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Makoto Izumiya Miyu Haniu Katsuya Ueda Haruka Ishida Chuang Ma Hirokazu Ideta Atsushi Sobajima Koki Ueshiba Takeshi Uemura Naoto Saito Hisao Haniu 《International journal of molecular sciences》2021,22(14)
Many biomaterials have been evaluated using cultured cells. In particular, osteoblast-like cells are often used to evaluate the osteocompatibility, hard-tissue-regeneration, osteoconductive, and osteoinductive characteristics of biomaterials. However, the evaluation of biomaterial osteogenesis-inducing capacity using osteoblast-like cells is not standardized; instead, it is performed under laboratory-specific culture conditions with different culture media. However, the effect of different media conditions on bone formation has not been investigated. Here, we aimed to evaluate the osteogenesis of MC3T3-E1 cells, one of the most commonly used osteoblast-like cell lines for osteogenesis evaluation, and assayed cell proliferation, alkaline phosphatase activity, expression of osteoblast markers, and calcification under varying culture media conditions. Furthermore, the various media conditions were tested in uncoated plates and plates coated with collagen type I and poly-L-lysine, highly biocompatible molecules commonly used as pseudobiomaterials. We found that the type of base medium, the presence or absence of vitamin C, and the freshness of the medium may affect biomaterial regeneration. We posit that an in vitro model that recapitulates in vivo bone formation should be established before evaluating biomaterials. 相似文献
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Shotaro Honda Chiaki Murakami Haruka Yamada Yuki Murakami Ayuka Ishizaki Fumio Sakane 《Lipids》2019,54(11-12):763-771
Specific inhibitors of diacylglycerol kinase (DGK) ζ can be promising anticancer medications via the activation of cancer immunity. Although the detection of cellular activities of target enzymes is essential for drug screening in addition to in vitro assays, it is difficult to detect the activity of DGKζ in cells. In the present study, we generated AcGFP-DGKζ cDNA with a consensus N-myristoylation sequence at the 5′ end (Myr-AcGFP-DGKζ) to target DGKζ to membranes. Using liquid chromatography (LC)-tandem mass spectrometry (MS/MS) (LC–MS/MS), we showed that Myr-AcGFP-DGKζ, but not AcGFP-DGKζ without the myristoylation sequence, substantially augmented the levels of several phosphatidic acid (PtdOH) species. In contrast to Myr-AcGFP-DGKζ, its inactive mutant did not exhibit an increase in PtdOH production, indicating that the increase in PtdOH production was DGK activity-dependent. This method will be useful in chemical compound selection for the development of drugs targeting DGKζ and can be applicable to various soluble (nonmembrane bound) lipid-metabolizing enzymes, including other DGK isozymes. 相似文献
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Yukino Ogura Kazuko Tajiri Nobuyuki Murakoshi DongZhu Xu Saori Yonebayashi Siqi Li Yuta Okabe Duo Feng Yuzuno Shimoda Zoughu Song Haruka Mori Zixun Yuan Kazutaka Aonuma Masaki Ieda 《International journal of molecular sciences》2021,22(2)
Neutrophils are recruited into the heart at an early stage following a myocardial infarction (MI). These secrete several proteases, one of them being neutrophil elastase (NE), which promotes inflammatory responses in several disease models. It has been shown that there is an increase in NE activity in patients with MI; however, the role of NE in MI remains unclear. Therefore, the present study aimed to investigate the role of NE in the pathogenesis of MI in mice. NE expression peaked on day 1 in the infarcted hearts. In addition, NE deficiency improved survival and cardiac function post-MI, limiting fibrosis in the noninfarcted myocardium. Sivelestat, an NE inhibitor, also improved survival and cardiac function post-MI. Flow cytometric analysis showed that the numbers of heart-infiltrating neutrophils and inflammatory macrophages (CD11b+F4/80+CD206low cells) were significantly lower in NE-deficient mice than in wild-type (WT) mice. At the border zone between intact and necrotic areas, the number of terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive apoptotic cells was lower in NE-deficient mice than in WT mice. Western blot analyses revealed that the expression levels of insulin receptor substrate 1 and phosphorylation of Akt were significantly upregulated in NE-knockout mouse hearts, indicating that NE deficiency might improve cardiac survival by upregulating insulin/Akt signaling post-MI. Thus, NE may enhance myocardial injury by inducing an excessive inflammatory response and suppressing Akt signaling in cardiomyocytes. Inhibition of NE might serve as a novel therapeutic target in the treatment of MI. 相似文献
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Dr. Haruka Hiraoka Dr. Zhaoma Shu Dr. Bao Tri Le Dr. Keiko Masuda Dr. Kosuke Nakamoto Lyu Fangjie Dr. Naoko Abe Dr. Fumitaka Hashiya Dr. Yasuaki Kimura Dr. Yoshihiro Shimizu Prof. Dr. Rakesh N. Veedu Prof. Dr. Hiroshi Abe 《Chembiochem : a European journal of chemical biology》2021,22(24):3437-3442
