Relevant bibliographies by topics / Continuous cell line

Academic literature on the topic 'Continuous cell line'

Author: Grafiati
Published: 10 December 2022
Last updated: 28 January 2023

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Journal articles on the topic "Continuous cell line"

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Hiraga, H., Takayuki Nojima, Syuiti Abe, Katsushige Yamashiro, Shinya Yamawaki, Kiyoshi Kaneda, and Kazuo Nagashima. "Establishment of a new continuous clear cell sarcoma cell line." Virchows Archiv 431, no. 1 (July 4, 1997): 45–51. http://dx.doi.org/10.1007/s004280050068.

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Ward, G. B., T. J. Kelly, C. W. Woods, and E. P. Marks. "Ecdysteroid production by a continuous insect cell line." Archives of Insect Biochemistry and Physiology 5, no. 2 (June 1987): 91–98. http://dx.doi.org/10.1002/arch.940050204.

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KOMATSUBARA, Kazunori, Yuji SIMOGONYA, and Noriyuki KATAOKA. "Effect of long-term continuous rotational stimulation on osteoblast-like cell line." Proceedings of the JSME Conference on Frontiers in Bioengineering 2020.31 (2020): 2A22. http://dx.doi.org/10.1299/jsmebiofro.2020.31.2a22.

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Ammar, Ayman A., and Ahmed M. Wahab. "Development of a Continuous Cell Line from Tilapia Liver." Journal of Ecology of Health & Environment 7, no. 1 (January 1, 2019): 1–5. http://dx.doi.org/10.18576/jehe/070101.

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Sherwood, J. B., and D. Shouval. "Continuous production of erythropoietin by an established human renal carcinoma cell line: development of the cell line." Proceedings of the National Academy of Sciences 83, no. 1 (January 1, 1986): 165–69. http://dx.doi.org/10.1073/pnas.83.1.165.

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MITSUHASHI, JUN, and AKIKO SHOZAWA. "Continuous Cell Lines from Larval Hemocytes of the Cabbage Armyworm, Mamestra brassiae. (continuous cell line/hemocytes/Mamestra brassicae)." Development, Growth and Differentiation 27, no. 5 (October 1985): 599–606. http://dx.doi.org/10.1111/j.1440-169x.1985.00599.x.

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Shimomura, S., N. Komatsu, N. Frickhofen, S. Anderson, S. Kajigaya, and NS Young. "First continuous propagation of B19 parvovirus in a cell line." Blood 79, no. 1 (January 1, 1992): 18–24. http://dx.doi.org/10.1182/blood.v79.1.18.18.

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Abstract The pathogenic human parvovirus B19 has extreme tropism for human erythroid progenitor cells and has resisted cultivation in conventional cell lines. We report first propagation of this virus in an erythropoietin-dependent strain of a megakaryoblastic leukemia cell line called UT-7. Virus protein was present in about 5% of cells after 1 week of culture. Appropriate ratios of major and minor capsid proteins were determined by immunoblot, and newly synthesized capsid protein was detected by immunoprecipitation of radioactively labeled cell lysates. High molecular weight monomer and dimer intermediates were detected by Southern analysis, indicating active viral replication. Approximately 1,000 genome copies were present per infected cell, and at the optimal multiplicity of infection 20- to 50- fold more virus was produced than inoculated. Virus propagation only occurred in UT-7 cells that were adapted to growth in erythropoietin; virus signal was not detected in UT-7 cells adapted for growth in granulocyte-macrophage colony-stimulating factor or interleukin-3, even with exposure to erythropoietin for several days. Infectious virus was detected in cultures as long as 3 months after inoculation. Despite persistence, there was no evidence of viral integration on Southern analysis. This cell line may prove useful for the production of infectious virus and in the analysis of B19 parvovirus persistence, cytotoxicity, and permissivity.
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Shimomura, S., N. Komatsu, N. Frickhofen, S. Anderson, S. Kajigaya, and NS Young. "First continuous propagation of B19 parvovirus in a cell line." Blood 79, no. 1 (January 1, 1992): 18–24. http://dx.doi.org/10.1182/blood.v79.1.18.bloodjournal79118.

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The pathogenic human parvovirus B19 has extreme tropism for human erythroid progenitor cells and has resisted cultivation in conventional cell lines. We report first propagation of this virus in an erythropoietin-dependent strain of a megakaryoblastic leukemia cell line called UT-7. Virus protein was present in about 5% of cells after 1 week of culture. Appropriate ratios of major and minor capsid proteins were determined by immunoblot, and newly synthesized capsid protein was detected by immunoprecipitation of radioactively labeled cell lysates. High molecular weight monomer and dimer intermediates were detected by Southern analysis, indicating active viral replication. Approximately 1,000 genome copies were present per infected cell, and at the optimal multiplicity of infection 20- to 50- fold more virus was produced than inoculated. Virus propagation only occurred in UT-7 cells that were adapted to growth in erythropoietin; virus signal was not detected in UT-7 cells adapted for growth in granulocyte-macrophage colony-stimulating factor or interleukin-3, even with exposure to erythropoietin for several days. Infectious virus was detected in cultures as long as 3 months after inoculation. Despite persistence, there was no evidence of viral integration on Southern analysis. This cell line may prove useful for the production of infectious virus and in the analysis of B19 parvovirus persistence, cytotoxicity, and permissivity.
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Fabbri, S., F. Marini, C. Mavilia, R. Zonefrati, G. Galli, A. Tanini, M. L. Brandi, and A. R. Gomes. "PTH-C1: A parathyroid continuous cell line derived from rat." Bone 51, no. 6 (December 2012): S28. http://dx.doi.org/10.1016/j.bone.2012.08.102.

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Koyama, Hiroyuki, Yasutaka Imamura, Hiroyasu Yoshikawa, Osamu Kajikawa, Shigeyoshi Itohara, Yoshio Mizuno, Takashi Yoshikawa, and Hiroshi Saito. "Establishment of a Continuous Cell Line Derived from Leukaemic Cattle." Journal of Veterinary Medicine, Series B 33, no. 1-10 (January 12, 1986): 586–96. http://dx.doi.org/10.1111/j.1439-0450.1986.tb00073.x.

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Dissertations / Theses on the topic "Continuous cell line"

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Tric, Mircea [Verfasser], and Stefan [Akademischer Betreuer] Wölfl. "Optical in-line biosensor for long-term continuous glucose monitoring and control in cell culture / Mircea Tric ; Betreuer: Stefan Wölfl." Heidelberg : Universitätsbibliothek Heidelberg, 2017. http://d-nb.info/1177689065/34.

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Tse, Wan-wai. "A study of genomic DNA methylation in immortalized human epithelial cell lines." Click to view the E-thesis via HKUTO, 2008. http://sunzi.lib.hku.hk/hkuto/record/B41290392.

