Aplicações em ressonância magnética na área da neurorradiologia: estudos gerais, estudos vasculares, de suscetibilidade e da dinâmica de fluxo do LCR

Tese de mestrado integrado em Engenharia Biomédica e Biofísica , apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2014

Estabilidade de métodos numéricos LCR para a simulação de escoamento

Pós-graduação em Matematica Aplicada e Computacional - FCT

Teaching and using analogy in law

Analogy plays a central role in legal reasoning, yet how to analogize is poorly taught and poorly practiced. We all recognize when legal analogies are being made: when a law professor suggests a difficult hypothetical in class and a student tentatively guesses at the answer based on the cases she read the night before, when an attorney advises a client to settle because a previous case goes against him, or when a judge adopts one precedent over another on the basis that it better fits the present case. However, when it comes to explaining why certain analogies are compelling, persuasive, or better than the alternative, lawyers usually draw a blank. The purpose of this article is to provide a simple model that can be used to teach and to learn how analogy actually works, and what makes one analogy superior to a competing analogy. The model is drawn from a number of theories of analogy making in cognitive science. Cognitive science is the “long-term enterprise to understand the mind scientifically.” The field studies the mechanisms that are involved in cognitive processes like thinking, memory, learning, and recall; and one of its main foci has been on how people construct analogies. The lessons from cognitive science theories of analogy can be applied to legal analogies to give students and lawyers a better understanding of this fundamental process in legal reasoning.

Proteomic identification of putative microRNA394 target genes in Arabidopsis thaliana identifies MLP family members critical for normal development

Expression of the F-Box protein Leaf Curling Responsiveness (LCR) is regulated by microRNA, miR394, and alterations to this interplay in Arabidopsis thaliana produce defects in leaf polarity and shoot apical meristem (SAM) organisation. Although the miR394-LCR node has been documented in Arabidopsis, the identification of proteins targeted by LCR F-box itself has proven problematic. Here, a proteomic analysis of shoot apices from plants with altered LCR levels identified a member of the Major Latex Protein (MLP) family gene as a potential LCR F-box target. Bioinformatic and molecular analyses also suggested that other MLP family members are likely to be targets for this post-translational regulation. Direct interaction between LCR F-Box and MLP423 was validated. Additional MLP members had reduction in protein accumulation, in varying degrees, mediated by LCR F-Box. Transgenic Arabidopsis lines, in which MLP28 expression was reduced through an artificial miRNA technology, displayed severe developmental defects, including changes in leaf patterning and morphology, shoot apex defects, and eventual premature death. These phenotypic characteristics resemble those of Arabidopsis plants modified to over-express LCR. Taken together, the results demonstrate that MLPs are driven to degradation by LCR, and indicate that MLP gene family is target of miR394-LCR regulatory node, representing potential targets for directly post-translational regulation mediated by LCR F-Box. In addition, MLP28 family member is associated with the LCR regulation that is critical for normal Arabidopsis development.

Adaptive Keeper Design for Dynamic Logic Circuits Using Rate Sensing Technique

The increasing variability in device leakage has made the design of keepers for wide OR structures a challenging task. The conventional feedback keepers (CONV) can no longer improve the performance of wide dynamic gates for the future technologies. In this paper, we propose an adaptive keeper technique called rate sensing keeper (RSK) that enables faster switching and tracks the variation across different process corners. It can switch upto 1.9x faster (for 20 legs) than CONV and can scale upto 32 legs as against 20 legs for CONV in a 130-nm 1.2-V process. The delay tracking is within 8% across the different process corners. We demonstrate the circuit operation of RSK using a 32 x 8 register file implemented in an industrial 130-nm 1.2-V CMOS process. The performance of individual dynamic logic gates are also evaluated on chip for various keeper techniques. We show that the RSK technique gives superior performance compared to the other alternatives such as Conditional Keeper (CKP) and current mirror-based keeper (LCR).

