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INFLUENCE OF HYDROSTATIC PRESSURE AND TEMPERATURE ON THE METABOLIC ACTIVITY OF Alcanivorax marisrubri sp. nov. ISOLATED FROM THE RED SEADelgadillo Ordoñez , Nathalia Catalina 03 1900 (has links)
Hydrostatic pressure (HP) and low temperature are among the main parameters that affect the microbial activity in the deep sea. Especially in the event of an oil spill, the natural microbial degradation of hydrocarbons in the harsh conditions of the deep sea can be significantly impaired. In the Red Sea, the temperature in the deep (22°C) is much higher than in other oceans and may favor hydrocarbon degradation. Bacteria of the genus Alcanivorax, which are prominent and ubiquitous alkane degrading bacteria, have been extensively studied because of their high abundance in oil-contaminated shallow water, but have been shown to be absent in the deep sea because of their piezo-sensitivity. In the present thesis, the novel species Alcanivorax marisrubri isolated at 1000 m from the Southern Red Sea has been evaluated for its piezo-adaptation under different combinations of temperature, and HP. A. marisrubri showed a piezotolerance different from other Alcanivorax species. Furthermore, a positive compensation of growth inhibition was observed when the cells were exposed to mild HP (10 MPa) in combination with a relatively high temperature of 38°C. While growth was inhibited at lower temperatures (20 and 26°C) under mild-HP (5 and 10 MPa), the metabolic activity was triggered, possibly in response to cellular stress. This study showed that the growth and metabolic activity of A. marisrubri under HP depend on temperature, which exerts a positive compensation effect and may extend the growth of this bacterium to the depths of the Red Sea.
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Accelerated Life Testing Of Subsea Equipment Under Hydrostatic PressureThiraviam, Amar Raja 01 January 2010 (has links)
Accelerated Life Testing (ALT) is an effective method of demonstrating and improving product reliability in applications where the products are expected to perform for a long period of time. ALT accelerates a given failure mode by testing at amplified stress level(s) in excess of operational limits. Statistical analysis (parameter estimation) is then performed on the data, based on an acceleration model to make life predictions at use level. The acceleration model thus forms the basis of accelerated life testing methodology. Well established accelerated models such as the Arrhenius model and the Inverse Power Law (IPL) model exist for key stresses such as temperature and voltage. But there are other stresses like subsea pressure, where there is no clear model of choice. This research proposes a pressure-life (acceleration) model for the first time for life prediction under subsea pressure for key mechanical/physical failure mechanisms. Three independent accelerated tests were conducted and their results analyzed to identify the best model for the pressure-life relationship. The testing included material tests in standard coupons to investigate the effect of subsea pressure on key physical, mechanical, and electrical properties. Tests were also conducted at the component level on critical components that function as a pressure barrier. By comparing the likelihood values of multiple reasonable candidate models for the individual tests, the exponential model was identified as a good model for the pressure-life relationship. In addition to consistently providing good fit among the three tests, the exponential model was also consistent with field data (validation with over 10 years of field data) and demonstrated several characteristics that enable robust life predictions in a variety iv of scenarios. In addition the research also used the process of Bayesian analysis to incorporate prior information from field and test data to bolster the results and increase the confidence in the predictions from the proposed model.
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The use of enzyme inhibitor and high hydrostatic pressure to formulate fish gels of superior qualitySareevoravitkul, Ramon January 1995 (has links)
No description available.
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Influence of layer waviness on the hydrostatic response of thick composite cylindersBrown, Timothy L. 19 September 2009 (has links)
The influence of layer waviness in thick cross-ply composite cylinders subjected to hydrostatic pressure is investigated. The cylinders considered are graphite-epoxy with a 2: 1 ratio of circumferential to axial layers. All cylinders considered contain 104 total layers with a layup of [90/(90/0/90h71s, where a '0° 1 layer is taken to be in the axial direction. The influence of a single isolated group of wavy layers in an otherwise perfect cylinder is evaluated. Layer waviness in only the circumferential direction is considered, and the analysis is assumed to be valid only away from the cylinder ends. A parametric investigation is performed to determine the combined influence of wave location, wave amplitude, and cylinder geometry on hydrostatic response of the cylinder, particularly the stresses generated in and around the wave. The wave is assumed to be located either at the inner or the outer radius of the cylinder. Three wave amplitudes, 0, are considered: 1/2, 1, and 2 layer thicknesses. Only waves with a half wave length of 10 layer thicknesses are considered. Three cylinder geometries are considered, specifically ones with radius to thickness ratios of 5, 10, and 20.
