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‣ Changes in the synganglion of Rhipicephalus sanguineus (Latreille,. 1806) (Acari: Ixodidae) female ticks exposed to permethrin: An ultrastructural overview

Roma, Gislaine Cristina; Camargo Mathias, Maria Izabel; Nunes, Pablo Henrique; Bechara, Gervasio Henrique
Fonte: Elsevier B.V. Publicador: Elsevier B.V.
Tipo: Artigo de Revista Científica Formato: 19-26
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP); Processo FAPESP: 11/10427-6; Processo FAPESP: 10/51942-8; Processo FAPESP: 11/06865-8; This study performed the ultra-structural analysis of the changes caused by permethrin in the synganglion of semi-engorged Rhipicephalus sanguineus females, aiming to understand the toxic action of this substance at cellular level. The results showed that the neural lamella had its structure changed, allowing the influx of the toxic agent into the nervous tissue. The glial cells of the perineurium, as well as the neural cells of the cortex showed great changes, such as: irregular nuclei with chromatin margination, cytoplasmic vacuolation and degenerating mitochondria. These changes showed that the permethrin would be able to induce the degeneration of the synganglion through an atypical death process, involving apoptosis and autophagy. In addition, a dilated rough endoplasmic reticulum was observed in the neural cells, suggesting an intense synthesis of the hydrolytic enzymes that would be used in the processes of degradation of the damaged cellular structures (formation of lysosomes). The subperineurium and the neuropile also showed changes in their structures. Thus, it is suggested that permethrin is a dose-dependent compound able to impair the metabolism of the organism as a whole...

‣ Segregated assembly of muscle myosin expressed in nonmuscle cells.

Moncman, C L; Rindt, H; Robbins, J; Winkelmann, D A
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /10/1993 Português
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Skeletal muscle myosin cDNAs were expressed in a simian kidney cell line (COS) and a mouse myogenic cell line to investigate the mechanisms controlling early stages of myosin filament assembly. An embryonic chicken muscle myosin heavy chain (MHC) cDNA was linked to constitutive promoters from adenovirus or SV40 and transiently expressed in COS cells. These cells accumulate hybrid myosin molecules composed of muscle MHCs and endogenous, nonmuscle, myosin light chains. The muscle myosin is found associated with a Triton insoluble fraction from extracts of the COS cells by immunoprecipitation and is detected in 2.4 +/- 0.8-micron-long filamentous structures distributed throughout the cytoplasm by immunofluorescence microscopy. These structures are shown by immunoelectron microscopy to correspond to loosely organized bundles of 12-16-nm-diameter myosin filaments. The muscle and nonmuscle MHCs are segregated in the transfected cells; the endogenous nonmuscle myosin displays a normal distribution pattern along stress fibers and does not colocalize with the muscle myosin filament bundles. A similar assembly pattern and distribution are observed for expression of the muscle MHC in a myogenic cell line. The myosin assembles into filament bundles...

‣ Cellular effects of deoxynojirimycin analogues: inhibition of N-linked oligosaccharide processing and generation of free glucosylated oligosaccharides

Mellor, Howard R.; Neville, David C. A.; Harvey, David J.; Platt, Frances M.; Dwek, Raymond A.; Butters, Terry D.
Fonte: Portland Press Ltd. Publicador: Portland Press Ltd.
Tipo: Artigo de Revista Científica
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In the accompanying paper [Mellor, Neville, Harvey, Platt, Dwek and Butters (2004) Biochem. J. 381, 861–866] we treated HL60 cells with N-alk(en)yl-deoxynojirimycin (DNJ) compounds to inhibit glucosphingolipid (GSL) biosynthesis and identified a number of non-GSL-derived, small, free oligosaccharides (FOS) most likely produced due to inhibition of the oligosaccharide-processing enzymes α-glucosidases I and II. When HL60 cells were treated with concentrations of N-alk(en)ylated DNJ analogues that inhibited GSL biosynthesis completely, N-butyl- and N-nonyl-DNJ inhibited endoplasmic reticulum (ER) glucosidases I and II, but octadecyl-DNJ did not, probably due to the lack of ER lumen access for this novel, long-chain derivative. Glucosidase inhibition resulted in the appearance of free Glc1–3Man structures, which is evidence of Golgi glycoprotein endomannosidase processing of oligosaccharides with retained glucose residues. Additional large FOS was also detected in cells following a 16 h treatment with N-butyl- and N-nonyl-DNJ. When these FOS structures (>30, including >20 species not present in control cells) were characterized by enzyme digests and MALDI-TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS, all were found to be polymannose-type oligosaccharides...

