A novel, simple, reliable, and sensitive method for multiple immunoenzyme staining: use of microwave oven heating to block antibody crossreactivity and retrieve antigensHY Lan, W Mu, DJ Nikolic-Paterson and RC Atkins Department of Nephrology, Monash Medical Centre, Clayton, Victoria, Australia. We report a simple and reliable method for detection of two or more antigens within tissue sections by indirect immunoenzyme staining using mouse monoclonal antibodies (MAbs). This technique involves treating sections with two 5-min microwave (MW) oven heatings between sequential rounds of three-layer immunoenzyme staining (mouse MAb, goat anti-mouse IgG, and mouse PAP or mouse APAAP) and color development. Discrete staining of cell surface, cytoplasmic, and nuclear antigens was evident within individual cells. This technique has a number of advantages over those currently available. First, MW treatment denatures bound antibody molecules, thereby completely blocking crossreactivity between sequential rounds of staining. This allows the use of primary (and other) antibodies raised in the same species and the use of a sensitive three-layer staining method. Second, antigen retrieval after MW treatment markedly increases the sensitivity of cytoplasmic and nuclear antigen detection. Third, inactivation of peroxidase and alkaline phosphatase enzymes present in PAP and APAAP complexes prevents inappropriate color development. Finally, this method can be used in both paraformaldehyde-fixed cryostat sections and formalin-fixed paraffin tissue sections. In conclusion, this is a simple, reliable, and sensitive technique that will be useful in many areas of diagnosis and research.
Volume 43,
Issue 1,
pp. 97-102,
01/01/1995
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R. R. Mundegar, E. Franke, R. Schafer, M. Zweyer, and A. Wernig Reduction of High Background Staining by Heating Unfixed Mouse Skeletal Muscle Tissue Sections Allows for Detection of Thermostable Antigens With Murine Monoclonal Antibodies J. Histochem. Cytochem., November 1, 2008; 56(11): 969 - 975. [Abstract] [Full Text] [PDF] |
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T. Arakawa, T. Masaki, T. Hirai, S. Doi, M. Kuratsune, K. Arihiro, N. Kohno, and N. Yorioka Activation of signal transducer and activator of transcription 3 correlates with cell proliferation and renal injury in human glomerulonephritis Nephrol. Dial. Transplant., November 1, 2008; 23(11): 3418 - 3426. [Abstract] [Full Text] [PDF] |
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J. S. Marino, B. J. Tausch, C. L. Dearth, M. V. Manacci, T. J. McLoughlin, S. J. Rakyta, M. P. Linsenmayer, and F. X. Pizza {beta}2-Integrins contribute to skeletal muscle hypertrophy in mice Am J Physiol Cell Physiol, October 1, 2008; 295(4): C1026 - C1036. [Abstract] [Full Text] [PDF] |
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W. Deng, S. W. Tsao, Y. K. Kwok, E. Wong, X. R. Huang, S. Liu, C. M. Tsang, H. Y.S. Ngan, A. N.Y. Cheung, H. Y. Lan, et al. Transforming Growth Factor {beta}1 Promotes Chromosomal Instability in Human Papillomavirus 16 E6E7-Infected Cervical Epithelial Cells Cancer Res., September 1, 2008; 68(17): 7200 - 7209. [Abstract] [Full Text] [PDF] |
