Dynamic interactions between coupled oscillators within the hamster circadian pacemaker.

Within the mammalian suprachiasmatic nucleus, multiple oscillators interact to coordinate circadian rhythms in behavior and physiology. We have developed a behavioral assay that disassociates central oscillators and allows rigorous study of their formal properties and interactions. Rodents held under 24h light:dark:light:dark (LDLD) cycles display “split” activity rhythms that reflect the reorganization of the central pacemaker into two oscillator groups cycling ~12h apart. After transfer to constant conditions, the two activity components rejoin through a series of transients lasting 2–7 days. Here we analyze fusion dynamics, characterize the underlying oscillator interactions, and assess two influencing factors: phase of transfer and lighting conditions upon transfer. Syrian hamsters were split under LDLD with dimly lit nights and then transferred to constant dim illumination or complete darkness during one of the two daily scotophases. Fusion was influenced by phase of transfer, suggesting that the oscillators split under LDLD exert an asymmetric influence over one another. Transfer to constant dim and dark conditions produced similar overall patterns of fusion, but nevertheless differed in the rejoined state of the system. The present results are discussed within a model wherein oscillators influence one another in a phase-dependent manner. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The significance of circadian organization for longevity in the golden hamster

INTRODUCTION: Altered patellofemoral biomechanics may result in pain, instability and early involutive processes. Magnetic Resonance Imaging (MRI), with its panoramic capabilities, has proved to be an effective technique in the study of knee extensor complex changes. The diagnostic advantages of dynamic studies of patellofemoral kinetics are reported in the recent scientific literature. We investigated the diagnostic potentials of passive studies of the knee extensor complex with sagittal and axial cine MRI. Then, we developed and optimized an innovative study method overcoming the limitations of the other dynamic techniques for the correct assessment of patellofemoral biomechanics. MATERIAL AND METHODS: We studied the knee with a .2 T permanent magnet dedicated to the limbs and acquired the images in different positions of flexion-extension with T1-weighted SE and T2-weighted GE sequences. We examined 21 healthy volunteers and 37 of 38 patients with anterior knee joint pain of suspected patellofemoral origin. All the images needed for dynamic studies were acquired in about 20 minutes. For the scan planes not to be affected by patellar motion in the different degrees of knee extension, it is necessary to acquire single axial images to be edited in cine motion afterwards. Each acquisition is aligned along sagittal reference planes depiciting always the same patellar aspect. RESULTS: Significant correlations were found between clinical and cine MR findings in 25 patients. In particular we depicted some extensor complex impingement conditions missed at conventional MRI, which clarified the role played by patellar dysplastic changes in cartilage microtraumas. Our technique was accurate, quite easy to perform and repeatable. We performed cost-effective dynamic studies which were useful in the evaluation of patients with anterior knee pain in whom conventional MRI had failed to provide enough information. CONCLUSIONS: Our technique differs from other passive or active dynamic studies reported on in the literature because the patellar volume does not change during acquisitions. This permits to decrease morphological changes and to simplify, on cine MR reconstructions, the specific analysis of patellofemoral dynamics during flexion-extension. Fewer morphological changes also mean a more accurate analysis showing the role of patellar dysplasia in cartilage microtraumas. Our dynamic MR protocol is accurate, easy to perform and to repeat; it allows dynamic studies in the patients with poor static MR findings.     

The timSL mutant affects a restricted portion of the Drosophila melanogaster circadian cycle

The circadian rhythm genes period (per) and timeless (tim) are central to contemporary studies on Drosophila circadian rhythms. Mutations in these genes give rise to arrhythmic or period-altered phenotypes, and per and tim gene expression is under clock control. per and tim proteins (PER and TIM) also undergo circadian changes in level and phosphorylation state. The authors previously described a period-altering tim mutation, timSL, with allele-specific effects in different per backgrounds. This mutation also affected the TIM phosphorylation profile during the mid-late night. The authors show here that the single amino acid alteration in TIM-SL is indeed responsible for the phenotype, as a timSL transgene recapitulates the original mutant phenotype and shortens the period of perL flies by 3 h. The authors also show that this mutation has comparable effects in a light-dark cycle, as timSL also accelerates the activity offset during the mid-late night of perL flies. Importantly, timSL advances predominantly the mid-late night region of the perL phase response curve, consistent with the notion that this portion of the cycle is governed by unique rate-limiting steps. The authors propose that TIM and PER phosphorylation are normally rate determining during the mid-late night region of the circadian cycle.     