‣ Real-time Imaging and Tuning Subcellular Structures and Membrane Transport Kinetics of Single Live Cells at Nanosecond Regime

Xu, Hongwu; Nallathamby, Prakash D.; Xu, Xiao-Hong Nancy
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em 29/10/2009 Português
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We developed an electric-field exposure microchannel system with 230-nanometer thin-layer gold electrodes, and interfaced it with a single living cell imaging station and a 10-nanosecond-electric-pulse (10nsEP) generator. This design allows us to image intracellular molecules and structures, membrane transport and viability of single leukemic cells (HL60) while the cells are exposed to 10nsEPs of 0–179 kV/cm, permitting the study of subcellular responses at nanosecond regime. The electrodes confine a thin-layer section of the cells exposed to 10nsEPs, offering unprecedented high spatial resolution (230-nm at z-direction of E and imaging plane) for imaging intracellular molecules of single cells affected by 10nsEPs. We found that nucleic acids, membrane transport rates and viability of single cells depend on the number and electric-field-strength (E) of 10nsEPs, showing the cumulative effect of 10nsEPs on intracellular molecules and structures and suggesting the possibility of tuning them one-pulse-at-a-time. Using lower E (51 kV/cm) of 10nsEPs, we could manipulate nucleic acids of single living cells without disrupting their cellular membrane and viability. As E increases to 80, 124 and 179 kV/cm, membrane integrity and viability of cells exhibit higher dependence on the number of 10nsEPs in a non-linear fashion...

‣ The Involvement of Lysosomes in Myocardial Aging and Disease

Terman, Alexei; Kurz, Tino; Gustafsson, Bertil; Brunk, Ulf T
Fonte: Bentham Science Publishers Ltd. Publicador: Bentham Science Publishers Ltd.
Tipo: Artigo de Revista Científica
Publicado em /05/2008 Português
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The myocardium is mainly composed of long-lived postmitotic cells with, if there is any at all, a very low rate of replacement through the division and differentiation of stem cells. As a consequence, cardiac myocytes gradually undergo pronounced age-related alterations which, furthermore, occur at a rate that inversely correlates with the longevity of species. Basically, these alterations represent the accumulation of structures that have been damaged by oxidation and that are useless and often harmful. These structures (so-called ‘waste’ materials), include defective mitochondria, aberrant cytosolic proteins, often in aggregated form, and lipofuscin, which is an intralysosomal undegradable polymeric substance. The accumulation of ‘waste’ reflects the insufficient capacity for autophagy of the lysosomal compartment, as well as the less than perfect functioning of proteasomes, calpains and other cellular digestive systems. Senescent mitochondria are usually enlarged, show reduced potential over their inner membrane, are deficient in ATP production, and often produce increased amounts of reactive oxygen species. The turnover of damaged cellular structures is hindered by an increased lipofuscin loading of the lysosomal compartment. This particularly restricts the autophagic turnover of enlarged...