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Y. Kobayashi, K. Suzuki, H. Kobayashi, S. Ohashi, K. Koike, P. Macchi, M. Kiebler, and K. Anzai C9orf10 Protein, a Novel Protein Component of Pur{alpha}-containing mRNA-protein Particles (Pur{alpha}-mRNPs): Characterization of Developmental and Regional Expressions in the Mouse Brain J. Histochem. Cytochem., August 1, 2008; 56(8): 723 - 731. [Abstract] [Full Text] [PDF] |
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J. W. Adams, J. Wang, J. R. Davis, C. Liaw, I. Gaidarov, J. Gatlin, N. D. Dalton, Y. Gu, J. Ross Jr., D. Behan, et al. Myocardial expression, signaling, and function of GPR22: a protective role for an orphan G protein-coupled receptor Am J Physiol Heart Circ Physiol, August 1, 2008; 295(2): H509 - H521. [Abstract] [Full Text] [PDF] |
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X. R. Huang, A. C. K. Chung, X. J. Wang, K. N. Lai, and H. Y. Lan Mice overexpressing latent TGF-{beta}1 are protected against renal fibrosis in obstructive kidney disease Am J Physiol Renal Physiol, July 1, 2008; 295(1): F118 - F127. [Abstract] [Full Text] [PDF] |
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C. M. van der Loos Multiple Immunoenzyme Staining: Methods and Visualizations for the Observation With Spectral Imaging J. Histochem. Cytochem., April 1, 2008; 56(4): 313 - 328. [Abstract] [Full Text] [PDF] |
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X. R. Huang, A. C.K. Chung, L. Zhou, X. J. Wang, and H. Y. Lan Latent TGF-{beta}1 Protects Against Crescentic Glomerulonephritis J. Am. Soc. Nephrol., February 1, 2008; 19(2): 233 - 242. [Abstract] [Full Text] [PDF] |
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Z. E. Toth and E. Mezey Simultaneous Visualization of Multiple Antigens With Tyramide Signal Amplification Using Antibodies From the Same Species J. Histochem. Cytochem., June 1, 2007; 55(6): 545 - 554. [Abstract] [Full Text] [PDF] |
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P. Fu, F. Liu, S. Su, W. Wang, X. R. Huang, M. L. Entman, R. J. Schwartz, L. Wei, and H. Y. Lan Signaling Mechanism of Renal Fibrosis in Unilateral Ureteral Obstructive Kidney Disease in ROCK1 Knockout Mice J. Am. Soc. Nephrol., November 1, 2006; 17(11): 3105 - 3114. [Abstract] [Full Text] [PDF] |
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K N Naresh, I Lampert, R Hasserjian, D Lykidis, K Elderfield, D Horncastle, N Smith, W Murray-Brown, and G W Stamp Optimal processing of bone marrow trephine biopsy: the Hammersmith Protocol. J. Clin. Pathol., September 1, 2006; 59(9): 903 - 911. [Abstract] [Full Text] [PDF] |
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V. Koka, W. Wang, X. R. Huang, S. Kim-Mitsuyama, L. D. Truong, and H. Y. Lan Advanced Glycation End Products Activate a Chymase-Dependent Angiotensin II-Generating Pathway in Diabetic Complications Circulation, March 14, 2006; 113(10): 1353 - 1360. [Abstract] [Full Text] [PDF] |
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E. Sato, S. H. Olson, J. Ahn, B. Bundy, H. Nishikawa, F. Qian, A. A. Jungbluth, D. Frosina, S. Gnjatic, C. Ambrosone, et al. Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer PNAS, December 20, 2005; 102(51): 18538 - 18543. [Abstract] [Full Text] [PDF] |
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J. B. de Haan, N. Stefanovic, D. Nikolic-Paterson, L. L. Scurr, K. D. Croft, T. A. Mori, P. Hertzog, I. Kola, R. C. Atkins, and G. H. Tesch Kidney expression of glutathione peroxidase-1 is not protective against streptozotocin-induced diabetic nephropathy Am J Physiol Renal Physiol, September 1, 2005; 289(3): F544 - F551. [Abstract] [Full Text] [PDF] |
