Five-year changes in ovarian function restoration in premenopausal patients with breast cancer taking tamoxifen after chemotherapy: An ASTRRA study report.

BACKGROUND: Adding ovarian function suppression (OFS) after chemotherapy improves survival in young women with moderate- and high-risk breast cancer. Assessment of ovarian function restoration after chemotherapy becomes critical for subsequent endocrine treatment and addressing fertility issues. PATIENTS AND METHODS: In the adding OFS after chemotherapy trial, patients who resumed ovarian function up to 2 years after chemotherapy were randomised to receive either 5 years of tamoxifen or adding 2 years of OFS with tamoxifen. Ovarian function was evaluated from enrolment to randomisation, and patients who did not randomise because of amenorrhoea for 2 years received tamoxifen and were followed up for 5 years. Prospectively collected consecutive hormone levels (proportion of patients with premenopausal follicle-stimulating hormone [FSH] levels <30 mIU/mL and oestradiol [E2] levels ≥40 pg/mL) and history of menstruation were available for 1067 patients with breast cancer. RESULTS: Over 5 years of tamoxifen treatment, 69% of patients resumed menstruation and 98% and 74% of patients satisfied predefined ovarian function restoration as per serum FSH and E2 levels, respectively. Menstruation was restored in 91% of patients younger than 35 years at baseline, but in only 33% of 45-year-old patients over 5 years. Among these patients, 41% experienced menstruation restoration within 2 years after chemotherapy and 28% slowly restored menstruation after 2-5 years. Younger age (<35 years) at baseline, anthracycline without taxanes and ≤90 days of chemotherapy were predictors of menstruation restoration. CONCLUSIONS: During 5 years of tamoxifen treatment after chemotherapy, two-thirds of the patients experienced menstruation restoration, especially patients younger than 35 years. Young age, Adriamycin without taxanes and short duration of chemotherapy appeared to have a positive effect on ovarian reserves in the long term. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT00912548.

Acotiamide improves stress-induced impaired gastric accommodation.

BACKGROUND: Gastric accommodation is a reflex reaction related to gastric reservoir function. Psychological stress, such as anxiety, inhibits gastric accommodation in humans. Acotiamide enhances the effect of acetylcholine in the enteric nervous system, enhances gastric contractility, and accelerates delayed gastric emptying. However, the effect of acotiamide on stress-induced impaired gastric accommodation remains unclear. Therefore, we examined the effect of acotiamide on gastric accommodation and stress-induced impaired gastric accommodation using a conscious guinea pig model. METHODS: A polyethylene bag was inserted through the distal region of the gastric body into the proximal stomach of 5-week-old male Hartley guinea pigs. Gastric accommodation was evaluated by measuring the intrabag pressure in the proximal stomach after oral administration of a liquid meal. In the stress model, animals were subjected to water-avoidance stress. Acotiamide (Z-338) or nizatidine was administered subcutaneously. Fecal output was determined as the number of fecal pellets. KEY RESULTS: Administration of the liquid meal significantly decreased intrabag pressure, indicating induction of gastric accommodation. Acotiamide treatment prolonged liquid meal-induced gastric accommodation and significantly increased the number of fecal pellets compared to controls. Water-avoidance stress significantly inhibited liquid meal-induced gastric accommodation. Pretreatment with acotiamide significantly improved stress-induced impaired gastric accommodation. The number of fecal pellets in the acotiamide group increased significantly compared to controls. Acotiamide, but not nizatidine, significantly decreased gastric emptying. CONCLUSIONS & INFERENCES: Acotiamide prolongs gastric accommodation and improves stress-induced impaired gastric accommodation, indicating a potential role for acotiamide in the treatment of functional dyspepsia through its effects on gastric accommodation reactions.

Tyramine-immunoreactive neuronal structures in the rat brain: abundance in the median eminence of the mediobasal hypothalamus.

Immunoreactivity to p-tyramine, one of the natural trace amines, was studied in the rat brain by an anti-p-tyramine antibody. Immunoreactivity to this amine is very weak in the nigrostriatal dopaminergic neurons and terminals, and weak in the locus coeruleus noradrenergic ones. It was intensified in these structures after monoamine oxidase inhibition. On the other hand, this amine was highly concentrated in the median eminence of the mediobasal hypothalamus, in which its physiological function on prolactin release has been demonstrated.

Influence of sleep disturbance on steroid 5alpha-reductase mRNA levels in rat brain.

