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Letteratura scientifica selezionata sul tema "Stem cells – Research"

Autore: Grafiati

Pubblicato: 4 giugno 2021

Ultima modifica: 31 gennaio 2023

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Articoli di riviste sul tema "Stem cells – Research"

Rahat, Hashmi, e Ahmad Fahim. "Stem Cells: A Gold Mine in Dental Research and Tissue Engineering". Cancer Medicine Journal 2, n. 2 (31 dicembre 2019): 41–44. http://dx.doi.org/10.46619/cmj.2019.2-1012.

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The explosion of articles not only in scientific journals, but also in the print media and continuous TV debates, one could easily say the term “stem cells” has become synonym to the word “cure”. The extraordinary advances in the prevention, diagnosis, and treatment of human diseases, such as oral health issues, cancer, heart disease, diabetes and diseases of the central and peripheral nervous system, such as Parkinson's disease and Alzheimer's disease, continues to deprive people of health and well-being. Tremendous effort and exceptional research in human developmental biology has led to the identification and discovery of human stem cells.

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Stojkovic, Miodrag, Mark F. Pittenger, Jan A. Nolta, Majlinda Lako, T. R. J. Lappin e Martin J. Murphy. "STEM CELLS' Position Statement on hESC Research". STEM CELLS 28, n. 9 (settembre 2010): 1A. http://dx.doi.org/10.1002/stem.517.

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Caulfield, Timothy, Kalina Kamenova, Ubaka Ogbogu, Amy Zarzeczny, Jay Baltz, Shelly Benjaminy, Paul A. Cassar et al. "Research ethics and stem cells". EMBO reports 16, n. 1 (4 dicembre 2014): 2–6. http://dx.doi.org/10.15252/embr.201439819.

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Ikram, Huma, Darakhshan Jabeen Haleem, Zia Choudhry e Adnan Maqsood Choudhry. "Stem cells in Parkinson's research". El Mednifico Journal 1, n. 3 (7 ottobre 2013): 77. http://dx.doi.org/10.18035/emj.v1i3.39.

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King, Anthony. "Stem cells: highlights from research". Nature 597, n. 7878 (29 settembre 2021): S6—S7. http://dx.doi.org/10.1038/d41586-021-02621-4.

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Bolt, Hermann M. "Stem cells in toxicological research". Archives of Toxicology 91, n. 12 (15 novembre 2017): 4029–30. http://dx.doi.org/10.1007/s00204-017-2120-9.

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Grisham, Julie. "Stem cells as research tools". Nature Biotechnology 18, n. 4 (aprile 2000): 366. http://dx.doi.org/10.1038/74350.

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Holden, C. "STEM CELLS: U.S. Public Supports Stem Cell Research". Science 310, n. 5747 (21 ottobre 2005): 416b. http://dx.doi.org/10.1126/science.310.5747.416b.

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Holden, C. "STEM CELL RESEARCH: Primate Parthenotes Yield Stem Cells". Science 295, n. 5556 (1 febbraio 2002): 779a—780. http://dx.doi.org/10.1126/science.295.5556.779a.

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Revel, Michel. "Research on Human embryonic stem cells and cloning for stem cells". Human Reproduction & Genetic Ethics 14, n. 1 (29 luglio 2008): 4–14. http://dx.doi.org/10.1558/hrge.v14i1.4.

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Tesi sul tema "Stem cells – Research"

Marshall, Gregory Paul. "Neurospheres and multipotent astrocytic stem cells neural progenitor cells rather than neural stem cells /". [Gainesville, Fla.] : University of Florida, 2005. http://purl.fcla.edu/fcla/etd/UFE0010047.

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Thesis (Ph.D.)--University of Florida, 2005.
Typescript. Title from title page of source document. Document formatted into pages; contains 97 pages. Includes Vita. Includes bibliographical references.

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Elichabe, Benoît. "United States stem cells research boundaries". Thesis, Massachusetts Institute of Technology, 2007. http://hdl.handle.net/1721.1/39529.

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Thesis (M.B.A.)--Massachusetts Institute of Technology, Sloan School of Management, 2007.
Includes bibliographical references (leaves [88]-90).
Recent empirical work has demonstrated the importance of a number of elements of scientific infrastructure that seem to be crucial particularly in fields such as molecular and cellular biology in which the materiality of research renders the process of replication and validation more complex. Scientific infrastructure has many interconnecting elements such as the ability to exchange material used in experiments, the ability to share ideas and information and the ability to share, exchange and promote the mobility of researchers. We focus our investigation on stem cell research in the United States (US). Research in human developmental biology has led to the discovery of human stem cells. The science of stem cell therapies is about to enter a phase of research and development that could lead to unprecedented cures and palliative treatments. However, it is a highly regulated field of research and it raises an important amount of moral, religious and ethical concerns. We seek to examine the boundaries that have emerged in the US in this particular field and we try to understand their impact on the US market of fertilized eggs, embryos and human embryonic stem cells.
by Benoît Elichabe.
M.B.A.

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Nortjé, Nico. "The moral status of embryonic stem cell research in the South African context /". Link to the online version, 2007. http://hdl.handle.net/10019.1/1372.

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Chen, Shi. "Cryopreservation of human embryonic stem cells and hepatocytes". Thesis, University of Oxford, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.711665.

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Ngangan, Alyssa V. "Bioactive factors secreted by differentiating embryonic stem cells". Diss., Georgia Institute of Technology, 2011. http://hdl.handle.net/1853/44913.

