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61.
62.
Takuma Inai Tomoya Takabayashi Mutsuaki Edama Masayoshi Kubo 《Biomedical engineering online》2018,17(1):177
Background
Sit-to-stand movements are a necessary part of daily life, and excessive mechanical stress on the articular cartilage has been reported to encourage the progression of osteoarthritis. Although a change in hip joint angle at seat-off may affect hip joint contact force during a sit-to-stand movement, the effect is unclear. This study aimed to examine the effect of the hip joint angle at seat-off on the hip joint contact force during a sit-to-stand movement by using a computer simulation.Methods
A musculoskeletal model was created for the computer simulation, and eight muscles were attached to each lower limb. Various sit-to-stand movements were generated using parameters (e.g., seat height and time from seat-off to standing posture) reported by previous studies. The hip joint contact force for each sit-to-stand movement was calculated. Furthermore, the effect of the hip joint angle at seat-off on the hip joint contact force during the sit-to-stand movement was examined. In this study, as the changes to the musculoskeletal model parameters affect the hip joint contact force, a sensitivity analysis was conducted.Results and conclusions
The hip joint contact force during the sit-to-stand movement increased approximately linearly as the hip flexion angle at the seat-off increased. Moreover, the normal sit-to-stand movement and the sit-to-stand movement yielding a minimum hip joint contact force were approximately equivalent. The effect of the changes to the musculoskeletal model parameters on the main findings of this study was minimal. Thus, the main findings are robust and may help prevent the progression of hip osteoarthritis by decreasing mechanical stress, which will be explored in future studies.63.
Natsumi Takei Takuma Nakamura Shohei Kawamura Yuki Takada Yui Satoh Atsushi P. Kimura Tomoya Kotani 《Biological procedures online》2018,20(1):6
Background
Subcellular localization of coding and non-coding RNAs has emerged as major regulatory mechanisms of gene expression in various cell types and many organisms. However, techniques that enable detection of the subcellular distribution of these RNAs with high sensitivity and high resolution remain limited, particularly in vertebrate adult tissues and organs. In this study, we examined the expression and localization of mRNAs encoding Pou5f1/Oct4, Mos, Cyclin B1 and Deleted in Azoospermia-like (Dazl) in zebrafish and mouse ovaries by combining tyramide signal amplification (TSA)-based in situ hybridization with paraffin sections which can preserve cell morphology of tissues and organs at subcellular levels. In addition, the distribution of a long non-coding RNA (lncRNA), lncRNA-HSVIII, in mouse testes was examined by the same method.Results
The mRNAs encoding Mos, Cyclin B1 and Dazl were found to assemble into distinct granules that were distributed in different subcellular regions of zebrafish and mouse oocytes, suggesting conserved and specific regulations of these mRNAs. The lncRNA-HSVIII was first detected in the nucleus of spermatocytes at prophase I of the meiotic cell cycle and was then found in the cytoplasm of round spermatids, revealing expression patterns of lncRNA during germ cell development. Collectively, the in situ hybridization method demonstrated in this study achieved the detection and comparison of precise distribution patterns of coding and non-coding RNAs at subcellular levels in single cells of adult tissues and organs.Conclusions
This high-sensitivity and high-resolution in situ hybridization is applicable to many vertebrate species and to various tissues and organs and will be useful for studies on the subcellular regulation of gene expression at the level of RNA localization.64.
65.
