Evaluation of soybean recombinant inbred lines for seed weight yield, agronomic traits, and resistance to sudden death syndrome Sudden death syndrome (SDS) caused by Fusarium virguliforme is a devastating disease in soybean (Glycine max (L.) Merr.) that causes up to 70% of yield losses depending on the developmental stage when the plant become infected. The characterization of resistance is greatly significant for disease management. Therefore, three populations were developed by crossing three resistant lines, `Hamilton', LS90-1920 and LS97-1610 with a susceptible line to SDS, `Spencer'. Ninety-four F5:6 recombinant inbred lines from each population (Hamilton x Spencer, LS90-1920 x Spencer, and LS97-1610 x Spencer) were evaluated for two years (2009 and 2010) at two locations (Carbondale and Valmeyer) in southern Illinois. Population statistics, genotype x environment interaction, and broad-sense heritability were used to reveal any major resistance genes. Genetic correlation coefficients of SDS resistance with important agronomic traits such as lodging, pubescence, growth habit, and plant height were also calculated. The information from this study will be helpful to breeders in developing populations for genetic analyses and enforcing selection practices.

Soybean is one of the most important grain legumes grown in US and worldwide, and is a major component of human and animal protein diets. Despite improvements in management practices, and the introduction of improved soybean cultivars, soil borne pathogens continue to cause tremendous yield loss in soybean production each year. Among soil borne pathogens; Soybean Cyst Nematode (SCN) or Heterodera glycines together with Sudden Death Syndrome (SDS) induced by Fusarium virguliforme are responsible for the most damages in soybean fields. The most effective way to control these two pathogens is to develop resistant cultivars. Resistance to any population (HgType) of H. glycines, requires a functional allele at rhg1/Rfs2 locus. The rhg1/Rfs2 gene encodes a receptor-like kinase (RLK) protein. By analysing near isogenic lines (NIL) segregating for rhg1/Rfs2, rhg1-like loci were found at other locations most conservedly on LG B1. While the nature of rhg1 allele was thought to be recessive, heterozygous NIL segregating at the rhg1 locus showed that the resistant allele was dominant. Rhg1 was also inferred to be multigeneic due to absence of recombination between the RLK and other 2 genes. Functional and structural analyses were conducted on the leucine rich repeat (LRR) from RLK protein encoded by GmRLK18-1 within the Rhg1/Rfs2 locus. The LRR of GmRLK18-1 showed a high binding affinity to CLE-like peptides found in both nematode secretions and plant developmental control. Crosslinking assays and native gel analysis of GmRLK18-1-LRR validated its model as a crystal homo-dimer. Larger proteins were also shown to bind the LRR domain, in far-Western analyses both methionine synthase and cyclophilin bound strongly to the LRR domain. Homology and ab-initio modeling of the LRR domain of the GmRLK18-1 was predicted as both a monomer and a homodimer containing intrinsically unstructured regions. Amino acid substitutions found among GmRLK18-1 allotypes A87V, Q115K and H274N were predicted to play crucial roles in protein function and stability. The receptor like kinase (RLK) GmRLK18-1 within the Rhg1/Rfs2 locus underlies a pleiotropic resistance to both SCN and SDS. The resistance allele was shown to be dominant in both heterozygous NILs at Rhg1/Rfs2 and transgenics (hetero- or hemi-zygous). The RLK was found to provide a partial resistance to SCN and importantly a nearly complete resistance to both root and leaf symptoms of SDS. In the presence of Rhg4, the RLK-transgenic plants developed nearly full resistance to SCN. Therefore the RLK was proven to underlie a major portion of the Rhg1/Rfs2 locus.

Mass spectrometry is a powerful tool employed in proteomics; however, sodium dodecyl sulfate (SDS), a surfactant used for protein solubilization, is known to cause severe interference at concentrations greater than 0.01%. Thus, methods for SDS removal are paramount. This thesis presents the development of techniques for efficient SDS removal while maintaining high protein recoveries. Due to the lack of sensitivity and selectivity demonstrated by current high-throughput SDS quantitation methods, a negative-mode LC-ESI-MS technique was optimized (LOQ 0.5 ng, LOD 0.15 ng SDS). The Pierce Detergent Spin Removal Columns are a commercial product which efficiently removes SDS, but offers poor protein recovery. An alternate protocol is developed which maintains effective SDS removal while providing protein yields of >65%. Proteomic experiments often involve numerous samples, thus necessitating high-throughput methods for SDS removal. A fully automated strong cation exchange-reversed phase technique was therefore developed, which efficiently removes SDS while providing >75% protein recovery.

