Bibliografías temáticas / Cell Movement

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Literatura académica sobre el tema "Cell Movement"

Autor: Grafiati
Publicado: 4 de junio de 2021
Última modificación: 6 de septiembre de 2023

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Artículos de revistas sobre el tema "Cell Movement"

1

Zigmond, Sally H. "Cell movement and cell behaviour". Cell 47, n.º 6 (diciembre de 1986): 843. http://dx.doi.org/10.1016/0092-8674(86)90798-1.

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Umetsu, Daiki, Satoshi Yamaji, Daiki Wakita y Takeshi Kano. "Quantitative Analysis of the Coordinated Movement of Cells in a Freely Moving Cell Population". Journal of Robotics and Mechatronics 35, n.º 4 (20 de agosto de 2023): 931–37. http://dx.doi.org/10.20965/jrm.2023.p0931.

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Coordinated movement of self-propelled agents has been well studied in collectives or swarms that display directional movement. Self-propelled agents also develop stable spatial patterns in which the agents do not necessarily exhibit directional collective movement. However, quantitative measures that are required to analyze the local and temporal coordinated movements during pattern formation processes have not been well established. Here, we study the coordinated movement of individual pairs of two different types of cells in a freely moving cell population. We introduced three criteria to evaluate coordinated movement in live imaging data obtained from the abdomen of the fruit fly, Drosophila melanogaster, at the pupal stage. All three criteria were able to reasonably identify coordinated movement. Our analysis indicates that the combined usage of these criteria can improve the evaluation of whether a pair of cells exhibits coordinated movement or not by excluding false positives. Quantitative approaches to identifying coordinated movement in a population of freely moving agents constitute a key foundational methodology to study pattern formations by self-propelled agents.
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LESLIE, R. J. "Molecular Motors: Cell Movement." Science 244, n.º 4912 (30 de junio de 1989): 1599. http://dx.doi.org/10.1126/science.244.4912.1599.

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Smart, Nicola y Paul R. Riley. "The Stem Cell Movement". Circulation Research 102, n.º 10 (23 de mayo de 2008): 1155–68. http://dx.doi.org/10.1161/circresaha.108.175158.

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Gough, N. R. "Caspase for Cell Movement". Science's STKE 2007, n.º 376 (28 de febrero de 2007): tw74. http://dx.doi.org/10.1126/stke.3762007tw74.

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Yoon, Jaeho, Vijay Kumar, Ravi Shankar Goutam, Sung-Chan Kim, Soochul Park, Unjoo Lee y Jaebong Kim. "Bmp Signal Gradient Modulates Convergent Cell Movement via Xarhgef3.2 during Gastrulation of Xenopus Embryos". Cells 11, n.º 1 (24 de diciembre de 2021): 44. http://dx.doi.org/10.3390/cells11010044.

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Gastrulation is a critical step in the establishment of a basic body plan during development. Convergence and extension (CE) cell movements organize germ layers during gastrulation. Noncanonical Wnt signaling has been known as major signaling that regulates CE cell movement by activating Rho and Rac. In addition, Bmp molecules are expressed in the ventral side of a developing embryo, and the ventral mesoderm region undergoes minimal CE cell movement while the dorsal mesoderm undergoes dynamic cell movements. This suggests that Bmp signal gradient may affect CE cell movement. To investigate whether Bmp signaling negatively regulates CE cell movements, we performed microarray-based screening and found that the transcription of Xenopus Arhgef3.2 (Rho guanine nucleotide exchange factor) was negatively regulated by Bmp signaling. We also showed that overexpression or knockdown of Xarhgef3.2 caused gastrulation defects. Interestingly, Xarhgef3.2 controlled gastrulation cell movements through interacting with Disheveled (Dsh2) and Dsh2-associated activator of morphogenesis 1 (Daam1). Our results suggest that Bmp gradient affects gastrulation cell movement (CE) via negative regulation of Xarhgef3.2 expression.
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Heath, Julian P. "Cell Movement and Cell Behaviour.J. M. Lackie". Quarterly Review of Biology 62, n.º 3 (septiembre de 1987): 313. http://dx.doi.org/10.1086/415542.

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McLean, Barbara Gail, Elisabeth Walgmann, Vitaly Citovsky y Patricia Zambryski. "Cell-to-cell movement of plant viruses". Trends in Microbiology 1, n.º 3 (junio de 1993): 105–9. http://dx.doi.org/10.1016/0966-842x(93)90116-9.

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Mushegian, A. R. y E. V. Koonin. "Cell-to-cell movement of plant viruses". Archives of Virology 133, n.º 3-4 (septiembre de 1993): 239–57. http://dx.doi.org/10.1007/bf01313766.

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Huckaba, Thomas M., Anna Card Gay, Luiz Fernando Pantalena, Hyeong-Cheol Yang y Liza A. Pon. "Live cell imaging of the assembly, disassembly, and actin cable–dependent movement of endosomes and actin patches in the budding yeast, Saccharomyces cerevisiae". Journal of Cell Biology 167, n.º 3 (8 de noviembre de 2004): 519–30. http://dx.doi.org/10.1083/jcb.200404173.

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Using FM4-64 to label endosomes and Abp1p-GFP or Sac6p-GFP to label actin patches, we find that (1) endosomes colocalize with actin patches as they assemble at the bud cortex; (2) endosomes colocalize with actin patches as they undergo linear, retrograde movement from buds toward mother cells; and (3) actin patches interact with and disassemble at FM4-64–labeled internal compartments. We also show that retrograde flow of actin cables mediates retrograde actin patch movement. An Arp2/3 complex mutation decreases the frequency of cortical, nonlinear actin patch movements, but has no effect on the velocity of linear, retrograde actin patch movement. Rather, linear actin patch movement occurs at the same velocity and direction as the movement of actin cables. Moreover, actin patches require actin cables for retrograde movements and colocalize with actin cables as they undergo retrograde movement. Our studies support a mechanism whereby actin cables serve as “conveyor belts” for retrograde movement and delivery of actin patches/endosomes to FM4-64–labeled internal compartments.
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Tesis sobre el tema "Cell Movement"

1

Sasaki, Nobumitsu. "Roles of Movement Protein Gene in Cell-to-Cell Movement and Host-Range Determination of Bromoviruses". Kyoto University, 2002. http://hdl.handle.net/2433/149902.

