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Survival and differentiation of central noradrenergic neurons /Holm, Pontus, January 2002 (has links)
Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 4 uppsatser.
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Reducing fibrosis and cell death in cardiomyoplasty /Robey, Thomas Edwin. January 2007 (has links)
Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 109-124).
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Tratamento de caldo e tipos de fermentos sobre os componentes secundários e qualidade da cachaça de alambique /Garcia, Graciany. January 2016 (has links)
Orientador: Márcia Justino Rossini Mutton / Resumo: Cachaça is the second most widely consumed alcoholic beverage in Brazil, obtained by distillation of sugarcane wine, having its quality affected by the raw material, fermentation conditions and productive process. With the growing demand of the consumer market for quality cachaça, the search for the improvement in relation to the progress of the technical-scientific knowledge has been intensified. The physical chemical treatment of the juice is a technology that qualifies the beverage. This study aimed is to evaluate the performance of two types of yeast (selected CA-11 and the pressed) and the influence of the physical-chemical treatment of sugarcane juice on the secondary compounds and the interference of the same in the quality of the beverage. The experiment was carried out in 2014/2015, using a variety of SP83-2847 cane grown in Pitangueiras-SP region. The trial design was done in blocks with 9 repetitions, 3 cycles with 3 repetitions, where the primary treatment was the clarified juice and not clarified and secondary the two types of yeast. It has assessed the viability of the cells and shoots and the index of budding along the fermentation. In the wine was determined the total acidity, pH, ARRT, alcoholic content, glycerol, efficiency fermentative and mineral composition. In distillates was analyzed total aldehydes, volatile acidity, total esters, methanol, acrolein, ethyl carbamate, furfuraldehyde and hydroxymethylfurfural, higher alcohols, alcohol sec. butyl and n-bu... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: A cachaça é a segunda bebida alcoólica mais consumida no Brasil, obtida pela destilação do vinho de cana-de-açúcar, tendo sua qualidade afetada pela matéria-prima, condições da fermentação e processo produtivo. Com a crescente exigência do mercado consumidor por cachaça de qualidade, intensificou-se a busca pelo aprimoramento em relação ao avanço do conhecimento técnico-científico. O tratamento físico químico do caldo é uma tecnologia que pode qualificar a bebida. Objetivou-se avaliar o desempenho de dois tipos de fermento (selecionado CA-11 e o prensado) e a influência do tratamento físico-químico do caldo de cana sobre os compostos secundários e a interferência dos mesmos na qualidade da bebida. O experimento foi realizado na safra 2014/2015, utilizando a variedade de cana SP83- 2847 cultivada na região de Pitangueiras-SP. O delineamento experimental utilizado foi feito em blocos com 9 repetições, sendo 3 ciclos com 3 repetições, onde o tratamento primário foi o caldo clarificado e não clarificado e o secundário os dois tipos de fermento. Avaliou-se a viabilidade das células e de brotos e o índice de brotamentos ao longo da fermentação. No vinho determinou-se acidez total, pH, ARRT, teor alcoólico, glicerol, eficiência fermentativa e composição mineral. Nos destilados foram analisados aldeídos totais, acidez volátil, ésteres totais, metanol, acroleína, carbamato de etila, furfural e hidroximetilfurfural, álcoois superiores, álcool sec. butílico e n-butanol, todos determinad... (Complete abstract click electronic access below) / Mestre
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Análise in vitro da N-Acetilcisteína em fibroblastos do ligamento periodontal estimulados por LPS bacteriano /Bittencourt, Tatiane Sampaio. January 2018 (has links)