Electromagnetic shielding materials and coatings derived from gelation of multiwall carbon nanotubes in an LCST mixture

Thermally induced demixing in an LCST mixture, polystyrene (PS)/polyvinyl methyl ether] (PVME), was used as a template to design materials with high electrical conductivity. This was facilitated by gelation of multiwall carbon nanotubes (MWNTs) in a given phase of the blends. The MWNTs were mixed in the miscible blends and the thermodynamic driven demixing further resulted in selective localization in the PVME phase of the blends. This was further confirmed by atomic force microscopy (AFM). The time dependent gelation of MWNTs at shallow quench depth, evaluated using isochronal temperature sweep by rheology, was studied by monitoring the melt electrical conductivity of the samples in situ by an LCR meter coupled to a rheometer. By varying the composition in the mixture, several intricate shapes like gaskets and also coatings capable of attenuating the EM radiation in the microwave frequency can be derived. For instance, the PVME rich mixtures can be molded in the form of a gasket, O-ring and other intricate shapes while the PS rich mixtures can be coated onto an insulating polymer to enhance the shielding effectiveness (SE) for EM radiation. The SE of the various materials was analyzed using a vector network analyzer in both the X-band (8.2 to 12 GHz) and the K-u-band (12 to 18 GHz) frequency. The improved SE upon gelation of MWNTs in the demixed blends is well evident by comparing the SE before and after demixing. A reflection loss of -35 dB was observed in the blends with 2 wt% MWNTs. Further, by coating a layer of ca. 0.15 mm of PS/PVME/MWNT, a SE of -15 dB at 18 GHz could be obtained.

Efeitos de raios-X de baixa energia em células mamárias

Doses de radiação de baixa energia podem induzir quebras de dupla fita no DNA assim como também produzir perfis alterados de expressão de genes relacionados a estas lesões. Os danos não reparados ou mal reparados levam a uma maior suscetibilidade à transformação oncogênica já que os efeitos biológicos mais importantes causados pela radiação ionizante são mutação e carcinogênese. As lesões no DNA provocadas pela radiação podem também ocasionar o surgimento micronúcleos e as células podem ser induzidas à apoptose. O objetivo deste trabalho é estudar in vitro a presença de micronúcleos e a apoptose ocasionados por Raios-X de baixa energia. Pretende-se analisar estes efeitos biológicos em relação à energia equivalente à utilizada em exames mamográficos usando duas linhagens estabelecias de células de mama: a MCF-7 (tumoral) e a HB-2 (não-tumoral). As células, em crescimento exponencial, foram irradiadas no equipamento de arranjo experimental de mamografia do LCR/UERJ. A dose de 5Gy na energia de 30 kV foi aplicada com taxa de 0,1 Gy/seg utilizando filtro de 0,03 mm de molibdênio. As irradiações foram realizadas duas vezes, após as irradiações, as células foram incubadas por 4, 24 ou 48 horas e posteriormente coradas com o corante Hoechst33258 para análise em microscopia de fluorescência. Para cada análise, 1000 células foram categorizadas pela morfologia do núcleo. Os resultados mostraram que a HB-2, utilizada neste estudo como célula mamária normal, apresentou maior sensibilidade aos efeitos da radiação, com 37 % das células em apoptose após 4 hs de incubação, enquanto a MCF-7 apresentou 6,5 %. Nas análises após 24 hs foi possível confirmar a radioresistência da MCF-7 tendo sido observadas 11% de células em apoptose no grupo irradiado. Houve um aumento crescente de micronúcleos radioinduzidos nas duas linhagens de acordo com os tempos de incubação. Na análise de 4 hs a HB-2 apresentou 3%, em 24 hs, 8,5% e 48 hs, 11,5 %. Diante destes resultados, foi possível concluir que a energia do feixe de raios-X utilizada na mamografia pode ser capaz de ocasionar aumento de ocorrência de apoptose e geração de micronúcleos nas duas linhagens estudadas

Análise dos Modos de Plasmon em Fibras Ópticas com Contraste Arbitrário de Índices de Refração.

