XIIIth INTERNATIONALCOURSE ON CHRONOPHARMACOLOGYRuprecht-Karls-Universität HeidelbergSummer School 2012: 30 July – 4 August

Although considerable evidence implicates involvement of nitric oxide (NO) in circadian regulation, little is known about possible 24h variations in basal NO metabolism. In this study, daily variations in serum nitrite levels were studied in locally bred mice and rats during the months of September and October. The serum was separated from blood samples obtained at six different times of the day and night (lh, 5h, 9h, 13h, 17h, and 21h after lights off [HALO] from male albino mice and rats). As an index of in vivo NO generation, serum nitrite levels (determined by the diazotization method) in rats exhibited significant temporal fluctuation (unpaired Student t test), with the concentration highest at 5 HALO and 21 HALO and lowest at 9 HALO. No such temporal variation was detected in mice in these studies conducted on locally bred animals in the autumn. (Chwnobiology International, 16(4), 527–532, 1999)     

23个实验小鼠和大鼠近交系的保种研究

从1985年开始,陆续从国外实验动物著名机构引进常用近交系小鼠共23个品系(亚系和亚群)。在多年来保种繁殖过程中,根据修饰平行线系统选择留种动物,同时对动物进行全兄妹近亲交配繁殖,保持完整的谱系记录,控制饲养环境条件,成功地保持了绝大多数品系。同时对多数品系的繁殖性能和体重增长进行了观察,为这些品系的实验应用提供基本参数。     

Spatial and Temporal Variation in the Ant Occupants of a Facultative Ant-Plant1

Striking variation in ant occupation of a facultative ant-plant, Conostegia setosa (Melastomataceae), was found at three scales: local spatial, geographic, and temporal. C. setosa provides housing for ants and grows in groups of stems (clones). The ant occupants of 14 C. setosa clones were censused four times over a 14-mo period at the La Selva Biological Station, Costa Rica, and twice over a 9-mo period at the Nusagandi Station, Panama. Twelve facultative ant species occupied C. setosa stems at La Selva, compared to six facultative and one obligate species at Nusagandi. Occupancy (as % of stems ever occupied/clone) was higher at Nusagandi (median = 89%) compared to La Selva (65%). Occupancy varied among clones at La Selva but not at Nusagandi. C. setosa clones differed between sites, with larger clones and more small stems/clone at La Selva. Occupancy was influenced by clone structure; larger clones contained more ant species at both sites and had lower occupancy at La Selva. Occupancy was highest in larger stems and lowest in small stems at both sites. Temporally, percent occupation/clone did not differ among censuses at either site, but overall occupancy was lower in the dry season at La Selva. Turnover in ant occupants was higher at La Selva than at Nusagandi. The variation observed in this study is likely due to a number of factors, including differences between sites in plant population structure and history, differences between and within sites in ant faunas and their nesting requirements, and changes over space and time in microclimatic variables. Such high variation at multiple scales draws attention to the importance of long-term comparative studies of facultative animal-plant interactions.     

Chronobiologic Evaluation of Angiotensin-Converting Enzyme Activity in Serum and Lung Tissue from Normotensive and Spontaneously Hypertensive Rats

Angiotensin-converting enzyme (ACE) activity in serum and lung tissue from both normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) was determined at six different circadian times. In WKY rats serum ACE varied significantly within 24 h, mainly due to reduced enzyme activity at 12:00 h. In SHR the 24-h profile of serum ACE did not exhibit time-dependent differences. Mean serum ACE activity over 24 h was significantly higher in WKY than in SHR. In lung tissue ACE activity did not depend on the circadian time in either strain. Mean enzyme activity in lung tissue was not different between WKY and SHR. We conclude that circadian changes in the activity of serum and tissue ACE are unlikely to play an important role in the regulation of the circadian blood pressure profile in both normotensive and spontaneously hypertensive rats.     

