Several research groups have applied elevated hydrostatic pressure to ONH astrocytes cultured on a rigid substrate as an in vitro model for glaucoma. These studies have shown significant biological effects and this hydrostatic pressure model is now becoming generally accepted in the ophthalmic community. However, since the applied pressures were modest the finding of significant biological effects due to pressure alone is surprising. We hypothesized that the application of hydrostatic pressure as described in these studies also altered gas tensions in the culture media. Our goal was to design equipment and carry out experiments to separate the biologic effects of pressure from those of hypoxia on cultured astrocytes. We designed equipment and carried out experiments to subject cultures of DITNC1 astrocytes to the four combinations of two levels of each parameter. We explored the morphology and migration rates of astrocytes, but observed no significant change in any of these properties.

Psidium guajava é uma espécie tropical, frutífera, de grande importância na fruticultura brasileira e é usada em diversas partes do mundo para o tratamento de doenças que acometem a humanidade. Além de sua importância alimentícia e medicinal, P. guajava também se mostrou boa bioindicadora de poluentes atmosféricos como ozônio e SO2 , pois apresenta respostas características e dose-dependentes das concentrações desses poluentes. Mostrou-se, dessa forma, uma espécie interessante para o estudo de respostas de espécies tropicais a elevadas concentrações de CO2 atmosférico. A fim de avaliar se o elevado CO2 seria uma situação estressante para essa espécie, foi realizada uma fumigação de 130 mudas de P. guajava cv. Pedro Sato em quatro câmaras de topo aberto: duas com ar ambiente ((∼390 ppm) e duas com atmosfera enriquecida com CO2 (∼720 ppm). Foram realizadas coletas quinzenais, retirando-se cinco indivíduos de cada câmara, aleatoriamente, que foram separados em raiz, caule e folhas e imediatamente congelados com nitrogênio líquido. Foram analisados componentes relacionados a sinalização e inativação de espécies reativas de oxigênio em folhas: os teores de poliaminas solúveis, ácido ascórbico e glutationa, além de fenóis, flavonoides, proantocianidinas e taninos totais. Além disso, os extratos metanólicos de folhas foram submetidos à CLAE para análises quali e quantitativas dos principais flavonoides. Parâmetros relacionados ao desenvolvimento da planta também foram analisados, como incremento em altura, massa seca, fração de massa, acúmulo de carbono, açúcares solúveis e amido. As análises de poliaminas, ácido ascórbico, glutationa, e compostos fenólicos de folhas não revelaram alterações significativas entre os tratamentos, exceto para a análise de taninos totais onde foi observado um aumento significativo dessas substâncias nas plantas mantidas a elevado CO2 e após 90 dias de fumigação. A análise por CLAE revelou que a maioria dos flavonoides dessa espécie é derivada de quercetina. Com relação aos parâmetros de crescimento, P. guajava não apresentou diferenças significativas de incremento em altura e massa seca, mas apresentaram acúmulo de amido nas folhas. Analises multivariadas foram utilizadas para integrar as 46 variáveis analisadas. Neste estudo viu-se que a situação de elevado CO2 em mudas de P. guajava é favorável, visto que ocorre acúmulo de amido, substâncias anti-herbivóricas (taninos) e, aparentemente, não há alteração no balanço redox. Contudo, é de extrema importância estudos que avaliem alterações concomitantes de parâmetros ambientais, como aumento de temperatura, disponibilidade de água, ozônio e elevado CO2 . A goiabeira apresenta grande importância econômica relacionada à produção e qualidade de seus frutos, dessa forma são necessários estudos que avaliem o efeito do aumento do CO2 na quantidade e qualidade de frutos produzidos por esta espécie. Por ser uma espécie de crescimento lento, períodos de exposição mais longos são aconselháveis para uma melhor analise da susceptibilidade dessa espécie ao elevado CO2 / Psidium guajava is a tropical fruit species of great importance in Brazilian economy; it is also used in several parts of the world as medicinal. Besides its importance as medicinal and nutritional species, P. guajava has also showed to be a good bioindicator of air pollutants, such as ozone and SO2 . It presents characteristic responses, dose-dependent of the concentrations of these pollutants. Therefore, this species seems to be interesting to study the responses of a tropical species to high concentrations of atmospheric CO2 . In order to assess whether the high CO2 would be a stressful situation for this species, we performed a fumigation experiment