Relevant bibliographies by topics / Cell biology research

Academic literature on the topic 'Cell biology research'

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

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Journal articles on the topic "Cell biology research"

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Fields, S. "CELL BIOLOGY: Whither Model Organism Research?" Science 307, no. 5717 (March 25, 2005): 1885–86. http://dx.doi.org/10.1126/science.1108872.

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Choi, Yongmun, and Tae-gyu Nam. "Chemical biology in stem cell research." Archives of Pharmacal Research 35, no. 2 (February 2012): 281–97. http://dx.doi.org/10.1007/s12272-012-0208-6.

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Gordon, MY, F. Dazzi, SB Marley, JL Lewis, D. Nguyen, FH Grand, RJ Davidson, and JM Goldman. "Cell biology of CML cells." Leukemia 13, S1 (April 1999): S65—S71. http://dx.doi.org/10.1038/sj.leu.2401281.

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Gazdar, Adi F. "Cell biology and molecular biology of small cell and non-small cell lung cancer." Current Opinion in Oncology 2, no. 2 (April 1990): 321–27. http://dx.doi.org/10.1097/00001622-199004000-00013.

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Chicurel, M. "CELL BIOLOGY: Cell Migration Research Is on the Move." Science 295, no. 5555 (January 25, 2002): 606–9. http://dx.doi.org/10.1126/science.295.5555.606.

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Pontén, J. "Cell biology of precancer." European Journal of Cancer 37 (September 2001): 97–113. http://dx.doi.org/10.1016/s0959-8049(01)00277-5.

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Long, P. M., U. V. Wesley, D. M. Jaworski, M. Rana, T. R. Kiehl, K. So, P. Gould, et al. "Cell Biology and Signaling." Neuro-Oncology 12, Supplement 4 (October 21, 2010): iv7—iv25. http://dx.doi.org/10.1093/neuonc/noq116.s2.

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Agarwal, M., R. Nitta, S. Dovat, G. Li, H. Arita, Y. Narita, S. Fukushima, et al. "CELL BIOLOGY AND SIGNALING." Neuro-Oncology 15, suppl 3 (November 1, 2013): iii12—iii31. http://dx.doi.org/10.1093/neuonc/not174.

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Ku, Ja-Lok, and Jae-Gahb Park. "Biology of SNU Cell Lines." Cancer Research and Treatment 37, no. 1 (2005): 1. http://dx.doi.org/10.4143/crt.2005.37.1.1.

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Bebber, Christina M., Fabienne Müller, Laura Prieto Clemente, Josephine Weber, and Silvia von Karstedt. "Ferroptosis in Cancer Cell Biology." Cancers 12, no. 1 (January 9, 2020): 164. http://dx.doi.org/10.3390/cancers12010164.

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A major hallmark of cancer is successful evasion of regulated forms of cell death. Ferroptosis is a recently discovered type of regulated necrosis which, unlike apoptosis or necroptosis, is independent of caspase activity and receptor-interacting protein 1 (RIPK1) kinase activity. Instead, ferroptotic cells die following iron-dependent lipid peroxidation, a process which is antagonised by glutathione peroxidase 4 (GPX4) and ferroptosis suppressor protein 1 (FSP1). Importantly, tumour cells escaping other forms of cell death have been suggested to maintain or acquire sensitivity to ferroptosis. Therefore, therapeutic exploitation of ferroptosis in cancer has received increasing attention. Here, we systematically review current literature on ferroptosis signalling, cross-signalling to cellular metabolism in cancer and a potential role for ferroptosis in tumour suppression and tumour immunology. By summarising current findings on cell biology relevant to ferroptosis in cancer, we aim to point out new conceptual avenues for utilising ferroptosis in systemic treatment approaches for cancer.
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Dissertations / Theses on the topic "Cell biology research"

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Chen, Luxi. "Human Innate Lymphoid Cell Biology and Development." The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1551901769401192.

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Li, Qiming. "IL-1 Receptor Biology: Promoter Complex and Cell Type Specific Function." The Ohio State University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=osu1280861203.

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Calderone, Carli E. "Stem Cell Research: Science Education and Outreach." Miami University Honors Theses / OhioLINK, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=muhonors1268751337.

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Alvarado, Cabellos Alvaro Gonzalo. "Targeting Stem Cell Pathways in Glioblastoma." Case Western Reserve University School of Graduate Studies / OhioLINK, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=case1459189591.

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Jaime-Ramirez, Alena Cristina. "HER2 and Folate Receptor Targeted Therapy is Enhanced by NK Cell-Activating Cytokines." The Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1364465780.

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Wray, Jason Patrick. "Characterisation of a novel culture condition for the establishment and maintenance of mouse embryonic stem cells and implications for the mechanisms of self-renewal." Thesis, University of Edinburgh, 2009. http://hdl.handle.net/1842/3215.