Sleep deprivation has been shown to affect the production of steroid hormones in peripheral steroidogenic organs, but little is known about the influence of sleep disturbance on the metabolism of steroid hormones in the brain. To elucidate a possible association of the sleep-wake cycle with brain neurosteroid metabolism, the influence of short-term sleep disturbance on the expression of mRNA encoding steroid 5alpha-reductase, the enzyme converting progesterone and other steroid hormones to their neuroactive 5alpha-reduced metabolites, was investigated. Rats were first subjected to non-selective disturbance of the sleep-wake cycle, and the expression of steroid 5alpha-reductase mRNA in rat hippocampus and brainstem was determined using a semi-quantitative one-step RT-PCR technique. Non-selective disturbance of the sleep-wake cycle resulted in the elevation of 5alpha-reductase mRNA levels in the brainstem, but not in the hippocampus, and the elevated mRNA expression returned to the basal levels after a short period of the sleep recovery. Further studies showed that selective REM sleep deprivation significantly elevated 5alpha-reductase mRNA levels in both hippocampus and brainstem, thus proposing the possibility that REM sleep reduction may largely contribute to the elevation of steroid 5alpha-reductase mRNA levels observed during short-term disturbance of the sleep-wake cycle. Since the enhancement of steroid 5alpha-reductase gene expression may result in the elevation of neuroactive 5alpha-reduced steroid production in the brainstem, the findings presented here provide further evidence for suggesting that neuroactive steroids may play a physiologically important role in the neuronal network for REM sleep initiation and maintenance.

Increase of hippocampal acetylcholine release at the onset of dark phase is suppressed in a mutant mice model of evening-type individuals.

We have previously reported that clock mutant mice on Jcl:ICR background show about 2-h delayed circadian profiles in body temperature, spontaneous activity and sleep-wake rhythm, and that they appear to be useful as a model of evening-type of individual. Hippocampal acetylcholine (ACh) release which is positively correlated with attention, learning and memory shows a circadian variation. In this study, changes in hippocampal ACh release in transitional phase from light (rest) to dark (active) period in clock mutant mice were monitored using an in vivo microdialysis method. Compared with wild mice, the increase in hippocampal ACh in the first 2 h of the active period in the mutant mice was suppressed in parallel with peak frequency in electroencephalogram theta rhythm. The molecular basis of the circadian system appears to have a strong effect on hippocampal cholinergic function, and is probably associating with individual temporal differences in voluntary behavior, cognition, learning and/or memory performance.

Acceleration of EEG theta wave precedes the phasic surge of arterial pressure during REM sleep in the rat.

We investigated temporal relationships among EEG theta frequency, eye movement (EM) burst and phasic surge of arterial pressure (AP) during rapid eye movement (REM) sleep in six rats. Changes in EEG theta frequency, EM number, mean AP (MAP) and heart rate (HR) during 1 s bins were obtained with reference to the onset of the EM bursts. The acceleration of EEG theta frequency preceded both the EM burst and the phasic increases of AP. The increase of MAP was closely correlated with the theta frequency but not the EM number during the period of EM burst. The possible involvement of the limbic system for the generation of phasic surge in AP during REM sleep is discussed.

Mouse model for morningness/eveningness.

Human morning/evening preferences has recently been reported to be associated with polymorphism of the 3' flanking region of the Clock gene, which was the first identified mammalian circadian clock gene. We recorded body temperature, spontaneous activity, electroencephalogram and electromyogram for 48 h in mice with Jcl:ICR genetic background and homozygous for the Clock mutation (Cl/Cl on Jcl:ICR). In both wild-type and Cl/Cl on Jcl:ICR, body temperature, activity, wake and sleep were completely entrained to LD cycle. However, phases of the rhythm for body temperature, activity and wake duration in the Cl/Cl on Jcl:ICR were about 2 h delayed in comparison with the wild-type. This study has provided further evidence on the close relationship between human morning/evening preference and the molecular basis of circadian clock system, and has suggested that Cl/Cl on Jcl:ICR is useful for an animal model for human morning/evening preference.

Spectral analyses of PGO-on neurons during paradoxical sleep in freely moving cats.

We have investigated the power spectral density (PSD) characteristics of spontaneous neuronal activities during paradoxical sleep (PS), including phasically firing PGO-on and tonically discharging type I-R neurons in the pontine tegmentum. The PSD of lateral geniculate PGO waves and blood pressure (BP) were also examined. During PS, the PSD of PGO-on neurons and PGO waves showed an almost white noise-like pattern, while that of TI-R neurons and BP exhibited a 1/f spectrum. It appears, therefore, that the neuronal and autonomic slow fluctuations found in the 1/f spectrum during PS are not driven by the PGO wave system.

Differential effect of short-term REM sleep deprivation on NGF and BDNF protein levels in the rat brain.