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W. Wang, X. R. Huang, A. G. Li, F. Liu, J.-H. Li, L. D. Truong, X. J. Wang, and H. Y. Lan Signaling Mechanism of TGF-{beta}1 in Prevention of Renal Inflammation: Role of Smad7 J. Am. Soc. Nephrol., May 1, 2005; 16(5): 1371 - 1383. [Abstract] [Full Text] [PDF] |
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Y.-Z. Kong, X.-R. Huang, X. Ouyang, J.-J. Tan, G. Fingerle-Rowson, M. Bacher, W. Mu, L. A. Scher, L. Leng, R. Bucala, et al. Evidence for vascular macrophage migration inhibitory factor in destabilization of human atherosclerotic plaques Cardiovasc Res, January 1, 2005; 65(1): 272 - 282. [Abstract] [Full Text] [PDF] |
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A. Navarro, E. del Valle, and J. Tolivia Differential Expression of Apolipoprotein D in Human Astroglial and Oligodendroglial Cells J. Histochem. Cytochem., August 1, 2004; 52(8): 1031 - 1036. [Abstract] [Full Text] [PDF] |
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Y. Ikezumi, L. Hurst, R. C. Atkins, and D. J. Nikolic-Paterson Macrophage-Mediated Renal Injury Is Dependent on Signaling via the JNK Pathway J. Am. Soc. Nephrol., July 1, 2004; 15(7): 1775 - 1784. [Abstract] [Full Text] [PDF] |
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T. Masaki, C. Stambe, P. A. Hill, J. Dowling, R. C. Atkins, and D. J. Nikolic-Paterson Activation of the Extracellular-Signal Regulated Protein Kinase Pathway in Human Glomerulopathies J. Am. Soc. Nephrol., July 1, 2004; 15(7): 1835 - 1843. [Abstract] [Full Text] [PDF] |
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J. H. Li, W. Wang, X. R. Huang, M. Oldfield, A. M. Schmidt, M. E. Cooper, and H. Y. Lan Advanced Glycation End Products Induce Tubular Epithelial-Myofibroblast Transition through the RAGE-ERK1/2 MAP Kinase Signaling Pathway Am. J. Pathol., April 1, 2004; 164(4): 1389 - 1397. [Abstract] [Full Text] [PDF] |
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S. La Rosa, S. Uccella, S. Marchet, C. Capella, and R. V. Lloyd Localization of Inhibins and Activins in Normal Endocrine Cells and Endocrine Tumors of the Gut and Pancreas: an Immunohistochemical and In Situ Hybridization Study J. Histochem. Cytochem., February 1, 2004; 52(2): 217 - 225. [Abstract] [Full Text] [PDF] |
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C. Stambe, D. J. Nikolic-Paterson, P. A. Hill, J. Dowling, and R. C. Atkins p38 Mitogen-Activated Protein Kinase Activation and Cell Localization in Human Glomerulonephritis: Correlation with Renal Injury J. Am. Soc. Nephrol., February 1, 2004; 15(2): 326 - 336. [Abstract] [Full Text] [PDF] |
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C. Stambe, R. C. Atkins, G. H. Tesch, T. Masaki, G. F. Schreiner, and D. J. Nikolic-Paterson The Role of p38{alpha} Mitogen-Activated Protein Kinase Activation in Renal Fibrosis J. Am. Soc. Nephrol., February 1, 2004; 15(2): 370 - 379. [Abstract] [Full Text] [PDF] |
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E. del Valle, A. Navarro, A. Astudillo, and J. Tolivia Apolipoprotein D Expression in Human Brain Reactive Astrocytes J. Histochem. Cytochem., October 1, 2003; 51(10): 1285 - 1290. [Abstract] [Full Text] [PDF] |
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K. Hasui, T. Takatsuka, R. Sakamoto, S. Matsushita, S.-i. Tsuyama, S. Izumo, and F. Murata Double Autoimmunostaining with Glycine Treatment J. Histochem. Cytochem., September 1, 2003; 51(9): 1169 - 1176. [Abstract] [Full Text] [PDF] |