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Kyoto University (京都大学)
0048
新制・課程博士
博士(農学)
甲第9612号
農博第1240号
新制||農||841(附属図書館)
学位論文||H14||N3644(農学部図書室)
UT51-2002-G370
京都大学大学院農学研究科応用生物科学専攻
(主査)教授 奥野 哲郎, 教授 西岡 孝明, 教授 泉井 桂
学位規則第4条第1項該当
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2

Aghakhani, Minoo Razi. "Contact mediated signalling during cell movement". Thesis, King's College London (University of London), 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.404373.

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Langridge, Paul David. "Studies on cell movement using Dictyostelium". Thesis, University of Cambridge, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.613740.

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Falk, Anna. "Stem cells : proliferation, differentiation, migration /". Stockholm, 2005. http://diss.kib.ki.se/2006/91-7140-497-X/.

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Jevons, Amy Louise. "The role of PKN in cell movement". Thesis, University College London (University of London), 2008. http://discovery.ucl.ac.uk/1444205/.

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This thesis focuses on the protein kinase N family of serine/threonine protein kinases. This comprises three isoforms, with kinase domains highly related to the protein kinase C family, but with distinctive Rho/Rac dependent regulation. The critical roles played by Rho/Rac in regulating the cytoskeleton and cell migration/invasion led to an investigation into the role of PKNs in this process. Initial work in the thesis derived from the observation that PKN1 translocated to an insoluble cytosolic compartment in response to hyperosmotic stress. This had been shown to be dependent upon Racl and 3-phosphoinositide dependent kinase (PDK1). The thesis describes the characterisation of the domain in PKN 1 responsible for the Rac-dependent hyperosmotic response. Through the use of PKNl/PKCzeta chimeras a 49 amino acid sequence within the kinase domain was shown to be necessary and sufficient for the observed osmotic behaviour. In developing a cell model for migration/invasion, a breast cancer cell model was identified that displayed high PKN expression. siRNA mediated depletion of PKN isoforms or inhibition of kinase activity, revealed a requirement for PKN in both migration and invasion. Comparison with other transformed cells indicated that the relative contribution each isoform makes towards these processes appears to be cell type dependent. As a candidate downstream target, PLD has been implicated in migration and invasion of breast cancer cells and so the previously described interaction between PKN and phospholipaseD in migrating breast cancer cells was investigated. Results suggest that the activity of PLD contributes to the migration of MDAMB-468 cells and that the production of phosphatidic acid in migrating cells is stimulated by PKN.
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Atkins, David G. "Studies on the cell-to-cell movement of tobacco mosaic virus". Thesis, University of East Anglia, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.276159.

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Nagano, Hideaki. "Studies on Plant-Virus Cell-to-Cell Movement Using Chimeric Viruses". Kyoto University, 2000. http://hdl.handle.net/2433/78105.

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Kyoto University (京都大学)
0048
新制・課程博士
博士(農学)
甲第8438号
農博第1122号
新制||農||801(附属図書館)
学位論文||H12||N3395(農学部図書室)
UT51-2000-F342
京都大学大学院農学研究科農林生物学専攻
(主査)教授 古澤 巌, 教授 泉井 桂, 教授 津田 盛也
学位規則第4条第1項該当
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8

Tindall, Marcus John. "Modelling cell movement and the cell cycle in multicellular tumour spheroids". Thesis, University of Southampton, 2002. https://eprints.soton.ac.uk/50618/.

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The work presented in this thesis is concerned with modelling the effects of cell movement on the growth and formation of cell cycle phase specific regions within solid tumours. A model is proposed in the context of multicellular tumour spheroids (MCTS) and includes a simple model of the cell cycle, where cells move between each cell cycle phase depending on the availability of extracellular nutrient, as well as cell movement via chemotaxis, which varies depending upon the respective cell cycle phase of the cell. Numerical and asymptotic solutions show the model re-produces the well known MCTS structure of an internal quiescent cell region surrounded by a rim of proliferating cells. A further, more interesting result, describes a tumour surrounded by a rim of quiescent cells, with an inner quiescent and an interim proliferating cell region. The resultant solutions are a result of the different cell velocity profiles along with the effects of the cell cycle kinetics in different regions of the tumour. The non-linear form of the conservation equations describing the movement of cells means that solutions with spatial discontinuities in the cell concentrations (shocks) are observed for specific parameter values. Analysis of the effects of the chemotactic response and the cell cycle kinetics, both spatial and temporal, provide insight in to the model's behaviour and shows an understanding of cell cycle kinetics, cell movement and the spatial structure of tumours is important in assisting therapeutic strategies. The effectiveness of apoptosis, as an anti-cancer strategy, is shown to be dependent upon the concentration and spatial organisation of proliferating cells within the respective tumour. Comparison with the experimentally verified model of tumour growth developed by Gompertz allows specific model parameters to be expressed in terms of experimentally known variables. Such analysis shows that Gompertz's model is good at predicting the growth of solid tumours with a proliferating rim, but other models are required to understand the growth of non-uniform, heterogeneous tumours. Experimental justification of the model is provided by considering the observed internalisation of H3 Thymidine labelled cells and inert microspheres within MCTS. Here experimental results show that following adherence to the spheroid edge, the microspheres were all advected towards the centre of the spheroids whilst the labelled cells were spread throughout the proliferating and quiescent outer regions. The cell cycle model which is developed is, unlike previous models, able to account for this observed behaviour. Various simulations are discussed in relation to the original experimental results. These results show the importance of cell movement in providing possible ways of assisting with drug delivery to the more therapeutically resistant regions of solid tumours. Finally the importance of necrosis formation is discussed by a simple extension to the model. Necrosis as a result of quiescent cell death leads to the commonly observed formation of a necrotic core in each case. However, using the model to consider the more recent hypothesis that apoptosis leads to the formation of necrotic regions provides interesting theoretical results.
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Fernyhough, Emma Nicole. "Automated segmentation of structures essential to cell movement". Thesis, University of Leeds, 2016. http://etheses.whiterose.ac.uk/13484/.