Takuya Kawanai Yukio Ago Ryo Watanabe Aya Inoue Atsuki Taruta Yusuke Onaka Shigeru Hasebe Hitoshi Hashimoto Toshio Matsuda Kazuhiro Takuma 《Neurochemical research》2016,41(10):2574-2584
Valproic acid (VPA) is a multi-target drug and an inhibitor of histone deacetylase (HDAC). We have previously demonstrated that prenatal exposure to VPA at embryonic day 12.5 (E12.5), but not at E14.5, causes autism-like behavioral abnormalities in male mouse offspring. We have also found that prenatal VPA exposure causes transient histone hyperacetylation in the embryonic brain, followed by decreased neuronal cell numbers in the prefrontal and somatosensory cortices after birth. In the present study, we examined whether prenatal HDAC inhibition affects neuronal maturation in primary mouse cortical neurons. Pregnant mice were injected intraperitoneally with VPA (500 mg/kg) and the more selective HDAC inhibitor trichostatin A (TSA; 500 µg/kg) at E12.5 or E14.5, and primary neuronal cultures were prepared from the cerebral cortices of their embryos. Prenatal exposure to VPA at E12.5, but not at E14.5, decreased total number, total length, and complexity of neuronal dendrites at 14 days in vitro (DIV). The effects of VPA weakened at 21 DIV. Exposure to TSA at E12.5, but not at E14.5, also delayed maturation of cortical neurons. In addition, real-time quantitative PCR revealed that the prenatal exposure to TSA decreased neuroligin-1 (Nlgn1), Shank2, and Shank3 mRNA levels and increased contactin-associated protein-like 2 mRNA level. The delay in neuronal maturation was also observed in Nlgn1-knockdown cells, which were transfected with Nlgn1 siRNA. These findings suggest that prenatal HDAC inhibition causes changes in gene expression of autism-related molecules linked to a delay of neuronal maturation. 相似文献
66.
Yukio Furukawa Yumi Inoue Aya Sakaguchi Yoko Mori Takuma Fukumura Tomoko Miyata Keiichi Namba Tohru Minamino 《Molecular microbiology》2016,102(3):405-416
FliS chaperone binds to flagellin FliC in the cytoplasm and transfers FliC to a sorting platform of the flagellar type III export apparatus through the interaction between FliS and FlhA for rapid and efficient protein export during flagellar filament assembly. FliS also suppresses the secretion of an anti‐σ factor, FlgM. Loss of FliS results in a short filament phenotype although the expression levels of FliC are increased considerably due to an increase in the secretion level of FlgM. Here to clarify the rate limiting step of FliC export in the absence of FliS, we isolated bypass mutants from a Salmonella ΔfliS mutant. All the bypass mutations were identified in FliC. These bypass mutations increased the export rate of FliC by ca. twofold, allowing the bypass mutant cells to produce longer filaments than the parental ΔfliS cells. Both far‐UV CD measurements and limited proteolysis revealed that the bypass mutations significantly destabilize the folded structure of FliC monomer. These results suggest that an unfolding step of FliC limits the export rate of FliC in the ΔfliS mutant, thereby producing short filaments. We propose that FliS promotes FliC docking at the FlhA platform to facilitate subsequent unfolding of FliC. 相似文献
67.
CRISPR/Cas9 is a novel tool for targeted mutagenesis and is applicable to plants, including rice. Previous reports on CRISPR/Cas9 in rice have demonstrated that target mutations are transmitted to the next generation in accordance with Mendelian law, but heritability of the target mutation and the role of inherited Cas9 gene have not been fully elucidated. Here, we targeted the rice phytoene desaturase gene, mutants of which exhibit an albino phenotype, by using CRISPR/Cas9 and analyzed segregation of target mutations. Agrobacterium-mediated methods using immature embryos successfully transformed a CRISPR/Cas9 system into five rice cultivars and subsequently induced mutation. Unpredicted segregations, with more mutants than theoretically predicted, were frequently found in T1 plants from monoallelic T0 mutants. Chimeric plants with both biallelic and monoallelic mutated cells were also observed in the T1. Next, we followed segregation of a target mutation in the T2 from monoallelic T1 mutants. When T1 mutants possessed Cas9, unpredicted segregations of the target mutation and chimeric plants were observed again in the T2. When T1 mutants did not possess Cas9, segregation of the target mutations followed Mendelian law and no chimeric plants appeared in the T2. T2 mutants with Cas9 had mutations different from the original ones found in T0. Our results indicated that inherited Cas9 was still active in later generations and could induce new mutations in the progeny, leading to chimerism and unpredicted segregation. We conclude that Cas9 has to be eliminated by segregation in T1 to generate homozygous mutants without chimerism or unpredicted segregation. 相似文献
68.