Sex differentiation of t-e circadian system in the golden hamster

The effect of estradiol benzoate (EB) on free-running circadian activity rhythms was studied in gonadectomized hamsters maintained in constant dim illumination. EB shortened the period (tau) of the female, but not of the male circadian activity rhythm. Responsiveness of the circadian system to EB was subject to sexual differentiation. The circadian period of wheel running by female hamsters given a single injection of testosterone propionate on the day of birth did not shorten in response to EB in adulthood. This failure to respond to EB also was observed in normal male hamsters, and was different from the response shown by normal females. Preliminary data suggest that tau of the activity rhythm of males castrated on the day of birth is shortened during EB treatment in adulthood. The differential effects of estradiol on tau are related to anatomic differences between the sexes in neural connections of the substrate for circadian rhythms.     

The photoperiodic response in Syrian hamster depends upon a melatonin-driven circadian rhythm of sensitivity to melatonin

The pineal gland, via the daily pattern of melatonin (MEL) secretion, is directly involved in the conduction of photoperiodic information. The duration of MEL secretion is proportional to the duration of the dark period and, whatever the photoperiod is, MEL synthesis occurs 3 or 4 h after the dark onset in Syrian hamsters. In order to determine the relative importance of the duration or the coincidence hypothesis, a daily infusion protocol was used in sexually active pinealectomized hamsters. Long duration of MEL infusion (10 h) completely inhibit testes whereas short duration infusion (5 h) had no effect. When the animals were infused twice within 2 h 30 min separated by 3 h, they presented a complete gonadal atrophy, similar to the one observed with the 10 h infusion. Measurement of plasma MEL during the infusion and separation periods revealed that MEL reached physiological nighttime values during the infusion period and fell to daytime values 1 h after the end of an infusion period. Thus, the results could not be due to a time additive action of the two MEL pulses. An intermediate response was observed when the 2 signals were applied across the light/dark transition. Gonadal regression did not occur when the 2 periods of infusion were separated by 5 h 30 min. The efficiency of this type of infusion was not dependent on the ambiant photoperiod since similar results were obtained in long and short photoperiods. The infusion was also as effective during the day as well as during the night. These results suggest that there is a rhythm of sensitivity to MEL, based on the coincidence hypotheses, that are important for transmission of photoperiodic information. This rhythm of sensitivity to MEL seems to be entrained by MEL itself, since the efficiency of the two pulses of MEL is not dependent of time of application and/or of photoperiod.     

The pineal and circadian rhythms of temperature selection and locomotion in lizards

The existence of a circadian rhythm of behavioral temperature selection has been demonstrated in lizards (Podarcis sicula) held on a thermal gradient in constant darkness. This rhythm becomes temporarily abolished during 1 week following parietalectomy and 2-3 weeks following pinealectomy. Parietalectomy does not affect the locomotor rhythm, while pinealectomy invariably lengthens the freerunning period of this rhythm. These results support the contention of separate control systems for the temperature selection rhythm and the locomotor rhythm. As neither rhythm is definitively abolished by parietalectomy and pinealectomy, other pacemaking components exist elsewhere in the circadian system of Podarcis sicula which can control both rhythms.     

Effects of corticotropin-releasing factor on circadian locomotor rhythm in the golden hamster