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Current therapeutic strategies to stimulate endogenous angiogenic processes within injured tissue areas are typically based on introducing exogenous pro-angiogenic molecules or cell populations. Stem cell transplantation for angiogenic therapy aims to deliver populations of cells that secrete angiogenic factors and/or engraft in the new branching vasculature within the damaged tissue. Utilizing stem or progenitor cells has been shown to induce a rather robust angiogenic response despite minimal repopulation of the host vasculature, suggesting that stem cells may provide paracrine factors that transiently induce endogenous angiogenesis of tissues undergoing regeneration. Early differentiating embryonic stem cell (ESC) aggregates, referred to as embryoid bodies (EBs), can undergo vasculogenic differentiation, and also produce extracellular matrix and growth factors that induce proliferation, differentiation, and tissue morphogenesis. Taken together, the ESC extracellular environment may be an effective means by which to manipulate cell behavior. Thus, the objective of this project was to harness morphogens derived from ESCs undergoing differentiation and analyze their bioactive potential. To examine the expression of extracellular factors within EBs, gene expression arrays in conjunction with a variety of analytical tools were utilized to gain an understanding of the importance of extracellular factors in ESC differentiation. Furthermore, the soluble fraction of secreted factors contained within EB-conditioned media was compared to the matrix-associated factors produced by EBs, which led to the development of novel ESC-derived matrices via mechanical acellularization methods. Acellular embryonic stem cell-derived matrices demonstrated the retention of bioactive factors that impacted aspects of angiogenesis. In conclusion, extracellular factors were modulated in response to the progression of EB differentiation and can further be harnessed via acellularization techniques, in order to deliver bioactive ESC-secreted factors in a cell-free manner.

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Ericson, Robin J. "Bridging solutions to the religion and science conflict over human embryonic stem cell research". Fairfax, VA : George Mason University, 2007. http://hdl.handle.net/1920/2926.

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Thesis (Ph. D.)--George Mason University, 2007.
Title from PDF t.p. (viewed Jan. 17, 2008). Thesis director: Richard E. Rubenstein. Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Conflict Analysis and Resolution. Vita: p. 228. Includes bibliographical references (p. 222-227). Also available in print.

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Alawadhi, Aseel. "Human embryonic stem cell research : shaping regulations in Kuwait". Thesis, University of Aberdeen, 2016. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=231070.

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Ke, Jin, e 柯金. "Transgenic stem cells for craniofacial bone reconstruction". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2010. http://hub.hku.hk/bib/B44362973.

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Bone loss from the cranio-maxillofacial region is a major clinical problem affecting patients worldwide. Conventional treatment strategy includes the use of autogenous or allogeneic bone, biomaterials, and osteogenic growth factors. However, there has been no effective therapy for most cases so far. Stem cell-based gene therapy is the latest research method with possible applications in humans. The present study aims to (1) characterize rabbit mesenchymal stem cells (MSCs) relating to growth pattern, surface antigens, and the potential for multi-differentiation; (2) determine the transduction efficiency and duration of recombinant adeno-associated virus2 carrying enhanced green fluorescent protein (rAAV2EGFP) reporter gene in rabbit MSCs and study the effects of rAAV2EGFP transduction on stem cells’ phenotype and capacity of multi-differentiation; (3) evaluate the differentiation characteristics of rabbit MSCs following recombinant adeno-associated virus 2 carrying bone morphogenetic protein 2 gene (rAAV2BMP2) transduction; (4) investigate whether MSCs transduced by rAAV2BMP2 could successfully induce bone regeneration in rabbit critical-size cranial defects. MSCs were isolated from bone marrows of rabbit tibias and cultured. Cell counting and colony-forming assays demonstrated that growth rates of MSCs dropped substantially with increasing passages. Flow cytometry on MSCs at passage 1 showed that cells expressed high level of CD49a and low level of CD44 as well as stage-specific embryonic antigen 4 (SSEA4). Multi-differentiation and reverse transcriptase-polymerase chain reaction (RTPCR) tests demonstrated that rabbit MSCs were capable to differentiate into osteocytes, chondrocytes and adipocytes. Immunofluorescence microscopy showed that rabbit MSCs produced a series of hematopoietic growth factors, including stem cell factor (SCF), vascular endothelial growth factor-A (VEGFA) and granulocyte macrophage colony-stimulating factor (GMCSF). Subsequently, rabbit MSCs were transduced with rAAV2EGFP in vitro. By comparing the transduction efficiency with different doses of rAAV2EGFP particles, multiplicity of infection (MOI) of 1 x 10 4 was identified as an optimal parameter for the transduction of rAAV2 in rabbit MSCs. Fluorescent microscopy demonstrated long-term expression of EGFP in rabbit MSCs after transduction both in vitro and in vivo. In addition, cell proliferation assay, adipogenic induction test and flow cytometry showed that rAAV2EGFP transduced MSCs exhibited a similar pattern with non-transduced cells on the cell growth, capacity of adipogenic differentiation and expression of surface antigens, indicating that rabbit MSCs maintain their stem cell properties after rAAV2EGFP transduction.
published_or_final_version
Dentistry
Doctoral
Doctor of Philosophy

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Renszel, Krystal Marie. "USING MUTAGENESIS AND STEM CELLS TO UNDERSTAND RETROVIRAL NEUROVIRULENCE". Kent State University / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=kent1254659655.

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Sukhdeo, Kumar. "Defining immunophenotypic signatures of stem cells". Case Western Reserve University School of Graduate Studies / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=case1373038255.