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Tse, Wan-wai, and 謝韻慧. "A study of genomic DNA methylation in immortalized human epithelial cell lines." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2008. http://hub.hku.hk/bib/B41290392.

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Zhang, Hao. "Cytogenetic and molecular alterations in immortalization of normal esophageal epithelial cells." Click to view the E-thesis via HKUTO, 2005. http://sunzi.lib.hku.hk/hkuto/record/B32047010.

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Zhang, Hao, and 張浩. "Cytogenetic and molecular alterations in immortalization of normal esophageal epithelial cells." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B32047010.

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Darbro, Benjamin Will. "Mechanisms of human epithelial cell immortalization and p16NK4a induced telomere independent sencescence." Diss., University of Iowa, 2007. http://ir.uiowa.edu/etd/183.

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Truesdell, Sharon. "Analysis of cell types in continuous cell lines and analysis of the Epidermal Growth Factor Receptor pathway in Drosophila." Connect to resource, 2009. http://hdl.handle.net/1811/37280.

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Fuzzi, Mirko. "Riprogettazione ed ingegnerizzazione di una linea di assemblaggio in ottica Continuous Improvement: il caso Celli UK." Master's thesis, Alma Mater Studiorum - Università di Bologna, 2020.

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Il progetto fondato sul Continuous Improvement, all’interno del quale è stata sviluppata l’esperienza di tirocinio presso l’azienda inglese Celli UK di proprietà del gruppo italiano Celli S.p.a., vede come obiettivo principale quello di implementare ed attuare le tecniche sulle quali si basa la filosofia della Lean Manufacturing, con il fine ultimo di apportare miglioramenti tangibili attinenti al reparto produttivo e misurarne i risultati. L’elaborato della tesi è incentrato sulla riprogettazione ed ingegnerizzazione di una linea di assemblaggio di un particolare modello di spillatrice di birra (DM3), ritenuta critica per l’efficienza globale del reparto produttivo. Inoltre, viene illustrato come vengono messi in pratica strumenti Lean in relazione alla stessa linea target. I risultati ottenuti sono stati misurati tramite determinati KPI relativi alla riduzione del Tempo Ciclo di assemblaggio, all’efficienza della linea ed alla riduzione di pre-assemblaggi. È stato inoltre calcolato il cut-off period dell’investimento che è stato necessario tramite il payback. Oltre a ciò, durante il periodo di tirocinio, è stato implementato un nuovo sistema gestionale per il quale è servito migrare tutti i dati inerenti ai prodotti, le distinte base e le route dal gestionale obsoleto al più recente software ERP Microsoft Dynamics. Successivamente è proseguito il lavoro di creazione degli stessi per i nuovi prodotti acquistati e/o venduti.
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Hyde, Jennie. "The role of prolactin in regulating CCL28 expression /." Diss., CLICK HERE for online access, 2007. http://contentdm.lib.byu.edu/ETD/image/etd1717.pdf.

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Patel, Shakil. "The role of membrane potential dynamics in cell behaviours : investigating the membrane potential dynamics in the Jurkat and HMEC-1 cell lines using the continuous wavelet transform." Thesis, Lancaster University, 2016. http://eprints.lancs.ac.uk/84813/.

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The role of the plasma membrane potential is most commonly associated with the generation of action potentials in excitable cells, however, experimental evidence suggests that this membrane potential is also linked to various behaviours in all cells (Blackiston et al., 2009). These cell behaviours include cell proliferation, cell migration and even cell survival. The membrane potential has been thought to influence these cell behaviours upstream of the classical transduction pathways. Recent evidence suggests that the membrane potential is dynamic rather than static and this dynamic behaviour may encode information on cell behaviours. The whole cell patch clamping technique coupled with the continuous wavelet transform (CWT) technique was used to investigate the presence of fluctuations and oscillations in the membrane potential of Jurkat cells and HMEC-1 cells. The underlying nature of the membrane potential dynamics of Jurkat cells was investigated by perturbing the extracellular concentration of either K+ , Na+ or Cl- . The membrane potential dynamics of proliferating, non-proliferating and activated Jurkat cells was investigated by either varying the culture medium or treating the cells with the concavalin A mitogen. The membrane potential dynamics of HMEC-1 endothelial cells was also investigated. The magnitude of the static membrane potential of proliferating Jurkat cells was significantly more depolarised that non-proliferating Jurkat cells – a trend which has been observed in a wide range of cell types. The membrane potential dynamics appear to be driven by the conductance of ions rather than the magnitude of the static membrane potential per se. In summary, this thesis has proven that the membrane potential varies with cell state and the CWT technique can be used to interrogate recordings of the membrane potential to ascertain information on the membrane potential dynamics that cannot be currently determined by other techniques.
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Books on the topic "Continuous cell line"

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Ian, Freshney R., and Freshney Mary G, eds. Culture of immortalized cells. New York: Wiley-Liss, 1996.

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Tari, Shahryar Rafi. Characterization of AZT transport properties in a continuous renal epithelial cell line. Ottawa: National Library of Canada, 1995.

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Directorate, Canada Drugs. Utilization of continuous cell lines in the manufacture of biologics. Ottawa, Ont: Health and Welfare Canada = Santé et bien-être social Canada, 1990.

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International Association of Biological Standardization. Cell Culture Committee., World Health Organization, and European Society of Animal Cell Technology., eds. Symposium on Continuous Cell Lines as Substrates for Biologicals: Proceedings of a symposium. Basel: Karger, 1989.

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Hayflick, Leonard. Continuous Cell Lines As Substrates for Biologicals (Developments in Biologicals). S Karger Pub, 1989.

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Cox, Josephine H., Stuart Z. Shapiro, Liza Dawson, Cynthia Geppert, Andrew M. Siegel, and M. Patricia D’Souza. Vaccines for The Prevention and Treatment of HIV Infection. Edited by Mary Ann Cohen, Jack M. Gorman, Jeffrey M. Jacobson, Paul Volberding, and Scott Letendre. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780199392742.003.0032.

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While the HIV/AIDS pandemic continues, the overall incidence of HIV infections has fallen through use of antiretroviral therapy (ART) and multiple prevention modalities. To achieve a durable end to the pandemic and avoid the requirement for daily antiretroviral medication over a lifetime, a safe and effective prophylactic vaccine remains essential. This chapter reviews current advances in prophylactic and therapeutic HIV-1 vaccine strategies and the challenges that lie ahead. Recent success in isolation of potent broadly neutralizing antibodies (bnAbs) from infected individuals, the discovery of mechanisms of bnAb induction, and progress in understanding mechanisms of CD8 T-cell killing of HIV-infected cells and the structure of the HIV envelope trimer have opened new strategies for HIV vaccine design. On the therapeutic front, the persistence of HIV reservoirs remains a formidable obstacle to achieving sustained virological remission in HIV-infected individuals after ART is discontinued. Development of a new generation of immune-based therapeutic agents might contribute to a curative intervention. The chapter closes with an overview of ethical challenges in vaccine development and clinical testing.
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(Editor), M. Williams, ed. International Symposium on Continuous Cell Lines - An International Workshop on Current Issues: Proceedings of a Workshop Held at Masur Auditorium N (Developments in Biologicals). S Karger Pub, 1992.

