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A 12-month clinical trial examining the effects of a surface sealant on Class I composite resin restorations.Nahsan, Flavia Pardo Salata, Wang, Linda, Modena, Karin Silva, Francisconi Dos Rios, Luciana Fàvaro, Silva, Luciana Mendonça da, Calabria, Marcela Pagani, Casas-Apayco, Leslie, Mondelli, Rafael Francisco Lia 03 1900 (has links)
El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / A split-mouth, double-blind trial evaluated the effects of a surface sealant on the clinical performance of Class I composite resin restorations. In 16 patients, 27 pairs of maxillary and mandibular molars or premolars with Class I carious lesions or unsatisfactory restorations were restored with composite resin. For each pair, 1 surface was sealed with surface sealant. Clinical evaluations of marginal integrity, marginal discoloration, anatomical form, and secondary caries were performed by 2 experienced operators using modified US Public Health Service criteria 1-2 weeks and 6 and 12 months after treatment. Data were analyzed with the McNemar test (P < 0.05). After 6 months, only 1 (4%) sealed restoration presented a Bravo rating for marginal integrity. After 12 months, the Bravo ratings for marginal integrity were 2 (7%) for sealed restorations and 1 (4%) for nonsealed restorations. Restorations received a score of Alfa for all other parameters at all time periods. There were no statistically significant differences within or between the sealed and nonsealed groups (P = 1.0). The use of a surface sealant did not improve the clinical performance of posterior composite resin Class I restorations. / Revisión por pares
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Avaliação do tempo e do grau de eficiência do tratamento da má oclusão de classe I realizado com e sem extrações de pré-molares / Evaluation of time and efficiency of Class I malocclusion treatment carried out with and without premolar extractionsSalazar, Ruben Leon 20 January 2009 (has links)
O objetivo deste estudo retrospectivo foi comparar os resultados oclusais o tempo e a eficiencia do tratamento da ma oclusao de Classe I, realizado com e sem extracoes de pre-molares. Para tanto foi selecionada a partir das documentacoes do arquivo da Disciplina de Ortodontia da Faculdade de Odontologia de Bauru, uma amostra composta pelas documentacoes de 111 pacientes com ma oclussao de Classe I, e em seguida dividida em dois grupos que apresentaram as seguintes caracteristicas: Grupo 1, constituido por 65 pacientes (24 masculino e 41 feminino) com idade inicial media de 13,82 anos (minima de 10,69 e maxima de 22,04 anos), que foram tratados com extracoes de quatro pre-molares. Grupo 2, constituido por 46 pacientes, (16 masculino e 30 feminino) com idade inicial media de 14,01 anos (minima de 11,04 e maxima de 21,54 anos) tratados sem extracoes de pre-molares. Ambos os grupos foram tratados com aparelho fixo, utilizando a mecanica edgewise simplificada. As avaliacoes oclusais foram realizadas em modelos de gesso dos pacientes nas fases inicial e final utilizando o indice PAR. A avaliacao da compatibilidade no inicio do tratamento foi realizada por meio do teste do Qui- Quadrado e o teste t. As comparacoes entre os resultados oclusais, tempo e eficiencia do tratamento foram realizadas tambem por meio do test t e foi realizada a analise de regressao linear multipla para avaliacao das variaveis que poderiam estar relacionadas com o tempo de tratamento. Os resultados demonstraram que os grupos obtiveram resultados e alteracoes oclusais semelhantes, nao entanto, o tempo de tratamento foi menor e a eficiencia maior no grupo 2. Na analise de regressao linear multipla o modelo estatistico explicou 15% da variacao no tempo de tratamento, sendo que o protocolo de tratamento com extracoes foi a unica variável estatisticamente significante, a qual mostrou uma relacao direta com o tempo de tratamento. Pode-se concluir que, o tratamento com extracoes de pre-molares prove resultados oclusais e porcentagem de alteracoes oclusais semelhantes num maior tempo de tratamento, demonstrando por tanto um menor grau de eficiencia quando comparado ao tratamento realizado sem extracoes. / The objective of this retrospective study was to compare the occlusal outcomes, duration and efficiency of the treatment of Class I malocclusions carried out with and without premolar extractions. Accordingly, initial and final clinical charts and models of patients treated with fixed edgewise appliances were selected from the files of the Orthodontic Department at Bauru Dental School. Complete records of 111 patients were obtained and then divided into two groups: Group 1 consisted of 65 