Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2018-03-26T14:34:13Z No. of bitstreams: 1 2015 - Fernando de Azevedo da Cruz Seara.pdf: 1716878 bytes, checksum: 6a6ea74b23b441a8aafb15e937dea036 (MD5) / Made available in DSpace on 2018-03-26T14:34:14Z (GMT). No. of bitstreams: 1 2015 - Fernando de Azevedo da Cruz Seara.pdf: 1716878 bytes, checksum: 6a6ea74b23b441a8aafb15e937dea036 (MD5) Previous issue date: 2015-07-24 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / According to World Health Organization, ischemia heart diseases are the leading cause of death worldwide. Among therapeutic approaches, reperfusion is the most effective and indicated is reperfusion. Despite the better post-infarction prognostic, absolute improvement on cardiac function is hardly achieved due to ischemia/reperfusion injury (IRI). Within this context, anabolic steroids (AS) administration, in adult Wistar rats, significantly increase IRI susceptibility. Moreover, chronic administration of AS, during adolescent phase, induces persistent cardiovascular dysfunctions along adulthood. Therefore, the aim of the present study was to analyze the effects of chronic administration of supraphysiologic doses of testosterone propionate, during adolescent phase, in the susceptibility to ischemia/reperfusion injury, in adult Wistar rats. To perform it, 24 Wistar rats were allocated into two groups, AS (Testosterone propionate 5 mg kg-1, since 26? day postnatal, 5 days per week, during 5 weeks) and Control (Vehicle). In the 82? postnatal, rats were euthanized and hearts, livers, lungs, kidneys and testicles were collected. Isolated hearts were artificially perfused with modified Krebs-Henseleit solution, through Langendorff apparatus, and, then, submitted to ex vivo ischemia ? reperfusion protocol (20 minutes of stabilization, 30 minutes of global ischemia and 60 minutes of reperfusion). The left ventricle (LV) end diastolic- LVEDP), systolic- (LVSP) and developed pressures (LVDP), as well as first derivatives of pressure, maximum and minimum (dP/dt, maximum and minimum, respectively) were measured through an intraventricular latex balloon, connected to a pressure transducer. Through the electrocardiogram, susceptibility to arrhythmic episodes was analyzed. At the end of the protocol, area of infarct was delimited and gene expression of ? and ? myosin heavy chains and Glyceraldehyde 3-phosphate dehydrogenase, as well as the activity of nicotinamide adenine dinucleotide phosphate-oxidase (Nox) enzymes, were calculated. In comparison to Control group, hearts from AS group presented: Hypertrophy, due to an increase in cardiac mass (33%, P<0,001) and index (37%, P<0,001); Significantly increase in the area of infarct (54,76%, P<0,05); Worst recovery of both LVEDP and LVDP, along reperfusion; Less recovery of maximum dP/dt, during reperfusion, despite the equivalent LVSP; Reduced basal minimum dP/dt and, subsequently, reduction in the recovery of the aforementioned parameter, regarding reperfusion period; Enhanced gene expression of MHC? (%), consistent with the loss of mechanical performance; Increased incidence of arrhythmic episodes in the reperfusion period (100%, P<0,01). No statistical difference could be seen in regard to the Nox activity. For the first time, we demonstrated that AS treatment during adolescent phase promotes cardiac hypertrophy and gene reprogramming, both persistent during adulthood, besides an increase susceptibility to IRI, through in the larger area of infarct and poor recovery of cardiac electrical and mechanical proprieties, in isolated hearts of adult Wistar rats / De acordo com a OMS, as doen?as isqu?micas do cora??o consistem na maior causa mortis global. Dentre as abordagens terap?uticas, a mais eficaz ? a reperfus?o. A despeito da melhora no progn?stico p?s-infarto, a recupera??o plena da fun??o card?aca dificilmente ? alcan?ada, devido a inj?ria de isquemia/reperfus?o (IIR). Neste contexto, a administra??o de EA em ratos Wistar adultos enaltece a susceptibilidade ? IIR. Ademais, a administra??o de EA em ratos Wistar, ao longo da adolesc?ncia, favorece o desenvolvimento de disfun??es cardiovasculares persistentes durante a fase adulta. Desta forma, objetivou-se, com o presente estudo, analisar os efeitos da sobrecarga cr?nica de propionato de testosterona, ao longo da fase adolescente, na susceptibilidade ? IIR, em ratos Wistar adultos. Para tanto, foram utilizados 24 ratos Wistar machos, divididos em dois grupos: EA (Propionato de testosterona 5 mg kg-1, a partir do 26? dia p?s-natal, 5 vezes por semana/ 5 semanas) e CTL (ve?culo). No 82? dia p?s-natal, os ratos foram submetidos ? eutan?sia para a coleta ?rg?os. Os cora??es isolados foram submetidos ? perfus?o artificial em aparato de Langendorff, e, assim, ao protocolo de isquemia/reperfus?o. As press?es diast?lica final (PDF), sist?lica (PS) e desenvolvida (PD), do ventr?culo esquerdo (VE), e as primeiras derivadas de press?o do VE, m?xima e m?nima (dP/dt m?xima e m?nima, respectivamente), foram mensuradas atrav?s de um bal?o de l?tex intraventricular, conectado a um transdutor de press?o. Atrav?s do eletrocardiograma, foi analisada a susceptibilidade aos epis?dios arr?tmicos. Ao final do protocolo, as ?reas de infarto foram demarcadas e a express?o g?nica das cadeias pesadas de miosina e gliceralde?do-3-fosfato desidrogenase, assim como a atividade enzimas da fam?lia de niconinam?da adenina dinucleot?deo fosfato oxidase (Nox), no ventr?culo esquerdo, foram avaliadas. Em rela??o ao grupo Controle, os cora??es dos animais tratados com EA apresentaram: Hipertrofia, atrav?s do aumento na massa (aumento de 33%, P<0,001) e do ?ndice card?aco (aumento de 37%, P<0,001); aumento significativo da ?rea de infarto (aumento de 54,76%, P<0,05); T?nue recupera??o da PDFVE, assim como da PDVE, durante a reperfus?o; Inferior recupera??o da dP/dt m?xima, ao longo da reperfus?o, a despeito da equivalente recupera??o na PSVE; Reduzida dP/dt m?nima basal e, subsequentemente, redu??o na recupera??o deste par?metro, ao longo da reperfus?o; Aumento significativo da express?o g?nica da MHC? (P<0,01), condizente o preju?zo no desempenho mec?nico; Maior incid?ncia de epis?dios arr?tmicos, ao longo da reperfus?o (aumento de 100%, P<0,01). N?o houve diferen?a em rela??o ?s atividades das Nox. Pela primeira vez, foi demonstrado que a administra??o de EA, ao longo da adolesc?ncia, provoca hipertrofia e reprograma??o g?nica card?aca, persistente durante a fase adulta, al?m de aumentar, significativamente, a susceptibilidade ? IIR, por meio do aumento na ?rea de infarto e piora na recupera??o das propriedades mec?nicas e el?tricas card?acas, em cora??es isolados de ratos Wistar adultos.

