Jaburetox (Jbtx) é um peptídeo de 10 kDa derivado de uma das isoformas de urease de Canavalia ensiformis. Em um estudo anterior realizado com o triatomíneo vetor da doença de Chagas Triatoma infestans, esse peptídeo foi encontrado interagindo com a proteína UDP-N-acetilglicosamina pirofosforilase (UAP), alterando também sua atividade enzimática no sistema nervoso central, in vivo e in vitro. A UAP já foi encontrada em eucariotos, bactérias e vírus, estando relacionada com as rotas de produção de quitina, N-glicosilação e síntese de glicoinositolfosfolipídeos. Assim, o presente trabalho tem três objetivos: i) investigar o efeito de Jbtx sobre a atividade enzimática e a expressão gênica da UAP do inseto modelo Rhodnius prolixus, ii) clonar e expressar a UAP e iii) estudar a UAP filogeneticamente. Para a primeira parte, foram avaliados, no triatomíneo R. prolixus, a atividade enzimática da UAP e o perfil de expressão dessa enzima e da quitina sintase em insetos controles e alimentados com Jbtx. Para a segunda, o cDNA da enzima de R. prolixus foi clonado em vetor pET-15b e expressado em Escherichia coli Rosetta 2. A purificação da enzima recombinante foi feita por cromatografia de afinidade a níquel. Para a terceira parte, foram buscadas sequências de aminoácidos homólogas às da UAP de R. prolixus no servidor pHmmer e foi construída uma árvore filogenética com o método de Máxima Verossimilhança. Os resultados obtidos indicam que o Jbtx aumenta a atividade enzimática da UAP em glândulas salivares, corpo gorduroso e epiderme, enquanto diminui a expressão da UAP em intestino médio anterior, túbulos de Malpighi, glândulas salivares, corpo gorduroso, epiderme e sistema nervoso central, assim como a expressão da quitina sintase nos mesmos órgãos e no intestino médio posterior. Foi obtida uma UAP recombinante de 56 kDa, compatível com peso molecular previsto in silico. A árvore filogenética construída contém 40 sequências, sendo 38 de insetos e 2 sequências de grupo externo. A árvore segue o padrão de evolução dos insetos e foi identificado um novo organismo com potenciais dois genes codificantes de UAP. Esse trabalho apresenta a primeira evidência de que Jbtx altera a expressão gênica em R. prolixus. O resultado obtido pela análise filogenética indica que a UAP é uma enzima ancestral à diversificação em Insecta. / Jaburetox (Jbtx) is a 10 kDa peptide derived from a urease isoform of Canavalia ensiformis. In a previous work with the triatomine vector of Chagas’ disease Triatoma infestans, this peptide was found interacting with the protein UDP-N-acetylglucosamine pyrophosphorylase (UAP), also increasing the UAP enzymatic activity in the central nervous system in vivo and in vitro. UAP has been described in eukaryotes, bacteria and virus, and is involved in chitin production, N-linked glycosylation and glyco inositol phospholipids synthesis pathway. Thus, the present work has three main aims: i) to understand the effect of Jbtx on this enzyme on the model insect Rhodnius prolixus, ii) to clone and express UAP and iii) to study UAP from a phylogenetic point of view. Firstly, UAP enzymatic activity and its expression profile, as well as the chitin synthase expression, were analysed in the triatomine R. prolixus in saline- or Jbtx-fed insects. Secondly, the cDNA from R. prolixus’ UAP was cloned into the pET-15b vector and expressed in Escherichia coli Rosetta 2. The recombinant enzyme was purified through a nickel affinity chromatography. Thirdly, homolog sequences to R. prolixus’ UAP were searched in pHmmer database and a phylogenetic tree was built using the Maximmum Likelihood method. The results obtained indicate that Jbtx increases UAP enzymatic activity in salivary glands, fat body and epidermis, while decreasing UAP’s expression in the anterior and posterior midgut, Malpighian tubules, salivary glands, fat body, epidermis and central nervous system, as well as the chitin synthase expression in the same organs and the posterior midgut. A 56 kDa recombinant UAP was obtained, in agreement with the in silico estimated size. The phylogenetic tree built has 40 sequences, from which 38 are from insects and 2 are from mammals (external group). The tree follows the insect evolution patterns and a new organism containing two potential UAP coding genes was identified. This work presents the first evidence that Jbtx is able to interfere in the gene expression in R. prolixus. The results obtained through phylogenetic analysis shows that UAP is an enzyme ancestral to the diversification in Insecta.

