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Food intake is regulated by a network of signals that emanate from the gut and the brainstem. The peripheral satiety signal cholecystokinin is released from the gut following food intake and acts on fibers of the vagus nerve, which project to the brainstem and activate neurons that modulate both gastrointestinal function and appetite. In this study, we found that neurons in the nucleus tractus solitarii of the brainstem that express prolactin-releasing peptide (PrRP) are activated rapidly by food ingestion. To further examine the role of this peptide in the control of food intake and energy metabolism, we generated PrRP-deficient mice and found that they displayed late-onset obesity and adiposity, phenotypes that reflected an increase in meal size, hyperphagia, and attenuated responses to the anorexigenic signals cholecystokinin and leptin. Hypothalamic expression of 6 other appetite-regulating peptides remained unchanged in the PrRP-deficient mice. Blockade of endogenous PrRP signaling in WT rats by central injection of PrRP-specific mAb resulted in an increase in food intake, as reflected by an increase in meal size. These data suggest that PrRP relays satiety signals within the brain and that selective disturbance of this system can result in obesity and associated metabolic disorders.

Chicken muscular dystrophy with abnormal muscle (AM) has been studied for more than 50 years, but the gene responsible for it remains unclear. Our previous studies narrowed down the AM candidate region to approximately 1Mbp of chicken chromosome 2q containing seven genes. In this study, we performed sequence comparison and gene expression analysis to elucidate the responsible gene. One missense mutation was detected in AM candidate genes, while no remarkable alteration of expression patterns was observed. The mutation was identified in WWP1, detected only in dystrophic chickens within several tetrapods. These results suggested WWP1 is responsible for chicken muscular dystrophy.

A positioning system based on a new zooming interferometer was developed for calibrating ultrahigh-resolution linear encoders. In the system, the positioning resolution is scaled down from that of the driving stage to that of the controlled stage by a zooming ratio that is determined by the ratio between the wavelengths of the optical sources for the zooming interferometer, so that the resolution in the controlled stage becomes very high. A femtosecond optical comb is used to stabilize two external-cavity diode lasers that are used as optical sources for the interferometer. The system realized a resolution of better than 30 pm and a stability of 1 nm.

Metastin/kisspeptin, the KiSS-1 gene product, has been identified as an endogenous ligand of GPR54 that reportedly regulates GnRH/LH surges and estrous cyclicity in female rats. The aim of the present study was to determine if metastin/kisspeptin neurons are a target of estrogen positive feedback to induce GnRH/LH surges. We demonstrated that preoptic area (POA) infusion of the anti-rat metastin/kisspeptin monoclonal antibody blocked the estrogen-induced LH surge, indicating that endogenous metastin/kisspeptin released around the POA mediates the estrogen positive feedback effect on GnRH/LH release. Metastin/kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) may be responsible for mediating the feedback effect because the percentage of c-Fos-expressing KiSS-1 mRNA-positive cells to total KiSS-1 mRNA-positive cells was significantly higher in the afternoon than in the morning in the anteroventral periventricular nucleus (AVPV) of high estradiol (E(2))-treated females. The percentage of c-Fos-expressing metastin/kisspeptin neurons was not different between the afternoon and morning in the arcuate nucleus (ARC). Most of the KiSS-1 mRNA expressing cells contain ERalpha immunoreactivity in the AVPV and ARC. In addition, AVPV KiSS-1 mRNA expressions were highest in the proestrous afternoon and lowest in the diestrus 1 in females and were increased by estrogen treatment in ovariectomized animals. On the other hand, the ARC KiSS-1 mRNA expressions were highest at diestrus 2 and lowest at proestrous afternoon and were increased by ovariectomy and decreased by high estrogen treatment. Males lacking the surge mode of GnRH/LH release showed no obvious cluster of metastin/kisspeptin-immunoreactive neurons in the AVPV when compared with high E(2)-treated females, which showed a much greater density of these neurons. Taken together, the present study demonstrates that the AVPV metastin/kisspeptin neurons are a target of estrogen positive feedback to induce GnRH/LH surges in female rats.

