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991.
Mass spectrometric identification of citrullination sites and immunohistochemical detection of citrullinated glial fibrillary acidic protein in Alzheimer's disease brains 下载免费PDF全文
Akihito Ishigami Hirofumi Masutomi Setsuko Handa Megumi Nakamura Shuuichi Nakaya Yoshiaki Uchida Yuko Saito Shigeo Murayama Byungki Jang Yong‐Chul Jeon Eun‐Kyoung Choi Yong‐Sun Kim Yasushi Kasahara Naoki Maruyama Tosifusa Toda 《Journal of neuroscience research》2015,93(11):1664-1674
Peptidylarginine deiminases (PADs) are posttranslational modification enzymes that convert protein arginine to citrulline residues in a calcium ion‐dependent manner. Previously, we reported the abnormal accumulation of citrullinated proteins and the increase in the amount of PAD2 in hippocampi from Alzheimer's disease (AD) patients. Moreover, glial fibrillary acidic protein (GFAP), an astrocyte‐specific marker protein, and vimentin were identified as citrullinated proteins by using two‐dimensional gel electrophoresis and MALDI‐TOF mass spectrometry. To clarify the substrate specificity of PADs against GFAP, we prepared recombinant human (rh)PAD1, rhPAD2, rhPAD3, rhPAD4, and rhGFAP. After incubation of rhGFAP with rhPAD1, rhPAD2, rhPAD3, and rhPAD4, citrullinated (cit‐)rhGFAP was detected by Western blotting. The citrullination of rhGFAP by rhPAD2 was unique, specific, and time dependent; additionally, rhPAD1 slightly citrullinated rhGFAP. We then generated eight anti‐cit‐rhGFAP monoclonal antibodies, CTGF‐125, ?128, ?129, ?1212, ?1213, ?1221, ?122R, and ?1224R, which reacted specifically with cit‐rhGFAP. Two of those eight monoclonal antibodies, CTGF‐122R and ?1224R, reacted with both cit‐rhGFAP and rhGFAP in Western blots. By using the CTGF‐1221 antibody and a tandem mass spectrometer, we identified the two independent citrullination sites (R270Cit and R416Cit) of cit‐rhGFAP. Immunohistochemical analysis with CTGF‐1221 antibody revealed cit‐GFAP staining in the hippocampus of AD brain, and the cit‐GFAP‐positive cells appeared to be astrocyte‐like cells. These collective results strongly suggest that PAD2 is responsible for the citrullination of GFAP in the progression of AD and that the monoclonal antibody CTGF‐1221, reacting with cit‐GFAP at R270Cit and R416Cit, is useful for immunohistochemical investigation of AD brains. © 2015 Wiley Periodicals, Inc. 相似文献
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Yoshiyuki Tsukamoto Shoichi Fumoto Tsuyoshi Noguchi Kazuyoshi Yanagihara Yuka Hirashita Chisato Nakada Naoki Hijiya Tomohisa Uchida Keiko Matsuura Ryoji Hamanaka Kazunari Murakami Masao Seto Masafumi Inomata Masatsugu Moriyama 《American journal of cancer research》2015,5(10):2998-3014
Previously, we have reported that gain at chromosome 20q13 is the most common genomic copy number aberration in gastric cancer (GC) (29/30 cases), and that among the genes located in this region, we have identified DDX27, whose expression level shows the highest correlation with genomic copy number, as a candidate therapeutic target for GC. Here, we analyzed the clinicopathological significance of DDX27 using immunohistochemistry and studied its functions using knockdown assays. We found that DDX27 was frequently upregulated in GC tissues (98 of 140 cases, 70%), and significantly associated with venous invasion and liver metastasis. Furthermore, multivariate analysis of GC patients showed that high expression of DDX27 was independently associated with poorer prognosis. In functional assays, knockdown of DDX27 reduced the ability of GC cells to form colonies both on conventional plates and soft agar, but had little effect on their invasiveness. We also found that knockdown of DDX27 reduced the viability of GC cells through inhibition of cell cycle progression independently of apoptosis. Interestingly, DDX27 depletion induced accumulation of TP53 in a TP53 wild-type cell line, AGS, but not in a TP53-deleted cell line, 44As3, although DDX27 knockdown commonly reduced the viability of both, indicating the TP53-dependent and independent cell cycle control of DDX27. Thus, our results suggest that expression of DDX27 contributes to colony formation by GC cells through cell cycle control and may be a potential therapeutic target for GC patients with chromosome gain at 20q13. 相似文献
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Sakurazawa N Kato S Fujita I Kanazawa Y Onodera H Uchida E 《World journal of gastrointestinal endoscopy》2012,4(6):231-235