Neste trabalho, é feita a análise dos modos de plasmon que se propagam em um filme metálico que cobre uma fibra óptica generalizada. Os modos de plasmon estudados são: Fuga pela Cobertura (lcv), Ligado Simétrico (Sb), Fuga pelo Núcleo (lcr) e Ligado Assimétrico (ab). Os filmes metálicos, para efeito de comparação, utilizados neste trabalho, são: a prata, o ouro e o paládio. Desenvolveu-se um modelo matemático do fenômeno eletromagnético e um software, que gerou um banco de dados que facilitasse a análise de estruturas, com diversas combinações de parâmetros. Com o banco de dados, foram obtidos diversos gráficos, que permitiram: analisar os modos de plasmon, verificar a atenuação das ondas e o comportamento do campo eletromagnético em cada região da estrutura. As confrontações entre as estruturas com filmes de: prata, ouro e paládio, permitiram concluir que aquelas elaboradas com os filmes de prata e de ouro são as que apresentam menores perdas, portanto, as recomendadas na confecção de sensores. Como a prata é mais acessível que o ouro, aconselha-se a sua utilização. A análise e os resultados deste trabalho são originais na literatura especializada.

控制热力学条件下的矿物、岩石电学性质研究

电导率是重要的地球物理参数。模拟地球内部条件下的矿物、岩石电导率测量是了解地球内部物质组成及物理化学性质的有效手段，同时，还可以为野外大地电磁测量结果的解释提供依据。在YJ-紧装式六面顶压机上，对原有的矿物、岩石电性测量系统进行了进一步的改进；建立了一套以Solartron 1260阻抗／增益一相位分析仪为测试仪器，使用Mo电极和Mo盾来控制样品氧逸度的测量系统，该系统的氧逸度环境为Mo-MoO2，接近Iw缓冲对。石英（人造水晶）、橄榄石、纯橄榄岩、辉石岩、二辉橄榄岩、巨晶辉石的电导率测量是在新的测量系统下获得的，而辉长岩的电导率测量是基于LCR仪为测量仪器的测试系统下获得的。在压力为1-3GPa、温度为675K-160OK、频率为106-0.1Hz、氧逸度为Mo-MoO2的条件下，对不同方向石英（人造水晶）的电学性质进行了研究。复阻抗平面上出现了反映样品本身性质的阻抗弧和反映样品与电极之间扩散的直线。石英的电导率随温度增加而增加，随压力的变化比较微弱。石英的导电机制主要为离子导电，载流子可能为碱金属离子和氢离子，这些碱金属离子和氢离子主要在平行于光轴的通道中运动。在相同的温度和压力条件下，a石英的电导率和c轴的夹角有关，石英的电导率随着夹角的增大而减小，表现出了强烈的各向异性。对“各个方向石英在发生了相变前后的电导率进行了研究，发现Q石英转变为p石英后，电导率并没有突变，仍然随着温度的增加而增加。在压力为1-2GP。、温度为563-1173K、频率为12-105Hz的条件下研究了辉长岩的阻抗。结果发现辉长岩复阻抗的实部随频率的增加而减小，而虚部随频率增加先增大后减小；相角随频率增加而减小。在复阻抗平面上出现了反映颗粒内部的阻抗弧，该阻抗弧出现在高频段。实验室获得辉长岩在地壳的压力和温度（1.0GP。和893K）条件下的电导率值为1.77×-4S／m，而高导层的电导率值为0-01-0-15S／m，二者相差了2-3个数量级，推断辉长岩不能在下地壳形成高导层。在压力为3.0GPa、温度为1299-1600K、频率为106-0.1Hz、氧逸度为Mo-MoO2条件下，对不同颗粒粒度的橄榄石电导率进行了测量。在复阻抗平面上均出现了反映颗粒内部电响应的阻抗弧，这些阻抗弧随着温度的增加而减小。而反映颗粒边界导电机制的阻抗弧并不明显，两种阻抗弧出现在不同的频率范围内，反映颗粒内部导电机制的阻抗弧出现在频率较高的范围内，而反映颗粒边界导电机制的阻抗弧出现在频率相对低的范围内。不同粒度橄榄石在3.0GPa条件下的电导率随着温度的增加而增加，它们的激化烩介于1.03-2.11ev之间。在压力为1-3GPa、温度为1282-1544K、频率为0.1-106Hz、氧逸度为MO-MoO2的条件下，对纯橄榄岩的电导率进行了测量。在复阻抗平面上出现了反映颗粒内部电响应和颗粒边界电响应的阻抗弧。反映颗粒内部导电的阻抗弧出现在较高的频率段，随着温度的增加，这些阻抗弧逐渐收缩。颗粒边界的阻抗弧出现在相对低的频率段。纯橄榄岩的电导率随着温度增加而增加，随压力变化比较微弱。对颗粒边界的电导率研究表明，颗粒边界的电导率高于颗粒内部的电导率，总电导率则小于颗粒内部和颗粒边界的电导率，颗粒边界并没有增强总电导率。纯橄榄岩的激化能为1，62eV，而激化体积为0.67cm3／mol，指前因子为5125加。利用实验所获得的拟合参数，建立了地球内部200-40Okm处的电导率模型，并同地球物理模型进行了对比，在温度和氧逸度的合理波动范围内，实验室电导率模型和地球物理模型吻合的很好。在压力为1-2GPa、温度为1228-1584K、频率为0.1-106Hz、氧逸度为MO-MoO2条件下，测量了天然和热压辉石岩、热压巨晶辉石、二辉橄榄岩的电导率。结果发现，在复阻抗平面上出现了反映颗粒内部电响应和颗粒边界电响应的阻抗弧，反映颗粒内部导电的阻抗弧出现在较高的频率段，随着温度的增加，这些阻抗弧逐渐收缩。颗粒边界的阻抗弧出现在相对低的频率段。辉石岩、二辉橄榄岩、巨晶辉石电导率随着温度增加而增加，随压力变化比较微弱。天然辉石岩和热压辉石岩颗粒边界的电导率高于它们各自颗粒内部的电导率，而总电导率则小于颗粒内部和颗粒边界的电导率，颗粒边界并没有增强总电导率。辉石岩一二辉橄榄岩一纯橄榄岩的电导率依次减小，这可能是与它们的铁含量有关。天然辉石岩的电导率与热压辉石岩的电导率的差异可能与样品中的水（氢）含量的不同有关。