A technique for the measurement of the group refractive index wavelength dependence of optical materials using a low-coherence tandem interferometer and a spectrometer is proposed. Four channeled spectra resulting from interferences of light beams from different pairs of optical paths are used for the calculation of optical path differences. The group refractive index wavelength dependence is calculated from these optical path differences generated from the sample under measurement. No a priori information of the geometric thickness of a sample is required. The wavelength dependence of the group refractive index of the samples BK7 parallel plate of 5.200 and 10.025 mm from 675 to 850 nm is experimentally measured with an accuracy of the order of 10(-5) and a repeatability of the order of 10(-9).

A continuous-wave (cw) optical frequency synthesizer is demonstrated by using a monolithic-type cw optical parametric oscillator (cw-OPO) and an optical frequency comb. The cw-OPO is phase locked to an optical frequency comb that is phase locked to an atomic clock. The output frequency of the cw-OPO is frequency shifted with an electro-optic modulator, which makes it possible to tune the frequency continuously over 10 GHz. Furthermore, Doppler-free spectroscopy is performed using the optical frequency synthesizer for a cesium D1 line at 895 nm. The observed linewidth of 5 MHz is the natural linewidth of cesium. The center frequency of the line is consistent with a previous report.

Design, synthesis, and structure-activity relationships of thieno[2,3-b]pyridin-4-one-based non-peptide luteinizing hormone-releasing hormone (LHRH) receptor antagonists are described. Starting with the thienopyridin-4-one derivative 26d (T-98475) an optimization study was performed, which resulted in the identification of a highly potent and orally bioavailable LHRH receptor antagonist, 3-(N-benzyl-N-methylaminomethyl)-7-(2,6-difluorobenzyl)-4,7-dihydro-2-[4-(1-hydroxy-1-cyclopropanecarboxamido)phenyl]-5-isobutyryl-4-oxothieno[2,3-b]pyridine (33c). Compound 33c displayed subnanomolar in vitro activities for the human receptor and its oral administration caused effective suppression of the plasma LH levels in castrated male cynomolgus monkeys. Furthermore, SAR studies revealed that a hydroxyalkylamido moiety on the 2-phenyl ring is virtually equivalent to an alkylureido moiety, at least in this series of compounds.

We have developed a fiber-based frequency comb system consisting of a simple mode-locked fiber laser and a backward pumping amplifier combined with a highly nonlinear fiber with a short zerodispersion wavelength. As a result, the signal to noise ratio of the obtained carrier-envelope-offset frequency beat is larger than 45 dB at a bandwidth of 100 kHz. Furthermore, we have succeeded in measuring the optical frequencies of a 1542-nm acetylene-stabilized laser and a 532-nm iodinestabilized Nd:YAG laser continuously for more than one week using the fiber-based comb system. The long-term measurement revealed that the frequency stability of the iodine-stabilized laser was 5.7 x 10(-15) with 100 000 s averaging.

Ovulation is caused by a sequence of neuroendocrine events: GnRH and LH surges that are induced by positive feedback action of estrogen secreted by the mature ovarian follicles. The central mechanism of positive feedback action of estrogen on GnRH/LH secretion, however, is not fully understood yet. The present study examined whether metastin, the product of metastasis suppressor gene KiSS-1, is a central neuropeptide regulating GnRH/LH surge and then estrous cyclicity in the female rat. Metastin had a profound stimulation on LH secretion by acting on the preoptic area (POA), where most GnRH neurons projecting to the median eminence are located, because injection of metastin into the third ventricle or POA increased plasma LH concentrations in estrogen-primed ovariectomized rats. Metastin neurons were immunohistochemically found in the arcuate nucleus (ARC) to be colocalized with estrogen receptors with some fibers in the preoptic area (POA) in close apposition with GnRH neuronal cell bodies or fibers. Quantitative RT-PCR has revealed that KiSS-1 and GPR54 mRNAs were expressed in the ARC and POA, respectively. The blockade of local metastin action in the POA with a specific monoclonal antibody to rat metastin completely abolished proestrous LH surge and inhibited estrous cyclicity. Metastin-immunoreactive cell bodies in the ARC showed a marked increase and c-Fos expression in the early proestrus afternoon compared with the day of diestrus. Thus, metastin released in the POA is involved in inducing the preovulatory LH surge and regulating estrous cyclicity.

