Chronopharmacology of methotrexate pharmacokinetics in childhood leukemia.

Survival has been shown to improve when maintenance therapy for acute lymphocytic leukemia in children is given at night rather during the day. We examined the possibility that diurnal variation in methotrexate pharmacokinetics may contribute to this improvement. In a crossover study, we determined the pharmacokinetics of intravenous methotrexate at 10:00 and 21:00 h in six children with standard or high-risk leukemia. During the study, children refrained from concomitant drugs (6-mercaptopurine and trimethoprim sulfamethoxazole). There was a significant fall in methotrexate plasma clearance at night (from 5.6 +/- 3 ml/min/kg to 4.7 +/- 2.3 ml/min/kg p < 0.05). Renal clearance of methotrexate tended to decrease at night and unbound renal clearance decreased significantly (from 17.5 +/- 1.7 ml/min/kg to 8.5 +/- 3.6 ml/min/kg p < 0.05). Creatinine clearance did not exhibit diurnal variation, when comparing two 12-h collections, but there was a significant decrease in the nonglomerular clearance of methotrexate (from 14.8 +/- 5.2 to 6 +/- 4 ml/min/kg). Because it is a weak organic acid, the tubular secretion of methotrexate depends on urinary pH. At night urinary pH is more acidic. This may result in more reabsorption and hence reduced renal clearance.     

Circadian Time-Dependent Kinetics of Theophylline and Its Modulation by Phenobarbital Pretreatment in Rats

The pharmacokinetics of theophylline (TPH, 10 mg/kg i.v.) were assessed in rats (natural light-dark span, June 9–10) after i.p. pretreatment with saline and 80 mg/kg phenobarbital (PB), respectively, for 3 consecutive days at either 07:00 h or at 19:000 h. Serum concentrations of TPH were assayed by high-performance liquid chromatography. No significant differences of the elimination rates of TPH could be found between the times of TPH administration (clearance: 1.17 ± 0.17 ml/kg/min at 07:00 h vs. 1.23 ± 0.17 ml/kg/min at 19:00 hours). PB premedication markedly accelerated TPH elimination. The increase in clearance values was more expressed when TPH was injected at 07:00 h than at 19:00 h (2.48 ± 0.67 vs. 2.06 ± 0.41 ml/kg/min, p < 0.01).     

Effects of water deprivation for 72 hours on the pharmacokinetics of a new carbapenem,DA-1131, in rats

Hormonal, physiological, and biochemical changes occurring in dehydrated patients could alter the pharmacokinetics of the drugs; therefore, the pharmacokinetics of DA-1131, a new carbapenem antibiotic, were investigated after 1-min intravenous administration of the drug at 50 mg/kg to control and 72-hr water-deprived rats. The impaired kidney and liver functions were observed in water-deprived rats on the basis of tissue microscopic examination. After intravenous infusion of the drug to water-deprived rats, the plasma concentrations of DA-1131 were higher and this resulted in a significantly greater total area under the plasma concentration-time curve from time zero to time infinity than those in control rats (4520 versus 3760 microg min/ml). This could be due to significantly slower total body clearance (CL) of DA-1131 in water-deprived rats (9.81 versus 14.1 ml/min/kg). The significantly slower CL of DA-1131 in water-deprived rats was due to significant decrease in both renal clearance (2.87 versus 5.13 ml/min/kg because of a significant decrease in 8-hr urinary excretion of unchanged DA-1131 [28.4 versus 39.9% of the intravenous dose] due to impaired kidney function) and nonrenal clearance (6.82 versus 8.66 ml/min/kg because of a significant decrease in the metabolism of DA-1131 in the kidney, as proved by the significant decrease in total renal DHP-I enzyme activity [1900 versus 2130 mU/each kidney]) in water-deprived rats. Water-deprivation did not alter the affinity of rat tissues to DA-1131.     