Made available in DSpace on 2014-12-17T14:01:58Z (GMT). No. of bitstreams: 1 DinaelzaCP.pdf: 982597 bytes, checksum: 8505a456042640660a4eeec39570774f (MD5) Previous issue date: 2009-03-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The seaweed Gracilaria domingensis is a common species in the coast of Rio Grande do Norte. This species lives in the intertidal zone, where colour strains (red, green and brown) co-occur during the whole year. Seaweeds that live in this region are exposed to daily changes and to the rhythm of the tide. During the low tide they are exposed to dissection, hiper-or hipo-osmotic shock, high temperatures and high irradiance. The aim of this study was to analyze whether the pigment and protein content of the colour strains of G. domingensis is affected by some environmental parameters in a temporal scale. The seaweeds were collected during 10 months in the seashore of Rio do Fogo (RN). The total soluble proteins and the phycobiliprotein were extracted in phosphate buffer and the carotenoids were analyzed by a standardized method through HPLC-UV. The pigments analysis showed that phycoerithrin is the most abundant pigment in the three strains. This pigment was strongly correlated with nitrogen and the photosynthetically active radiation. Chlorophyll presented higher concentrations than carotenoids during the whole, but the ratio carotenoid/chlorophyll-a was modified by incident radiation. The most abundant carotenoid was ?-carotene and zeaxanthin, which had higher concentrations in the higher radiation months. The concentration increase of zeaxanthin in this period indicated a photoprotective response of the seaweed. The three strains presented a pigment profile that indicates different radiation tolerance profile. Our results pointed that the green strain is better adapted to high irradiance levels than the red and brown strains / A macroalga Gracilaria domingensis ? comumente encontrada no litoral do Rio Grande do Norte. Esta esp?cie habita a zona intertidal, onde linhagens crom?ticas (vermelha, verde e marrom) co-ocorrem durante todo o ano. Algas que vivem nesta regi?o est?o submetidas a mudan?as diurnas e ao ritmo da mar?. Durante a mar? baixa elas ficam expostas a disseca??o, choque hiper ou hipo-osm?tico, altas temperaturas e elevada irradi?ncia. Este trabalho teve como objetivo geral o estudo do perfil prot?ico e pigmentar das variantes crom?ticas de G. domingensis e a altera??o deste perfil em fun??o dos par?metros ambientais em uma escala temporal. As algas foram coletadas mensalmente, durante 10 meses, nas parias de Rio do Fogo-RN. As prote?nas totais sol?veis e as ficobiliprote?nas foram extra?das em tamp?o fosfato e os caroten?ides foram analisados em um m?todo padronizado por HPLC-UV. A an?lise dos pigmentos mostrou que a ficoeritrina ? o pigmento mais abundante nas tr?s linhagens. Este pigmento esteve fortemente correlacionado com o nitrog?nio e a PAR. Os pigmentos caroten?ides apresentaram concentra??es inferiores as da clorofila-a durante todos os meses, mas a raz?o caroten?ides/clorofila-a foi modificada com o aumento da radia??o. O caroten?ide mais abundante foi o ?-caroteno, seguido da zeaxantina, que esteve em maiores concentra??es nos meses de maior radia??o. O aumento na concentra??o da zeaxantina nesse per?odo indicou uma resposta fotoprotetora da alga. As tr?s linhagens apresentaram um perfil pigmentar que remete a diferentes padr?es de toler?ncia a radia??o. A linhagem verde mostrou ser melhor adaptada a elevados n?veis de irradi?ncias do que a vermelha e a marrom

Made available in DSpace on 2014-12-17T14:16:30Z (GMT). No. of bitstreams: 1 AnneCOC_DISSERT.pdf: 1977864 bytes, checksum: 512fa415d2a3e86643a5d0929e22bccf (MD5) Previous issue date: 2012-06-05 / Kalanchoe brasiliensis Cambess (Crassulaceae), commonly known as sai?o , coirama branca , folha grossa , is originally from Brazil and commonly found in S?o Paulo to Bahia, mainly in the coastal zone. Regarding of biological activities, most preclinical studies were found in the literature, mainly about the anti-inflammatory activity of extracts obtained from leaves and / or aerial parts of K. brasiliensis. As regards the chemical constitution, it has been reported mainly the presence of flavonoids in the leaves of the species, but until this moment did not knows which are the active compounds. Although it is a species widely used in traditional medicine in Brazil, there is no monograph about the quality parameters of the plant drug. In this context, this study aims to characterize and quantify the chemical markers of hydroethanolic extract (HE) from the leaves of K. brasiliensis, which can be used in quality control of plant drug and derivatives obtained from this species. The methodology was divided into two parts: i. Phytochemical study: to fractionate, isolate and characterizate of the chemical (s) marker (s) of the HE from the leaves of K. brasiliensis; ii. To Developed validate of analytical method by High Performance Liquid Chromatography (HPLC)-diode array detector (DAD) to quantify the chemical (s) marker (s) of the EH. i. The EH 50% was prepared by turbo extraction method. It was then submitted to liquid-liquid partition, obtaining dichloromethane, n-butanol and ethyl acetate (AcOEt) fractions. The AcOEt fraction was selected to continue the fractionation process, because it has a chemical profile rich in flavonoids. The acOEt fraction was submitted to column chromatography using different systems for obtaining the compound Kb1. To identify this compound, it was submitted to UV analysis ii. For quantitative analysis, the EH was analyzed by HPLC, using different methods. After selecting the most appropriate method, which showed satisfactory resolution and symmetrical peaks, it was validated according to parameters in the RE 899/2003. As result, it was obtained from the AcOEt fraction the compound Kb1 (2.7 mg). Until this moment, the basic nucleus was characterized by UV analysis using shift reagents. The partial chemical structure of the compound Kb1 was identified as a flavonol, containing hydroxyls in 3 , 4 position (ring A), 5 and 7 free (ring B) and a replacement of the C3 hydroxyl by a sugar. As the analysis were performed in the HPLC coupled to a DAD, we observed that the UV spectrum of the major peaks of EH from K. brasiliensis shown similar UV spectrum. According to the literature, it has been reported the presence of patuletin glycosydes derivatives in the leaves of this species. Therefore, it is suggested that the compound Kb1 is glycosylated patuletin derivative. Probably the sugar (s) unit(s) are linked in the C3 in the C ring. . Regarding the development of HPLC analytical method, the system used consists of phase A: water: formic acid (99,7:0,3, v / v) and phase B: methanol: formic acid (99,7:0,3, v / v), elution gradient of 40% B - 58% B in 50 minutes, ccolumn (Hichrom ?) C18 (250x4, 0 mm, 5 ?m), flow rate 0.8 mL / min, UV detection at 370 nm, temperature 25 ? C. In the analysis performed with the co-injection of thecompound Kb1 + HE of K. brasiliensis was observed that it is one of the major compounds with a retention time of 12.47 minutes and had a content of 15.3% in EH of leaves from K. brasiliensis. The method proved to be linear, precise, accurate and reproducible. According to these results, it was observed that compound Kb1 can be used as a chemical marker of EH from leaves of K. brasiliensis, to assist in quality control of drug plant and its derivatives / Kalanchoe brasiliensis Cambess (Crassulaceae), conhecida popularmente como sai?o, coirama branca e folha grossa, ? origin?ria do Brasil e comumente encontrada de S?o Paulo at? a Bahia, principalmente na zona litor?nea. Quanto ? atividade microbiol?gica, estudos pr?