We have established a frequency measurement system for frequency-stabilized lasers operating in telecommunication wavelength bands, by using a femtosecond optical comb without the need for carrier envelope offset frequency control. This system has been used to measure the frequency of an acetylene-stabilized laser operating at 1542 nm for a period of over 10 hours. The frequency stability of the acetylene-stabilized laser is estimated to be 3x10-12 for a 10-s averaging time, improving toward 1x10-13 after 10000 s. We have measured three acetylene-stabilized lasers, including one commercially available laser, and confirmed that the frequency values are in good agreement (a frequency scatter of 2.1 kHz) with previously measured results reported by different institutes. In addition to the P(16) line of acetylene at 1542 nm, we measured the absolute frequencies of the P(24) line at 1547 nm, the P(1) line at 1534 nm, and the R(5) line at 1530 nm with a view to improving the accuracy of the acetylene frequency atlas. The acetylene-stabilized laser serves as an important optical frequency standard for telecommunication applications.

Prolactin-releasing peptide (PrRP) is a novel peptide found in bovine hypothalamus as an endogenous ligand of an orphan G-protein-coupled receptor (hGR3). It is known that PrRP is widely distributed and plays roles in the central nervous system (CNS). In particular, PrRP acts as a neurotransmitter that mediates stress and activates the hypothalamo-pituitary-adrenal axis. On the other hand, only a few studies have so far been performed on PrRP in peripheral tissues. Among peripheral tissues, appreciable levels of PrRP are found only in the adrenal gland; however, the PrRP-producing cells in the adrenal gland have not been identified. In this study, we detected PrRP mRNA in the rat adrenal medulla. So, we tried to identify the PrRP-producing cells in primary culture cells of the adrenal medulla. We found immunopositive PrRP cells among the cultured cells from the adrenal gland, but not in the adrenal gland tissue, by means of immunocytochemistry. The PrRP immunopositive cells were double positive for tyrosine hydroxylase (TH) and for phenylethanolamine N-methyltransferase (PNMT), which indicates that PrRP may be produced in a part of the adrenaline cells in the adrenal gland. This is the first report that PrRP is produced in the adrenaline-containing cells of the adrenal gland.

We have clarified the influences of water-vapor absorption lines in air on femtosecond pulse propagations from experimental and theoretical points of view. Precise measurements for the femtosecond pulse shapes after propagation of as much as 300 m through air have been made in a semiunderground optical testing tunnel. We observed the pulse splitting and the enhancement of the pulse broadening due to the 100-m propagation in air. The experimental results are in good agreement with the theoretical analysis by use of the HITRAN database at the edge and the center of the water-vapor absorption regions in air. Measured autocorrelation traces are mostly reproduced by those calculated with only the real part (dispersion) of the refractive index modulated by water-vapor absorption lines in air. This fact enables us to conclude that the pulse deformations due to water absorption lines in air are caused mainly by the real part (dispersion), not by the imaginary part (absorption), of the refractive index of air.

A Fourier transform spectrometer with heterodyne modulation achieved by a moving diffraction grating has been developed for the near-infrared (NIR) region. The grating simultaneously acts as a beam splitter and a modulator, which realizes the optical frequency shift of incident light for increasing the sensitivity of measurements by the heterodyne detection technique. The differences in diffraction angle among broad spectra are compensated by a collimating mirror and plane mirrors. The proposed spectrometer is used for the measurements of spectra in the NIR region. The signal-to-noise ratio of measurements is improved sevenfold with a heterodyne modulation of 410 Hz. As examples, this spectrometer is applied for quantitative calibration and discrimination of organic solutions. The measurement of transmission spectra of a grape is also demonstrated.