Finite element analysis is used to determine the stress state within the imperfect, i.e., wave included, cylinders. Based on a maximum stress failure criterion, failure pressures are determined for each of the various wave and cylinder geometries. Failure pressures for the imperfect cylinders are compared with those for a perfect cylinder to determine the failure pressure reduction ratios due to fiber waviness. It is shown that pressure capacity reductions of approximately 50% are possible for the range of parameters studied. Failure is primarily due to fiber compression, though interlaminar shear and interlaminar tension are a factor. Finite element analysis is also used to deter ine the failure pressure of the perfect cylinder due to buckling. This is done to determine whether failure due to buckling may overshadow material failure due to fiber waviness. It is shown that buckling is a factor in only one of the cylinder geometries considered, and only in the cases of mild layer waviness.
In addition to results, details about the finite element model are presented. These details include geometry of the wave, changes in material properties due to local fiber rotation and local volume fraction changes, boundary conditions, and justifications for modeling simplifications that were made in an effort to reduce computational costs and analysis times. / Master of Science
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Hurdle Technologies Using Essential Oils And High Hydrostatic Pressure To Inactivate E. Coli In Fresh BeefSahmurat, Fatma 08 December 2016 (has links)
In this study, potential of high hydrostatic pressure (HHP) and essential oils (EOs) as natural antimicrobials was evaluated to produce E. coli safe and quality beef product. First, the individual and combined effects of antimicrobial activity (minimum inhibitory concentration) of basil, black cumin, cilantro, cumin, fenugreek, ginger, oregano, black pepper, rosemary, thyme, turmeric oil emulsions on E. coli ATCC 25922 with and without HHP treatment were evaluated. Cumin, oregano and thyme EOs showed highest antimicrobial activity against E. coli ATCC 25922. The synergy of selected EOs against E. coli ATCC 25922 was determined using the checkerboard method to obtain fractional inhibitory concentration index. Although their combinations did not show synergy, they expressed synergy when combined with HHP (400 MPa, 10 min, 20 °C) and the best combination was cumin and oregano EOs with HHP.
Effects of HHP and EO combinations on inactivation of E. coli ATCC 25922 in beef were investigated using response surface methodology (RSM). Statistical analysis showed the model was significant for predicting log reduction with high accuracy. The significant model terms were pressure and time. Compared to control, HHP/EO treated samples showed no-post growth when stored up to 120 days at 4°C. Presented results suggests that the combination of HHP and antimicrobials has not only improved the process parameters (lowered pressure, time, and EO concentration) but also prevented recovery of E. coli ATCC 25922 during storage.
RSM was employed to analyze the synergistic effects of HHP and EOs on beef quality (color, texture and lipid oxidation). Color indices were significantly affected by pressure, time and their interactions. Above 400 MPa the discoloration was similar to cooked beef and EO addition did not help color improvement. However, EOs showed significant antioxidant activity on both treated and untreated samples during storage.
In conclusion, there is a great potential of HHP and EO combinations to enhance pathogen inactivation while keeping the quality of beef. Moreover, presence of EOs can prolong the shelf life of pressure treated beef. Therefore, the combination of HHP and EO is very promising for meat industry. / Ph. D. / Meat is a natural source of protein, essential vitamins, which makes it a nutrient-rich source of a healthy diet as well as an ideal environment for food-borne pathogens and spoilage bacteria. It is therefore essential to preserve very perishable meat products in terms of microbial contamination. As an alternative to many preservation methods such as chilling, canning, curing, smoking, dehydrating and heat treatment, a non-thermal mild food preservation technology of high hydrostatic pressure processing (HHP) is proposed for inactivating the most common meat contaminant bacteria of <i>E. coli</i>. Essential oils (EOs) can provide a solution for pasteurization requirements and reducing quality losses associated with HHP treatment.