Chronobiologic Evaluation of Angiotensin-Converting Enzyme Activity in Serum and Lung Tissue from Normotensive and Spontaneously Hypertensive Rats

Angiotensin-converting enzyme (ACE) activity in serum and lung tissue from both normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) was determined at six different circadian times. In WKY rats serum ACE varied significantly within 24 h, mainly due to reduced enzyme activity at 12:00 h. In SHR the 24-h profile of serum ACE did not exhibit time-dependent differences. Mean serum ACE activity over 24 h was significantly higher in WKY than in SHR. In lung tissue ACE activity did not depend on the circadian time in either strain. Mean enzyme activity in lung tissue was not different between WKY and SHR. We conclude that circadian changes in the activity of serum and tissue ACE are unlikely to play an important role in the regulation of the circadian blood pressure profile in both normotensive and spontaneously hypertensive rats.     

大鼠、小鼠辨证的思路与方法

辨证论治是中医学的特色和优势,为开展辨证论治的研究与发展,我国业已发展出百余种证侯动物模型,起到了积极的作用。本文在扼要回顾存在的问题和研究进展后,以气虚证为例,介绍了大鼠和小鼠辨证的思路与方法：（1）人类与大鼠、小鼠气虚证候的比较。研究表明,大鼠、小鼠的气虚表现与人类十分近似,是可以模拟人类气虚辨证的方法和思路,给大鼠、小鼠辨证的。（2）大鼠、小鼠气虚的计量化辨证。标准化和计量化辨证涉及4个基本问题,即如何计量化采集四诊指标、如何计量化辨证、如何确定辨证的标准和阈值,以及如何判断证候及其改善的程度。为此,我们借鉴药理行为学检测技术,比较了悬尾不动、抖笼、旷场、抓力等方法,最终确定旷场和抓力作为气虚证的计量化四诊指标,建立气虚程度/指数计算公式：气虚程度/指数=各动物抓力实测值/正常组均数×0.5＋各动物水平移动实测值/正常组均数×0.3＋各动物直立次数/正常组均数×0.2,以及气虚辨证的入选标准和阈值。通过大量、重复的实验检测与应用,证明以上方法和辨证标准是可行的,而且初步证明气虚证在神经-内分泌-免疫网络有其广泛的基因表达与剪接改变的基础。目前已初步实现大鼠、小鼠常见虚证标准化、计量化的四诊检测和辨证,该方法和技术可以动态跟踪疾病动物证候的演变,评价辨证论治疗效,还可以广泛用于病-证结合的动物实验研究,辨证论治治疗方案的实验优化和评价,中药、中药有效成分和方剂药性的判断等,适用于中医基础、药理、诊断、临床等各中医药学科的研发工作。     

Temporal and Spatial Allozyme Variation in the South American Cactophilic Drosophila antonietae (Diptera; Drosophilidae)

Drosophila antonietae is an endemic South American cactophilic species that uses Cereus hildmaniannus rotting cladodes as breeding sites. We assessed temporal and spatial intrapopulational allozyme variation of two natural populations. Our results suggest that environmental variation (rain precipitation) is probably influencing allozyme temporal variation. Moreover, it seems that D. antonietae does not have intrapopulation structure and has N _ev (variance effective size) 83 and N _ec (number of adult flies that colonize each rotting cladode) = 21. The deficiency of heterozygotes found must be due to null alleles, a temporal Wahlund effect, or selection against heterozygotes. Assortative mating and inbreeding are discarded. This is the first report on allozyme variation in D. antonietae. It gives some insight on intrapopulational genetics through space and time for this species. This is important to understand its general genetic variability and will be essential to future works on the natural history and evolution of this species.     

Differential Effects of Undernourishment and Nutritional Rehabilitation on Serum Leptin Levels in Male and Female Rats

Leptin, a peptide hormone, is secreted by adipose tissue and is crucial to the regulation of feeding behaviour. The present study has shown that both male and female rats which have been undernourished since day six of gestation, show significantly decreased serum leptin levels on postnatal day 12; but when undernourishment continues into adulthood, only males continue to show decreased leptin levels. If nutritional rehabilitation is implemented early enough in males, serum leptin levels recover and nearly reach levels found in control adult males. Undernutrition also has a long term effect on body weight in both sexes, but nutritional rehabilitation leads to some degree of body weight recovery varying with sex and the age at which rehabilitation was implemented. Undernutrition seems to affect different developmental processes in males than in females, with males being more vulnerable than females in so far as long-term effects on serum leptin levels.     