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Pluripotency is defined as the ability of a cell to give rise to all the cell types of the adult organism. In vivo this property is possessed transiently by the cells of the epiblast in the developing embryo but it can be maintained indefinitely by deriving embryonic stem (ES) cells. How the pluripotent state is established in the cells of the early embryo and how it is ‘captured’ and maintained in the form of ES cells is a fascinating question for biology with practical implications. It is hoped that ES cells will be of use in biomedical research and cell replacement therapy. Our understanding of their biology and our ability to manipulate the cells in vitro will be of great importance if these hopes are to be realised. The starting point for the work presented in this thesis was the development of a novel culture condition for the derivation and maintenance of mouse ES cells (Q-L. Ying and J. Nichols). The media is formed by the addition of three small molecule inhibitors to a previously described serum-free media, N2B27, and is termed 3i (three inhibitors).The inhibitors are SU5402, PD184352 and CHIRON99021, and they inhibit the FGF receptor, mitogen activated protein/extracellular signalregulated kinase (ERK) kinase (MEK), and glycogen synthase kinase 3 (GSK3) respectively. I attempt to further our understanding of pluripotency and self-renewal in ES cells by genetic and biochemical examination of ES cells cultured in 3i. Analysis of intracellular signalling pathways together with descriptions of genetic mutants for the targets of the inhibitors validates the mode of action and the specificity of the three inhibitors. Self-renewal of mouse ES cells is considered dependent on activation of STAT3 through provision of the cytokine leukaemia inhibitory factor (LIF). I demonstrate unequivocally that this pathway is not required for self-renewal in 3i by characterising Stat3-null ES cells. Further experiments reveal that preventing activation of ERK downstream of the growth factor FGF4, produced by the ES cells themselves, is key to preventing differentiation. Pleiotropic effects of GSK3 inhibition are observed and candidate GSK3 targets with known or predicted effects on self-renewal are investigated as potential downstream effectors. I propose that activation of canonical Wnt signalling, together with a global derepression of biosynthetic capacity, mediate the pro-self-renewal effects of GSK3 inhibition. The description of culture conditions that function independently of signalling pathways previously thought essential for self-renewal provides fresh insight into the nature of ES cell self-renewal and the relationship of ES cells to the pluripotent cells of the developing embryo. There are practical implications for ES cell biology as there is reason to hope that the new conditions will translate more readily to other mammalian species to facilitate the derivation of ES cells and will provide an optimal platform for differentiation of ES cells into somatic cell types of interest.
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Chakraborty, Rikhia. "Homeostatic Regulation of Interleukin-4-Mediated Cell Signaling." Cleveland State University / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=csu1258663290.

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Sprague, Leslee W. "Dendritic Cell Culture With 2D and 3D Collagen Substrates." Ohio University / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1311616312.

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Hsu, En-Chi. "Targeting ILK to eliminate IL-6 induced cancer stem cell phenotype." The Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1397560686.

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Nordén, Johan. "Assessment of methods for microRNA isolation, microRNA amplification, and development of a normalization strategy for sepsis biomarker research." Thesis, Högskolan i Skövde, Institutionen för biovetenskap, 2020. http://urn.kb.se/resolve?urn=urn:nbn:se:his:diva-18442.

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Sepsis, defined by organ dysfunction caused by an adverse immune response of the host to an infection, comes with considerable cost in human lives and as a substantial burden financially. Significant upgrades have been made over the past two decades when diagnosing and treating sepsis but still with room for improvements. Early detection is a cornerstone in the fight against sepsis, and the focus on strengthening diagnostics is in the forefront of modern research. The implementation of biomarkers may be the path of progression in this objective. This study aimed at establishing procedural foundations when using microRNAs as potential biomarkers. The study conducted looked at: (1) Isolation procedure, of microRNA from human plasma, of three kits: Total RNA Purification Kit (Norgen Biotech), miRNAeasy Serum/Plasma Kit (Qiagen), and miRNeasy Serum/Plasma Advanced Kit (Qiagen). (2) Amplification of miRNA through two Reverse Transcription Quantitative PCR methods: Two-tailed RT-qPCR (TATAA Biocenter), and miRCURY LNA miRNA PCR (Qiagen). (3) Developing a normalization strategy by identifying miRNA reference targets in a geNorm pilot experiment. Qubit analysis revealed that the two isolation kits from Qiagen performed similar, and better that the Norgen kit. The Two-tailed RT-qPCR failed to amplify miRNA samples, whereas the miRCURY LNA miRNA PCR showed consistent amplification across samples with a high call rate. The geNorm analysis concluded that hsa-miR-425-5p and hsa-miR-93-5p was the optimal reference target set. The study demonstrated that the isolation kits from Qiagen coupled with the miRCURY LNA miRNA PCR is a viable option for future miRNA biomarker studies.
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Books on the topic "Cell biology research"

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Foster, Anne H. Encyclopedia of cell biology research. Hauppauge, N.Y: Nova Science Publisher's, 2011.

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Shartava, Tsisana. DNA research, genetics, and cell biology. Hauppauge, N.Y: Nova Science Publishers, 2011.

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service), ScienceDirect (Online, ed. Essentials of stem cell biology. 2nd ed. Amsterdam: Elsevier, 2009.

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R, Balkwill Frances, ed. Cytokine cell biology: A practical approach. 3rd ed. New York: Oxford University Press, 2000.

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Lin, Yunfeng, and Ronghui Zhou, eds. Advances in Nanomaterials-based Cell Biology Research. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-16-2666-1.

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Stem cell biology in health and disease. Dordrecht: Springer, 2009.