Galanin-like peptide (GALP) is a novel peptide that has been isolated from the porcine hypothalamus. The expression of GALP mRNA is localized to the hypothalamic arcuate nucleus and is thought to be under the regulation of leptin. First, we confirmed by real-time PCR analysis that sc administration of leptin to Wistar rats under food-deprived conditions resulted in a 1.5-fold increase in hypothalamic GALP mRNA levels. Next, GALP mRNA levels were found to be reduced by 50% in 11-wk-old male Zucker obese rats compared with age-matched Zucker lean rats, whereas neuropeptide Y mRNA levels were increased by 55% and proopiomelanocortin mRNA levels were reduced by 53% in Zucker obese rats. Analysis using a two-site enzyme immunoassay revealed a lower level of hypothalamic GALP immunoreactivity in 11-wk-old Zucker obese rats (5.9 fmol/mg protein) than in age-matched Zucker lean rats (19.6 fmol/mg protein). Immunohistochemical studies demonstrated that Zucker obese rats (11 wk old) had a reduced number of GALP immunoreactivity-positive cells (29.4 cells/3 slices) in the arcuate nucleus compared with age-matched Zucker lean rats (115 cells/3 slices). Furthermore, Zucker obese rats showed increased sensitivity to intracerebroventricularly administered GALP compared with Zucker lean rats, in that a lower dose of GALP increased plasma LH levels in male Zucker obese rats, but not in male Zucker lean rats. In addition, a reduction in the level of hypothalamic GALP mRNA was found in db/db and ob/ob mice. The result supports the hypothesis that the hypothalamic GALP gene expression is controlled by leptin signals and suggests possible involvement of GALP in the reproductive abnormalities of the Zucker obese rat.

Metastin is a novel peptide that was recently isolated from human placenta as the endogenous ligand of an orphan heptahelical receptor, hOT7T175. Metastin has been shown to suppress the motility of hOT7T175-transfected melanoma cells; however, studies of the physiological function of metastin have begun only recently. To investigate the possibility that metastin is an endocrine peptide, we determined the immunoreactive (ir-) metastin concentration in human plasma using our newly developed, sensitive, and specific two-site enzyme immunoassay. The plasma concentrations of ir-metastin in males and females were 1.30 +/- 0.14 (n = 12) and 1.31 +/- 0.37 fmol/ml (n = 10), respectively. As metastin is known to be abundant in human placenta, the ir-metastin concentration in the maternal plasma was then determined. The ir-metastin concentrations were 1230 +/- 346 fmol/ml (n = 11) in the first trimester, 4590 +/- 555 (n = 16) in the second trimester, and 9590 +/- 1640 (n = 12) in the third trimester. On d 5 after delivery, the ir-metastin concentration returned to nearly the nonpregnant level (7.63 +/- 1.33 fmol/ml; n = 10), suggesting that ir-metastin increases in pregnancy and is derived mainly from the placenta. The plasma from both nonpregnant and pregnant women showed a single ir-metastin peak at the same retention time as authentic metastin on reverse phase HPLC analysis, indicating that the major portion of the circulating metastin, as determined by our two-site enzyme immunoassay, represents endogenous metastin. Histochemical studies of human placenta localized metastin mRNA and immunoreactivity to the syncytiotrophoblasts. The present study provides evidence for metastin as a novel placenta-derived hormone in humans.

A four-year-old male cat was presented with regurgitation. Thoracic radiography and contrast radiogram showed a large oval mass and elevated esophagus. Exploratory thoracotomy showed omental herniation into the posterior mediastium through the esophageal hiatus. Because the mass of the omental herniation was so large, celiotomy through a paracostal incision was combined in order to return the omentum to its normal position. The diameter of the esophageal hiatus was approximately 1 cm but no fibrous adhesion was observed around the hiatus. Continuous 1-0 surgical sutures on the hiatus reduced the diameter of the hiatus. The cat made a successful postoperative recovery without complications.

We demonstrate microscopic time-resolved two-dimensional (2D) imaging that is based on a femtosecond amplifying optical Kerr gate (fs-amp OKG). The contribution of the optical nonlinear effects to the transverse imaging performance and the limit of the transverse resolving power are investigated. The optical Kerr effect in the excited state with amplification, used in the fs-amp OKG, does not deteriorate the quality of the time-resolved image at transverse resolutions up to at least 5.5 microm. We obtain a femtosecond-time-resolved 2D image of a microscopic object with a transverse resolution of 1.7 microm.

The group refractive index of air in laboratory conditions is measured directly between adjacent femtosecond laser pulses by a new interferometry technique. Measurement of the repetition rate of the mode-locked pulse train that gives the maximum amplitude of the interference-signal envelope enables us to determine the group refractive index of air within a standard deviation of 2 x 10(-7). This simple method without vacuum reference is attractive for measuring the group refractive index needed for precise distance measurements in open fields.