Stress produces a reduction in the amplitude of some circadian rhythms. The neurochemical mechanisms underlying stress-induced changes in circadian rhythms are not known. To investigate a possible role of corticotropin-releasing factor (CRF) in this phenomenon, three related experiments were carried out: activity rhythms of male golden hamsters (10/14 hours light/dark entrained, lights on at 0800 h) were measured 1) following the intracerebroventricular administration of CRF (0.5, 1.0, 2.0, or 4.0 microg) at two different times of day, 2) following social stress (30-min resident-intruder confrontation), 3) and following the administration of the CRF-antagonist alpha-helical CRF9-41 (2.0 microg) prior to a 15-min resident-intruder confrontation. CRF produced a significant, dose-related decrease in circadian rhythm amplitude following administration in the morning hours, but not in the afternoon. CRF also induced transient increases in activity post injection concomitant with an activation of the hypothalamic-pituitary-adrenocortical (HPA) system. Stress similarly reduced the amplitude of activity patterns and stimulated the HPA system. The stress-induced depression of circadian rhythm amplitude was significantly attenuated following alpha-helical CRF9-41. These data suggest a role for CRF in the stress-related modulation of circadian locomotor rhythm amplitude.     

The kakapo mutation affects terminal arborization and central dendritic sprouting of Drosophila motorneurons

The lethal mutation l(2)CA4 causes specific defects in local growth of neuronal processes. We uncovered four alleles of l(2)CA4 and mapped it to bands 50A-C on the polytene chromosomes and found it to be allelic to kakapo (. Genetics. 146:275- 285). In embryos carrying our kakapo mutant alleles, motorneurons form correct nerve branches, showing that long distance growth of neuronal processes is unaffected. However, neuromuscular junctions (NMJs) fail to form normal local arbors on their target muscles and are significantly reduced in size. In agreement with this finding, antibodies against kakapo (Gregory and Brown. 1998. J. Cell Biol. 143:1271-1282) detect a specific epitope at all or most Drosophila NMJs. Within the central nervous system of kakapo mutant embryos, neuronal dendrites of the RP3 motorneuron form at correct positions, but are significantly reduced in size. At the subcellular level we demonstrate two phenotypes potentially responsible for the defects in neuronal branching: first, transmembrane proteins, which can play important roles in neuronal growth regulation, are incorrectly localized along neuronal processes. Second, microtubules play an important role in neuronal growth, and kakapo appears to be required for their organization in certain ectodermal cells: On the one hand, kakapo mutant embryos exhibit impaired microtubule organization within epidermal cells leading to detachment of muscles from the cuticle. On the other, a specific type of sensory neuron (scolopidial neurons) shows defects in microtubule organization and detaches from its support cells.     

烧结温度对Fe-Ni-Mo磁温度补偿合金热磁性能的影响

采用粉末冶金法制备Fe-Ni-Mo磁温度补偿合金,研究了烧结温度对试样相结构和热磁性能的影响.对不同烧结温度试样X射线衍射(XRD)分析发现:1100℃与1150℃烧结产物均为γ(Fe, Ni)与体心立方Mo;1200℃为γ(Fe, Ni)、α(Fe, Ni)与体心立方Mo;1250℃烧结产物分别为γ(Fe, Ni)、α(Fe, Ni)、体心立方Mo与面心立方Mo四相;1300℃烧结产物为γ(Fe, Ni)、α(Fe, Ni)与面心立方Mo.从衍射峰相对强度变化可知:烧结产物α(Fe, Ni)含量随烧结温度先增后减,1250℃烧结α(Fe, Ni)含量最高.通过磁性测量与电镜分析可知,粉末冶金法制备磁温度补偿合金的关键是烧结温度.通过粉末冶金法与铸造法制得的试样热磁性能对比可知:粉末冶金法试样的热磁性略低于铸造试样的,但其热磁性能的稳定性得到很大提高.     

The late elongated hypocotyl mutation of Arabidopsis disrupts circadian rhythms and the photoperiodic control of flowering

The dominant late elongated hypocotyl (lhy) mutation of Arabidopsis disrupted circadian clock regulation of gene expression and leaf movements and caused flowering to occur independently of photoperiod. LHY was shown to encode a MYB DNA-binding protein. In wild-type plants, the LHY mRNA showed a circadian pattern of expression with a peak around dawn but in the mutant was expressed constantly at high levels. Increased LHY expression from a transgene caused the endogenous gene to be expressed at a constant level, suggesting that LHY was part of a feedback circuit that regulated its own expression. Thus, constant expression of LHY disrupts several distinct circadian rhythms in Arabidopsis, and LHY may be closely associated with the central oscillator of the circadian clock.     