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Uppsatsens syfte är att ta reda på vad det finns för sociologiska aspekter som påverkar när individer röstar på Sverigedemokraterna, för att kunna ta reda på detta har jag utformat tre frågeställningar.  Jag har valt att använda mig av både sociologiska och socialpsykologiska begreppen, dessa begrepp är interaktionsritualer, grupper, jaget, identitet och stereotyper.Valet av metod baserade jag på vilken av metoderna som skulle ge det bästa svaret på mitt syfte och mina frågeställningar. Därför har jag valt att använda mig utav den kvalitativ metod då jag t.ex. kan ställa följdfrågor till intervjupersonerna och jag får en bättre inblick av hur respondenterna tycker och tänker när de svarar på frågorna och jag kan även se hur de reagerar när jag ställer frågorna och hur de använder sig av sitt kroppsspråk. Jag har valt att dela upp resultatet och analysen i fyra olika rubriker och detta gjorde jag för att det skulle vara lättare att se vad som är det viktigaste och mest intressantast i mina intervjuer men även se vad mina intervjupersoner gav för svar på mina frågeställningar. För att sedan koppla begreppen till resultatet som jag har fått från intervjuerna.   I diskussionen skriver jag om vad som hade kunnat gå bättre respektive sämre i min uppsats. För att sedan skriva om de svar jag har fått på mitt syfte och mina frågeställningar utifrån begreppen, materialet. Slutligen diskuterar jag svaren på mitt syfte och mina frågeställningar utifrån min synvinkel. / The essay's purpose is to find out the sociological aspects that influence when individuals vote for the Sweden Democrats.   I have chosen to use both sociological and social psychological concepts. These concepts are interaction rituals, groups, self, identity and stereotypes. The choice of method is based on methods that I think would give the best answer to my purpose and my questions. That is why I have chosen to use the qualitative method then for example I can put supplementary questions to the interviewees and I get a better insight of what the respondents think when replying to the questions and I can also see how they react when I ask the questions and how they use their body language.   I have chosen to divide the result and analysis in four different headings because it will be easier to see what is the most important and most interesting in my interviews, but also see the interview persons answers to my questions. Then connect the idea to the results I have received from the interviews.   In the discussion, I write about what could go better or worse in my essay. And then write about the response I have received on my purpose and my questions based on concepts and material. Last I answer to my purpose and my questions from my point of view.

This paper presents a description of the IP Multimedia Subsystem (IMS) architecture and IMS services. IMS is a standardized next-generation networking architectural framework providing advanced services on mobile and fixed networks. The first chapter describes four-layer IMS architecture and also mentions supported services. The second chapter deals with IMS protocols and primarily focuses on the SIP signaling protocol. UDP, RTP and TLS protocols are also included in this chapter. The third chapter is dedicated to practical part of this paper. A VoIP Java application has been created based on the findings gained throughout the thesis. The main part of the created application consists of client-side application, database MySQL and Servlet application for communication between database and client-side application. The whole system was created using SDS Sony Ericsson 4.1. FD1. An OpenIC lite client has been tested during development on our system and results were compared with created client-side application. A web application based on PHP, MySQL and ActionScript then handles administration and monitoring of customers using voice services.

