23 results on '"Kristen J. Kelynack"'
Search Results
2. An In Vitro Murine Model of Vascular Smooth Muscle Cell Mineralization
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Kristen J, Kelynack and Stephen G, Holt
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Calcium Phosphates ,Staining and Labeling ,Cell Culture Techniques ,In Vitro Techniques ,Muscle, Smooth, Vascular ,Cell Line ,Phosphates ,Disease Models, Animal ,Mice ,Calcification, Physiologic ,Animals ,Calcium ,Colorimetry ,Vascular Calcification - Abstract
Vascular calcification (VC) is seen ubiquitously in aging blood vessels and prematurely in disease states like renal failure. It is thought to be driven by a number of systemic and local factors that lead to extra-osseous deposition of mineral in the vascular wall and valves as a common endpoint. The response of resident vascular smooth muscle cell to these dystrophic signals appears to be important in this process. Whilst in vivo models allow the observation of global changes in a pro-calcific environment, identifying the specific cells and mechanisms involved has been largely garnered from in vitro experiments, which provide added benefits in terms of reproducibility, cost, and convenience. Here we describe a 7-21 day cell culture model of calcification developed using immortalized murine vascular smooth muscle cells (MOVAS-1). This model provides a method by which vascular smooth muscle cell involvement and manipulation within a mineralizing domain can be studied.
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- 2015
3. An In Vitro Murine Model of Vascular Smooth Muscle Cell Mineralization
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Kristen J. Kelynack and Stephen G Holt
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0301 basic medicine ,Vascular smooth muscle ,Cell ,chemistry.chemical_element ,Anatomy ,Calcium ,Biology ,medicine.disease ,Mineralization (biology) ,In vitro ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,Cell culture ,In vivo ,medicine ,Calcification - Abstract
Vascular calcification (VC) is seen ubiquitously in aging blood vessels and prematurely in disease states like renal failure. It is thought to be driven by a number of systemic and local factors that lead to extra-osseous deposition of mineral in the vascular wall and valves as a common endpoint. The response of resident vascular smooth muscle cell to these dystrophic signals appears to be important in this process. Whilst in vivo models allow the observation of global changes in a pro-calcific environment, identifying the specific cells and mechanisms involved has been largely garnered from in vitro experiments, which provide added benefits in terms of reproducibility, cost, and convenience. Here we describe a 7-21 day cell culture model of calcification developed using immortalized murine vascular smooth muscle cells (MOVAS-1). This model provides a method by which vascular smooth muscle cell involvement and manipulation within a mineralizing domain can be studied.
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- 2015
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4. Effect of Inhibition of Farnesylation and Geranylgeranylation on Renal Fibrogenesis in vitro
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Tim D. Hewitson, Kristen J. Kelynack, Rosemary Masterson, and Gavin J. Becker
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Nephrology ,medicine.medical_specialty ,Time Factors ,Physiology ,Protein Prenylation ,Apoptosis ,Rho family of GTPases ,In Vitro Techniques ,Kidney ,Collagen Type I ,Immediate-Early Proteins ,Geranylgeranylation ,Prenylation ,Internal medicine ,Genetics ,medicine ,Animals ,Farnesyltranstransferase ,RNA, Messenger ,Enzyme Inhibitors ,Fibroblast ,Cytoskeleton ,Cell Proliferation ,Alkyl and Aryl Transferases ,biology ,Farnesyltransferase inhibitor ,Connective Tissue Growth Factor ,General Medicine ,Fibroblasts ,Fibrosis ,In vitro ,Rats ,Cell biology ,Collagen Type I, alpha 1 Chain ,Actin Cytoskeleton ,Kinetics ,medicine.anatomical_structure ,Biochemistry ,Benzamides ,ras Proteins ,biology.protein ,Intercellular Signaling Peptides and Proteins ,Collagen ,Guanosine Triphosphate - Abstract
Background: The Ras and Rho family of GTPases serve as essential molecular switches in the downstream signalling of many cytokines involved in the regulation of renal fibroblast activity. Prenylation is a post-translational process critical to the membrane localization and function of these GTPases. We studied the effects of a farnesyltransferase inhibitor BMS-191563 and geranylgeranyltransferase inhibitor GGTI-298 on renal fibrogenesis in vitro. Methods: Functional studies examined the effects of BMS-191563 and GGTI-298 on rat renal fibroblast kinetics, collagen synthesis and collagen gel contraction. Pro-collagen α1(I) mRNA expression was measured by Northern analysis and CTGF expression by Western blotting. Results: Fibroblast proliferation was significantly reduced by both agents. Exposure of fibroblasts to BMS-191563 resulted in a significant reduction in total collagen production and pro-collagen α1(I) mRNA expression, an effect also observed but to a lesser degree with GGTI-298. Both agents significantly reduced CTGF protein expression. Fibroblast-mediated collagen I lattice contraction was decreased at 48 h by GGTI-298, an effect not observed with BMS-191563. Consistent with this finding, marked actin filament disassembly was evident by phalloidin staining of fibroblasts exposed to GGTI-298. Conclusion: BMS-191563 and GGTI-298 exhibit different effects on renal fibroblast function reflecting their predominant roles in inhibiting prenylation of Ras or Rho proteins respectively. Further studies are warranted to establish their potential therapeutic application in the treatment of progressive renal disease.