patients (24 male, 41 female), at an initial mean age of 13.82 years (range, 10.69 to 22.04 years) treated with four premolar extractions. Group 2 consisted of 46 patients (16 male; 30 female), at an initial mean age of 14.01 years (range, 11.04 to 21.54 years) treated without extractions. Initial and final occlusal evaluations were accomplished in study models of the patients using the peer assessment rating (PAR) index. Compatibility was evaluated with Chi-Square and t tests. The occlusal outcome, treatment duration and efficiency of the groups were also compared with t test and the variables that may be related to the duration of treatment were evaluated using the multiple linear regression analyses. Results showed that both groups had similar final occlusal outcomes and PAR reduction, however, Group 2 showed a significantly smaller treatment time and greater treatment efficiency index than group 1. In the regression analysis, the 9 selected variables explained 15% of the variance in treatment time and the extraction treatment protocol was the only statistically significant variable which was positively associated with treatment time. It can be concluded that premolar extractions influence significantly in treatment duration of Class I malocclusions and the use of this protocol provides the same occlusal outcome and occlusal improvement in a greater time and consequently less efficiency degree than non-extraction protocol.
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À procura do Ped bovino / Searching for the Ped genePuelker, Raquel Zaneti 30 March 2005 (has links)
O objetivo deste trabalho foi buscar genes MHC classe I em bovinos expressos durante o desenvolvimento embrionário e gestação que apresentassem similaridade ao gene Ped murino. Para tanto, embriões de PIV e placentas de fetos produzidos por monta natural ou clonados tiveram seu RNA extraído. A partir do RNA extraído foi realizada a produção de cDNA para tentar o isolamento de fragmentos com similaridade ao gene descrito em murinos. Uma vez isolados, os fragmentos foram purificados ou clonados em plasmídeo. As amostras foram seqüenciadas e as seqüências obtidas foram avaliadas, editadas e comparadas a outras seqüências pelo programa BLAST. A expressão de proteína MHC-Ib foi verificada em embriões (D7) e nas amostras de placenta utilizando anticorpo monoclonal de camundongo anti-Qa-2 marcado com FITC. A seqüência consenso obtida produziu alinhamentos significantes com os genes do complexo Bola que faz parte do MHC classe I bovino. O alinhamento entre a seqüência consenso e as seqüências de Q7 (Ped) e HLA-G publicados anteriormente demonstrou poucas variações de nucleotídeos entre as seqüências. As amplificações de cDNA de embriões que atingiram o quarto ciclo celular em até 48 hpi (R8) e em 48-90 hpi (L8) e de embriões que atingiram o estádio de blastocisto expandido em 7 dias de cultivo (RR) e em 9 dias de cultivo (RL) mostraram que o fragmento cuja seqüência apresenta alta similaridade ao gene Ped, está presente em embriões RR e RL, mas não em embriões no estádio de 8 células. Em placentas, o resultado da PCR mostrou a amplificação de oito fragmentos em diferentes fases da gestação. O sequenciamento dos fragmentos gerou nove seqüências consenso similares ao gene Ped e também a seqüências obtidas a partir de embriões bovinos. O alinhamento das seqüências mostrou a existência de duas isoformas contendo edição alternativa no exon 2 ou no exon 3. O resultado indicou a existência de polimorfismo ou mais de um gene com alta similaridade ao gene Q7 sendo transcritos durante o período gestacional. A expressão de proteína MHC-Ib foi verificada na membrana de células do trofoblasto e da MCI de embriões e na porção materno fetal de placentas, com uma maior expressão na porção fetal. Placentas oriundas de fetos clonados apresentam maior fluorescência comparada àquelas oriundas de monta natural. Este trabalho identificou, em embriões bovinos, um possível gene homólogo ao Ped murino e a expressão de proteína semelhante àquela codificada por este gene e conclui que existem vários transcritos com similaridade ao gene Q7 sendo expressos na região materno-fetal da placenta, durante a gestação em bovinos, assim como nos embriões / The aim of the present work was to identify bovine MHC class I genes similar to the murine Ped gene expressing during embryonic development and pregnancy. Bovine in vitro produced embryos and placentas obtained from natural mating or cloning-derived fetuses had their RNA extracted. cDNA was synthesized to isolate fragments showing similarity with the gene described in mice. Once isolated the fragments were purified or cloned into plasmids. The samples were