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The study of cells is not only a key field in modern science, but has been an important area of study for hundreds of years. Despite this there is still a lot left unknown. As technology has progressed, so has our ability to photograph and film cells, but much of the processing of these images is still carried out by hand. This is not only difficult and time consuming, but is subject to opinion and error, and often not exactly reproducible. We are wishing to automate the process of segmenting cells, in order to provide biologists with that data they require to learn more about cells and their movement. This should be done in a quantitative and reproducible way. Crawling cells, such as those studied for this research, often need to move around the host body, such as the human or other mammal, in order to assist with growth, prevent disease, or to cure damage. To do this they employ other structures which protrude from the cell body to aid their motility. They use very fine hair like features (filopodia) to detect their surrounding, penetrate other cells, and determine direction. They then use thin, flat membranes (lamellipodia) to adhere both at the front and rear of the cell to pull and push forward in the direction of movement. These features are often extremely difficult to see by eye, making automation of their segmentation an awkward task. To do this, we need to use not only the information in the individual frames of video, but also information gained over time such as their movement between the frames. We firstly pre-process the images using an automated technique to correct for lighting variations in the footage. Our method is not only extremely efficient and reliable but works equally on different sizes and shapes of cell as well as frames with differing degrees of background coverage, from only one or two small cells in a frame to where the majority of the image is covered. This shading correction method was also tested on non-cellular images taken using the same kind of microscopy to show that it is suitable for all images rather than just those being studied in this work. This pre-processing allows us to make a simple segmentation of the main cell bodies, which on its own is suitable for cells which do not contain other thin structures. Using the cell bodies obtained from our pre-processing technique we then find the thinner membranes which are attached to the cell. Despite being a fully automated method, this was more accurate in two out of our three sets of videos than the most popular segmentation program using manual setting of parameters for each video individually. We improved upon this initial segmentation by incorporating the movement of the cell over time, using an iterative technique to compare the outcome of sequential frames. The result was that our segmentation was better than the manually parametrised segmentation program for every video. We then wished to find the hair like extensions and again used the information from our pre-processing stage. As these are so difficult to detect by eye we used the information of the movement to create candidate regions where these were believed to be located. Although these were usually not straight, we were able to build up small line segments in the candidate regions to recreate the features and detect the direction. This allowed us to identify all regions with filopodia present, and to separate them in order to find the required information such as the number, the length, what kind of clusters they grew in and the location compared to the direction of movement. No other method has been found which is able to detect these or segment them separately from the cell.
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Irving, Michael. "Reversible plant movement studied at single cell resolution". Thesis, Bangor University, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321434.

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Libros sobre el tema "Cell Movement"

1

Stolarska, Magdalena y Nicoleta Tarfulea, eds. Cell Movement. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1.

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1943-, Warner Fred D., Satir Peter y Gibbons Ian R, eds. Cell movement. New York: Liss, 1989.

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Cell movement and cell behaviour. London: Allen & Unwin, 1986.

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Lackie, J. M. Cell movement and cell behaviour. London: Allen & Unwin, 1986.

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Lackie, J. M. Cell Movement and Cell Behaviour. Dordrecht: Springer Netherlands, 1986. http://dx.doi.org/10.1007/978-94-009-4071-0.

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1967-, Abreu T. y Silva G. 1966-, eds. Cell movement: New research trends. Hauppauge: Nova Science Publishers, 2009.

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Bray, Dennis. Cell movements. New York: Garland Pub., 1992.

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1935-, Ishikawa Harunori, Hatano Sadashi 1929-, Satō Hidemi 1926- y Yamada Conference (10th : 1984 : Nagoya-shi, Japan), eds. Cell motility: Mechanism and regulation. New York: A.R. Liss, 1986.

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Tazawa, M. Cell Dynamics: Cytoplasmic Streaming Cell Movement-Contraction and Migration Cell and Organelle Division Phototaxis of Cell and Cell Organelle. Vienna: Springer Vienna, 1989.

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Atkins, David G. Studies on the cell-to-cell movement of tobacco mosaic virus. Norwich: University of East Anglia, 1990.

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Capítulos de libros sobre el tema "Cell Movement"

1

Lackie, J. M. "Crawling Movement". En Cell Movement and Cell Behaviour, 145–74. Dordrecht: Springer Netherlands, 1990. http://dx.doi.org/10.1007/978-94-009-4071-0_6.

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Shuttleworth, Robyn y Dumitru Trucu. "Two-Scale Moving Boundary Dynamics of Cancer Invasion: Heterotypic Cell Populations’ Evolution in Heterogeneous ECM". En Cell Movement, 1–26. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_1.

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Kim, Yangjin, Wanho Lee, Hyejin Jeon, Sookkyung Lim, Soyeon Roh, Donggu Lee, Junho Lee y Sean Lawler. "The Role of Microenvironment in Regulation of Cell Infiltration in Glioblastoma". En Cell Movement, 27–60. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_2.

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He, Xiuxiu y Yi Jiang. "A Multiscale Model of Cell Migration in Three-Dimensional Extracellular Matrix". En Cell Movement, 61–76. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_3.

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Bowman, Clark, Karen Larson, Alexander Roitershtein, Derek Stein y Anastasios Matzavinos. "Bayesian Uncertainty Quantification for Particle-Based Simulation of Lipid Bilayer Membranes". En Cell Movement, 77–102. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_4.

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Painter, Kevin J. y Thomas Hillen. "From Random Walks to Fully Anisotropic Diffusion Models for Cell and Animal Movement". En Cell Movement, 103–41. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_5.

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Xue, Chuan. "Bacterial Chemotaxis: A Classic Example of Multiscale Modeling in Biology". En Cell Movement, 143–67. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_6.

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Simons, Julie E. y Sarah D. Olson. "Sperm Motility: Models for Dynamic Behavior in Complex Environments". En Cell Movement, 169–209. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_7.

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Holz, Danielle, Laura M. McMillen, Gillian L. Ryan y Dimitrios Vavylonis. "Lamellipodia in Stationary and Fluctuating States". En Cell Movement, 211–58. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_8.

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Vasan, Ritvik, Matthew Akamatsu, Johannes Schöneberg y Padmini Rangamani. "Intracellular Membrane Trafficking: Modeling Local Movements in Cells". En Cell Movement, 259–301. Cham: Springer International Publishing, 2018. http://dx.doi.org/10.1007/978-3-319-96842-1_9.

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Actas de conferencias sobre el tema "Cell Movement"

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Nagai, Tatsuzo. "Cell movement in wounded epithelial tissues". En Third tohwa university international conference on statistical physics. AIP, 2000. http://dx.doi.org/10.1063/1.1291612.

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Hashimoto, Shigehiro. "Dielectrophoretic Movement of Cell Passing Between Surface Electrodes in Flow Channel". En ASME 2022 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2022. http://dx.doi.org/10.1115/imece2022-94776.