Maternal entrainment of the developing circadian system in the Siberian hamster (Phodopus sungorus)

The aim of these studies was to investigate maternal entrainment of developing circadian locomotor activity rhythms in the Siberian hamster. In Experiment 1, mothers were transferred from a 16:8 LD cycle into constant dim red light (DD) from the day of parturition, and wheel-running activity of the mother and pups was individually monitored from the time of weaning. The phases of the individual pups' rhythms were found to be synchronized both to the phase of the mother and to the phase of lights off (ZT 12) of the photo cycle that the mother was exposed to until the day of parturition. To investigate whether this synchrony might reflect direct effects of light acting upon the fetal circadian system in late gestation, the experiment was repeated but with mothers placed into DD early in pregnancy (< or = day 7 of gestation). The results were similar to the first study, suggesting that the mother rather than the photo cycle during the latter part of gestation entrains the developing circadian system. The third experiment investigated whether this entrainment occurred during the postnatal period. Breeding pairs were maintained on alternative light-dark cycles, LD and DL, that were 12 h out of phase. Litters born to mothers on one light-dark cycle were exchanged on the day of birth with foster mothers from the reversed light-dark cycle, then raised in DD. Control litters exchanged between mothers from the same light-dark cycle had similar litter synchrony as shown by nonfostered litters of Experiment 1. However, pups cross-fostered with mothers on reversed LD cycles showed a very different distribution of pup phases. Pups were not synchronized to their natural mother but to their foster mother. Moreover, pups were more scattered over the 24-h period and were found to be significantly synchronized to the phase of the reversed LD cycle. These results demonstrate the occurrence of postnatal entrainment in the Siberian hamster. The increased scatter produced by the cross-fostering paradigm results from some litters being completely entrained to the phase of the foster mother, some with an intermediate distribution between the phase of the natural and foster mothers, and a minority being associated with the phase of the natural mother. These results suggest that Siberian hamster pups are initially synchronized either prenatally or at birth but that the mother continues to provide entrainment signals during the postnatal period.     

An NK1 receptor antagonist affects the circadian regulation of locomotor activity in golden hamsters

The objective of this study was to evaluate the impact of a feeding supplementation program on the growth of undernourished children younger than 5 years in the city of Guariba, state of S?o Paulo, Brazil. The sample consisted of 469 malnourished children enrolled in a feeding supplementation program sponsored by the State Health Secretariat. The children were divided into four groups according to how long they had been enrolled in the program: in group 1, the children had been enrolled for up to 12 months; in group 2, from 12 to 24 months; in group 3 from 24 to 36 months; and in group 4 the children had been enrolled for more than 35 months. Percentiles for weight/age, height/age and weight/height were calculated for each child. To assess the impact of the program, reference curves for the anthropometric profile were constructed based on expected variations in population percentiles. The changes observed in each group were analyzed statistically (McNemar). Groups 1 and 2 presented weight recovery and gains in the weight/height ratios for the most severely malnourished children; in group 3, the weight/height ratio was maintained and there was a discrete tendency towards weight recovery, which was reversed in group 4, in which the weight was again low in relation to height. The feeding supplementation program temporarily minimized severe nutritional deficiencies but was not sufficient to recover and maintain normal growth.     

Rhythms of temperature selection and body temperature are out of phase in the golden hamster.

Body temperature, locomotor activity, and thermoregulatory behavior of freely moving golden hamsters maintained in a spatial thermocline were measured over several weeks. The thermoregulatory behavior of temperature selection exhibited a robust daily rhythm 180° out of phase with the rhythms of body temperature and locomotor activity. However, the parameters of the body temperature rhythm (rhythmicity, mean level, and amplitude) were not significantly affected by the thermoregulatory behavior. Although the observed phase difference between the rhythms of temperature selection and body temperature might suggest that thermoregulatory behavior is modulated to oppose (rather than to defend) the rhythm of body temperature, the absence of effect of temperature selection on the parameters of the body temperature rhythm fails to reveal the physiological significance of such opposition. Further studies are necessary to establish the physiological significance of the rhythm of temperature selection. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Homeostatic versus circadian effects of melatonin on core body temperature in humans