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Più fonti

Libri sul tema "Stem cells – Research"

V, Greer Erik, a cura di. Focus on stem cell research. Hauppauge, N.Y: Nova Biomedical Books, 2004.

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S, Koka Prasad, a cura di. Developments in stem cell research. New York: Nova Science Publishers, 2008.

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V, Greer Erik, a cura di. Embryonic stem cell research. New York: Nova Science Publishers, 2006.

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S, Koka Prasad, a cura di. Progress in stem cell research. New York: Nova Science Publishers, 2008.

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Jacqueline, Langwith, a cura di. Stem cells. Detroit: Greenhaven Press, 2007.

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Stem cells. Detroit: Greenhaven Press, 2012.

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Marcovitz, Hal. Stem cell research. San Diego, CA: ReferencePoint Press, 2011.

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Marcovitz, Hal. Stem cell research. San Diego, CA: ReferencePoint Press, 2011.

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Stem cell research. New York: Rosen, 2003.

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Stem cell research. Detroit: Greenhaven Press, 2012.

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Capitoli di libri sul tema "Stem cells – Research"

Tu, Shi-Ming. "Stem Cells". In Cancer Treatment and Research, 43–54. Boston, MA: Springer US, 2010. http://dx.doi.org/10.1007/978-1-4419-5968-3_5.

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Chowdhury, Suchandra, e Shyamasree Ghosh. "Stem Cells in Clinical Research and Therapy". In Stem Cells, 239–52. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-16-1638-9_10.

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Uçkan-Çetinkaya, Duygu, e Khawaja Husnain Haider. "Induced Pluripotent Stem Cells in Pediatric Research and Clinical Translation". In Stem Cells, 203–16. Cham: Springer International Publishing, 2021. http://dx.doi.org/10.1007/978-3-030-77052-5_13.

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Tu, Shi-Ming. "Cancer Stem Cells". In Cancer Treatment and Research, 67–81. Boston, MA: Springer US, 2010. http://dx.doi.org/10.1007/978-1-4419-5968-3_7.

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Moore, Jennifer C., Teun P. de Boer, Marcel A. G. van der Heyden, Leon G. J. Tertoolen e Christine L. Mummery. "Stem Cells and Cardiomyocytes". In Cardiovascular Research, 133–55. Boston, MA: Springer US, 2006. http://dx.doi.org/10.1007/0-387-23329-6_8.

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Azizi, Hossein, Sabine Conrad, Thomas Skutella e Irma Virant-Klun. "Spermatogonial Stem Cells". In Advances in Stem Cell Research, 191–210. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-940-2_11.

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Koch, Valerie Gutmann, Beth E. Roxland, Barbara Pohl e Sarah K. Keech. "Contemporary Ethical Issues in Stem Cell Research". In Stem Cells Handbook, 29–37. New York, NY: Springer New York, 2013. http://dx.doi.org/10.1007/978-1-4614-7696-2_2.

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Xie, Xiaoyan, Yanhua Li, Yanxun Sun, Jin Zhang, Fang Fang, Wen Yue e Xuetao Pei. "Stem Cells and Hematopoietic Cell Engineering". In Translational Medicine Research, 111–44. Dordrecht: Springer Netherlands, 2015. http://dx.doi.org/10.1007/978-94-017-7273-0_5.

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Da Silva, Jose S., e Joshua M. Hare. "Mesenchymal Stem Cells". In Manual of Research Techniques in Cardiovascular Medicine, 104–9. Oxford, UK: John Wiley & Sons, Ltd, 2013. http://dx.doi.org/10.1002/9781118495148.ch11.

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Löser, Peter, Anke Guhr, Sabine Kobold, Anna M. Wobus e Andreas Kurtz. "Status and Impact of Research on Human Pluripotent Stem Cells: Cell Lines and Their Use in Published Research". In Stem Cells and Cancer Stem Cells, Volume 2, 61–74. Dordrecht: Springer Netherlands, 2011. http://dx.doi.org/10.1007/978-94-007-2016-9_7.

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Atti di convegni sul tema "Stem cells – Research"

Billing, Anja M., Shaima S. Dib, Hisham Ben Hamidane, Neha Goswami, Rasha Al-mismar, Richard Cotton, Pankaj Kumar et al. "Comprehensive Characterization Of The Differentiation Of Human Embryonic Stem Cells Into Mesenchymal Stem Cells". In Qatar Foundation Annual Research Conference Proceedings. Hamad bin Khalifa University Press (HBKU Press), 2014. http://dx.doi.org/10.5339/qfarc.2014.hbpp0979.

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Crous, Anine, e Heidi Abrahamse. "Photodynamic Therapy and Lung Cancer Stem Cells – The effects of AlPcS4Cl on Isolated Lung Cancer Stem Cells". In International e-Conference on Cancer Research 2019. European High-tech and Emerging Research Association, 2019. http://dx.doi.org/10.28991/iccr-2019-012.

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Singh, Ankur, Shalu Suri, Ted T. Lee, Jamie M. Chilton, Steve L. Stice, Hang Lu, Todd C. McDevitt e Andrés J. Garcia. "Adhesive Signature-Based, Label-Free Isolation of Human Pluripotent Stem Cells". In ASME 2012 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2012. http://dx.doi.org/10.1115/sbc2012-80044.