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Canfield, Donald Eugene. What Is It about Planet Earth? Princeton University Press, 2017. http://dx.doi.org/10.23943/princeton/9780691145020.003.0001.

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This chapter begins by highlighting the three basic ingredients for life: energy, the chemical components that make up cells, and water. It then shows that the availability of each of these is linked by special properties of planet Earth. It concludes that Earth is a pretty terrific place for life. It sits comfortably within the habitable zone of the Sun. In addition, its active tectonics both control the temperature of the surface environment, providing a continuous supply of liquid water, and recycle the basic components required to fuel abundant life. The same tectonics may have also provided optimal conditions for the earliest biosphere.
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Wong, Han Hsi, Basma Greef, and Tim Eisen. Treatment of metastatic renal cancer. Edited by James W. F. Catto. Oxford University Press, 2017. http://dx.doi.org/10.1093/med/9780199659579.003.0089.

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Metastatic renal cancer is resistant to standard chemotherapy. Although some patients with indolent disease can be initially managed with observation, the majority of patients will require aggressive treatment soon after diagnosis. Options include cytoreductive nephrectomy, resection of a solitary metastasis in highly selected cases, or systemic therapy options. The TKIs sunitinib and pazopanib are currently the first-line treatments of choice. Whilst axitinib and cabozantinib have important roles in the second line the PD-1 checkpoint inhibitor, nivolumab, is now established as standard second line therapy. Inhibitors of the mammalian target of rapamycin (mTOR) pathway, everolimus and temsirolimus, interleukin-2 as well as the anti-angiogenic antibody bevacizumab have also been shown to be effective. The treatment paradigm of metastatic renal cancer is constantly changing as evidence from clinical trials continues to emerge. With the development of agents addressing novel targets such as T-cell regulation, the future certainly looks brighter for patients diagnosed with this disease.
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Paulson, CAJ. Greenhouse Gas Control Technologies. Edited by RA Durie, DJ Williams, AY Smith, and P. McMullan. CSIRO Publishing, 2001. http://dx.doi.org/10.1071/9780643105027.

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The control of greenhouse gas emissions continues to be a major global problem. It is inter-disciplinary, both in substance and approach, and covers technical, political and economic issues involving governments, industry and the scientific community. These proceedings contain 220 papers presented at the 5th International Conference on Greenhouse Gas Control Technologies (GHGT-5) held in August 2000 at Cairns, Queensland, Australia. The papers cover the capture of carbon dioxide, geological storage of carbon dioxide, ocean storage of carbon dioxide, storage of carbon dioxide with enhanced hydrocarbon recovery, utilisation of carbon dioxide, other greenhouse gases, fuel cells, alternative energy carriers, energy efficiency, life cycle assessments and energy modelling, economics, international and national policy, trading and accounting policy, social and community issues, and reducing emission from industry and power generation.
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Book chapters on the topic "Continuous cell line"

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Gooch, Jan W. "Continuous Cell Line." In Encyclopedic Dictionary of Polymers, 884. New York, NY: Springer New York, 2011. http://dx.doi.org/10.1007/978-1-4419-6247-8_13466.

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Pörtner, Ralf, Armin Bohmann, Ines Lüdemann, and Herbert Märkl. "Metabolic parameters of a hybridoma cell line in batch and continuous cultivation." In Animal Cell Technology: Basic & Applied Aspects, 471–78. Dordrecht: Springer Netherlands, 1993. http://dx.doi.org/10.1007/978-94-011-2044-9_64.

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Victores, S., A. Castillo, E. Faife, Y. Rabasa, Y. Alvarez, L. Rojas, J. Palacio, and A. Figueredo. "Semi-Continuous Cultures as a Tool for Cell Line Characterization During Process Development." In Cells and Culture, 161–64. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-3419-9_26.

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Mukherjee, Tapan Kumar. "Culture of Continuous Cell Lines." In Practical Approach to Mammalian Cell and Organ Culture, 1–61. Singapore: Springer Nature Singapore, 2023. http://dx.doi.org/10.1007/978-981-19-1731-8_11-1.

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Delvigne, Frank, Lucas Henrion, Vincent Vandenbroucke, and Juan Andres Martinez. "Avoiding the All-or-None Response in Gene Expression During E. coli Continuous Cultivation Based on the On-Line Monitoring of Cell Phenotypic Switching Dynamics." In Methods in Molecular Biology, 103–20. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-2930-7_7.

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Jung, Jin G., and Anne Le. "Targeting Metabolic Cross Talk Between Cancer Cells and Cancer-Associated Fibroblasts." In The Heterogeneity of Cancer Metabolism, 205–14. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-65768-0_15.

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AbstractAlthough cancer has classically been regarded as a genetic disease of uncontrolled cell growth, the importance of the tumor microenvironment (TME) [1, 2] is continuously emphasized by the accumulating evidence that cancer growth is not simply dependent on the cancer cells themselves [3, 4] but also dependent on angiogenesis [5–8], inflammation [9, 10], and the supporting roles of cancer-associated fibroblasts (CAFs) [11–13]. After the discovery that CAFs are able to remodel the tumor matrix within the TME and provide the nutrients and chemicals to promote cancer cell growth [14], many studies have aimed to uncover the cross talk between cancer cells and CAFs. Moreover, a new paradigm in cancer metabolism shows how cancer cells act like “metabolic parasites” to take up the high-energy metabolites, such as lactate, ketone bodies, free fatty acids, and glutamine from supporting cells, including CAFs and cancer-associated adipocytes (CAAs) [15, 16]. This chapter provides an overview of the metabolic coupling between CAFs and cancer cells to further define the therapeutic options to disrupt the CAF-cancer cell interactions.
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Buzgariu, Wanda, Jean-Pierre Aubry-Lachainaye, and Brigitte Galliot. "Studying Stem Cell Biology in Intact and Whole-Body Regenerating Hydra by Flow Cytometry." In Methods in Molecular Biology, 373–98. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2172-1_20.