using 130 seedlings of P. guajava cv. Pedro Sato in four open-top chambers: two with ambient air (∼ 390 ppm CO2 ) and two with elevated CO2 (∼ 720 ppm). Biweekly, five individuals of each chamber were randomly collected, separated into root, stem and leaves and immediately frozen with liquid nitrogen. Were analyzed parameters related to signaling and inactivation of reactive oxygen species in leaves: levels of soluble polyamines, ascorbate and glutathione, as well as contents of phenols, flavonoids, tannins and proanthocyanidins. Furthermore, the methanol extracts were subjected to HPLC for quantitative and qualitative analysis of the major flavonoids. Parameters related to the development of the plant were also analyzed, as height increment, dry mass, mass fraction, carbon accumulation, soluble sugars and starch accumulation. The analysis of polyamines, ascorbate, glutathione and phenolic compounds of leaves, showed no significant changes when comparing treatments, except for the analysis of tannins which was observed a significant increase of these substances in plants at high CO2 after 90 days of fumigation. HPLC analysis revealed that most of flavonoid composition is quercetin derivatives. The growth parameters showed no significant differences in height increment and dry mass, but the leaves showed accumulation of starch after CO2 fumigation. Multivariate analyzes were used to integrate the 46 variables analyzed. In this study it was seen that the elevated CO2 seem to be favorable to seedlings of P. guajava, once there is an accumulation of starch, antiherbivore substances (tannins) and, apparently, there was no change in the redox balance. However, it is extremely important to perform studies that evaluate concomitant changes in environmental conditions, such as increased temperature, availability of water, ozone and elevated CO2 . Guava has great economic importance in Brazil, fact related to the production and quality of its fruits, so more studies are needed to assess the effect of increased CO2 in the quantity and quality of fruits produced by this species. As a slow-growing species, longer exposure periods are recommended to better analyze the susceptibility of this species to elevated CO2

The concentration of atmospheric carbon dioxide (CO2) in the atmosphere has increased by 35% since pre-industrial levels. Projections for the next 100 years indicate an increase to levels between 490 and 1260 parts per million by volume (ppm) of CO2, equating to a 75 % to 350 % increase in concentration since the year 1750. Associated with this increase in [CO2] will be a 1.4 to 5.8?? C increase in lower atmospheric temperature. While past research has attempted to address the effects of such climatic changes on individual plant responses, predictions of plant responses at the ecosystem level are still highly uncertain. Difficulties lie in the enormous variation of plant responses to climate change variables among and within species, and between and within environmental conditions. Past research assumed that plants using either the C3 or C4 metabolic pathways would respond differently but predictably to climate-change variables based on their metabolic pathway. Recent evidence has suggested however, that the added interactions of external environmental variables and species-specific sensitivities to climate change make it difficult to predict plant and ecosystem responses to climate change. To investigate the mechanisms behind responses of Australian grasses to climate change, 2 pot experiments was conducted using growth cabinets to compare the effect of elevated CO2 and water-limitation on the invasive C4 grassland plant, Eragrostis curvula (E. curvula), native Australian C3 grassland plant, Austrodanthonia racemosa (A. racemosa), and wheat species, Triticum aestivum (T. aestivum). The experiment was run at ambient levels of CO2 maintained at 390 ppm compared to elevated levels of 740 ppm. Imposed restrictions to water supply consisted of gradually drying the soil down to 30 % available soil water (ASW) followed by re-wetting to 50 % ASW. Well-watered conditions for the experiment consisted of gradually drying the soil down to 50 % ASW, followed by rewetting to 95 % ASW. Plants were grown in mixtures and monocultures, consisting of 9 plants equally spaced in a grid design. The three significant findings of the thesis were that: 1) the metabolic pathway (C3 versus C4) was not always an accurate predictor of biomass accumulation under elevated CO2 in the plants studied. Previous research suggested that CO2-stimulation of photosynthesis in C3 plants would lead to greater increases in biomass under elevated CO2 compared to C4 plants, though both C3 and C4 plants could benefit from any reduction in stomatal conductance under dry conditions at elevated CO2. The results from the experiments in this thesis showed a strongly significant biomass response to elevated CO2 in both dry and wet conditions for C4 grass E. curvula. The C3 grass A. racemosa in dry conditions, did not. It was speculated that without the CO2-induced water conservation effect, the C3 grass experienced photosynthetic down-regulation and this precluded a positive biomass response under elevated CO2. 