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Abstract In this study, cell behavior in microchannels has been tracked for the application of dielectrophoresis to biological cell selection. A pair of titanium surface electrodes integrated with a microflow channel was manufactured by photolithography technology: a triangular electrode with a tip angle of 0.26 rad and a rectangular electrode. A periodic alternating current of a square wave with a period of 0.33 μs was introduced between the electrodes to induce an asymmetric electric field perpendicular to the mainstream direction. During the flow of the suspension of mouse myoblasts (C2C12: mouse myoblast line), the behavior of the cells was measured in vitro. In addition to the shifted movement of cells by dielectrophoresis, the relationship between the distance from the electrode and the change in cell shape was investigated. Experimental results show that the distribution range of the shape of the two-dimensional projection of the cell expands under the influence of dielectrophoresis. The dielectrophoretic effect can be applied to classifying cells not only by cell size, but also by cell deformation.
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Crockett, Caroline, Elizabeth Orrico, Sara McArdle, Klaus Ley y Scott T. Acton. "Momentum measure for quantifying dendritic cell movement". En 2015 49th Asilomar Conference on Signals, Systems and Computers. IEEE, 2015. http://dx.doi.org/10.1109/acssc.2015.7421275.

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Moriizumi, Natsuki, Shigehiro Hashimoto y Shogo Uehara. "Movement of Cell Flowing Over Oblique Microgroove". En 14th International Multi-Conference on Complexity, Informatics and Cybernetics. Winter Garden, Florida, United States: International Institute of Informatics and Cybernetics, 2023. http://dx.doi.org/10.54808/imcic2023.01.17.

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Luo, Guojie. "Session details: Routing with Cell Movement (Virtual)". En ICCAD '22: IEEE/ACM International Conference on Computer-Aided Design. New York, NY, USA: ACM, 2022. http://dx.doi.org/10.1145/3578469.

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Hashimoto, Shigehiro y Hiroki Yonezawa. "Movement of Cell Flowing Over Oblique Micro Grooves in Flow Channel". En ASME 2021 Fluids Engineering Division Summer Meeting. American Society of Mechanical Engineers, 2021. http://dx.doi.org/10.1115/fedsm2021-65211.

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Abstract A micro flow-channel with bottom-microgrooves has been manufactured by photolithography technique for cell sorting. The movement of each cell passing over microgrooves has been analyzed in relation to cell deformation and alignment in vitro. The flow path (height 0.05 mm × width 1 mm × length 25 mm) between the two transparent PDMS disks has rectangular microgrooves (4.5 μm deep, 0.2 mm long) on the bottom. Variations are made in groove widths (0.03 mm, 0.04 mm, and 0.05 mm). The angle between the flow direction and the longitudinal axis of the groove is 45 degrees. Myoblasts (C2C12: mouse myoblast line) were used in the flow test. The main flow velocity of the medium (0.02 mm/s < vx < 0.23 mm/s) was controlled by the pressure difference between the inlet and the outlet. The shape of each flowing cell was tracked in a movie recorded by a camera attached to the eyepiece of the microscope. Experimental results show that the movement perpendicular to the main flow direction in the micro-groove can distinguish cells in relation to smaller deformations and larger alignment changes.
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Feher, Zoltan, Andras Veres y Zalan Heszberger. "Ping-Pong Reduction Using Sub Cell Movement Detection". En 2012 IEEE Vehicular Technology Conference (VTC 2012-Spring). IEEE, 2012. http://dx.doi.org/10.1109/vetecs.2012.6239926.

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Prieto-Langarica, A., H. V. Kojouharov, B. M. Chen-Charpentier, Michail D. Todorov y Christo I. Christov. "Mathematical Modeling of Chemoatractant Effects on Cell Movement". En APPLICATION OF MATHEMATICS IN TECHNICAL AND NATURAL SCIENCES: 3rd International Conference—AMiTaNS'11. AIP, 2011. http://dx.doi.org/10.1063/1.3659919.

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Hashimoto, Shigehiro, Taketo Matsumoto, Shogo Uehara y Yoshiaki Endo. "Bumping Movement of Cell Flowing Over Oblique Micro-Groove: Comparison with Movement Outside Groove". En 13th International Multi-Conference on Complexity, Informatics and Cybernetics. Winter Garden, Florida, United States: International Institute of Informatics and Cybernetics, 2022. http://dx.doi.org/10.54808/imcic2022.02.23.

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10

Zamir, Evan A., Brenda J. Rongish y Charles D. Little. "Dynamic Movement of Sub-Epiblastic ECM During Primitive Streak Formation". En ASME 2008 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2008. http://dx.doi.org/10.1115/sbc2008-189864.

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A well known “Polonaise” pattern of epiblast cell movements accompanies formation of the amniote primitive streak (PS), which is the organizing center for gastrulation. Although the movements observed in classical (text book) and modern studies appear similar, the biophysical mechanisms driving these movements are unknown. In comparison to studies of dynamic cellular movements during PS formation, and more generally, gastrulation, there is a relative paucity of data regarding movement of the extracellular matrix (ECM) lying adjacent to the ventral surface of the epiblast. Electron microscopy and immunofluorescence studies demonstrated decades ago the presence of a nascent basement membrane-like structure, which we refer to as the sub-epiblastic ECM (SE ECM), containing, at least, fibronectin [1] and collagen [1]. Using ultrastructural markers, Sanders [2] found that the SE ECM is transported medially to the PS with the epiblast cells. Almost two decades later, Bortier et al. [3] grafted radiolabeled quail cells into the epiblasts of chicken blastoderms, and concluded that whole groups of epiblast cells slide across (move relative to) the SE ECM — thus, contradicting Sanders’ earlier findings.
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Informes sobre el tema "Cell Movement"

1

Epel, Bernard y Roger Beachy. Mechanisms of intra- and intercellular targeting and movement of tobacco mosaic virus. United States Department of Agriculture, noviembre de 2005. http://dx.doi.org/10.32747/2005.7695874.bard.