-cl?nicos foram realizados no sentido de investigar a atividade antiinflamat?ria, para os extratos obtidos a partir folhas e/ou partes ?reas de K. brasiliensis. Em rela??o ? constitui??o qu?mica, h? relatos principalmente da presen?a de flavonoides nas folhas da esp?cie. Embora seja uma esp?cie utilizada amplamente na medicina tradicional no Brasil, n?o h? monografia que especifique os par?metros de qualidade da droga vegetal. Dentro deste contexto, o presente trabalho tem como objetivo caracterizar e a quantificar os marcadores qu?micos do extrato hidroetan?lico (EH) das folhas de K. brasiliensis, que possam ser utilizados no controle de qualidade de derivados obtidos a partir dessa esp?cie. A metodologia do estudo foi dividida em duas partes: i. Estudo fitoqu?mico: fracionamento, isolamento e caracteriza??o de marcador(es) qu?mico(s) do extrato hidroetan?lico das folhas de K. brasiliensis; ii.Desenvolvimento e valida??o de metodologia anal?tica por Cromatografia L?quida de Alta Efici?ncia (CLAE) para quantificar (o)s marcador(es) qu?mico(s) do EH. i. O EH 50% foi obtido pelo m?todo de turboextra??o. Em seguida foi submetido a uma parti??o l?quido-l?quido, obtendo-se as fra??es diclorometano, n-butanol e acetato de etila (FAcOEt). A fra??o acetato de etila foi selecionada para dar continuidade ao fracionamento, e por apresentar um perfil qu?mico rico em flavonoides, uma vez que esses metab?litos parecem estar relacionados ? atividade farmacol?gica. ii. Para a an?lise quantitativa, o EH foi analisado por CLAE, utilizando diferentes m?todos. Ap?s selecionado o m?todo mais adequado, que apresentou picos sim?tricos e resolu??o satisfat?ria, foi realizada a valida??o segundo par?metros da RE No 899/2003. Como resultados, foi obtido a partir da fra??o acetato de etila o composto Kb1 (2,7mg). Neste composto, foi caracterizado o n?cleo b?sico flavono?dico por an?lise por UV utilizando reagentes de deslocamento. O composto Kb1 foi identificado como um flavonol, com hidroxilas em 3 , 4 (anel B) e 5 e 7 livres (anel A), al?m de uma hidroxila substitu?da em C3. Como foi utilizado a CLAE acoplada a um detector de arranjo de diodos, foi poss?vel observar que os espectros UV dos picos majorit?rios do EH de K. brasiliensis s?o similares. Em estudos anteriores foi descrita a presen?a de derivados glicosilados da patuletina nas folhas dessa esp?cie. Portanto, sugere-se que Kb1 seja um derivado glicosilado na posi??o C-3 da patuletina. Quanto ao desenvolvimento anal?tico por CLAE, o sistema utilizado foi constitu?do de fase A: ?gua:?cido f?rmico (99,7:0,3, v/v) e fase B: metanol:?cido f?rmico (99,7:0,3, v/v) em elui??o gradiente de 40%B - 58%B em 50 minutos, coluna cromatogr?fica (Hichrom) C18 (250x4,0mm, 5?m), fluxo 0,8 mL/min, detec??o em UV a 370 nm, temperatura de 25oC. Por meio de co-inje??o do padr?o + extrato foi observado que Kb1 trata-se de um dos compostos majorit?rios, com tempo de reten??o de 12,47 minutos e apresentou um teor de 15,3% no EH de K. brasiliensis O m?todo apresentou-se linear, preciso, exato e reprodut?vel. De acordo com os resultados obtidos, foi observado que Kb1 pode ser utilizado como marcador qu?mico do EH das folhas de K. brasiliensis, para auxiliar no controle de qualidade da droga vegetal e seus derivados

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-04-03T19:41:30Z No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-04-05T19:31:53Z (GMT) No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) / Made available in DSpace on 2017-04-05T19:31:53Z (GMT). No. of bitstreams: 1 SarahPollyanaDiasDosSantos_DISSERT.pdf: 10111393 bytes, checksum: 44cc6b08e8b21238733661e330e1b811 (MD5) Previous issue date: 2016-07-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Aspidosperma pyrifolium Mart., esp?cie nativa da caatinga com grande ocorr?ncia no nordeste, conhecida popularmente como pereiro, utilizada na medicina popular no tratamento de doen?as e enfermidades. Consta na literatura diversos trabalhos realizados, relatando a presen?a de classes de compostos org?nicos como alcaloides, entre outras, que seriam respons?veis por propriedades terap?uticas existentes nessa esp?cie. O estudo fitoqu?mico de A. pyrifolium, das fra??es da semente e cascas do fruto, utilizando m?todos cromatogr?ficos cl?ssicos e purifica??o por CLAE, nos permitiu o isolamento de tr?s compostos. A partir da fra??o hex?nica das cascas do fruto (APCF-H) foi isolado o triterpeno ?cido betul?nico, n?o relatado anteriormente na esp?cie A. pyrifolium. Das fra??es diclorometano (APSE/DCM) e acetato de etila 50% dicloromentano (APSEA/50% DCM) das sementes foi poss?vel o isolamento do alcaloide plumerano pentac?clico aspidospermina, do alcaloide ?-carbolina glicosilado Cordifolina, n?o descrito anteriormente na esp?cie e outro composto do tipo ?-carbolina que n?o possui uma estrutura elucidada at? o momento. O extrato das sementes fra??o diclorometano (APSE/DCM) foi analisado por CG/EM (cromatografia gasosa acoplada a espectrometria de massa) e seus dados foram comparados com dados na literatura em banco de dados (NIST08.LIB), foi identificada a presen?a dos alcaloides 1-acetil-aspidospermidina, aspidospermina e pirifolina, dentre outros compostos minorit?rios. A avalia??o da atividade antioxidante, pelo m?todo sequestrador de radical livre de DPPH (2,2-difenil-1picril-hidrazila), da fra??o APSE/DCM apresentou um percentual de atividade antioxidande m?ximo (AA% m?ximo) de 83,68%, com concentra??o inibit?ria 50% (IC50) 133,4867 ? 2,22. A fra??o das sementes fra??o diclorometano (APSE/DCM) foi avaliada em rela??o ? toxicidade contra Artemia salina, tendo em vista que j? existiam relatos da toxicidade nas folhas de A. pyrifolium, comprovamos a toxicidade aguda nas fra??o das sementes diclorometano com concentra??o letal m?dia (CL50) em 24 horas de 1,1524 ?g.ml-1, sendo consideradas t?xicas as esp?cies que apresentam CL50 inferior a 1000 ?g.ml-1. Na segunda parte do trabalho foi realizado um levantamento bibliogr?fico de alcaloides do tipo ?-carbolina de ocorr?ncia natural, com suas estruturas, atividades relatadas e um banco de dados com seus deslocamentos qu?micos em RMN 1H e 13C, totalizando 169 alcaloides descritos na literatura, do per?odo de 1992 a 2016, agrupados em quatro principais grupos, denominados ?