Chronopharmacokinetic Study of Gentamicin in Dogs

The purpose of this experiment was to determine whether the time of day of single intravenous doses of gentamicin affects the drug's pharmacokinetics in dogs maintained under a 12 h light (08:00 to 20:00 h), 12 h dark (20:00 to 08:00 h) cycle. Using a crossover design, 6 mixed‐breed male dogs received a single dose of 2 mg/kg of gentamicin at 8:00 or 20:00 h. Serial blood samples were collected and pharmacokinetic parameters were calculated following each timed dose. The concentration of the antibiotic was lower following the 08:00 h compared to the 20:00 h administration. When gentamicin was administered at 20:00 h, the initial concentration, mean residence time, and area under the disposition curve were significantly higher (p<0.05) and the apparent volume of distribution of the central compartment, apparent volume of distribution, apparent volume of distribution at steady‐state, and total body clearance (1.73±0.55 at 20:00 h versus 3.31±0.67 L/min/kg at 08:00 h) were significantly lower than for the 08:00 h administration (p<0.05). Our results show that the pharmacokinetics of gentamicin exhibits significant temporal variation when administered to dogs at different times of day.     

Renal metabolism of substance P in the dog.

Substance P is a potent vasodilatory, diuretic, and natriuretic agent. Since subcellular fractions of the kidney rapidly inactivate substance P in vitro, the present study was designed to examine this observation $\text{in}$$\text{vivo}$ in anesthetized dogs. Arterial, renal venous, and urinary levels of immunoreactive substance P were determined by radioimmunoassay and were found to be 117±11, 128±12 and 659±104 pg/ml, respectively. The urinary and fractional excretion of immunoreactive substance P were 122±22 pg/min and 6.6±2.0%, respectively. When substance P was infused intravenously, the arterial and renal venous plasma levels of immunoreactive material increased whereas the urinary levels did not change. Infusions of 50 ng/kg/min of substance P significantly decreased mean arterial pressure, urinary volume, creatinine clearance as well as the urinary excretion, clearance, and fractional excretion of immunoreactive substance P. During intrarenal infusion of ¹²⁵I-(8-Tyr) substance P, high levels of radioactive material were found in the urine and renal venous plasma which failed to migrate on thin layer chromatography with intact ¹²⁵I-(8-Tyr) substance P. Thus under these conditions, intact substance P was not released from the kidney into the urine or renal venous blood, but instead circulating substance P was rapidly and completely metabolized, probably by both vascular and tubular elements of the kidney.     

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).     

The low nanomolar levels of N<Superscript>G</Superscript>-monomethylarginine in serum and urine of patients with chronic renal insufficiency are not significantly different from control levels

Summary.  There are no reliable mean values of N^G-monomethylarginine (NMMA) in blood and urine of patients with renal insufficiency available in the literature. Therefore we investigate whether the NMMA levels are changed in blood and urinary excretion of nondialysed and dialysed patients with chronic renal insufficiency to evaluate whether NMMA may reach sufficiently increased concentrations in blood of the patients to exert toxic biological activity. In nondialysed as well as in dialysed patients we find no significant difference in serum concentration of NMMA between patients and controls. In nondialysed patients (all with a residual creatinine clearance lower than 15 ml/min), we find 94.5 ± 26.1 nM (mean ± SD) versus 94.6 ± 19.5 nM in controls. Similar levels are found in serum of haemodialysed patients (each with serum creatinine levels >700 μM): 83.0 ± 20.2 nM. The urinary excretion of NMMA in nondialysed patients is also not significantly different from the excretion of controls: 123 ± 110 in patients versus 157 ± 117 nmol/24 hrs in controls. Furthermore, the clearance of NMMA is much lower compared to the clearance of the dimethylarginine derivatives. Based on the literature, the low nanomolar levels of NMMA found in blood of patients with renal insufficiency do not support the statement that NMMA proper may act as a uremic toxin. Received April 3, 2002 Accepted October 7, 2002 Published online January 20, 2003 Acknowledgements We thank the University of Antwerp, FWO (Fonds voor Wetenschappelijk Onderzoek) (G.0027.97 and G.0394.00), the Born-Bunge Foundation, the OCMW Medical Research Foundation and Neurosearch Antwerp for the financial support. Authors' address: Bart Marescau, Laboratory of Neurochemistry and Behaviour, UIA T504, University of Antwerp, Universiteitsplein 1, B-2610 Wilrijk, Belgium, Fax: 00 32 3 8202618; E-mail: bartold.marescau@ua.ac.be     