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Gheorghe, Benga, and Tager J. M, eds. Biomembranes: Basic and medical research. Berlin: Springer-Verlag, 1988.

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Essential zebrafish methods: Cell and developmental biology. Amsterdam: Elsevier, 2009.

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Essential stem cell methods. London: Academic Press, 2009.

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Zhao, Robert Chunhua. Essentials of mesenchymal stem cell biology and its clinical translation. Dordrecht: Springer, 2013.

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Book chapters on the topic "Cell biology research"

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Fagan, Melinda Bonnie. "Stem cell biology." In The Matrix of Stem Cell Research, 111–30. Milton Park, Abingdon, Oxon ; New York, NY : Routledge, 2020.: Routledge, 2019. http://dx.doi.org/10.4324/9781315104386-8.

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Sun, Zhen, Jing Zhao, Hua Yu, Chenyang Zhang, Hu Li, Zhongda Zeng, and Jin Zhang. "Metabolomics in Stem Cell Biology Research." In Computational Stem Cell Biology, 321–30. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9224-9_15.

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Saigusa, Ryosuke, Christopher P. Durant, Vasantika Suryawanshi, and Klaus Ley. "Single-Cell in Research." In Methods in Molecular Biology, 765–78. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-1924-7_46.

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Seifertová, Daniela, Petr Klíma, Markéta Pařezová, Jan Petrášek, Eva Zažímalová, and Zdeněk Opatrný. "Plant Cell Lines in Cell Morphogenesis Research." In Methods in Molecular Biology, 215–29. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-643-6_18.

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Huang, Rui, and Shuyan Wu. "Cell Models in Autophagy Research." In Autophagy: Biology and Diseases, 311–32. Singapore: Springer Singapore, 2021. http://dx.doi.org/10.1007/978-981-16-2830-6_14.

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Seifinejad, Ali. "Advances in Induced Pluripotent Stem Cell Biology." In Advances in Stem Cell Research, 67–84. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-940-2_5.

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McCarthy, Peter, and Quenten Schwarz. "Proteomics in Neural Crest Cell Research." In Methods in Molecular Biology, 153–65. New York, NY: Springer New York, 2019. http://dx.doi.org/10.1007/978-1-4939-9412-0_12.

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Willats, William G. T., Lesley McCartney, and J. Paul Knox. "Pectin Cell Biology: Complexity in Context." In Advances in Pectin and Pectinase Research, 147–57. Dordrecht: Springer Netherlands, 2003. http://dx.doi.org/10.1007/978-94-017-0331-4_11.

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Kosaka, Nobuyoshi, Yusuke Yoshioka, Keitaro Hagiwara, Naoomi Tominaga, and Takahiro Ochiya. "Functional Analysis of Exosomal MicroRNA in Cell–Cell Communication Research." In Methods in Molecular Biology, 1–10. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-453-1_1.

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Totey, Swapnil, Rajarshi Pal, Murali Krishna Mamidi, Vijayendran Govindasamy, and Satish Totey. "Stem Cell Biology and Application." In Applications of Flow Cytometry in Stem Cell Research and Tissue Regeneration, 75–89. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2012. http://dx.doi.org/10.1002/9780470631119.ch6.

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Conference papers on the topic "Cell biology research"

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Gamba, Gerardo, Daniela Riccardi, James C. Williams, Andrew P. Evan, James E. Lingeman, and James A. McAteer. "Cell Biology of Thiazide Bone Effects." In RENAL STONE DISEASE 2: 2nd International Urolithiasis Research Symposium. AIP, 2008. http://dx.doi.org/10.1063/1.2998060.

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Safitri, Nur Lina, Siti Zubaidah, Fatchur Rohman, and Sulisetijono Sulisetijono. "Online learning of cell biology from biology students’ perspectives." In THE 5TH INTERNATIONAL CONFERENCE ON MATHEMATICS AND SCIENCE EDUCATION (ICoMSE) 2021: Science and Mathematics Education Research: Current Challenges and Opportunities. AIP Publishing, 2023. http://dx.doi.org/10.1063/5.0112426.

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Truong, Danh D., Emre Arslan, Margarita Divenko, Sandhya Krishnan, Alexander Lazar, Kunal Rai, and Joseph Ludwig. "Unraveling the Biology of Desmoplastic Small Round Cell Tumor." In Leading Edge of Cancer Research Symposium. The University of Texas at MD Anderson Cancer Center, 2022. http://dx.doi.org/10.52519/00091.

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Clark, Greg. "Freshman Research Initiative: Teaching plant cell signaling through undergraduate research." In ASPB PLANT BIOLOGY 2020. USA: ASPB, 2020. http://dx.doi.org/10.46678/pb.20.240869.

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Christ, Kevin V., and Kevin T. Turner. "Hydrodynamically-Confined Microflows for Cell Adhesion Strength Measurement." In ASME 2010 First Global Congress on NanoEngineering for Medicine and Biology. ASMEDC, 2010. http://dx.doi.org/10.1115/nemb2010-13007.