‣ IRESite—a tool for the examination of viral and cellular internal ribosome entry sites

Mokrejš, Martin; Mašek, Tomáš; Vopálenský, Václav; Hlubuček, Petr; Delbos, Philippe; Pospíšek, Martin
Fonte: Oxford University Press Publicador: Oxford University Press
Tipo: Artigo de Revista Científica
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The IRESite (http://www.iresite.org) presents carefully curated experimental evidence of many eukaryotic viral and cellular internal ribosome entry site (IRES) regions. At the time of submission, IRESite stored >600 records. The IRESite gradually evolved into a robust tool providing (i) biologically meaningful information regarding the IRESs and their experimental background (including annotation of IRES secondary structures and IRES trans-acting factors) as well as (ii) thorough concluding remarks to stored database entries and regularly updated evaluation of the reported IRES function. A substantial portion of the IRESite data results purely from in-house bioinformatic analyses of currently available sequences, in silico attempts to repeat published cloning experiments, DNA sequencing and restriction endonuclease verification of received plasmid DNA. We also present a newly implemented tool for displaying RNA secondary structures and for searching through the structures currently stored in the database. The supplementary material contains an updated list of reported IRESs.

‣ Cellular pathways controlling integron cassette site folding

Loot, Céline; Bikard, David; Rachlin, Anna; Mazel, Didier
Fonte: Nature Publishing Group Publicador: Nature Publishing Group
Tipo: Artigo de Revista Científica
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By mobilizing small DNA units, integrons have a major function in the dissemination of antibiotic resistance among bacteria. The acquisition of gene cassettes occurs by recombination between the attI and attC sites catalysed by the IntI1 integron integrase. These recombination reactions use an unconventional mechanism involving a folded single-stranded attC site. We show that cellular bacterial processes delivering ssDNA, such as conjugation and replication, favour proper folding of the attC site. By developing a very sensitive in vivo assay, we also provide evidence that attC sites can recombine as cruciform structures by extrusion from double-stranded DNA. Moreover, we show an influence of DNA superhelicity on attC site extrusion in vitro and in vivo. We show that the proper folding of the attC site depends on both the propensity to form non-recombinogenic structures and the length of their variable terminal structures. These results draw the network of cell processes that regulate integron recombination.

‣ The Role of Liquid-crystalline Structures in the Morphogenesis of Animal Fibers

McKinnon, A John; Harland, Duane P
Fonte: Medknow Publications Publicador: Medknow Publications
Tipo: Artigo de Revista Científica
Publicado em //2010 Português
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The role of liquid-crystalline (mesophase) structures in extra-cellular morphogenesis is widely recognized. This paper summarizes a model for the more unusual case of intra-cellular mesophases. In the nascent mammalian hair cortex, cell differentiation is correlated with different mesophase textures within tactoids that are composed of intermediate filaments (IFs), and which form by a concerted process of unit-length-filament (ULF) polymerization and phase separation. Nematic and double-twist textures arise from differences in mesogen orientation and length in apposed tactoids. The model explains features of mature structures such as the fibril-matrix ratios in different cell types. The rapidity of IF formation suggests that a sudden-transition equilibrium polymerization, involving a high-energy initiating species, obeying the same statistical model as several other biological transitions, may be involved. This leads to an appealing symmetry, with the key factor in both polymerization and mesophase stability being the retention of protein head-group entropy.

‣ Building Complexity: Insights into Self-organized Assembly of Microtubule-based Architectures

Subramanian, Radhika; Kapoor, Tarun M.
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em 13/11/2012 Português
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Successful completion of diverse cellular functions, such as mitosis, positioning organelles, and assembling cilia, depends on the proper assembly of microtubule-based structures. While essentially all of the proteins needed to assemble these structures are now known, we cannot explain how even basic features such as size and shape are determined. As steps towards filling this knowledge gap, there have been several recent efforts towards reconstituting, with purified proteins, the basic structural motifs that recur in diverse cytoskeletal arrays. We discuss these studies and highlight how they shed light on the self-organized assembly of complex and dynamic cytoskeleton-based cellular structures.