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To cause disease, plant viruses must replicate and spread locally and systemically within the host. Cell-to-cell virus spread is mediated by virus-encoded movement proteins (MPs), which modify the structure and function of plasmodesmata (Pd), trans-wall co-axial membranous tunnels that interconnect the cytoplasm of neighboring cells. Tobacco mosaic virus (TMV) employ a single MP for cell- cell spread and for which CP is not required. The PIs, Beachy (USA) and Epel (Israel) and co-workers, developed new tools and approaches for study of the mechanism of spread of TMV that lead to a partial identification and molecular characterization of the cellular machinery involved in the trafficking process. Original research objectives: Based on our data and those of others, we proposed a working model of plant viral spread. Our model stated that MPᵀᴹⱽ, an integral ER membrane protein with its C-terminus exposed to the cytoplasm (Reichel and Beachy, 1998), alters the Pd SEL, causes the Pd cytoplasmic annulus to dilate (Wolf et al., 1989), allowing ER to glide through Pd and that this gliding is cytoskeleton mediated. The model claimed that in absence of MP, the ER in Pd (the desmotubule) is stationary, i.e. does not move through the Pd. Based on this model we designed a series of experiments to test the following questions: -Does MP potentiate ER movement through the Pd? - In the presence of MP, is there communication between adjacent cells via ER lumen? -Does MP potentiate the movement of cytoskeletal elements cell to cell? -Is MP required for cell-to-cell movement of ER membranes between cells in sink tissue? -Is the binding in situ of MP to RNA specific to vRNA sequences or is it nonspecific as measured in vitro? And if specific: -What sequences of RNA are involved in binding to MP? And finally, what host proteins are associated with MP during intracellular targeting to various subcellular targets and what if any post-translational modifications occur to MP, other than phosphorylation (Kawakami et al., 1999)? Major conclusions, solutions and achievements. A new quantitative tool was developed to measure the "coefficient of conductivity" of Pd to cytoplasmic soluble proteins. Employing this tool, we measured changes in Pd conductivity in epidermal cells of sink and source leaves of wild-type and transgenic Nicotiana benthamiana (N. benthamiana) plants expressing MPᵀᴹⱽ incubated both in dark and light and at 16 and 25 ᵒC (Liarzi and Epel, 2005 (appendix 1). To test our model we measured the effect of the presence of MP on cell-to-cell spread of a cytoplasmic fluorescent probe, of two ER intrinsic membrane protein-probes and two ER lumen protein-probes fused to GFP. The effect of a mutant virus that is incapable of cell-to-cell spread on the spread of these probes was also determined. Our data shows that MP reduces SEL for cytoplasmic molecules, dilates the desmotubule allowing cell-cell diffusion of proteins via the desmotubule lumen and reduces the rate of spread of the ER membrane probes. Replicase was shown to enhance cell-cell spread. The data are not in support of the proposed model and have led us to propose a new model for virus cell-cell spread: this model proposes that MP, an integral ER membrane protein, forms a MP:vRNAER complex and that this ER-membrane complex diffuses in the lipid milieu of the ER into the desmotubule (the ER within the Pd), and spreads cell to cell by simple diffusion in the ER/desmotubule membrane; the driving force for spread is the chemical potential gradient between an infected cell and contingent non-infected neighbors. Our data also suggests that the virus replicase has a function in altering the Pd conductivity. Transgenic plant lines that express the MP gene of the Cg tobamovirus fused to YFP under the control the ecdysone receptor and methoxyfenocide ligand were generated by the Beachy group and the expression pattern and the timing and targeting patterns were determined. A vector expressing this MPs was also developed for use by the Epel lab . The transgenic lines are being used to identify and isolate host genes that are required for cell-to-cell movement of TMV/tobamoviruses. This line is now being grown and to be employed in proteomic studies which will commence November 2005. T-DNA insertion mutagenesis is being developed to identify and isolate host genes required for cell-to-cell movement of TMV.
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2

Epel, Bernard L., Roger N. Beachy, A. Katz, G. Kotlinzky, M. Erlanger, A. Yahalom, M. Erlanger y J. Szecsi. Isolation and Characterization of Plasmodesmata Components by Association with Tobacco Mosaic Virus Movement Proteins Fused with the Green Fluorescent Protein from Aequorea victoria. United States Department of Agriculture, septiembre de 1999. http://dx.doi.org/10.32747/1999.7573996.bard.

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The coordination and regulation of growth and development in multicellular organisms is dependent, in part, on the controlled short and long-distance transport of signaling molecule: In plants, symplastic communication is provided by trans-wall co-axial membranous tunnels termed plasmodesmata (Pd). Plant viruses spread cell-to-cell by altering Pd. This movement scenario necessitates a targeting mechanism that delivers the virus to a Pd and a transport mechanism to move the virion or viral nucleic acid through the Pd channel. The identity of host proteins with which MP interacts, the mechanism of the targeting of the MP to the Pd and biochemical information on how Pd are alter are questions which have been dealt with during this BARD project. The research objectives of the two labs were to continue their biochemical, cellular and molecular studies of Pd composition and function by employing infectious modified clones of TMV in which MP is fused with GFP. We examined Pd composition, and studied the intra- and intercellular targeting mechanism of MP during the infection cycle. Most of the goals we set for ourselves were met. The Israeli PI and collaborators (Oparka et al., 1999) demonstrated that Pd permeability is under developmental control, that Pd in sink tissues indiscriminately traffic proteins of sizes of up to 50 kDa and that during the sink to source transition there is a substantial decrease in Pd permeability. It was shown that companion cells in source phloem tissue export proteins which traffic in phloem and which unload in sink tissue and move cell to cell. The TAU group employing MP:GFP as a fluorescence probe for optimized the procedure for Pd isolation. At least two proteins kinases found to be associated with Pd isolated from source leaves of N. benthamiana, one being a calcium dependent protein kinase. A number of proteins were microsequenced and identified. Polyclonal antibodies were generated against proteins in a purified Pd fraction. A T-7 phage display library was created and used to "biopan" for Pd genes using these antibodies. Selected isolates are being sequenced. The TAU group also examined whether the subcellular targeting of MP:GFP was dependent on processes that occurred only in the presence of the virus or whether targeting was a property indigenous to MP. Mutant non-functional movement proteins were also employed to study partial reactions. Subcellular targeting and movement were shown to be properties indigenous to MP and that these processes do not require other viral elements. The data also suggest post-translational modification of MP is required before the MP can move cell to cell. The USA group monitored the development of the infection and local movement of TMV in N. benthamiana, using viral constructs expressing GFP either fused to the MP of TMV or expressing GFP as a free protein. The fusion protein and/or the free GFP were expressed from either the movement protein subgenomic promoter or from the subgenomic promoter of the coat protein. Observations supported the hypothesis that expression from the cp sgp is regulated differently than expression from the mp sgp (Szecsi et al., 1999). Using immunocytochemistry and electron microscopy, it was determined that paired wall-appressed bodies behind the leading edge of the fluorescent ring induced by TMV-(mp)-MP:GFP contain MP:GFP and the viral replicase. These data suggest that viral spread may be a consequence of the replication process. Observation point out that expression of proteins from the mp sgp is temporary regulated, and degradation of the proteins occurs rapidly or more slowly, depending on protein stability. It is suggested that the MP contains an external degradation signal that contributes to rapid degradation of the protein even if expressed from the constitutive cp sgp. Experiments conducted to determine whether the degradation of GFP and MP:GFP was regulated at the protein or RNA level, indicated that regulation was at the protein level. RNA accumulation in infected protoplast was not always in correlation with protein accumulation, indicating that other mechanisms together with RNA production determine the final intensity and stability of the fluorescent proteins.
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3

Gafny, Ron, A. L. N. Rao y Edna Tanne. Etiology of the Rugose Wood Disease of Grapevine and Molecular Study of the Associated Trichoviruses. United States Department of Agriculture, septiembre de 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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4

Manulis-Sasson, Shulamit, Christine D. Smart, Isaac Barash, Laura Chalupowicz, Guido Sessa y Thomas J. Burr. Clavibacter michiganensis subsp. michiganensis-tomato interactions: expression and function of virulence factors, plant defense responses and pathogen movement. United States Department of Agriculture, febrero de 2015. http://dx.doi.org/10.32747/2015.7594405.bard.

