リュウキュウマツ一斉枯死後にアカギが侵入した父島の二次林における31年間の動態

To quantify dynamics of a secondary forest that was invaded by Bischofia javanica, after death of a dominant tree, Pinus luchuensis, we represented long-term monitoring data of a 20 x 20 m plot in the Fukiagedani Valley in Chichijima Island for 31 years (1984-2015). With the death of P. luchuensis, basal area of trees of all woody species reduced to less than half before the death, and thereafter increased. The numbers of trees and saplings decreased during the period. This suggested that individual trees grew after death of P. luchuensis but recruitments of seedlings did not occur, which could be related to closure of forest canopy with the growth of individual trees. All data of the study was published on the web site of the Ogasawara Research Committee of Tokyo Metropolitan University ([email protected]).リュウキュウマツの一斉枯死後にアカギが侵入した森林の長期的な動態を父島の二次林における31 年間のデータから明らかにした。マツ枯れ直後、森林のBA は50%以下に減少した。その後31年間で全体のBAは増加し、マツ枯れ以前の約70%程度まで回復したと考えられる。樹木の個体数は、1990年まで増加し、その後減少した。これは、マツ枯れ後、樹木が成長し、現存量が増加する一方で、競争によって小径木の枯死と新規加入個体の減少を示唆する。この研究で使用したデータを公開する。利用を希望する場合の問い合わせ先は、以下のアドレス（[email protected]）である

Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe

The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA. The fluorescence intensity corresponding to the protected RNA fragment increased in a dose-dependent manner with respect to the complementary-strand RNA. More highly sensitive detection of miRNA by microchip electrophoresis than by conventional method using fluorescence-labeled riboprobe could be obtained in 180 s. An obvious increase in miR-222 expression induced by nerve growth factor in PC12 cells could be observed. These results clearly indicate the potential of microchip electrophoresis for the analysis of miRNA using RNase protection assay with a fluorescence-labeled riboprobe

Tumor Cell Detection among Leukocytes by Microchip

Background: Accurate detection and analysis of circulating tumor cells plays an important role in the diagnosis and treatment of metastatic cancer treatment. Methods and Findings: A cell microarray chip was used to detect spiked carcinoma cells among leukocytes. The chip, with 20,944 microchambers (105 µm width and 50 µm depth), was made from polystyrene; and the formation of monolayers of leukocytes in the microchambers was observed. Cultured human T lymphoblastoid leukemia (CCRF-CEM) cells were used to examine the potential of the cell microarray chip for the detection of spiked carcinoma cells. A T lymphoblastoid leukemia suspension was dispersed on the chip surface, followed by 15 min standing to allow the leukocytes to settle down into the microchambers. Approximately 29 leukocytes were found in each microchamber when about 600,000 leukocytes in total were dispersed onto a cell microarray chip. Similarly, when leukocytes isolated from human whole blood were used, approximately 89 leukocytes entered each microchamber when about 1,800,000 leukocytes in total were placed onto the cell microarray chip. After washing the chip surface, PE-labeled anti-cytokeratin monoclonal antibody and APC-labeled anti-CD326 (EpCAM) monoclonal antibody solution were dispersed onto the chip surface and allowed to react for 15 min; and then a microarray scanner was employed to detect any fluorescence-positive cells within 20 min. In the experiments using spiked carcinoma cells (NCI-H1650, 0.01 to 0.0001%), accurate detection of carcinoma cells was achieved with PE-labeled anti-cytokeratin monoclonal antibody. Furthermore, verification of carcinoma cells in the microchambers was performed by double staining with the above monoclonal antibodies. Conclusion: The potential application of the cell microarray chip for the detection of CTCs was shown, thus demonstrating accurate detection by double staining for cytokeratin and EpCAM at the single carcinoma cell level

Rapid and Highly Sensitive Detection of Malaria-Infected Erythrocytes Using a Cell Microarray Chip