To detect network changes and network troubles, Transpondertech needs a tool that can make network measurements. The purpose of this thesis has been to find measurable network properties that best reflect the status of a network, to find methods to measure these proerties and to implement these methods in one single tool. The resulting tool is called NetworkPerf and can measure the following network properties: availability, round-trip delay, delay variation, number of hops, intermediate hosts, available bandwidth, available ports, and maximum allowed packet size. Together, these properties give a good picture of the status of a network connection. The thesis also presents the methods used for meassuring these properties in the tool.

Les récepteurs P2Y sont exprimés à la surface de bon nombre de cellules y compris les cellules du système immunitaire. De nombreuses études réalisées in vitro suggèrent une influence des nucléotides sur la biologie de ces cellules. Toutefois pour certains d’entre eux, l’implication in vivo n’est pas clairement établie. Le présent travail, basé sur l’analyse phénotypique de souris invalidées, vise donc à étudier le rôle de trois de ces récepteurs, à savoir P2Y2, P2Y6 et P2Y12, dans la physiologie des phénomènes inflammatoires. Leur rôle a été abordé plus particulièrement dans deux types cellulaires :les macrophages et les cellules dendritiques.<p>En ce qui concerne les macrophages péritonéaux recrutés, nous avons démontré in vitro que le récepteur P2Y2 participe, avec un récepteur P1, à la production d’IL-6 en réponse au LPS. Par contre, il ne semble pas impliqué dans l’internalisation d’antigènes par macropinocytose. Le récepteur P2Y6 favorise la production d’IL-6 et de MIP-2 en réponse au LPS et stimule l’endocytose par macropinocytose de dextran-FITC.<p>Pour ce qui est des cellules dendritiques (DCs), l’étude du rôle du récepteur P2Y12 dans leur biologie in vitro avait été investigué auparavant dans notre laboratoire. Toutefois, certains paramètres devaient être confirmés ainsi que l’implication in vivo du récepteur dans l’établissement d’une réponse immune. Nous avons contribué à démontrer que l’endocytose par macropinocytose est augmentée suite à l’activation du récepteur P2Y12 par l’ADPβS au niveau de DCs spléniques murines et de DCs dérivées de monocytes humains. L’ADPβS favorise la capacité des DCs à activer les lymphocytes T spécifiques d’un antigène. Enfin, la signification de ces observations a été testé in vivo par l’immunisation de souris P2Y12+/+ et P2Y12-/-. Les résultats obtenus montrent une production d’IgG1 diminuée dans les souris invalidées pour le récepteur par rapport aux souris sauvages suggérant que le récepteur P2Y12 favoriserait le développement d’une réponse immune Th2 et ce principalement via l’augmentation de la captation d’antigènes par les DCs. <p>Suite aux données obtenues et à celles de la littérature, nous avons investigué l’implication du récepteur P2Y6 dans la réponse immune adaptative in vivo. Pour ce faire, nous avons immunisé des souris P2Y6+/+ et P2Y6-/- selon le protocole utilisé lors de notre précédente étude. Les souris P2Y6 KO montrent une diminution significative de la production d’IgG2c spécifiques de l’antigène suggérant l’implication du récepteur dans le développement d’une réponse pro-Th1. Plusieurs paramètres ont été mesurés pour expliquer ce phénomène. Nous avons tout d’abord démontré que le récepteur P2Y6 est exprimé et fonctionnel dans les cellules dendritiques dérivées de la moelle. Nous avons ensuite montré que l’activation du récepteur par l’UDP favorise la capture d’antigènes in vitro, potentialise l’expression de molécules de co-stimulation (CD80 et CD86) et la production d’IL-12 p70 et de TNF-α en réponse au CpG ODN indiquant que la modulation de la réponse immune observée in vivo s’explique, en partie du moins, par les effets du récepteur sur différentes fonctions des DCs.<p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