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AbstractThe freshwater Hydra polyp is a versatile model to study whole-body regeneration from a developmental as well as a cellular point of view. The outstanding regenerative capacities of Hydra are based on its three populations of adult stem cells located in the central body column of the animal. There, these three populations, gastrodermal epithelial, epidermal epithelial, and interstitial, continuously cycle in homeostatic conditions, and their activity is locally regulated after mid-gastric bisection. Moreover, they present an unusual cycling behavior with a short G1 phase and a pausing in G2. This particular cell cycle has been studied for a long time with classical microscopic methods. We describe here two flow cytometry methods that provide accurate and reproducible quantitative data to monitor cell cycle regulation in homeostatic and regenerative contexts. We also present a cell sorting procedure based on flow cytometry, whereby stem cells expressing a fluorescent reporter protein in transgenic lines can be enriched for use in applications such as transcriptomic, proteomic, or cell cycle analysis.
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Ui, K., R. Ueda, and T. Miyake. "Continuous Cell Lines from Imaginal Discs of Drosophila Melanogaster." In Invertebrate and Fish Tissue Culture, 251–54. Berlin, Heidelberg: Springer Berlin Heidelberg, 1988. http://dx.doi.org/10.1007/978-3-642-73626-1_59.

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Löwer, Johannes. "Acceptability of continuous cell lines for the production of biologicals." In Animal Cell Technology: Developments Towards the 21st Century, 581–87. Dordrecht: Springer Netherlands, 1995. http://dx.doi.org/10.1007/978-94-011-0437-1_91.

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Leake, Colin J. "Establishing primary cell cultures from disease vectors and maintenance of continuous cell lines." In The Molecular Biology of Insect Disease Vectors, 487–99. Dordrecht: Springer Netherlands, 1997. http://dx.doi.org/10.1007/978-94-009-1535-0_39.

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Conference papers on the topic "Continuous cell line"

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Zhang, Qingwei, Wei Zhang, Donggang Yao, Peter I. Lelkes, and Jack G. Zhou. "The Co-Continuous Micro-Porous PLLA Scaffolds and Their Application for ACL Reconstruction." In ASME 2010 International Mechanical Engineering Congress and Exposition. ASMEDC, 2010. http://dx.doi.org/10.1115/imece2010-38291.

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Anterior cruciate ligament (ACL) reconstructive surgery is a major health concern world-wide because of a large aging population and increased occurrence of sport-related damage. Tissue engineering is a rapidly growing interdisciplinary field that offers a promising new approach for ACL repair. In order to overcome the shortages of current existing surgical fixation devices, we are combining gradient cellular structure (GCS) injection molding technique and biomedical engineering to develop novel surgical fixation devices (screw, anchor, plate, pin, staple, etc.) that not only incorporate bioactive materials such as growth factors, healing drugs and cells, but have natural bone GCS structure, intended to mimic the natural bone and promote bone tissue growth and eventually eliminate the defects associated with existing surgical fixation devices. In this work, a series of novel poly-L-lactic acid (PLLA) scaffolds with micro-porous structure were prepared by injection molding an immiscible polymer blend, with spatially controlled thermal conditioning to adjust the phase size from core to surface. The produced scaffolds were observed under SEM, which shows a co-continuous structure was created successfully through our method. The biocompatibility and the feasibility of produced micro-porous structural PLLA and PLLA/HA scaffolds as a matrix supporting cell growth tested by culturing murine osteoblasts cell line (7F2) for up to 9 days were assessed by Alamar Blue™ assay, which showed that the manufacturing process had no negative effects on cell proliferation. The cell attachment, spreading, migration and proliferation to confluence were assessed by fluorescent nuclear staining with Hoechst 33258. In order to evaluate the functional and cell biological applicability of the micro-porous structural PLLA scaffolds, a subcutaneous biodegradation test was performed through rat model for 1 week and 1 month time period, respectively. Our results showed that the micro-porous structural PLLA scaffolds are non-toxic, and they showed a mild foreign body reaction and complete fibrous encapsulation after implantation. Well created interconnected porous structure and biocompatibility suggest great potential of the micro-porous PLLA scaffolds in application for ACL reconstruction.
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Khan, Mohammed, Xiaolin Chen, and Jie Xu. "Separation of Non-Viable Chinese Hamster Ovary (CHO) Cells Using Dielectrophoresis in a Deterministic Lateral Displacement Ratchet." In ASME 2020 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2020. http://dx.doi.org/10.1115/imece2020-23520.

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Abstract Chinese hamster ovary (CHO) cell is the most widely used mammalian cell line for commercial production of therapeutic protein. Any presence of non-viable cells in culture medium may adversely affect subsequent functionality of these proteins. Therefore, separation of non-viable cells from suspending medium is critical in biopharmaceutical and biomedical sectors. One such method termed Deterministic Lateral Displacement has already shown promising capabilities in separating cells based on the cell size difference by taking advantage of the predictable flow laminae. However, in cases where size overlaps between viable and non-viable cells are present, separation based solely on size suffers and high-resolution separation techniques are required. Dielectrophoresis, one of the most widely used nonlinear electro-kinetic mechanism, has the potential to manipulate the same size cells depending on the dielectric properties of individual cells. In this work, we demonstrated that a DLD device can be combined with a frequency-based AC electric field to perform high resolution continuous separation of non-viable CHO cells from the viable or productive cells. The behavior of the coupled DLD-DEP device is further investigated by employing numerical simulation to check the effect of geometrical parameters of the DLD arrays, velocities of the flow field and required applied voltages. A moderate row shift fraction with velocity 700μm/s provided a good separation behavior without any trapping. The cell viability was also ensured by maintaining proper electric field which otherwise may cause cell loss due to ion leakage. Our developed numerical model and presented results lay the groundwork for design and fabrication of high resolution DLD-DEP microchips for enhanced separation of viable and nonviable cells.
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Morrison, Gerald L., and Mustafa Karabacak. "Evaluation of a Laser Diode for DGV Application." In ASME 2013 Fluids Engineering Division Summer Meeting. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/fedsm2013-16113.

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This paper will present the evaluation of a 300 mw, continuous wave, laser diode system which appears to be suitable for DGV use. The laser frequency range is sufficiently narrow and the lasing frequency can be adjusted across the range of the iodine reference cell (absorption line filter). A spectrum analyzer is used to document the frequency content of the laser across the frequency range of the iodine reference cell. The response of the iodine reference cell to the laser across its spectrum is measured at several iodine reference cell temperatures. The beam profile is also measured. This set of data determined the applicability of this laser for DGV use.
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Sayers, Michael B., and Tara M. Dalton. "A Novel Contamination Free Two Temperature Continuous Flow Polymerase Chain Reactor." In ASME 2007 International Mechanical Engineering Congress and Exposition. ASMEDC, 2007. http://dx.doi.org/10.1115/imece2007-43055.