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Generation of human induced pluripotent stem cells (hiPSCs) from fibroblasts and other somatic cells represents a highly promising strategy to produce auto- and allo-genic cell sources for therapeutic approaches as well as novel models of human development and disease1. Reprogramming protocols involve transduction of the Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc into the parental somatic cells, followed by culturing the transduced cells on mouse embryonic fibroblast (MEF) or human fibroblast feeder layers, and subsequent mechanical dissociation of pluripotent cell-like colonies for propagation on feeder layers1, 2. The presence of residual parental and feeder-layer cells introduces experimental variability, pathogenic contamination, and promotes immunogenicity3. Similar to human embryonic stem cells (hESCs), reprogrammed hiPSCs suffer from the unavoidable problem of spontaneous differentiation due to sub-optimal feeder cultures4, growth factors5, and the feeder-free substrate6. Spontaneously differentiated (SD)-hiPSCs display reduced pluripotency and often contaminate hiPSC cultures, resulting in overgrowth of cultures and compromising the quality of residual pluripotent stem cells5. Therefore, the ability to rapidly and efficiently isolate undifferentiated hiPSCs from the parental somatic cells, feeder-layer cells, and spontaneously differentiated cells is a crucial step that remains a bottleneck in all human pluripotent stem cell research.

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Sottoriva, Andrea, e Simon Tavaré. "Abstract PL04-03: Cancer stem cells". In Abstracts: AACR International Conference on Frontiers in Cancer Prevention Research‐‐ Nov 7-10, 2010; Philadelphia, PA. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1940-6207.prev-10-pl04-03.

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Mohamed Zaid, Hagar, Khaled Abdel-ghaffar, Tarek Hessin El-bialy, Mamdouh Farid e Fatma Hamed M. El-demerdash. "Evaluation Of Regenerative Potential Of Pulp -derived Stem Cells And Gingival-derived Stem Cells In The Regeneration Of Periodontal Defects (experimental Study)". In Qatar Foundation Annual Research Conference Proceedings. Hamad bin Khalifa University Press (HBKU Press), 2014. http://dx.doi.org/10.5339/qfarc.2014.hbpp0741.

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Geti, Imbisaat. "Derivation of Hepatocyte Like Cells from Human Pluripotent Stem Cells in GMP Compliant Conditions". In Qatar Foundation Annual Research Conference Proceedings. Hamad bin Khalifa University Press (HBKU Press), 2016. http://dx.doi.org/10.5339/qfarc.2016.hbop2136.

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Abdelalim, Essam, Bushra Memon, Manale Karam e Sara Al-Khawaga. "Enhancement of differentiation of multipotent pancreatic β cell precursors from human embryonic stem cells". In Qatar Foundation Annual Research Conference Proceedings. Hamad bin Khalifa University Press (HBKU Press), 2018. http://dx.doi.org/10.5339/qfarc.2018.hbpp297.

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Parada, Luis F. "Abstract IA13: Glioma stem cells and cancer". In Abstracts: Third AACR International Conference on Frontiers in Basic Cancer Research - September 18-22, 2013; National Harbor, MD. American Association for Cancer Research, 2013. http://dx.doi.org/10.1158/1538-7445.fbcr13-ia13.

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Fouse, Shaun D., Raman P. Nagarajan, Jean Nakamura, C. David James, Susan Chang e Joseph F. Costello. "Abstract B44: Therapeutic response of primary glioblastoma cancer stem cells relative to patient-matched non-stem tumor cells". In Abstracts: Second AACR International Conference on Frontiers in Basic Cancer Research--Sep 14-18, 2011; San Francisco, CA. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.fbcr11-b44.

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Abdelalim, Essam, Ahmed Abdelaal, Bushra Memon, Yasmin Abu Aqel, Mohamed Bashir, Manjula Nandakumar e Shahrad Taheri. "Generation of induced pluripotent stem cells from insulin resistant Qatari patients". In Qatar Foundation Annual Research Conference Proceedings. Hamad bin Khalifa University Press (HBKU Press), 2018. http://dx.doi.org/10.5339/qfarc.2018.hbpd138.

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Rapporti di organizzazioni sul tema "Stem cells – Research"

Peterson, Daniel. Research Recruiting Endogenous Tissue Stem Cells to Repair Injury and Degeneration. Office of Scientific and Technical Information (OSTI), ottobre 2014. http://dx.doi.org/10.2172/1160139.

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Gupta, Shweta. The Revolution of Human Organoids in Cell Biology. Natur Library, ottobre 2020. http://dx.doi.org/10.47496/nl.blog.12.

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Organoids are a new research tool derived from human pluripotent or adult stem cells or somatic cells in vitro to form small, self-organizing 3-dimensional structures that simulate many of the functions of native organs

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Cao, Siyang, Yihao Wei, Tiantian Qi, Peng Liu, Yingqi Chen, Fei Yu, Hui Zeng e Jian Weng. Stem cell therapy for peripheral nerve injury: An up-to-date meta-analysis of 55 preclinical researches. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, ottobre 2022. http://dx.doi.org/10.37766/inplasy2022.10.0083.

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Review question / Objective: It has been the gold standard for decades to reconstruct a large peripheral nerve injury with a nerve autograft, and this remains true today as well. In addition to nerve autografts, biological conduits and vessels can also be applied. A fair amount of studies have examined the benefits of adding stem cells to the lumen of a nerve conduit. The aim of this meta-analysis was to summarize animal experiments related to the utilization of stem cells as a luminal additive when rebuilding a peripheral nerve injury using nerve grafts. Eligibility criteria: The inclusion criteria were as following: 1.Reconstruction of peripheral nerve injury; 2.Complete nerve transection with gap defect created; 3.Animal in-vivo models; 4.Experimental comparisons between nerve conduits containing and not containing one type of stem cell; 5.Functional testing and electrophysiology evaluations are performed. The exclusion criteria were as following: 1.Repair of central nervous system; 2.Nerve repair is accomplished by end-to-end anastomosis; 3.Animal models of entrapment injuries, frostbite, traction injuries and electric injuries; 4.Nerve conduits made from autologous epineurium; 5.Clinical trials, reviews, letters, conference papers, meta-analyses or commentaries; 6.Same studies have been published in different journals under the same or a different title.

