Development of a new tracer-encapsulated solid pellet injection system for more precise control of tracer-impurity-deposit location in LHD.

A new tracer-encapsulated solid pellet (TESPEL) injection system has been developed additionally for the LHD heliotron. This system has three-dimensionally bended guide tubes, which allows us to inject the TESPEL obliquely on a poloidal cross-section of the LHD plasma. Consequently, this system enables us to control a tracer-impurity-deposited location more precisely. Moreover, this system can make it possible for the tracer impurity to be deposited even only inside the ergodic region (outside a last closed flux surface of the LHD plasma). A stereoscopic fast camera system has been also developed for capturing a three-dimensional TESPEL trajectory.

Interspecies differences in the empty body chemical composition of domestic animals.

Domestication of animals has resulted in phenotypic changes by means of natural and human-directed selection. Body composition is important for farm animals because it reflects the status of energy reserves. Thus, there is the possibility that farm animals as providers of food have been more affected by human-directed selection for body composition than laboratory animals. In this study, an analysis was conducted to determine what similarities and differences in body composition occur between farm and laboratory animals using literature data obtained from seven comparative slaughter studies (n = 136 observations). Farm animals from four species (cattle, goats, pigs and sheep) were all castrated males, whereas laboratory animals from three species (dogs, mice and rats) comprised males and/or females. All animals were fed ad libitum. The allometric equation, Y = aX b , was used to determine the influence of species on the accretion rates of chemical components (Y, kg) relative to the growth of the empty body, fat-free empty body or protein weights (X, kg). There were differences between farm and laboratory animals in terms of the allometric growth coefficients for chemical components relative to the empty BW and fat-free empty BW (P < 0.01); farm animals had more rapid accretion rates of fat (P < 0.01) but laboratory animals had more rapid accretion rates of protein, water and ash (P < 0.01). In contrast, there was no difference in terms of the allometric growth coefficients for protein and water within farm animals (P > 0.2). The allometric growth coefficients for ash weight relative to protein weight for six species except sheep were not different from a value of 1 (P > 0.1), whereas that of sheep was smaller than 1 (P < 0.01). When compared at the same fat content of the empty body, the rate of change in water content (%) per unit change in fat content (%) was not different (P > 0.05) across farm animal species and similar ash-to-protein ratios were obtained except for dogs. The fraction of empty body energy gain retained as fat increased in a curvilinear manner, and there was little variation among farm animals at the same fat content of the empty body. These findings may provide the opportunity to develop a general model to predict empty body composition across farm animal species. In contrast, there were considerable differences of chemical body composition between farm and laboratory animals.

Improving TWSTFT short-term stability by network time transfer.

Two-way satellite time and frequency transfer (TWSTFT) is one of the major techniques to compare the atomic time scales between timing laboratories. As more and more TWSTFT measurements have been performed, the large number of point-to-point 2-way time transfer links has grown to be a complex network. For future improvement of the TWSTFT performance, it is important to reduce measurement noise of the TWSTFT results. One method is using TWSTFT network time transfer. The Asia-Pacific network is an exceptional case of simultaneous TWSTFT measurements. Some indirect links through relay stations show better shortterm stabilities than the direct link because the measurement noise may be neutralized in a simultaneous measurement network. In this paper, the authors propose a feasible method to improve the short-term stability by combining the direct and indirect links in the network. Through the comparisons of time deviation (TDEV), the results of network time transfer exhibit clear improved short-term stabilities. For the links used to compare 2 hydrogen masers, the average gain of TDEV at averaging times of 1 h is 22%. As TWSTFT short-term stability can be improved by network time transfer, the network may allow a larger number of simultaneously transmitting stations.

Involvement of NTS2 receptors in stress-induced analgesia.