sequenced and obtained sequences edited and compared to other sequences by means of the BLAST software. The expression of MHC-Ib protein was evaluated in embryos (D7) and in placentas using a FITC conjugated mouse monoclonal antibody anti-Qa-2. The consensus sequence amplified from blastocyst cDNA produced significant alignments with genes of the Bola complex, which is part of the bovine MHC class I. The alignment between the consensus sequence and the previously published Q7 and HLA-G sequences demonstrated little nucleotide variations among the sequences. The cDNA amplifications of embryos at the fourth cellular cicle in 48hpi (F8); 48-90hpi (S8) and embryos that reached blastocyst expanded stage in 7 days (FF); in 9 days (FS) showed that the Ped candidate gene is present in FF and FS but not in 8-cell embryos. In placentas, the PCR resulted in amplification of eight fragments at different pregnancy stages. Sequencing of the fragments generated nine consensus sequences, similar to the Ped gene and also to sequences obtained from bovine embryos. The alignment of the sequences showed the existence of two isoforms containing an alternative splicing on exon 2 or 3. This result indicates the existence of polymorphisms or that more than one gene with high similarity to the Q7 gene are transcribed during pregnancy. Expression of MHC-Ib protein was verified in trophoblast membrane and embryos inner cell mass cells and in the maternal-fetal portion in placentas, with a greater expression in the fetal portion. Cloned fetuses placentas presented grater fluorescence compared to natural mated. Overall this work identified in bovine embryos a possible gene homologous to the murine Ped and the expression of protein similar to that coded by this gene and concluded that there are many transcripts similar to the Q7 gene being expressed in the maternal-fetal region of the placenta during pregnancy in bovine and such as in embryos
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Avaliação da metodologia de cálculo de dose em microdosimetria com fontes de elétrons com o uso de código MCNP5 / Evaluation of the methodology for dose calculation in microdosimetry with electrons sources using the MCNP5 codeCintra, Felipe Belonsi de 26 November 2010 (has links)
Este trabalho realizou uma comparação entre alguns dos principais códigos de transporte que empregam a abordagem estocástica de Monte Carlo para aplicação em cálculos dosimétricos em Medicina Nuclear. Foram analisados com detalhes os diversos modelos físicos e numéricos utilizados pelo código MCNP5 em relação códigos como Penelope e EGS. A identificação de suas potencialidades e limitações para solução de problemas microdosimétricos foram destacados. A metodologia condensada usada pelo MCNP resultou em valores para energia depositada normalmente menores, evidenciando uma conhecida característica do método das historias condensadas: o fato de subestimar tanto o número de colisões ao longo da trajetória do elétron quanto do número de partículas secundárias criadas. O uso de códigos de transporte como Penelope e MCNP em escalas micrométricas recebeu especial atenção neste trabalho. Códigos classe I e II foram estudados e seus principais recursos foram explorados visando o transporte de elétrons, que são de especial importância em dosimetria. Espera-se que a avaliação das metodologias disponíveis, aqui abordadas contribua para um maior entendimento do comportamento de tais códigos principalmente para esta classe de problemas, comuns em microdosimetria. / This study made a comparison between some of the major transport codes that employ the Monte Carlo stochastic approach in dosimetric calculations in nuclear medicine. We analyzed in detail the various physical and numerical models used by MCNP5 code in relation with codes like EGS and Penelope. The identification of its potential and limitations for solving microdosimetry problems were highlighted. The condensed history methodology used by MCNP resulted in lower values for energy deposition calculation. This showed a known feature of the condensed stories: its underestimates both the number of collisions along the trajectory of the electron and the number of secondary particles created. The use of transport codes like MCNP and Penelope for micrometer scales received special attention in this work. Class I and class II codes were studied and their main resources were exploited in order to transport electrons, which have particular importance in dosimetry. It is expected that the evaluation of available methodologies mentioned here contribute to a better understanding of the behavior of these codes, especially for this class of problems, common in microdosimetry.