BACKGROUND: Malaria is one of the major human infectious diseases in many endemic countries. For prevention of the spread of malaria, it is necessary to develop an early, sensitive, accurate and conventional diagnosis system. METHODS AND FINDINGS: A cell microarray chip was used to detect for malaria-infected erythrocytes. The chip, with 20,944 microchambers (105 µm width and 50 µm depth), was made from polystyrene, and the formation of monolayers of erythrocytes in the microchambers was observed. Cultured Plasmodium falciparum strain 3D7 was used to examine the potential of the cell microarray chip for malaria diagnosis. An erythrocyte suspension in a nuclear staining dye, SYTO 59, was dispersed on the chip surface, followed by 10 min standing to allow the erythrocytes to settle down into the microchambers. About 130 erythrocytes were accommodated in each microchamber, there being over 2,700,000 erythrocytes in total on a chip. A microarray scanner was employed to detect any fluorescence-positive erythrocytes within 5 min, and 0.0001% parasitemia could be detected. To examine the contamination by leukocytes of purified erythrocytes from human blood, 20 µl of whole blood was mixed with 10 ml of RPMI 1640, and the mixture was passed through a leukocyte isolation filter. The eluted portion was centrifuged at 1,000×g for 2 min, and the pellet was dispersed in 1.0 ml of medium. SYTO 59 was added to the erythrocyte suspension, followed by analysis on a cell microarray chip. Similar accommodation of cells in the microchambers was observed. The number of contaminating leukocytes was less than 1 on a cell microarray chip. CONCLUSION: The potential of the cell microarray chip for the detection of malaria-infected erythrocytes was shown, it offering 10-100 times higher sensitivity than that of conventional light microscopy and easy operation in 15 min with purified erythrocytes

Detection Chip for Malaria

Background: Malaria is one of the major human infectious diseases in many endemic countries. For prevention of the spread of malaria, it is necessary to develop an early, sensitive, accurate and conventional diagnosis system. Methods and Findings: A cell microarray chip was used to detect for malaria-infected erythrocytes. The chip, with 20,944 microchambers (105 µm width and 50 µm depth), was made from polystyrene, and the formation of monolayers of erythrocytes in the microchambers was observed. Cultured Plasmodium falciparum strain 3D7 was used to examine the potential of the cell microarray chip for malaria diagnosis. An erythrocyte suspension in a nuclear staining dye, SYTO 59, was dispersed on the chip surface, followed by 10 min standing to allow the erythrocytes to settle down into the microchambers. About 130 erythrocytes were accommodated in each microchamber, there being over 2,700,000 erythrocytes in total on a chip. A microarray scanner was employed to detect any fluorescence-positive erythrocytes within 5 min, and 0.0001% parasitemia could be detected. To examine the contamination by leukocytes of purified erythrocytes from human blood, 20 µl of whole blood was mixed with 10 ml of RPMI 1640, and the mixture was passed through a leukocyte isolation filter. The eluted portion was centrifuged at 1,000×g for 2 min, and the pellet was dispersed in 1.0 ml of medium. SYTO 59 was added to the erythrocyte suspension, followed by analysis on a cell microarray chip. Similar accommodation of cells in the microchambers was observed. The number of contaminating leukocytes was less than 1 on a cell microarray chip. Conclusion: The potential of the cell microarray chip for the detection of malaria-infected erythrocytes was shown, it offering 10–100 times higher sensitivity than that of conventional light microscopy and easy operation in 15 min with purified erythrocytes

Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome.

The loss-of-function mutations of serine protease inhibitor, Kazal type 1 (SPINK1) gene are associated with human chronic pancreatitis, but the underlying mechanisms remain unknown. We previously reported that mice lacking Spink3, the murine homologue of human SPINK1, die perinatally due to massive pancreatic acinar cell death, precluding investigation of the effects of SPINK1 deficiency. To circumvent perinatal lethality, we have developed a novel method to integrate human SPINK1 gene on the X chromosome using Cre-loxP technology and thus generated transgenic mice termed "X-SPINK1". Consistent with the fact that one of the two X chromosomes is randomly inactivated, X-SPINK1 mice exhibit mosaic pattern of SPINK1 expression. Crossing of X-SPINK1 mice with Spink3+/- mice rescued perinatal lethality, but the resulting Spink3-/-;XXSPINK1 mice developed spontaneous pancreatitis characterized by chronic inflammation and fibrosis. The results show that mice lacking a gene essential for cell survival can be rescued by expressing this gene on the X chromosome. The Spink3-/-;XXSPINK1 mice, in which this method has been applied to partially restore SPINK1 function, present a novel genetic model of chronic pancreatitis