Stress activates multiple neural systems that suppress pain sensation. This adaptive phenomenon referred as stress-induced analgesia (SIA) is mediated by the activation of endogenous pain inhibitory systems. Both opioid and non-opioid forms of SIA have been elicited in rodents according to stressor parameters and duration. There is accumulating evidence that the endogenous neurotensin (NT) system plays an important role in SIA. Especially, NT-deficient mice were shown to exhibit reduced SIA following water avoidance or restraint stress. Since central NT produces naloxone-insensitive analgesic effects by acting on spinal and supraspinal NTS2 receptors, we hypothesized that NT might mediate non-opioid SIA through NTS2 activation. Here, we evaluated the influence of an opioid-independent severe stress produced by a cold-water swim for 3 min at 15 degrees C on rodent offspring's pain perception. Our results demonstrated that mice lacking NTS2 exhibit significantly reduced SIA following cold-water swim stress. Indeed, NTS2 knockout mice submitted to both acute (plantar test) and tonic (formalin test) pain stimuli show a greater sensitivity to pain in comparison to wild-type littermates. Accordingly, pretreatment with the NT receptor antagonist SR142948A results in a hyperalgesic response to stress induced by cold-water swim. Endogenous NT regulates hypothalamic-pituitary-adrenal axis activity in stress condition by increasing corticosterone plasma levels. Accordingly, the plasma levels of corticosterone measured by radioimmunoassay are significantly reduced in non-stressed and stressed NTS2-deficient mice in comparison with wild-type mice. To further investigate the site of action of NT in mediating SIA, we microinjected NTS2 agonists in lumbar spinal cord and quantified post-stress sensitivity to pain in rats using the plantar test. Exogenously administered NTS2 analogs, JMV-431, beta-lactotensin and NT69L markedly enhance the magnitude and duration of stress antinociception in both 25- and 60-day-old rats. In sum, by using genetic and pharmacological approaches, we demonstrated here that NTS2 receptors mediate non-opioid SIA. Our results also revealed that the release of endogenous NT in response to stress requires the presence of NTS2 to stimulate corticotropin-releasing factor (CRF)-induced elevation of plasma corticosterone, and that NTS2 receptors localized at the lumbar spinal cord participate to the disinhibition of descending pain control pathways. Therefore, these data highlight the significance of NTS2 as a novel target for the treatment of pain and stress-related disorders.

Mouse neurotensin receptor 2 gene (Ntsr2): genomic organization, transcriptional regulation and genetic mapping on chromosome 12.

We earlier isolated and characterized the mouse neurotensin receptor 1 (Ntsr1) gene and developed Ntsr1 null mice. In the present study, we isolated the mouse neurotensin receptor 2 gene (Ntsr2) and characterized the structure. The gene fragments available to us have 1.1 kb of 5' upstream promoter region and a 7 kb coding region composed of four exons. Transcription initiation sites, determined by primer extension analysis, are located at 286 and 303 bp upstream from initiation of the ATG codon. The promoter region contains a TATA-like box, a typical CAAT box and putative GATA-2, CREB, Oct-2 and Ikarous 2 binding elements. We also found novel splice donor-acceptor sites for alternative splicing, which could generate a short form of mRNA encoding a truncate-type receptor. In addition, we determined the chromosomal location of the Ntsr2 gene and mapped it at 6 cM from the centromere on chromosome 12.

Generation and characterization of mice lacking gastrin-releasing peptide receptor.

Gastrin-releasing peptide (GRP) is a mammalian bombesin-like peptide which is widely distributed in the central nervous system as well as in the gastrointestinal tract. GRP binds to its high affinity receptor (GRPR) to elicit a wide spectrum of biological effects on behavior, digestion, and metabolism. To define the in vivo function of GRPR, we generated GRPR null mutant mice by gene targeting. The intracerebroventricular administration of GRP caused hypothermia in wild-type mice, but not in mutant mice. The GRPR deficient mice showed significantly increased locomotor activity during the dark period, and social responses scored by sniffing, mounting, and approaching behaviors against an intruder. Aggressive scores such as fighting and biting were not altered in the mutant mice. These phenotypes were observed in mice generated from two independent ES cell clones and backcrossed to a C57BL/6J background. The GRPR deficient mice should be useful for studying the bombesin system in vivo.

Mice lacking bombesin receptor subtype-3 develop metabolic defects and obesity.

Mammalian bombesin-like peptides are widely distributed in the central nervous system as well as in the gastrointestinal tract, where they modulate smooth-muscle contraction, exocrine and endocrine processes, metabolism and behaviour. They bind to G-protein-coupled receptors on the cell surface to elicit their effects. Bombesin-like peptide receptors cloned so far include, gastrin-releasing peptide receptor (GRP-R), neuromedin B receptor (NMB-R), and bombesin receptor subtype-3 (BRS-3). However, despite the molecular characterization of BRS-3, determination of its function has been difficult as a result of its low affinity for bombesin and its lack of an identified natural ligand. We have generated BRS-3-deficient mice in an attempt to determine the in vivo function of the receptor. Mice lacking functional BRS-3 developed a mild obesity, associated with hypertension and impairment of glucose metabolism. They also exhibited reduced metabolic rate, increased feeding efficiency and subsequent hyperphagia. Our data suggest that BRS-3 is required for the regulation of endocrine processes and metabolism responsible for energy balance and adiposity. BRS-3-deficient mice provide a useful new model for the investigation of human obesity and associated diseases.