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Polymerase Chain Reaction (PCR) is an enzymatic process that has dramatically advanced many fields of life sciences, where it is an indispensable tool in a burgeoning range of applications, including diagnostic medicine, molecular biology, forensics and food testing. Recent increased demand for extremely high throughput PCR systems has led to the development of miniaturised continuous flow microfluidic PCR devices, which may have extremely high throughput compared to standard commercial PCR thermal cyclers. A novel continuous flow microfluidic PCR device has been designed and fabricated, consisting of two thermal zones maintained on aluminium thermal blocks providing the precise temperatures required for denaturation and annealing/extension. Polycarbonate sideplates retain the denaturation thermal block vertically above the annealing/extension thermal block while allowing for a variable air gap to be maintained between them. Heating of the denaturation thermal block is achieved using a Labview controlled Thermofoil heater, while the annealing/extension thermal block is maintained at temperature by optimised heat transfer from the denaturation block. Flow-through capillary tubing is positioned into a grooved serpentine channel machined into these thermal blocks. This serpentine channel passes through each thermal block fifty times, providing fifty PCR thermal cycles. Contamination free high throughput continuous flow PCR necessitates that the samples be encapsulated in an immiscible carrier fluid to eradicate cross contamination between samples and suppress the likelihood of the sample contacting the capillary leading to sample degradation. Encapsulation of the PCR reaction mixture is achieved upstream of the thermal cycler through segmentation of the sample into droplets entrained within an immiscible carrier fluid, which are then cycled through the thermal cycler. High throughput DNA amplification of two genes, GAPDH and LEF1, from the REH cell line has been successfully demonstrated on this microfluidic platform without any detectable contamination between samples. The PCR droplet reactors were approximately 250nl which is two orders of magnitude less than the standard sample size for most commercial PCR thermal cyclers.
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Fleming, Paul, and Tara Dalton. "One-Step Reverse-Transcription PCR on a High-Throughput Micro-Fluidic Device." In ASME 2009 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2009. http://dx.doi.org/10.1115/sbc2009-206623.

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One step reverse-transcription polymerase chain reaction (RT-PCR) assays are an attractive option for further automating gene detection assays. One-step assays can reduce hands–on-time and the risk of sample crossover and contamination. The one-step chemistries are showing increasing use in virus detection and have been reported, in some cases, to be more appropriate than their two-step counterparts [1, 2]. Previous work presented by the Stokes Institute research group outlined a micro fluidic based continuous flow instrument which performed high throughput qPCR in nanolitre sized droplets [3]. This instrument had advantages over commercially available instruments in that it could process far more than the traditional 96 or 384 reaction setup in a single run and the reaction volume was reduced from 20–50 μl down to 30–100 nl sized droplets. Combining one-step chemistry with the technology offered by the devices being developed would lead to a high-throughput RNA-to-signal system capable of reverse transcribing and performing PCR on thousands of nanolitre sized reactions every day. It is envisaged that this technology will also lead to gene expression from single cells contained in nanolitre sized droplets. In this paper, a study was conducted in which an extra thermal region, manufactured from aluminium, was added to the existing continuous flow instruments. This region was maintained at a temperature suitable for reverse transcription, which was 48°C for the one-step kit tested. The thermal region was also a suitable length to maintain the sample at the required temperature for 15 minutes. Using a commercially available one step RT-PCR kit (TaqMan® RNA-to-CT™ 1-Step Kit, 4392653), the device was evaluated for its potential to perform one-step RT-PCR in continuously flowing nanolitre sized droplets. Electrophoresis gels were initially used in assessing specific amplification before an end-point detection method was utilized. RNA was extracted from the leukemic REH cell line with the housekeeping gene, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) as the gene of interest. To investigate the possibility of further reducing sample preparation and facilitating further automation, amplification from cell lysates without nucleic acid extraction was carried out on the device. Cell lysates were prepared using the cell lysis buffer from the TaqMan® Gene Expression Cells-to-CT™ Kit (Cat #AM1728). It was found that the device was successful in one-step RT-PCR from extracted RNA samples and samples from cell lysates without nucleic acid extraction.
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Kenner, Scott A., Nicholas M. Josefik, Scott M. Lux, James L. Knight, Melissa K. White, Franklin H. Holcomb, and Gregory J. Ropp. "Component Failure Analysis From the U.S. Army ERDC-CERL Residential Proton Exchange Membrane Fuel Cells Demonstration." In ASME 2006 4th International Conference on Fuel Cell Science, Engineering and Technology. ASMEDC, 2006. http://dx.doi.org/10.1115/fuelcell2006-97245.

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Background: The U.S. Army Engineer Research and Development Center, Construction Engineering Research Laboratory (ERDC-CERL) continues to manage The Department of Defense (DoD) Residential Proton Exchange Membrane (PEM) Fuel Cell Demonstration Project. This project was funded by the United States Congress for fiscal years 2001 through 2004. A fleet of 91 residential-scale PEM fuel cells, ranging in size from 1 to 5 kW, has been demonstrated at various U.S. DoD facilities around the world. Approach: The performance of the fuel cells has been monitored over a 12-month field demonstration period. A detailed analysis has been performed cataloging the component failures, investigating the mean time of the failures, and the mean time between failures. A discussion of the lifespan and failure modes of selected fuel cell components, based on component type, age, and usage will be provided. This analysis also addresses fuel cell stack life for both primary and back-up power systems. Several fuels were used throughout the demonstration, including natural gas, propane, and hydrogen. A distinction will be made on any variances in performance based on the input fuel stock. Summary: This analysis will provide an overview of the ERDC-CERL PEM demonstration fuel cell applications and the corresponding data from the field demonstrations. Special emphasis will be placed on the components, fuel cell stack life, and input fuel characteristics of the systems demonstrated.
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Zhou, Xiaobo, Jun Yang, Meng Wang, and Stephen T. c. Wong. "A novel cell tracking algorithm and continuous hidden Markov model for cell phase identification." In 2006 IEEE/NLM Life Science Systems and Applications Workshop. IEEE, 2006. http://dx.doi.org/10.1109/lssa.2006.250403.

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Hashimoto, Shigehiro, and Takashi Yokomizo. "Tracings of Interaction Between Myoblasts Under Shear Flow in Vitro." In ASME 2021 Fluids Engineering Division Summer Meeting. American Society of Mechanical Engineers, 2021. http://dx.doi.org/10.1115/fedsm2021-65203.

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Abstract How does the group of cells make orientation perpendicular to the flow direction? How does contact with an adjacent cell affect the orientation of the cell? The orientation of a cell according to the neighbor cell under shear flow fields has been traced in vitro. A Couette type flow device with parallel discs was manufactured for the cell culture under the controlled constant wall shear stress. Cells (C2C12: mouse myoblast cell line) were cultured on the lower disc while applying the shear flow in the medium by the upper rotating disc. After culture for 24 hours without flow for adhesion of cells, 2 Pa of the constant wall shear stress was continuously applied in the incubator for 7 days. The behavior of each cell was traced by time-lapse images observed by an inverted phase contrast microscope placed in an incubator. The experiment shows the following results quantitatively by parameters: the contact ratio, and the angle between major axes of cells approximated to ellipsoids. As the ratio of the contact length with the adjacent cell to the pericellular length increases in the two-dimensional projection images, the adjacent cells tend to be oriented in parallel with each other.
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Fowlkes, P. M., P. K. Lund, M. Blake, and J. Snouwaert. "THE REGULATION OF FIBRINOGEN PRODUCTION INVOLVES AT LEAST ONE OTHER HEPATOCYTE GENE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644317.