O objetivo deste trabalho é estudar o nível de organização dos cristalitos de surfactantes aniônico, catiônico e zwiteriônico com o aumento na densidade destas moléculas sobre um cristal de germânio. As análises foram realizadas por espectroscopia de infravermelho com transformada de Fourier acoplada à técnica de reflexão total atenuada (FTIR-ATR). Este estudo apresenta importantes aspectos na organização do dodecilsulfato de sódio (SDS), do N-hexadecil-N,N-dimetil-3-amônio-1-propanosulfato (HPS), do brometo de hexadeciltrimetilamônio (CTAB) e do brometo de dodeciltrimetilamônio (DTAB). No SDS é observado um deslocamento de 1.7 cm-1 para valores de maior frequência na banda de estiramento assimétrico do CH2, vass (CH2), enquanto que é observado um deslocamento de 0.9 cm-1 na banda de estiramento simétrico, vsim (CH2). Este deslocamento para valores de maior frequência nas bandas de estiramento está associado com um aumento na desorganização da cadeia alifática com o aumento na densidade de moléculas sobre o elemento de ATR. A banda de deformação angular do CH2, δ(CH2), apresenta um valor em 1468 cm-1 que é também um indicativo de desorganização. No CTAB não é observado variações acentuadas nos valores das frequências vibracionais. Na banda vass (CH2) do DTAB é observado um deslocamento de 4.5 cm-1 para valores de menor frequência. Embora seja observado valores próximos de 2920 cm-1 para a banda vass (CH2), que é um indicativo do estado líquido de surfactantes, o que é observado nesse estudo são cristalitos de DTAB. O deslocamento da banda vsim (CH2) do DTAB é da ordem de 2 cm-1. Estas mudanças nas bandas vass (CH2) e vsim (CH2) são um indicativo da diminuição nas conformações gauche e um aumento nas conformações trans ao longo da cadeia alifática. O valor da freqüência em torno de 1472 cm-1 para a banda δ(CH2) é também um indicativo de uma maior organização na cadeia de CH2 do DTAB. Para o HPS é observado um deslocamento de 2.6 e 2.7 cm-1 para valores de maior frequência nas bandas vass (CH2) e vsim (CH2), respectivamente. A banda δ(CH2) do HPS apresenta um deslocamento de 4 cm-1 para valores de maior frequência. A variação nas bandas vass (CH2), vsim (CH2), e δ(CH2) ressalta o aumento na desorganização da cadeia alifática com o aumento na densidade de moléculas de HPS sobre o germânio. / The objective of this work is study the order level of anionic, cationic and zwitterionic surfactants with the increase of their density packing on the surface of a germanium element. The analyses were performed by a Fourier transform infrared-attenuated total reflection spectroscopy. This study shows important insights on the conformational order of sodium dodecyl sulfate (SDS), N-hexadecyl-N-N -dimethyl-3-ammonio-1-propane-sulfonate (HPS), hexadecyl-trimethylammonium bromide (CTAB) and dodecyl trimethylammonium bromide (DTAB). It is observed a shift of 1.7 and 0.9 cm-1 to higher frequency values of the CH2 asymmetric (vass (CH2)) and symmetric (vsim(CH2)) stretching bands for the SDS molecules, respectively. The latter shift to higher frequency values is associated with the disorder of the aliphatic chain due to the increase of density packing of SDS molecules on the germanium element. The CH2 scissoring band [δ (CH2)] shows a value in 1468 cm-1, which is also an indicative of conformational disorder. It is not observed any accentuated change on the vibrational frequency values of the CTAB molecules. The vass (CH2) band of the DTAB molecules is shifted 4.5 cm-1 to lower frequency values. Although it is observed values near 2920 cm-1 for the vass(CH2) band, indicating a surfactant liquid phase, it is shown in this study that there are indeed DTAB crystallites. The shift of DTAB vsim(CH2) band is around 2 cm-1. These changes in vass(CH2) and vsim(CH2) band are an indicative of a decrease in gauche conformations and an increase in all-trans conformations over the aliphatic chain. The frequency value around 1472 cm-1 for the δ(CH2) band is also an indicative of the order in CH2 chain of DTAB. It is observed a shift of 2.6 and 2.7 cm-1 to higher frequency values of vass (CH2) and vsim(CH2) bands of the HPS molecule, respectively. The δ(CH2) band of the HPS molecule presents a shift of 4 cm-1 to higher frequency values. These variations in vass (CH2), vsim(CH2), and δ(CH2) bands stand out the disorder of the aliphatic chain due to the increase of the density packing for the HPS molecules on the germanium surface.