‣ The Connectedness of Packed Circles and Spheres with Application to Conductive Cellular Materials

Swensen, John P.; Dollar, Aaron M.
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 20/12/2012 Português
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In this paper, we examine the static connectivity of 2D and 3D arrays of spherical cells with conductive paths, and the associated power dissipation in the individual cells. Herein, we use the term “cellular material” to describe the ensemble of many cells, in contrast to the more traditional use of the term for foams and honeycomb materials. Using a numerical analytical approach from highly parallel resistor arrays, we examine the cells and ensemble structures in terms of their connectivity, defined as the number of cells that are dissipating power, as well as the redundancy and robustness to localized cell failure. We examine how the connectivity changes with the geometry of the conductive cell surface area, and in particular, the percentage of the cell half that is conductive and makes contact with neighboring cells. We find that the best connectivity exists when the conductive surface of the cell is approximately 80% of the hemisphere surface, addressing the tradeoff of maximizing contact with neighboring cells while minimizing shorts in the structure. In terms of robustness, the results show that, for the proposed circular and spherical cell design, the connectivity is a nearly linear function of the number of disconnects...

‣ Rotavirus Viroplasm Proteins Interact with the Cellular SUMOylation System: Implications for Viroplasm-Like Structure Formation

Campagna, Michela; Marcos-Villar, Laura; Arnoldi, Francesca; de la Cruz-Herrera, Carlos F.; Gallego, Pedro; González-Santamaría, José; González, Dolores; Lopitz-Otsoa, Fernando; Rodriguez, Manuel S.; Burrone, Oscar R.; Rivas, Carmen
Fonte: American Society for Microbiology Publicador: American Society for Microbiology
Tipo: Artigo de Revista Científica
Publicado em /01/2013 Português
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Posttranslational modification by SUMO provides functional flexibility to target proteins. Viruses interact extensively with the cellular SUMO modification system in order to improve their replication, and there are numerous examples of viral proteins that are SUMOylated. However, thus far the relevance of SUMOylation for rotavirus replication remains unexplored. In this study, we report that SUMOylation positively regulates rotavirus replication and viral protein production. We show that SUMO can be covalently conjugated to the viroplasm proteins VP1, VP2, NSP2, VP6, and NSP5. In addition, VP1, VP2, and NSP2 can also interact with SUMO in a noncovalent manner. We observed that an NSP5 SUMOylation mutant protein retains most of its activities, such as its interaction with VP1 and NSP2, the formation of viroplasm-like structures after the coexpression with NSP2, and the ability to complement in trans the lack of NSP5 in infected cells. However, this mutant is characterized by a high degree of phosphorylation and is impaired in the formation of viroplasm-like structures when coexpressed with VP2. These results reveal for the first time a positive role for SUMO modification in rotavirus replication, describe the SUMOylation of several viroplasm resident rotavirus proteins...

‣ Actin binding proteins: Their ups and downs in metastatic life

Gross, Stephane R.
Fonte: Landes Bioscience Publicador: Landes Bioscience
Tipo: Artigo de Revista Científica
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In order to metastasize away from the primary tumor site and migrate into adjacent tissues, cancer cells will stimulate cellular motility through the regulation of their cytoskeletal structures. Through the coordinated polymerization of actin filaments, these cells will control the geometry of distinct structures, namely lamella, lamellipodia and filopodia, as well as the more recently characterized invadopodia. Because actin binding proteins play fundamental functions in regulating the dynamics of actin polymerization, they have been at the forefront of cancer research. This review focuses on a subset of actin binding proteins involved in the regulation of these cellular structures and protrusions, and presents some general principles summarizing how these proteins may remodel the structure of actin. The main body of this review aims to provide new insights into how the expression of these actin binding proteins is regulated during carcinogenesis and highlights new mechanisms that may be initiated by the metastatic cells to induce aberrant expression of such proteins.