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- 2005
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5. Intracellular Cyclic Nucleotide Analogues Inhibit in vitro Mitogenesis and Activation of Fibroblasts Derived from Obstructed Rat Kidneys
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Kristen J. Kelynack, Gavin J. Becker, Marina Martic, Melanie G. Tait, Teresa Bisucci, Ian A. Darby, and Tim D. Hewitson
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medicine.medical_specialty ,Physiology ,8-Bromo Cyclic Adenosine Monophosphate ,Biology ,Kidney ,Cell morphology ,Immediate-Early Proteins ,Cyclic nucleotide ,chemistry.chemical_compound ,Internal medicine ,Cyclic AMP ,Genetics ,medicine ,Animals ,Nucleotide ,Fibroblast ,Cyclic GMP ,Cells, Cultured ,chemistry.chemical_classification ,Connective Tissue Growth Factor ,DNA ,General Medicine ,Fibroblasts ,Fibrosis ,Actins ,Rats ,CTGF ,Blot ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Nephrology ,Intercellular Signaling Peptides and Proteins ,Collagen ,Nucleotides, Cyclic ,Myofibroblast ,Cell Division ,Intracellular ,Ureteral Obstruction - Abstract
As several studies indirectly suggest that inhibiting the intracellular breakdown of cyclic nucleotides may inhibit fibrogenesis, this study used membrane permeable cyclic nucleotide analogues to examine the role of cAMP and cGMP signaling pathways in the regulation of renal fibroblast function. Fibroblasts were isolated by explant outgrowth culture of rat kidneys post unilateral ureteric obstruction. Subcultured cells were exposed to 10– 1,000 µM of the cyclic nucleotide analogues 8-bromo-cAMP (8br-cAMP) and 8-bromo-cGMP (8br-cGMP). Functional parameters examined included mitogenesis (thymidine incorporation), collagen synthesis (proline incorporation), myofibroblast differentiation (Western blotting for α-smooth muscle actin; α-SMA) and expression of CTGF (Northern blotting), a TGF-β1-driven immediate early response gene. Serum-stimulated mitogenesis was decreased 27 ± 4% by 100 µM 8br-cAMP (p < 0.01), 49 ± 6% by 1,000 µM 8br-cAMP (p < 0.001) and 43 ± 7% by 1,000 µM 8br-cGMP (p < 0.01). 1,000 µM 8br-cAMP and 8br-cGMP reduced basal collagen synthesis by 80 ± 5 and 60 ± 21% respectively (both p < 0.05). Maximum dose of 8br-cAMP but not 8br-cGMP inhibited basal expression of the differentiation marker α-SMA by 43 ± 33 (p < 0.05), resulted in a more rounded cell morphology and reduced expression of CTGF by 39 ± 24% (p < 0.05). Measurement of mitochondrial activity confirmed that effects were independent of cell toxicity. In conclusion, cyclic nucleotides inhibit fibrogenesis in vitro. Strategies which elevate intracellular cyclic nucleotide concentrations may therefore be therapeutically valuable in preventing the proliferation and activation of fibroblasts in progressive renal disease.
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- 2004
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6. Relaxin down-regulates renal fibroblast function and promotes matrix remodelling in vitro
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Tim D. Hewitson, Laura J. Parry, Kristen J. Kelynack, Gavin J. Becker, Rosemary Masterson, Ian A. Darby, Ross A. D. Bathgate, and Marina Martic
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Male ,medicine.medical_specialty ,Kidney Cortex ,Cell Culture Techniques ,Down-Regulation ,Biology ,Immediate early protein ,Immediate-Early Proteins ,Rats, Sprague-Dawley ,Fibrosis ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Fibroblast ,Relaxin ,Transplantation ,Kidney ,urogenital system ,Connective Tissue Growth Factor ,Fibroblasts ,medicine.disease ,Extracellular Matrix ,Rats ,CTGF ,Endocrinology ,medicine.anatomical_structure ,Nephrology ,Collagenase ,Intercellular Signaling Peptides and Proteins ,Procollagen ,hormones, hormone substitutes, and hormone antagonists ,medicine.drug ,Transforming growth factor - Abstract
Background. Renal fibroblasts are important effector cells in tubulointerstitial fibrosis, with experimental antifibrotic strategies focusing on the functional downregulation of these cells. Several experimental models of fibrosis have provided evidence for the effectiveness of the polypeptide hormone relaxin as a potential antifibrotic agent. This study was conducted to further elucidate the antifibrotic mechanisms of relaxin on renal fibroblasts in vitro. Methods. Rat cortical fibroblasts were obtained from outgrowth culture of renal tissue isolated from kidneys 3 days post-unilateral ureteric obstruction and constituted 100% of cells studied. A relaxin radio-receptor assay was used to establish binding of relaxin to renal fibroblasts in vitro. Functional studies then examined the effects of H2 relaxin (0, 1, 10 and 100 ng/ml) on fibroblast kinetics, expression of alpha-smooth muscle actin (-SMA), total collagen synthesis, collagenase production and collagen-I lattice contraction. CTGF mRNA expression was also measured by northern analysis. Results. H2 relaxin bound with high affinity to rat renal fibroblasts, but receptor numbers were low. Consistent with its previously reported bimodal effect, transforming growth factor (TGF-1) reduced fibroblast proliferation, an effect abrogated by H2 relaxin. Fibroblasts exposed to H2 relaxin (100 ng/ml) for 24 h demonstrated decreased immunostaining for -SMA and reduced -SMA protein expression compared with controls. There was a trend for a relaxin-mediated reduction in total collagen synthesis and 1(I) mRNA expression with large dose-related increases in collagenase protein expression being observed. TGF-1-stimulated collagen-I lattice contraction was significantly inhibited following co-incubation with 100 ng/ml relaxin. Incremental doses of H2 relaxin had no significant effect on CTGF mRNA expression. Conclusions. The findings of this study suggest that the antifibrotic effects of relaxin involve down-regulation of fibroblast activity, increase in collagenase synthesis and restructuring of collagen-I lattices, which are consistent with its known physiological role of matrix remodelling. Although there appears to be an interaction between TGF-1 and H2 relaxin, this does not appear to involve a reduction in CTGF mRNA expression.
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- 2004
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7. Parathyroid Hormone Has a Prosclerotic Effect on Vascular Smooth Muscle Cells
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Marina Martic, Melanie G. Tait, Kristen J. Kelynack, Vlado Perkovic, Gavin J. Becker, and Tim D. Hewitson
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medicine.medical_specialty ,Contraction (grammar) ,Vascular smooth muscle ,Cell division ,Arteriosclerosis ,Parathyroid hormone ,In Vitro Techniques ,Muscle, Smooth, Vascular ,Pathogenesis ,Fibrosis ,Internal medicine ,medicine ,Humans ,In patient ,skin and connective tissue diseases ,Aorta ,Cells, Cultured ,Uremia ,Accelerated atherosclerosis ,business.industry ,General Medicine ,medicine.disease ,Peptide Fragments ,Extracellular Matrix ,Endocrinology ,Parathyroid Hormone ,Nephrology ,sense organs ,Cardiology and Cardiovascular Medicine ,business ,Cell Division ,hormones, hormone substitutes, and hormone antagonists - Abstract
Although accelerated atherosclerosis and arteriosclerosis are common in patients with renal failure, the pathogenesis of these changes is poorly understood. Parathyroid hormone (PTH) levels are elevated in renal failure, and have been linked to uraemic vascular changes in some studies. We examined the in vitro effects of increasing doses of the 1–34 fragment of PTH on human aortic vascular smooth muscle cells (VSMCs). Factors examined were: (1) collagen production using tritiated hydroxyproline incorporation and transcription of procollagen α1(I) mRNA; (2) change in the surface area of collagen I lattices; (3) mRNA transcription of the collagen binding protein β1 integrin; (4) proliferation using tritiated thymidine incorporation, and (5) methyl tetrazolium salt conversion to estimate live cell number after 5 days’ exposure to PTH. PTH at a concentration of 200 pmol/l increased total collagen synthesis (188 ± 25% of control, p < 0.01) as well as transcription of procollagen α1(I) mRNA (136 ± 11% of control, p < 0.005). PTH also increased reorganisation of collagen I lattices (surface area 47 ± 8% of well for control vs. 35.7 ± 2.5 and 34.3 ± 3.0% for PTH 100 and 200 pmol/l, respectively, p = 0.02) and upregulated β1 integrin mRNA expression (160 ± 20% of control at PTH concentration of 200 pmol/l, p < 0.05). PTH had no effect on VSMC proliferation or number at doses up to 200 pmol/l. In conclusion, PTH increases production and reorganisation of collagen by VSMCs in vitro. It is possible that more aggressive control of hyperparathyroidism in patients with renal failure may help to reduce the burden of cardiovascular disease in this patient population.