The indications for endoscopic treatment have expanded in recent years,and relatively intestinal-type mucosal stomach carcinomas with a low potential for metastasis are now often resected en bloc by endoscopic submucosal dissection(ESD),even if they measure over 20 mm in size.However,ESD requires complex maneuvers,which entails a long operation time,and is often accompanied by complications such as bleeding and perforation.Many technical developments have been implemented to overcome these complications.The scope,cutting device,hemostasis device,and other supportive devices have been improved.However,even with these innovations,ESD remains a potentially complex procedure.One of the major difficulties is poor visualization of the submucosal layer resulting from the poor countertraction afforded during submucosal dissection.Recently,countertraction devices have been developed.In this paper,we introduce countertraction techniques and devices mainly for gastric cancer. 相似文献
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Ohara H Okazaki K Tsubouchi H Inui K Kawa S Kamisawa T Tazuma S Uchida K Hirano K Yoshida H Nishino T Ko SB Mizuno N Hamano H Kanno A Notohara K Hasebe O Nakazawa T Nakanuma Y Takikawa H 《Journal of hepato-biliary-pancreatic sciences》2012,19(5):536-542
Background
IgG4-sclerosing cholangitis (IgG4-SC) patients have an increased level of serum IgG4, dense infiltration of IgG4-positive plasma cells with extensive fibrosis in the bile duct wall, and a good response to steroid therapy. However, it is not easy to distinguish IgG4-SC from primary sclerosing cholangitis, pancreatic cancer, and cholangiocarcinoma on the basis of cholangiographic findings alone because various cholangiographic features of IgG4-SC are similar to those of the above progressive or malignant diseases.Methods
The Research Committee of IgG4-related Diseases and the Research Committee of Intractable Diseases of Liver and Biliary Tract in association with the Ministry of Health, Labor and Welfare, Japan and the Japan Biliary Association have set up a working group consisting of researchers specializing in IgG4-SC, and established the new clinical diagnostic criteria of IgG4-SC 2012.Results
The diagnosis of IgG4-SC is based on the combination of the following 4 criteria: (1) characteristic biliary imaging findings, (2) elevation of serum IgG4 concentrations, (3) the coexistence of IgG4-related diseases except those of the biliary tract, and (4) characteristic histopathological features. Furthermore, the effectiveness of steroid therapy is an optional extra diagnostic criterion to confirm accurate diagnosis of IgG4-SC.Conclusion
These diagnostic criteria for IgG4-SC are useful in practice for general physicians and other nonspecialists. 相似文献999.
Atsuro Uchida Kei Shinoda Celso Soiti Matsumoto Miho Kawai Sayaka Kawai Hisao Ohde Yoko Ozawa Susumu Ishida Makoto Inoue Atsushi Mizota Kazuo Tsubota 《Case reports in ophthalmology》2012,3(3):396-405
Purpose
To present our findings on the cause of an acute visual field defect (VFD) that developed in a patient on the day after vitrectomy for proliferative diabetic retinopathy.Case
A 50-year-old man complained of a blind area in the superior visual field that developed one day after vitrectomy. The patient had undergone uncomplicated vitrectomy for a long-duration vitreous hemorrhage associated with proliferative diabetic retinopathy. Residual vitreous hemorrhage hampered a clear view of the fundus. Goldmann perimetry showed a horizontal VFD in the superior field. The area corresponding to the VFD was examined by multifocal electroretinograms (mfERGs) and multifocal visual evoked potentials (mfVEPs). The amplitudes of the mfVEPs were reduced with prolonged implicit times especially when the superior hemifield was stimulated, while the amplitudes and implicit times were within the normal range when other parts of the visual field were stimulated. In addition, the full-field photopic ERGs and photopic negative responses were attenuated in the right eye. These findings suggested that the VFD did not originate from alterations in the retinal inner and middle layer but in the ganglion cells. The visual acuity improved to 1.2 but his optic disc became pale and the VFD remained unchanged more than 12 years after the surgery.Conclusion
We suggest that vitrectomy can cause ischemic optic neuropathy by interfering with the circulation associated with diabetes mellitus. Evaluations by mfERGs, mfVEPs, and full-field photopic ERGs were helpful in making the diagnosis.Key words: Ischemic optic neuropathy, Proliferative diabetic retinopathy, Multifocal electroretinogram, Multifocal visual evoked potentials, Photopic negative response 相似文献1000.