This work is about Hierarchical agregation and it describes its parts in the rst part of this work. Next is described its parts, main features and properties in this rst part. Document describes the possibilities of realization. Second part describes an analysis of the stability problem. Realizes the detection of failures of the Feedback target stations by the implemented protocol. The protocol is programaticly realized in JAVA language and tested in the real enviroment. In this document it is described an function of bandwidth on the speed of failure detection. The issues of detection of failure on the feedback target manager machine are discussed in this document.

Abstract The current Internet infrastructure must be architected to handle future media-rich, and content rich applications. The success of applications such as video-on-demand, multicast and content distribution depends on Quality of Service and bandwidth guarantees. Over the years, the Internet has encompassed many changes in traffic profiles and applications, in bandwidths and utilization, but the future Internet infrastructure necessitates a very different architecture supporting Quality of Service (QoS). A satellite, distinguished by features such as global coverage, bandwidth flexibility, broadcast, multicast, and reliability, is an excellent candidate to provide broadband integrated Internet access. The aim of this thesis is to explore suitability of satellite technologies for broadband Internet services with significant emphasis on the question of defining, assessing, and developing QoS models for satellite ATM and IP broadband networks with and without onboard processing. For the satellite Internet, Transmission Control Protocol (TCP) performance is degraded due to long propagation delays, link errors, and bandwidth asymmetry. In this thesis, for satellite ATM, fundamental questions such as buffer requirements, TCP/ATM efficiency, fairness, and multiple access are addressed through extensive simulations in a quantitative way. Buffer designs for TCP over satellite ATM Unspecified Bit Rate (UBR) service are performed. A buffer size equal to half the round trip delay-bandwidth product of the TCP connections provides high efficiency for TCP over satellite UBR. An extensive TCP analysis via simulation study for various TCP mechanisms and end system policies show that for satellite environment end system policies are more important than switch drop policies in terms of efficiency and fairness for World Wide Web traffic. A bandwidth allocation scheme is proposed and analytical model for supporting voice and video service over a broadband satellite network is developed. The study results demonstrate that non-contiguous allocation can afford higher gain in uplink utilizations. In this thesis, for the first time, Integrated Services and Differentiated Services based QoS architectures for broadband satellite IP networks are proposed and analyzed. In multimedia applications where User Datagram Protocol (UDP) is used along with TCP, a fair excess bandwidth allocation is not possible because TCP is congestion sensitive whereas UDP is congestion insensitive. An extensive simulation model is developed to study the effect of precedence levels for reserved rate utilization and fairness with different buffer management policies. The simulation results indicate that three levels of precedence are required for better utilization. Multiprotocol Label Switching (MPLS) over Satellite network has been proposed and a simulation model developed to study the throughput performance impacts for TCP and UDP. The traffic engineering of MPLS facilitates efficient and reliable network design to optimize the utilization of network resources and enhance the network QoS. A novel Code Division Multiple Access based Spread ALOHA single code multiple access scheme for broadband satellite return channel is proposed as an alternative to Multifrequency-Time Division Multiple Access based Digital Video Broadcasting-Return Channel via Satellite protocol. It is shown through Monte Carlo simulations that throughput for Spread ALOHA One Long Code equivalent to packet length, is better than Spread ALOHA One Code in which spreading sequence repeats every symbol. The reduction of throughput due to multi-user interference for different number of users is shown. Further research on QoS architectures, performance models for TCP enhancements, interworking functions, interoperability, and standardization efforts is included.