In this study the synergistic effect of selected EOs (basil, black cumin, cilantro, cumin, fenugreek, ginger, oregano, black pepper, rosemary, thyme, turmeric oil emulsions) and HHP technology on inactivation of <i>E. coli</i> ATCC 25922 on contaminated meat cuts were investigated. Experimental design and statistical analysis were conducted using response surface methodology (RSM). Combination of HHP/EO treated samples showed no-post growth of <i>E. coli</i> ATCC 25922 when meat samples were stored up to 120 days at 4°C. Presented results are suggesting that HHP in combination with EOs has increased the log reduction of <i>E.coli</i> and as well, decreased the quality losses (color, lipid oxidation textural analysis) compared to control samples where HHP is applied alone. As a conclusion, this study shows that there is a great potential of HHP and EO combinations to enhance pathogen inactivation while keeping the quality of beef.
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Metamagnetismo no Ni(NO3)2.2H2O e sua dependência com pressão hidrostática / Metamagnetism in Ni(NO3)2.2H2O and its dependence hydrostatic pressureSugui Junior, Said Salem 21 September 1983 (has links)
A partir de medidas isotérmicas e isobáricas de susceptibilidade magnética diferencial, determinamos o diagrama de fases magnéticas do metamagneto Ni(NO3)2.2H2O, para pressões hidrostáticas de até cerca de 11 kbar. Determinamos o campo de transição à temperatura nula Hc(0,P) e à temperatura de Néel, ambos apresentando um crescimento aproximadamente linear com P. A partir de efeitos de desmagnetização, pudemos determinar o ponto de união das linhas de primeira e segunda ordem de cada diagrama, e para P=0 pudemos caracterizá-lo com um ponto tricrítico, com temperatura tricrítica Tt=3.70K. Observamos ainda a existência de uma transição metamagnética induzida por pressão para valores do intervalo de 0 a 8 kbar. Usando a Aproximação de Campo Médio num modelo elástico aproximado para metamagnetos com S=1, pudemos obter expressões para TN(P) e Hc(0,P), ambas apresentando uma dependência linear em P, em bom acordo com os dados experimentais. Usando este modelo obtivemos também uma expressão analítica para Tt(P). Uma vez que TN(P), Hc(0,P) e Tt(P) dependem dos parâmetros de \"exchange\", pudemos determinar a dependência de tais parâmetros com a pressão. Também através deste modelo simples, pudemos compreender que a transição metamagnética induzida por pressão está provavelmente associada a uma forte dependência do campo cristalino com a pressão / From isothermical and isobaric differential magnetic susceptibility measurements we have determined the magnetic phase diagram of the metamagnetic system Ni(NO3)2.2H2O, for several hydrostatic pressures up to 11 kbar. The zero temperature transition field Hc(0,P) and the Néel temperature TN(P) were determined, both increasing almost linearly with P. From demagnetizing effects we were able to determine the joint point of the first and second order lines of each diagram, which for P=0 is a tricritical point, with tricritical temperature Tt=3.70K. We also observed a pressure induced metamagnetic transition in the range from 0 to 8 kbar. Using Mean Field Theory in an approximated elastic S=1 metamagnetic model, we could obtain expressions for TN(P) and Hc(0,P), both showing linear dependence on P, in agreement with experimental data. We could also obtain, using this model, an analytical expression for Tt(P). Once TN(P), Hc(0,P) and Tt(P) are dependent on the exchange parameters, we cou1d obtain the pressure dependence of these parameters. From this simple model we could understand that the pressure induced metamagnetic transition is probably associated with a strong variation of the crystalline field with pressure
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Estudos de renaturação de proteínas agregadas utilizando altas pressões hidrostáticas / Renaturation studies of aggregate proteins using high hydrostatic pressureNatália Malavasi Vallejo 05 March 2013 (has links)