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- 2003
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8. Contents Vol. 91, 2002
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Steven McTaggart, Hisaya Tada, D. Ljubanovic, Gunilla Halldén, M.J. Nubé, Marina Martic, Yasufumi Kyuden, Luisa Murer, Vincenzo Panichi, W.E.M. Schouten, Eugênio Pacelle Queiroz Madeira, Mariko Miyazaki, Yasuhiko Ueda, Hidetake Kurihara, Maria Norpoth, Guilherme Santoro-Lopes, Mutsuko Hidaka, Galal Mohamed Amer, Ali Moshfegh, Carlo Catalano, Chien-Liang Chen, Stefano De Pietro, Koji Kuboki, Ryoko Shimizu-Hirota, P. Hausfater, Susana Sampaio, Yoshinobu Fuke, Manuel Palacín, Luca Giovannini, Savvas Kazoulis, Satoshi Ogata, A.J. van Houte, Ikuo Horigome, Graziella Zacchello, Alberto Bettinelli, Batya Kristal, Tim D. Hewitson, Isao Shirato, Noriaki Yorioka, Makoto Watanabe, Renza Cristofani, Sérgio Fernando Ferreira Santos, Takao Kawamura, Y. Akai, S. Krizanac, Jacek Borawski, Maura Zanolari Calderari, Nobuoki Kohno, Marcella Normanno, Giulietta Sbragia, Iwan Setiawan, U. Assogba, Mitsuhiro Satoh, Mutsuo Taiji, Y. Asano, Mahmoud El-Baz, Daisuke Suzuki, Gavin J. Becker, Akira Saito, Giovanni Montini, Stefan Bröer, Yuji Fujita, Revital Shurtz-Swirski, Paul F. Laflam, P A Conz, Ching-Bun Chen, Ioannis Christoulakis, Toshihiro Shinosaki, Takafumi Ito, Takashi Uzu, Yasuhiro Akai, Kang-Ju Chou, G. Kantarcı, Shan Lin, Atsushi Tsuchida, Hideo Shiiki, Mari Kuroda, Atsushi Satomura, Huiqi Qu, Soichi Haraguchi, Michał Myśliwiec, Masayuki Iwano, Masafumi Yamato, M. Vrkljan, Petros Hatzilias, Yoshihiko Taniguchi, Atsushi Yamauchi, Emanuela Rizzioli, Mingcai Qiu, Mohamed Abel-Kader Sobh, Sana Sayed Gazarin, Shigemi Chiba, M. Schoorl, E. Kusano, Yukiteru Asakimori, Anna Maria Bianchi, Birgit Kallinowski, Mario G. Bianchetti, Armando A. Mendes, Yutaka Yaguchi, Isao Ishikawa, Manuel Pestana, Pia Thylén, M. Inoue, Shifra Sela, Po-Tsang Lee, Massimiliano Migliori, Jean-Pierre Guignard, Olaf Hergesell, P. Lebon, Hideto Sakai, Yee-Hsuan Chiou, Kristen J. Kelynack, Alberto A.E. Bertelli, Osamu Hotta, Shaul M. Shasha, Tomokazu Nagano, Lambertus Arisz, Martin Zeier, H. Furuya, Kefaia El-Sayed Mohamed, A. Bihorac, Ç. Özener, Lara Alonso da Silva, Hiroshi Noguchi, Toshio Miyata, Takao Saruta, E. Akoğlu, Stefan H. Jacobson, Krystyna Pawlak, Ivano Moschèn, Maria Rita Metelli, Hsiao-Min Chung, Takashi Furuta, Adriana MacIntyre Innocenzi, Yoshiharu Nishitani, Kazumasa Hamano, Manabu Asano, Mizuo Mifune, Kayoko Nomura, Tatsuo Kobayashi, Gian Marco Ghiggeri, João Ramos, Mahmoud Wageh Rasem, José Gerardo Oliveira, Hassan Abd-El-Hady Ahmed, Hiroyuki Ohi, S. Tokay, Matsuhiko Hayashi, Osama Gheith, Carla Carasi, P. Xavier, Ikue Mori-Kudo, Hiroo Noshiro, Kazuhiko Funabiki, Michael Nomikos, Chizuru Ishiguro, Ikuko Miyai, Maria Magalhães, Péter Tóth-Heyn, Satoko Honda, Manuela Della Vella, Daniele Taccola, Gianluca Caridi, Yasuharu Nomura, Hideyuki Kurioka, Joachim Lundahl, P. Cacoub, Y. Ando, Ronit Geron, Toshiki Inokuchi, Eduardo H. Garin, Galina Shapiro, Roberto Palla, Shih-Yuan Hung, Yasuhiko Tomino, Ciro Tetta, Surinder Cheema-Dhadli, Alexandra Albers, Sarantos Xirakis, Florian Lang, Mitchell L. Halperin, Hideaki Nakaya, Ken Takahara, Fabio Fabbian, Jürgen Floege, Yoshio Taguma, Koji Harada, Hua-Chang Fang, F. A. W. Kemperman, Satoshi Horikoshi, Rodo O. von Vigier, Morito Endo, Omar da Rosa Santos, Norio Sunagawa, M.P.C. Grooteman, Fatma Elhusseini, Takayuki Fujita, Raymond T. Krediet, J.C. Piette, Rajiv Kumar, Hiroyuki Sasamura, K. Tabei, Ioannis Tzanakis, K. Galesic, Mie Ko, and Isao Ohsawa
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Traditional medicine ,business.industry ,Medicine ,business - Published
- 2002
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9. Subject Index Vol. 91, 2002
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Hiroyuki Sasamura, Olaf Hergesell, K. Tabei, Osamu Hotta, Norio Sunagawa, Ioannis Tzanakis, Emanuela Rizzioli, Mingcai Qiu, Paul F. Laflam, Gian Marco Ghiggeri, Yasufumi Kyuden, D. Ljubanovic, M.J. Nubé, W.E.M. Schouten, Carla Carasi, Joachim Lundahl, Mariko Miyazaki, Isao Ishikawa, Yasuhiko Ueda, Kazuhiko Funabiki, Michael Nomikos, Tatsuo Kobayashi, Maria Magalhães, Manabu Asano, Takayuki Fujita, Shaul M. Shasha, Revital Shurtz-Swirski, Susana Sampaio, H. Furuya, Kefaia El-Sayed Mohamed, Mitsuhiro Satoh, A. Bihorac, Ç. Özener, Renza Cristofani, Ikue Mori-Kudo, U. Assogba, Satoshi Ogata, A.J. van Houte, Shifra Sela, Hidetake Kurihara, Maria Rita Metelli, Kayoko Nomura, Giovanni Montini, Atsushi Satomura, S. Krizanac, Rodo O. von Vigier, Morito Endo, Jürgen Floege, Lara Alonso da Silva, Yasuharu Nomura, Satoko Honda, Manuel Pestana, Hassan Abd-El-Hady Ahmed, Omar da Rosa Santos, P. Cacoub, Toshihiro Shinosaki, Ikuko Miyai, Y. Asano, Yoshiharu Nishitani, Yoshio Taguma, Chien-Liang Chen, Chizuru Ishiguro, P. Xavier, Gianluca Caridi, Marina Martic, Jean-Pierre Guignard, Maura Zanolari Calderari, Akira Saito, Hua-Chang Fang, F. A. W. Kemperman, Huiqi