Design and implementation of the embedded capacitance layers for decoupling of wireless sensor nodes

In this paper, the embedded capacitance material (ECM) is fabricated between the power and ground layers of the wireless sensor nodes, forming an integrated capacitance to replace the large amount of decoupling capacitors on the board. The ECM material, whose dielectric constant is 16, has the same size of the wireless sensor nodes of 3cm*3cm, with a thickness of only 14μm. Though the capacitance of a single ECM layer being only around 8nF, there are two reasons the ECM layers can still replace the high frequency decoupling capacitors (100nF in our case) on the board. The first reason is: the parasitic inductance of the ECM layer is much lower than the surface mount capacitors'. A smaller capacitance value of the ECM layer could achieve the same resonant frequency of the surface mount decoupling capacitors. Simulation and measurement fit this assumption well. The second reason is: more than one layer of ECM material are utilized during the design step to get a parallel connection of the several ECM capacitance layers, finally leading to a larger value of the capacitance and smaller value of parasitic. Characterization of the ECM is carried out by the LCR meter. To evaluate the behaviors of the ECM layer, time and frequency domain measurements are performed on the power-bus decoupling of the wireless sensor nodes. Comparison with the measurements of bare PCB board and decoupling capacitors solution are provided to show the improvement of the ECM layer. Measurements show that the implementation of the ECM layer can not only save the space of the surface mount decoupling capacitors, but also provide better power-bus decoupling to the nodes.