Progress in Dodecafluoropentane Emulsion as a Neuroprotective Agent in a Rabbit Stroke Model

Dodecafluoropentane emulsion (DDFPe) in 250 nm nanodroplets seems to swell modestly to accept and carry large amounts of oxygen in the body at >29 °C. Small particle size allows oxygen delivery even into hypoxic tissue unreachable by erythrocytes. Using permanent cerebral embolic occlusion in rabbits, we assessed DDFPe dose response as a neuroprotectant at 7 and 24 h post-embolization without lysis of arterial obstructions and investigated blood pharmacokinetics. New Zealand White rabbits (N?=?56) received cerebral angiography and embolic spheres (diameter?=?700–900 μm) occluded middle and/or anterior cerebral arteries. Intravenous DDFPe dosing (2 %?w/v emulsion) began at 60 min and repeated every 90 min until sacrifice at 7 or 24 h post-embolization. Seven-hour groups: (1) control (embolized without treatment, N?=?6), and DDFPe treatment: (2) 0.1 ml/kg (N?=?7), (3) 0.3 ml/kg (N?=?9), (4) 0.6 ml/kg (N?=?8). Twenty-four-hour groups: (5) control (N?=?16), and DDFPe treatment: (6) 0.1 ml/kg (N?=?10). Infarcts as percent of total brain volume were determined using vital stains on brain sections. Other alert normal rabbits (N?=?8) received IV doses followed by rapid arterial blood sampling and GC-MS analysis. Percent infarct volume means significantly decreased for all DDFPe-treated groups compared with controls, p?=?<0.004 to <0.03. Blood DDFP (gas) half-life was 1.45?±?0.17 min with R?=?0.958. Mean blood clearance was 78.5?±?24.9 ml/min/kg (mean?±?SE). Intravenous DDFPe decreases ischemic stroke infarct volumes. Blood half-life values are very short. The much longer therapeutic effect, >90 min, suggests multiple compartments. Lowest effective dose and maximum effective therapy duration are not yet defined. Rapid development is warranted.     

Effects of cysteine on the pharmacokinetics and pharmacodynamics of intravenous and oral azosemide in rats with protein-calorie malnutrition

The effects of cysteine on the pharmacokinetics and pharmacodynamics of azosemide were investigated after intravenous (10 mg/kg) and oral (20 mg/kg) administration to male Sprague-Dawley rats fed on 23% protein diet (control rats), and 5% protein diet with (rats with PCMC) or without (rats with PCM) oral cysteine (250 mg/kg, twice daily for the fourth week) for 4 weeks. After intravenous administration to rats with PCMC, some pharmacokinetic parameters restored fully or more than the level of control rats; the time-averaged nonrenal clearance (2.70 versus 2.32 ml/min/kg) and apparent volume of distribution at steady state (160 versus 189 ml/kg) were comparable to those in control rats, however, the terminal half-life (34.7 versus 57.2 min) and mean residence time (73.3 versus 99.3 min) were significantly shorter, area under the plasma concentration-time curve from time zero to time infinity (AUC, 1930 versus 2680 microg min/ml) was significantly smaller, and time-averaged renal (2.24 versus 1.21 ml/min/kg) and total body (CL, 4.98 versus 3.65 ml/min/kg) clearances were significantly faster than those in control rats. This could be mainly due to significantly faster renal clearance and at least partly due to increased cytochrome P450 1A2 activity by cysteine supplementation. After intravenous administration to rats with PCMC, the total amount of 8-hr urinary excretion of unchanged azosemide was significantly greater (457 versus 305 microg/g body weight), however, the 8-hr urine output (15.3 versus 31.1 ml/g kidney) was not significantly different between control rats and rats with PCMC. This could be due to the fact that urine output seemed to reach an upper plateau from 10 mg/kg dose of azosemide in rats.     