It is well known that REM sleep is associated with memory consolidation, especially, procedural skill learning. Neurotrophic factors are known to be involved in synaptic plasticity. We therefore investigated the effects of selective REM sleep deprivation (RSD) on NGF and BDNF proteins in the hippocampus, cerebellum and brainstem in the rat. NGF and BDNF were detected by an ELISA. Our findings show that 6 h RSD affected the NGF and BDNF protein levels in different manner. In the cerebellum and brainstem, BDNF was significantly decreased, while NGF was not changed. Conversely, in the hippocampus, NGF was significantly decreased while BDNF was not changed. This study indicates that REM sleep may be associated with the secretion of neurotrophic factors and thus contribute to the memory functions.

Diurnal changes in pineal extracellular indoles of freely moving rats.

We observed 24 h changes of 'extracellular' indoleamines in pineal microdialysates taken from freely moving rats under a 12 h light/12 h dark cycle. Pineal indoleamines in microdialysates were analyzed by high-performance liquid chromatography with electrochemical detection. Consequently, they exhibited marked diurnal variations. The diurnal level in the extracellular 5-HT, higher for the light and lower for the dark, was parallel to that in 5-hydroxyindoleacetic acid and diametrically opposite to that in N-acetylserotonin (NAS). The release of 5-HT into the extracellular space transiently increased for the first 2 h after the dark onset and then became lower during the dark.

Effects of an eight-hour advance of the light-dark cycle on sleep-wake rhythm in the rat.

We observed effects of an 8-h advance of the light-dark (LD) cycle on the sleep-wake rhythm in the rat. On the day phase-advanced, rapid eye movement (REM) sleep increased with its enhanced diurnal amplitude. Non-REM (NREM) sleep gradually increased in parallel with the decrease of its diurnal amplitude. Although the acrophase of NREM sleep gradually advanced after the phase advance, that of REM sleep did not significantly change. We confirmed that diurnal rhythm of REM sleep was hardly shifted and dissociated from the rhythm of NREM sleep under the eight-hour advance of the LD cycle in the rat.

Pineal microdialysis in freely moving rats.

We describe a surgical technique to implant the guide cannula for in vivo microdialysis in the rat pineal gland. This technique has the following features and advantages: (a) does not require ligation of the superior or transverse sinus, (b) minimizes bleeding from the dural veins, (c) does not disturb the sympathetic innervation originating from superior cervical ganglia, which is essential for pineal function. This new technique makes it possible to carry out chronic pineal microdialysis of freely moving rats.

Effect of decerebration on blood pressure during paradoxical sleep in cats.

We investigated the effects of decerebration on long-term variations in arterial blood pressure during paradoxical sleep (PS) in cats. In normal cats, the blood pressure decreased during the transition from slow wave sleep to PS and maintained its lower level throughout PS for several days after surgery. After this early postoperative stage, however, the arterial hypotension was replaced by tonic and phasic rises in blood pressure during PS. Such long-term changes in blood pressure were completely abolished when the brain stem was transected at the ponto-mesencephalic junction, and the cats consistently exhibited a sustained fall in blood pressure throughout the survival periods of 1 month or more.

Semi-quantitative RT-PCR analysis of environmental influence on P450scc and PNMT mRNA expression in rat adrenal glands.

Environmental influence on brain function, particularly spatial learning and memory, has been extensively investigated, but little is known about the influence of environmental conditions on the functions of peripheral organs. In the present study, the effects of different housing conditions on the steady-state levels of mRNAs encoding cholesterol side-chain cleavage enzyme (cytochrome P450scc) and phenylethanolamine N-methyltransferase (PNMT) in adrenal glands was examined to investigate the environmental influence on both adrenocortical and adrenomedullary functions. Behavioral changes of the animals housed in different conditions were first examined to assess the relevance of environmental manipulation used. In consistent with previous findings, housing of the animals in enriched conditions resulted in the significant reduction of spontaneous motor activity (locomotor activity and rearing) in comparison with housing in isolated conditions, thus indicating the relevance of housing conditions used in this work for investigating the environmental influence on adrenal function. Then, the effects of these housing conditions on P450scc and PNMT mRNA levels in adrenal glands were examined using semi-quantitative RT-PCR method. In comparison with the isolated group, the enriched group showed significantly higher levels of P450scc mRNA. In contrast, PNMT mRNA levels in the enriched group were significantly lower than those in the isolated group. These results propose the possibility that the environmental conditions may cause differential alterations in adrenocortical and adrenomedullary functions, although their possible association with behavioral changes still remains to be elucidated.

Ultradian rhythm of 100 min in the dark phase EEG of the rat.