-carbolina, dihidro-?-carbolina, tetrahidro-?-carbolina e os ?-carbolinas divergentes, este levantamento auxiliar? na elucida??o estrutural de novas mol?culas. / Aspidosperma pyrifolium Mart., a native species of the caatinga with great occurrence in the northeast, known perpularmente as pereiro, used in popular medicine in the treatment of diseases and diseases. There are several studies carried out in the literature, reporting the presence of classes of organic compounds such as alkaloids, among others, that would be responsible for the therapeutic properties of this species. The phytochemical study of A. pyrifolium, seed fractions and fruit peels using classical chromatographic methods and purification by HPLC allowed us to isolate three compounds. From the hexanic fraction of fruit peels (APCF-H) was isolated the betulinic acid triterpene, not previously reported in the A. pyrifolium species. From the dichloromethane (APSE/DCM) and ethyl acetate 50% dichloromethane (APSEA / 50% DCM) fractions of the seeds, it was possible to isolate the pentacyclic pentameric aspidospermine alkaloid from the glycosylated ?-carboline alkaloid Cordifolina, not previously described in the species and another compound of the ?-carboline type that does not have a structure elucidated until the moment. The extract of the seeds dichloromethane fraction (APSE/DCM) was analyzed by GC/MS (gas chromatography coupled to mass spectrometry) and its data were compared with data in the literature in database (NIST08.LIB), the presence of the alkaloids 1-acetyl-aspidospermidine, aspidospermine and pyrifoline, among other minor compounds. The antioxidant activity evaluation of the APSE/DCM fraction showed a percentage of maximum antioxidant activity (AA% maximum) of 83.68%, with the free radical sequestration method of DPPH (2,2-diphenyl-1-picryl-hydrazila). Inhibitory concentration 50% (IC50) 133.4867 ? 2.22. The fraction of the seeds dichloromethane fraction (APSE/DCM) was evaluated in relation to the toxicity against Artemia salina, considering that there were already reports of the toxicity in the leaves of A. pyrifolium, we verified the acute toxicity in the seed dichloromethane fraction with average lethal concentration (CL50) in 24 hours of 1.1524 ?g.ml-1, species with CL50 less than 1000 ?g.ml-1 being considered toxic. In the second part of the work a bibliographic survey of naturally occurring ?-carboline alkaloids with their structures, reported activities and a database with their chemical displacements in 1H and 13C NMR were carried out, totaling 169 alkaloids described in the literature. From 1992 to 2016, grouped into four main groups, named ?-carboline, dihydro-?-carboline, tetrahydro-?-carboline and the divergent ?-carbolines, this survey will aid in the structural elucidation of new molecules.

The bacteria Streptococcus pyogenes produces a multitude of proteins which interact with and alter the functions of the host immune system. Two such proteins, Endoglycosidase S (EndoS) and Immunoglobulin G-degrading enzyme from S. pyogenes (IdeS) are able to specifically alter the effector functions of immunoglobulin G (IgG). EndoS is a glycoside hydrolase which removes the conserved <i>N</i>-linked glycan from IgG Fc whereas IdeS is a cysteine protease that cleaves the exible protein hinge of IgG. The activity of both proteins results in the reduced ability of IgG to elicit immune responses through Fc receptor binding and complement activation. Amongst other applications, both EndoS and IdeS are actively being explored as new therapeutics for IgG-mediated autoimmune diseases. Given the therapeutic potential of EndoS and IdeS, experiments were designed to investigate the structural and functional characteristics of these enzymes in an effort to understand their specficity for and activity against IgG. Here, bioinformatic and biophysical characterisation of EndoS identified subdomains outside of the catalytic domain which contribute to glycoside hydrolase activity. The substrate specificity of EndoS was also explored and showed that EndoS hydrolyses a broad range of glycans from the IgG scaffold. EndoS was also shown to have activity against alternative glycoprotein substrates, however, this non-specific activity was negligible in the context of whole serum. The effect of EndoS-mediated deglycosylation on the structure of the IgG Fc domain was explored using both X-ray crystallography and small-angle X-ray scattering. Small angle X-ray scattering was also used to characterise both EndoS and IdeS in complex with IgG Fc. Solution-state models of each complex were produced providing preliminary data towards how these enzymes interact with IgG. Overall, the results presented here contribute to our understanding of these enzymes which is of importance as they go forward into clinical applications.

Bacteria of the genus Streptococcus are common asymptomatic colonisers of humans and animals. As opportunistic pathogens they can however, depending on their host’s immune status and other circumstances, cause mild to very severe infections. Streptococci are highly intertwined with specific host species, but can also cause zoonosis or anthroponosis in more uncommon hosts. Prolonged and reoccurring infections require immune evasion strategies to circumvent detection and eradication by the host’s immune defence. A substantial part of the immune defence against bacterial pathogens is mediated by immunoglobulins. This thesis is based on work to identify and characterise immunoglobulin degrading enzymes secreted by different Streptococcus species as a means to sabotage and evade antibody-mediated immune responses. Stoichiometric and kinetic analysis of the IgG degrading enzyme IdeS from the important human pathogen S. pyogenes revealed that IdeS cleaves IgG, opposed to previous publications, as a monomer following classical Michaelis-Menten kinetics. The IdeS homologue of S. suis, IdeSsuis, did however not cleave IgG, but was highly specific fo rporcine IgM. S. suis was found to possess yet another protease, IgdE, capable of cleaving porcine IgG. Both of these proteases were shown to promote increased bacterial survival in porcine blood during certain conditions. IgdE is the founding member of a novel cysteine protease family (C113). Novel streptococcal members of this protease family were shown to specifically degrade certain IgG subtypes of the respective Streptococcus species’ main host. The observed substrate specificity of IgdE family proteases reflects the host tropism of these Streptococcus species, thereby giving insights into host-pathogen co-evolution. The abundance of immunoglobulin degrading enzymes among Streptococcus species indicates the importance of evasion from the antibody mediated immune responses for streptococci. These novel identified immunoglobulin degrading enzymes of the IdeS and IgdE protease families are potential valid vaccine targets and could also be of biotechnological use.

