzed. Results confirmed the existence of diurnal variation in anaerobic power output. PP, MP30 s, and MP60 s were significantly higher at 18:00 than 06:00?h, with gains equal to 8.2, 7.8, and 7.8%, respectively. Moreover, all the power output values recorded in the evening were higher than those recorded in the morning, indicating that fatigue induced by this exercise is not affected by time-of-day in male competitive cyclists. It is hypothesized that the freedom and complexity of pedalling could allow adaptations in movement patterns, as a function of time-of-day, in order to maintain higher performance in the evening. For practical considerations, the more complex the movements required to perform a sport, the more the time-of-day effect can be taken into account and adapted to by the trained athlete, particularly in cyclic sporting disciplines such as swimming, running, rowing, and kayaking. (Author correspondence: damien.davenne@unicaen.fr)     

Production of l-rhamnulose,a rare sugar,from l-rhamnose using commercial immobilized glucose isomerase

Abstract

A commercial immobilized d-glucose isomerase from Streptomyces murines (Sweetzyme) was used to produce l-rhamnulose from l-rhamnose in a packed-bed reactor. The optimal conditions for l-rhamnulose production from l-rhamnose were determined as pH 8.0, 60?°C, 300?g L^?1 l-rhamnose as a substrate, and 0.6?h^?1 dilution rate. The half-life of the immobilized enzyme at 60?°C was 809?h. Under the optimal conditions, the immobilized enzyme produced an average of 135?g L^?1 l-rhamnulose from 300?g L^?1 l-rhamnose after 16 days at pH 8.0, 60?°C, and 0.6?h^?1 dilution rate, with a productivity of 81?g/L/h and a conversion yield of 45% in a packed-bed reactor.     

Optimization of the 503 antigen induction strategy of Leishmania infantum chagasi expressed in Escherichia coli M15

Abstract

Leishmaniosis is a complex of diseases that can be fatal, if not given proper attention. Despite its relevance in the public health system, there is no vaccine capable of preventing the disease in humans so far and its treatment is expensive and aggressive to human health. The present study aims to optimize the induction parameters of the 503 Leishmania i. chagasi antigen expressed in recombinant Escherichia coli M15. The induction at different cell densities was evaluated in order to analyze the influence of the induction time on the yield of the protein of interest. In this segment, lactose and isopropyl-β-d-thiogalactopyranoside (IPTG) were used as inducer molecules, using various concentrations: 0.1?g/L, 1.0?g/L, and 10?g/L for lactose and 20?μM, 100?μM, 500?μM, and 1000?μM for IPTG. The results presented that the concentration of IPTG that obtained the higher antigen levels was that of 100?μM (0.087?g/L), a 10-fold lower concentration than was being previously used in this type of system and for lactose, it was 1?g/L (0.016?g/L). Thus, the induction with 100?μM allowed obtaining the antigen with a concentration 5.6 times higher than the lactose induction maximum concentration.     

Properties of Crystalline ω-Amino Acid : Pyruvate Aminotransferase of Pseudomonas sp. F–126

ω-Amino acid: pyruvate aminotransferase, purified to homogeneity and crystallized from a Pseudomonas sp. F–126, has a molecular weight of 172,000 or 167,000±3000 as determined by the gel-filtration or sedimentation equilibrium method, respectively. The enzyme catalyzes the transamination between various ω-amino acids or amines and pyruvate which is the exclusive amino acceptor. α-Amino acids except l-α-alanine are inert as amino donor. The Michaelis constants are 3.3 mm for β-alanine, 19 mm for 2-aminoethane sulfonate and 3.3 mm for pyruvate. The enzyme has a maximum activity in the pH range of 8.5~10.5. The enzyme is stable at pH 8.0~10.0 and at up to 65°C at pH 8.0. Carbonyl reagents strongly inhibit the enzyme activity. Pyridoxal 5′-phosphate and pyridoxamine 5′-phosphate reactivate the enzyme inactivated by carbonyl reagents. The inhibition constants were determined to be 0.73 mm for d-penicillamine and 0.58 mm for d-cycloserine. Thiol reagents, chelating agents and l-α-amino acids showed no effect on the enzyme activity.     