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It is currently thought that glucocorticosteriods have a direct effect on the transcription of the alpha, beta and gamma fibrinogen genes. However, our studies indicate that while corticosteriods play a role in fibrinogen production, this role is not due to transcriptional activation via glucocorticosteriod receptors. In initial experiments, we compared the levels of fibrinogen mRNA in hepatocytes isolated from hypophysectomized rats to those from control animals. The levels of mRNA in hypophysectomized rats, which produce little ACTH or corticosteriods, were significantly higher than the levels in control animals. Albumin mRNA levels were unaffected by hypophysectomy. These results are in opposition to those which we had anticipated. Based on previously published data, we had thought that physiologic deprivation of corticosteriods would lead to decreased levels of fibrinogen. We propose that these results are related to the negative feedback that corticosteroids have on Hepatocyte Stimulating Factor (HSF) production through a tightly controlled feedback circuit. To investigate the role of corticosteriods in fibrinogen gene regulation, we have conducted experiments with primary hepatocytes in culture and rat FAZA cells (continuous hepatoma cell line). There is a 4 to 5 fold increase in fibrinogen production when these cells are treated with HSF but no change when these cells are treated with dexamethasone alone. However, there is a marked additional increase in the production of fibrinogen with the combination of dexamethasone and HSF. Data gathered through kinetic analysis of this synergistic interaction suggest that the maximum response to HSF requires another gene product whose production is responsive to dexamethasone. Detailed analysis of the rate of transcription of thegamma fibrinogen gene, its processing and mRNA turnover suggests a specific role for this gene product in regulating fibrinogen synthesis. Characterization of this gene product will lead to greater understanding of the regulation of the Acute Phase Reactants.
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Bennedsen, Jacob, and Karen Chang Yan. "On Continuum Based Multiscale Modelling of Engineered Soft Tissue Constructs." In ASME 2018 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2018. http://dx.doi.org/10.1115/imece2018-88482.

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Engineered tissue constructs are assembled through combining scaffolds, cells and biologically active molecules for restoring, maintaining, or improving damaged tissues or whole organs. Cells in engineered tissue constructs often experience mechanical forces during the fabrication process, maturation process, and under in vivo conditions. These mechanical forces/stimuli induce cellular responses and affect cell viability, proliferation, and differentiation. While it is critical to understand the mechanical milieu of cells in tissue constructs, it is also extremely challenging due to the time and length scale span. Multiscale modeling approaches have been emerged to provide linkage among different length scale. One of the approaches is continuum based multiscale modeling to link organ, tissue and cellular levels. A representative volume element (RVE) with periodic or random microstructure serves as a vehicle to connect different length scales. This study focuses on effects of RVE selection, microstructure, and boundary conditions on the mechanical environment at cellular level. In particular, cell embedded alginate tissue constructs were studied. Hyperelastic models were used for modeling alginate and cells. Multi-cellular FE models were generated. The results of the average properties and the stress/strain experienced by cells were compared under different conditions.
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Reports on the topic "Continuous cell line"

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Petitte, James, Hefzibah Eyal-Giladi, and Malka Ginsburg. The Study of Primordial Germ Cell Development as a Tool for Gene Transfer in Chickens. United States Department of Agriculture, October 1991. http://dx.doi.org/10.32747/1991.7561071.bard.

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The ability to introduce novel genetic material into the genome of commercial poultry has been impeded by a lack of kowledge regarding the origin in the early embryo of the target cell of interest, namely, the germ cell. Hence, this project investigated the emergence of primordial germ cells (PGCs) during the early development of the avian embryo to aid in efforts to produce transgenic poultry on a routine basis. The strategy was to introduce foreign DNA into the area of the unincubated embryo that is destined to give rise to the germ line. The objectives of this project were: 1) to identify and localize a subpopulation of cells in the early embryo which will give rise to PGCs, 2) to determine the best location and stage of development to transfer donor cells for efficient germline chimerism, and 3) to transfect donor cells to produce transgenic/germline chimeric embryos. We show that by using the monoclonal antibody SSEA-1 and by various cell culture techniques that germ cells appear to segregate from the somatic lineages at St. X., a process that is gradual and continues through St. XIV. Using microsurgical transplantation between quail and chick embryos, we demonstrated that the inner 1/3 of the area pellucida between states X-XII gives rise to about 2/3 of the germ cell population at the time of their residence in the germinal crescent. Because of the non-localized emergence of PGCs, attempts to introduce foreign DNA into clonal precursors of germ cells through liposome-mediated transfection yielded unacceptable levels of efficiency. However, through our investigation of germ cell origins, an in vitro model of germ cell differentiation was developed that could offer a means of determining the factors required for the long term culture of avian PGCs thereby providing a convenient means of manipulating the avian genome.
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Eshed-Williams, Leor, and Daniel Zilberman. Genetic and cellular networks regulating cell fate at the shoot apical meristem. United States Department of Agriculture, January 2014. http://dx.doi.org/10.32747/2014.7699862.bard.

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The shoot apical meristem establishes plant architecture by continuously producing new lateral organs such as leaves, axillary meristems and flowers throughout the plant life cycle. This unique capacity is achieved by a group of self-renewing pluripotent stem cells that give rise to founder cells, which can differentiate into multiple cell and tissue types in response to environmental and developmental cues. Cell fate specification at the shoot apical meristem is programmed primarily by transcription factors acting in a complex gene regulatory network. In this project we proposed to provide significant understanding of meristem maintenance and cell fate specification by studying four transcription factors acting at the meristem. Our original aim was to identify the direct target genes of WUS, STM, KNAT6 and CNA transcription factor in a genome wide scale and the manner by which they regulate their targets. Our goal was to integrate this data into a regulatory model of cell fate specification in the SAM and to identify key genes within the model for further study. We have generated transgenic plants carrying the four TF with two different tags and preformed chromatin Immunoprecipitation (ChIP) assay to identify the TF direct target genes. Due to unforeseen obstacles we have been delayed in achieving this aim but hope to accomplish it soon. Using the GR inducible system, genetic approach and transcriptome analysis [mRNA-seq] we provided a new look at meristem activity and its regulation of morphogenesis and phyllotaxy and propose a coherent framework for the role of many factors acting in meristem development and maintenance. We provided evidence for 3 different mechanisms for the regulation of WUS expression, DNA methylation, a second receptor pathway - the ERECTA receptor and the CNA TF that negatively regulates WUS expression in its own domain, the Organizing Center. We found that once the WUS expression level surpasses a certain threshold it alters cell identity at the periphery of the inflorescence meristem from floral meristem to carpel fate [FM]. When WUS expression highly elevated in the FM, the meristem turn into indeterminate. We showed that WUS activate cytokinine, inhibit auxin response and represses the genes required for root identity fate and that gradual increase in WUCHEL activity leads to gradual meristem enlargement that affect phyllotaxis. We also propose a model in which the direction of WUS domain expansion laterally or upward affects meristem structure differently. We preformed mRNA-seq on meristems with different size and structure followed by k-means clustering and identified groups of genes that are expressed in specific domains at the meristem. We will integrate this data with the ChIP-seq of the 4 TF to add another layer to the genetic network regulating meristem activity.
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James, Christian, Ronald Dixon, Luke Talbot, Stephen James, Nicola Williams, and Bukola Onarinde. Assessing the impact of heat treatment on antimicrobial resistant (AMR) genes and their potential uptake by other ‘live’ bacteria. Food Standards Agency, August 2021. http://dx.doi.org/10.46756/sci.fsa.oxk434.