Neuronal Survival of the Fittest: The Importance of Aerobic Capacity in Exercise-Induced Neurogenesis and Cognition

It is commonly accepted that aerobic exercise increases hippocampal neurogenesis, learning and memory, as well as stress resiliency. However, human populations are widely variable in their inherent aerobic fitness as well as their capacity to show increased aerobic fitness following a period of regimented exercise. It is unclear whether these inherent or acquired components of aerobic fitness play a role in neurocognition. To isolate the potential role of inherent aerobic fitness, we exploited a rat model of high (HCR) and low (LCR) inherent aerobic capacity for running. At a baseline, HCR rats have two- to three-fold higher aerobic capacity than LCR rats. We found that HCR rats also had two- to three- fold more young neurons in the hippocampus than LCR rats as well as rats from the heterogeneous founder population. We then asked whether this enhanced neurogenesis translates to enhanced hippocampal cognition, as is typically seen in exercise-trained animals. Compared to LCR rats, HCR rats performed with high accuracy on tasks designed to test neurogenesis-dependent pattern separation ability by examining investigatory behavior between very similar objects or locations. To investigate whether an aerobic response to exercise is required for exercise-induced changes in neurogenesis and cognition, we utilized a rat model of high (HRT) and low (LRT) aerobic response to treadmill training. At a baseline, HRT and LRT rats have comparable aerobic capacity as measured by a standard treadmill fit test, yet after a standardized training regimen, HRT but not LRT rats robustly increase their aerobic capacity for running. We found that sedentary LRT and HRT rats had equivalent levels of hippocampal neurogenesis, but only HRT rats had an elevation in the number of young neurons in the hippocampus following training, which was positively correlated with accuracy on pattern separation tasks. Taken together, these data suggest that a significant elevation in aerobic capacity is necessary for exercise-induced hippocampal neurogenesis and hippocampal neurogenesis-dependent learning and memory. To investigate the potential for high aerobic capacity to be neuroprotective, doxorubicin chemotherapy was administered to LCR and HCR rats. While doxorubicin induces a progressive decrease in aerobic capacity as well as neurogenesis, HCR rats remain at higher levels on those measures compared to even saline-treated LCR rats. HCR and LCR rats that received exercise training throughout doxorubicin treatment demonstrated positive effects of exercise on aerobic capacity and neurogenesis, regardless of inherent aerobic capacity. Overall, these findings demonstrate that inherent and acquired components of aerobic fitness play a crucial role not only in the cardiorespiratory system but also the fitness of the brain.

Cambios en la biodisponibilidad de cromo en un suelo inducidos por la incorporación de materia orgánica

p.145-151

Creatine deficiency syndomes : a new model of partial GAMT deficiency affecting brain cell development