Made available in DSpace on 2016-04-28T14:57:35Z (GMT). No. of bitstreams: 1 2008 - Rhadyson Reinaldo Silva do Nascimento.pdf: 1733739 bytes, checksum: b1070eca267f85de92499d2e245add23 (MD5) Previous issue date: 2008-09-03 / Cowpea (Vigna unguiculata L. Walp.) is a very important crop in Brazil, mainly in the north and northeast regions of the country, and is gaining importance in the southeast region. In this crop, nematodes are a great threat, because they can cause yield decrease. Information about these pathogens is very few. In a first approach this work aimed at surveying for the occurrence of plant parasitic nematodes that are associated to cowpea in the State of Rio de Janeiro, through sampling achieved in some locations of Nova Igua?u, Mag?, Serop?dica, S?o Francisco de Itabapoana, and Itagua?, where cowpea is grown cultivated. Eight genera of plant parasitic nematodes we found, besides others, that are free leving that were not identified. At the level of species M. incognita and Rotylenchulus reniformis were identified. Nematode identification was done on the bases of morphometric characteristics descriptive keys M. incognita by juvenile (J2 ) and males characteristics and perineal female patterns and esterases profile. In this step, surveys were carried out aiming at the detection of esterases activity for a population of R. reniformis. Protocols routinely used in isoezymes analysis were tested and showed negative results. On the other hand, assays using PCR tubes with the objective to identify esterase activity demonstrated that it was positive for R. reniformis. Modifications in the common protocols, such as changes in buffer solution at the and steps of the procedures, as well as adjusting pH near to neutrality enabled the establishment of a three band pattern of esterase in the studied populations of R. reniformis. In another phase of this work, six cowpea lines in advanced phase of improvement from determining their behavior when inoculated with M. incognita race 1, M. javanica and R. reniformis. The tomato cv. TRural was used as susceptible pattern. The experiments were carried out under greenhouse condition, using 5,000 eggs + juveniles per experimental plots, as inoculum level; for Meloidogyne species evaluation was done 50 days after nematodes inoculations based in the following parameters: galls indices (IG), egg mass indices (IMO) and reproductions rate (FR), calculated by the relation Pf/Pi ( Pf = final populations and Pi= start population). Among the tested lines, five of then RJ 04-04, RJ 04-08, RJ 04-26, RJ 04-48 and RJ 04-65 showed to be highly resistant and line RJ 04-29 was moderately resistant to M. incognita race 1. In the case of M. javanica all lines were highly susceptible. For evaluation of R. reniformis it was used as inoculum and "pure" population multiplied vegetation home. It was inoculated 5000 eggs and juvenile of males and of immature females. As parameter for evaluation of the resistance of the cowpea lineages to the nematode, the reproduction factor was used (Fr) appraised to the 45 days after inoculation. It was verified that all of the tested lineages were considered susceptible; to cultivate "Costel?o" included as witness showed equal behavior. / O caupi (Vigna unguiculata Walp.) ? uma cultura de grande import?ncia para o pa?s, principalmente na regi?o Norte e Nordeste e vem ganhando espa?o na regi?o Sudeste. Nesta cultura, os nemat?ides possuem influ?ncia significativa, chegando ao ponto de limitar a sua produ??o. As informa??es sobre estes pat?genos no Estado do Rio de Janeiro s?o escassas. Assim, numa primeira etapa, este trabalho teve por objetivo fazer um levantamento de nemat?ides fitoparasitas associados a cultura do caupi no Estado do Rio de Janeiro, por meio de coletas realizadas em alguns munic?pios produtores de caupi cultura. Foram encontrados, um total de oito g?neros de nemat?ides fitoparasitas, al?m de outros de vida livre n?o identificados. Em n?vel de esp?cie foram identificadas Meloidogyne incognita ra?a 2 e Rotylenchulus reniformis. As identifica??es foram efetuadas a partir de caracter?sticas morfom?tricas e aux?lio de chaves descritivas. M. incognita foi identificada pelas caracter?sticas dos juvenis (J2) e de machos, configura??o do modelo perineal das f?meas e perfil de esterases. Foram realizados ensaios com o objetivo de detec??o de atividade de esterase, para uma popula??o do nemat?ide R. reniformis. Para tanto foram testados protocolos rotineiramente usados em an?lises isoenzim?ticas, que apresentaram resultados negativos. Entretanto, ensaios montados em microtubos para PCR, evidenciaram atividade ester?sica para R. reniformis. Finalmente, modifica??es nos protocolos usuais como mudan?as na concentra??o da solu??o tamp?o de revela??o, assim como a fixa??o do pH da mesma pr?ximo a neutralidade, permitiram estabelecer um padr?o com tr?s bandas distintas de esterases na popula??o de R. reniformis estudada. Em uma segunda etapa deste trabalho, seis linhagens de caupi em fase avan?ada de melhoramento para resist?ncia a viroses, foram testadas visando determinar a rea??o das mesmas quando inoculadas com M. incognita ra?a 1, ou M. javanica, ou R. reniformis. A cultivar de tomate TRural foi usada como testemunha suscet?vel nos testes com as duas esp?cies de nemat?ide das galhas radiculares. Os experimentos foram realizados em casa de vegeta??o e usou-se uma carga de in?culo de 5000 ovos + juvenis/repeti??o; para as esp?cies de Meloidogyne a avalia??o ocorreu 50 dias ap?s inocula??o e os par?metros usados foram: ?ndice de galhas (IG), ?ndice de massa de ovos (IMO) e fator de reprodu??o (Fr) calculado pela rela??o Pf/Pi (Pf = popula??o final e Pi = popula??o inicial). Das cinco linhagens testadas, RJ 04-04, RJ 04-08, RJ 04-26, RJ 04-48 e RJ 04-65 comportaram-se como altamente resistentes e a linhagem RJ 04-29 como moderadamente resistente para M. incognita ra?a1. Enquanto que, para M. javanica, todas as linhagem foram altamente suscet?veis. Para avalia??o de R. reniformis foi usado como in?culo uma popula??o pura multiplicada em casa de vegeta??o. Inoculou-se 5000 ovos e/ ou juvenis de machos e de f?meas imaturas. Como par?metro para avalia??o da resist?ncia das linhagens de caupi ao nemat?ide, foi usado o fator de reprodu??o (Fr) avaliado aos 45 dias ap?s inocula??o. Verificou-se que todas as linhagens testadas foram consideradas suscet?veis; a cultivar Costel?o inclu?da como testemunha mostrou igual comportamento.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-08-26T14:00:38Z No. of bitstreams: 1 2011- C?ntia Let?cia da Silva Rosa.pdf: 3366742 bytes, checksum: 280ece7f9ee6afde0d927d4a7dd0c3ef (MD5) / Made available in DSpace on 2016-08-26T14:00:38Z (GMT). No. of bitstreams: 1 2011- C?ntia Let?cia da Silva Rosa.pdf: 3366742 bytes, checksum: 280ece7f9ee6afde0d927d4a7dd0c3ef (MD5) Previous issue date: 2011-03-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This study aimed to evaluate accessions of tomato 'heirloom' whose cultivation is still little known in Brazil, grown organically, and its physical-chemical, nutritional and sensory characteristics after processing to obtain concentrated pulp. We used the non-hybrid tomatoes (Lycopersicum esculentum Mill) San Marzano, Chico Grande, Amish Paste and called EUA 05. The fruits were produced in the period from May to September 2010, the sector of Horticulture Department of Plant Science, of UFRRJ. In fresh fruit were analyzed total soluble solids (TSS), acidity (TTA), pH, quantification and identification of carotenoids, color of skin and pulp, texture instrumental and analysis of pesticides. Access San Marzano was not