Using the "variance in statistics" as an index of electroencephalogram (EEG) parameters, we observed the cortico-hippocampal EEG rhythm under a 12:12-h light-dark condition in the rat with chronically implanted electrodes for EEG recording. The above EEG variance was simply measured in real time and on line through a personal computer. It corresponded to EEG slow wave activity and expressed the process of slow wave sleep as described in the two-process model by Borbély et al. Only in the dark phase, mean power spectral density of the EEG variance had a significant peak at about 1/100 cycles/min. This 100-min rhythmicity similar to the basic rest-activity cycle in human beings was observed in rats, particularly in the dark (active) phase for nocturnal animals. We propose that this ultradian 100-min rhythm is essential for the rat to maintain the waking state dominantly over the 12-h dark period.

Relationship between eye movements and oneiric behavior in cats.

The relation between oneiric behavior and rapid eye movements (REMs) in paradoxical sleep (PS) without muscle atonia was analyzed in cats. Most isolated REMs were related to orienting behavior, whereas most REM bursts were related to generalized body movements (jumping, attacking, etc.). Only isolated, high amplitude REMs had any possibility of corresponding to visual images in dreams. From these findings we propose the existence of both dream-related and nondream-related REMs even in animals.

Spontaneous activity, sleep, and body temperature in rats lacking the CCK-A receptor.

Because of a genetic mutation, the Otsuka-Long-Evans-Tokushima Fatty (OLETF) rat, a model for human non-insulin-dependent diabetes mellitus (NIDDM), shows no expression of the CCK-A receptor gene. We investigated the spontaneous physical activity, sleep, and body temperature in young OLETF rats that had not yet developed diabetes mellitus, and compared these data with age-matched control LETO (non-diabetic strain, Long-Evans-Tokushima-Otsuka) rats. The amount of large movements during the dark phase for the OLETF rats was significantly less than that of control rats. Thus, the amounts of total daily large movement and the ratio of dark-to-light phase movement in the OLETF rats were less than those of control rats, although the amount of small movement was similar for both groups. The diurnal rhythm of body temperature was similar for both groups. In addition, the amount of and circadian rhythm for each vigilance state and slow-wave activity were similar for the two groups. This study demonstrates that the CCK-A receptor might play a role in affecting the level of motor activity, adding hyperphagia, and the circadian rhythm of large movement in these rats prior to the manifestation of NIDDM. In contrast, a CCK-A receptor deficiency does not appear to affect sleep or body temperature in these rats.

Effects of intravenously administered vitamin B12 on sleep in the rat.

Vitamin B12 (VB12) has been reported to normalize the entrainment of circadian rhythms in the non-24-h sleep wake cycle and delayed sleep phase insomnia in humans. The purpose of this work was to clarify whether the peripheral administration of VB12 has any sleep-promoting effect on the sleep-wake rhythm in freely moving rats. After a baseline day of saline infusion. VB12 (500 micrograms/kg/day) was administered continuously for 4 days via the jugular vein. Polysomnographic recordings were carried out concurrently. In both the light and the 24-h periods, the amount of non-rapid eye movement (NREM) sleep increased significantly on VB12-days 2 and 3, while the amount of REM sleep increased significantly on VB12-day 2. In the light period, the increase in NREM sleep was due to increased duration of the episode, while the tendency to an increase in REM sleep was due to an increased number of episodes. Changes in the diurnal sleep-wake rhythm tended to appear in the earlier light period. The serum VB12 concentrations in the VB12 group were 40 times higher than in controls. These findings suggest that peripherally infused VB12 has promoting effects on the rat's sleep, especially in the light period.

Long-term profiles of sleep-related hemodynamic changes in the postoperative chronic cat.

The long-term postoperative changes in mean arterial pressure (MAP) and heart rate (HR) during sleep in cats (n = 4) were observed. In the first or second week after surgery, MAP and HR in slow wave sleep (SS) just prior to paradoxical sleep (PS) were higher than those in PS as described in previous reports. In the fourth or fifth week after surgery, the above relationships of MAP and HR between SS and PS were inverted as demonstrated in the present study. The results show that getting true physiological control data is not so simple but a complex task.

Postoperative changes of rat EEG variance.

Concerning an EEG parameter variance, the long-term postoperative changes were observed in rats chronically implanted with bipolar depth electrodes for EEG recordings. The cortico-hippocampal EEG signals were continuously digitized and their variances were computed for each 1 min in real time. Daily mean of the EEG variance gradually increased over 2 weeks after surgery for the implantation of chronic electrodes and, then, reached a plateau. To observe the absolute quantity of the depth EEG in the rat, at least 2 weeks are required before data collections.