Streptococcus pneumoniae (pneumococo) é um importante patógeno humano, responsável por graves infecções das vias respiratórias. O principal fator de virulência desse microrganismo é a cápsula polissacarídica (PS), antígeno das vacinas atuais, que são elaboradas com os PS purificados de cepas de pneumococo prevalentes na população. O objetivo desse trabalho foi desenvolver um processo de purificação do PS do sorotipo 14, responsável por 39% das doenças em crianças de 0-6 anos no Brasil. A metodologia de purificação envolveu separação celular por microfiltração tangencial e concentração do microfiltrado com membrana de ultrafiltração tangencial de 50 kDa. O produto dessa etapa foi diafiltrado com dodecil sulfato de sódio em membrana de ultrafiltração tangencial de 30 kDa, seguido de precipitação com ácido tricloroacético a 5%, precipitação por etanol (20% e 60%) e cromatografia de troca aniônica. A pureza do PS foi avaliada pelo conteúdo de proteínas e ácidos nucleicos remanescentes e o tamanho por cromatografia de exclusão molecular. O PS foi obtido com pureza e tamanho requeridos pelos órgãos regulatórios e o rendimento final do processo foi de 65%. / Streptococcus pneumoniae (pneumococcus) is an important human pathogen responsible for severe respiratory tract infections. The main virulence factor of this microorganism is the capsular polysaccharide (PS), which is the antigen of all current vaccines, which are prepared with purified PS of pneumococcal strains prevalent in the population. The objective of this work was to develop a new purification process for PS of serotype 14, responsible for 39% of diseases in children of 0-6 years old in Brazil. The purification method involved cell separation by tangential microfiltration and concentration of cell-free culture broth containing PS by tangential ultrafiltration (50 kDa). The product of this step was diafiltrated with sodium dodecyl sulfate by tangential ultrafiltration (30 kDa), following by 5% trichloroacetic acid precipitation, 20% and 60% ethanol precipitation and anion exchange chromatography. The PS purity was evaluated by the content of residual proteins and nucleic acids, and the molecular mass by size exclusion chromatography. The purity and molecular mass requirements were achieved and the process global yield was 65%.

O câncer colorretal (CCR) é a segunda de causa morte relacionada ao câncer, com aproximadamente 500.000 mortes por ano em países desenvolvidos. Os pacientes com doença localmente avançada ou metastática possuem um prognóstico ruim e, desta forma, a detecção precoce é necessária para reduzir a alta mortalidade associada à essa doença. Ainda, os métodos de rastreamento utilizados no momento apresentam boa exatidão, porém são de alto custo, consomem muito tempo e são incômodos para os pacientes. A colonoscopia é o método mais comumente utilizado para a detecção de CCR. Entretanto, apresentam duas limitações importantes: i) necessidade da luz intestinal estar limpa, sem resíduos fecais, o que só é conseguido após um preparo rigoroso, com dieta e laxativos, nos dias que antecedem a realização do exame, e, ii) é examinador-dependente, ou seja, dependem da experiência do profissional em localizar as alterações anatômicas, e ainda da habilidade em selecionar o melhor local para a realização da biópsia. Portanto, o desenvolvimento de um método rápido e sensível para o diagnóstico do câncer colorretal é extremamente importante.<br /> A proposta desta dissertação foi desenvolver um método eficaz para o preparo dos tecidos parafinizados e extração de proteínas para experimentos em eletroforese bidimensional (2DE). Tecidos fixados em formaldeído e embebidos em parafina apresentam como vantagens a preservação da qualidade morfológica, abundância e facilidade de armazenamento, o que permite estudos futuros. Entretanto, para análises proteômicas, a recuperação do material biológico é ineficiente. A partir deste estudo, 156 proteínas foram obtidas de tecidos parafinizados, sendo possível identificar por espectrometria de massas, levando a estudos posteriores para identificação de possíveis biomarcadores relacionados ao CCR. / Colorectal cancer (CRC) is the second cause of cancer-related death, with approximately 500,000 deaths per year in developed countries. Patients with locally advanced or metastatic disease have a poor prognosis and, thus, early detection is needed to reduce the high mortality associated with this disease. Yet, the screening methods used at the time have good accuracy, but are expensive, time consuming and are uncomfortable for patients. Colonoscopy is the most commonly used method for the detection of CRC. However, present two major limitations: i) the need to be clean intestinal lumen without fecal waste, which is achieved only after rigorous preparation with laxatives and diet, in the days before the exam, and ii) it is examiner-dependent, i.e., depends on the professional\'s experience in locating anatomical changes, and even the ability to select the best site for biopsy. Therefore, the development of a rapid and sensitive method for the diagnosis of colorectal cancer is extremely important. The purpose of this dissertation was to develop an effective method for the preparation of tissues and protein extraction experiments for two-dimensional electrophoresis (2DE). Tissues fixed in formaldehyde and embedded in paraffin have the advantage of preserving the morphological quality, abundance and storage facility, which allows for further studies. However, for proteomic analysis, the recovery of biological material is inefficient. From this study, 156 proteins were obtained from paraffinized tissues, and can be identified by mass spectrometry, leading to further studies to identify potential biomarkers related to CCR.