Qu, Atsushi Yamauchi, Hisaya Tada, Gavin J. Becker, Koji Harada, Mohamed Abel-Kader Sobh, Galal Mohamed Amer, Luisa Murer, P A Conz, Hiroyuki Ohi, Ali Moshfegh, Armando A. Mendes, Osama Gheith, Daisuke Suzuki, Fatma Elhusseini, Ciro Tetta, Eugênio Pacelle Queiroz Madeira, Yoshinobu Fuke, Takashi Uzu, Kang-Ju Chou, Savvas Kazoulis, Manuel Palacín, Ioannis Christoulakis, P. Hausfater, G. Kantarcı, Hideyuki Kurioka, Ivano Moschèn, Stefan H. Jacobson, Graziella Zacchello, Satoshi Horikoshi, Atsushi Tsuchida, Ryoko Shimizu-Hirota, Petros Hatzilias, Alberto Bettinelli, Tim D. Hewitson, Raymond T. Krediet, J.C. Piette, Jacek Borawski, S. Tokay, Takafumi Ito, Mutsuo Taiji, Ronit Geron, Makoto Watanabe, M. Schoorl, Carlo Catalano, Pia Thylén, Péter Tóth-Heyn, Ching-Bun Chen, Rajiv Kumar, Shigemi Chiba, Stefan Bröer, Takao Saruta, Mahmoud El-Baz, Yuji Fujita, E. Akoğlu, Roberto Palla, Michał Myśliwiec, Toshiki Inokuchi, Yasuhiko Tomino, Yasuhiro Akai, E. Kusano, Yukiteru Asakimori, Maria Norpoth, Shih-Yuan Hung, Yee-Hsuan Chiou, Kristen J. Kelynack, Hideo Shiiki, Alberto A.E. Bertelli, Mari Kuroda, Yoshihiko Taniguchi, Kazumasa Hamano, Massimiliano Migliori, Steven McTaggart, P. Lebon, Toshio Miyata, Vincenzo Panichi, Gunilla Halldén, Soichi Haraguchi, Mario G. Bianchetti, Po-Tsang Lee, M.P.C. Grooteman, Hideaki Nakaya, Ken Takahara, Fabio Fabbian, Yutaka Yaguchi, Florian Lang, Anna Maria Bianchi, Birgit Kallinowski, Mitchell L. Halperin, M. Inoue, Mizuo Mifune, Masayuki Iwano, Masafumi Yamato, Stefano De Pietro, Hiroshi Noguchi, Koji Kuboki, Hsiao-Min Chung, Hideto Sakai, Tomokazu Nagano, Iwan Setiawan, Surinder Cheema-Dhadli, Nobuoki Kohno, Shan Lin, M. Vrkljan, Isao Shirato, Noriaki Yorioka, Adriana MacIntyre Innocenzi, Marcella Normanno, José Gerardo Oliveira, Matsuhiko Hayashi, Y. Ando, Lambertus Arisz, Krystyna Pawlak, Galina Shapiro, Ikuo Horigome, Takashi Furuta, Y. Akai, Batya Kristal, João Ramos, Mahmoud Wageh Rasem, Alexandra Albers, Sarantos Xirakis, Sérgio Fernando Ferreira Santos, Takao Kawamura, Martin Zeier, Hiroo Noshiro, Manuela Della Vella, Daniele Taccola, Isao Ohsawa, K. Galesic, Mie Ko, Sana Sayed Gazarin, Eduardo H. Garin, Guilherme Santoro-Lopes, Mutsuko Hidaka, Luca Giovannini, and Giulietta Sbragia
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Index (economics) ,business.industry ,Statistics ,Medicine ,Subject (documents) ,business - Published
- 2002
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10. Human renal fibroblast contraction of collagen I lattices is an integrin‐mediated process
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Ian A. Darby, Gavin J. Becker, Tim D. Hewitson, Kristen J. Kelynack, and Kathy Nicholls
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Adult ,Male ,Integrins ,Pathology ,medicine.medical_specialty ,Kidney Cortex ,Receptors, Collagen ,Integrin ,Cell Culture Techniques ,Biology ,Integrin alpha1beta1 ,Collagen receptor ,Extracellular matrix ,Receptors, Fibronectin ,medicine ,Humans ,Fibroblast ,Cells, Cultured ,Aged ,Transplantation ,Integrin beta1 ,Integrin alpha3beta1 ,Fibroblasts ,Middle Aged ,Cell biology ,Fibronectin ,Collagen, type I, alpha 1 ,medicine.anatomical_structure ,Nephrology ,Alpha-5 beta-1 ,biology.protein ,Female ,Collagen - Abstract
BACKGROUND: Expression of the beta1 family of integrins allows dermal fibroblasts in wounds to contribute to the healing process through migration, adhesion, synthesis, and rearrangement of extracellular matrix. To date the ability of human renal fibroblasts to reorganize collagens and the role of cell surface receptors in this process remain unknown. METHODS: Renal fibroblasts were grown from the cortical tissue of surgically removed human kidneys. The ability of human renal fibroblasts to reorganize interstitial collagen I was examined in vitro using solidified collagen I lattices. Integrin function was blocked by incubating fibroblasts with isotype-specific antibodies prior to addition to collagen I lattices. RESULTS: Human renal fibroblasts embedded in collagen I lattices progressively decreased lattice diameter to 60.6+/-11.4% of initial diameter at 48 h post-release (P
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- 2000
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11. Pentoxifylline Reduces in vitro Renal Myofibroblast Proliferation and Collagen Secretion
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Kristen J. Kelynack, Marina Martic, Gavin J. Becker, Eugenia Pedagogos, and Tim D. Hewitson
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medicine.medical_specialty ,Phosphodiesterase Inhibitors ,In Vitro Techniques ,Biology ,Kidney ,Pentoxifylline ,Rats, Sprague-Dawley ,Internal medicine ,medicine ,Animals ,Cells, Cultured ,Dose-Response Relationship, Drug ,Cell growth ,Muscle, Smooth ,Transforming growth factor beta ,Fibroblasts ,In vitro ,Rats ,medicine.anatomical_structure ,Endocrinology ,Nephrology ,Cell culture ,biology.protein ,Collagen ,Myofibroblast ,Cell Division ,medicine.drug ,Explant culture - Abstract