‣ Thermodynamic perspectives on genetic instructions, the laws of biology, diseased states and human population control

Trevors, J. T.; Saier, M. H.
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
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This article examines in a broad perspective entropy and some examples of its relationship to evolution, genetic instructions and how we view diseases. Many knowledge gaps abound, hence our understanding is still fragmented and incomplete. Living organisms are programmed by functional genetic instructions (FGI), through cellular communication pathways, to grow and reproduce by maintaining a variety of hemistable, ordered structures (low entropy). Living organisms are far from equilibrium with their surrounding environmental systems, which tends towards increasing disorder (increasing entropy). Organisms must free themselves from high entropy (high disorder) to maintain their cellular structures for a period of time sufficient enough to allow reproduction and the resultant offspring to reach reproductive ages. This time interval varies for different species. Bacteria, for example need no sexual parents; dividing cells are nearly identical to the previous generation of cells, and can begin a new cell cycle without delay under appropriate conditions. By contrast, human infants require years of care before they can reproduce. Living organisms maintain order in spite of their changing surrounding environment, that decreases order according to the second law of thermodynamics. These events actually work together since living organisms create ordered biological structures by increasing local entropy. From a disease perspective...

‣ Método criptográfico baseado em autômatos celulares bidimensionais para cifragem de imagens

Magalhães Júnior, Tarcísio Abadio de
Fonte: Universidade Federal de Uberlândia Publicador: Universidade Federal de Uberlândia
Tipo: Dissertação
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Neste trabalho é proposto um novo modelo criptográfico de chave simétrica baseado em autômatos celulares bidimensionais com vizinhança von Neumann, heterogêneos e não aditivos. A cifragem do método é realizada através do cálculo de pré-imagens consecutivas e a decifragem a partir da evolução temporal para frente do autômato celular. O modelo proposto baseou-se em um trabalho anterior que utilizava autômatos celulares unidimensionais como método de cifragem, chamado Hybrid Cellular Automata (HCA). A não homogeneidade do autômato, herança do HCA, se dá pelo uso de duas regras no processo do cálculo de pré-imagens. Uma das regras é utilizada apenas nas células do contorno do reticulado a fim de garantir a existência da pré-imagem. Sua função é realizar apenas um deslocamento dos bits. A outra regra é de característica caótica e é a responsável pela cifragem efetiva do reticulado. A proposição de um novo modelo para cifrar imagens é justificada, pois os modelos convencionais de uma única dimensão não se preocupam com características espaciais das imagens. Além disso, os autômatos celulares por serem estruturas muito simples e intrinsecamente paralelos facilitam a implementação eficiente em hardware. Devido ao modelo proposto utilizar uma cifragem espacial...

‣ Origin-Independent Assembly of Kaposi's Sarcoma-Associated Herpesvirus DNA Replication Compartments in Transient Cotransfection Assays and Association with the ORF-K8 Protein and Cellular PML

Wu, Frederick Y.; Ahn, Jin-Hyun; Alcendor, Donald J.; Jang, Won-Jong; Xiao, Jinsong; Hayward, S. Diane; Hayward, Gary S.
Fonte: American Society for Microbiology Publicador: American Society for Microbiology
Tipo: Artigo de Revista Científica
Publicado em /02/2001 Português
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Six predicted Kaposi's sarcoma virus herpesvirus (KSHV) proteins have homology with other well-characterized herpesvirus core DNA replication proteins and are expected to be essential for viral DNA synthesis. Intact Flag-tagged protein products from all six were produced from genomic expression vectors, although the ORF40/41 transcript encoding a primase-helicase component proved to be spliced with a 127-bp intron. The intracellular localization of these six KSHV replication proteins and the mechanism of their nuclear translocation were investigated. SSB (single-stranded DNA binding protein, ORF6) and PPF (polymerase processivity factor, ORF59) were found to be intrinsic nuclear proteins, whereas POL (polymerase, ORF9), which localized in the cytoplasm on its own, was translocated to the nucleus when cotransfected with PPF. PAF (primase-associated factor, ORF40/41), a component of the primase-helicase tripartite subcomplex together with PRI (primase, ORF56) and HEL (helicase, ORF44), required the presence of all five other replication proteins for efficient nuclear translocation. Surprisingly, even in the absence of a lytic cycle replication origin (ori-Lyt) and any known initiator or origin binding protein, the protein products of all six KSHV core replication genes cooperated in a transient cotransfection assay to form large globular shaped pseudo-replication compartments (pseudo-RC)...