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À procura do Ped bovino / Searching for the Ped geneRaquel Zaneti Puelker 30 March 2005 (has links)
O objetivo deste trabalho foi buscar genes MHC classe I em bovinos expressos durante o desenvolvimento embrionário e gestação que apresentassem similaridade ao gene Ped murino. Para tanto, embriões de PIV e placentas de fetos produzidos por monta natural ou clonados tiveram seu RNA extraído. A partir do RNA extraído foi realizada a produção de cDNA para tentar o isolamento de fragmentos com similaridade ao gene descrito em murinos. Uma vez isolados, os fragmentos foram purificados ou clonados em plasmídeo. As amostras foram seqüenciadas e as seqüências obtidas foram avaliadas, editadas e comparadas a outras seqüências pelo programa BLAST. A expressão de proteína MHC-Ib foi verificada em embriões (D7) e nas amostras de placenta utilizando anticorpo monoclonal de camundongo anti-Qa-2 marcado com FITC. A seqüência consenso obtida produziu alinhamentos significantes com os genes do complexo Bola que faz parte do MHC classe I bovino. O alinhamento entre a seqüência consenso e as seqüências de Q7 (Ped) e HLA-G publicados anteriormente demonstrou poucas variações de nucleotídeos entre as seqüências. As amplificações de cDNA de embriões que atingiram o quarto ciclo celular em até 48 hpi (R8) e em 48-90 hpi (L8) e de embriões que atingiram o estádio de blastocisto expandido em 7 dias de cultivo (RR) e em 9 dias de cultivo (RL) mostraram que o fragmento cuja seqüência apresenta alta similaridade ao gene Ped, está presente em embriões RR e RL, mas não em embriões no estádio de 8 células. Em placentas, o resultado da PCR mostrou a amplificação de oito fragmentos em diferentes fases da gestação. O sequenciamento dos fragmentos gerou nove seqüências consenso similares ao gene Ped e também a seqüências obtidas a partir de embriões bovinos. O alinhamento das seqüências mostrou a existência de duas isoformas contendo edição alternativa no exon 2 ou no exon 3. O resultado indicou a existência de polimorfismo ou mais de um gene com alta similaridade ao gene Q7 sendo transcritos durante o período gestacional. A expressão de proteína MHC-Ib foi verificada na membrana de células do trofoblasto e da MCI de embriões e na porção materno fetal de placentas, com uma maior expressão na porção fetal. Placentas oriundas de fetos clonados apresentam maior fluorescência comparada àquelas oriundas de monta natural. Este trabalho identificou, em embriões bovinos, um possível gene homólogo ao Ped murino e a expressão de proteína semelhante àquela codificada por este gene e conclui que existem vários transcritos com similaridade ao gene Q7 sendo expressos na região materno-fetal da placenta, durante a gestação em bovinos, assim como nos embriões / The aim of the present work was to identify bovine MHC class I genes similar to the murine Ped gene expressing during embryonic development and pregnancy. Bovine in vitro produced embryos and placentas obtained from natural mating or cloning-derived fetuses had their RNA extracted. cDNA was synthesized to isolate fragments showing similarity with the gene described in mice. Once isolated the fragments were purified or cloned into plasmids. The samples were sequenced and obtained sequences edited and compared to other sequences by means of the BLAST software. The expression of MHC-Ib protein was evaluated in embryos (D7) and in placentas using a FITC conjugated mouse monoclonal antibody anti-Qa-2. The consensus sequence amplified from blastocyst cDNA produced significant alignments with genes of the Bola complex, which is part of the bovine MHC class I. The alignment between the consensus sequence and the previously published Q7 and HLA-G sequences demonstrated little nucleotide variations among the sequences. The cDNA amplifications of embryos at the fourth cellular cicle in 48hpi (F8); 48-90hpi (S8) and embryos that reached blastocyst expanded stage in 7 days (FF); in 9 days (FS) showed that the Ped candidate gene is present in FF and FS but not in 8-cell embryos. In placentas, the PCR resulted in amplification of eight fragments at different pregnancy stages. Sequencing of the fragments generated nine consensus sequences, similar to the Ped gene and also to sequences obtained from bovine embryos. The alignment of the sequences showed the existence of two isoforms containing an alternative splicing on exon 2 or 3. This result indicates the existence of polymorphisms or that more than one gene with high similarity to the Q7 gene are transcribed during pregnancy. Expression of MHC-Ib protein was verified in trophoblast membrane and embryos inner cell mass cells and in the maternal-fetal portion in placentas, with a greater expression in the fetal portion. Cloned fetuses placentas presented grater fluorescence compared to natural mated. Overall this work identified in bovine embryos a possible gene homologous to the murine Ped and the expression of protein similar to that coded by this gene and concluded that there are many transcripts similar to the Q7 gene being expressed in the maternal-fetal region of the placenta during pregnancy in bovine and such as in embryos