A VDAC é a proteína mais abundante da membrana mitocondrial externa. Possui diversas funções, tais como o controle da troca de metabólitos, através da membrana, e a participação no maquinário apoptótico. Estudamos o interactoma da VDAC com as proteínas mitocondriais neuronais do cérebro bovino e murino, a fim de compreender se a expressão diferenciada da VDAC1 e VDAC2 verificada entre essas células estão associadas às diferenças nas interações da VDAC. Os complexos proteicos foram analisados por 2D Blue Native SDS-PAGE e identificados via MALDI-TOF TOF usando o software Mascot e o banco de dados NCBInr. Foram identificados 27 e 46 spots em murino e bovino, respectivamente. Nós identificamos proteínas solúveis e incorporadas na membrana que não são participantes da fosforilação oxidativa, dentre elas a aldeido deidrogenase e muitas outras constituintes de complexos mitocondriais já conhecidos tão bem como novos, tais como a putative stomatin-like protein 2 complex e a switch-associated protein 70. Nossos resultados mostraram que os neurônios bovinos possuem mais complexos (5) contendo a VDAC do que em ratos (1), os quais indicam uma cinética diferencial de acoplamento e desacoplamento. Interessantemente, a lista contendo as proteínas identificadas inclui algumas proteínas conhecidas ou supostamente localizadas em compartimentos não-mitocondriais, por exemplo, a myc-induced nuclear antigen. O interactoma diferencial da VDAC entre as espécies bovina e murina, evidencia a presença de uma base comum, porém com diferentes ambientes estruturais, as quais podem ser a base da diferença entre os sítios de ligação A e B observados nas diferentes espécies. / The voltage dependent anion channel (VDAC) is the most abundant protein of outer mitochondrial membrane. VDAC controls metabolite exchange through this membrane and the apoptosis machinery. We studied the interactome of VDAC with mitochondrial proteins of neuronal cells from rat and bovine brain. We wished to understand if the differential expression of VDAC1 and VDAC2 verified between these cells was linked to differences in the VDAC interactions. Protein complexes were analyzed by 2D Blue Native SDS-PAGE and were identified by MALDI-TOF TOF using Mascot software against the NCBInr database. Number of 27 e 46 spots were identified from rat and bovine brain, respectively. We identified soluble and membrane-embedded non-OXPHOS proteins, among them aldehyde dehydrogenase, and many as constituents of known mitochondrial complexes as well as novel ones such as putative stomatin-like protein 2 complex and switchassociated protein 70. Our results showed that bovine neurons had more protein complexes (5) containing VDAC than rat cells (1), which indicates a differential kinetics of assembly or disassembly. Interestingly, the identification list included some proteins known or presumed to be localized to nonmitochondrial compartments, for example, myc-induced nuclear antigen. Our results support evidences of differential apoptotic and energetic mechanisms verified in these brains. The differential VDAC interactome between bovine and murine, support evidences of a common base, but whith different structural environment, which may be the basis of the difference between the binding sites A and B observed in these brains.