Chronopharmacological Study of Cephalexin in Dogs

Recent studies have identified a 24 h rhythm in the expression and function of PEPT1 in rats, with significantly higher levels during the nighttime than daytime. Similarly, temporal variations have been described in glomerular filtration rate and renal blood flow, both being maximal during the activity phase and minimal during the rest phase in laboratory rodents. The aim of this study was to assess the hypothesis that the absorption of the first‐generation cephalosporin antibiotic cephalexin by dogs would be less and the elimination would be slower after evening (rest span) compared to morning (activity span) administration, and whether such administration‐time changes could impair the medication's predicted clinical efficacy. Six (3 male, 3 female; age 4.83±3.12 years) healthy beagle dogs were studied. Each dog received a single dose of 25 mg/kg of cephalexin monohydrate per os at 10∶00 and 22∶00 h, with a two‐week interval of time between the two clock‐time experiments. Plasma cephalexin concentrations were determined by microbiological assay. Cephalexin peak plasma concentration was significantly reduced to almost 77% of its value after the evening compared to morning (14.52±2.7 vs. 18.77±2.8 µg/mL) administration. The elimination half‐life was prolonged 1.5‐fold after the 22∶00 h compared to the 10∶00 h administration (2.69±0.9 vs. 1.79±0.2 h). The area under the curve and time to reach peak plasma concentration did not show significant administration‐time differences. The duration of time that cephalexin concentrations remained above the minimal inhibitory concentrations (MIC) for staphylococci susceptiblity (MIC=0.5 µg/mL) was>70% of each of the 12 h dosing intervals (i.e., 10∶00 and 22∶00 h). It can be concluded that cephalexin pharmacokinetics vary with time of day administration. The findings of this acute single‐dose study require confirmation by future steady‐state, multiple‐dose studies. If such studies are confirmatory, no administration‐time dose adjustment is required to ensure drug efficacy in dogs receiving an oral suspension of cephalexin in a dosage of 25 mg/kg at 12 h intervals.     

Day-Night Differences in the Kinetics and Dynamics of Insulin: Diabetic Versus Normal Yucatan Minipigs

Time-dependent variations in the pharmacokinetics and pharmacodynamics of insulin were studied at two times, 10:30 and 20:30 during the same day in normal and streptozotocin (STZ)-induced diabetic minipigs housed in L(06:00):D(18:00) using the intravenous insulin tolerance test. Following intravenous insulin (0.1 IU/kg) administration in normal minipigs, the time for the glucose level to reach nadir (t_nadir) was significantly longer in the evening than the morning [(a.m.: 30.4 (± 2.4) VS. P.M.: 38.5 (± 3.3) min] (p < 0.01), although maximum reduction of glucose level (nadir) in the morning and evening was not significantly different [a.m.: (-70 (± 2) VS. P.M.: -65 (± 5) %]. The rate of glucose decline (K_in) was significantly decreased in the evening [a.m.: 5.33 (± 0.71) VS. P.M.: 4.44 (± 0.54) %dBG/min] (p < 0.01), and the integrated glucose-lowering response (ABCB) was significantly higher in the evening than the morning [a.m.: 3.18 (± 0.38) VS. P.M.: 4.52 (± 0.30) (g/dl) * min] (p < 0.01). The area under the plasma insulin concentration curve was increased significantly in the evening [a.m.: 2.26 (± 0.174) VS. P.M.: 2.74 (± 0.18) (mU/ml) * min], while the morning plasma insulin half-life did not differ significantly from that in the evening [a.m.: 4.79 (± 0.36) VS. P.M.: 5.47 (± 0.47) min]. After induction of diabetes by intravenous STZ injections, minipigs became diabetic, baseline blood glucose was observed to increase from the range of 45–55 to 200–250 mg/dl while plasma insulin levels decreased from 7–12 to 3–5 uU/ml. In the STZ-induced diabetic minipigs, a higher dose (0.2 IU/kg) was used in the intravenous insulin tolerance test in an attempt to normalize the high glucose levels. Following intravenous     

Twelve-Hour Night Shifts of Healthcare Workers: A Risk to the Patients?