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Immune recognition molecules in synaptic plasticity and regeneration of spinal motoneuronsThams, Sebastian, January 2009 (has links)
Diss. (sammanfattning) Stockholm : Karolinska institutet, 2009.
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Immunological assays relevant to definition of bovine theileria parva-specific cytotoxic CD8+ T cell responsesMusembi, Susan Mbithe January 2012 (has links)
A major objective in Theileria parva subunit vaccine development is to induce a vaccine antigen specific response mediated by cytotoxic CD8+ T cells (CTL). Therefore it is essential to be able to measure the frequency of the responding CD8+ T cells after vaccination and correlate it with a clinical outcome on challenge. Recently concluded immunogenicity and efficacy studies of T. parva specific CTL antigens showed successful induction of CTL responses in some animals, which correlated with reduced disease severity after challenge. To provide correlates of immunity antigen-specific CD8+ T cell mediated IFN-γ responses and CTL lytic responses were measured over the course of the experiments. Several challenges presented in these trials aimed at optimising vaccine efficacy. While the IFN-γ ELISPOT is a sensitive and reliable assay widely used in vaccine research, the use of chromium/indium release assay remains to be the only assay in use that measures T. parva-specific CTL activity. Hence the overall goal of the study was to develop novel reagents and novel assays to identify parasite-specific CD8+ T lymphocytes with lytic potential. To address this objective, bovine perforin, granzymes A and B, as specific effector proteins expressed in activated CTL were cloned and expressed using a baculovirus expression system. Sequence analysis of the cloned cDNAs showed the isolated cDNA belonged to the perforin and granzyme sub-families respectively. Perforin cDNA demonstrated 85% homology to human perforin with presence of conserved regions resembling calcium binding motif, membrane attack complex component as well complement protein. The sequences encoded by the cloned granzyme A and B cDNAs have the features of a trypsin like serine protease and demonstrates over 70% homology to the human cDNA over the active enzyme region as well catalytic residues characteristic of serine proteases. The expressed polypeptides of all three proteins were used to produce specific antibodies for use as reagents in immunoassays including ELISpot and intracellular staining for flow cytometric analysis. While the antibodies showed reactivity to the recombinant proteins, these reagents displayed different functionality in the recognition of the native protein. Peptide-major histocompatibility complexes (MHC) class I tetrameric complexes (tetramers) are proving invaluable as fluorescent reagents for enumeration, characterisation and isolation of peptide-specific CD8+ T cells and have afforded advantages to phenotype antigen-specific T cells with minimal in vitro manipulation. Fluorescent bovine tetramers were shown to specifically stain antigen-specific CTL by directly binding the T cell receptor (TCR). Analyses of CD8 T-cell responses in live-vaccine immunised cattle also showed that this method is robust and demonstrates changes in the kinetics and specificity of the CD8+ T cell response in primary and secondary infections with T. parva. On average, results of functional assays and tetramer staining followed parallel trends, measured roughly the same populations and allowed for surface and intracellular staining for CD8 T cell marker and perforin, respectively, demonstrating a method that reliably quantifies the frequency, phenotype and function of specific CD8+ T cells. The technical simplicity, rapidity and ability of the flow cytometric technique described in this thesis to measure low frequency antigen-specific responses suggests that tetramer staining, combined with functional assays could be broadly applicable to the valuation of vaccination efficacy to determine which protocols are most successful in inducing CTL responses.