2) the magnitude and direction of biomass response to elevated CO2 was dependant on factors such as resource-availability and the phenotypic variability of the plants species. 3) critical analysis of results from this thesis, combined with past research on plant responses under elevated CO2 showed a tendency for researchers to repeatedly test plants from the Poaceae family, or close relatives of the Poaceae family. As a result, when past data were corrected for this lack of independence, there was no relationship between the evolution of the C3 and C4 metabolic pathway and biomass response to elevated CO2. Instead, other factors (such as growth rate, plant height, leaf number, etc) were presented as being more important in determining biomass response. These observations were supported by results found in this thesis.

Anxiety disorders are one of the most prevalent mental disorders in the world. While normal anxiety serves as an important protective mechanism, pathological anxiety characteristic of an anxiety disorder is both maladaptive and disruptive. The majority of studies have focused on the neurotransmitter systems associated with the actions of known anxiety drugs. This focus may likely limit the exploration of mechanisms underlying anxiety disorders. This project aims to examine changes in gene expression that may underlie higher or lower levels of inherent anxiety. Using a well-established behavior test for anxiety, the elevated plus maze, we identified male Wistar rats exhibiting inherently high- or low-anxiety levels. Brain regions known to mediate anxiety, the amygdala, hippocampus and nucleus accumbens, were dissected and total mRNA isolated. The mRNA was converted to cDNA via reverse transcription-polymerase chain reaction (RT-PCR). Then, the cDNA was used in suppression subtractive hybridization, a technique used to compare two complete populations of cDNAs and identify cDNAs that are upregulated in one population in relation to the other. In this project suppression subtractive hybridization was used to compare high- and low-anxiety cDNA populations. The upregulated cDNAs were amplified in a PCR reaction that enables rare transcripts to be identified. The PCR products from the suppression subtractive hybridization were cloned and used to create two cDNA libraries for high- and low-anxiety related genes. These clones were sequenced to show over 1000 genes upregulated in high- and low-anxiety. The gene list was then subjected to bioinformatic analysis to identify one candidate to be studied in further detail. <p>The prion protein was identified as a potential candidate. Examination of the literature sparked an interest in studying other prion-like proteins, more specifically the cytoplasmic polyadenylation element binding protein (CPEB). The CPEB protein is a potent regulator of mRNA translation in both mature oocytes and the adult brain. While unphosphorylated the CPEB protein keeps specific mRNAs dormant in the cytoplasm. In its phosphorylated form CPEB catalyzes polyadenylation of the mRNA, leading to protein synthesis. p*PCR was used to show the presence of CPEB mRNA transcripts in the rat hippocampus. CPEB protein expression was examined in the brain samples isolated from control, high- and low-anxiety rats. It was found that CPEB was significantly upregulated in high- and low-anxiety rats compared to control. The protein expression of an upstream kinase, Aurora A kinase, and a downstream target, Calcium/Calmodulin Dependent Kinase II (CaMKII), was also investigated. The results from Aurora A kinase were inconclusive. CaMKII, on the other hand, was significantly upregulated in high-anxiety over both control and low-anxiety. These results suggest that CPEB may catalyze increased translation of mRNAs in high-anxiety while acting as a repressor of those same mRNAs in low-anxiety. <p>Recent studies have suggested that CPEB protein plays an important role in synaptic plasticity. The regulation of synaptic plasticity, and its impact on learning and memory, is believed to be a key mechanism behind the maintenance of anxiety disorders. Therefore the results of this study suggest a new molecular mechanism in the development of anxiety disorders.