No presente trabalho estudamos a renaturação sob alta pressão hidrostática de uma forma mutante da proteína verde fluorescente (enhanced GFP, eGFP), a qual somente emite fluorescência característica quando enovelada na sua forma nativa. A abordagem do presente estudo foi focada no controle da bioatividade da proteína recombinante, a fluorescência, como alternativa à determinação de solubilidade da proteína, fator que não é um indicador ideal de enovelamento proteico adequado. A ação da alta pressão na solubilização dos corpos de inclusão (CI) de eGFP produzidos em bactérias E. coli recombinantes e no enovelamento da proteína foi estudada. A compressão dos CI de eGFP em 2,4 kbar durante 30 minutos promoveu a dissociação dos agregados. No entanto, a incubação nesta condição não favoreceu o enovelamento da eGFP. O processo de renaturação foi avaliado em diversas condições de descompressão após a dissociação em 2,4 kbar. Durante a descompressão gradual, o aumento da fluorescência foi obtido em pressões que variaram entre a pressão atmosférica e 1,38kbar. Os níveis mais elevados de fluorescência de eGFP foram obtidos por incubação durante várias horas a níveis de pressão entre 0,35 e 0,69 kbar. Esta condição de pressão se mostrou favorável à renaturação de eGFP e é possível que também possa ser utilizada para favorecer o enovelamento de outras proteínas monoméricas. Ainda utilizando a eGFP como modelo, verificamos que os CI desta proteína produzidos por bactérias cultivadas em menor temperatura (37ºC) possuem maior quantidade de proteína recombinante apresentando a fluorescência característica em 509 nm, ou seja, na sua forma nativa, do que os CI expressos em temperaturas mais elevadas (42ºC e 47ºC). A análise realizada por espectroscopia de infravermelho (FT-IR) também demonstrou que os CI produzidos em temperaturas mais brandas possuem maior grau de estruturas secundárias semelhantes às da proteína na sua forma nativa. Além disso, os CI produzidos a 37ºC também são mais facilmente solubilizados pela ação da alta pressão do que aqueles produzidos em maior temperatura. Conforme esperado, a renaturação da eGFP a partir de CI produzidos a 37ºC foi 25 vezes mais eficiente do que a obtida utilizando CI produzidos a 47ºC. No presente estudo demonstramos também que a dissociação dos agregados exercida pela ação da alta pressão (2,4 kbar) pode ser amplificada quando em associação com a incubação em baixa temperatura (-9ºC) e que a combinação destas duas propriedades físicas eleva a solubilização dos agregados em CI, com a consequente elevação dos rendimentos de renaturação de eGFP. Mostramos ainda no presente estudo que a cinética de renaturação de eGFP em 0,69 kbar é proporcional à temperatura de incubação (entre 10ºC e 50ºC). O nível mais elevado de fluorescência foi obtido quando a renaturação de eEGP foi realizada a 20ºC. A taxa de maturação do cromóforo da eGFP é mais fortemente afetada pela temperatura do que a taxa de enovelamento da proteína. Em conclusão, a temperatura de produção dos CI, a temperatura de dissociação dos agregados e a temperatura de enovelamento podem afetar muito o rendimento e a cinética da renaturação de eGFP em alta pressão. Os resultados do presente estudo podem abrir novas perspectivas para melhorias no processo de enovelamento de proteínas a partir de CI utilizando alta pressão. Também neste trabalho descrevemos a renaturação das proteínas de Xac, PilB e os produtos dos genes XAC2810 e XAC3272 nunca antes obtidas na forma solúvel. Os rendimentos de solubilização destas três proteínas foram muito altos, entre 75% e 89%. A proteína PilB renaturada em alta pressão apresentou atividade ATPasica elevada, o que nunca antes foi demonstrado para a PilB de Xac. / In the present work we studied the refolding under high hydrostatic pressure of a mutant form of the green fluorescent protein (eGFP), which only emits the green characteristic fluorescence when in the native folded state. The approach of the present study was focused on controlling the bioactivity of the recombinant protein, the fluorescence, as an alternative for the determination of protein solubility, which is not an ideal indicator of proper protein folding. We studied the action of high pressure in the solubilization of the inclusion bodies (IB) of eGFP produced in bacteria E. coli and in the folding of this protein. The compression of a suspension of eGFP IB at 2.4 kbar for 30 minutes promoted dissociation of aggregates. However, the eGFP folding, monitored by the fluorescence at 509 nm, does not occur in this pressure level. The process of eGFP refolding was evaluated under various decompression conditions after dissociation of the IB at 2.4 kbar. During the gradual decompression, the increase in fluorescence was achieved at pressures ranging between atmospheric pressure and 1.38 kbar. The higher levels of eGFP fluorescence were obtained by incubation for several hours at pressure levels between 0.35 and 0.69 kbar. It is possible that the pressure condition that proved favorable for refolding of eGFP can also be used to favor the folding of other monomeric proteins. Using eGFP as a model, we also found that the IB produced by bacteria grown in a relatively low temperature (37ºC) is more fluorescent, presenting a higher amount of recombinant protein with the