[The meaning of eat in home patients with end-stage cancer].

When a patient difficult to eat, he will not be only malnutrition, but loses a volition of his life and falls into depression. This time, we present a patient with relapse advanced pharynx cancer, who could not ingestion. Therefore she had said "DEATH". She tried eagerly rehabilitation of ingestion, swallowing and breathing, and she was able to eat again and could be positive posture. Ingestion is very important for human, and the patient found a meaning of her life and she was able to take home care nursing.

Cloning and characterization of the rat neurotensin receptor gene promoter.

The 5'-terminal region of the rat neurotensin receptor (NTR) gene was isolated and characterized. Genomic Southern analysis revealed that the NTR gene occurs as a single copy in the rat haploid genome. The upstream putative promoter region did not contain canonical TATA or CAAT boxes but has a consensus sequence for the transcription factor Sp1. This promoter is embedded in a large G + C-rich domain with characteristics of an CpG island. Transfection experiments using neurotensin receptor-luciferase fusion genes demonstrated that the 5'-flanking sequence functions as a strong promoter in the NG-108-15 cell. Deletion analysis suggested the presence of a core promoter (-470 to -662) that drives the minimal expression of the NTR gene.

Genomic structure of the mouse gene (Lmnb1) encoding nuclear lamin B1.

The mouse gene (Lmnb1) that encodes nuclear lamin B1 was isolated. Structural analyses revealed that the lamin B1 gene spans about 43 kb of the genome and consists of 11 exons and 10 introns. Exon/intron structure of the B1 gene clearly showed the conserved organization shared among the intermediate filament protein family genes. The presumptive promoter region has high GC content and contains a CAAT box and multiple SP1 sites but no classical TATA box, suggesting that the lamin B1 gene has a typical housekeeping gene promoter with a CpG island. These data reveal the gene structure of the only remaining unanalyzed mouse somatic lamin gene. Gene structures of all the mouse somatic lamins are compared.

Daily walking combined with diet therapy is a useful means for obese NIDDM patients not only to reduce body weight but also to improve insulin sensitivity.

OBJECTIVE: To evaluate the effects of walking combined with diet therapy (1,000-1,600 kcal/day) on insulin sensitivity in obese non-insulin-dependent diabetes mellitus (NIDDM) patients. RESEARCH DESIGN AND METHODS: Subjects were divided into two groups: 10 patients were managed by diet alone (group D), and 14 patients were placed in the diet and exercise group (group DE). Group DE was instructed to walk at least 10,000 steps/day on a flat field as monitored by pedometer (19,200 +/- 2,100 steps/day), and group D was told to maintain a normal daily routine (4,500 +/- 290 steps/day). A glucose clamp procedure at an insulin infusion rate of 40 microU.min-2.min-1 was performed before and after the 6- to 8-week training program. Mean serum insulin concentrations ranged from 720 to 790 pmol/l. RESULTS: While body weight (BW) in groups D and DE decreased significantly (P < 0.01) during the study, the amount of BW reduction in group DE was greater than that in group D (7.8 +/- 0.8 vs. 4.2 +/- 0.5 kg, P < 0.01). After training, glucose infusion rate (GIR) and metabolic clearance rate (MCR) in group D did not significantly increase; however, GIR and MCR increased significantly in group DE, from 17.21 +/- 1.11 to 26.09 +/- 1.11 mumol.kg-1.min-1 (P < 0.001) and from 3.0 +/- 0.3 to 5.3 +/- 0.4 ml.kg-1.min-1 (P < 0.001), respectively. The analysis of variance showed significant effects of exercise (time x exercise, P = 0.0005) for the improvement of MCR. Significant correlations were also observed between delta MCR and average steps per day (r = 0.7257, P < 0.005) in group DE. CONCLUSIONS: Walking, which can be safely performed and easily incorporated into daily life, can be recommended as an adjunct therapy to diet treatment in obese NIDDM patients, not only for BW reduction, but also for improvement of insulin sensitivity.

Regulation of mRNA expression involved in Ras and PKA signal pathways during rat hypoglossal nerve regeneration.