‣ Estudio de las etapas tempranas del ciclo de replicación del Virus Junín; Characterization of early steps of Junin virus replication cycle

Martínez, María Guadalupe
Fonte: Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires Publicador: Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires
Tipo: info:eu-repo/semantics/doctoralThesis; tesis doctoral; info:eu-repo/semantics/publishedVersion Formato: application/pdf
Publicado em //2010 Português
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El virus Junín (JUNV), agente etiológico de la fiebre hemorrágica argentina, es un miembro de la familia Arenaviridae. Es un virus envuelto de genoma ARN segmentado cuyo ciclo de replicación ha sido caracterizado en varios aspectos. La entrada a las células blanco se realiza vía endocitosis mediada por receptor y posterior fusión dependiente de pH. Una vez que la nucleocápside se encuentra dentro del citoplasma se expresan 5 proteínas estructurales, mediante una estrategia de codificación ambisentido. La proteína mayoritaria asociada a la nucleocápside es denominada NP. A partir de un único precursor glicoproteico (GPC), se obtienen dos glicoproteínas virales denominadas GP1 y GP2 y el péptido señal (SP). Estos conforman las estructuras claviformes encontradas en la envoltura viral. Se desconocen hasta el momento los mecanismos detallados y las estructuras celulares involucradas en los eventos tempranos del ciclo de replicación viral. En este trabajo se estudiaron aspectos de la interacción virus-célula huésped, caracterizando el rol de distintas proteínas celulares en los eventos tempranos del ciclo de multiplicación del JUNV. En primer lugar se analizó la vía de entrada utilizada por el JUNV para ser endocitado en las células...

‣ Degrees of Freedom of MIMO Cellular Networks: Decomposition and Linear Beamforming Design

Sridharan, Gokul; Yu, Wei
Fonte: Universidade Cornell Publicador: Universidade Cornell
Tipo: Artigo de Revista Científica
Publicado em 10/12/2013 Português
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This paper investigates the symmetric degrees of freedom (DoF) of MIMO cellular networks with G cells and K users per cell, having N antennas at each base station and M antennas at each user. In particular, we investigate achievability techniques based on either decomposition with asymptotic interference alignment (IA) or linear beamforming schemes, and show that there are distinct regimes of (G,K,M,N) where one outperforms the other. We first note that both one-sided and two-sided decomposition with asymptotic IA achieve the same degrees of freedom. We then establish specific antenna configurations under which the DoF achieved using decomposition based schemes is optimal by deriving a set of outer bounds on the symmetric DoF. For linear beamforming schemes, we first focus on small networks and propose a structured approach to linear beamforming based on a notion called packing ratios. Packing ratio describes the interference footprint or shadow cast by a set of transmit beamformers and enables us to identify the underlying structures for aligning interference. Such a structured beamforming design can be shown to achieve the optimal spatially normalized DoF (sDoF) of two-cell two-user/cell network and the two-cell three-user/cell network. For larger networks...