characteristic fluorescence at 509 nm, i.e., in its native form, than the IB expressed at higher temperatures (42ºC and 47ºC). The analysis by infrared spectroscopy (FT-IR) also demonstrated that the IB produced at milder temperatures have a higher degree of secondary structure similar to the protein in its native form. Furthermore, the IB produced at 37ºC are also more readily solubilized by the action of high pressure than those produced at the higher temperatures. As expected, the folding of eGFP from IB produced at 37ºC was 25 times more efficient than that obtained using IB produced at 47ºC. In this study we demonstrated that the dissociation of aggregates exerted by the action of high pressure (2.4 kbar) can be amplified by combination with incubation at low temperature (-9ºC) and the association of these two physical properties can be used to increase the solubilization of the aggregates in IB, with a consequent increase in the yield of eGFP refolding. In the present study we also showed that the kinetics of refolding of eGFP is proportional to temperature (10ºC 50ºC). The higher level of fluorescence was obtained when the refolding of eGFP was performed at 20°C. The rate of maturation of the eGFP chromophore is more strongly affected by temperature than the rate of folding of the protein. In conclusion, the temperature of production of IB, the temperature of dissociation of aggregates and the folding temperature can greatly affect the yield and kinetics of refolding of eGFP at high pressure. The results of this study may open new perspectives for improvements in the process of protein folding from IB using high pressure. In this paper we also describe the refolding of the proteins of Xac, PilB and the gene products XAC2810 and XAC3272, which have never before been achieved in soluble form. The yields of solubilization/refolding of these three proteins were very high, between 75% and 89%. The protein PilB refolded at high pressure presented high ATPase activity, which has never been shown for the PilB of Xac.
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Utilização de altas pressões hidrostáticas para o estudo e renaturação de proteínas com estrutura quaternária / Utilization of high hydrostatic pressure for the study and refolding of proteins with quaternary structureRodrigues, Daniella 24 September 2012 (has links)
A produção de proteínas recombinantes é uma ferramenta essencial para a indústria biotecnológica e suporta a expansão da pesquisa biológica moderna. Uma variedade de hospedeiros pode ser utilizada para produzir estas proteínas e dentre eles, as bactérias E. coli são as hospedeiras mais utilizadas. No entanto, a expressão heteróloga de genes em E. coli frequentemente resulta em um processo de enovelamento incompleto que leva ao acúmulo de agregados insolúveis, conhecidos como corpos de inclusão (CI). Altas pressões hidrostáticas são capazes de desfavorecer interações intermoleculares hidrofóbicas e eletrostáticas, levando à dissociação dos agregados e por isso são úteis para solubilizar e renaturar proteínas agregadas em CI. O presente trabalho teve como objetivo o estudo do processo de desagregação dos CI e de renaturação das proteínas oligoméricas subunidade B da toxina colérica (CTB) e região globular da fibra adenoviral (RGFA) utilizando altas pressões hidrostáticas. A toxina colérica (CT) é composta por uma subunidade A e cinco subunidades B combinadas em uma holotoxina AB5. A CTB é a porção pentamérica não tóxica da CT, responsável pela ligação da holotoxina ao receptor gangliosídeo GM1. A fibra do adenovírus é uma proteína homotrimérica que forma parte do capsídeo viral, organizada em três regiões: a cauda N-terminal, a haste central e a região C-terminal (região globular). A RGFA se liga à proteína de membrana CAR nas células hospedeiras e promove a internalização do vírus. Os estudos apresentados neste trabalho demonstraram que a alta pressão hidrostática foi eficaz na desagregação dos CI da CTB e da RGFA. As condições de renaturação foram otimizadas utilizando-se diferentes proporções do par redox glutationa oxidada e reduzida, concentrações de agentes caotrópicos, presença de aditivos e esquemas diferenciados de compressão/descompressão daqueles previamente descritos na literatura. CTB solúvel e pentamérica foi obtida pela compressão da suspensão de CI a 2,4 kbar por 16 horas em tampão TrisHCl 50 mM pH 8,5, 1 mM de tween 20 e descompressão direta seguida de incubação em pressão atmosférica. O rendimento de renaturação da CTB solúvel e pentamérica foi de até 45 % e 288 mg de CTB/litro de cultura bacteriana. Esta proteína apresentou estrutura regular e atividade biológica. RGFA