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Halevy, Orna, Sandra Velleman e Shlomo Yahav. Early post-hatch thermal stress effects on broiler muscle development and performance. United States Department of Agriculture, gennaio 2013. http://dx.doi.org/10.32747/2013.7597933.bard.

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In broilers, the immediate post-hatch handling period exposes chicks to cold or hot thermal stress, with potentially harmful consequences to product quantity and quality that could threaten poultry meat marketability as a healthy, low-fat food. This lower performance includes adverse effects on muscle growth and damage to muscle structure (e.g., less protein and more fat deposition). A leading candidate for mediating the effects of thermal stress on muscle growth and development is a unique group of skeletal muscle cells known as adult myoblasts (satellite cells). Satellite cells are multipotential stem cells that can be stimulated to follow other developmental pathways, especially adipogenesis in lieu of muscle formation. They are most active during the first week of age in broilers and have been shown to be sensitive to environmental conditions and nutritional status. The hypothesis of the present study was that immediate post-hatch thermal stress would harm broiler growth and performance. In particular, growth characteristics and gene expression of muscle progenitor cells (i.e., satellite cells) will be affected, leading to increased fat deposition, resulting in long-term changes in muscle structure and a reduction in meat yield. The in vitro studies on cultured satellite cells derived from different muscle, have demonstrated that, anaerobic pectoralis major satellite cells are more predisposed to adipogenic conversion and more sensitive during myogenic proliferation and differentiation than aerobic biceps femoris cells when challenged to both hot and cold thermal stress. These results corroborated the in vivo studies, establishing that chronic heat exposure of broiler chicks at their first two week of life leads to impaired myogenicity of the satellite cells, and increased fat deposition in the muscle. Moreover, chronic exposure of chicks to inaccurate temperature, in particular to heat vs. cold, during their early posthatch periods has long-term effects of BW, absolute muscle growth and muscle morphology and meat quality. The latter is manifested by higher lipid and collagen deposition and may lead to the white striping occurrence. The results of this study emphasize the high sensitivity of muscle progenitor cells in the early posthatch period at a time when they are highly active and therefore the importance of rearing broiler chicks under accurate ambient temperatures. From an agricultural point of view, this research clearly demonstrates the immediate and long-term adverse effects on broiler muscling and fat formation due to chronic exposure to hot stress vs. cold temperatures at early age posthatch. These findings will aid in developing management strategies to improve broiler performance in Israel and the USA. BARD Report - Project4592 Page 2 of 29

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Coplin, David L., Shulamit Manulis e Isaac Barash. roles Hrp-dependent effector proteins and hrp gene regulation as determinants of virulence and host-specificity in Erwinia stewartii and E. herbicola pvs. gypsophilae and betae. United States Department of Agriculture, giugno 2005. http://dx.doi.org/10.32747/2005.7587216.bard.

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Gram-negative plant pathogenic bacteria employ specialized type-III secretion systems (TTSS) to deliver an arsenal of pathogenicity proteins directly into host cells. These secretion systems are encoded by hrp genes (for hypersensitive response and pathogenicity) and the effector proteins by so-called dsp or avr genes. The functions of effectors are to enable bacterial multiplication by damaging host cells and/or by blocking host defenses. We characterized essential hrp gene clusters in the Stewart's Wilt of maize pathogen, Pantoea stewartii subsp. stewartii (Pnss; formerly Erwinia stewartii) and the gall-forming bacterium, Pantoea agglomerans (formerly Erwinia herbicola) pvs. gypsophilae (Pag) and betae (Pab). We proposed that the virulence and host specificity of these pathogens is a function of a) the perception of specific host signals resulting in bacterial hrp gene expression and b) the action of specialized signal proteins (i.e. Hrp effectors) delivered into the plant cell. The specific objectives of the proposal were: 1) How is the expression of the hrp and effector genes regulated in response to host cell contact and the apoplastic environment? 2) What additional effector proteins are involved in pathogenicity? 3) Do the presently known Pantoea effector proteins enter host cells? 4) What host proteins interact with these effectors? We characterized the components of the hrp regulatory cascade (HrpXY ->7 HrpS ->7 HrpL ->7 hrp promoters), showed that they are conserved in both Pnss and Fag, and discovered that the regulation of the hrpS promoter (hrpSp) may be a key point in integrating apoplastic signals. We also analyzed the promoters recognized by HrpL and demonstrated the relationship between their composition and efficiency. Moreover, we showed that promoter strength can influence disease expression. In Pnss, we found that the HrpXY two-component signal system may sense the metabolic status of the bacterium and is required for full hrp gene expression in planta. In both species, acyl-homoserine lactone-mediated quorum sensing may also regulate epiphytic fitness and/or pathogenicity. A common Hrp effector protein, DspE/WtsE, is conserved and required for virulence of both species. When introduced into corn cells, Pnss WtsE protein caused water-soaked lesions. In other plants, it either caused cell death or acted as an Avr determinant. Using a yeast- two-hybrid system, WtsE was shown to interact with a number of maize signal transduction proteins that are likely to have roles in either programmed cell death or disease resistance. In Pag and Pab, we have characterized the effector proteins HsvG, HsvB and PthG. HsvG and HsvB are homologous proteins that determine host specificity of Pag and Pab on gypsophila and beet, respectively. Both possess a transcriptional activation domain that functions in yeast. PthG was found to act as an Avr determinant on multiple beet species, but was required for virulence on gypsophila. In addition, we demonstrated that PthG acts within the host cell. Additional effector genes have been characterized on the pathogenicity plasmid, pPATHₚₐg, in Pag. A screen for HrpL- regulated genes in Pnsspointed up 18 candidate effector proteins and four of these were required for full virulence. It is now well established that the virulence of Gram-negative plant pathogenic bacteria is governed by Hrp-dependent effector proteins. However; the mode of action of many effectors is still unresolved. This BARD supported research will significantly contribute to the understanding of how Hrp effectors operate in Pantoea spp. and how they control host specificity and affect symptom production. This may lead to novel approaches for genetically engineering plants resistant to a wide range of bacterial pathogens by inactivating the Hrp effectors with "plantabodies" or modifying their receptors, thereby blocking the induction of the susceptible response. Alternatively, innovative technologies could be used to interfere with the Hrp regulatory cascade by blocking a critical step or mimicking plant or quorum sensing signals.