稻草覆盖对云南高原魔芋地温时空变化的影响

连续三年研究稻草覆盖对云南高原山地栽培‘花魔芋’地温时空变化的影响。结果表明：地温具有明显的时空变化特征,与‘花魔芋’的年生长周期各发育阶段的最适温度相吻合。稻草覆盖改变土壤热量交换层(地表层)的性质,具有延缓地温上升和下降的双向动态调控作用,调控幅度随气温升高而增大,随气温下降而加强;增强了同一土层温度的稳定性,其调控幅度随着土壤深度增加而降低,不同观察时间表现出13:00>19:00>7:00的规律。     

Temporal Variation in Hepatic Superoxide Dismutase Activity in Mice

Circadian variations in superoxide dismutase (SOD) activity were determined in liver homogenates of Balb-C mice that were synchronized under controlled environmental conditions with 12 h light: 12 h dark. The activity of hepatic SOD exhibited a significant circadian rhythm, with a minimum at 01:00 h and maximum at 10:00-13:00 h. It is concluded that fluctuations in hepatic SOD activity render mice more susceptible to the toxic effects of reactive oxygen radicals at particular times of the day.     

Temporal Hierarchies of Variation and the Maintenance of Diversity

Abstract A general model shows how the long-term growth rate of a population can be partitioned into components representing various mechanisms of maintenance of species diversity. One component summarises the effects of fluctuation-independent mechanisms, which include classical resource partitioning and frequency-dependent herbivory. Two other components represent fluctuation-dependent mechanisms, the storage effect and relative nonlinearity of competition.
The general model shows how a community will track an equilibrium set by fluctuation-independent mechanisms and the environmental state when community dynamics are faster than the rate of environmental change. Fluctuation-dependent mechanisms can be important for diversity maintenance with or without such tracking, but on long timescales their effects are indistinguishable from those of fluctuation-independent mechanisms.
These considerations lead to a hierarchical view of mechanisms of diversity maintenance where the effects of different timescales are partitioned or merged depending on the timescale of observation. These issues are illustrated with model examples involving various combinations of resource partitioning, fluctuations in recruitment rates, variation in the timing of germination, and seasonality. The very long timescales associated with climate change contain many complexities but nevertheless many ideas applicable to shorter timescales may be useful in a modified form.     

Serum esterase activities and hyperlipidaemia in the streptozotocin -diabetic rat

The levels of activity of four serum esterases were measured in control and streptozotocin-diabetic rats for a period of 6 months. Pseudocholinesterase activity was significantly elevated in the diabetic rats at all points tested, reaching 250% of control activity at 6 months. Levels of paraoxonase activity progressively decreased with time in the diabetic rats, being 36% lower than in controls at 6 months. No significant differences in either serum arylesterase or carboxylesterase activity between control and diabetic rats were observed.     

Taurine Levels in Discrete Brain Nuclei of Rats

Concentrations of taurine have been measured in 44 microdissected rat brain nuclei or areas. Taurine is ubiquitously present and distributed unevenly in the rat brain: the ratio of the highest (pyriform cortex) to lowest (midbrain reticular formation) concentrations is 4.7:1. High taurine levels were found in cerebral cortical areas, caudate-putamen, cerebellum, median eminence, and supraoptic nucleus. Acute pain stress reduced taurine levels in the hypothalamus and the lower brainstem nuclei but not in cortical areas. Increased locomotor and behavioral activities following a high dose of amphetamine elevated taurine concentrations significantly in the substantia nigra and locus ceruleus.     

Temporal patterns of reproduction and outcrossing in weedy populations of Echium plantagineum

Single and multilocus estimates of outcrossing rates were made in three populations of Echium plantagineum. Despite spatial separation, variations in population size (though not density) and reproductive output, no statistically significant difference was detected in outcrossing rates between the populations. Similarly, only slight differences in outcrossing rates were detected within populations when estimates were based on seed collected from flowers open at different times in the flowering season. The earliest flowers tended to have a lower estimated rate of outcrossing. In all cases, multilocus outcrossing rates were high, ranging from 0.81 to 1.05.
In all three populations flower production extended over a period of more than 2 months but the majority of seed was produced by flowers that opened during the first third of the (lowering season. This was largely caused by a high rate of flower production during the early part of the season and not changes in the number of seeds set per flower. The average number of seeds produced per flower varied both between individuals within populations, and between the different populations, but neither of these differences were significant.
Controlled pollination of flowers with self or outcross pollen, applied either singly or together, failed to detect any differences in the likelihood that either type of pollen would give rise to fertile seeds.     