Calibration of the AKARI Far-Infrared Imaging Fourier Transform Spectrometer

The Far-Infrared Surveyor (FIS) onboard the AKARI satellite has a spectroscopic capability provided by a Fourier transform spectrometer (FIS-FTS). FIS-FTS is the first space-borne imaging FTS dedicated to far-infrared astronomical observations. We describe the calibration process of the FIS-FTS and discuss its accuracy and reliability. The calibration is based on the observational data of bright astronomical sources as well as two instrumental sources. We have compared the FIS-FTS spectra with the spectra obtained from the Long Wavelength Spectrometer (LWS) of the Infrared Space Observatory (ISO) having a similar spectral coverage. The present calibration method accurately reproduces the spectra of several solar system objects having a reliable spectral model. Under this condition the relative uncertainty of the calibration of the continuum is estimated to be $\pm$ 15% for SW, $\pm$ 10% for 70-85 cm^(-1) of LW, and $\pm$ 20% for 60-70 cm^(-1) of LW; and the absolute uncertainty is estimated to be +35/-55% for SW, +35/-55% for 70-85 cm^(-1) of LW, and +40/-60% for 60-70 cm^(-1) of LW. These values are confirmed by comparison with theoretical models and previous observations by the ISO/LWS.Comment: 22 pages, 10 figure

Calibration and Performance of the AKARI Far-Infrared Surveyor (FIS) -- Slow-Scan Observation Mode for Point Sources

We present the characterization and calibration of the Slow-Scan observation mode of the Far-Infrared Surveyor (FIS) onboard the AKARI satellite. The FIS, one of the two focal-plane instruments on AKARI, has four photometric bands between 50--180 um with two types of Ge:Ga array detectors. In addition to the All-Sky Survey, FIS has also taken detailed far-infrared images of selected targets by using the Slow-Scan mode. The sensitivity of the Slow-Scan mode is one to two orders of magnitude better than that of the All-Sky Survey, because the exposure time on a targeted source is much longer. The point spread functions (PSFs) were obtained by observing several bright point-like objects such as asteroids, stars, and galaxies. The derived full widths at the half maximum (FWHMs) are ~30'' for the two shorter wavelength bands and ~40'' for the two longer wavelength bands, being consistent with those expected by the optical simulation, although a certain amount of excess is seen in the tails of the PSFs. The flux calibration has been performed by the observations of well-established photometric calibration standards (asteroids and stars) in a wide range of fluxes. After establishing the method of aperture photometry, the photometric accuracy for point-sources is better than +-15% in all of the bands expect for the longest wavelength.Comment: 25 pages, 7 figures, accepted publication in PAS

The Far-Infrared Surveyor (FIS) for AKARI

The Far-Infrared Surveyor (FIS) is one of two focal plane instruments on the AKARI satellite. FIS has four photometric bands at 65, 90, 140, and 160 um, and uses two kinds of array detectors. The FIS arrays and optics are designed to sweep the sky with high spatial resolution and redundancy. The actual scan width is more than eight arcmin, and the pixel pitch is matches the diffraction limit of the telescope. Derived point spread functions (PSFs) from observations of asteroids are similar to the optical model. Significant excesses, however, are clearly seen around tails of the PSFs, whose contributions are about 30% of the total power. All FIS functions are operating well in orbit, and its performance meets the laboratory characterizations, except for the two longer wavelength bands, which are not performing as well as characterized. Furthermore, the FIS has a spectroscopic capability using a Fourier transform spectrometer (FTS). Because the FTS takes advantage of the optics and detectors of the photometer, it can simultaneously make a spectral map. This paper summarizes the in-flight technical and operational performance of the FIS.Comment: 23 pages, 10 figures, and 2 tables. Accepted for publication in the AKARI special issue of the Publications of the Astronomical Society of Japa

Ancestral TSH mechanism signals summer in a photoperiodic mammal

SummaryIn mammals, day-length-sensitive (photoperiodic) seasonal breeding cycles depend on the pineal hormone melatonin, which modulates secretion of reproductive hormones by the anterior pituitary gland [1]. It is thought that melatonin acts in the hypothalamus to control reproduction through the release of neurosecretory signals into the pituitary portal blood supply, where they act on pituitary endocrine cells [2]. Contrastingly, we show here that during the reproductive response of Soay sheep exposed to summer day lengths, the reverse applies: Melatonin acts directly on anterior-pituitary cells, and these then relay the photoperiodic message back into the hypothalamus to control neuroendocrine output. The switch to long days causes melatonin-responsive cells in the pars tuberalis (PT) of the anterior pituitary to increase production of thyrotrophin (TSH). This acts locally on TSH-receptor-expressing cells in the adjacent mediobasal hypothalamus, leading to increased expression of type II thyroid hormone deiodinase (DIO2). DIO2 initiates the summer response by increasing hypothalamic tri-iodothyronine (T3) levels. These data and recent findings in quail [3] indicate that the TSH-expressing cells of the PT play an ancestral role in seasonal reproductive control in vertebrates. In mammals this provides the missing link between the pineal melatonin signal and thyroid-dependent seasonal biology