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Rhesus macaque KIR recognition of MHC class I molecules: Ligand identification and modulation of interaction by SIV peptidesSchafer, Jamie Lynn 04 June 2015 (has links)
Natural killer (NK) cells can kill virus-infected cells without prior antigenic exposure, and are therefore important for controlling viral replication prior to the onset of adaptive immune responses. Primate NK cells express activating and inhibitory killer-cell immunoglobulin-like receptors (KIRs) that bind to specific major histocompatibility complex (MHC) class I molecules. The importance of KIR interactions with MHC class I in human immunodeficiency virus (HIV) pathogenesis is demonstrated by the association of select KIR and MHC class I genotypes with delayed progression to acquired immunodeficiency syndrome (AIDS).
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A Functional Study of the Major Histocompatibility Class I Antigen Presentation Pathway in Rainbow Trout (Oncorhynchus mykiss)Sever, Lital January 2014 (has links)
Major Histocompatibility Complex (MHC) class I receptors are glycoproteins which play a critical role in anti-viral immunity by displaying foreign peptides to cytotoxic T cell lymphocytes. The loading of high affinity peptides into the MHC class I receptor in mammals is coordinated by a multiple proteins that are collectively referred to as the peptide loading complex (PLC). To date, the composition of the peptide loading complex in fish is unknown and therefore the characterization of the molecules which may exist in this putative complex was pursued.
This thesis includes the cloning and functional characterization of ERp57 and calnexin in rainbow trout which, in mammals, are known to interact with the MHC class I receptor either during its early biogenesis or later in the assembly of the PLC. Trout ERp57 and calnexin cDNA sequences are ubiquitously expressed in trout tissues and both the ERp57 and calnexin genes appear in at least two copies each in the trout genome. Interestingly, despite their high sequence identity with their mammalian homologues, some structural discrepancies were identified. ERp57 does not contain an endoplasmic reticulum (ER) retention signal or a nuclear localization signal, while one of the two isolated cDNA clones for calnexin does not contain an ER (endoplasmic reticulum) retention signal and lacks a conserved C-terminal serine phosphorylation site. These findings suggest that in trout, there may be unique versions of these proteins that have acquired different cellular functions. Through the production of polyclonal antibodies against trout ERp57, the conserved protein induction of ERp57 during ER stress was demonstrated concurrently with calnexin.
In addition, this study shows for the first time that ERp57 can be induced transcriptionally by phytohemagglutinin and synthetic double stranded RNA, which implies its possible regulatory role during viral infection and the activation of the immune response. Furthermore, the functional characterization of the MHC class I specific chaperone tapasin, a key element in the PLC of mammals was pursued. Tissue and cell line distribution revealed that tapasin is expressed in high levels in immune system organs and in the rainbow trout macrophage cell line RTS11, at a relative molecular weight of 48 kDa with an additional 20 kDa band detected by the tapasin antibody. Tapasin protein was significantly up regulated upon exposure to synthetic double stranded RNA and during infection with two fish viruses: chum salmon virus and viral hemorrhagic septicemia virus genotype IVa, whereas the expression of the 20 kDa band was not affected by these stimuli.