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Clavibactermichiganensissubsp. michiganensis(Cmm), the causal agent of bacterial wilt and canker of tomato, is the most destructive bacterial disease of tomato causing substantial economic losses in Israel, the U.S.A. and worldwide. The goal of the project was to unravel the molecular strategies that allow Cmm, a Gram-positive bacterium, to develop a successful infection in tomato. The genome of Cmm contains numerous genes encoding for extracellular serine proteases and cell wall degrading enzymes. The first objective was to elucidate the role of secreted serine proteases in Cmm virulence. Mutants of nine genes encoding serine proteases of 3 different families were tested for their ability to induce wilting, when tomato stems were puncture-inoculated, as compared to blisters formation on leaves, when plants were spray-inoculated. All the mutants showed reduction in wilting and blister formation as compared to the wild type. The chpCmutant displayed the highest reduction, implicating its major role in symptom development. Five mutants of cell wall degrading enzymes and additional genes (i.e. perforin and sortase) caused wilting but were impaired in their ability to form blisters on leaves. These results suggest that Cmm differentially expressed virulence genes according to the site of penetration. Furthermore, we isolated and characterized two Cmmtranscriptional activators, Vatr1 and Vatr2 that regulate the expression of virulence factors, membrane and secreted proteins. The second objective was to determine the effect of bacterial virulence genes on movement of Cmm in tomato plants and identify the routes by which the pathogen contaminates seeds. Using a GFP-labeledCmm we could demonstrate that Cmm extensively colonizes the lumen of xylem vessels and preferentially attaches to spiral secondary wall thickening of the protoxylem and formed biofilm-like structures composed of large bacterial aggregates. Our findings suggest that virulence factors located on the chp/tomAPAI or the plasmids are required for effective movement of the pathogen in tomato and for the formation of cellular aggregates. We constructed a transposon plasmid that can be stably integrated into Cmm chromosome and express GFP, in order to follow movement to the seeds. Field strains from New York that were stably transformed with this construct, could not only access seeds systemically through the xylem, but also externally through tomato fruit lesions, which harbored high intra-and intercellular populations. Active movement and expansion of bacteria into the fruit mesocarp and nearby xylem vessels followed, once the fruit began to ripen. These results highlight the ability of Cmm to invade tomato fruit and seed through multiple entry routes. The third objective was to assess correlation between disease severity and expression levels of Cmm virulence genes and tomato defense genes. The effect of plant age on expression of tomato defense related proteins during Cmm infection was analyzed by qRT-PCR. Five genes out of eleven showed high induction at early stages of infection of plants with 19/20 leaves compared to young plants bearing 7/8 leaves. Previous results showed that Cmm virulence genes were expressed at early stages of infection in young plants compared to older plants. Results of this study suggest that Cmm virulence genes may suppress expression of tomato defense-related genes in young plants allowing effective disease development. The possibility that chpCis involved in suppression of tomato defense genes is currently under investigation by measuring the transcript level of several PR proteins, detected previously in our proteomics study. The fourth objective was to define genome location and stability of virulence genes in Cmm strains. New York isolates were compared to Israeli, Serbian, and NCPPB382 strains. The plasmid profiles of New York isolates were diverse and differed from both Israeli and Serbian strains. PCR analysis indicated that the presence of putative pathogenicity genes varied between isolates and highlighted the ephemeral nature of pathogenicity genes in field populations of Cmm. Results of this project significantly contributed to the understanding of Cmm virulence, its movement within tomato xylem or externally into the seeds, the role of serine proteases in disease development and initiated research on global regulation of Cmm virulence. These results form a basis for developing new strategies to combat wilt and canker disease of tomato.
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5

Wolf, Shmuel y William J. Lucas. Involvement of the TMV-MP in the Control of Carbon Metabolism and Partitioning in Transgenic Plants. United States Department of Agriculture, octubre de 1999. http://dx.doi.org/10.32747/1999.7570560.bard.

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The function of the 30-kilodalton movement protein (MP) of tobacco mosaic virus (TMV) is to facilitate cell-to-cell movement of viral progeny in infected plants. Our earlier findings have indicated that this protein has a direct effect on plasmodesmal function. In addition, these studies demonstrated that constitutive expression of the TMV MP gene (under the control of the CaMV 35S promoter) in transgenic tobacco plants significantly affects carbon metabolism in source leaves and alters the biomass distribution between the various plant organs. The long-term goal of the proposed research was to better understand the factors controlling carbon translocation in plants. The specific objectives were: A) To introduce into tobacco and potato plants a virally-encoded (TMV-MP) gene that affects plasmodesmal functioning and photosynthate partitioning under tissue-specific promoters. B) To introduce into tobacco and potato plants the TMV-MP gene under the control of promoters which are tightly repressed by the Tn10-encoded Tet repressor, to enable the expression of the protein by external application of tetracycline. C) To explore the mechanism by which the TMV-MP interacts with the endogenous control o~ carbon allocation. Data obtained in our previous project together with the results of this current study established that the TMV-MP has pleiotropic effects when expressed in transgenic tobacco plants. In addition to its ability to increase the plasmodesmal size exclusion limit, it alters carbohydrate metabolism in source leaves and dry matter partitioning between the various plant organs, Expression of the TMV-MP in various tissues of transgenic potato plants indicated that sugars and starch levels in source leaves are reduced below those of control plants when the TMV-MP is expressed in green tissue only. However, when the TMV-MP was expressed predominantly in PP and CC, sugar and starch levels were raised above those of control plants. Perhaps the most significant result obtained from experiments performed on transgenic potato plants was the discovery that the influence of the TMV-MP on carbohydrate allocation within source leaves was under developmental control and was exerted only during tuber development. The complexity of the mode by which the TMV-MP exerts its effect on the process of carbohydrate allocation was further demonstrated when transgenic tobacco plants were subjected to environmental stresses such as drought stress and nutrients deficiencies, Collectively, these studies indicated that the influence of the TMV-MP on carbon allocation L the result of protein-protein interaction within the source tissue. Based on these results, together with the findings that plasmodesmata potentiate the cell-to-cell trafficking of viral and endogenous proteins and nucleoproteins complexes, we developed the theme that at the whole plant level, the phloem serves as an information superhighway. Such a long-distance communication system may utilize a new class of signaling molecules (proteins and/or RNA) to co-ordinate photosynthesis and carbon/nitrogen metabolism in source leaves with the complex growth requirements of the plant under the prevailing environmental conditions. The discovery that expression of viral MP in plants can induce precise changes in carbon metabolism and photoassimilate allocation, now provide a conceptual foundation for future studies aimed at elucidating the communication network responsible for integrating photosynthetic productivity with resource allocation at the whole-plant level. Such information will surely provide an understanding of how plants coordinate the essential physiological functions performed by distantly-separated organs. Identification of the proteins involved in mediating and controlling cell-to-cell transport, especially at the companion cell-sieve element boundary, will provide an important first step towards achieving this goal.
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6