Anxiety disorders are one of the most prevalent mental disorders in the world. While normal anxiety serves as an important protective mechanism, pathological anxiety characteristic of an anxiety disorder is both maladaptive and disruptive. The majority of studies have focused on the neurotransmitter systems associated with the actions of known anxiety drugs. This focus may likely limit the exploration of mechanisms underlying anxiety disorders. This project aims to examine changes in gene expression that may underlie higher or lower levels of inherent anxiety. Using a well-established behavior test for anxiety, the elevated plus maze, we identified male Wistar rats exhibiting inherently high- or low-anxiety levels. Brain regions known to mediate anxiety, the amygdala, hippocampus and nucleus accumbens, were dissected and total mRNA isolated. The mRNA was converted to cDNA via reverse transcription-polymerase chain reaction (RT-PCR). Then, the cDNA was used in suppression subtractive hybridization, a technique used to compare two complete populations of cDNAs and identify cDNAs that are upregulated in one population in relation to the other. In this project suppression subtractive hybridization was used to compare high- and low-anxiety cDNA populations. The upregulated cDNAs were amplified in a PCR reaction that enables rare transcripts to be identified. The PCR products from the suppression subtractive hybridization were cloned and used to create two cDNA libraries for high- and low-anxiety related genes. These clones were sequenced to show over 1000 genes upregulated in high- and low-anxiety. The gene list was then subjected to bioinformatic analysis to identify one candidate to be studied in further detail. <p>The prion protein was identified as a potential candidate. Examination of the literature sparked an interest in studying other prion-like proteins, more specifically the cytoplasmic polyadenylation element binding protein (CPEB). The CPEB protein is a potent regulator of mRNA translation in both mature oocytes and the adult brain. While unphosphorylated the CPEB protein keeps specific mRNAs dormant in the cytoplasm. In its phosphorylated form CPEB catalyzes polyadenylation of the mRNA, leading to protein synthesis. p*PCR was used to show the presence of CPEB mRNA transcripts in the rat hippocampus. CPEB protein expression was examined in the brain samples isolated from control, high- and low-anxiety rats. It was found that CPEB was significantly upregulated in high- and low-anxiety rats compared to control. The protein expression of an upstream kinase, Aurora A kinase, and a downstream target, Calcium/Calmodulin Dependent Kinase II (CaMKII), was also investigated. The results from Aurora A kinase were inconclusive. CaMKII, on the other hand, was significantly upregulated in high-anxiety over both control and low-anxiety. These results suggest that CPEB may catalyze increased translation of mRNAs in high-anxiety while acting as a repressor of those same mRNAs in low-anxiety. <p>Recent studies have suggested that CPEB protein plays an important role in synaptic plasticity. The regulation of synaptic plasticity, and its impact on learning and memory, is believed to be a key mechanism behind the maintenance of anxiety disorders. Therefore the results of this study suggest a new molecular mechanism in the development of anxiety disorders.