Temporal Diversity Patterns and Phenology in Fruit-feeding Butterflies in the Atlantic Forest

The Atlantic Forest deserves special attention due to its high level of species endemism and degree of threat. As in other tropical biomes, there is little information about the ecology of the organisms that occur there. The objectives of this study were to verify how fruit-feeding butterflies are distributed through time, and the relation with meteorological conditions. Species richness and Shannon index were partitioned additively at the monthly level, and β diversity, used as a hierarchical measure of temporal species turnover, was calculated among months, trimesters, and semesters. Circular analysis was used to verify how butterflies are distributed along seasons and its relation with meteorological conditions. We sampled 6488 individuals of 73 species. Temporal diversity of butterflies was more grouped than expected by chance among the months of each trimester. Circular analyses revealed that diversity is concentrated in hot months (September–March), with the subfamily Brassolinae strongly concentrated in February–March. Average temperature was correlated with total abundance of butterflies, abundance of Biblidinae, Brassolinae and Morphinae, and richness of Satyrinae. The present results show that 3 mo of sampling between September and March is enough to produce a nonbiased sample of the local assemblage of butterflies, containing at least 70 percent of the richness and 25 percent of abundance. The influence of temperature on sampling is probably related to butterfly physiology. Moreover, temperature affects resource availability for larvae and adults, which is higher in hot months. The difference in seasonality patterns among subfamilies is probably a consequence of different evolutionary pressures through time.     

Effect of One Morning Meal and a Bolus of Dexamethasone on 24‐Hour Variation of Serum Leptin Levels in Humans

Objective: We have previously shown that morning administration of dexamethasone in combination with food induces a doubling of serum leptin levels starting at 7 hours after dexamethasone administration, with a maximum effect at 10 hours, the latest time point that we have studied. However, dexamethasone given in the absence of food had no effect on serum leptin at 10 hours. The present experiment was undertaken to determine the duration of the effect of dexamethasone on 24‐hour serum leptin under fasted and fed conditions in humans. Research Methods and Procedures: Six healthy non‐obese male volunteers were studied under the following four conditions: 1) dexamethasone (2 mg intravenously, given at 0900 hours) with fasting; 2) dexamethasone with food (1700 kcal, 55% carbohydrate, 15% protein, and 30% fat, given in one meal 2 hours after dexamethasone administration at 1100 hours); 3) saline with food (same meal); 4) saline with fasting. Serum leptin, glucose, insulin, and cortisol were monitored every 30 minutes for 24 hours. Results: 1) Under the fasting condition, dexamethasone increased leptin nocturnal secretion between 2100 and 2400 hours. 2) A single meal (1700 kcal) at 1100 hours increased nocturnal leptin secretion when compared with the fasting condition. The peak increase of leptin was 123% over baseline between 2100 and 2400 hours, 10 to 14 hours after the meal. 3) In the fed + dexamethasone condition, leptin levels increased from baseline starting 8 hours after dexamethasone injection, reached a maximum increase of 260% between 2100 and 2400 hours, then decreased thereafter, remaining elevated compared to baseline for 16 hours. There was a correlation between 24‐hour leptin secretion and insulin secretion after a single morning meal. Discussion: A single bolus of dexamethasone, given before a single large meal, produces a delayed (6‐hour) but long‐lasting increase in serum leptin (over 16 hours). Under fasted conditions, dexamethasone does not increase daytime leptin but does increase leptin during the night.     