Springer, Martin, Laura Spinella, Timothy Silverman, Nick Bosco, Junki Joe, Matthew Young y Ingrid Repins. Ribbons Affect Movement of Cracked Solar Cells. Office of Scientific and Technical Information (OSTI), junio de 2023. http://dx.doi.org/10.2172/1988153.

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7

Gafni, Yedidya y Vitaly Citovsky. Molecular interactions of TYLCV capsid protein during assembly of viral particles. United States Department of Agriculture, abril de 2007. http://dx.doi.org/10.32747/2007.7587233.bard.

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Tomato yellow leaf curl geminivirus (TYLCV) is a major pathogen of cultivated tomato, causing up to 100% crop loss in many parts of the world. The present proposal, a continuation of a BARD-funded project, expanded our understanding of the molecular mechanisms by which CP molecules, as well as its pre-coat partner V2, interact with each other (CP), with the viral genome, and with cellular proteins during assembly and movement of the infectious virions. Specifically, two major objectives were proposed: I. To study in detail the molecular interactions between CP molecules and between CP and ssDNA leading to assembly of infectious TYLCV virions. II. To study the roles of host cell factors in TYLCV assembly. Our research toward these goals has produced the following major achievements: • Characterization of the CP nuclear shuttling interactor, karyopherin alpha 1, its pattern of expression and the putative involvement of auxin in regulation of its expression. (#1 in our list of publication, Mizrachy, Dabush et al. 2004). • Identify a single amino acid in the capsid protein’s sequence that is critical for normal virus life-cycle. (#2 in our list of publications, Yaakov, Levy et al. in preparation). • Development of monoclonal antibodies with high specificity to the capsid protein of TYLCV. (#3 in our list of publications, Solmensky, Zrachya et al. in press). • Generation of Tomato plants resistant to TYLCV by expressing transgene coding for siRNA targeted at the TYLCV CP. (#4 in our list of publications, Zrachya, Kumar et al. in press). •These research findings provided significant insights into (i) the molecular interactions of TYLCV capsid protein with the host cell nuclear shuttling receptor, and (ii) the mechanism by which TYLCV V2 is involved in the silencing of PTGS and contributes to the virus pathogenicity effect. Furthermore, the obtained knowledge helped us to develop specific strategies to attenuate TYLCV infection, for example, by blocking viral entry into and/or exit out of the host cell nucleus via siRNA as we showed in our publication recently (# 4 in our list of publications). Finally, in addition to the study of TYLCV nuclear import and export, our research contributed to our understanding of general mechanisms for nucleocytoplasmic shuttling of proteins and nucleic acids in plant cells. Also integration for stable transformation of ssDNA mediated by our model pathogen Agrobacterium tumefaciens led to identification of plant specific proteins involved.
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8

Sadot, Einat, Christopher Staiger y Zvi Kam Weizmann. functional genomic screen for new plant cytoskeletal proteins and the determination of their role in actin mediated functions and guard cells regulation. United States Department of Agriculture, enero de 2003. http://dx.doi.org/10.32747/2003.7587725.bard.

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The original objectives of the approved proposal were: 1. To construct a YFP fused Arabidopsis cDNA library in a mammalian expression vector. 2. To infect the library into a host fibroblast cell line and to screen for new cytoskeletal associated proteins using an automated microscope. 3. Isolate the new genes. 4. Characterize their role in plants. The project was approved as a feasibility study to allow proof of concept that would entail building the YFP library and picking up a couple of positive clones using the fluorescent screen. We report here on the construction of the YFP library, the development of the automatic microscope, the establishment of the screen and the isolation of positive clones that are plant cDNAs encoding cytoskeleton associated proteins. The rational underling a screen of plant library in fibroblasts is based on the high conservation of the cytoskeleton building blocks, actin and tubulin, between the two kingdoms (80-90% homology at the level of amino acids sequence). In addition, several publications demonstrated the recognition of mammalian cytoskeleton by plant cytoskeletal binding proteins and vice versa. The major achievements described here are: 1. The development of an automated microscope equipped with fast laser auto-focusing for high magnification and a software controlling 6 dimensions; X, Y position, auto focus, time, color, and the distribution and density of the fields acquired. This system is essential for the high throughput screen. 2. The construction of an extremely competent YFP library efficiently cloned (tens of thousands of clones collected, no empty vectors detected) with all inserts oriented 5't03'. These parameters render it well representative of the whole transcriptome and efficient in "in-frame" fusion to YFP. 3. The strategy developed for the screen allowing the isolation of individual positive cDNA clones following three rounds of microscopic scans. The major conclusion accomplished from the work described here is that the concept of using mammalian host cells for fishing new plant cytoskeletal proteins is feasible and that screening system developed is complete for addressing one of the major bottlenecks of the plant cytoskeleton field: the need for high throughput identification of functionally active cytoskeletal proteins. The new identified plant cytoskeletal proteins isolated in the pilot screen and additional new proteins which will be isolated in a comprehensive screen will shed light on cytoskeletal mediated processes playing a major role in cellular activities such as cell division, morphogenesis, and functioning such as chloroplast positioning, pollen tube and root hair elongation and the movement of guard cells. Therefore, in the long run the screen described here has clear agricultural implications.
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9

Grumet, Rebecca y Benjamin Raccah. Identification of Potyviral Domains Controlling Systemic Infection, Host Range and Aphid Transmission. United States Department of Agriculture, julio de 2000. http://dx.doi.org/10.32747/2000.7695842.bard.