processed because of bacterial contamination in the field. The process was composed of the stages of pre-washing, washing / screening, milling, pulping, concentration, filling, exhaustion, closing, heat treatment / cooling and storage. It were added in the formulation of 1% salt, 0.4% sugar and citric acid for pH correction for microbial control. It was performed viscosity and pre-commercial sterility analyses in addition to those cited in the fresh tomato, both in the pulps in this study as six commercial brands on the market. For the sensorial analysis of the product manufactured was performed the Quantitative Descriptive Analysis (QDA) with trained judges and acceptance test with consumers. With respect to the TSS in tomatoes fresh, the Chico Grand and San Marzano showed the highest values (5.2? Brix). Access Chico Grande showed greater balance between acidity and sugar content in fruit, established by the TSS / TA (? Brix /%) of 19.2. Regarding the content of lycopene, access San Marzano had a higher average level of 6029 ?g/100g and more red coloration observed in the amount of a *= 29.68. No pesticide residues were found in the samples for organochlorine and organophosphorus pesticides. Results for the SST of pulps showed that accessions Amish Paste and EUA 05 stood out with values of 10.1 ? Brix. Regarding ATT access Amish Paste showed higher citric acid (1.445 g/100g) and lower pH value (3.66), which can lead to an impairment of sensory quality. The efficiency of the processing of pulp was 73% for access Chico Grande, Amish Paste 72% and 81% for EUA 05. After analyzing the lycopene content of the samples, we observed that the sample Amish Paste (3346.0 ?g/100g) and EUA 05 (3375.0 ?g/100g) showed the highest values with no statistical difference between them, corroborating the red color represented by lower values of hue angle. With respect to viscosity, the samples Chico Grande (246cP) and EUA 05 (235cP) stood out by the highest maximum viscosity. The results found in ADQ characterized the pulps by the attributes of astringency, taste salty, sour and acid flavor, and the commercialpulps by the attributes sweet aroma and sweet taste, flavor and aroma of tomato pulp. With respect to acceptance, the lowest scores were attributed to pulp in this study. However, samples Chico Grande (4.41) and the EUA 05 (4.48) had the highest score for overall acceptability with statistically significant differences with regard to the sample Amish Paste (3.06). It can be concluded with respect to fresh tomatoes that tomato 'heirloom' evaluated showed good quality in relation to balance acidity/sugar, and lycopene content. In pulps prepared from the three accessions 'heirloom' studied, concentrated pulp obtained from tomato Amish Paste and USA 05 showed the best characteristics for the food industry, with respect to soluble solids, lycopene content and red color. However, access Chico Grande showed higher viscosity when compared to other accessions. After the sensory analysis concluded that there was a rejection by the majority of consumers with regard to all the pulps obtained from tomatoes 'heirloom' due to significant changes in aroma and taste sour, salty taste as well as during the preparation / Este trabalho teve como objetivo geral avaliar acessos de tomate ?heirloom? cujo cultivo ainda ? pouco conhecido no Brasil, cultivados no sistema org?nico, e suas caracter?sticas f?sicoqu?micas, nutricionais e sensoriais ap?s o processamento para obten??o de polpa concentrada. Foram utilizados os tomates n?o h?bridos (Lycopersicum esculentum Mill) San Marzano, Chico Grande, Amish Paste e o denominado EUA 05. Os frutos foram produzidos no per?odo de maio a setembro de 2010, no setor de Horticultura do Departamento de Fitotecnia, da UFRRJ. Os frutos in natura foram caracterizados pelas an?lises de s?lidos sol?veis totais (SST), acidez total titul?vel (ATT), pH, quantifica??o e identifica??o dos caroten?ides, cor da casca e da polpa, textura instrumental e an?lise de pesticidas. O processamento e as an?lises foram realizados na Embrapa Agroind?stria de Alimentos, Rio de Janeiro-RJ. O processamento foi composto pelas etapas de pr?-lavagem, lavagem/sele??o, tritura??o, despolpamento, concentra??o, envase, exaust?o, fechamento, tratamento t?rmico/resfriamento e armazenamento. O acesso San Marzano n?o foi utilizado devido ? contamina??o bacteriana no campo. Foram adicionados na formula??o 1% de sal, 0,4% de a??car e ?cido c?trico para corre??o do pH para controle microbiol?gico. As mesmas an?lises realizadas no tomate in natura e ainda as an?lises de viscosidade, coliformes a 45?C, salmonelas, pr?-esterilidade comercial e avalia??o sensorial foram realizadas nas polpas concentradas e em seis marcas comerciais do mercado. Para realiza??o da an?lise sensorial utilizou-se a An?lise Descritiva Quantitativa (ADQ) com equipe treinada e Teste de Aceita??o com consumidores. Com rela??o ao teor de SST nos tomates in natura os acessos Chico Grande e San Marzano apresentaram os maiores valores (5,2 ?Brix). O acesso Chico Grande apresentou maior balan?o entre acidez e o teor de a??cares no fruto, estabelecido pela rela??o SST/ATT (?Brix/%) de 19,2. Com rela??o ao teor de licopeno o acesso San Marzano apresentou maior teor m?dio de 6029 ?g/100g, bem como colora??o mais vermelha observada pelo par?metro instrumental a*=29,68. N?o foram encontrados res?duos de agrot?xicos organofosforados e organoclorados em nenhuma das amostras. Nas polpas concentradas os resultados encontrados para a an?lise SST demonstraram que os acessos Amish Paste e EUA 05 se destacaram com valores de 10,1?Brix. O acesso Amish Paste apresentou maior valor de acidez (1,445g/100g), bem como menor valor de pH (3,66), o que levou a um comprometimento da sua aceita??o sensorial. O rendimento ap?s despolpamento dos tomates foi de 73% para o acesso Chico Grande, 72% para Amish Paste e de 81% para EUA 05. Ap?s a an?lise do teor de licopeno das amostras, observou-se que a amostra Amish Paste (3346,0?g/100g) e EUA 05 (3375,0?g/100g) apresentaram os maiores valores sem diferen?a estat?stica entre si, corroborando com a colora??o vermelha representada pelos menores valores do ?ngulo hue (?h). As polpas Chico Grande (246cP) e EUA 05 (235cP) se destacaram pelos maiores valores de viscosidade m?xima. A avalia??o sensorial (ADQ) caracterizou as polpas obtidas pelos atributos adstring?ncia, gosto salgado, gosto ?cido e aroma ?cido; e as polpas comerciais pelos atributos aroma doce, gosto doce, sabor e aroma caracter?stico de polpa de tomate. Com rela??o ? aceita??o, as menores notas foram atribu?das ?s polpas deste estudo. No entanto, as amostras Chico Grande (4,41) e EUA 05 (4,48) apresentaram a maior nota de aceita??o global com diferen?a estat?stica significativa com rela??o ? amostra Amish Paste (3,06). Pode-se concluir com rela??o ao tomate in natura que os acessos de tomate ?heirloom? avaliados apresentaram boa qualidade com rela??o ao equil?brio acidez/a??cares e teores de licopeno. Nas polpas elaboradas dos tr?s acessos ?heirloom? estudados, a polpa concentrada obtida do tomate EUA 05 e Amish Paste apresentaram melhores caracter?sticas para a ind?stria de alimentos, com rela??o ao teor de s?lidos sol?veis, teor de licopeno e colora??o vermelha. No entanto, o acesso Chico Grande apresentou valores maiores de viscosidade quando comparados aos outros acessos. Ap?s an?lise sensorial concluise que houve uma rejei??o pela maioria dos provadores com rela??o a todas as polpas obtidas dos tomates ?heirloom? devido a altera??es significativas no aroma e sabor ?cido, bem como no gosto salgado, decorrentes de sua formula??o.