Effects of Repeated Low-dose Soman Exposure on Monoamine Levels in Different Brain Structures in Mice

In order to better understand the effects of repeated low-dose exposure to organophosphorus (OPs) on physiological and behavioural functions, we analysed the levels of endogenous monoamines (serotonin and dopamine) in different brain areas after repeated exposure of mice to sublethal dose of soman. Animals were injected once a day for 3 days with 0.12 LD50 of soman (47 μg/kg, i.p.). They did not show either severe signs of cholinergic toxicity or pathological changes in brain tissue. 24 h after the last injection of soman, inhibition of cholinesterase was similar in plasma and brain (32% and 37% of inhibition respectively). Afterwards, recovery of cholinesterase activity was faster in the plasma than in the brain. Dopamine levels were not significantly modified. On the other hand, we observed a significant modification of the serotoninergic system. An increase of the 5-HIAA/5-HT ratio was maintained for 2 and 4 weeks after exposure in the hippocampus and the striatum respectively. This study provides the first evidence of a modification of the 5-HT turnover in the hippocampus and the striatum after repeated low-dose intoxication with a nerve agent. Further experiments are necessary to evaluate the relationship between these modifications and the unexpected neuropsychological disorders usually reported after chronic exposure of organophosphorus.     

鼎湖山不同演替阶段森林土壤水分时空变异研究

土壤水分作为森林生态系统水分蓄库的主体,森林土壤水分储量及其时空动态与变异对揭示区域植被恢复与气候变化背景下的森林生态系统水文过程响应与服务功能变化机制具有重要意义。本研究以南亚热带地区典型森林植被演替序列马尾松人工林（Pinus massoniana coniferous forest,PF）-马尾松针阔叶混交林（mixed Pinus massoniana/broad-leaved forest,MF）-季风常绿阔叶林（monsoon evergreen broad-leaved forest,MEBF）为研究对象,依托中国生态系统研究网络森林样地建设与监测统一规范对鼎湖山森林生态系统定位站站区内分布的上述森林类型土壤水分的长期定位观测（2005-2015年）,通过分析各演替阶段森林土壤不同土层（0-15、15-30、30-45、45-60、60-75和75-90 cm）土壤体积含水量观测数据,探究该区域森林植被恢复过程中的土壤水分变化及其时空变异。结果表明：在雨热同期且干湿季明显的南亚热带地区,鼎湖山森林土壤储水量及其时间动态受降雨量的影响显著,森林土壤层对降雨具有强烈的调蓄和稳定作用,伴随PF→MF→MEBF自然演替进程,调蓄水分能力逐步增强。林型间,由初期阶段PF到顶级群落MEBF,森林土壤水分储量逐渐提高,且演替后期林型相对于早期林型,土壤储水量均呈现为较小的年际与年内变幅。干、湿季而言,干季时林型间的土壤储水量差异大于湿季,干季时MEBF和MF土壤含水量分别是PF的1.33倍和1.11倍。从土壤含水量的干、湿季期间变异来看,不同林型各土层土壤含水量的变异系数大小均表现为干季大于湿季;垂直剖面方向上,突出表现为无论干湿季MEBF各层土壤含水量变异均比其他两种林型较为缓和,充分体现了MEBF优越的土壤水分时空调配能力。整体上,伴随PF→MF→MEBF自然演替进程,土壤水分储量及其稳定性逐步提升。     

注意缺陷多动障碍儿童睡眠障碍与铁缺乏关系的临床研究

目的：探讨注意缺陷多动障碍（ADHD）患儿睡眠障碍与血清铁蛋白水平的关系。方法：符合美国精神疾病诊断与统计手册第4版（DSM-Ⅳ）中ADHD诊断标准的62例6．14岁ADHD患儿,由ADHD患儿的父母填写睡眠障碍量表（SDSC）,检测ADH-D患儿的血清铁蛋白水平。结果：与血清铁蛋白水平高于45／μg／L的ADHD患儿相比较,血清铁蛋白水平低于45／μg／L的ADHD患儿的SDSC“睡眠醒觉转换障碍”项评分和总分明显高于前者（P〈0．05）。2组间其他项评分比较无显著性差异（P〉0．05）。SDSC“睡眠醒觉转换障碍”项评分与血清铁蛋白水平成负相关（P〈0．05）。结论：血清铁蛋白水平低于451μg／LADHD患儿SDSC“睡眠醒觉转换障碍”（睡眠中的异常运动）的危险性显著增加。