Toshimitsu Ishibashi T. Takizawa Hideaki Iwasaki Takuma Saito Shigeki Matsubara Eiko Nakazawa Kyotaro Kanazawa 《Histochemistry and cell biology》1999,112(3):221-232
We describe an improved copper ferrocyanide-based method for cytochemical detection of glucose-6-phosphate dehydrogenase (G6PD),
which was used to localize the enzyme within the ultrastructure of rat hepatocytes and adrenocortical cells. With this method,
glutaraldehyde fixation and the addition of exogenous electron carriers (for example, phenazine methosulfate) to the cytochemical
reaction medium were essential. Copper ferrocyanide reaction product showing the distribution of G6PD was readily recognized
at the light microscopic level as Hatchett’s brown staining and at the electron microscopic level as electron-dense deposits.
Within stained regions, enzyme cytochemical G6PD activity was found to be associated with ribosome-like structures. Because
G6PD is a soluble, cytosolic enzyme, its displacement or extraction may occur during conventional fixation. We, therefore,
combined a rapid-freezing technique with G6PD enzyme cytochemistry. The resultant rapid-freezing enzyme cytochemistry enabled
us to show the subcellular distribution of G6PD in a more life-like state; the localization of G6PD in rapidly frozen cells
was in substantial agreement with that in conventionally fixed cells.
Accepted: 14 July 1999 相似文献
69.
Noriaki Sasai Shogo Tada Jumi Ohshiro Chikara Kogiso Takuma Shinozuka 《Development, growth & differentiation》2024,66(1):89-100
During development, progenitor cell survival is essential for proper tissue functions, but the underlying mechanisms are not fully understood. Here we show that ERCC6L2, a member of the Snf2 family of helicase-like proteins, plays an essential role in the survival of developing chick neural cells. ERCC6L2 expression is induced by the Sonic Hedgehog (Shh) signaling molecule by a mechanism similar to that of the known Shh target genes Ptch1 and Gli1. ERCC6L2 blocks programmed cell death induced by Shh inhibition and this inhibition is independent of neural tube patterning. ERCC6L2 knockdown by siRNA resulted in the aberrant appearance of apoptotic cells. Furthermore, ERCC6L2 cooperates with the Shh signal and plays an essential role in the induction of the anti-apoptotic factor Bcl-2. Taken together, ERCC6L2 acts as a key factor in ensuring the survival of neural progenitor cells. 相似文献
70.
Masaki Honda Tatsuya Kudo Takahiro Kuwa Takuma Higashiura Tetsuya Fukaya Yoshinori Inoue 《Bioscience, biotechnology, and biochemistry》2017,81(2):365-371
Lycopene has a large number of geometric isomers caused by E/Z isomerization at arbitrary sites within the 11 conjugated double bonds, offering varying characteristics related to features such as antioxidant capacity and bioavailability. However, the geometric structures of only a few lycopene Z-isomers have been thoroughly identified from natural sources. In this study, seven multi-Z-isomers of lycopene, (9Z,13′Z)-, (5Z,13Z,9′Z)-, (9Z,9′Z)-, (5Z,13′Z)-, (5Z,9′Z)-, (5Z,9Z,5′Z)-, and (5Z,9Z)-lycopene, were obtained from tomato samples by thermal isomerization, and then isolated by elaborate chromatography, and fully assigned using proton nuclear magnetic resonance. Moreover, the theoretically preferred pathway from (all-E)-lycopene to di-Z-isomers was examined with a computational approach using a Gaussian program. Fine-tuning of the HPLC separation conditions led to the discovery of novel multi-Z-isomers, and whose formation was supported by advanced theoretical calculations. 相似文献