Interstitial myofibroblasts (MF) are cells with features of both smooth muscle cells and fibroblasts. They have been universally recognized in situations of tubulointerstitial injury, where their presence has been shown to be a marker of disease progression. The objective of this study was to determine if functions of MF relevant to fibrogenesis can be modified in vitro by the phosphodiesterase inhibitor pentoxifylline (PTX). MF were obtained from sub-culture of normal rat kidney explant outgrowths maintained in DMEM + 20% fetal calf serum (FCS), supplemented with antibiotics. Cells were characterized on the basis of growth characteristics and immunohistochemistry. MF constituted >95% of cells at passage 3. Cell culture media was supplemented with the potential antagonist PTX alone (0, 1, 10, 100 μg/ml) and in combination with TGFβ1 (5 ng/ml). Population kinetics, proliferation and collagen production were determined from cell growth, [3H]thymidine incorporation and [3H]proline incorporation in collagenous proteins, respectively. Both serum-stimulated population growth and proliferation were reduced in a linear fashion by 1, 10 and 100 μg/ml PTX (all p < 0.05 versus 0 μg/ml). Effect of PTX on cell population growth was however reversible when PTX was removed. Basal collagen secretion was decreased by PTX at 10 and 100 μg/ml (p < 0.05 versus 0 μg/ml), although cell layer collagen remained unchanged. Collagen production (secreted and cell layer) was augmented by 5 ng/ml TGFβ1. These effects on collagen production were partially reduced when 100 μg/ml PTX was added. The authors conclude that myofibroblast function can be altered with agonists/antagonists. Attempts to down-regulate fibrogenic functions of MF may therefore offer a valuable therapeutic strategy.
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- 2000
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12. Contents Vol. 19, 1999
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Tempie E. Hulbert-Shearon, Roxiana Sadikot, Michael Borucki, Yoshiko Hayashi, Nagaraja Rao Sridhar, Noboru Kishimoto, Nobuyuki Miyatake, Masahiko Tozawa, Seong Wook Park, Lawrence Y. Agodoa, Shinichro Yoshi, Tejinder S. Ahuja, Robert A. Wolfe, José J. Escarce, Dewey Butts, Kunitoshi Iseki, Koshiro Fukiyama, Harold I. Feldman, Edward Greeno, Chiho Iseki, Akinlolu O. Ojo, Tim D. Hewitson, Michael Hollander, W. Brian Reeves, Robert O. Berkseth, Shuzou Gomikawa, Lionel Rostaing, Marie-Hélène Chabannier, Zensuke Ota, Masahiko Kushiro, Kostas C. Siamopoulos, Julie A. Hanson, Jae Young Kang, Anne Rouzaud, Mary Jo Shaver, Kenichi Shikata, Srinivasan Rajaraman, Jean-Marc Cisterne, David C. Dahl, Romesh Kohli, Soon Bae Kim, Friedrich K. Port, Dominique Durand, Kazuhiko Suzuki, Warren B. Bilker, Moses Elisaf, Kathleen Ferrand, Jean Tkaczuk, Jee Hyun Park, Kulwant S. Modi, Kazue Hironaka, Kathy Nicholls, Yoshihiro Takamitu, Patrick Hayes, Juan P. Bosch, Amy M. Smith, Hirofumi Makino, Susie Q. Lew, Maria P. Varela, Kristen J. Kelynack, Osamu Morita, Anne Modesto, Seung-Jung Park, Christopher W. Simmons, Won Seok Yang, Leah Pinnavaia, Saeko Ogawa, Jung Sik Park, Marjorie Funtanilla, Gavin J. Becker, Eugenia Pedagogos, John H. Holmes, Kosuke Ota, Vahakn B. Shahinian, Ronald Schut, Rachel L. Whyte, George Papandenatos, Mark V. Pauly, John T. Daugirdas, Monica Hackett, and Deborah Reger
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Traditional medicine ,Nephrology ,business.industry ,Medicine ,business - Published
- 1999
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13. Renal Myofibroblasts Contract Collagen I Matrix Lattices in vitro
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Kathy Nicholls, Eugenia Pedagogos, Kristen J. Kelynack, Tim D. Hewitson, and Gavin J. Becker
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Male ,Collagen i ,Pathology ,medicine.medical_specialty ,Contraction (grammar) ,Cell ,Biology ,Kidney ,Rats, Sprague-Dawley ,Smooth muscle ,medicine ,Animals ,Fibroblast ,Cells, Cultured ,Wound Healing ,Microscopy, Confocal ,Integrin beta1 ,Muscle, Smooth ,Fibroblasts ,In vitro ,Extracellular Matrix ,Rats ,medicine.anatomical_structure ,Nephrology ,Keratins ,Collagen ,Myofibroblast - Abstract
Myofibroblasts, cells with both fibroblastic and smooth muscle cell features, have been implicated in renal scarring. In addition to synthetic properties, contractile features and integrin expression may allow myofibroblasts to rearrange and contract interstitial collagenous proteins. Myofibroblasts from normal rat kidneys were grown in cell-populated collagen lattices to measure cell generated contraction. Following detachment of cell populated collagen lattices, myofibroblasts progressively contracted collagen lattices, reducing lattice diameter by 42% at 24 h. Alignment of myofibroblasts, rearrangement of fibrils and β1 integrin expression were observed within lattices. We postulate that interstitial myofibroblasts contribute to renal scarring through manipulation of collagenous proteins.