Made available in DSpace on 2014-12-17T14:03:41Z (GMT). No. of bitstreams: 1 VanessaOS.pdf: 1126102 bytes, checksum: dba464c890585c6be32a3fd2dc600a81 (MD5) Previous issue date: 2006-06-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Sulfated Polysaccharides with unique chemical structures and important biological activities has been found in a diversity of sea invertebrates. For that, to exist a huger interest on the biotechnology field in the research theses sulfated compounds isolated from sea organisms. Despite the privileged brazilian position for these compounds attainment, there are still a few scientific informations about the isolated substances and their biological activities. A head the displayed, the present work has for objectives, to evaluate the pharmacological properties of the glycosaminoglycans isolated from the sea shrimp Litopenaeus schimitti on homeostasis, blood coagulation, leukocytes migration and platelet/leukocyte adhesion. For this, yhe glycosaminoglycans were extracted from crustacean tissues by proteolysis, fractionation with acetone and later submitted to pharmacological assays. The crustacean tissues showed compounds heparin-like, with anticoagulant activity of 45 IU/mg and 90 IU/mg, respectively. These molecules showed low residual hemorrhagic effects in the tested concentration (100 ?g/mL), when compared to unfractionated commercial heparin (UFH). Another dermatan sulfate-like compound, predominately constituted for disulfated disaccharides, was isolated from crustacean abdomen. This compound showed an efficient effect on leukocytes migration inhibition, in the concentration of 15 ?g/mL, reducing the cellular infiltration in 65% when compared to the controlled animals. In this same concentration, the DS reduced in 60% the protein concentration of the peritoneal exudates. In the concentration, this compound of 0.5 mg/mL, it was capable to reduce in 40% platelet/leukocytes adhesion. Our data demonstrate that these sulfated polysaccharides isolated from the shrimp L. schimitti will can be used as bioactive compounds, appearing as active principles for pharmacological development, anticoagulants and inflammatory response regulators / Polissacar?deos sulfatados com caracter?sticas estruturais distintas e importantes atividades biol?gicas t?m sido encontrados em uma diversidade de invertebrados marinhos. Por isso existe um grande interesse no campo da biotecnologia na pesquisa destes compostos sulfatados isolados de organismos aqu?ticos. No entanto, apesar da posi??o privilegiada do Brasil para a obten??o destes compostos, ainda s?o poucas as informa??es cient?ficas sobre as subst?ncias isoladas e suas atividades biol?gicas. Diante do exposto, este presente trabalho teve por objetivos avaliar os potenciais farmacol?gicos dos glicosaminoglicanos (GAGs) isolados do camar?o marinho Litopenaeus schimitti, sobre a hemostasia, coagula??o sang??nea, migra??o leucocit?ria e ades?o celular. Para isso os GAGs foram extra?dos dos tecidos do crust?ceo mediante prote?lise, fracionamento com acetona e posteriormente submetidos aos ensaios farmacol?gicos. Os tecidos do crust?ceo, abd?men e cefalot?rax, apresentaram compostos semelhantes ? heparina (heparin?ides) com atividade anticoagulante de 45 UI/mg e 90 UI/mg, respectivamente. Estas mol?culas apresentaram baixo efeito hemorr?gico residual na concentra??o de 100 ?g/mL, quando comparada com a heparina comercial n?o fracionada (HNF). Um outro composto semelhante ao dermatam sulfato (DS), constitu?do predominantemente por dissacar?deos dissulfatados foi isolado do abd?men do crust?ceo. Este composto apresentou, na concentra??o de 15 ?g/?L, uma inibi??o significativa (P<0.01) da migra??o leucocit?ria, reduzindo a infiltra??o celular em 65% quando comparado com os animais controle. Nessa mesma concentra??o o DS reduziu em 60% a concentra??o de prote?nas do lavado peritonial. As an?lises qualitativas da composi??o celular do exudato peritonial foram similares ao encontrado para os animais controles em todas as concentra??es testadas. Na concentra??o de 0,5 mg/mL foi capaz de reduzir em 40% a ades?o das plaquetas aos leuc?citos. Os dados obtidos demonstram que estes polissacar?deos sulfatados isolados do camar?o L.schimitti podem vir a ser utilizados como compostos bioativos, podendo surgir como princ?pios ativos para o desenvolvimento de f?rmacos, anticoagulantes e moduladores da resposta inflamat?ria

[ES] Actualmente, existe una gran variedad de tecnologías de distribución de contenidos multimedia, de dispositivos de consumo, así como de tipos de contenidos. En cuanto a tecnologías de distribución, dichos contenidos se pueden enviar bien a través de redes broadcast, como las redes DVB, o bien a través de redes broadband, como las redes IP (por ejemplo, a través de Internet). La coordinación y convergencia entre los dos tipos de tecnologías para distribuir contenidos relacionados ofrece un gran potencial a los proveedores de contenidos, así como grandes beneficios a los consumidores (una prueba de ello es el estándar Hybrid Broadcast Broadband TV o HbbTV). Sin embargo, la recepción de contenidos relacionados procedentes de diferentes redes híbridas (broadcast/broadband) conlleva una variabilidad de retardos significativa, por lo que se deben superar una serie de retos importantes, como, por ejemplo, la necesidad de mecanismos que garanticen un consumo sincronizado de dichos contenidos, bien en un mismo dispositivo o bien en distintos dispositivos (locales o remotos). Se han especificado, implementado y evaluado soluciones de sincronización de los procesos de reproducción de contenidos híbridos (prevenientes de redes broadcast y/o broadband) tanto en entornos multidispositivo, entre dispositivos locales (Inter-Device Synchronization o IDES), como en entornos multiusuario, entre destinatarios distribuidos en red (Inter-Destination Media Synchronization o IDMS), incluyendo la sincronización de efectos multisensoriales. Se ha diseñado e implementado una plataforma extremo-a-extremo para la generación, distribución (adaptativa) y consumo de contenidos híbridos relacionados, tanto en un mismo dispositivo como en múltiples dispositivos locales (por ejemplo, escenario multipantalla) o remotos (por ejemplo, TV Social). Se han generado y preparado contenidos multimedia tradicionales (2D) y omnidireccionales (360), enriquecidos con efectos multisensoriales que mejoran el grado de realismo y la inmersividad de los usuarios. La plataforma es compatible con el estándar HbbTV y, además, incluye soluciones eficientes para aspectos clave que no están especificados en dicho estándar, pero que son necesarios para la implementación satisfactoria de servicios de TV híbridos y multidispositivo en la misma, como, por ejemplo: la necesidad de un buffer para el contenido broadcast (existente y opcional hasta la especificación HbbTV 2.0.2); mecanismos de señalización, asociación y descripción de contenido disponible y relacionado con el contenido principal; mecanismos de coordinación e interacción entre los dispositivos de consumo; o soluciones de sincronización para