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Cell adhesion plays a fundamental role in numerous physiological and pathological processes, and measurements of the adhesion strength are important in fields ranging from basic cell biology research to the development of implantable biomaterials. Our group and others have recently demonstrated that microfluidic devices offer advantages for characterizing the adhesion of cells to protein-coated surfaces [1,2]. Microfluidic devices offer many advantages over conventional assays, including the ability to apply high shear stresses in the laminar regime and the opportunity to directly observe cell behavior during testing. However, a key disadvantage is that such assays require cells to be cultured inside closed microchannels. Assays based on closed channels restrict the types of surfaces that can be examined and are not compatible with many standard techniques in cell biology research. Furthermore, while techniques for cell culture in microchannels have become common, maintaining the viability of certain types of cells in channels remains a challenge.
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Tanabashi, Sayuri. "CELL BIOLOGY EDUCATION WITH ADVANCED 3D TECHNOLOGIES FOR K-12 STUDENTS." In 15th annual International Conference of Education, Research and Innovation. IATED, 2022. http://dx.doi.org/10.21125/iceri.2022.0087.

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Papkovsky, D. B. "BS1.2 - Phosphorescence based oxygen sensors - essential tools for cell biology and life science research." In 17th International Meeting on Chemical Sensors - IMCS 2018. AMA Service GmbH, Von-Münchhausen-Str. 49, 31515 Wunstorf, Germany, 2018. http://dx.doi.org/10.5162/imcs2018/bs1.2.

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Xiao, Hua, Jianwen Xiong, Jiming Wu, and Zhenxi Zhang. "Experimental research on ALA-PDT destruction of leukaemie cell HL60." In Third International Conference on Photonics and Imaging in Biology and Medicine, edited by Qingming Luo, Valery V. Tuchin, Min Gu, and Lihong V. Wang. SPIE, 2003. http://dx.doi.org/10.1117/12.546146.

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Tanabashi, Sayuri. "Work-in-Progress-Plant Cell Biology Education Using Advanced 3D Technologies for K-12 Students." In 2021 7th International Conference of the Immersive Learning Research Network (iLRN). IEEE, 2021. http://dx.doi.org/10.23919/ilrn52045.2021.9459333.

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Souchelnytskyi, Serhiy. "Systemic properties of Carcinogenesis: Lessons from studies on the Earth and in the Space." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0118.

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proteins and genes act in coordinated ways, and their relations are visualized as networks. Networks are more accurate descriptions of cancer regulatory mechanisms, in comparison to lists of oncogenes and tumor suppressors. To extract essential regulators (nodes) and connections (edges), interrogations of these networks are performed, e.g. cancer cells are subjected to different treatments. Interrogations force cancer cells to engage nodes and edges essential for maintaining cancer properties, i.e. drivers, and nonessential followers. The challenge is to discriminate which of the mechanisms drive tumorigenesis, and which are followers. Interrogation of cancer cells under variable g-forces is the treatment to which cancer cells are not normally exposed. Therefore, low (weightlessness) and high (acceleration) g-forces may trigger responses, which may differ in part of followers from responses on the Earth, but still engage carcinogenesis-essential drivers nodes and edges. Methodology: Experimental interrogation of human cancer cells to generate carcinogenesis-related regulatory networks was performed by using proteomics, cell biology, biochemistry, immunohistochemistry and bioinformatics tools. We used also reported datasets deposited in various databases. These networks were analyzed with algorithms to extract drivers of carcinogenesis. Results: Systemic analysis of human breast carcinogenesis has shown mechanisms of engagement of all known cancer hallmarks. Moreover, novel hallmarks have emerged, e.g. involvement of mechanisms of virus-cell interaction and RNA/miR processing. The breast cancer networks are rich, with >6,000 involved proteins and genes. The richness of the networks may explain many clinical observations, e.g. personalized response to treatments. Systemic analysis highlighted novel opportunities for treatment of cancer, by identifying key nodes of known and novel hallmark mechanisms. Systemic properties of the cancer network provides an opportunity to study compensatory mechanisms. These compensatory mechanisms frequently contribute to development of resistance to treatment. These mechanisms will be discussed. Cancer cells are not “wired” to function in weightlessness. The cells would have to adapt. This adaptation will include preserving mechanisms driving carcinogenesis, in addition to the space-only-related adaptation. Key carcinogenesis regulators in the space would be the same as on the Earth, while “passenger”-mechanisms would differ. Systems biology allows integration of a space- and the Earth-data, and would extract key regulators, and, subsequently lead to better diagnostic. Conclusion: Systemic analysis of carcinogenesis studies with different ways of interrogation delivered better diagnostic and novel modalities of treatment.
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Reports on the topic "Cell biology research"

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Zatz, Martin. Gordon Research Conference On Pineal Cell Biology. Fort Belvoir, VA: Defense Technical Information Center, July 1992. http://dx.doi.org/10.21236/ada264840.

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Gupta, Shweta. The Revolution of Human Organoids in Cell Biology. Natur Library, October 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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Jarron, Matthew, Amy R. Cameron, and James Gemmill. Dundee Discoveries Past and Present. University of Dundee, November 2020. http://dx.doi.org/10.20933/100001182.