This study also examined the regulation of the MH class I heavy chain,β2 microglobulin and their associated machinery upon exposure to viral hemorrhagic septicemia virus genotype IVa at permissive and non-permissive temperatures. β2 microglobulin secretion into the cell media, a marker of MH class I receptor turnover, was detected in the conditioned media of RTS11 cells under normal conditions and was shown to be significantly enhanced during viral hemorrhagic septicemia virus genotype IVa infection. Furthermore, when RTS11 cells were maintained at cold temperatures, the secretion of β2 microglobulin was significantly reduced in both infected and non-infected cultures, while the cellular levels of β2 microglobulin remained unchanged. These results suggest that cold temperature can alter the expression of the MH class I molecule on the cell surface and therefore may be contributing to host susceptibility to viral hemorrhagic septicemia virus genotype IVa during the winter.
Lastly, Co-immunoprecipitation demonstrated the interaction of the lectin chaperones: calnexin and calreticulin with the glycosylated MH class I receptor supporting their conserved role during MH class I receptor folding in fish. Concurrently, tapasin's interaction with transporter associated with antigen processing (TAP) and with the glycosylated form of the MH class I was revealed for the first time in fish, which supports their role in antigen presentation as in mammals. This study demonstrated that ERp57 and tapasin form a conserved disulfide linked heterodimer of 110 kDa, however unlike mammals, an additional 75 kDa heterodimer was detected which suggests a possible novel interaction of ERp57 with a 20 kDa tapasin version alternately regulating antigen presentation in fish.
Overall, this study suggest that the interactions involved in antigen presentation in mammals are conserved in fish, however the presence of different protein versions of calnexin, ERp57 and tapasin might dictate a different mode of regulation for MH class I assembly in fish, as opposed
to mammals. Elucidating these interactions during various viral infections in fish can help to uncover possible viral strategies to manipulate the host immune response and will provide information needed to assist in designing novel tools to prevent fish viral diseases.
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Premature Translational Termination and the Rapidly Degraded Polypeptide PathwayLacsina, Joshua Rene January 2012 (has links)
<p>Nearly thirty percent of all newly synthesized polypeptides are targeted for rapid proteasome-mediated degradation. These rapidly degraded polypeptides (RDPs) are the primary source of antigenic substrates for the major histocompatibility complex (MHC) class I presentation pathway, allowing for the immunosurveillance of newly synthesized proteins by cytotoxic T lymphocytes. Despite the recognized role of RDPs in MHC class I presentation, it remains unclear what molecular characteristics distinguish RDPs from their more stable counterparts. It has been proposed that premature translational termination products may constitute a form of RDP; indeed, in prokaryotes translational drop-off products are normal by-products of protein synthesis and are subsequently rapidly degraded. </p><p>To study the cellular fate of premature termination products, the antibiotic puromycin was used to modulate prematurely terminated polypeptide production in human cells. At low concentrations, puromycin doubled the fraction of rapidly degraded polypeptides, with enhanced degradation predominantly affecting small polypeptides, consistent with rapid degradation of truncated translation products. Immunoprecipitation experiments using anti-puromycin antisera demonstrated that the majority of peptidyl-puromycins are rapidly degraded in a proteasome-dependent manner. Low concentrations of puromycin increased the recovery of cell surface MHC class I-peptide complexes, indicating that prematurely terminated polypeptides can be processed for presentation via the MHC I pathway. In the continued presence of puromycin, MHC I export to the cell surface was inhibited, coincident with the accumulation of polyubiquitinated proteins. The time- and dose-dependent effects of puromycin suggest that the pool of peptidyl-puromycin adducts differ in their targeting to various proteolytic pathways which, in turn, differ in the efficiency with which they access the MHC class I presentation machinery. These studies highlight the diversity of cellular proteolytic pathways necessary for the metabolism and immunosurveillance of prematurely terminated polypeptides which are, by their nature, highly heterogeneous.</p> / Dissertation
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