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Granot, David, e Richard Amasino. Regulation of Senescence by Sugar Metabolism. United States Department of Agriculture, gennaio 2003. http://dx.doi.org/10.32747/2003.7585189.bard.

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Research objectives a. Analyze transgenic plants that undergo rapid senescence due to increased expression of hexokinase. b. Determine if hexokinase-induced senescence accelerates natural senescence using senescence specific promoters that drive expression of a reporter gene (GUS) and a cytokinin producing gene (IPT - isopentyl transferase). c. Isolate and analyze plant genes that suppress sugar-induced cell death (SICD) in yeast, genes that potentially are involved in programmed cell death and senescence in plants. Background to the topic Leaf senescence is a regulated process of programmed cell death (PCD) in which metabolites are recycled to other active parts of the plant. Senescence associated genes (SAGs) are expressed throughout leaf senescence. Sugar flux and metabolism is thought to playa fundamental regulatory role in senescence. We found that transgenic tomato plants with high hexokinase activity, the initial enzymatic step of sugar (hexose) metabolism, undergo rapid leaf senescence, directly correlated with hexokinase activity. These plants provide a unique opportunity to analyze the regulatory role of sugar metabolism in senescence, and its relation to cytokinin, a senescence-inhibiting hormone. In addition, we found that sugar induces programmed cells death of yeast cells in direct correlation to hexokinase activity. We proposed to use the sugar induced cell death (SICD) to isolate Arabidopsis genes that suppress SICD. Such genes could potentially be involved in senescence induced PCD in plants. Major conclusions The promoters of Arabidopsis senescence-associated genes, SAG12 and SAGI3, are expressed in senescing tomato leaves similar to their expression in Arabidopsis leaves, indicating that these promoters are good senescence markers for tomato plants. Increased hexokinase activity accelerated senescence and induced expression of pSAG12 and pSAG13 promoters in tomato plants, suggesting that sugar regulate natural senescence via hexokinase. Expression of IPT, a cytokinin producing gene, under pSAG12 and pSAG13 promoters, delayed senescence of tomato leaves. Yet, senescence accelerated by hexokinase was epistatic over cytokinin, indicating that sugar regulation of senescence is dominant over the senescence-inhibiting hormone. A gene designated SFP1, which is similar to the major super family monosaccharide transporters, is induced during leaf senescence in Arabidopsis and may be involved in sugar transport during senescence. Accordingly, adult leaves accumulate sugars that may accelerate hexokinase activity. Light status of the entire plant affects the senescence of individual leaves. When individual leaves are darkened, senescence is induced in the covered leaves. However, whole adult plant placed in darkness show delayed senescence. In a search for Arabidopsis genes that suppress SICD we isolated 8 cDNA clones which confer partial resistance to SICD. One of the clones encodes a vesicle associated membrane protein - VAMP. This is the first evidence that vesicle trafficking might be involved in cell death. Implications Increased hexokinase activity accelerates senescence. We hypothesized that, reduced hexokinase activity may delay senescence. Preliminary experiments using a hexokinase inhibitor support this possible implication. Currently we are analyzing various practical approaches to delay leaf senescence via hexokinase inhibition. .

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Blumwald, Eduardo, e Avi Sadka. Sugar and Acid Homeostasis in Citrus Fruit. United States Department of Agriculture, gennaio 2012. http://dx.doi.org/10.32747/2012.7697109.bard.

