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Chondroitin Sulfate Synthase-2 Is Necessary for Chain Extension of Chondroitin Sulfate but Not Critical for Skeletal Development
Hiroyasu Ogawa, Sonoko Hatano, Nobuo Sugiura, Naoko Nagai, Takashi Sato, Katsuji Shimizu, Koji Kimata, Hisashi Narimatsu, Hideto Watanabe
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0043806
Abstract: Chondroitin sulfate (CS) is a linear polysaccharide consisting of repeating disaccharide units of N-acetyl-D-galactosamine and D-glucuronic acid residues, modified with sulfated residues at various positions. Based on its structural diversity in chain length and sulfation patterns, CS provides specific biological functions in cell adhesion, morphogenesis, neural network formation, and cell division. To date, six glycosyltransferases are known to be involved in the biosynthesis of chondroitin saccharide chains, and a hetero-oligomer complex of chondroitin sulfate synthase-1 (CSS1)/chondroitin synthase-1 and chondroitin sulfate synthase-2 (CSS2)/chondroitin polymerizing factor is known to have the strongest polymerizing activity. Here, we generated and analyzed CSS2?/? mice. Although they were viable and fertile, exhibiting no overt morphological abnormalities or osteoarthritis, their cartilage contained CS chains with a shorter length and at a similar number to wild type. Further analysis using CSS2?/? chondrocyte culture systems, together with siRNA of CSS1, revealed the presence of two CS chain species in length, suggesting two steps of CS chain polymerization; i.e., elongation from the linkage region up to Mr ~10,000, and further extension. There, CSS2 mainly participated in the extension, whereas CSS1 participated in both the extension and the initiation. Our study demonstrates the distinct function of CSS1 and CSS2, providing a clue in the elucidation of the mechanism of CS biosynthesis.
Validation of the japanese version of the sarcoidosis health questionnaire: A cross-sectional study
Kiminobu Tanizawa, Tomohiro Handa, Sonoko Nagai, Toru Oga, Takeshi Kubo, Yutaka Ito, Kizuku Watanabe, Kensaku Aihara, Kazuo Chin, Michiaki Mishima, Takateru Izumi
Health and Quality of Life Outcomes , 2011, DOI: 10.1186/1477-7525-9-34
Abstract: The SHQ was translated into Japanese following the forward-backward procedure. The reliability and validity of the Japanese version of the SHQ were examined. One hundred twenty-two Japanese patients with biopsy-proven sarcoidosis were evaluated by the SHQ, the Medical Outcomes Study 36-item short form (SF-36), the St. George's Respiratory Questionnaire (SGRQ), chest radiography, an electrocardiogram, laboratory blood tests, pulmonary function tests, an echocardiogram, and assessments of dyspnea and depressive symptoms. The SHQ was found to have acceptable levels of internal consistency (Cronbach's coefficient α values = 0.68 to 0.91). SHQ scores correlated significantly with scores on the SF-36 and SGRQ. The domain or total scores on the SHQ also significantly correlated with serum levels of the soluble interleukin-2 receptor, the percentage of the predicted forced vital capacity, pulmonary arterial systolic pressure, dyspnea, and depressive symptoms. Also, the SHQ scores of patients who had one or two organ systems affected by sarcoidosis were significantly different from those of patients who had three or more organ systems involvement.The Japanese version of the SHQ can be used to assess the HRQOL of patients with sarcoidosis.Sarcoidosis is a chronic and multisystem granulomatous disease of unknown etiology that can involve almost any organ system [1]. The assessment of overall disease burden is often challenging in sarcoidosis because of the disease's various clinical manifestations. Although spirometry and chest radiography are usually used to evaluate disease activity, these measures fail to address extrapulmonary lesions. As in other respiratory diseases, the patient's health-related quality of life (HRQOL) may be a useful measure to assess the systemic impact of sarcoidosis [2-4], and impaired HRQOL has been reported in sarcoidosis [5]. Given that sarcoidosis is a chronic disease for which there is no curative therapy yet [6], HRQOL could be the most importa
Clinical Relevance of Plasma Prostaglandin F2α Metabolite Concentrations in Patients with Idiopathic Pulmonary Fibrosis
Kensaku Aihara, Tomohiro Handa, Toru Oga, Kizuku Watanabe, Kiminobu Tanizawa, Kohei Ikezoe, Yoshio Taguchi, Hiroe Sato, Kazuo Chin, Sonoko Nagai, Shuh Narumiya, Athol U. Wells, Michiaki Mishima
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0066017
Abstract: Background Idiopathic pulmonary fibrosis (IPF) is a devastating lung disease of unknown etiology with few current treatment options. Recently, we determined an important role of prostaglandin F2α (PGF2α) in pulmonary fibrosis by using a bleomycin-induced pulmonary fibrosis model and found an abundance of PGF2α in bronchoalveolar lavage fluid of IPF patients. We investigated the role of PGF2α in human IPF by assessing plasma concentrations of 15-keto-dihydro PGF2α, a stable metabolite of PGF2α. Methods We measured plasma concentrations of 15-keto-dihydro PGF2α in 91 IPF patients and compared these values with those of controls (n = 25). We further investigated the relationships of plasma 15-keto-dihydro PGF2α concentrations with disease severity and mortality. Results Plasma concentrations of 15-keto-dihydro PGF2α were significantly higher in IPF patients than controls (p<0.001). Plasma concentrations of this metabolite were significantly correlated with forced expiratory volume in 1 second (Rs [correlation coefficient] = ?0.34, p = 0.004), forced vital capacity (Rs = ?0.33, p = 0.005), diffusing capacity for carbon monoxide (Rs = ?0.36, p = 0.003), the composite physiologic index (Rs = 0.40, p = 0.001), 6-minute walk distance (Rs = ?0.24, p = 0.04) and end-exercise oxygen saturation (Rs = ?0.25, p = 0.04) when patients with emphysema were excluded. Multivariate analysis using stepwise Cox proportional hazards model showed that a higher composite physiologic index (relative risk = 1.049, p = 0.002) and plasma 15-keto-dihydro PGF2α concentrations (relative risk = 1.005, p = 0.002) were independently associated with an increased risk of mortality. Conclusions We demonstrated significant associations of plasma concentrations of PGF2α metabolites with disease severity and prognosis, which support a potential pathogenic role for PGF2α in human IPF.