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- 1999
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14. Cell kinetics and tissue contraction following renal parenchymal cell death
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Colin L. Jones, Kristen J. Kelynack, Gavin J. Becker, Kathy Nicholls, and Tim D. Hewitson
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Kidney ,Pathology ,medicine.medical_specialty ,business.industry ,Monocyte ,General Medicine ,medicine.disease ,Lesion ,Extracellular matrix ,medicine.anatomical_structure ,Nephrology ,Parenchyma ,medicine ,medicine.symptom ,Fibroblast ,business ,Myofibroblast ,Infiltration (medical) - Abstract
SUMMARY: The kinetics of interstitial extracellular matrix remodelling in the kidney following parenchymal cell death were studied after injury was produced by a single application of liquid nitrogen to the surface of the kidney. the size of the injury decreased to 30% of the original size at 4 days and to 5% at 16 days after injury. the primary response to injury consisted of an acute polymorphonuclear infiltration followed by monocyte/macrophage and myofibroblast accumulation from day 2 to 8. Collagen III accumulation increased progressively until day 16 and was inversely related to lesion size. These results demonstrate the time course of cellular events and collagen synthesis in renal tissue remodelling following renal parenchymal cell death.
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- 1998
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15. Explanting Is an Ex Vivo Model of Renal Epithelial-Mesenchymal Transition
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Kristen J. Kelynack, Tim D. Hewitson, Dodie S. Pouniotis, Gavin J. Becker, Catherine E. Winbanks, and Ian A. Darby
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Pathology ,medicine.medical_specialty ,Epithelial-Mesenchymal Transition ,Article Subject ,lcsh:Biotechnology ,Health, Toxicology and Mutagenesis ,Cell Culture Techniques ,lcsh:Medicine ,Vimentin ,macromolecular substances ,Kidney ,Kidney Tubules, Proximal ,Rats, Sprague-Dawley ,Cytokeratin ,lcsh:TP248.13-248.65 ,Genetics ,medicine ,Animals ,Epithelial–mesenchymal transition ,Myofibroblasts ,Molecular Biology ,Cell Proliferation ,Staining and Labeling ,biology ,Immunochemistry ,lcsh:R ,Endothelial Cells ,Cell Differentiation ,Epithelial Cells ,Mesenchymal Stem Cells ,General Medicine ,Molecular biology ,Rats ,Phenotype ,medicine.anatomical_structure ,Tubulointerstitial fibrosis ,biology.protein ,Molecular Medicine ,Desmin ,Myofibroblast ,Biomarkers ,Ex vivo ,Research Article ,Biotechnology - Abstract
Recognised by their de novo expression of alpha-smooth muscle actin (SMA), recruitment of myofibroblasts is key to the pathogenesis of fibrosis in chronic kidney disease. Increasingly, we realise that epithelial-mesenchymal transition (EMT) may be an important source of these cells. In this study we describe a novel model of renal EMT. Rat kidney explants were finely diced on gelatin-coated Petri dishes and cultured in serum-supplemented media. Morphology and immunocytochemistry were used to identify mesenchymal (vimentin+, α-smooth muscle actin (SMA)+, desmin+), epithelial (cytokeratin+), and endothelial (RECA+) cells at various time points. Cell outgrowths were all epithelial in origin (cytokeratin+) at day 3. By day 10, 50 ± 12% (mean ± SE) of cytokeratin+ cells double-labelled for SMA, indicating EMT. Lectin staining established a proximal tubule origin. By day 17, cultures consisted only of myofibroblasts (SMA+/cytokeratin−). Explanting is a reproducible ex vivo model of EMT. The ability to modify this change in phenotype provides a useful tool to study the regulation and mechanisms of renal tubulointerstitial fibrosis.
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- 2011
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16. Cell-Populated Floating Collagen Lattices: An In Vitro Model of Parenchymal Contraction
- Author
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Kristen J. Kelynack
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Contraction (grammar) ,Chemistry ,Mesenchymal stem cell ,Cell ,Glomerulosclerosis ,Anatomy ,medicine.disease ,In vitro ,Cell biology ,Extracellular matrix ,medicine.anatomical_structure ,Parenchyma ,medicine ,Tubulointerstitial fibrosis - Abstract
The pathology of progressive renal disease is characterized by glomerular and interstitial inflammation, glomerulosclerosis, and tubulointerstitial fibrosis. This is a consequence of excessive matrix synthesis, reduced matrix degradation, and contraction (reorganization) of extracellular matrix. Fibroblasts, and to a lesser degree, other mesenchymal cells, are known to contribute to renal scar formation through local proliferation, synthesis, and reorganization of matrix proteins. Although much work has focused on the balance between collagen synthesis and degradation, the mechanisms of parenchymal collapse and contraction are becoming increasingly important. Like their counterparts in the skin, the contractile properties of renal fibroblasts are now well recognized. This chapter details an in vitro method for studying the contraction of collagens by homogeneous populations of cultured cells. The method can be altered so that reagents influencing this process may also be studied.
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- 2008
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17. Histochemical Localization of Cell Proliferation Using In Situ Hybridization for Histone mRNA
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Ian A. Darby, Kristen J. Kelynack, and Tim D. Hewitson
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biology ,medicine.drug_class ,Cell growth ,In situ hybridization ,Cell cycle ,Monoclonal antibody ,Molecular biology ,Cell biology ,chemistry.chemical_compound ,Histone ,chemistry ,In vivo ,biology.protein ,medicine ,Thymidine ,Bromodeoxyuridine - Abstract
Monoclonal antibodies to proliferation associated antigens have long been used to histologically localize mitogenesis. However, techniques that distinguish cells in the synthetic or S phase have tended to rely on the in vivo incorporation of tritiated thymidine or thymidine analogs such as bromodeoxyuridine. The necessity to pulse with these labels before retrieving tissue means that they cannot be used in humans and are not available retrospectively. Measuring expression of histones serves as a useful adjunct to these techniques. As expression of histone proteins (H2A, H2B, H3, H4) are restricted to the synthetic phase of the cell cycle, hybridization for histone mRNA precisely distinguishes those cells in the S phase. Measuring their expression can easily be applied to the histological localization of proliferation, and can be used both prospectively and with archived tissue specimens. Several histone in situ hybridization probes and nonradioactive detection systems are now available commercially. A generalized protocol for their use in measuring in situ proliferation is provided in this chapter.