Les syndromes de déficiences cérébrales en créatine (CCDS) sont dus à des mutations dans les gènes GATM et G AMT (codant pour les enzymes AGAT et G AMT de la voie de synthèse de créatine) ainsi que SLC6A8 (transporteur de créatine), et génèrent une absence ou une très forte baisse de créatine (Cr) dans le cerveau, mesurée par spectroscopic de résonance magnétique. Les patients CCDS développent des handicaps neurologiques sévères. Les patients AGAT et GAMT peuvent être traités avec des doses importantes de Cr, mais gardent dans la plupart des cas des séquelles neurologiques irréversibles. Aucun traitement efficace n'existe à ce jour pour la déficience en SLC6A8. Bien que de nombreux modèles aient été développés pour comprendre la Cr cérébrale en conditions physiologiques, les pathomécanismes des CCDS ne sont pas encore compris. Des souris transgéniques pour les gènes Gatm, Gamt et Slc6a8 ont été générées, mais elles ne miment que partiellement la pathologie humaine. Parmi les CCDS, la déficience en GAMT est la plus sévère, en raison de l'accumulation cérébrale de l'intermédiaire guanidinoacétate (GAA). Alors que la toxicité cérébrale du GAA a été étudiée par exposition directe au GAA d'animaux adultes sains, les mécanismes de la toxicité du GAA en condition de déficience en GAMT dans le cerveau en développement sont encore inconnus. Le but de ce projet était donc de développer un modèle de déficience en GAMT dans des cultures 3D primaires de cellules nerveuses de rat en agrégats par knock-down du gène GAMT, en utilisant un virus adéno-associé (AAV) induisant le mécanisme d'interférence à l'ARN (RNAi). Le virus scAAV2, à la multiplicité d'infection de 1000, s'est révélé le plus efficace pour transduire tous les types de cellules nerveuses des cultures (neurones, astrocytes, oligodendrocytes), et générer un knock-down maximal de la protéine GAMT de 85% (jour in vitro 18). Cette déficience partielle en GAMT s'est révélée insuffisante pour générer une déficience en Cr, mais a causé l'accumulation attendue de GAA, à des doses comparables aux niveaux observés dans le LCR des patients GAMT. Le GAA a induit une croissance axonale anarchique accompagnée d'une baisse de l'apoptose naturelle, suivis par une induction tardive de mort cellulaire non-apoptotique. Le co-traitement par la Cr a prévenu tous les effets toxiques du GAA. Ce travail montre que l'accumulation de GAA en absence de déficience en Cr est suffisante pour affecter le développement du tissu nerveux, et suggère que des formes de déficiences en GAMT supplémentaires, ne présentant pas de déficiences en Cr, pourraient être découvertes par mesure du GAA, en particulier à travers les programmes récemment proposés de dépistage néonatal de la déficience en GAMT. -- Cerebral creatine deficiency syndromes (CCDS) are caused by mutations in the genes GATM and GAMT (respectively coding for the two enzymes of the creatine synthetic pathway, AGAT and GAMT) as well as SLC6A8 (creatine transporter), and lead to the absence or very strong decrease of creatine (Cr) in the brain when measured by magnetic resonance spectroscopy. Affected patients show severe neurological impairments. While AGAT and GAMT deficient patients can be treated with high dosages of Cr, most remain with irreversible brain sequelae. No treatment has been successful so far for SLC6A8 deficiency. While many models have helped understanding the cerebral Cr pathways in physiological conditions, the pathomechanisms underlying CCDS are yet to be elucidated. Transgenic mice carrying mutations in the Gatm, Gamt and Slc6a8 genes have been developed, but only partially mimic the human pathology. Among CCDS, GAMT deficiency is the most severe, due to the CNS accumulation of the guanidinoacetate (GAA) intermediate. While brain toxicity of GAA has been explored through direct GAA exposure of adult healthy animals, the mechanisms underlying GAA toxicity in GAMT deficiency conditions on the developing CNS are yet unknown. The aim of this project was thus to develop and characterize a GAMT deficiency model in developing brain cells by gene knockdown, by adeno-associated virus (AAV)-driven RNA interference (RNAi) in rat 3D organotypic primary brain cell cultures in aggregates. scAAV2 with a multiplicity of infection of 1000 was shown as the most efficient serotype, was able to transduce all brain cell types (neurons, astrocytes, oligodendrocytes) and to induce a maximal GAMT protein knockdown of 85% (day in vitro 18). Metabolite analysis showed that partial GAMT knockdown was insufficient to induce Cr deficiency but generated the awaited GAA accumulation at concentrations comparable to the levels observed in cerebrospinal fluid of GAMT-deficient patients. Accumulated GAA induced axonal hypersprouting paralleled with inhibition of natural apoptosis, followed by a later induction in non-apoptotic cell death. Cr supplementation led to the prevention of all GAA-induced toxic effects. This work shows that GAA accumulation without Cr deficiency is sufficient to affect CNS development, and suggests that additional partial GAMT deficiencies, which may not show the classical brain Cr deficiency, may be discovered through GAA measurement including by recently proposed neonatal screening programs for GAMT deficiency.