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T. Uchihara, A. Nakamura, H. Nakayama, K. Arima, N. Ishizuka, H. Mori, and S. Mizushima Triple Immunofluorolabeling with Two Rabbit Polyclonal Antibodies and a Mouse Monoclonal Antibody Allowing Three-dimensional Analysis of Cotton Wool Plaques in Alzheimer Disease J. Histochem. Cytochem., September 1, 2003; 51(9): 1201 - 1206. [Abstract] [Full Text] [PDF] |
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Z.-H. Liu, S.-F. Chen, H. Zhou, H.-P. Chen, and L.-S. Li Glomerular expression of C-C chemokines in different types of human crescentic glomerulonephritis Nephrol. Dial. Transplant., August 1, 2003; 18(8): 1526 - 1534. [Abstract] [Full Text] [PDF] |
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Z.-H. Liu, S.-F. Chen, H. Zhou, H.-P. Chen, and L.-S. Li Glomerular expression of C-C chemokines in different types of human crescentic glomerulonephritis Nephrol. Dial. Transplant., August 1, 2003; 18(88): 1526 - 1534. [Abstract] [Full Text] |
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X. R. Huang, W. Y. Chen, L. D. Truong, and H. Y. Lan Chymase Is Upregulated in Diabetic Nephropathy: Implications for an Alternative Pathway of Angiotensin II-Mediated Diabetic Renal and Vascular Disease J. Am. Soc. Nephrol., July 1, 2003; 14(7): 1738 - 1747. [Abstract] [Full Text] [PDF] |
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H. Y. Lan, W. Mu, N. Tomita, X. R. Huang, J. H. Li, H.-J. Zhu, R. Morishita, and R. J. Johnson Inhibition of Renal Fibrosis by Gene Transfer of Inducible Smad7 Using Ultrasound-Microbubble System in Rat UUO Model J. Am. Soc. Nephrol., June 1, 2003; 14(6): 1535 - 1548. [Abstract] [Full Text] [PDF] |
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C.-M. Yu, K. W.-H. Lai, Y.-X. Chen, X.-R. Huang, and H. Y. Lan Expression of Macrophage Migration Inhibitory Factor in Acute Ischemic Myocardial Injury J. Histochem. Cytochem., May 1, 2003; 51(5): 625 - 631. [Abstract] [Full Text] [PDF] |
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Y. Ikezumi, R. C. Atkins, and D. J. Nikolic-Paterson Interferon-{gamma} Augments Acute Macrophage-Mediated Renal Injury Via a Glucocorticoid-Sensitive Mechanism J. Am. Soc. Nephrol., April 1, 2003; 14(4): 888 - 898. [Abstract] [Full Text] [PDF] |
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H. Sipahutar, P. Sourdaine, S. Moslemi, B. Plainfosse, and G.-E. Seralini Immunolocalization of Aromatase in Stallion Leydig Cells and Seminiferous Tubules J. Histochem. Cytochem., March 1, 2003; 51(3): 311 - 318. [Abstract] [Full Text] [PDF] |
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J. M. Ricono, Y.-C. Xu, M. Arar, D.-c. Jin, J. L. Barnes, and H. E. Abboud Morphological Insights into the Origin of Glomerular Endothelial and Mesangial Cells and Their Precursors J. Histochem. Cytochem., February 1, 2003; 51(2): 141 - 150. [Abstract] [Full Text] [PDF] |
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C. Stambe, R. C. Atkins, G. H. Tesch, A. M. Kapoun, P. A. Hill, G. F. Schreiner, and D. J. Nikolic-Paterson Blockade of p38{alpha} MAPK Ameliorates Acute Inflammatory Renal Injury in Rat Anti-GBM Glomerulonephritis J. Am. Soc. Nephrol., February 1, 2003; 14(2): 338 - 351. [Abstract] [Full Text] [PDF] |