Orientador: Carlos Henrique Ribeiro Camargo / Coorientadora: Profa. Dra. Gleyce Oliveira Silva / Banca : Eduardo Bresciani / Banca : Aleteia Massula de Melo Fernandes / Resumo: O objetivo deste estudo foi avaliar in vitro o comportamento biológico celular da N-Acetilcisteína (NAC), diante da estimulação ou não pelo LPS bacteriano. Foram utilizados fibroblastos do ligamento periodontal, que ficaram em contato por 48 horas com as substâncias testadas: NAC, Hidróxido de cálcio p.a. (HC), Lipopolissacarídeo de Escheria Coli (LPS), NAC + LPS e HC + LPS. Para os grupos NAC + LPS, HC + LPS e LPS, as células foram estimuladas com 2µg/mL de LPS por 24 horas, previamente aos tratamentos descritos. Foram realizados os testes biológicos de viabilidade celular (XTT), Espécies Reativas de Oxigênio (ROS), Elisa (para as citocinas IL-6, IL-8, IL-10, IL-1β e TNF-α) e Micronúcleo (MNT). Os dados foram analisados estatisticamente pela análise descritiva e pelos testes ANOVA e Kruskall Wallis, seguido do teste Dunn (p˂0.05) para os testes de XTT, ROS e MNT, e para o teste Elisa foi realizado cálculo das médias e desvio padrão. Os resultados obtidos mostraram que NAC teve um bom comportamento, frente à agressão provocada pelo LPS, quanto à produção de ROS. HC apresentou maior viabilidade celular que NAC, embora NAC não tenha apresentado citotoxicidade. Em relação à expressão das citocinas, NAC foi capaz de reduzir o potencial inflamatório do LPS quando da análise do TNF- α e IL-1β. NAC foi capaz de reduzir a genotoxicidade do LPS, como mostrado pelo teste MNT. Concluiu-se que NAC apresentou um comportamento biológico satisfatório, pela viabilidade celular dos fibroblast... (Resumo completo, clicar acesso eletrônico abaixo) / ABSTRACT The purpose of this study was to evaluate in vitro the action of N-Acetylcysteine (NAC) and calcium hydroxide (CH) on biological activity, either without or with LPS stimulation in periodontal ligament fibroblasts (PDLF) cells. PDLF were placed in contact with NAC, CH, NAC + LPS, LPS and CH + LPS for 48 hours. PDLF were stimulated by bacterial LPS for 24 hours in NAC + LPS, LPS and CH + LPS groups, before the substances described above are applied. The LPS and NAC effect on cell viability was measured using a XTT test. Reactive oxygen species (ROS) production was evaluated using ROS/superoxide detection kit. Inflammatory cytokines (IL-6, IL-8, IL-10, IL-1β and TNF-α) were evaluated by enzyme-linked immunosorbent (ELISA) assay. Genotoxicity was measured using micronucleus test (MNT). The means and standard deviation for all tests were calculated. Data were analyzed statistically by descriptive analysis, ANOVA and Kruskall Wallis tests, followed by Dunn test (p˂.05). CH was superior to NAC on cellular viability, although NAC was not cytotoxic. The results showed that NAC was able to reduce ROS production of LPS. NAC was able to reduce the inflammatory potential of LPS by decreasing the TNF-α and IL-1β release. NAC was able to reduce the genotoxicity of LPS. It was concluded that NAC showed a satisfactory biological activity, presenting a minimal effect on cell viability of PDLF cells and reducing ROS production and inflammatory potential provoked by LPS, and to decrease its genotoxicity / Mestre
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Resposta in vitro de células pré-osteoblásticas em cerâmica de hidroxiapatitaLuara Aline Pires 08 April 2015 (has links)
Com a evolução do Biomateriais houve melhorias nas opções de tratamentos e atualmente são utilizados em substituição de partes do corpo que foram perdidas e promovem a recuperação de funções biológicas. Dentre eles existem as chamadas Biocerâmicas, que incluem alumina, zircônia e derivadas de fosfato de cálcio. A hidroxiapatita possui composição e estrutura minerais semelhantes à fase mineral óssea e apresenta como propriedades a biocompatibilidade, osteocondutividade e bioatividade. O trabalho avaliou a viabilidade celular em cerâmica de hidroxiapatita experimental de origem bovina em comparação com dois tipos de zircônia comerciais e liga de titânio comercialmente puro, para que futuramente possa ser utilizada como material base para implantes dentários. A avaliação in vitro foi realizada por meio de testes nos quais células pré-osteoblásticas cultivadas de linhagem murina MC3T3-E1 foram colocadas em contato indireto e direto com estes materiais. Para viabilidade celular (n=8) foram feitos testes de ensaio MTT e Cristal Violeta em duplicata e após 24, 48 e 72 horas os níveis de absorbâncias foram analisados por meio de espectrofotometria no leitor de Elisa. Para as analises por microscopia eletrônica de varredura (n=6) as células foram plaqueadas diretamente sobre as superfícies dos discos, fixadas em vapor de tetróxido de ósmio 2% após 24 e 48 horas, seguido da metalização após 48 horas da fixação das células para análise em Microscópio Eletrônico de Varredura. Os resultados para viabilidade indireta foram submetidos ao teste paramétrico ANOVA, seguido de teste de Tukey (p<0,05). Tanto no teste de MTT quanto no Cristal Violeta, de acordo com o grupo controle positivo, todos os grupos apresentaram resultados satisfatórios. A cerâmica de hidroxiapatita não apresentou diferença