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Citrus fruit quality standards have been determined empirically, depending on species and on the particular growing regions. In general, the TSS (total soluble solids) to total acidity (TA) ratio determines whether citrus fruit can be marketed. Soluble sugars account for most of the TSS during harvest while TA is determined almost solely by the citric acid content, which reaches levels of 1-5% by weight in many cultivated varieties. Acid and sugar homeostasis in the fruit is critical for the management of existing cultivars, the development of new cultivars, the improvement of pre- and post-harvest strategies and the control of fruit quality and disorders. The current proposal (a continuation of a previous proposal) aimed at: (1) completing the citrus fruit proteome and metabolome, and establish a citrus fruit functional database, (2) further characterization of the control of fruit acidity by studying the regulation of key steps affecting citrate metabolism, and determine the fate of citrate during acid decline stage, and (3) Studying acid and sugar homeostasis in citrus fruits by characterizing transport mechanisms across membranes. These aims were completed as the following: (1) Our initial efforts were aimed at the characterization and identification of citric acid transporters in citrus juice cells. The identification of citrate transporters at the vacuole of the citrus juice cell indicated that the steady-state citrate cytosolic concentration and the action of the cytosolic aconitase were key elements in establishing the pH homeostat in the cell that regulates the metabolic shift towards carbon usage in the fruit during the later stages of fruit development. We focused on the action of aconitase, the enzyme mediating the metabolic use of citric acid in the cells, and identified processes that control carbon fluxes in developing citrus fruits that control the fruit acid load; (2) The regulation of aconitase, catalyzing a key step in citrate metabolism, was further characterized by using two inhibitors, citramalte and oxalomalte. These compounds significantly increased citrate content and reduced the enzyme’s activity. Metabolite profiling and changes of amino-acid metabolizing enzymes in oxalomalate- treated cells suggested that the increase in citrate, caused by aconitase inhibition, induces amino acid synthesis and the GABA shunt, in accordance with the suggested fate of citrate during the acid decline stage in citrus fruit. (3) We have placed a considerable amount of time on the development of a citrus fruit proteome that will serve to identify all of the proteins in the juice cells and will also serve as an aid to the genomics efforts of the citrus research community (validating the annotation of the fruit genes and the different ESTs). Initially, we identified more than 2,500 specific fruit proteins and were able to assign a function to more than 2,100 proteins (Katz et al., 2007). We have now developed a novel Differential Quantitative LC-MS/MS Proteomics Methodology for the identification and quantitation of key biochemical pathways in fruits (Katz et al., 2010) and applied this methodology to identify determinants of key traits for fruit quality (Katz et al., 2011). We built “biosynthesis maps” that will aid in defining key pathways associated with the development of key fruit quality traits. In addition, we constructed iCitrus (http://wiki.bioinformatics.ucdavis.edu/index.php/ICitrus), a “functional database” that is essentially a web interface to a look-up table that allows users to use functional annotations in the web to identify poorly annotated citrus proteins. This resource will serve as a tool for growers and field extension specialists.

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Philosoph-Hadas, Sonia, Peter B. Kaufman, Shimon Meir e Abraham H. Halevy. Inhibition of the Gravitropic Shoot Bending in Stored Cut Flowers Through Control of Their Graviperception: Involvement of the Cytoskeleton and Cytosolic Calcium. United States Department of Agriculture, dicembre 2005. http://dx.doi.org/10.32747/2005.7586533.bard.

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Original objectives: The basic goal of the present project was to study the mechanism involved in shoot graviperception and early transduction, in order to determine the sequence of events operating in this process. This will enable to control the entire process of gravity-induced differential growth without affecting vertical growth processes essential for development. Thus, several new postulated interactions, operating at the perception and early transduction stages of the signaling cascade leading to auxin-mediated bending, were proposed to be examined in snapdragon spikes and oat shoot pulvini, according to the following research goals: 1) Establish the role of amyloplasts as gravireceptors in shoots; 2) Investigate gravity-induced changes in the integrity of shoot actin cytoskeleton (CK); 3) Study the cellular interactions among actin CK, statoliths and cell membranes (endoplasmic reticulum - ER, plasma membrane - PM) during shoot graviperception; 4) Examine mediation of graviperception by modulations of cytosolic calcium - [Ca2+]cyt, and other second messengers (protein phosphorylation, inositol 1,4,5-trisphosphate - IP3). Revisions: 1) Model system: in addition to snapdragon (Antirrhinum majus L.) spikes and oat (Avena sativa) shoot pulvini, the model system of maize (Zea mays) primary roots was targeted to confirm a more general mechanism for graviperception. 2) Research topic: brassinolide, which were not included in the original plan, were examined for their regulatory role in gravity perception and signal transduction in roots, in relation to auxin and ethylene. Background to the topic: The negative gravitropic response of shoots is a complex multi-step process that requires the participation of various cellular components acting in succession or in parallel. Most of the long-lasting studies regarding the link between graviperception and cellular components were focused mainly on roots, and there are relatively few reports on shoot graviperception. Our previous project has successfully characterized several key events occurring during shoot bending of cut flowers and oat pulvini, including amyloplast displacement, hormonal interactions and differential growth analysis. Based on this evidence, the present project has focused on studying the initial graviperception process in flowering stems and cereal shoots. Major conclusions and achievements: 1) The actin and not the microtubule (MT) CK is involved in the graviperception of snapdragon shoots. 2) Gravisensing, exhibited by amyloplast displacement, and early transduction events (auxin redistribution) in the gravitropic response of snapdragon spikes are mediated by the acto-myosin complex. 3) MTs are involved in stem directional growth, which occurs during gravitropism of cut snapdragon spikes, but they are not necessary for the gravity-induced differential growth. 4) The role of amyloplasts as gravisensors in the shoot endodermis was demonstrated for both plant systems. 5) A gravity-induced increase in IP.

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Granot, David, e Noel Michelle Holbrook. Role of Fructokinases in the Development and Function of the Vascular System. United States Department of Agriculture, gennaio 2011. http://dx.doi.org/10.32747/2011.7592125.bard.