Liquid-assisted ventilation, as an alternative ventilation strategy for respiratory distress, is progressing from theory and basic science research to clinical application. Biochemically inert perfluorochemical liquids have low surface tension and high solubility for respiratory gases. From early immersion experiments, two primary techniques for liquid-assisted ventilation have emerged: total liquid ventilation and partial liquid ventilation. While computer-controlled, time-cycled, pressure/volume-limited total liquid ventilators can take maximum advantage of these liquids by completely eliminating the gas phase in the distressed lung, partial liquid ventilation takes advantage of having these liquids in the lung while maintaining gas ventilation. The benefits of both partial and total techniques have been demonstrated in animal models of neonatal and adult respiratory distress syndrome, aspiration syndromes and congenital diaphragmatic hernia and also in combination with other therapeutic modalities including extracorporeal membrane oxygenation, high-frequency ventilation and nitric oxide. Additionally, nonrespiratory applications have expanding potential including pulmonary drug delivery and radiographic imaging. Since its use in neonates in 1989, liquid-assisted ventilation in humans has progressed to a variety of clinical experiences with different aetiologies of respiratory distress. The future holds the opportunity to clarify and optimize the potential of multiple clinical applications for liquid-assisted ventilation.     

Nonphotically induced phase shifts of circadian rhythms in the golden hamster: activity-response curves at different ambient temperatures

Running in a novel wheel during the subjective day can shift the circadian activity rhythm of a hamster. The amount of running is thought to be an important variable. We generated a dose-response (activity-phase shift) curve for the amount of wheel running during a 3 h period starting 8 h before normal dark onset in a 14:10 LD cycle. At room temperature (23 degrees C) the relationship was sigmoidal: from 0 to 4000 revolutions resulted in minimal phase advances (up to 50 min). From 4000 to 5000 revolutions the magnitude of the advances increased sharply, and above 5000 revolutions phase advances were asymptotic at about 3 h. The same general relationship held when hamsters were stimulated to be more active in the novel wheel by lowering the ambient temperature to either 11 degrees C or 6 degrees C. However, at these lower temperatures, a significant number of animals did not shift more than the minimal amount of 50 min even though they ran more than 5000 revolutions. This indicates that running per se in a novel wheel was not sufficient to induce phase shifts. Possibly, at room temperature, the amount of wheel running reflects a particular motivational state produced by the rewarding nature of wheel running, although at low ambient temperatures at least some individuals run primarily to meet thermoregulatory needs.     

The association of nonsense mutation with exon-skipping in hprt mRNA of Chinese hamster ovary cells results from an artifact of RT-PCR

RT-PCR of RNA from CHO cells with nonsense mutations in the hprt gene frequently detects minor hprt mRNA species lacking one or more exons. Many nonsense mutants also contain greatly reduced concentrations of the major, normally spliced hprt mRNA. In this study, we examined the hypothesis that exon-deleted mRNAs are normal constituents of CHO cells, but are not detected in wild-type parental cells and most missense mutants because their amplification is suppressed by relatively high concentrations of normally spliced hprt mRNA. A protocol designed to specifically detect exon-deleted mRNAs was conducted using RNA from parental cells and identified all the exon-deleted species typical of nonsense mutants. Quantitative analysis of parental cell RNA measured these exon-deleted mRNAs at < or = 0.7% of the abundance of the full-sized species. Nonsense and missense mutants had comparable amounts of exon-deleted mRNAs, which varied both above and below parental concentrations. The relative concentrations of particular exon-deleted species could be explained by the location of nonsense mutations remaining in the mRNA or by structural effects of mutations on splicing. Exon-deleted mRNAs were detected by RT-PCR when the concentration of the most abundant exon-deleted species was > or = 2% of the full-length mRNA. This occurred for mutants with nonsense mutations in internal exons. RT-PCR conditions were shown to suppress the amplification of exon-deleted species 40-fold when full-length mRNA was abundant, which occurred for parental lines and missense mutants. Our results verify that RT-PCR conditions can produce an artifactual association between nonsense mutation and exon-skipping when minor, exon-deleted mRNA is relatively enriched.