estatística significante, demonstrando não ser um material citotóxico. Pelas imagens geradas no MEV do teste de viabilidade direta, verificou-se que houve adesão e proliferação das células nos dois períodos sobre as superfícies dos materiais. Portanto, pode-se afirmar que a cerâmica de hidroxiapatita apresentou-se como um material biocompatível. / With the evolution biomaterials there were improvements in treatment options, and are currently used in replacement body parts that were lost and promote the recovery of biological functions. Among them are the bioceramic which include alumina, zirconia and calcium phosphate derivative. Hydroxyapatite has mineral composition and structure similar to bone mineral phase and can be used as a biomaterial having biocompatibility, osteoconductivity and bioactivity. The study evaluated the cell viability in experimental hydroxyapatite ceramic bovine compared the two types of commercial zirconia and titanium alloy commercially pure, so that in future it can be used as base material for dental implants. In vitro evaluation was carried by means of tests in which the pre-osteoblastic cells MC3T3-E1 murine lineage cultured were placed in indirect and direct contact with these materials. For cell viability (n=8) were carried MTT assay and Crystal Violet tests in duplicate and after 24, 48 and 72 hours the absorbance levels were analyzed by spectrophotometry Elisa reader. For analysis by Scanning Electron Microscope variable pressure (n = 6) cells were plated directly on the discs surfaces, fixed in osmium tetroxide steam 2% after 24 and 48 hours, followed by metallization after 48 of cells fixation. The results for the cell viability were submitted to parametric test ANOVA, followed by Tukey test (p<0.05). Both in the MTT assay as Crystal Violet all groups exhibited satisfactory results absent cytotoxicity. By means of the SEM images produced, it was found that there was adhesion and proliferation of cells on the materials surfaces in the two periods. Therefore, it can be stated that the hydroxyapatite ceramic was presented as a biocompatible material.
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Dualité fonctionnelle de LMP1 : implication dans l’apoptose et la transformation cellulaire / Functionnal duality of LMP1 : involvement in apoptosis and cellular transformationBrocqueville, Guillaume 28 September 2011 (has links)
Le virus d’Epstein-Barr (EBV) est un herpèsvirus humain qui infecte plus de 90% de la population généralement de façon bénigne et asymptomatique. Cependant, de nombreuses données démontrent que ce virus peut également contribuer à certains processus de cancérisation. En effet, l’EBV est associé à de nombreuses pathologies malignes telles que le lymphome de Burkitt, le lymphome hodgkinien et le carcinome du nasopharynx. Dans la grande majorité de ces cancers associées à ce virus, l’EBV exprime un programme de latence de type II durant lequel la protéine LMP1 est exprimée. Elle est décrite comme l’oncogène majeur de l’EBV car son expression est nécessaire à la survie et à la prolifération des lignées transformées in vitro. Cette protéine membranaire est fonctionnellement apparentée aux membres de la famille des récepteurs du TNF. LMP1 est constitutivement active et son expression conduit à l’activation de voies de signalisation telles que les voies NF-κB, PI3K et des MAPK. L’activation de ces voies de signalisation cellulaire confère à LMP1 des propriétés oncogéniques, cependant, des effets toxiques liés à son expression ont également été décrits. Effectivement, LMP1 est capable d’induire l’apoptose dans différents types cellulaires. Dans ce contexte, nous avons d’abord développé et caractérisé, des variants dérivés de LMP1 constitués de sa partie C-terminale signalisatrice, complète ou partielle, fusionnée à la protéine GFP. Nous montrons que ces variants sont capables de séquestrer les protéines adaptatrices se fixant à LMP1 ou au récepteur TNFR1, et d’inhiber le signal et les phénotypes induits par ces derniers. Ces protéines à effet dominant négatif peuvent ainsi contrecarrer les effets transformants de LMP1 dans des modèles de latence II et III. Ces dominants négatifs peuvent aussi inhiber l’activation du TNFR1 et les phénotypes qui en découlent. Puis, nous avons étudié les propriétés de LMP1 en dehors d’un contexte infectieux et son rôle dans la transformation épithéliale. Nous démontrons que LMP1 induit la mort des cellules épithéliales MDCK mais certaines cellules outrepassent ses effets cytotoxiques générant des lignées qui expriment stablement LMP1 et dans lesquelles cet oncogène viral favorise