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- 2006
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18. Histochemical localization of cell proliferation using in situ hybridization for histone mRNA
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Tim D, Hewitson, Kristen J, Kelynack, and Ian A, Darby
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Histones ,Histocytochemistry ,Animals ,Humans ,RNA, Messenger ,In Situ Hybridization ,Cell Proliferation ,S Phase - Abstract
Monoclonal antibodies to proliferation associated antigens have long been used to histologically localize mitogenesis. However, techniques that distinguish cells in the synthetic or S phase have tended to rely on the in vivo incorporation of tritiated thymidine or thymidine analogs such as bromodeoxyuridine. The necessity to pulse with these labels before retrieving tissue means that they cannot be used in humans and are not available retrospectively. Measuring expression of histones serves as a useful adjunct to these techniques. As expression of histone proteins (H2A, H2B, H3, H4) are restricted to the synthetic phase of the cell cycle, hybridization for histone mRNA precisely distinguishes those cells in the S phase. Measuring their expression can easily be applied to the histological localization of proliferation, and can be used both prospectively and with archived tissue specimens. Several histone in situ hybridization probes and nonradioactive detection systems are now available commercially. A generalized protocol for their use in measuring in situ proliferation is provided in this chapter.
- Published
- 2006
19. Thrombin is a pro-fibrotic factor for rat renal fibroblasts in vitro
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G J Becker, Marina Martic, Eleanor J. Mackie, Charles N. Pagel, Kristen J. Kelynack, and Tim D. Hewitson
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Male ,Serine Proteinase Inhibitors ,Physiology ,Myocytes, Smooth Muscle ,Biology ,Fibrinogen ,Kidney ,Fibrin ,Amino Acid Chloromethyl Ketones ,Rats, Sprague-Dawley ,Thrombin ,Fibrosis ,Genetics ,medicine ,Animals ,Receptor, PAR-1 ,RNA, Messenger ,Fibroblast ,Cells, Cultured ,Kidney metabolism ,General Medicine ,Fibroblasts ,medicine.disease ,Molecular biology ,In vitro ,Actins ,Rats ,medicine.anatomical_structure ,Nephrology ,Cancer research ,biology.protein ,Collagen ,Gels ,Biomarkers ,Cell Division ,circulatory and respiratory physiology ,medicine.drug - Abstract
Background: Generation of thrombin occurs in response to parenchymal injury. Thrombin not only converts plasma fibrinogen into an insoluble fibrin clot, but also potentially augments inflammation through receptor-mediated activity. This study examines whether thrombin may potentially exacerbate fibrosis by upregulating the function of interstitial fibroblasts in vitro. Methods: Fibroblasts were isolated by explant outgrowth culture of rat kidneys. Subcultured cells were grown in DMEM+10% FCS supplemented with 0.1–0.5 U/ml thrombin. Functional parameters examined included kinetics (thymidine incorporation and change in cell number), differentiation (Western blotting for α-smooth muscle actin; αSMA), expression of procollagen α1(I) (Northern blotting) and contraction of collagen I lattices. RT-PCR was used to characterise expression of protease-activated receptors (PAR) previously implicated in thrombin’s cellular effects. Results: Cell population growth was increased 66 ± 41 and 47 ± 41% by 0.1 and 0.5 U/ml thrombin respectively (both p < 0.05 vs. basal). Likewise, 0.5 U/ml thrombin increased corrected procollagen α1(I) expression 2.4-fold (p < 0.05 vs. basal) and exacerbated the ability of fibroblasts to contract collagen matrix (p < 0.05 vs. basal). These effects were not associated with any change in expression of the myofibroblast marker αSMA. Effects on cell number were inhibited by treatment with (D)-Phe-Pro-Arg-chloromethylketone HCl (PPACK) suggesting that functional effects were mediated by serine protease activity. PAR-1 was the only fully functional known thrombin receptor expressed by these cells. Conclusion: Thrombin is a potential unrecognised fibroblast agonist in renal disease. Further studies of thrombin and its receptors may yield valuable insights into the pathogenesis of interstitial fibrosis.
- Published
- 2003
20. Dipyridamole inhibits in vitro renal fibroblast proliferation and collagen synthesis
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Marina Martic, Melanie G. Tait, Gavin J. Becker, Tim D. Hewitson, and Kristen J. Kelynack
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Male ,medicine.medical_specialty ,Indoles ,Cell Survival ,Phosphodiesterase Inhibitors ,Biology ,Kidney ,Nitric Oxide ,Pathology and Forensic Medicine ,Immunoenzyme Techniques ,Maleimides ,Rats, Sprague-Dawley ,chemistry.chemical_compound ,Fibrosis ,Internal medicine ,medicine ,Animals ,RNA, Messenger ,Fibroblast ,Fluorescein isothiocyanate ,Fluorescent Antibody Technique, Indirect ,Cells, Cultured ,In Situ Hybridization ,Sulfonamides ,Cell growth ,General Medicine ,Dipyridamole ,Fibroblasts ,medicine.disease ,Isoquinolines ,Molecular biology ,In vitro ,Rats ,Procollagen peptidase ,Disease Models, Animal ,medicine.anatomical_structure ,Endocrinology ,chemistry ,Cell culture ,Collagen ,Cell Division ,medicine.drug ,Ureteral Obstruction - Abstract
Fibroblasts are universally recognized in situations of tubulointerstitial injury, where their presence has been shown to be a marker of disease progression. The objective of this study was to determine whether the functions of fibroblasts relevant to fibrogenesis can be modified in vitro with dipyridamole. Cells were obtained from obstructed rat renal tissue and characterized on the basis of immunohistochemical findings. Fibroblasts constituted all of the cells from passage 3. Functional parameters were measured in cells cultured with 1, 5, and 50 micromol/L dipyridamole and compared to basal parameters of cells grown in Dulbecco's modified Eagle's medium plus 10% fetal calf serum (control). Northern-blot analysis indicated that dipyridamole decreased procollagen alpha1(I) messenger ribonucleic acid expression (P.05, 50 micromol/L vs control), results that were reflected in a reduction in total collagen secretion as measured on the basis of hydroxyproline incorporation (P.001, 50 micromol/L vs control). Mitogenesis, as measured on the basis of incorporation of tritiated thymidine, was decreased in a dose-dependent fashion by dipyridamole. Likewise, 50 micromol/L dipyridamole reduced cell-population growth to 16.8% +/- 2.1% of basal growth over 3 days (P.001 vs control). Effects of dipyridamole on population growth were prevented by coincubation with a protein kinase G inhibitor peptide (P.001 vs 50 micromol/L dipyridamole; P = not significant vs control). No such effect was observed with inhibitors for protein kinase A (H-89) and protein kinase C (bisindolylmaleimide I). Consistent with a protein kinase G-dependent mechanism, immunofluorescence staining indicated that dipyridamole increased basal expression of the inducible form of nitric oxide synthase. In conclusion, the results of this study demonstrate that at clinically relevant concentrations, dipyridamole inhibits profibrotic activities of renal fibroblasts. Effects on mitogenesis are mediated through a cyclic guanosine monophosphate-protein kinase G effector pathway.