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A series of self-guided walking tours through pioneering scientific research in medicine, biology, forensics, nursing and dentistry from the past to the present. Dundee is now celebrated internationally for its pioneering work in medical sciences, in particular the University of Dundee’s ground-breaking research into cancer, diabetes, drug development and surgical techniques. But the city has many more amazing stories of innovation and discovery in medicine and biology, past and present, and the three walking tours presented here will introduce you to some of the most extraordinary. Basic information about each topic is presented on this map, but you will ­find more in-depth information, images and videos on the accompanying website at uod.ac.uk/DundeeDiscoveriesMap For younger explorers, we have also included a Scavenger Hunt – look out for the cancer cell symbols on the map and see if you can ­find the various features listed along the way!
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Kang, Jing, Jun Zhang, Zongsheng Tian, Ye Xu, Jiangbi Li, and Mingxina Li. The efficacy and safety of immune-checkpoint inhibitor plus chemotherapy versus chemotherapy for non-small cell lung cancer: an updated systematic review and meta-analysis. INPLASY - International Platform of Registered Systematic Review and Meta-analysis Protocols, May 2022. http://dx.doi.org/10.37766/inplasy2022.5.0156.

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Review question / Objective: Population: histologically confirmed advanced NSCLC patients; Intervention: received immune-checkpoint inhibitor plus chemotherapy; Comparison:received chemotherapy; Outcome: reported OS, PFS, ORR and TRAEs; Study design: RCT. Condition being studied: Lung cancer is the primary cause of cancer-related deaths, with an estimated 2.20 million new cases and 1.79 million deaths every year, and 85% of all primary lung cancers are non-small cell lung cancer. Eligibility criteria: Studies were considered eligible if they met the following criteria: (1) being an randomized controlled trial published in English, (2) histologically confirmed advanced NSCLC patients, (3) reported OS, PFS, ORR and TRAEs, (4) the intervention group received immune-checkpoint inhibitor plus chemotherapy, while the control group received chemotherapy, (5) When numerous papers reporting the same trial were found, the most current or most complete publications were chosen. The following were the exclusion criteria: (1) duplicate articles, (2) reviews, meta-analyses, case reports, editorials and letters, (3) molecular biology or animal research, (4) retrospective or prospective observational cohort studies.
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Palmer, Guy, Varda Shkap, Wendy Brown, and Thea Molad. Control of bovine anaplasmosis: cytokine enhancement of vaccine efficacy. United States Department of Agriculture, March 2007. http://dx.doi.org/10.32747/2007.7695879.bard.

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Anaplasmosis an arthropod-born disease of cattle caused by the rickettsia Anaplasma marginale and is an impediment to efficient production of healthy livestock in both Israel and the United States. Currently the only effective vaccines are derived from the blood of infected cattle. The risk of widespread transmission of both known and newly emergent pathogens has prevented licensure of live blood-based vaccines in the U.S. and is a major concern for their continued use in Israel. Consequently development of a safe, effective vaccine is a high priority. In this collaborative project we focused on two approaches to vaccine development. The first focused o n improving antigen delivery to livestock and specifically examined how DNA vaccines could be improved to enhance priming and expansion of the immune response. This research resulted in development and testing of two novel vaccine delivery systems--one that targeted antigen spread among dendritic cells (the key cell in priming immune responses and a follow-on construct that also specifically targeted antigen to the endosomal-lysosomal compartment the processing organelle within the dendritic cell that directs vaccine antigen to the MHC class ll-CD4* T cell priming pathway). The optimized construct targeting vaccine antigen to the dendritic cell MHC class II pathway was tested for ability to prime A. marginale specific immune responses in outbred cattle. The results demonstrated both statistically significant effects of priming with a single immunization, continued expansion of the primary immune response including development of high affinity lgG antibodies and rapid recall of the memory response following antigen challenge. This portion of the study represented a significant advance in vaccine delivery for livestock. Importantly the impact of these studies is not limited to A. marginale a s the targeting motifs are optimized for cattle and can be adapted to other cattle vaccinations by inserting a relevant pathogen-specific antigen. The second approach (which represented an addition to the project for which approval was requested as part of the first annual report) was a comparative approach between A . marginale and the Israel A . centrale vaccines train. This addition was requested as studies on Major Surface Protein( MSP)- 2 have shown that this antigen is highly antigenically variable and presented solely as a "static vaccine" antigen does not give cross-strain immunity. In contrast A. . centrale is an effective vaccine which Kimron Veterinary institute has used in the field in Israel for over 50 years. Taking advantage of this expertise, a broad comparison of wild type A. marginale and vaccine strain was initiated. These studies revealed three primary findings: i) use of the vaccine is associated with superinfection, but absence of clinical disease upon superinfection with A. marginale; ii) the A. centrale vaccine strain is not only less virulent but transmission in competent in Dermacentor spp. ticks; and iii) some but not all MSPs are conserved in basic orthologous structure but there are significant polymorphisms among the strains. These studies clearly indicated that there are statistically significant differences in biology (virulence and transmission) and provide a clear path for mapping of biology with the genomes. Based on these findings, we initiated complete genome sequencing of the Israel vaccine strain (although not currently funded by BARD) and plant to proceed with a comparative genomics approach using already sequenced wild-type A. marginale. These findings and ongoing collaborative research tie together filed vaccine experience with new genomic data, providing a new approach to vaccine development against a complex pathogen.
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6

Sessa, Guido, and Gregory Martin. MAP kinase cascades activated by SlMAPKKKε and their involvement in tomato resistance to bacterial pathogens. United States Department of Agriculture, January 2012. http://dx.doi.org/10.32747/2012.7699834.bard.