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E. C. Ibrahim, Y. Allory, F. Commo, B. Gattegno, P. Callard, and P. Paul Altered Pattern of Major Histocompatibility Complex Expression in Renal Carcinoma: Tumor-Specific Expression of the Nonclassical Human Leukocyte Antigen-G Molecule Is Restricted to Clear Cell Carcinoma While Up-Regulation of Other Major Histocompatibility Complex Antigens Is Primarily Distributed in All Subtypes of Renal Carcinoma Am. J. Pathol., February 1, 2003; 162(2): 501 - 508. [Abstract] [Full Text] [PDF] |
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S.-L. Lin, Y.-M. Chen, C.-T. Chien, W.-C. Chiang, C.-C. Tsai, and T.-J. Tsai Pentoxifylline Attenuated the Renal Disease Progression in Rats with Remnant Kidney J. Am. Soc. Nephrol., December 1, 2002; 13(12): 2916 - 2929. [Abstract] [Full Text] [PDF] |
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C. G. Ventura, T. M. Coimbra, S. B. de Campos, I. de Castro, L. Yu, and A. C. Seguro Mycophenolate Mofetil Attenuates Renal Ischemia/Reperfusion Injury J. Am. Soc. Nephrol., October 1, 2002; 13(10): 2524 - 2533. [Abstract] [Full Text] [PDF] |
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Y. Le Meur, G. H. Tesch, P. A. Hill, W. Mu, R. Foti, D. J. Nikolic-Paterson, and R. C. Atkins Macrophage accumulation at a site of renal inflammation is dependent on the M-CSF/c-fms pathway J. Leukoc. Biol., September 1, 2002; 72(3): 530 - 537. [Abstract] [Full Text] [PDF] |
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J. G. Ramos, J. Varayoud, V. L. Bosquiazzo, E. H. Luque, and M. Munoz-de-Toro Cellular Turnover in the Rat Uterine Cervix and Its Relationship to Estrogen and Progesterone Receptor Dynamics Biol Reprod, September 1, 2002; 67(3): 735 - 742. [Abstract] [Full Text] [PDF] |
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D. Eladari, L. Cheval, F. Quentin, O. Bertrand, I. Mouro, B. Cherif-Zahar, J.-P. Cartron, M. Paillard, A. Doucet, and R. Chambrey Expression of RhCG, a New Putative NH3/NH4+ Transporter, along the Rat Nephron J. Am. Soc. Nephrol., August 1, 2002; 13(8): 1999 - 2008. [Abstract] [Full Text] [PDF] |
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F. G. Brown, D. J. Nikolic-Paterson, P. A. Hill, N. M. Isbel, J. Dowling, C. M. Metz, and R. C. Atkins Urine Macrophage Migration Inhibitory Factor Reflects the Severity of Renal Injury in Human Glomerulonephritis J. Am. Soc. Nephrol., January 1, 2002; 13(90001): S7 - 13. [Abstract] [Full Text] [PDF] |
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K. N. LAI, F. K. LI, H. YUI LAN, S. TANG, A. W. L. TSANG, D. T. M. CHAN, and J. C. LEUNG Expression of Aquaporin-1 in Human Peritoneal Mesothelial Cells and Its Upregulation by Glucose In Vitro J. Am. Soc. Nephrol., May 1, 2001; 12(5): 1036 - 1045. [Abstract] [Full Text] |
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R. Tordjman, S. Delaire, J. Plouet, S. Ting, P. Gaulard, S. Fichelson, P.-H. Romeo, and V. Lemarchandel Erythroblasts are a source of angiogenic factors Blood, April 1, 2001; 97(7): 1968 - 1974. [Abstract] [Full Text] [PDF] |
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S.-G. Lin, X.-Y. Yu, Y.-X. Chen, X. R. Huang, C. Metz, R. Bucala, C.-P. Lau, and H. Y. Lan De Novo Expression of Macrophage Migration Inhibitory Factor in Atherogenesis in Rabbits Circ. Res., December 8, 2000; 87(12): 1202 - 1208. [Abstract] [Full Text] [PDF] |