Chlamydia trachomatis est une bactérie intracellulaire obligatoire pathogène pour l'homme, qui se développe dans un compartiment appelé inclusion. La membrane de l'inclusion constitue une protection contre les défenses de l'hôte, mais limite l'accès aux nutriments. Un élément essentiel pour C. trachomatis est le glucose. Son polymère, le glycogène, est abondant dans le lumen de l'inclusion. Ce travail a eu pour objectif de reconstituer le flux de glucose dans des cellules infectées et d'expliquer l'accumulation du glycogène. En résumé, notre travail démontre que l'accumulation de glycogène dans la lumière de l'inclusion est le résultat de deux processus, l'import de glycogène " brut " de l'hôte par invagination de la membrane de l'inclusion, et la synthèse de novo de glycogène dans le lumen de l'inclusion. Ce dernier implique l'import d'UDP-glucose par un transporteur de la cellule hôte qui est recruté dans la membrane de l'inclusion, et la sécrétion d'enzymes bactériennes dans le lumen de l'inclusion. Ces mécanismes permettent aux bactéries de stocker des molécules énergétique, inaccessibles à l'hôte. / The human pathogen Chlamydia trachomatis is an obligate intracellular bacterium, which develops in a parasitophorous compartment called inclusion. The inclusion membrane serves as a barrier to host defense mechanisms, but limits access to nutrients. One essential nutrient for C. trachomatis is glucose, and its polymer, glycogen, is highly abundant in the inclusion lumen. This work aimed to reconstitute the glucose flow in C. trachomatis infected cells and to understand the mechanisms for glycogen accumulation. In summary, our work demonstrates that glycogen storage in C. trachomatis inclusions is the result of two different strategies, bulk acquisition of host glycogen through invagination of the inclusion membrane, and de novo synthesis of glycogen within the inclusion lumen. The latter mechanism implicates the import of host UDP-glucose through a host transporter that is recruited to the inclusion membrane, and the secretion of bacterial glycogen enzymes into the inclusion lumen. These processes allow the bacteria to build an energy store within the inclusion lumen, out of reach for the host.

The aim of this diploma thesis is to create two exercises for students. The first exercise is concerned with comparison of transport protocols TCP and UDP. The second exercise is concerned with comparison of routing protocols OSPF and RIP. The first part describes used simulation environment. The next part describes the operating system used in routers. The following part describes the procedure of installing virtual machines. The next part consists of theory of protocols TCP, UDP, OSPF and RIP. The last two chapters are the laboratory exercises.

Master‘s thesis is focused on basic characteristics of data transmissions in computer networks. In forepart I digestedly described basic characteristics computer networks, metods of their measurement and valuation. Computer networks type LAN as well as wireless computer networks Wifi, are mentioned. In second part I realized my own proposition of basic characteristics measurement. This proposition is used for SW applications development. Main goal of these applications is measurement of packet delay, packet jitter, paket loss and available bandwidth. I described details of SW applications and realized measurement of real computer networks in the end.

This Graduate Thesis is about communication module for optical wireless network. This device read data from RS232 port and two analog inputs and sends it through internet to a remote Computer. Furthermore allows voice communication between local user on module and user on remote computer. Device allows communication with standard headphones and microphone, or with Bluetooth handsfree. Basic part is microprocessor MCF5223 and Bluetooth Ezurio BISM II module. Control program has been written in C language. Printed circuit is created by EAGLE program. This device is managed via web interface and settings have been stored into EEPROM memory. Work contains description of separately parts and blocks, source code of program, description of source code and printed circuit designs.

The aim of Master's thesis "Control and monitoring unit for optical link station" is design of hardware and software of unit, which provides remote control and data transfer through Internet. It is required monitoring and setting analog parameters of optical link station by web service. It includes description of Ethernet and net protocols. The core of unit are microcontroller C8051F120 and ethernet controller CP2200 made by Silicon Laboratories.