We assessed the impact of 12h fixed night shift (19:00–07:00h) work, followed by 36h of off-time, on the sleep–wake cycle, sleep duration, self-perceived sleep quality, and work-time alertness on a group composed of 5 registered and 15 practical nurses. Wrist actigraphy (Ambulatory Monitoring, Inc.), with data analysis by the Cole-Kripke algorithm, was applied to determine sleep/wake episodes and their duration. The sleep episodes were divided into six categories: sleep during the night shift (x¯=208.6; SD±90.6mins), sleep after the night shift (x¯=138.7; SD±79.6min), sleep during the first night after the night work (x¯=318.5; SD±134.6min), sleep before the night work (x¯=104.3; SD±44.1min), diurnal sleep during the rest day (x¯=70.5; SD±43.0min), and nocturnal sleep during the rest day (x¯=310.4; SD±188.9mins). A significant difference (p<.0001; T-test for dependent samples) was detected between the perceived quality of sleep of the three diurnal sleep categories compared to the three nocturnal sleep categories. Even thought the nurses slept (napped) during the night shift, their self-perceived alertness systematically decreased during it. Statistically significant differences were documented by one-way ANOVA (F=40.534 p<.0001) among the alertness measurements done during the night shift. In particular, there was significant difference in the level of perceived alertness (p<.0001) between the 7^th and 10^thh of the 12h night shift. These findings of decreased alertness during the terminal hours of the night shift are of concern, since they suggest risk of comprised patient care.     

The glomerular filtration rate,effective renal plasma flow,and renal blood flow in the adult male chacma baboon (Papio ursinus)

Abstract: The radionuclide determination of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) has been validated in man, but not in the primate. GFR, ERPF, and renal blood flow (RBF) were measured in a group of 12 adult male chacma baboons using radiopharmaceuticals. GFR was determined using ^99mtechnetium-labelled diethylenetriamine-pentacetic acid. ERPF was measured with ¹³¹iodine-labelled hippuran. RBF, body surface area, and kidney weights were calculated using standard formulae. GFR was 49 ± 11 ml/min and ERPF was 237.9 ± 54.2 ml/min. Calculated RBF was 430.7 ± 111.9 ml/min and 507.4 ± 138.4 ml/min/100g of renal tissue. The results are in agreement with those obtained using more laborious nonradioisotopic techniques such as para-aminohippurate (PAH) and creatinine clearance and could serve as baseline normal values in the adult male chacma baboon.     

The production and secretion of cortisol by baboon neonates.

The metabolic clearance rate (MCR), transfer constants (p), production (PR) and secretion (SR) rates of cortisol (F) andrortisone (E) were determined by the continuous infusion of {1,2³H}F and {4-¹⁴C}E into 5 neonates delivered prior to the parturition by cesarean section (164–179 days; term = 184 days) and into 5 newborns delivered spontaneously per vagina at term (166 – 187 days). In spontaneously delivered animals, MCR-E (X ± SE, 34.3 ± 7.0 1/day/kg was greater (P < 0.001) than MCR-F (14.9 ± 1.5 1/day/kg), pF to E (59.7 ± 8.9%) exceeded (P < 0.001) pE to F (17.8 ± 3.0%) and the percentage of F bound to serum proteins other than albumin (57.5 ± 6.2) was greater (P < 0.001) than that of E (27.0 ± 10.3) Although the serum E level (25.6 ± 3.6 μg/100 ml) was similar to that of F (33.5 ± 8.0 μg/100 ml), the PR-E (6.4 ± 1.3 μ/min/kg) was greater (P < 0.001) than PR-F (3.3 ± 0.5 μ/min/kg). Approximately eighty-five percent of the E and 65% of the F produced orginated by secretion.In animals delivered by cesarean section, the serum F concentration (32.4 ± 6.7 μ/100ml), pE to F (13.4 ± 2.8%) pF to E (80.0 ± 12.2%) PR-E (4.5 ± 0.2 μ/min/kg) and SR-E (3.9 ± 0.3 μ/min/kg) were not different from values for spontaneously delivered animals. Serum E levels (35.9 ± 1.6 μ/100 ml) were higher but MCR-F (6.7 ± 0.6 1/day/kg) and MCR-E (18.2 ± 0.41/day/kg) lower in neonates delivered by cesarean section. Serum Cortisol binding capacity (μg F bound/100 ml) was greater (P < 0.025) in neonates delivered by cesarean section (23.6 ± 2.6) than in spontaneously delivered animals (14.4 ± 2.0). As a result of these changes in F and E dynamics, PR-F (1.4 ± 0.3 μ/min/kg) and SR-F (0.9 ± 0.2 μ/min/kg) in neonates delivered by cesarean section were lower (P< 0.01) than corresponding values in spontaneously delivered newborns.It is concluded that the greater F secretion in animals delivered spontaneously than those delivered by cesarean section probably results from increased fetal adrenal 3β-hydroxysteroid dehydrogenase-isomerase activity, which as previously reported, occurs in late gestation in this species.     