We have found that antisense oligonucleotides and siRNA molecules modified with repeat structures of disulfide units can be directly introduced into the cytoplasm and exhibit a suppressive effect on gene expression. In this study, we analyzed the mechanism of cellular uptake of these membrane-permeable oligonucleotides (MPONs). Time-course analysis by confocal microscopy showed that the uptake of MPONs from the plasma membrane to the cytoplasm reached 50 % of the total uptake in about 5 min. In addition, analysis of the plasma membrane proteins to which MPONs bind, identified several proteins, including voltage-dependent anion channel. Next, we analyzed the behavior of MPONs in the cell and found them to be abundant in the nucleus as early as 24 h after addition with the amount increasing further after 48 and 72 h. The amount of MPONs was 2.5-fold higher than that of unmodified oligonucleotides in the nucleus after 72 h. We also designed antisense oligonucleotides and evaluated the effect of MPONs on mRNA exon skipping using DMD model cells; MPONs caused exon skipping with 69 % efficiency after 72 h, which was three times higher than the rate of the control. In summary, the high capacity for intracytoplasmic and nuclear translocation of MPONs is expected to be useful for therapeutic strategies targeting exon skipping. 相似文献
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Masaki Kaneniwa Song Miao Chunhong Yuan Haruka Lida Yutaka Fukuda 《Journal of the American Oil Chemists' Society》2000,77(8):825-831
The lipid and fatty acid composition of muscle of 10 species of freshwater fish obtained from a market of Shanghai City was
examined. Total lipids (TL) ranged over 0.9–4.7% of muscle for all samples. The content of triacylglycerol (TG) in muscle
ranged over 0.2–3.4% and that of polar lipids (PL) was 0.5–1.3%. Differences of TL content were dependent on TG contents.
The predominant important fatty acids (>10% of the total fatty acids in TL) were 16∶0 and 18∶1n−9 with some 16∶1n−7, 18∶2n−6,
and 22∶6n−3. The polyunsaturated fatty acids (PUFA) content was 10.2–43.4%, and especially Chinese sea bass contained above
20% of 22∶6n−3 in the total fatty acids. There were higher levels of PUFA such as 20∶5n−3 and 22∶6n−3 in PL than in neutral
lipids. Muscle of the silver carp was stored at 20°C, and changes of lipid classes during storage were examined. Free fatty
acids increased, and PL decreased during storage. This phenomenon was inhibited by heating the muscle, suggesting that lipid
hydrolysis by phospholipase occurred in silver carp muscle. 相似文献
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Haruka Matsukura Hitoshi Yoshida Takamichi Nakamoto Hiroshi Ishida 《Journal of Mechanical Science and Technology》2010,24(1):253-256
This paper reports on an olfactory display system that enables the synchronized presentation of odors with airflow in a virtual
reality environment. In the proposed system, computational fluid dynamics simulation is conducted to calculate how an odor
released from its source is spread in a given airflow field. An odor vapor with an appropriate concentration is generated
and delivered to the user’s nose. A sensation that an odor is coming from a specific object is created even though the object
per se is not releasing the odor. Experimental results are presented to show the potential of the proposed system. 相似文献
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Takashi Takizawa Noboru Nakayama Hisao Haniu Kaoru Aoki Masanori Okamoto Hiroki Nomura Manabu Tanaka Atsushi Sobajima Kazushige Yoshida Takayuki Kamanaka Kumiko Ajima Ayumu Oishi Chika Kuroda Haruka Ishida Satomi Okano Shinsuke Kobayashi Hiroyuki Kato Naoto Saito 《Advanced materials (Deerfield Beach, Fla.)》2018,30(4)
Titanium plates are widely used in clinical settings because of their high bone affinity. However, owing to their high elastic modulus, these plates are not suitable for bone repair since their proximity to the bone surface for prolonged periods can cause stress shielding, leading to bone embrittlement. In contrast, titanium fiber plates prepared by molding titanium fibers into plates by simultaneously applying compression and shear stress at normal room temperature can have an elastic modulus similar to that of bone cortex, and stress shielding will not occur even when the plate lies flush against the bone's surface. Titanium fibers can form a porous structure suitable for cell adhesion and as a bone repair scaffold. A titanium fiber plate is combined with osteoblasts and shown that the titanium fiber plate is better able to facilitate bone tissue repair than the conventional titanium plate when implanted in rat bone defects. Capable of being used in close contact with bone for a long time, and even capable of promoting bone repair, titanium fiber plates have a wide range of applications, and are expected to make great contributions to clinical management of increasing bone diseases, including bone fracture repair and bone regenerative medicine. 相似文献