la survie et exacerbe les phénotypes induits par le facteur de croissance HGF. Le caractère ambivalent de LMP1 pourrait limiter le pouvoir oncogène de l’EBV mais en contrepartie favoriser l’émergence de cellules résistantes à l’apoptose et capables de répondre de façon accrue à des facteurs de croissance. Nos travaux ont permis de mieux comprendre la dualité fonctionnelle de LMP1, d’une part ses effets oncogènes favorisant la survie cellulaire et d’autre part ses propriétés pro-apoptotiques, induites directement ou révélées suite à son inhibition, limitant la tumorigenèse. La caractérisation des mécanismes moléculaires impliquant LMP1 pourrait ainsi participer à la définition de potentielles stratégies thérapeutiques pour le traitement de cancers associés à l’EBV et où LMP1 est exprimée. / Epstein-Barr virus (EBV) is a human herpesvirus that infects more than 90% of worldwide population, generally asymptomatically. However, numerous studies show that EBV promotes tumorigenesis. Indeed, EBV infection is associated with many human malignancies including Burkitt’s lymphoma, Hodgkin’s lymphoma and nasopharyngeal carcinoma. In most of these cancers associated with EBV, it expresses latency II program in which the latent membrane protein 1 (LMP1) is expressed. LMP1 is described as the major EBV oncogene because its expression is necessary in vitro for survival and proliferation of transformed cell lines. This membrane protein is functionally related to members of the TNF receptors superfamily. LMP1 is constitutively active and its expression leads to activation of NF-κB, PI3K and MAPK signaling pathways. These activation confers oncogenic properties to LMP1, however, toxic effects associated with its expression are also described. Indeed, LMP1 can induce cell death in different cell types. In this context, we first developed and characterized LMP1 derivative variants consisting of its C-terminal signal, complete or partial, fused to GFP. We show that these variants are able to sequester adaptors binding to LMP1 and TNFR1, and inhibit signal and phenotypes induced by them. These proteins have dominant negative effect and may counteract LMP1 transformant properties in latency II cellular models. In addition, these dominant negatives impair TNFR1 signaling and associated phenotypes. Then, we studied LMP1 properties outside infectious context and its involvement in epithelial transformation. We show that LMP1 induces cell death in MDCK epithelial cells, but some go beyond its cytotoxic effects generating lines stably expressing LMP1 and in which this viral oncogene promotes survival and exacerbates HGF-induced phenotypes. Ambivalent character of LMP1 could limit the oncogenic potential of EBV but in return support the emergence of cells resistant to apoptosis and able to enhance growth factor responses. Our work allowed us to better understand the functional duality of LMP1 on the one hand its oncogenic effects favoring cell survival and other pro-apoptotic properties, induced directly or reveal by its inhibition, limiting tumorigenesis. Thus, characterization of molecular mechanisms involving LMP1 could participate in the definition of potential therapeutic strategies for treating cancers associated with EBV and where LMP1 is expressed.
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Expression of IGPR-1 in endothelial cells regulates cell survivalShafran, Jordan 03 November 2015 (has links)
Angiogenesis is a physiological process by which new blood vessels develop from preexisting vasculature. The process of converting endothelial cells into fully developed blood vessels involves multiple coordinated cellular events that occur through the collaboration that exists between a variety of growth factors, receptors and adhesion molecules. The immunoglobulin-containing and proline rich receptor-1 (IGPR-1) is an IgSF containing adhesion molecule that has been recently identified as a novel regulator of angiogenesis in vitro. In this study, we provide evidence that IGPR-1 promotes cell survival in porcine aortic endothelial cells (PAE) and plays a role in the inhibition of p38 MAPK in vitro. Deletion of the extracellular domain of IGPR-1 abolished IGPR-1’s ability to inhibit phosphorylation of p38 MAPK and promote the survival of endothelial cells. Likewise, mutation of serines 186 (A186-IGPR-1) and 220 (A220-IGPR-1) on the cytoplasmic domain of IGPR-1 was also found to reduce both the promotion of cell survival and inhibition of p38 MAPK. These findings suggest that both domains of IGPR-1 are important for endothelial cell survival and the activation p38 MAPK.