trimérica foi obtida pela compressão da suspensão de CI em tampão TrisHCl 50 mM pH 8,0 e 0,5 M de L-arginina a 2,4 kbar por 1,5 horas e 0,4 kbar por 16 horas antes da completa descompressão. O rendimento de proteína solúvel trimérica da RGFA foi de 4 %, porém não foi possível obter a atividade biológica desta proteína. / The production of recombinant proteins is an essential tool for the biotechnology industry and supports the expansion of modern biological research. Recombinant proteins can be produced by a variety of hosts and among them the bacteria E. coli is the most commonly used. However, the expression of heterologous genes in E. coli often results in an incomplete folding process that leads to the accumulation of insoluble aggregates known as inclusion bodies (IB). The application of high hydrostatic pressure impairs intermolecular hydrophobic and electrostatic interactions of proteins in solution, leading to dissociation of aggregates and is therefore useful tool to solubilize and refold aggregated proteins in IB. This work aimed to study the process of disaggregation of IB and refolding of oligomeric proteins the B subunit of cholera toxin (CTB) and the globular region of the adenoviral fiber (RGFA) using high hydrostatic pressure. The cholera toxin (CT) comprises one A subunit and five B subunits, combined in the AB5 holotoxin. The pentameric CTB is non-toxic moiety of CT which is responsible for binding to the receptor ganglioside GM1 holotoxin. The adenovirus fiber is a homotrimeric protein wich forms part of the viral capsid and it is organized into three regions: the N-terminal tail, the central rod and the C-terminal region (globular region). The RGFA binds to membrane protein CAR in host cells and promotes the internalization of virus. The studies presented here demonstrate that high hydrostatic pressure was effective in the disaggregation of the CTB and RGFA IB. The refolding conditions were optimized using different proportions of the redox couple oxidated and reduced glutathione, concentrations of chaotropic agents, presence of additives and pressure/decompression schemes distinguished from the previously described in the literature. Soluble pentameric CTB was obtained when the suspension of IB were compressed at 2.4 kbar for 16 hours in 50 mM of Tris-HCl buffer pH 8.5, 1 mM of tween 20, followed by direct decompression and incubation at atmospheric pressure. The yield of refolded soluble pentameric CTB was up to 45 % and 288 mg of CTB/ liter of bacterial culture. This protein was shown to presented regular structure and biological activity. Trimeric RGFA was obtained by compression of the suspension of IB in 50 mM of Tris-HCl buffer pH 8.0, 0.5M L-arginine at 2.4 kbar for 1.5 hours and at 0.4 kbar for 16 hours prior to the complete decompression. The yield of soluble trimeric RGFA was 4 %, however this protein did not present biological activity.
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The Mechanotransduction of Hydrostatic Pressure by Mesenchymal Stem CellsSeyedeh Ghazaleh Hosseini (5931062) 17 January 2019 (has links)
<div>Mesenchymal stem cells (MSCs) are responsive to mechanical stimuli that play an essential role in directing their differentiation to the chondrogenic lineage. A better</div><div>understanding of the mechanisms that allow MSCs to respond to mechanical stimuli is important to improving cartilage tissue engineering and regenerative medicine. Hydrostatic pressure (HP) in particular is known to be a primary mechanical force in joints. However, little is known about the underlying mechanisms that facilitate HP</div><div>mechanotransduction. Understanding the signaling pathways in MSCs in transducing HP to a beneficial biologic response and their interrelationship were the focus of this thesis. Studies used porcine marrow-derived MSCs seeded in agarose gel. Calcium ion Ca++ signaling, focal adhesion kinase (FAK) involvement, and sirtuin1 activity were investigated in conjunction with HP application.