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Yasuyuki Aoyagi,Yasushi Saito,Masayuki Kuroda,Sakiyo Asada,Hideaki Bujo,Shigeaki Tanaka,Shunichi Konno,Masami Tanio,Itsuko Ishii,Masayuki Aso 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.3
The development of clinically applicable scaffolds is important for the application of cell transplantation in various human diseases. The aims of this study are to evaluate fibrin glue in a novel protein replacement therapy using proliferative adipocytes and to develop a mouse model system to monitor the delivery of the transgene product into the blood and the fate of the transduced cells after transplantation. Proliferative adipocytes from mouse adipose tissue were transduced by a retroviral vector harboring the human lecithin-cholesterol acyltransferase (lcat) gene, and were subcutaneously transplanted into mice combined with fibrin glue. The lcat gene transduction efficiency and the subsequent secretion of the product in mouse adipocytes were enhanced using a protamine concentration of 500 μg/ml. Adipogenesis induction did not significantly affect the lcat gene-transduced cell survival after transplantation. Immunohistochemistry showed the ectopic enzyme production to persist for 28 days in the subcutaneously transplanted genetransduced adipocytes. The increased viability of transplanted cells with fibrin glue was accompanied with the decrease in apoptotic cell death. The immunodetectable serum LCAT levels in mice implanted with the fibrin glue were comparable with those observed in mice implanted with Matrigel, indicating that the transplanted lcat gene-transduced adipocytes survived and functioned in the transplanted spaces with fibrin glue as well as with Matrigel for 28 days. Thus,this in vivo system using fibrin is expected to serve as a good model to further improve the transplanted cell/scaffold conditions for the stable and durable cell-based replacement of defective proteins in patients with LCAT deficiency.
Aoyagi, Yasuyuki,Kuroda, Masayuki,Asada, Sakiyo,Bujo, Hideaki,Tanaka, Shigeaki,Konno, Shunichi,Tanio, Masami,Ishii, Itsuko,Aso, Masayuki,Saito, Yasushi Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.3
The development of clinically applicable scaffolds is important for the application of cell transplantation in various human diseases. The aims of this study are to evaluate fibrin glue in a novel protein replacement therapy using proliferative adipocytes and to develop a mouse model system to monitor the delivery of the transgene product into the blood and the fate of the transduced cells after transplantation. Proliferative adipocytes from mouse adipose tissue were transduced by a retroviral vector harboring the human lecithin- cholesterol acyltransferase ($lcat$) gene, and were subcutaneously transplanted into mice combined with fibrin glue. The $lcat$ gene transduction efficiency and the subsequent secretion of the product in mouse adipocytes were enhanced using a protamine concentration of $500{\mu}g/ml$. Adipogenesis induction did not significantly affect the $lcat$ gene-transduced cell survival after transplantation. Immunohistochemistry showed the ectopic enzyme production to persist for 28 days in the subcutaneously transplanted genetransduced adipocytes. The increased viability of transplanted cells with fibrin glue was accompanied with the decrease in apoptotic cell death. The immunodetectable serum LCAT levels in mice implanted with the fibrin glue were comparable with those observed in mice implanted with Matrigel, indicating that the transplanted $lcat$ gene-transduced adipocytes survived and functioned in the transplanted spaces with fibrin glue as well as with Matrigel for 28 days. Thus, this $in$ $vivo$ system using fibrin is expected to serve as a good model to further improve the transplanted cell/scaffold conditions for the stable and durable cell-based replacement of defective proteins in patients with LCAT deficiency.
Primary Multiple Cardiac Myxomas in a Patient without the Carney Complex
Shohei Kataoka,Masato Otsuka,Masayuki Goto,Mitsuru Kahata,Asako Kumagai,Koji Inoue,Hiroshi Koganei,Kenji Enta,Yasuhiro Ishii 한국심초음파학회 2016 Journal of Cardiovascular Imaging (J Cardiovasc Im Vol.24 No.1
Cardiac tumors are rare, and multiple myxomas are even rarer. The latter phenomenon is mostly associated with the Carney complex, a dominantly inherited disease characterized by multiple primary cardiac myxomas, endocrinopathy, and spotty pigmentation of the skin. We report the rare case of a patient who did not have the Carney complex but had multiple primary cardiac tumors. A 78-year-old woman with a past history of breast cancer was referred to our hospital for further examination of multiple cardiac tumors. Echocardiography showed 4 tumors in the left atrium and left ventricle. We could not diagnose them preoperatively and decided to resect them surgically because they were mobile and could have caused embolism and obstruction. The postoperative pathological findings of all 4 tumors were myxomas, although the patient did not meet the diagnostic criteria of the Carney complex. Therefore, a rare case of multiple primary cardiac myxomas was diagnosed.
Kim, Raeyeong,Kanamaru, Shuji,Mikawa, Tsutomu,Pré,vost, Chantal,Ishii, Kentaro,Ito, Kentaro,Uchiyama, Susumu,Oda, Masayuki,Iwasaki, Hiroshi,Kim, Seog K,Takahashi, Masayuki Oxford University Press 2018 Nucleic acids research Vol.46 No.5
<P><B>Abstract</B></P><P>Mg<SUP>2+</SUP> ion stimulates the DNA strand exchange reaction catalyzed by RecA, a key step in homologous recombination. To elucidate the molecular mechanisms underlying the role of Mg<SUP>2+</SUP> and the strand exchange reaction itself, we investigated the interaction of RecA with Mg<SUP>2+</SUP> and sought to determine which step of the reaction is affected. Thermal stability, intrinsic fluorescence, and native mass spectrometric analyses of RecA revealed that RecA binds at least two Mg<SUP>2+</SUP> ions with K<SUB>D</SUB> ≈ 2 mM and 5 mM. Deletion of the C-terminal acidic tail of RecA made its thermal stability and fluorescence characteristics insensitive to Mg<SUP>2+</SUP> and similar to those of full-length RecA in the presence of saturating Mg<SUP>2+</SUP>. These observations, together with the results of a molecular dynamics simulation, support the idea that the acidic tail hampers the strand exchange reaction by interacting with other parts of RecA, and that binding of Mg<SUP>2+</SUP> to the tail prevents these interactions and releases RecA from inhibition. We observed that binding of the first Mg<SUP>2+</SUP> stimulated joint molecule formation, whereas binding of the second stimulated progression of the reaction. Thus, RecA is actively involved in the strand exchange step as well as bringing the two DNAs close to each other.</P>