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Potyviruses form one of the largest and most economically important groups of plant viruses. Individual potyviruses and their isolates vary in symptom expression, host range, and ability to overcome host resistance genes. Understanding factors influencing these biological characteristics is of agricultural importance for epidemiology and deployment of resistance strategies. Cucurbit crops are subject to severe losses by several potyviruses including the highly aggressive and variable zucchini yellow mosaic virus (ZYMV). In this project we sought to investigate protein domains in ZYMV that influence systemic infection and host range. Particular emphasis was on coat protein (CP), because of known functions in both cell to cell and long distance movement, and helper component-protease (HC-Pro), which has been implicated to play a role in symptom development and long distance movement. These two genes are also essential for aphid mediated transmission, and domains that influence disease development may also influence transmissibility. The objectives of the approved BARD project were to test roles of specific domains in the CP and HC-Pro by making sequence alterations or switches between different isolates and viruses, and testing for infectivity, host range, and aphid transmissibility. These objectives were largely achieved as described below. Finally, we also initiated new research to identify host factors interacting with potyviral proteins and demonstrated interaction between the ZYMV RNA dependent RNA polymerase and host poly-(A)-binding protein (Wang et al., in press). The focus of the CP studies (MSU) was to investigate the role of the highly variable amino terminus (NT) in host range determination and systemic infection. Hybrid ZYMV infectious clones were produced by substituting the CP-NT of ZYMV with either the CP-NT from watermelon mosaic virus (overlapping, but broader host range) or tobacco etch virus (TEV) (non- overlapping host range) (Grumet et al., 2000; Ullah ct al., in prep). Although both hybrid viruses initially established systemic infection, indicating that even the non-cucurbit adapted TEV CP-NT could facilitate long distance transport in cucurbits, after approximately 4-6, the plants inoculated with the TEV-CPNT hybrid exhibited a distinct recovery of reduced symptoms, virus titer, and virus specific protection against secondary infection. These results suggest that the plant recognizes the presence of the TEV CP-NT, which has not been adapted to infection of cucurbits, and initiates defense responses. The CP-NT also appears to play a role in naturally occurring resistance conferred by the zym locus in the cucumber line 'Dina-1'. Patterns of virus accumulation indicated that expression of resistance is developmentally controlled and is due to a block in virus movement. Switches between the core and NT domains of ZYMV-NAA (does not cause veinal chlorosis on 'Dina-1'), and ZYMV-Ct (causes veinal chlorosis), indicated that the resistance response likely involves interaction with the CP-NT (Ullah and Grumet, submitted). At the Volcani Center the main thrust was to identify domains in the HC-Pro that affect symptom expression or aphid transmissibility. From the data reported in the first and second year report and in the attached publications (Peng et al. 1998; Kadouri et al. 1998; Raccah et al. 2000: it was shown that: 1. The mutation from PTK to PAK resulted in milder symptoms of the virus on squash, 2. Two mutations, PAK and ATK, resulted in total loss of helper activity, 3. It was established for the first time that the PTK domain is involved in binding of the HC-Pro to the potyvirus particle, and 4. Some of these experiments required greater amount of HC-Pro, therefore a simpler and more efficient purification method was developed based on Ni2+ resin.
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Naim, Michael, Andrew Spielman, Shlomo Nir y Ann Noble. Bitter Taste Transduction: Cellular Pathways, Inhibition and Implications for Human Acceptance of Agricultural Food Products. United States Department of Agriculture, febrero de 2000. http://dx.doi.org/10.32747/2000.7695839.bard.

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Historically, the aversive response of humans and other mammals to bitter-taste substances has been useful for survival, since many toxic constituents taste bitter. Today, the range of foods available is more diverse. Many bitter foods are not only safe for consumption but contain bitter constituents that provide nutritional benefits. Despite this, these foods are often eliminated from our current diets because of their unacceptable bitterness. Extensive technology has been developed to remove or mask bitterness in foods, but a lack of understanding of the mechanisms of bitterness perception at the taste receptor level has prevented the development of inhibitors or efficient methods for reducing bitterness. In our original application we proposed to: (a) investigate the time course and effect of selected bitter tastants relevant to agricultural products on the formation of intracellular signal molecules (cAMP, IP3, Ca2+) in intact taste cells, in model cells and in membranes derived therefrom; (b) study the effect of specific bitter taste inhibitors on messenger formation and identify G-proteins that may be involved in tastant-induced bitter sensation; (c) investigate interactions and self-aggregation of bitter tastants within membranes; (d) study human sensory responses over time to these bitter-taste stimuli and inhibitors in order to validate the biochemical data. Quench-flow module (QFM) and fast pipetting system (FPS) allowed us to monitor fast release of the aforementioned signal molecules (cGMP, as a putative initial signal was substituted for Ca2+ ions) - using taste membranes and intact taste cells in a time range below 500 ms (real time of taste sensation) - in response to bitter-taste stimulation. Limonin (citrus) and catechin (wine) were found to reduce cellular cAMP and increase IP3 contents. Naringin (citrus) stimulated an IP3 increase whereas the cheese-derived bitter peptide cyclo(leu-Trp) reduced IP3 but significantly increased cAMP levels. Thus, specific transduction pathways were identified, the results support the notion of multiple transduction pathways for bitter taste and cross-talk between a few of those transduction pathways. Furthermore, amphipathic tastants permeate rapidly (within seconds) into liposomes and taste cells suggesting their availability for direct activation of signal transduction components by means of receptor-independent mechanisms within the time course of taste sensation. The activation of pigment movement and transduction pathways in frog melanophores by these tastants supports such mechanisms. Some bitter tastants, due to their amphipathic properties, permeated (or interacted with) into a bitter tastant inhibitor (specific phospholipid mixture) which apparently forms micelles. Thus, a mechanism via which this bitter taste inhibitor acts is proposed. Human sensory evaluation experiments humans performed according to their 6-n-propyl thiouracil (PROP) status (non-tasters, tasters, super-tasters), indicated differential perception of bitterness threshold and intensity of these bitter compounds by different individuals independent of PROP status. This suggests that natural products containing bitter compounds (e.g., naringin and limonin in citrus), are perceived very differently, and are in line with multiple transduction pathways suggested in the biochemical experiments. This project provides the first comprehensive effort to explore the molecular basis of bitter taste at the taste-cell level induced by economically important and agriculturally relevant food products. The findings, proposing a mechanism for bitter-taste inhibition by a bitter taste inhibitor (made up of food components) pave the way for the development of new, and perhaps more potent bitter-taste inhibitors which may eventually become economically relevant.
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