996 research outputs found

    Taxonomy of P2P Applications

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    Peer-to-peer (p2p) networks have gained immense popularity in recent years and the number of services they provide continuously rises. Where p2p-networks were formerly known as file-sharing networks, p2p is now also used for services like VoIP and IPTV. With so many different p2p applications and services the need for a taxonomy framework rises. This paper describes the available p2p applications grouped by the services they provide. A taxonomy framework is proposed to classify old and recent p2p applications based on their characteristics

    Reviewing Traffic ClassificationData Traffic Monitoring and Analysis

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    Traffic classification has received increasing attention in the last years. It aims at offering the ability to automatically recognize the application that has generated a given stream of packets from the direct and passive observation of the individual packets, or stream of packets, flowing in the network. This ability is instrumental to a number of activities that are of extreme interest to carriers, Internet service providers and network administrators in general. Indeed, traffic classification is the basic block that is required to enable any traffic management operations, from differentiating traffic pricing and treatment (e.g., policing, shaping, etc.), to security operations (e.g., firewalling, filtering, anomaly detection, etc.). Up to few years ago, almost any Internet application was using well-known transport layer protocol ports that easily allowed its identification. More recently, the number of applications using random or non-standard ports has dramatically increased (e.g. Skype, BitTorrent, VPNs, etc.). Moreover, often network applications are configured to use well-known protocol ports assigned to other applications (e.g. TCP port 80 originally reserved for Web traffic) attempting to disguise their presence. For these reasons, and for the importance of correctly classifying traffic flows, novel approaches based respectively on packet inspection, statistical and machine learning techniques, and behavioral methods have been investigated and are becoming standard practice. In this chapter, we discuss the main trend in the field of traffic classification and we describe some of the main proposals of the research community. We complete this chapter by developing two examples of behavioral classifiers: both use supervised machine learning algorithms for classifications, but each is based on different features to describe the traffic. After presenting them, we compare their performance using a large dataset, showing the benefits and drawback of each approac

    ARF GTPases and their GEFs and GAPs: concepts and challenges

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    Detailed structural, biochemical, cell biological, and genetic studies of any gene/protein are required to develop models of its actions in cells. Studying a protein family in the aggregate yields additional information, as one can include analyses of their coevolution, acquisition or loss of functionalities, structural pliability, and the emergence of shared or variations in molecular mechanisms. An even richer understanding of cell biology can be achieved through evaluating functionally linked protein families. In this review, we summarize current knowledge of three protein families: the ARF GTPases, the guanine nucleotide exchange factors (ARF GEFs) that activate them, and the GTPase-activating proteins (ARF GAPs) that have the ability to both propagate and terminate signaling. However, despite decades of scrutiny, our understanding of how these essential proteins function in cells remains fragmentary. We believe that the inherent complexity of ARF signaling and its regulation by GEFs and GAPs will require the concerted effort of many laboratories working together, ideally within a consortium to optimally pool information and resources. The collaborative study of these three functionally connected families ( \u3e /=70 mammalian genes) will yield transformative insights into regulation of cell signaling

    Fish and mammalian glut4 traffic characteristics: an evolutionary perspective on the importance of glut4 protein motifs for trafficking