The purpose of thesis aims to investigate the mechanical behavior and properties of a centrally notched hybrid Al alloy/Carbon-Fiber/PEEK(APC-2) laminate at elevated temperature. The high performance hybrid composite laminates of 0.5mm Aluminum alloy sheets sandwiched by APC-2 cross-ply and guasi-isotropic laminates were fabricated. The prepregs of APC-2 were stacked into cross-ply [0/90]s and quasi-isotropic [0/45/90/-45] laminates spread uniformly with nanoparticles SiO2. The sheet surface was treated by chromic acid anodic method to achieve perfectly bonding with matrix PEEK. The modified diaphragm curing process was adopted to fabricate Al/APC-2 hybrid nanocomposite laminates. The panels were cut into the specimens and then drilled an diameter hole in the center with diameters of 1,2,4,6 mm. The MTS 810 material testing machine was used to conduct the tension and fatigue tests. In addition, the MTS 651 environmental chamber was installed to control and keep the specific testing temperatures, such as ,25¢XC(RT), 75¢XC, 100¢XC, 125¢XC and 150¢XC. At first, the nominal stress(£mnom) and stress-strain diagram were obtained due to static tension tests at elevated temperature. The constant stress amplitude tension-tension cyclic tests were carried out by using load-control mode at a sinusoidal loading with frequency of 5Hz and stress ratio R=0.1. The received fatigue data were plotted in normalized S-N curves at variously elevated temperature. For the tensile tests, at the same temperature the nominal stress of cross-ply specimens was higher than that of quasi-isotropic specimens. Comparing with the notched and unnotched of cross-ply specimens, the nominal stress of notched specimens was about 60% to 80% that of unnotched specimens. Besides, as for the notched and unnotched quasi-isotropic specimens, the nominal stress of notched specimens was about 75% to 85% that of unnotched specimens. Then, the fatigue life and stress-cycles (S-N) curves of notched specimens were obtained often tension-tension fatigue tests. In the case of the same loading, notched specimens possess worse fatigue behavior, but in the same normalized stress ratio, the S-N curves of the unnotched were below the notched ones. The fatigue resistance of notched samples decrease as the temperature rising.

The aims of this thesis are fabrication of Ti/APC-2 hybrid nanocomposite laminates and investigation of their mechanical properties at elevated temperature. The prepregs of APC-2 were stacked into cross-ply [0/90]s and quasi-isotropic [0/45/90/-45] laminates spread uniformly with nanoparticles SiO2. The sheet surface was treated by chromic acid anodic method to achieve perfectly bonding with matrix PEEK. The prepregs were sandwiched with the Ti alloy sheets. The modified diaphragm curing process was adopted to produce Ti/APC-2 hybrid nanocomposite laminates. The nanocomposite laminates were a five-layer composite with two 0.55 mm thick APC-2 layers sandwiched by three 0.5 mm thick Gr.1 titanium alloy sheets. The MTS 810 material testing machine was used to conduct the tension and fatigue tests. In addition, the MTS 651 environmental chamber was installed to control and keep the experimental temperature, such as 25¢XC, 75¢XC, 100¢XC, 125¢XC and 150¢XC. The mechanical proper&not;ties, such as ultimate tensile strength, longitudinal stiffness of cross-ply and quasi-isotropic nanocomposite laminates, were obtained from the static tensile test. The stress-strain diagrams were plotted in the corresponding temperature. The constant stress amplitude tension-tension cyclic tests were carried out by using load-control mode at a sinusoidal loading with frequency of 5Hz and stress ratio R=0.1. The received fatigue data were plotted in normalized S-N curves at variously elevated temperature. From the summarized results, some conclusions were made. First, the ultimate strength of Ti/APC-2 nanocomposits was better than Ti/APC-2 composites at room temperature; Second, Both two type nanocomposite laminates¡¦ ultimate strength and S-N curves go downwards as temperature rising, especially at 150¢XC; Third, The fatigue tensile strength of both hybrid composite laminates was the lowest at 150¢XC. Fourth, Ti/APC-2 quasi-isotropic nanocomposite laminates had better fatigue resistance than Ti/APC-2 cross-ply nanocomposite laminates. Finally,The longitudinal stiffness was in good agreement with prediction by using the modified ROM because of the changed curve fitting ranges.