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The research problem: Pseudomonas syringae pv. tomato (Pst) and Xanthomonas campestrispv. vesicatoria (Xcv) are the causal agents of tomato bacterial speck and spot diseases, respectively. These pathogens colonize the aerial parts of the plant and cause economically important losses to tomato yield worldwide. Control of speck and spot diseases by cultural practices or chemicals is not effective and genetic sources of resistance are very limited. In previous research supported by BARD, by gene expression profiling we identified signaling components involved in resistance to Xcvstrains. Follow up experiments revealed that a tomato gene encoding a MAP kinase kinase kinase (MAPKKKe) is required for resistance to Xcvand Pststrains. Goals: Central goal of this research was to investigate the molecular mechanisms by which MAPKKKεand associated MAP kinase cascades regulate host resistance. Specific objectives were to: 1. Determine whether MAPKKKεplays a broad role in defense signaling in plants; 2. Identify components of MAP kinase cascades acting downstream of MAPKKKε; 3. Determine the role of phosphorylation-related events in the function of MAPKKKε; 4. Isolate proteins directly activated by MAPKKKε-associatedMAPK modules. Our main achievements during this research program are in the following major areas: 1. Characterization of MAPKKKεas a positive regulator of cell death and dissection of downstream MAP kinase cascades (Melech-Bonfil et al., 2010; Melech-Bonfil and Sessa, 2011). The MAPKKKεgene was found to be required for tomato resistance to Xcvand Pstbacterial strains and for hypersensitive response cell death triggered by different R gene/effector gene pairs. In addition, overexpression analysis demonstrated that MAPKKKεis a positive regulator of cell death, whose activity depends on an intact kinase catalytic domain. Epistatic experiments delineated a signaling cascade downstream of MAPKKKεand identified SIPKK as a negative regulator of MAPKKKε-mediated cell death. Finally, genes encoding MAP kinase components downstream of MAPKKKεwere shown to contribute to tomato resistance to Xcv. 2. Identification of tomato proteins that interact with MAPKKKεand play a role in plant immunity (Oh et al., 2011). We identified proteins that interact with MAPKKKε. Among them, the 14-3-3 protein TFT7 was required for cell death mediated by several R proteins. In addition, TFT7 interacted with the MAPKK SlMKK2 and formed homodimersin vivo. Thus, TFT7 is proposed to recruit SlMKK2 and MAPKKK client proteins for efficient signal transfer. 3. Development of a chemical genetic approach to identify substrates of MAPKKKε-activated MAP kinase cascades (Salomon et al., 2009, 2011). This approach is based on engineering the kinase of interest to accept unnatural ATP analogs. For its implementation to identify substrates of MAPKKKε-activated MAP kinase modules, we sensitized the tomato MAP kinase SlMPK3 to ATP analogs and verified its ability to use them as phosphodonors. By using the sensitized SlMPK3 and radiolabeled N6(benzyl)ATP it should be possible to tag direct substrates of this kinase. 4. Development of methods to study immunity triggered by pathogen-associated molecular patterns (PAMPs) in tomato and N. benthamiana plants (Kim et al., 2009; Nguyen et al. 2010). We developed protocols for measuring various PTI-associatedphenotypes, including bacterial populations after pretreatment of leaves with PAMPs, induction of reporter genes, callose deposition at the cell wall, activation of MAP kinases, and a luciferase-based reporter system for use in protoplasts. Scientific and agricultural significance: Our research activities discovered and characterized a signal transduction pathway mediating plant immunity to bacterial pathogens. Increased understanding of molecular mechanisms of immunity will allow them to be manipulated by both molecular breeding and genetic engineering to produce plants with enhanced natural defense against disease. In addition, we successfully developed new biochemical and molecular methods that can be implemented in the study of plant immunity and other aspects of plant biology.
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7

Dickman, Martin B., and Oded Yarden. Genetic and chemical intervention in ROS signaling pathways affecting development and pathogenicity of Sclerotinia sclerotiorum. United States Department of Agriculture, July 2015. http://dx.doi.org/10.32747/2015.7699866.bard.