alinear en el tiempo el consumo de contenidos de forma satisfactoria. Los resultados obtenidos en la evaluación realizada muestran que el nivel de sincronización adquirido entre los procesos de reproducción de los diversos tipos de contenido disponibles a través de distintas tecnologías de transmisión alcanza unos valores suficientes para proporcionar al usuario un consumo con un nivel de asincronías imperceptible. Dicha precisión alcanzada ha resultado en una QoE para el consumidor muy satisfactoria, ofreciéndole al mismo un completo ecosistema de consumo de contenidos que incluye tanto contenido multimedia tradicional, como contenido omnidireccional y mulsemedia. / [CA] Actualment, hi ha una gran varietat de tecnologies de distribució de contingut multimèdia, dispositius de consum, així com tipus de contingut. Pel que fa a les tecnologies de distribució, aquest contingut es pot lliurar a través de xarxes de difusió, com ara xarxes DVB, o a través de xarxes de banda ampla, com les xarxes IP (per exemple, a través d'Internet). La coordinació i convergència entre els dos tipus de tecnologies per distribuir continguts relacionats ofereix un gran potencial als proveïdors de continguts, així com grans beneficis per als consumidors (una prova d'això és l'estàndard Hybrid Broadcast Broadband TV o HbbTV). Tanmateix, la recepció de continguts relacionats des de diferents xarxes híbrides (emissió/banda ampla) comporta una important variabilitat de retard, per la qual cosa s'han de superar una sèrie de reptes importants, com la necessitat de mecanismes que garanteixin el consum sincronitzat d'aquests continguts, ja siga en el mateix dispositiu. o en diferents dispositius (locals o remots). S'han especificat, implementat i evaluat solucions de sincronització per a processos de reproducció de continguts híbrids (xarxes de broadcast i/o de banda ampla) tant en entorns multidispositiu, entre dispositius locals (Inter-Device Synchronization o IDES), com en entorns IDMS (Inter-Destination Media Synchronization o IDMS), incloent la sincronització d'efectes multisensorials. S'ha dissenyat i implementat una plataforma d'extrem a extrem per a la generació, distribució (adaptativa) i consum de contingut híbrid relacionat, tant en un sol dispositiu com en múltiples locals (p. ex., escenari multipantalla) o remots (p. ex., Social TV). dispositius. S'han generat i preparat contingut multimèdia tradicional (2D) i omnidireccional (360), enriquit amb efectes multisensorials que milloren el grau de realisme i immersivitat dels usuaris. La plataforma és compatible amb l'estàndard HbbTV i, a més, inclou solucions eficients per a aspectes clau que no s'especifiquen en aquest estàndard, però que són necessaris per a la implantació amb èxit de serveis de televisió híbrids i multidispositiu en el mateix, com ara: la necessitat per a un buffer per a continguts de difusió (existent i opcional fins a HbbTV 2. 0.2); mecanismes de senyalització, associació i descripció dels continguts disponibles i relacionats amb el contingut principal; mecanismes de coordinació i interacció entre dispositius de consum; o solucions de sincronització per alinear en el temps el consum de continguts de manera satisfactòria. Els resultats obtinguts en l'avaluació realitzada mostren que el nivell de sincronització adquirit entre els processos de reproducció dels diferents tipus de continguts disponibles a través de diferents tecnologies de transmissió assoleix valors suficients per proporcionar a l'usuari un consum amb un nivell d'asincronies imperceptibles. La precisió aconseguida ha donat com a resultat una QoE molt satisfactòria per al consumidor, oferint-li un ecosistema complet de consum de continguts que inclou tant contingut multimèdia tradicional com contingut omnidireccional i mulsemedia. / [EN] Currently, there is a wide variety of multimedia content distribution technologies, consumer devices, as well as types of content. In terms of distribution technologies, such content can be delivered either over broadcast networks, such as DVB networks, or over broadband networks, such as IP networks (e.g. over the Internet). The coordination and convergence between the two types of technologies to distribute related content offers great potential to content providers, as well as great benefits to consumers (a proof of this is the Hybrid Broadcast Broadband TV or HbbTV standard). However, the reception of related content from different hybrid networks (broadcast/broadband) entails significant delay variability, so a number of important challenges must be overcome, such as the need for mechanisms to ensure synchronized consumption of such content, either on the same device or on different devices (local or remote). We have specified, implemented and evaluated synchronization solutions for hybrid content playback processes (broadcast and/or broadband networks) both in multi-device environments, between local devices (Inter-Device Synchronization or IDES), and in multi-user environments, between distributed network recipients (Inter-Destination Media Synchronization or IDMS), including the synchronization of multi-sensory effects. An end-to-end platform has been designed and implemented for the generation, distribution (adaptive) and consumption of related hybrid content, both on a single device and on multiple local (e.g., multiscreen scenario) or remote (e.g., Social TV) devices. Traditional (2D) and omnidirectional (360) multimedia content has been generated and prepared, enriched with multisensory effects that enhance the degree of realism and immersiveness for users. The platform is compatible with the HbbTV standard and, in addition, includes efficient solutions for key aspects that are not specified in that standard, but are necessary for the successful implementation of hybrid and multi-device TV services on it, such as: the need for a buffer for broadcast content (existing and optional up to the HbbTV 2. 0.2); mechanisms for signaling, association and description of content available and related to the main content; coordination and interaction mechanisms between consumption devices; or synchronization solutions to align in time the consumption of content in a satisfactory way. The results obtained in the evaluation carried out show that the level of synchronization acquired between the reproduction processes of the different types of content available through different transmission technologies reaches sufficient values to provide the user with a consumption with an imperceptible level of asynchronies. The accuracy achieved has resulted in a very satisfactory QoE for the consumer, offering him a complete content consumption ecosystem that includes traditional multimedia content as well as omnidirectional and mulsemedia content. / Marfil Reguero, D. (2022). Especificación, implementación y evaluación de soluciones de sincronización multimedia híbrida (broadcast/broadband) e inter-destinatario para posibilitar nuevas experiencias televisivas enriquecidas, personalizadas, inmersivas y compartidas [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/185746 / TESIS