Using in situ hybridization histochemistry and immunohistochemistry, the present study examines the cooperative regulation of transcription of molecules involved in the Ras-signal and the cAMP dependent protein kinase (PKA) pathways during peripheral nerve regeneration in rats. Injury to hypoglossal motor neurons resulted in an increase in extracellular regulated kinase (ERK, or MAP kinase) and ERK kinase (MEK, or MAP kinase kinase) mRNAs, but in a decrease in the expression of the catalytic subunits of PKA (C alpha and C beta) mRNAs. These results show the importance of the Ras-signal pathway in the nerve regeneration process and extend recent observation which suggested a cross-talk between the Ras and PKA pathways in vitro. The down-regulation of PKA may facilitate the activation of the Ras pathway which is located downstream of the growth factor receptor. The present study may suggest a possibility of regulatory talk between these two major signal transduction pathways.

[Effects of ingestion of "BON-NARINE" on immune functions in mice].

A study was carried out to examine the effects of ingestion of "BON-NARINE" (BN) on mice immune functions. Mice aged 12 weeks were divided into 4 groups. The first group was given BN at 30 mg/kg (BN-30 group), the second group was given BN at 90 mg/kg (BN-90 group), the third group was given zymosan at 50 mg/kg (Zy group) and the fourth group was a control receiving no treatment. The mice of groups BN-30 and BN-90 were given BN p.o. at doses of 30 mg/kg and 90 mg/kg per day for 20 consecutive days, respectively. The mice of group Zy were given zymosan i.p. at a dose of 50 mg/kg per day for 2 consecutive days. The results obtained were as follows: 1) Potentiation of phagocytic function of the reticuloendothelial system, examined by the carbon clearance method, was seen in the BN-30, BN-90 and Zy groups. 2) The glucose consumption of peritoneal macrophages (M phi) increased significantly in the BN-30 and Zy groups, but not in the BN-90 group. 3) Superoxide anion (O2-) production of peritoneal M phi significantly increased in the BN-30 and Zy groups compared with the control group, but an increasing tendency was observed in the BN-90 group. 4) The acid phosphatase (APH), beta-glucuronidase (GLU) and lactate dehydrogenase (LDH) activities of peritoneal M phi increased significantly in the BN-30, BN-90 and Zy groups. 5) The proliferation of splenocytes induced by Con A in the BN-30, BN-90 and Zy groups significantly increased compared with the control group. These results demonstrated that the ingestion of "BON-NARINE" promotes phagocytic activity in the reticuloendothelial system in mice and has a stimulatory effect on M phi because of increases in glucose consumption, O2- production, APH, GLU and LDH activities in the peritoneal M phi of mice. BN also intensified the T-cell function represented by Con A-induced splenocyte proliferation.

Growth factors and extracellular signal-regulated kinases (mitogen-activated protein kinase) in the rat pineal gland.

The present study revealed that the pineal gland expressed basic fibroblast growth factor (bFGF) and FGF-receptor 1 (FGFR1/flg), suggesting that bFGF in the pineal gland acts in an autocrine or paracrine manner, which is mediated by FGFR1/flg. The present study also examined gene expression of the extracellular signal-regulated kinase (ERK) family (ERK1-3) which may be intracellular signal mediators of growth factors. ERK1 [mitogen-activated protein kinase (MAP-kinase)] was strongly expressed throughout the pineal gland, while expression of ERK2 and ERK3 was not found. These findings suggest the presence of a signal pathway from bFGF to ERK1 via FGFR1/flg in the pineal gland.

Marginal topography of neurons expressing the substance P receptor in the rat suprachiasmatic nucleus.

Neurons expressing the substance P (SP) receptor (NK1 receptor) in the suprachiasmatic nucleus of the hypothalamus (SCN) have been topographically identified using radioactive in situ hybridization histochemistry. In the anterior hypothalamic area, clustered labeled neurons of small size and exhibiting low levels of gene expression are observed exclusively at the dorsolateral margin of the SCN, straddling cytoarchitectural boundaries of the nucleus. The marginal topography of neurons putative target of a SP-containing retinal input to the ventral SCN indicates that their dendrites bearing the receptor extend towards the retinorecipient part of the nucleus, where they can be modulated by overlapping inputs from the intergeniculate leaflet and the raphe. Eventual interactions between glutamatergic and putative tachykininergic retinal pathways for a coherent photic control of circadian rhythms may therefore occur mainly via intrinsic neuronal connections between their distinct target populations. In addition, since glutamate and SP induce electrophysiological responses in ventrolateral neurons with no interactive effect, neurons integrating both chemical signals, subsequently to their modulation by several influences, may be not located within the ventrolateral SCN. Alternatively but not exclusively, marginal neurons could be a target of SP-containing neurons within the SCN or nearby the nucleus, or from ascending projections from the raphé where serotonin and SP colocalize. The marginal topography of neurons expressing the SP receptor supports the view of the involvement of neurons located in the vicinity of the nucleus in the regulation of circadian rhythms.