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Abstract: The long-term goals of our research are to understand the regulation of sclerotial development and pathogenicity in S. sclerotior11111. The focus in this project was on the elucidation of the signaling events and environmental cues involved in the regulation of these processes, utilizing and continuously developing tools our research groups have established and/or adapted for analysis of S. sclerotiorum, Our stated objectives: To take advantage of the recent conceptual (ROS/PPs signaling) and technical (amenability of S. sclerotiorumto manipulations coupled with chemical genomics and next generation sequencing) developments to address and extend our fundamental and potentially applicable knowledge of the following questions concerning the involvement of REDOX signaling and protein dephosphorylation in the regulation of hyphal/sclerotial development and pathogenicity of S. sclerotiorum: (i) How do defects in genes involved in ROS signaling affect S. sclerotiorumdevelopment and pathogenicity? (ii) In what manner do phosphotyrosinephosphatases affect S. sclerotiorumdevelopment and pathogenicity and how are they linked with ROS and other signaling pathways? And (iii) What is the nature of activity of newly identified compounds that affect S. sclerotiori,111 growth? What are the fungal targets and do they interfere with ROS signaling? We have met a significant portion of the specific goals set in our research project. Much of our work has been published. Briefly. we can summarize that: (a) Silencing of SsNox1(NADPHoxidase) expression indicated a central role for this enzyme in both virulence and pathogenic development, while inactivation of the SsNox2 gene resulted in limited sclerotial development, but the organism remained fully pathogenic. (b) A catalase gene (Scatl), whose expression was highly induced during host infection is involved in hyphal growth, branching, sclerotia formation and infection. (c) Protein tyrosine phosphatase l (ptpl) is required for sclerotial development and is involved in fungal infection. (d) Deletion of a superoxidedismutase gene (Sssodl) significantly reduced in virulence on both tomato and tobacco plants yet pathogenicity was mostly restored following supplementation with oxalate. (e) We have participated in comparative genome sequence analysis of S. sclerotiorumand B. cinerea. (f) S. sclerotiorumexhibits a potential switch between biotrophic and necrotrophic lifestyles (g) During plant­ microbe interactions cell death can occur in both resistant and susceptible events. Non­ pathogenic fungal mutants S. sclerotior111n also cause a cell death but with opposing results. We investigated PCD in more detail and showed that, although PCD occurs in both circumstances they exhibit distinctly different features. The mutants trigger a restricted cell death phenotype in the host that unexpectedly exhibits markers associated with the plant hypersensitive (resistant) response. Using electron and fluorescence microscopy, chemical effectors and reverse genetics, we have established that this restricted cell death is autophagic. Inhibition of autophagy rescued the non-pathogenic mutant phenotype. These findings indicate that autophagy is a defense response in this interaction Thus the control of cell death, dictated by the plant (autophagy) סr the fungus (apoptosis), is decisive to the outcome of certain plant­ microbe interactions. In addition to the time and efforts invested towards reaching the specific goals mentioned, both Pls have initiated utilizing (as stated as an objective in our proposal) state of the art RNA-seq tools in order to harness this technology for the study of S. sclerotiorum. The Pls have met twice (in Israel and in the US), in order to discuss .נחd coordinate the research efforts. This included a working visit at the US Pls laboratory for performing RNA-seq experiments and data analysis as well as working on a joint publication (now published). The work we have performed expands our understanding of the fundamental biology (developmental and pathogenic) of S. sclerotioז111וז. Furthermore, based on our results we have now reached the conclusion that this fungus is not a bona fide necrotroph, but can also display a biotrophic lifestyle at the early phases of infection. The data obtained can eventually serve .נ basis of rational intervention with the disease cycle of this pathogen.
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8

Droby, Samir, Michael Wisniewski, Martin Goldway, Wojciech Janisiewicz, and Charles Wilson. Enhancement of Postharvest Biocontrol Activity of the Yeast Candida oleophila by Overexpression of Lytic Enzymes. United States Department of Agriculture, November 2003. http://dx.doi.org/10.32747/2003.7586481.bard.

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Enhancing the activity of biocontrol agents could be the most important factor in their success in controlling fruit disease and their ultimate acceptance in commercial disease management. Direct manipulation of a biocontrol agent resulting in enhancement of diseases control could be achieved by using recent advances in molecular biology techniques. The objectives of this project were to isolate genes from yeast species that were used as postharvest biocontrol agents against postharvest diseases and to determine their role in biocontrol efficacy. The emphasis was to be placed on the yeast, Candida oleophila, which was jointly discovered and developed in our laboratories, and commercialized as the product, Aspire. The general plan was to develop a transformation system for C . oleophila and either knockout or overexpress particular genes of interest. Additionally, biochemical characterization of the lytic peptides was conducted in the wild-type and transgenic isolates. In addition to developing a better understanding of the mode of action of the yeast biocontrol agents, it was also our intent to demonstrate the feasibility of enhancing biocontrol activity via genetic enhancement of yeast with genes known to code for proteins with antimicrobial activity. Major achievements are: 1) Characterization of extracellular lytic enzymes produced by the yeast biocontrol agent Candida oleophila; 2) Development of a transformation system for Candida oleophila; 3) Cloning and analysis of C.oleophila glucanase gene; 4) Overexpression of and knockout of C. oleophila glucanase gene and evaluating its role in the biocontrol activity of C. oleophila; 5) Characterization of defensin gene and its expression in the yeast Pichiapastoris; 6) Cloning and Analysis of Chitinase and Adhesin Genes; 7) Characterization of the rnase secreted by C . oleophila and its inhibitory activity against P. digitatum. This project has resulted in information that enhanced our understanding of the mode of action of the yeast C . oleophila. This was important step towards enhancing the biocontrol activity of the yeast. Fungal cell wall enzymes produced by the yeast antagonist were characterized. Different substrates were identified to enhance there production in vitro. Exo-b-1, 3 glucanase, chitinase and protease production was stimulated by the presence of cell-wall fragments of Penicillium digitatum in the growing medium, in addition to glucose. A transformation system developed was used to study the role of lytic enzymes in the biocontrol activity of the yeast antagonist and was essential for genetic manipulation of C . oleqphila. After cloning and characterization of the exo-glucanase gene from the yeast, the transformation system was efficiently used to study the role of the enzyme in the biocontrol activity by over-expressing or knocking out the activity of the enzyme. At the last phase of the research (still ongoing) the transformation system is being used to study the role of chitinase gene in the mode of action. Knockout and over expression experiments are underway.
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9

Gafni, Yedidya, and Vitaly Citovsky. Inactivation of SGS3 as Molecular Basis for RNA Silencing Suppression by TYLCV V2. United States Department of Agriculture, November 2013. http://dx.doi.org/10.32747/2013.7593402.bard.