A study of nephron function in normal tropical residents using the creatinine and lithium clearances

 The kidney bears the brunt of the demands of a tropical climate for water and electrolyte homeostasis. We hypothesised that a tropical climate may cause adaptive changes in the entire organism leading to altered renal function in our subjects. Hence renal function data for residents of a temperate climate may not be applic- able to tropical residents. We therefore sought to elucidate renal function in subjects residing in a tropical climate. We used lithium clearance, C _Li, a non-invasive tool for assessing proximal tubular function in humans, and endogenous creatinine clearance, C _Cr, to estimate proximal tubular function and glomerular function, respectively, in our subjects. We did this in order to establish whether or not nephron function in our subjects differs from that for residents of a temperate climate. Nineteen male and 12 female Ghanaian subjects aged between 15 and 48 years were studied. The estimated G _Cr was 117.3±6.6 ml/min for male subjects and 97±6.4 ml/min for female subjects. C _Li was 20.3±1.6 ml/min for male and 19.1±0.4 ml/min for female subjects, respectively. The estimated absolute reabsorption rate of fluid of proximal tubules was 97.0±6.0 ml/min for males and 78.1±6.0 ml/min for females. The percentage proximal fluid reabsorption for male and female subjects was 81.2±1.4 and 79.5±1.6, respectively. The differences between male and female values (mean±SEM) were not statistically significant. The data suggest that the proximal tubule in residents of a tropical climate may reabsorb more fluid compared to that in residents of a temperate climate. Our values for proximal tubular reabsorption are higher than those reported for residents of a temperature climate. Our estimate of glomerular filtration, however, is similar to published data for Caucasians. The difference in proximal tubular function may reflect possible renal adaptation to a hot, humid climate. We conclude that renal function of tropical residents differs from that of residents of a temperate climate. This difference may be due to renal adaptation to the hot, tropical climate. Received: 1 July 1996 / Revised: 22 December 1996 / Accepted: 8 January 1997     

Daily rhythms of swimming activity,synchronization to different feeding times and effects on anesthesia practice in an Amazon fish species (Colossoma macropomum)

ABSTRACT

This study aimed to investigate the existence of day-night differences in the time for anesthesia and recovery in tambaqui exposed to the anesthetic eugenol and the influence of feeding time. Thus, we evaluated: (1) swimming activity; (2) food anticipatory activity (FAA) as a synchronizer of swimming activity and change to susceptibility to anesthetic; and (3) the effects of diurnal/nocturnal anesthesia exposure of fish feeding in the mid-light phase: 12:00 h (ML) and fish feeding in the mid-dark phase: 00:00 h (MD). Our findings revealed strictly nocturnal activity for tambaqui (94.2%), known as diurnal fish to date. Moreover, FAA was observed in tambaqui fed at MD, which showed a sustained increase in activity that began 2 h before feeding time and lasted until feeding. In contrast, no FAA was observed in fish fed at ML. Regarding anesthesia by day or night, the tambaqui treated with eugenol exhibited no difference in induction time. However, differences were observed in recovery times, with fish anesthetized at day recovering in 1–2 min and fish anesthetized at night recovering in 5–7 min. In short, our findings revealed for the first time the nocturnal behavior of tambaqui. These results indicated that recovery by day/night by eugenol in tambaqui has a strong dependence of behavioral patterns and the time of day.     