TIME-OF-DAY EFFECTS ON COGNITION IN PREADOLESCENTS: A TRAILS STUDY

Cognitive performance fluctuates during the day due to diurnal variations in alertness level. This study examined: (1) whether cognitive performance in school-aged children is affected by time-of-day; (2) which functional domains are particularly vulnerable to time-of-day effects; and (3) whether the effects are more pronounced for cognitively more demanding tasks or task conditions. Children, aged 10–12 yrs, were randomly assigned to a test session starting either at 08:30 (n?=?802), 10:00 (n?=?713), or 13:00?h (n?=?652). Speed and accuracy of information processing were evaluated by tasks that assess input-related cognitive processes (e.g., stimulus encoding), central cognitive processes (e.g., working memory, sustained attention), and output-related processes (e.g., response organization) using the Amsterdam Neuropsychological Tasks program. Time-of-day effects in children were identified in specific neurocognitive domains, such as visuospatial processing and working memory, but only under cognitively more demanding task conditions. Sustained attention showed a speed-accuracy tradeoff with increased slowness and lapses in the early morning, but with better feedback responsiveness and perceptual sensitivity than in the early afternoon. Furthermore, there was a significant interaction of time-on-task with time-of-day for tempo, with the afternoon group increasing in tempo with time-on-task, and the early-morning group first showing a slowing of tempo with time-on-task, followed at the end of the task by a speed increase towards the initial levels. To conclude, the authors found time-of-day effects in preadolescents, which were confined to cognitively more demanding tasks tapping input-related and central cognitive processes. (Author correspondence: kbheijden@fsw.leidenuniv.nl)     

Ring the Bell for Matins: Circadian Adaptation to Split Sleep by Cloistered Monks and Nuns

Cloistered monks and nuns adhere to a 10-century-old strict schedule with a common zeitgeber of a night split by a 2- to 3-h-long Office (Matins). The authors evaluated how the circadian core body temperature rhythm and sleep adapt in cloistered monks and nuns in two monasteries. Five monks and five nuns following the split-sleep night schedule for 5 to 46 yrs without interruption and 10 controls underwent interviews, sleep scales, and physical examination and produced a week-long sleep diary and actigraphy, plus 48-h recordings of core body temperature. The circadian rhythm of temperature was described by partial Fourier time-series analysis (with 12- and 24-h harmonics). The temperature peak and trough values and clock times did not differ between groups. However, the temperature rhythm was biphasic in monks and nuns, with an early decrease at 19:39?±?4:30?h (median?±?95% interval), plateau or rise of temperature at 22:35?±?00:23?h (while asleep) lasting 296?±?39?min, followed by a second decrease after the Matins Office, and a classical morning rise. Although they required alarm clocks to wake-up for Matins at midnight, the body temperature rise anticipated the nocturnal awakening by 85?±?15?min. Compared to the controls, the monks and nuns had an earlier sleep onset (20:05?±?00:59?h vs. 00:00?±?00:54?h, median?±?95% confidence interval, p?=?.0001) and offset (06:27?±?0:22?h, vs. 07:37?±?0:33?h, p?=?.0001), as well as a shorter sleep time (6.5?±?0.6 vs. 7.6?±?0.7?h, p?=?.05). They reported difficulties with sleep latency, sleep duration, and daytime function, and more frequent hypnagogic hallucinations. In contrast to their daytime silence, they experienced conversations (and occasionally prayers) in dreams. The biphasic temperature profile in monks and nuns suggests the human clock adapts to and even anticipates nocturnal awakenings. It resembles the biphasic sleep and rhythm of healthy volunteers transferred to a short (10-h) photoperiod and provides a living glance into the sleep pattern of medieval time. (Author correspondence: isabelle.arnulf@psl.aphp.fr)     

Metabolism of Aromatic Amino Acid in Microorganisms

Phenylalanine ammonia-lyase, which catalyzes the conversion of l-phenylalanine to trans-cinnamic acid and ammonia, has been partially purified from the cells of Rhodotorula. Some of the properties of this phenylalanine ammoyia-lyase were investigated. The enzyme was stable in phosphate buffer of pH over the range of 6.0 to 7.0 On heating, the enzyme was stable up to 50°C, but above 60°C, it was destroyed. The enzyme activity was strongly inhibited by p-chloromercuribenzoate at 10^?5 m and almost recovered by the addition of glutathione or mercaptoethanol at 10^?3 m. The present enzyme preparation of Rhodotorula also catalyzed the deamination of l-tyrosine to trans-p-coumaric acid. trans-p-Coumaric acid was isolated from the reaction mixture and identified by its absorption spectra. The rates of deamination showed optima at pH 9.0 and 9.5 for l-phenylalanine and l-tyrosine, respectively.     

High levels of expression of multiple enzymes in the Smirnoff-Wheeler pathway are important for high accumulation of ascorbic acid in acerola fruits

ABSTRACT

Acerola fruits contain abundant ascorbic acid (AsA). The gene expression levels of three upstream enzymes in the primary AsA biosynthesis pathway were correlated with AsA contents in the fruits of two acerola cultivars. Multiple overexpression of the enzymes increased AsA contents, suggesting their high expression is important for high AsA accumulation in acerola fruits and the breeding of AsA-rich plants.

Abbreviations: AsA: ascorbic acid; PMI: phosphomannose isomerase; PMM: phosphomannomutase; GMP: GDP-d-mannose pyrophosphorylase; GME: GDP-d-mannose 3?,5?-epimerase; GGP: GDP-l-galactose phosphorylase; GPP: l-galactose-1-phosphate phosphatase; GDH: l-galactose dehydrogenase; GLDH: l-galactono-1,4-lactone dehydrogenase