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The Israeli isolate of Tomato yellow leaf curl geminivirus(TYLCV-Is) is a major tomato pathogen, causing extensive crop losses in Israel and in the south-eastern U.S. Yet, little is known about the molecular mechanisms of its interaction with tomato cells. One of the most interesting aspects of such interaction is how the invading virus counteracts the RNA silencing response of the plant. In the former BARD project, we have shown that TYLCV-Is V2 protein is an RNA silencing suppressor, and that this suppression is carried out via the interaction of V2 with the SGS3 component of the plant RNA silencing machinery. This reported project was meant to use our data as a foundation to elucidate the molecular mechanism by which V2 affects the SGS3 activity. While this research is likely to have an important impact on our understanding of basic biology of virus-plant interactions and suppression of plant immunity, it also will have practical implications, helping to conceive novel strategies for crop resistance to TYLCV-Is. Our preliminary data in regard to V2 activities and our present knowledge of the SGS3 function suggest likely mechanisms for the inhibitory effect of V2 on SGS3. We have shown that V2 possess structural and functional hallmarks of an F-box protein, suggesting that it may target SGS3 for proteasomal degradation. SGS3 contains an RNA-binding domain and likely functions to protect the cleavage produces of the primary transcript for subsequent conversion to double-stranded forms; thus, V2 may simply block the RNA binding activity of SGS3. V2 may also employ a combination of these mechanisms. These and other possibilities were tested in this reported project.
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Bar-Joseph, Moshe, William O. Dawson, and Munir Mawassi. Role of Defective RNAs in Citrus Tristeza Virus Diseases. United States Department of Agriculture, September 2000. http://dx.doi.org/10.32747/2000.7575279.bard.

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This program focused on citrus tristeza virus (CTV), the largest and one of the most complex RNA-plant-viruses. The economic importance of this virus to the US and Israeli citrus industries, its uniqueness among RNA viruses and the possibility to tame the virus and eventually turn it into a useful tool for the protection and genetic improvement of citrus trees justify these continued efforts. Although the overall goal of this project was to study the role(s) of CTV associated defective (d)-RNAs in CTV-induced diseases, considerable research efforts had to be devoted to the engineering of the helper virus which provides the machinery to allow dRNA replication. Considerable progress was made through three main lines of complementary studies. For the first time, the generation of an engineered CTV genetic system that is capable of infecting citrus plants with in vitro modified virus was achieved. Considering that this RNA virus consists of a 20 kb genome, much larger than any other previously developed similar genetic system, completing this goal was an extremely difficult task that was accomplished by the effective collaboration and complementarity of both partners. Other full-length genomic CTV isolates were sequenced and populations examined, resulting in a new level of understanding of population complexities and dynamics in the US and Israel. In addition, this project has now considerably advanced our understanding and ability to manipulate dRNAs, a new class of genetic elements of closteroviruses, which were first found in the Israeli VT isolate and later shown to be omnipresent in CTV populations. We have characterized additional natural dRNAs and have shown that production of subgenomic mRNAs can be involved in the generation of dRNAs. We have molecularly cloned natural dRNAs and directly inoculated citrus plants with 35S-cDNA constructs and have shown that specific dRNAs are correlated with specific disease symptoms. Systems to examine dRNA replication in protoplasts were developed and the requirements for dRNA replication were defined. Several artificial dRNAs that replicate efficiently with a helper virus were created from infectious full-genomic cDNAs. Elements that allow the specific replication of dRNAs by heterologous helper viruses also were defined. The T36-derived dRNAs were replicated efficiently by a range of different wild CTV isolates and hybrid dRNAs with heterologous termini are efficiently replicated with T36 as helper. In addition we found: 1) All CTV genes except of the p6 gene product from the conserved signature block of the Closteroviridae are obligate for assembly, infectivity, and serial protoplast passage; 2) The p20 protein is a major component of the amorphous inclusion bodies of infected cells; and 3) Novel 5'-Co-terminal RNAs in CTV infected cells were characterized. These results have considerably advanced our basic understanding of the molecular biology of CTV and CTV-dRNAs and form the platform for the future manipulation of this complicated virus. As a result of these developments, the way is now open to turn constructs of this viral plant pathogen into new tools for protecting citrus against severe CTV terms and development of virus-based expression vectors for other citrus improvement needs. In conclusion, this research program has accomplished two main interconnected missions, the collection of basic information on the molecular and biological characteristics of the virus and its associated dRNAs toward development of management strategies against severe diseases caused by the virus and building of novel research tools to improve citrus varieties. Reaching these goals will allow us to advance this project to a new phase of turning the virus from a pathogen to an ally.
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