Cellular localization of serotonin transporter mRNA in the rat brain.

Expression of serotonin (5-hydroxytryptamine, 5-HT) transporter mRNA in the rat brain was examined by in situ hybridization histochemistry with a synthetic oligonucleotide probe. 5-HT transporter mRNA was expressed in neurons in most of the raphe nuclei. The dorsal and median raphe nuclei contained intensely labeled neurons, while the caudal linear nucleus, raphe magnus nucleus, raphe pontis nucleus, raphe pallidus nucleus and the raphe obscurus nucleus contained weakly or moderately labeled neurons. The localization pattern of the 5-HT transporter mRNA-positive neurons coincides fairly well with that of 5-HT-immunoreactive neurons, indicating that 5-HT transporter is primarily located in serotonergic neurons.

Differential immunocytochemical localization of GABAA receptor gamma 1 and gamma 2 subunits in the rat brain.

Subunit-specific polyclonal antisera against the GABAA receptor gamma 1 and gamma 2 subunits were raised in rabbits and used for immunoblotting and immunocytochemistry of the rat brain. Each subunit protein was differentially distributed even in the region like cerebellar cortex where mRNAs of both subunits were distributed in the same manner. This may indicate that GABAA receptor gamma 1 and gamma 2 subunit proteins are subject to a subunit-specific subcellular sorting mechanism.

[Effects of exercise on immune functions in mice].

This study was carried out to examine the effects of forced running exercise on the immune functions of male ICR mice. Mice aged 4 weeks were divided into two groups: a non-exercise group (control) and a group given forced running exercise (exercise group). The exercise applied was forced running at 15m/min on a flat floor without any slope for 60 min a day. The duration of exercise was 5 days per week for 12 weeks. The results obtained were as follows: 1) After 12 weeks of forced running exercise, the weight of the anterior tibialis muscle and succinate dehydrogenase activity in the anterior tibialis muscle increased significantly (p < 0.01) in the exercise group compared with the control group. A tendency for thymus weight to increase was shown in the exercise group, and liver and spleen weights were significantly (p < 0.01) greater than in the control group. 2) The potentiation of phagocytic function of the reticulo-endothelial system, examined by the carbon clearance method, was seen in the exercise group. 3) The ability of peritoneal macrophages (M phi) to phagocytose latex beads significantly increased (p < 0.01) in the exercise group. The acid phosphatase activity of peritoneal M phi remained in both groups. However, lactate dehydrogenase activity of peritoneal M phi significantly increased (p < 0.01) in the exercise group compared with the control group. 4) The proliferation of splenocytes induced by Con A in the exercise group significantly increased (p < 0.01) compared with the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of oxytocin receptor messenger ribonucleic acid in the rat brain.

The expression of oxytocin receptor (OT-R) mRNA in the rat central nervous system was examined by in situ hybridization histochemistry using cRNA probe. Wide distribution of cells expressing OT-R mRNA was observed not only in the hypothalamus, but also in other regions. There were high levels of OT-R mRNA in the anterior olfactory nuclei, tenia tecta, olfactory tubercle, rostral most region of the frontal cortex, piriform cortex, layers 2 and 3 of the neocortex, bed nucleus of the stria terminalis, anterior medial preoptic nucleus (AV3V region), magnocellular preoptic nucleus, supraoptic nucleus, paraventricular hypothalamic nucleus, retrochiasmatic nucleus, ventromedial hypothalamic nucleus, paraventricular thalamic nucleus, central amygdaloid nucleus, medial amygdaloid nucleus, posterior cortical amygdaloid nucleus, amygdalohippocampal area, subiculum, prepositus hypoglossal nucleus, and dorsal motor nucleus of vagus. In most regions of the brain, our findings concurred with those obtained by receptor binding autoradiography using a ligand specific to OT. However, in the inferior olive nucleus, OT-R mRNA was not detected despite an abundance of binding sites showed by receptor binding autography. Despite this discrepancy OT appears to have central nervous system functions in addition to its hormonal functions.