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Addressing the public health threat posed by AMR is a national strategic priority for the UK, which has led to both a 20-year vision of AMR and a 5-year (2019 to 2024) AMR National Action Plan (NAP). The latter sets out actions to slow the development and spread of AMR with a focus on antimicrobials. The NAP used an integrated ‘One-Health’ approach which spanned people, animals, agriculture and the environment, and calls for activities to “identify and assess the sources, pathways, and exposure risks” of AMR. The FSA continues to contribute to delivery of the NAP in a number of ways, including through furthering our understanding of the role of the food chain and AMR.Thorough cooking of food kills vegetative bacterial cells including pathogens and is therefore a crucial step in reducing the risk of most forms of food poisoning. Currently, there is uncertainty around whether cooking food is sufficient to denature AMR genes and mobile genetic elements from these ‘dead’ bacteria to prevent uptake by ‘live’ bacteria in the human gut and other food environments - therefore potentially contributing to the overall transmission of AMR to humans. This work was carried out to assess these evidence gaps.
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Fahima, Tzion, and Jorge Dubcovsky. Map-based cloning of the novel stripe rust resistance gene YrG303 and its use to engineer 1B chromosome with multiple beneficial traits. United States Department of Agriculture, January 2013. http://dx.doi.org/10.32747/2013.7598147.bard.

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Research problem: Bread wheat (Triticumaestivum) provides approximately 20% of the calories and proteins consumed by humankind. As the world population continues to increase, it is necessary to improve wheat yields, increase grain quality, and minimize the losses produced by biotic and abiotic stresses. Stripe rust, caused by Pucciniastriiformisf. sp. tritici(Pst), is one of the most destructive diseases of wheat. The new pathogen races are more virulent and aggressive than previous ones and have produced large economic losses. A rich source for stripe-rust resistance genes (Yr) was found in wild emmer wheat populations from Israel. Original Project goals: Our long term goal is to identify, map, clone, characterize and deploy in breeding, novel wild emmer Yr genes, and combine them with multiple beneficial traits. The current study was aiming to map and clone YrG303 and Yr15, located on chromosome 1BS and combine them with drought resistance and grain quality genes. Positional cloning of YrG303/Yr15: Fine mapping of these genes revealed that YrG303 is actually allelic to Yr15. Fine genetic mapping using large segregating populations resulted in reduction of the genetic interval spanning Yr15 to less than 0.1 cM. Physical mapping of the YrG303/Yr15 locus was based on the complete chromosome 1BS physical map of wheat constructed by our group. Screening of 1BS BAC library with Yr15 markers revealed a long BAC scaffold covering the target region. The screening of T. dicoccoidesaccession-specific BAC library with Yr15 markers resulted in direct landing on the target site. Sequencing of T. dicoccoidesBAC clones that cover the YrG303/Yr15 locus revealed a single candidate gene (CG) with conserved domains that may indicate a role in disease resistance response. Validation of the CG was carried out using EMS mutagenesis (loss-of- function approach). Sequencing of the CG in susceptible yr15/yrG303 plants revealed three independent mutants that harbour non-functional yr15/yrG303 alleles within the CG conserved domains, and therefore validated its function as a Pstresistance gene. Evaluation of marker-assisted-selection (MAS) for Yr15. Introgressions of Yr15 into cultivated wheat are widely used now. Recently, we have shown that DNA markers linked to Yr15 can be used as efficient tools for introgression of Yr15 into cultivated wheat via MAS. The developed markers were consistent and polymorphic in all 34 tested introgressions and are the most recommended markers for the introgression of Yr15. These markers will facilitate simultaneous selection for multiple Yr genes and help to avoid escapees during the selection process. Engineering of improved chromosome 1BS that harbors multiple beneficial traits. We have implemented the knowledge and genetic resources accumulated in this project for the engineering of 1B "super-chromosome" that harbors multiple beneficial traits. We completed the generation of a chromosome including the rye 1RS distal segment associated with improved drought tolerance with the Yr gene, Yr15, and the strong gluten allele 7Bx-over-expressor (7Bxᴼᴱ). We have completed the introgression of this improved chromosome into our recently released variety Patwin-515HP and our rain fed variety Kern, as well as to our top breeding lines UC1767 and UC1745. Elucidating the mechanism of resistance exhibited by Yr36 (WKS1). The WHEAT KINASE START1 (WKS1) resistance gene (Yr36) confers partial resistance to Pst. We have shown that wheat plants transformed with WKS1 transcript are resistant to Pst. WKS1 is targeted to the chloroplast where it phosphorylates the thylakoid-associatedascorbateperoxidase (tAPX) and reduces its ability to detoxify peroxides. Based on these results, we propose that the phosphorylation of tAPX by WKS1 reduces the ability of the cells to detoxify ROS and contributes to cell death. Distribution and diversity of WKS in wild emmer populations. We have shown that WKS1 is present only in the southern distribution range of wild emmer in the Fertile Crescent. Sequence analysis revealed a high level of WKS1 conservation among wild emmer populations, in contrast to the high level of diversity observed in NB-LRR genes. This phenomenon shed some light on the evolution of genes that confer partial resistance to Pst. Three new WKS1 haplotypes displayed a resistance response, suggesting that they can be useful to improve wheat resistance to Pst. In summary, we have improved our understanding of cereals’ resistance mechanisms to rusts and we have used that knowledge to develop improved wheat varieties.
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Splitter, Gary, and Menachem Banai. Microarray Analysis of Brucella melitensis Pathogenesis. United States Department of Agriculture, 2006. http://dx.doi.org/10.32747/2006.7709884.bard.