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K. Chen, Y. Wei, G. C. Sharp, and H. Braley-Mullen Characterization of thyroid fibrosis in a murine model of granulomatous experimental autoimmune thyroiditis J. Leukoc. Biol., December 1, 2000; 68(6): 828 - 835. [Abstract] [Full Text] |
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A. SHIMIZU, K. YAMADA, S. M. MEEHAN, D. H. SACHS, and R. B. COLVIN Acceptance Reaction: Intragraft Events Associated with Tolerance to Renal Allografts in Miniature Swine J. Am. Soc. Nephrol., December 1, 2000; 11(12): 2371 - 2380. [Abstract] [Full Text] |
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M. P. RASTALDI, F. FERRARIO, A. CRIPPA, G. DELL'ANTONIO, D. CASARTELLI, C. GRILLO, and G. D'AMICO Glomerular Monocyte-Macrophage Features in ANCA-Positive Renal Vasculitis and Cryoglobulinemic Nephritis J. Am. Soc. Nephrol., November 1, 2000; 11(11): 2036 - 2043. [Abstract] [Full Text] |
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X. R. HUANG, A. R. KITCHING, P. G. TIPPING, and S. R. HOLDSWORTH Interleukin-10 Inhibits Macrophage-Induced Glomerular Injury J. Am. Soc. Nephrol., February 1, 2000; 11(2): 262 - 269. [Abstract] [Full Text] [PDF] |
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F.-J. Lai, S.-J. Shin, Y.-J. Lee, S.-R. Lin, W.-Y. Jou, and J.-H. Tsai Up-Regulation of Adrenal Cortical and Medullary Atrial Natriuretic Peptide and Gene Expression in Rats with Deoxycorticosterone Acetate-Salt Treatment Endocrinology, January 1, 2000; 141(1): 325 - 332. [Abstract] [Full Text] [PDF] |
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Y.-Y. Ng, J.-M. Fan, W. Mu, D. J. Nikolic-Paterson, W.-C. Yang, T.-p. Huang, R. C. Atkins, and H. Y. Lan Glomerular epithelial-myofibroblast transdifferentiation in the evolution of glomerular crescent formation Nephrol. Dial. Transplant., December 1, 1999; 14(12): 2860 - 2872. [Abstract] [Full Text] [PDF] |
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X. Q. Yu, J.-M. Fan, D. J. Nikolic-Paterson, N. Yang, W. Mu, R. Pichler, R. J. Johnson, R. C. Atkins, and H. Y. Lan IL-1 Up-Regulates Osteopontin Expression in Experimental Crescentic Glomerulonephritis in the Rat Am. J. Pathol., March 1, 1999; 154(3): 833 - 841. [Abstract] [Full Text] [PDF] |
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Y. Takahashi, C. Murai, S. Shibata, Y. Munakata, T. Ishii, K. Ishii, T. Saitoh, T. Sawai, K. Sugamura, and T. Sasaki Human parvovirus B19 as a causative agent for rheumatoid arthritis PNAS, July 7, 1998; 95(14): 8227 - 8232. [Abstract] [Full Text] [PDF] |
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B. G. Short, D. M. Zimmerman, and L. W. Schwartz Automated Double Labeling of Proliferation and Apoptosis in Glutathione S-transferase-positive Hepatocytes in Rats J. Histochem. Cytochem., September 1, 1997; 45(9): 1299 - 1306. [Abstract] [Full Text] [PDF] |
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H. Y. Lan, M. Bacher, N. Yang, W. Mu, D. J. Nikolic-Paterson, C. Metz, A. Meinhardt, R. Bucala, and R. C. Atkins The Pathogenic Role of Macrophage Migration Inhibitory Factor in Immunologically Induced Kidney Disease in the Rat J. Exp. Med., April 21, 1997; 185(8): 1455 - 1466. [Abstract] [Full Text] [PDF] |
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S.-R. Shi, R. J. Cote, and C. R. Taylor Antigen Retrieval Immunohistochemistry: Past, Present, and Future J. Histochem. Cytochem., March 1, 1997; 45(3): 327 - 344. [Abstract] [Full Text] [PDF] |
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