Characterization of Acid-Soluble Collagen from Skins of Surf Smelt (Hypomesus pretiosus japonicus Brevoort)  [PDF]
Takeshi Nagai
Food and Nutrition Sciences (FNS) , 2010, DOI: 10.4236/fns.2010.12010
Abstract: Acid-soluble collagen was extracted from the skins of surf smelt and characterized. The yield of collagen was high about 24.0% on a dry weight basis. By SDS-polyacrylamide gel electrophoresis and CM-Toyopearl 650 M column chromatography, this collagen is a heterotrimer with a chain composition of α1α2α3. The denaturation temperature was 32.5℃, about 4.5℃ lower than that from porcine skin. Attenuated total reflectance-fourier transform infrared analysis showed that the percentage of secondary structural components in this collagen were 11% α-helix, 34% β-sheet, 19% β-turn, and 21% others. It suggests that the triple helical structure is present in the acid-soluble collagen from the skins of the surf smelt in comparison to that from the skin of porcine.
Modified Cp Criterion for Optimizing Ridge and Smooth Parameters in the MGR Estimator for the Nonparametric GMANOVA Model  [PDF]
Isamu Nagai
Open Journal of Statistics (OJS) , 2011, DOI: 10.4236/ojs.2011.11001
Abstract: Longitudinal trends of observations can be estimated using the generalized multivariate analysis of variance (GMANOVA) model proposed by [10]. In the present paper, we consider estimating the trends nonparametrically using known basis functions. Then, as in nonparametric regression, an overfitting problem occurs. [13] showed that the GMANOVA model is equivalent to the varying coefficient model with non-longitudinal covariates. Hence, as in the case of the ordinary linear regression model, when the number of covariates becomes large, the estimator of the varying coefficient becomes unstable. In the present paper, we avoid the overfitting problem and the instability problem by applying the concept behind penalized smoothing spline regression and multivariate generalized ridge regression. In addition, we propose two criteria to optimize hyper parameters, namely, a smoothing parameter and ridge parameters. Finally, we compare the ordinary least square estimator and the new estimator.
Rapid progress of DNA replication studies in Archaea, the third domain of life
Yoshizumi Ishino,Sonoko Ishino
Science China Life Sciences , 2012, DOI: 10.1007/s11427-012-4324-9
Abstract: Archaea, the third domain of life, are interesting organisms to study from the aspects of molecular and evolutionary biology. Archaeal cells have a unicellular ultrastructure without a nucleus, resembling bacterial cells, but the proteins involved in genetic information processing pathways, including DNA replication, transcription, and translation, share strong similarities with those of Eukaryota. Therefore, archaea provide useful model systems to understand the more complex mechanisms of genetic information processing in eukaryotic cells. Moreover, the hyperthermophilic archaea provide very stable proteins, which are especially useful for the isolation of replisomal multicomplexes, to analyze their structures and functions. This review focuses on the history, current status, and future directions of archaeal DNA replication studies.
DNA polymerases as useful reagents for biotechnology – the history of developmental research in the field
Sonoko Ishino,Yoshizumi Ishino
Frontiers in Microbiology , 2014, DOI: 10.3389/fmicb.2014.00465
Abstract: DNA polymerase is a ubiquitous enzyme that synthesizes complementary DNA strands according to the template DNA in living cells. Multiple enzymes have been identified from each organism, and the shared functions of these enzymes have been investigated. In addition to their fundamental role in maintaining genome integrity during replication and repair, DNA polymerases are widely used for DNA manipulation in vitro, including DNA cloning, sequencing, labeling, mutagenesis, and other purposes. The fundamental ability of DNA polymerases to synthesize a deoxyribonucleotide chain is conserved. However, the more specific properties, including processivity, fidelity (synthesis accuracy), and substrate nucleotide selectivity, differ among the enzymes. The distinctive properties of each DNA polymerase may lead to the potential development of unique reagents, and therefore searching for novel DNA polymerase has been one of the major focuses in this research field. In addition, protein engineering techniques to create mutant or artificial DNA polymerases have been successfully developing powerful DNA polymerases, suitable for specific purposes among the many kinds of DNA manipulations. Thermostable DNA polymerases are especially important for PCR-related techniques in molecular biology. In this review, we summarize the history of the research on developing thermostable DNA polymerases as reagents for genetic manipulation and discuss the future of this research field.