‣ Attractor structures of signaling networks: Consequences of different conformational barcode dynamics and their relations to network-based drug design

Szalay, Kristof Z.; Nussinov, Ruth; Csermely, Peter
Fonte: Universidade Cornell Publicador: Universidade Cornell
Tipo: Artigo de Revista Científica
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Conformational barcodes tag functional sites of proteins, and are decoded by interacting molecules transmitting the incoming signal. Conformational barcodes are modified by all co-occurring allosteric events induced by post-translational modifications, pathogen, drug binding, etc. We argue that fuzziness (plasticity) of conformational barcodes may be increased by disordered protein structures, by integrative plasticity of multi-phosphorylation events, by increased intracellular water content (decreased molecular crowding) and by increased action of molecular chaperones. This leads to increased plasticity of signaling and cellular networks. Increased plasticity is both substantiated by and inducing an increased noise level. Using the versatile network dynamics tool, Turbine (www.turbine.linkgroup.hu), here we show that the 10% noise level expected in cellular systems shifts a cancer-related signaling network of human cells from its proliferative attractors to its largest, apoptotic attractor representing their health-preserving response in the carcinogen containing and tumor suppressor deficient environment modeled in our study. Thus, fuzzy conformational barcodes may not only make the cellular system more plastic, and therefore more adaptable...

‣ A High-Performance Cellular Automaton Model of Tumor Growth with Dynamically Growing Domains

Poleszczuk, Jan; Enderling, Heiko
Fonte: Universidade Cornell Publicador: Universidade Cornell
Tipo: Artigo de Revista Científica
Publicado em 23/09/2013 Português
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Tumor growth from a single transformed cancer cell up to a clinically apparent mass spans many spatial and temporal orders of magnitude. Implementation of cellular automata simulations of such tumor growth can be straightforward but computing performance often counterbalances simplicity. Computationally convenient simulation times can be achieved by choosing appropriate data structures, memory and cell handling as well as domain setup. We propose a cellular automaton model of tumor growth with a domain that expands dynamically as the tumor population increases. We discuss memory access, data structures and implementation techniques that yield high-performance multi-scale Monte Carlo simulations of tumor growth. We present simulation results of the tumor growth model and discuss tumor properties that favor the proposed high-performance design.; Comment: 8 pages, 8 figures

‣ Imaging the Cell-Basement Membrane Interface during Anchor Cell Invasion in C. elegans

Hagedorn, Elliott Jennings
Fonte: Universidade Duke Publicador: Universidade Duke
Tipo: Dissertação
Publicado em //2012 Português
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Basement membrane (BM) is the thin, dense, highly cross-linked form of extracellular matrix that underlies all epithelia and endothelia, as well as surrounds muscle, nerve and fat. These sheet-like networks function as physiological barriers to maintain tissue homeostasis. During normal developmental processes and immune surveillance, cells invade through BM to establish tissues and fight infection. Similarly, metastatic cancer cells are thought to co-opt normal programs for BM transmigration as they spread from primary tumors and colonize distant tissues. The difficulty of visualizing cell-BM interactions during invasion in vivo has left the cellular and molecular mechanisms used to breach BM undefined. Specialized F-actin-rich matrix-degrading membrane protrusions, termed invadosomes, have been described in cultured invasive cell lines for more 30 years. Invadosomes are hypothesized to mediate BM penetration during cancer metastasis. Despite promising advances in intravital imaging technologies, however, invadosomes have yet to be observed in cells transmigrating BM in vivo, leaving their physiological relevance unclear. Anchor cell invasion in C. elegans is a simple in vivo model of cell invasion that allows for combined visual and genetic analysis of BM transmigration. In this dissertation I develop high-resolution time-lapse imaging approaches to understand the dynamic interactions that occur at the AC-BM interface during invasion. Through the course of this work we identify an integrin-based mechanism that polarizes the AC towards the BM. We further discover protrusive F-actin-based invadosome structures that mediate BM breach during anchor cell (AC) invasion. We find that in most cases only one or two invadosomes penetrate the BM and then transform into an invasive protrusion that guides the AC through a single BM gap. Using genetics and quantitative single-cell image analysis we characterize several molecular regulators of invadosome formation in vivo. Our findings establish an essential role for invadosomes during BM transmigration in vivo...