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Original Objectives 1. To determine the Brucella genes that lead to chronic macrophage infection. 2. To identify Brucella genes that contribute to infection. 3. To confirm the importance of Brucella genes in macrophages and placental cells by mutational analysis. Background Brucella spp. is a Gram-negative facultative intracellular bacterium that infects ruminants causing abortion or birth of severely debilitated animals. Brucellosis continues in Israel, caused by B. melitensis despite an intensive eradication campaign. Problems with the Rev1 vaccine emphasize the need for a greater understanding of Brucella pathogenesis that could improve vaccine designs. Virulent Brucella has developed a successful strategy for survival in its host and transmission to other hosts. To invade the host, virulent Brucella establishes an intracellular niche within macrophages avoiding macrophage killing, ensuring its long-term survival. Then, to exit the host, Brucella uses placenta where it replicates to high numbers resulting in abortion. Also, Brucella traffics to the mammary gland where it is secreted in milk. Missing from our understanding of brucellosis is the surprisingly lillie basic information detailing the mechanisms that permit bacterial persistence in infected macrophages (chronic infection) and dissemination to other animals from infected placental cells and milk (acute infection). Microarray analysis is a powerful approach to determine global gene expression in bacteria. The close genomic similarities of Brucella species and our recent comparative genomic studies of Brucella species using our B. melitensis microarray, suqqests that the data obtained from studying B. melitensis 16M would enable understanding the pathogenicity of other Brucella organisms, particularly the diverse B. melitensis variants that confound Brucella eradication in Israel. Conclusions Results from our BARD studies have identified previously unknown mechanisms of Brucella melitensis pathogenesis- i.e., response to blue light, quorum sensing, second messenger signaling by cyclic di-GMP, the importance of genomic island 2 for lipopolysaccharide in the outer bacterial membrane, and the role of a TIR domain containing protein that mimics a host intracellular signaling molecule. Each one of these pathogenic mechanisms offers major steps in our understanding of Brucella pathogenesis. Strikingly, our molecular results have correlated well to the pathognomonic profile of the disease. We have shown that infected cattle do not elicit antibodies to the organisms at the onset of infection, in correlation to the stealth pathogenesis shown by a molecular approach. Moreover, our field studies have shown that Brucella exploit this time frame to transmit in nature by synchronizing their life cycle to the gestation cycle of their host succumbing to abortion in the last trimester of pregnancy that spreads massive numbers of organisms in the environment. Knowing the bacterial mechanisms that contribute to the virulence of Brucella in its host has initiated the agricultural opportunities for developing new vaccines and diagnostic assays as well as improving control and eradication campaigns based on herd management and linking diagnosis to the pregnancy status of the animals. Scientific and Agricultural Implications Our BARD funded studies have revealed important Brucella virulence mechanisms of pathogenesis. Our publication in Science has identified a highly novel concept where Brucella utilizes blue light to increase its virulence similar to some plant bacterial pathogens. Further, our studies have revealed bacterial second messengers that regulate virulence, quorum sensing mechanisms permitting bacteria to evaluate their environment, and a genomic island that controls synthesis of its lipopolysaccharide surface. Discussions are ongoing with a vaccine company for application of this genomic island knowledge in a Brucella vaccine by the U.S. lab. Also, our new technology of bioengineering bioluminescent Brucella has resulted in a spin-off application for diagnosis of Brucella infected animals by the Israeli lab by prioritizing bacterial diagnosis over serological diagnosis.
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Hochman, Ayala, Thomas Nash III, and Pamela Padgett. Physiological and Biochemical Characterization of the Effects of Oxidant Air Pollutants, Ozone and Gas-phase Nitric Acid, on Plants and Lichens for their Use as Early Warning Biomonitors of these Air Pollutants. United States Department of Agriculture, January 2011. http://dx.doi.org/10.32747/2011.7697115.bard.

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Introduction. Ozone and related oxidants are regarded as the most important phytotoxic air pollutant in many parts of the western world. A previously unrecognized component of smog, nitric acid, may have even greater deleterious effects on plants either by itself or by augmenting ozone injury. The effects of ozone on plants are well characterized with respect to structural and physiological changes, but very little is known about the biochemical changes in plants and lichens exposed to ozone and/or HNO3. Objectives.To compare and contrast the responses of crop plants and lichens to dry deposition of HNO3 and O3., separately, and combined in order to assess our working hypothesis that lichens respond to air pollution faster than plants. Lichens are most suitable for use as biomonitors because they offer a live-organism-based system that does not require maintenance and can be attached to any site, without the need for man-made technical support systems. Original Immediate aims To expose the tobacco (Nicotiana tabacum L.) cultivar Bel-W3 that is ozone supersensitive and the ozone sensitive red kidney bean (Phaseolusvulgaris) and the lichen Ramalinamenziesii to controlled HNO3 and O3 fumigations and combined and to follow the resulting structural, physiological and biochemical changes, with special reference to reactive oxygen species related parameters. Revised. Due to technical problems and time limitations we studied the lichen Ramalinamenziesii and two cultivar of tobacco: Bel-W3 that is ozone supersensitive and a resistant cultivar, which were exposed to HNO3 and O3 alone (not combined). Methodology. Plants and lichens were exposed in fumigation experiments to HNO3 and O3, in constantly stirred tank reactors and the resulting structural, physiological and biochemical changes were analyzed. Results. Lichens. Exposure of Ramalinamenziesiito HNO3 resulted in cell membrane damage that was evident by 14 days and continues to worsen by 28 days. Chlorophyll, photosynthesis and respiration all declined significantly in HNO3 treatments, with the toxic effects increasing with dosage. In contrast, O3 fumigations of R. menziesii showed no significant negative effects with no differences in the above response variables between high, moderate and low levels of fumigations. There was a gradual decrease in catalase activity with increased levels of HNO3. The activity of glutathione reductase dropped to 20% in thalli exposed to low HNO3 but increased with its increase. Glucose 6-phosphate dehydrogenase activity increase by 20% with low levels of the pollutants but decreased with its increase. Tobacco. After 3 weeks of exposure of the sensitive tobacco cultivar to ozone there were visible symptoms of toxicity, but no danmage was evident in the tolerant cultivar. Neither cultivar showed any visible symptoms after exposure to HNO3.In tobacco fumigated with O3, there was a significant decrease in maximum photosynthetic CO2 assimilation and stomatal conductance at high levels of the pollutant, while changes in mesophyll conductance were not significant. However, under HNO3 fumigation there was a significant increase in mesophyll conductance at low and high HNO3 levels while changes in maximum photosynthetic CO2 assimilation and stomatal conductance were not significant. We could not detect any activity of the antioxidant enzymes in the fumigated tobacco leaves. This is in spite of the fact that we were able to assay the enzymes in tobacco leaves grown in Israel. Conclusions. This project generated novel data, and potentially applicable to agriculture, on the differential response of lichens and tobacco to HNO3 and O3 pollutants. However, due to experimental problems and time limitation discussed in the body of the report, our data do not justify yet application for a full, 4-year grant. We hope that in the future we shall conduct more experiments related to our objectives, which will serve as a basis for a larger scale project to explore the possibility of using lichens and/or plants for biomonitoring of ozone and nitric acid air pollution.
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