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    [eng] Glucose transporters (GLUTs) are extremely important for glucose metabolism. Glucose transporters uptake glucose from blood stream into the cells where it can be metabolized. Among the glucose transporters family, GLUT4, which is solely expressed in muscle and adipose tissues, displays a unique feature as it can change its cellular distribution within minutes in response to insulin to regulate glucose uptake. Therefore, the study of GLUT4 cellular trafficking is fundamental to understand its functioning and to deepen our knowledge on glucose homeostasis. In this work, we utilized a GLUT4 fish variant, brown trout GLUT4, to study GLUT4 trafficking and the role of GLUT4 protein motifs in this process, in 3T3-L1 adipocytes. We observed that, in comparison to mammalian GLUT4 (RatGLUT4), brown trout GLUT4 (BtGLUT4) had a much weaker translocation to the plasma membrane in response to insulin which was in part due to a slower cellular trafficking (exocytosis and endocytosis) and to a poor targeting to the GLUT4 storage vesicles responsible for “holding” GLUT4 inside the cell in the absence of insulin; these vesicles represent the main pool of insulin-responsive GLUT4. In this thesis we also studied the most common GLUT4 endocytic routes. We analyzed the contribution of the clathrin-mediated and the cholesterol-dependent endocytic pathways for RatGLUT4 and BtGLUT4 internalization. We observed that whilst RatGLUT4 internalizes through both the clathrin-mediated and the cholesterol-dependent pathways, BtGLUT4 only utilizes the former. It has been suggested that in adipocytes, the main cholesterol-dependent internalization pathway is the caveolar route. The internalization through this pathway is mediated by plasma membrane structures called caveolae. The formation of these structures is dependent on the caveolin-1 protein. To analyze the role of caveolae in GLUT4 internalization we blocked its formation by knocking down caveolin-1 and observed an increase of RatGLUT4 and BtGLUT4 internalization; however, both GLUT4 isoforms showed less internalization through the clathrin-mediated and cholesterol-dependent pathways in the absence of cavolin-1. Therefore, we suggest that in 3T3-L1 adipocytes caveolin-1 knockdown induces internalization of GLUT4 through alternative pathways. GLUT4 trafficking is regulated by cellular machinery that interacts with GLUT4 protein motifs. To analyze the role of the mammalian N-terminal FQQI8 and C-terminal TELEY502 motifs in GLUT4 trafficking we mutated the corresponding motifs in BtGLUT4 (FQHL8 and TELDY495, respectively) and observed that mutations in the C-terminal had little effect on BtGLUT4 trafficking whereas mutations on the N-terminal (especially FQQL8 mutant) improved BtGLUT4 intracellular retention in the absence of insulin. Furthermore, we verified that FQQL8 mutation increased BtGLUT4 retention in a syntaxin-6-rich compartment, possibly the trans-Golgi network. In addition to studying BtGLUT4 mutants we also analyzed the trafficking of a chimera consisting of a RatGLUT4 backbone with the large cytoplasmic loop of BtGLUT4 (L-GLUT4). We observed that L-GLUT4 possessed higher plasma membrane levels in the absence of insulin and as a result a weaker translocation. Moreover, we observed that this was caused, at least in part, by a reduction in the endocytosis of L-GLUT4 in the absence of insulin. We also analyzed the contribution of the clathrin-mediated and cholesterol-dependent pathways for L-GLUT4 internalization and observed that the loop substitution (L-GLUT4) reduced RatGLUT4 internalization through the cholesterol-dependent route. Moreover, in the absence of insulin and in caveolin-1, L-GLUT4 internalization did not increase as much as that of RatGLUT4. The internalization of L-GLUT4 in the absence of caveolin-1 and insulin occurred through a clathrin-mediated pathway, similarly to BtGLUT4, but it also internalized through a cholesterol-dependent pathway, unlike RatGLUT4 and BtGLUT4. In summary, in this thesis we have contributed to increase the knowledge on GLUT4 trafficking and on the roles of the FQQI8 motif and large cytoplasmic loop in this process, in 3T3-L1 adipocytes.[spa] El transportador de glucosa GLUT4 tiene la capacidad de, en respuesta a insulina, cambiar su localizaciĂłn celular y de esta forma regular el transporte de glucosa. En este trabajo, hemos utilizado una variante de GLUT4 de trucha (BtGLUT4) para estudiar el trafico de GLUT4, asĂ­ como sus dominios proteicos involucrados en este proceso, en adipocitos 3T3-L1. Hemos observado que en comparaciĂłn con el GLUT4 de mamĂ­feros (RatGLUT4), el BtGLUT4 tenia una menor capacidad de translocaciĂłn a la membrana plasmĂĄtica en respuesta a insulina y que esto se debĂ­a a una trafico celular mas lento (exocitosis y endocitosis) y a una peor retenciĂłn en las vesĂ­culas responsables por retener el transportador dentro de la cĂ©lula en ausencia de insulina. En este trabajo hemos observado que RatGLUT4 ha internalizado por la vĂ­a de endocitosis mediada por clatrina y por la vĂ­a dependiente de colesterol, mientras que BtGLUT4 solo ha utilizado la primera. AdemĂĄs, hemos inhibido la internalizaciĂłn caveolar, mediante bajada de la expresiĂłn de caveolina-1, y hemos observado un aumento de la internalizaciĂłn de RatGLUT4 y BtGLUT4. Con el objetivo de estudiar el papel del dominio FQQI8 (extremo -N) de mamĂ­fero en el trafico de GLUT4, hemos mutado la secuencia correspondiente en BtGLUT4 (FQHL8) y hemos observado que mutaciones en este dominio han mejorado la retenciĂłn intracelular de BtGLUT4 en ausencia de insulina. TambiĂ©n hemos estudiado el trafico de una quimera que consiste en la secuencia de RatGLUT4 con el lazo citoplasmĂĄtico largo de BtGLUT4 (L-GLUT4). Hemos observado que la sustituciĂłn del lazo ha aumentado los niveles de RatGLUT4 en superficie en ausencia de insulina y que esto era debido, por lo menos en parte, a una menor endocitosis en ausencia de la hormona. TambiĂ©n hemos observado que la sustituciĂłn del lazo de RatGLUT4 ha reducido su internalizaciĂłn a travĂ©s de la vĂ­a dependiente de colesterol en ausencia de insulina. AdemĂĄs, en ausencia de caveolina-1 y insulina, la internalizaciĂłn de L-GLUT4 ha aumentado menos que la de RatGLUT4 y ha ocurrido a travĂ©s de las vĂ­as mediada por clatrina y dependiente de colesterol
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