CHRONOTYPE,SLEEP LOSS,AND DIURNAL PATTERN OF SALIVARY CORTISOL IN A SIMULATED DAYLONG DRIVING

The study focused on chronotype-related differences in subjective load assessment, sleepiness, and salivary cortisol pattern in subjects performing daylong simulated driving. Individual differences in work stress appraisal and psychobiological cost of prolonged load seem to be of importance in view of expanding compressed working time schedules. Twenty-one healthy, male volunteers (mean?±?SD: 27.9?±?4.9 yrs) were required to stay in semiconstant routine conditions. They performed four sessions (each lasting ～2.5?h) of simulated driving, i.e., completed chosen tasks from computer driving games. Saliva samples were collected after each driving session, i.e., at 10:00–11:00, 14:00–15:00, 18:00–19:00, and 22:00–23:00?h as well as 10–30?min after waking (between 05:00 and 06:00?h) and at bedtime (after 00:00?h). Two subgroups of subjects were distinguished on the basis of the Chronotype Questionnaire: morning (M)- and evening (E)-oriented types. Subjective data on sleep need, sleeping time preferences, sleeping problems, and the details of the preceding night were investigated by questionnaire. Subjective measures of task load (NASA Task Load Index [NASA-TLX]), activation (Thayer's Activation-Deactivation Adjective Check List [AD ACL]), and sleepiness (Karolinska Sleepiness Scale [KSS]) were applied at times of saliva samples collection. M- and E-oriented types differed significantly as to their ideal sleep length (6 h 54 min?±?44 versus 8 h 13 min?±?50 min), preferred sleep timing (midpoint at 03:19 versus 04:26), and sleep index, i.e., ‘real-to-ideal’ sleep ratio, before the experimental day (0.88 versus 0.67). Sleep deficit proved to be integrated with eveningness. M and E types exhibited similar diurnal profiles of energy, tiredness, tension, and calmness assessed by AD ACL, but E types estimated higher their workload (NASA-TLX) and sleepiness (KSS). M types exhibited a trend of higher mean cortisol levels than E types (F?=?4.192, p?<?.056) and distinct diurnal variation (F?=?2.950, p?<?.019), whereas E types showed a flattened diurnal curve. Cortisol values did not correlate with subjective assessments of workload, arousal, or sleepiness at any time-of-day. Diurnal cortisol pattern parameters (i.e., morning level, mean level, and range of diurnal changes) showed significant positive correlations with sleep length before the experiment (r?=?.48, .54, and .53, respectively) and with sleep index (r?=?.63, .64, and .56, respectively). The conclusions of this study are: (i) E-oriented types showed lower salivary cortisol levels and a flattened diurnal curve in comparison with M types; (ii) sleep loss was associated with lower morning cortisol and mean diurnal level, whereas higher cortisol levels were observed in rested individuals. In the context of stress theory, it may be hypothesized that rested subjects perceived the driving task as a challenge, whereas those with reduced sleep were not challenged, but bored/exhausted with the experimental situation. (Author correspondence: mmoginsk@cyf-kr.edu.pl)     

Bright-light exposure during daytime sleeping affects nocturnal melatonin secretion after simulated night work