The innovative Al/APC-2 hybrid nanocomposite fiber metal laminates (FMLs) were successfully fabricated. To overcome the usual problem of delamination, the Al alloy 2024-T3 thin sheets were treated by chromic acid anodic (CAA) method to achieve perfectly bonding with matrix PEEK eventually. It was found much better than the previously surface treatment method of CrO3-based chemical etching. A systematic study of hybrid specimens subjected to both static tensile and fatigue tests was conducted at elevated temperatures to obtain their mechanical properties, fatigue lives and failure mechanisms. From the tensile tests, the mechanical properties of Al/APC-2 hybrid cross-ply and quasi-isotropic nanocomposite FLMs at elevated temperatures were received, such as ultimate tensile strength and longitudinal stiffness. Also, the predicted stress-strain curves was proposed and in good agreement with experimental data. The average values of received notched strength were affected significantly by stress concentration and high temperature. The modified point stress criterion (PSC) was used with the varied characteristic length dependent on nature of material and specimen geometry. The predicted notched strengths by the modified PSC model were not only precisely validated, but extended to the application at elevated temperatures. The received fatigue data were plotted in S-N curves at variously elevated temperatures. The predictions of fatigue life curves were also presented and verified. The predicted S-N curves were compared with experimental data and found quite accurate.

The work aims to manufacture AS-4/PEEK APC-2 nano-composite laminates first. We used the prepreg form of AS-4 Graphite/PEEK laminae to make APC-2 laminates of 2 mm thick with two lay-ups of cross-ply and quasi-isotropic totally 16 plies by a hot press via the modified diaphragm curing. The nano-particles SiO2 with the average diameter of 15¡Ó5 nm were uniformly spread in the specific interfaces of laminate. From mechanical testing it is found that the nanocomposite specimens of spreading 10 plies nanoparticles (3% by wt. of matrix) possesses the highest mechanical properties. we see that in cross-ply specimens the ultimate strength increases 10.91 % and stiffness 6.7 %; while in quasi-isotropic specimens the ultimate strength increases 12.48 % and stiffness 19.93 %. Second, repeat the tensile tests at 50, 75, 100, 125, 150¢J to receive respective stress-strain curve , strength and stiffness. At elevated temperatures the ultimate strength decreases slightly below 75¢J and the elastic modulus reduces slightly below 125¢J, however, both properties degrade highly at 150¢J ( Tg) for two layups generally. Finally, the constant stress amplitude tension-tension cyclic testing was conducted. It is found that both the stress-cycles (S-N) curves are very close below 104 cycles for cross-ply laminates w/wo nanoparticles, and the S-N curve of nano-laminate slightly bent down after 105 cycles. Whilst in quasi-isotropic laminates, the S-N curve of nano-laminate is always slightly below that of APC-2 laminate through the life.

ABSTRACT PEEK matrix reinforced by carbon fibers as one thermoplastic composite material is studied. Thermoplastic composites have the advantages of high specific stiffness and strength, longer fatigue life, good resistance to moisture absorption and high temperature condition. The thesis is aimed to investigate the mechanical properties and fracture mechanism of the centrally notched AS-4/PEEK composite laminates subjected to tension-tension fatigue loading at elevated temperature. We use three common types of laminates, such as cross-ply , quasi-isotropic and angle-ply . After centrally notched, we first obtain the base-line data of mechanical properties by tensile tests at five different temperatures, such as 25¢J¡B75¢J¡B100¢J¡B125¢J¡B150¢J. Then, the fatigue tests are conducted, we receive the fatigue strength and life and establish the stress-life curves. The fatigue characteristics and fracture mechanism of a centrally notched composite laminate at elevated temperature are also recorded and observed. The empirical results can be concluded as follows. At the same temperature, the laminate of cross-ply possesses the largest ultimate strength and fatigue strength, quasi-isotropic the second angle-ply the smallest. As for the elastic modulus, the laminate of cross-ply is larger than that of quasi-isotropic. However, the large strain of angle-ply is within the plastic range that is out of the limit of the study. Thus, a further investigation is needed for angle-ply laminates alone. After centrally notched, the net area is reduced of the specimen, and then the elastic modulus is raised and the ultimate strength and fatigue strength of composite materials are lower. As the temperature increasing, the ultimate strength, fatigue strength and elastic modulus are all decreasing.