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Host Signaling Response to Adhesion of Bifidobacterium infantisGann, Reed N. 01 May 2010 (has links)
Investigations of the molecular binding partners of the probiotic bacterium Bifidobacterium longum subspecies infantis (B. infantis) and the pathogen Salmonella enterica subspecies enterica serovar Typhimurium LT2 (Salmonella ser. Typhimurium) found that these two very different bacteria bind gangliosides. However, these organisms lead to completely different host health outcomes when present in the gut. B. infantis is the founding microbial population in the intestinal tract of breast-fed infants. S. typhimurium is the most important food-borne pathogen that results in humans. This study used an in vitro gut epithelial cell model to examine the host cellular response to adhesion of B. infantis, which led to an increase in intestinal epithelium survival. This observation led to a series of experiments to elucidate the pathway for host signaling initiated by adherence of B. infantis to the host membrane to explain the increase in host cell survival. B. infantis adhesion induced significant (q≤0.05) differential expression of 208 host genes. These genes were associated with increased broad mechanisms of cell survival that included BIRC3, TNFAIP3, and SERPINB9. We hypothesized that a biochemical link existed between the host membrane adhesion protein and the increase in cell survival, mediated via AKT. We tested this hypothesis to demonstrate that B. infantis interaction initiated signal transduction using G-proteins via phosphorylation of AKT and induced production of the BIRC3, TNFAIP3, and SERPINB9. This study discovered adhesion of B. infantis initiated activation of AKT via phosphorylation of both Ser473 and Thr308, which results in increased cell survival.
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Histone Deacetylase 6 (HDAC6) Is Critical for Tumor Cell Survival and Promotes the Pro-Survival Activity of 14-3-3ζ viaDeacetylation of Lysines Within the14-3-3ζ Binding PocketMortenson, Jeffrey Benjamin 01 July 2015 (has links) (PDF)
Our understanding of non-histone acetylation as a means of cellular regulation is in its infancy. Using a mass spectrometry approach we identified acetylated lysine residues and monitored acetylation changes across the proteome as a consequence of metabolic stress (hypoxia). We observed changes in acetylation status of non-histone lysines in tumor cells. Through the use of small molecule inhibitors of histone deacetylase enzymes (HDACs) and siRNA screening identified HDAC6 as a pro-survival regulator of lysine acetylation during hypoxia. The phospho-binding protein 14-3-3ζ acts as a signaling hub controlling a network of interacting partners and oncogenic pathways. We show here that lysines within the 14-3-3ζ binding pocket and protein-protein interface can be modified by acetylation. The positive charge on two of these lysines, K49 and K120, is critical for coordinating 14-3-3ζ-phosphoprotein interactions. Through screening, we identified HDAC6 as the K49/K120 deacetylase. Inhibition of HDAC6 blocks 14-3-3ζ interactions with two well-described interacting partners, Bad and AS160, which triggers their dephosphorylation at S112 and T642, respectively. Expression of an acetylation-refractory K49R/K120R mutant of 14-3-3ζ rescues both the HDAC6 inhibitor-induced loss of interaction and S112/T642 phosphorylation. Furthermore, expression of the K49R/K120R mutant of 14-3-3ζ inhibits the cytotoxicity of HDAC6 inhibition. These data demonstrate a novel role for HDAC6 in controlling 14-3-3ζ binding activity.
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PurA sensitizes cells to toxicity induced by oxidative stressAlbukhaytan, Hawra, 0000-0003-1382-5645 January 2022 (has links)
PurA is an abundant and an evolutionary conserved protein that is known to bind to single stranded DNA or RNA and regulate both transcription and translation. PurA has been shown to be implicated in many neurological and neurodevelopmental deficits due to its multifunctional roles. In this study, we have studied the cells’ stress response in the presence and absence of PurA by introducing oxidative stress to the cells derived from wildtype (PurA+/+) mouse embryo fibroblasts (MEFs) and (PurA -/-) knockout mice. Our observations indicated that the presence of PurA is making cells more sensitive to toxicity compared to those cells lacking PurA, emphasizing on the importance of PurA in cell survival and cell cycle regulation. Our MTT results and western blot analysis were in agreement with each other confirming PurA ‘s importance in normal cell response against stress, in particular oxidative stress induced by Paraquat. PurA+/+ wildtype MEFs showed more sensitivity and a decrease in relative cell viability after 24 Hrs. exposure to 1.5 mM paraquat while less sensitivity was observed in PurA -/- knockout MEFs under same conditions. Western blot analysis showed a significantly increased expression of the apoptotic marker cleaved caspase3 in PurA+/+ wildtype MEFs under oxidative stress induction compared to PurA -/- knockout MEFs. Also, autophagy activation was measured by checking the expression of LC3-I and LC3-II in both MEFs under control and stressed conditions and the results were confirming that autophagy induction under oxidative stress is greater in the presence of PurA. PurA appears to be very critical for cell survival, and any alteration in its expression may lead to undesired consequences. / Biology
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