- Published
- 2002
21. Lovastatin downregulates renal myofibroblast function in vitro
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Gavin J. Becker, Tim D. Hewitson, Kristen J. Kelynack, Steven McTaggart, and Marina Martic
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Male ,Pathology ,medicine.medical_specialty ,Interstitial nephritis ,Down-Regulation ,macromolecular substances ,Kidney ,Collagen Type I ,Extracellular matrix ,Rats, Sprague-Dawley ,medicine ,Animals ,Lovastatin ,Fibroblast ,Cells, Cultured ,biology ,Dose-Response Relationship, Drug ,business.industry ,Cell Cycle ,Fibroblasts ,medicine.disease ,Hydroxymethylglutaryl-CoA reductase ,Immunohistochemistry ,Matrix Metalloproteinases ,Rats ,medicine.anatomical_structure ,Phenotype ,HMG-CoA reductase ,biology.protein ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,business ,Myofibroblast ,medicine.drug ,Kidney disease - Abstract
Interstitial fibrosis is recognised as the best histological predictor of progressive renal disease. Myofibroblasts contribute to this process through several functions including hyperproliferation, collagen and collagenase synthesis and reorganisation of extracellular matrix. Recent limited in vitro studies suggest that 3-hydroxy-3-methylglutaryl-coenzyme A (HMG CoA) reductase inhibitors may reduce renal injury not only through their lipid-lowering effects but also by antagonising myofibroblast function. This study therefore examined the effects of lovastatin on the above interstitial myofibroblast behaviours in vitro. Primary cultures of rat renal cortical myofibroblasts were grown by explantation and characterised by immunohistochemistry. Dose response effects of lovastatin (0, 15, 30 µM) in DMEM and 10% FCS were examined on myofibroblast kinetics, total collagen synthesis, collagen I lattice contraction and actin filament rearrangement. Lovastatin decreased myofibroblast proliferation and growth. Likewise, collagen I lattice contraction and actin filament rearrangement were partially inhibited when lovastatin was added at 30 µM. In addition, lovastatin decreased both collagen and collagenase synthesis. Our results suggest that myofibroblast function may be downregulated by lovastatin in vitro. Although a decrease in myofibroblast activity may offer potential benefit in the prevention of progressive scarring, further studies will be necessary to determine the relative importance of these functions.
- Published
- 2002
22. LOVASTATIN DOWNREGULATES RENAL MYOFIBROBLAST FUNCTION IN VITRO
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Marina Martic, Steven McTaggart, Tim D. Hewitson, Vlado Perkovic, G J Becker, and Kristen J. Kelynack
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Nephrology ,business.industry ,medicine ,General Medicine ,Lovastatin ,Pharmacology ,business ,Myofibroblast ,Function (biology) ,In vitro ,medicine.drug - Published
- 2002
- Full Text
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23. Consultants for the American Journal of Nephrology 1999
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Ronald Schut, Lawrence Y. Agodoa, Robert A. Wolfe, Tim D. Hewitson, Michael Hollander, Masahiko Tozawa, Nobuyuki Miyatake, Monica Hackett, Romesh Kohli, Friedrich K. Port, Warren B. Bilker, Kosuke Ota, Kulwant S. Modi, Kazue Hironaka, Won Seok Yang, Leah Pinnavaia, Maria P. Varela, Deborah Reger, Dominique Durand, Kazuhiko Suzuki, Seong Wook Park, Chiho Iseki, Anne Modesto, José J. Escarce, W. Brian Reeves, Yoshiko Hayashi, Koshiro Fukiyama, Zensuke Ota, Tejinder S. Ahuja, Srinivasan Rajaraman, Edward Greeno, Eugenia Pedagogos, Harold I. Feldman, Lionel Rostaing, Kunitoshi Iseki, Dewey Butts, Juan P. Bosch, Masahiko Kushiro, Amy M. Smith, Patrick Hayes, Moses Elisaf, Anne Rouzaud, Susie Q. Lew, Marie-Hélène Chabannier, Jean Tkaczuk, Robert O. Berkseth, Kristen J. Kelynack, George Papandenatos, Jean-Marc Cisterne, Shuzou Gomikawa, Seung-Jung Park, Osamu Morita, Jae Young Kang, Gavin J. Becker, Kostas C. Siamopoulos, Shinichro Yoshi, Tempie E. Hulbert-Shearon, Marjorie Funtanilla, Vahakn B. Shahinian, Saeko Ogawa, David C. Dahl, Kathy Nicholls, Kathleen Ferrand, Kenichi Shikata, Jee Hyun Park, Noboru Kishimoto, Mark V. Pauly, Hirofumi Makino, Nagaraja Rao Sridhar, Michael Borucki, John H. Holmes, Christopher W. Simmons, Jung Sik Park, John T. Daugirdas, Yoshihiro Takamitu, Rachel L. Whyte, Julie A. Hanson, Mary Jo Shaver, Soon Bae Kim, Akinlolu O. Ojo, and Roxiana Sadikot
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Nephrology ,medicine.medical_specialty ,business.industry ,Internal medicine ,Family medicine ,medicine ,business - Published
- 1999
- Full Text
- View/download PDF
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