Intralymphatic Histiocytosis with Rheumatoid Arthritis: Possible Association with the Joint Involvement  [PDF]
Takahide Kaneko, Sonoko Takeuchi, Hajime Nakano, Daisuke Sawamura
Case Reports in Clinical Medicine (CRCM) , 2014, DOI: 10.4236/crcm.2014.33036

Intralymphatic histiocytosis is characterized by dilated vessels containing aggregates of mononuclear histiocytes in their lumina, and the majority of cases have been associated with rheumatoid arthritis (RA). We report a case of a 61-year-old woman with RA who presented with livedo-like erythema and red papules around the right elbow and forearm. Histological examinations demonstrated numerous dilated vessels in the dermis partially filled with medium- to large-sized mononuclear cells. Immunohistochemically, most of the intraluminal cells were CD68-positive histiocytes. Endothelial cells were positive for D2-40, a marker for lymphatic endothelial cells. We diagnosed the patient with intralymphatic histiocytosis based on these clinical and histological findings. Interestingly, the skin manifestations exacerbated

Long-Lasting Consequences of Neonatal Maternal Separation on Social Behaviors in Ovariectomized Female Mice
Mumeko C. Tsuda, Sonoko Ogawa
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0033028
Abstract: Maternal separation (MS) stress is known to induce long-lasting alterations in emotional and anxiety-related behaviors, but effects on social behaviors are not well defined. The present study examined MS effects on female social behaviors in the social investigation (SIT) and social preference (SPT) tests, in addition to non-social behaviors in the open-field (OFT) and light-dark transition (LDT) tests in C57BL/6J mice. All females were tested as ovariectomized to eliminate confounding effects of endogenous estrogen during behavioral testing. Daily MS (3 hr) from postnatal day 1 to 14 did not affect anxiety levels in LDT, but were elevated in OFT with modified behavioral responses to the novel environment. Furthermore, MS altered social investigative behaviors and preference patterns toward unfamiliar stimulus mice in SIT and short- and long-term SPT paradigms. In SIT, MS reduced social investigation duration and increased number of stretched approaches towards both female and male unfamiliar stimulus mice, suggesting increased social anxiety levels in MS females. Similarly, MS heightened levels of social anxiety during short-term SPT but no MS effect on social preference was found. On the other hand, MS females displayed a distinctive preference for female stimuli, unlike control females, when tested for long-term SPT over a prolonged period of 5 days. Evaluation of FosB expression in the paraventricular nucleus, medial and central amygdala following stimulus exposure demonstrated greater number of FosB immunopositive cells in all three brain regions in MS females compared to control females. These results suggest that MS females might differ in neuroendocrine responses toward unfamiliar female and male opponents, which may be associated with modifications in social behaviors found in the present study. Taken together, this study provides new evidence that early life stress modifies female social behaviors by highlighting alterations in behavioral responses to situations involving social as well as non-social novelty.
Vertebral Bone Drilling (Puncture) Attenuates the Intractable Pain Due to Vertebral Fractures without Collapse  [PDF]
Koichi Ota, Hirosi Nagai
Open Journal of Anesthesiology (OJAnes) , 2016, DOI: 10.4236/ojanes.2016.64012
Abstract: Purpose: Osteoporotic vertebral fractures with no sign of vertebral collapse on initial radiographs, which is so-called occult vertebral fractures (VFs), exist. Occult VFs have a high rate of missed diagnosis, and the treatment of these fractures has rarely been discussed in the literature. We evaluated the effects of vertebral bone drilling for the pain due to occults VFs. Materials and Methods: Eighteen patients with painful osteoporotic occult VFs underwent the vertebral bone drilling. We evaluated the clinical outcome by comparing numerical rating scale (NRS) and activity of daily life (ADL) values between before and after the vertebral bone drilling. Comparisons were made by using Wilcoxon signed rank test. Results: The mean baseline NRS and ADL score, and the mean NRS and ALD score after the bone drilling were 8.4 ± 0.8, 2.2 ± 0.6, 2.4 ± 1.0, 4.6 ± 0.5, respectively. Among the patients, we detected significant improvements in NRS pain score and ADL score after the drilling compared with baseline score (p < 0.0002). Conclusion: Vertebral bone drilling at the site of painful vertebral compression fractures alleviated the intractable pain due to osteoporotic occult VFs.
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