The guidelines for night and shift workers recommend that after night work, they should sleep in a dark environment during the daytime. However, staying in a dark environment during the daytime reduces nocturnal melatonin secretion and delays its onset. Daytime bright-light exposure after night work is important for melatonin synthesis the subsequent night and for maintaining the circadian rhythms. However, it is not clear whether daytime sleeping after night work should be in a dim- or a bright-light environment for maintaining melatonin secretion. The aim of this study, therefore, was to evaluate the effect of bright-light exposure during daytime sleeping on nocturnal melatonin secretion after simulated night work. Twelve healthy male subjects, aged 24.8 ± 4.6 (mean ± SD), participated in 3-day sessions under two experimental conditions, bright light or dim light, in a random order. On the first day, the subjects entered the experimental room at 16:00 and saliva samples were collected every hour between 18:00 and 00:00 under dim-light conditions. Between 00:00 and 08:00, they participated in tasks that simulated night work. At 10:00 the next morning, they slept for 6 hours under either a bright-light condition (>3000 lx) or a dim-light condition (<50 lx). In the evening, saliva samples were collected as on the first day. The saliva samples were analyzed for melatonin concentration. Activity and sleep times were recorded by a wrist device worn throughout the experiment. In the statistical analysis, the time courses of melatonin concentration were compared between the two conditions by three-way repeated measurements ANOVA (light condition, day and time of day). The change in dim light melatonin onset (ΔDLMO) between the first and second days, and daytime and nocturnal sleep parameters after the simulated night work were compared between the light conditions using paired t-tests. The ANOVA results indicated a significant interaction (light condition and3 day) (p = .006). Post hoc tests indicated that in the dim-light condition, the melatonin concentration was significantly lower on the second day than on the first day (p = .046); however, in the bright-light condition, there was no significant difference in the melatonin concentration between the days (p = .560). There was a significant difference in ΔDLMO between the conditions (p = .015): DLMO after sleeping was advanced by 11.1 ± 17.4 min under bright-light conditions but delayed for 7.2 ± 13.6 min after sleeping under dim-light conditions. No significant differences were found in any sleep parameter. Our study demonstrated that daytime sleeping under bright-light conditions after night work could not reduce late evening melatonin secretion until midnight or delay the phase of melatonin secretion without decreasing the quality of the daytime sleeping. Thus, these results suggested that, to enhance melatonin secretion and to maintain their conventional sleep–wake cycle, after night work, shift workers should sleep during the daytime under bright-light conditions rather than dim-light conditions.     

Variability in the Period of the Blood Pressure Orcadian Rhythm in Human Beings

In studies and assessments of human beings done in natural settings, it is assumed that the period τ of circadian rhythms, including ones of systolic (SBP) and diastolic (DBP) blood pressure, is equal to 24 hours. To test this hypothesis, SBP and DBP rhythms were studied in 112 medication-free, non-hospitalized subjects (62 males, 47.1 + 2.0 years [χ ± SEM], and 50 females, 54.5 ± 2.1 years) by 48h ambulatory blood pressure monitoring (ABPM). Of these, 26 were hypertensive (diurnal SBP> 140 mmHg and diurnal DBP> 90 mmHg) and 86 normotensive. All subjects were synchronized by their habitual daytime activities from ?08:00h to ?23:00h ± lh and by sleep at night. The BP was assessed at 15-minute intervals during a continuous 48h span using a Spacelabs model #90207 ABPM. The time series data of each subject were individually evaluated by power spectra analysis for the prominent x of the SBP and DBP rhythms. The prominent X differed from 24 hours in 22/112 subjects for SBP and in 16/112 subjects for DBP. Generally, in these individuals the τ was less than 24 hours. The occurrence of non-24h τ's was more frequent in hypertensive than normotensive subjects; the difference between the groups in the distribution of the prominent τ's by class (τ = 24h, >=12, 12h<24h, etc.) was statistically significant (χ² test =19.1; p < 0.001). No difference in the distribution of x's of blood pressure was detected according to the subject's age and gender. These findings suggest that ABPM done only for a duration of 24h may be too short to characterize accurately the features of the day-night variation in human BP, including the precise period of its rhythm. (Chronobiology International, 14(3), 307–317, 1997)     

Stereoselective pharmacokinetics of methadone in beagle dogs

The pharmacokinetics of methadone were studied in beagle dogs (n = 4) following intravenous administration of the racemate (0.5 mg/kg) and of the individual (R)-(0.25 mg/kg) and (S)-enantiomers (0.25 mg/kg) using a stereospecific HPLC assay. There was no significant difference between the pharmacokinetic parameters of (R)-methadone and (S)-methadone following administration of the individual enantiomers. Stereoselective differences were evident following administration of the racemate (P values for differences in AUC and CL were 0.001 and 0.046, respectively) and the clearance of the (S)-enantiomer was increased when administered as part of the racemate (316 ± 81 vs 487 ± 128 ml/min, P = 0.04). The data suggest that stereoselective disposition including potential enantiomer–enantiomer interactions should be considered in pharmacokinetic–pharmacodynamic studies of (R,S)-methadone. © 1994 Wiley-Liss, Inc.