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Plant vascular tissues are superhighways whose development and function have profound implications for productivity, yield and stress response. Preliminary studies by the PI indicated that sugar metabolism mediated by fructokinases (FRKs) has a pronounced effect on the transport properties of the xylem. The goal of this research was to determine how the main fructokinase gene, FRK2, and the only plastidic fructokinase, FRK3, influence vascular development and physiology, emphasizing processes that occur at both the cellular and organismic level. We found that both genes are expressed in vascular tissues, but FRK3 is expressed primarily in vascular tissues of mature petioles. Vascular anatomy of plants with antisense suppression of FRK2 uncovered that FRK2 is necessary for xylem and phloem development, most likely due to its role in vascular cell-wall synthesis, and affects vascular development all over the plant. As a result, suppression of FRK2 reduced hydraulic conductivity of roots, stem and leaves and restricted sugar phloem transport. Vascular anatomy of plants with RNAi suppression of FRK3 uncovered that FRK3 is required for vascular development in mature petiole but its role is partially complemented by FRK2. Suppression of FRK3 combined with partial suppression of FRK2 had effects completely different from that of FRK2 suppression, resulting in wilting of mature leaves rather than young leaves of FRK2 suppressed plants, and decreased export of photoassimilates. This primary effect of FRK2 suppression on mature petioles had a secondary effect, reducing the hydraulic conductivity in roots and stem. The very fact that a plastidic fructokinase plays a role in vascular development is quite surprising and we are still seeking to uncover its metabolic mode-of-action. Yet, it is clear that these two fructokinases have different roles in the coordination between photosynthetic capacity and vascular development. We have started analyzing the role of the last third FRK, FRK1, and discovered that it is also expressed exclusively in vascular tissues. It appears therefore, that all FRKs studied here are involved in vascular development. An interesting unexpected outcome of this study was the connection of FRK2 with hormonal regulation of vascular development, most likely auxin. This observation together with the yet to be solved questions on the exact roles of FRK3 are the subjects of our current efforts.

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Gafny, Ron, A. L. N. Rao e Edna Tanne. Etiology of the Rugose Wood Disease of Grapevine and Molecular Study of the Associated Trichoviruses. United States Department of Agriculture, settembre 2000. http://dx.doi.org/10.32747/2000.7575269.bard.

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Rugose wood is a complex disease of grapevines, characterized by modification of the woody cylinder of affected vines. The control of rugose wood is based on the production of healthy propagation material. Detection of rugose wood in grapevines is difficult and expensive: budwood from tested plants is grafted onto sensitive Vitis indicators and the appearance of symptoms is monitored for 3 years. The etiology of rugose wood is complex and has not yet been elucidated. Several elongated clostero-like viruses are consistently found in affected vines; one of them, grapevine virus A (GVA), is closely associated with Kober stem grooving, a component of the rugose wood complex. GVA has a single-stranded RNA genome of 7349 nucleotides, excluding a polyA tail at the 3' terminus. The GVA genome includes five open reading frames (ORFs 1-5). ORF 4, which encodes for the coat protein of GVA, is the only ORF for which the function was determined experimentally. The original objectives of this research were: 1- To produce antisera to the structural and non-structural proteins of GVA and GVB and to use these antibodies to establish an effective detection method. 2- Develop full length infectious cDNA clones of GVA and GVB. 3- Study the roll of GVA and GVB in the etiology of the grapevine rugose wood disease. 4- Determine the function of Trichovirus (now called Vitivirus) encoded genes in the virus life cycle. Each of the ORFs 2, 3, 4 and 5 genes of GVA were cloned and expressed in E. coli and used to produce antisera. Both the CP (ORF 4) and the putative MP (ORF 3) were detected with their corresponding antisera in-GVA infected N. benthamiana and grapevine. The MP was first detected at an early stage of the infection, 6-12 h after inoculation, and the CP 2-3 days after inoculation. The MP could be detected in GVA-infected grapevines that tested negative for CP, both with CP antiserum and with a commercially available ELISA kit. Antisera to ORF 2 and 5 encoded proteins could react with the recombinant proteins but failed to detect both proteins in GVA infected plants. A full-length cDNA clone of grapevine virus A (GVA) was constructed downstream from the bacteriophage T7 RNA polymerase promoter. Capped in vitro transcribed RNA was infectious in N. benthamiana and N. clevelandii plants. Symptoms induced by the RNA transcripts or by the parental virus were indistinguishable. The infectivity of the in vitro-transcribed RNA was confirmed by serological detection of the virus coat and movement proteins and by observation of virions by electron microscopy. The full-length clone was modified to include a gus reporter gene and gus activity was detected in inoculated and systemic leaves of infected plants. Studies of GVA mutants suggests that the coat protein (ORF 4) is essential for cell to cell movement, the putative movement protein (ORF 3) indeed functions as a movement protein and that ORF 2 is not required for virus replication, cell to cell or systemic movement. Attempts to infect grapevines by in-vitro transcripts, by inoculation of cDNA construct in which the virus is derived by the CaMV 35S promoter or by approach grafting with infected N. benthamiana, have so far failed. Studies of the subcellular distribution of GFP fusion with each of ORF 2, 3 and 4 encoded protein showed that the CP fusion protein accumulated as a soluble cytoplasmatic protein. The ORF 2 fusion protein accumulated in cytoplasmatic aggregates. The MP-GFP fusion protein accumulated in a large number of small aggregates in the cytoplasm and could not move from cell to cell. However, in conditions that allowed movement of the fusion protein from cell to cell (expression by a PVX vector or in young immature leaves) the protein did not form cytoplasmatic aggregates but accumulated in the plasmodesmata.

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