</div><div><br></div><div><div>Intracellular Ca++ concentration was previously shown to be changed with HP application. In our study a bioreactor was used to apply a single application of HP to the MSC-seeded gel structures and observe Ca++ signaling via live imaging of a fluorescent calcium indicator in cells. However, no fluctuations in Ca++ concentrations were observed with 10 minutes loading of HP. Additionally a problem with the biore actor design was discovered. First the gel was floating around in the bioreactor even without loading. After stabilizing the gel and stopping it from floating, there were still about 16 µm of movement and deformation in the system. The movement and deformation was analyzed for the gel structure and different parts of the bioreactor. </div><div><br></div><div>Furthermore, we investigated the role of FAK in early and late chondrogenesis and also its involvement in HP mechanotransduction. A FAK inhibitor was used on MSCs from day 1 to 21 and showed a dose-dependent suppression of chondrogenesis. However, when low doses of FAK inhibitor added to the MSC culture from day 21 to 42, chondrogenesis was not inhibited. With 4 hour cyclic HP, FAK phosphorylation increased. The beneficial effect of HP was suppressed with overnight addition of the</div></div><div><div>FAK inhibitor to MSC medium, suggesting FAK involvement in HP mechanotransducation by MSCs.</div></div><div><br></div><div>Moreover, sirtuin1 participation in MSC chondrogenesis and mechanotransduction was also explored. The results indicated that overnight sirtuin1 inhibition increased chondrogenic gene expression (Agc, Col2, and Sox9) in MSCs. Additionally, the activity of sirtuin1 was decreased with both 4 hour cyclic hydrostatic pressure and inhibitor application. These two together demonstrated that sirtuin1 inhibition enhances chondrogenesis.</div><div><br></div><div><div>In this research we have investigated the role of Ca++ signaling, FAK involvement, and sirtuin1 activity in the mechanotransduction of HP in MSCs. These understandings about the mechanisms regulating the chondrogenesis with respect to HP could have important implications for cartilage tissue engineering and regenerative studies.</div></div>
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Propriedades vibracionais de nanotubos de carbono de parede tripla. / Vibrational Properties of Triple Walled Carbon NanotubesRafael Silva Alencar 28 February 2012 (has links)
Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O estudo das propriedades eletrÃnicas, mecÃnicas e vibracionais dos nanotubos de carbono com poucas camadas à importante para desenvolver e aprimorar os modelos que descrevem o comportamento dos nanotubos de parede mÃltipla, que à o tipo de nanotubo mais usado em aplicaÃÃes tecnolÃgicas. Diversos estudos teÃricos e experimentais tÃm sido realizados visando o entendimento do comportamento dos nanotubos de carbono de paredes simples e dupla (SWNT e DWNT, respectivamente) submetidos a altas pressÃes hidrostÃticas. Entretanto, o comportamento de nanotubos de carbono de parede tripla (TWNTs) sob tais condiÃÃes à ainda desconhecido. Neste trabalho, apresentamos um estudo de espectroscopia Raman ressonante em TWNTs em funÃÃo da pressÃo hidrostÃtica. Utilizamos Ãleo de parafina (Nujol) e uma mistura de metanol:etanol na proporÃÃo de 4:1 como meios transmissores de pressÃo (PTM) e uma cÃlula de pressÃo de bigorna de diamante (DAC-Diamond Anvil Cell) para a aplicaÃÃo da pressÃo. A evoluÃÃo dos modos radiais (RBM) e tangenciais (Banda G) em funÃÃo da pressÃo foi analisada de forma detalhada e comparada com os resultados existentes para os nanotubos de parede simples e dupla. Os dados de espectroscopia Raman mostram que os efeitos de blindagem internos dos TWNTs sÃo mais acentuados que nos DWNTs. As modificaÃÃes nas intensidades dos modos Raman foram interpretadas como sendo associadas as mudanÃas nas condiÃÃes de ressonÃncia dos modos RBM em funÃÃo da pressÃo. / The study of the electronic, mechanical and vibrational properties of the carbon nanotubes with few layers is important for developing and improving models that could describe the behavior of multi-walled carbon nanotubes (MWNTs), which is the kind of nanotube most used in technological applications. Several theoretical and experimental studies have been conducted for understanding the behavior of single and double walled carbon nanotubes (SWNT and DWNT, respectively) under high hydrostatic pressures. However, the behavior of triple walled carbon nanotubes (TWNTs) under such conditions is still unknown. Here, we present a study of resonance Raman spectroscopy in TWNTs as function of hydrostatic pressure. We used paraffin oil and a mixture of methanol:ethanol in 4:1 ratio as pressure transmitting media (PTM) and a diamond anvil cell (DAC) for applying pressure. The evolution of the radial breathing modes (RBM) and the tangential modes (G band) as a function of pressure was analyzed in detail and compared with existing results for SWNTs and DWNTs. The Raman spectroscopy data show that the shielding effects of inner tubes in TWNTs are more pronounced than in DWNTs. The changes in the intensities of Raman modes were interpreted as being associated with pressure induced changes in the resonance conditions.
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