Neurotoxic activity and ultrastructural changes in muscles caused by the brown widow spider Latrodectus geometricus venom

Brown widow spider (Latrodectus geometricus) venom (BrWSV) produces few local lesions and intense systemic reactions such as cramps, harsh muscle pains, nausea, vomiting and hypertension. Approximately 16 protein bands under reducing conditions and ~ 14 bands under non-reducing conditions on a 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis were observed. Neurotoxic clinical manifestations were confirmed in vivo, while proteolytic activity was demonstrated on gelatine film. Severe ultrastructural damages in mice skeletal muscles were observed at 3, 6, 12 and 24 h postinjection with at total of 45 µg of venom protein. Infiltration of eosinophils and ruptures of the cellular membranes were observed in the muscles along with swelling of the nuclear cover and interruption of the collagen periodicity. Altered mitochondrias and autophage vacuoles, nuclear indentation and mitochondria without cristae, slight increment of intermyofibrillar and subsarcolemic spaces and myelinic figures formation were also observed. In the capillary, endothelial membrane unfolding into the lumen was noticed; along with myelinic figures compatible with a toxic myopathy. Swollen sarcotubular systems with lysis of membrane, intense mitochondria autophagia and areas without pinocytic vesicles were observed. Swollen mitochondria surrounded by necrotic areas, myofibrillar disorganization and big vacuolas of the sarcotubular system, degenerated mitochondrium with formation of myelinic figure was seen. Glycogenosomes with small particulate, muscle type glycogen was noticed. Autophagic vacuole (autophagolysosomes) and necrotic areas were also noticed. These damages may be due to interactive effects of the multifactorial action of venom components. However, Latrodectus geometricus venom molecules may also be utilized as neuro therapeutic tools, as they affect neuronal activities with high affinity and selectivity. To our knowledge, the present study is the first ultrastructural report in the literature of muscle injuries and neurological and proteolytic activities caused by BrWSV.

Aquisição por Visão Artificial e Medição Automática de Micro Dureza Vickers

O ensaio de dureza, e mais concretamente o ensaio de micro dureza Vickers, é no universo dos ensaios mecânicos um dos mais utilizados quer seja na indústria, no ensino ou na investigação e desenvolvimento de produto no âmbito das ciências dos materiais. Na grande maioria dos casos, a utilização deste ensaio tem como principal aplicação a caracterização ou controlo da qualidade de fabrico de materiais metálicos. Sendo um ensaio de relativa simplicidade de execução, rapidez e com resultados comparáveis e relacionáveis a outras grandezas físicas das propriedades dos materiais. Contudo, e tratando-se de um método de ensaio cuja intervenção humana é importante, na medição da indentação gerada por penetração mecânica através de um sistema ótico, não deixa de exibir algumas debilidades que daí advêm, como sendo o treino dos técnicos e respetivas acuidades visuais, fenómenos de fadiga visual que afetam os resultados ao longo de um turno de trabalho; ora estes fenómenos afetam a repetibilidade e reprodutibilidade dos resultados obtidos no ensaio. O CINFU possui um micro durómetro Vickers, cuja realização dos ensaios depende de um técnico treinado para a execução do mesmo, apresentando todas as debilidades já mencionadas e que o tornou elegível para o estudo e aplicação de uma solução alternativa. Assim, esta dissertação apresenta o desenvolvimento de uma solução alternativa ao método ótico convencional na medição de micro dureza Vickers. Utilizando programação em LabVIEW da National Instruments, juntamente com as ferramentas de visão computacional (NI Vision), o programa começa por solicitar ao técnico a seleção da câmara para aquisição da imagem digital acoplada ao micro durómetro, seleção do método de ensaio (Força de ensaio); posteriormente o programa efetua o tratamento da imagem (aplicação de filtros para eliminação do ruído de fundo da imagem original), segue-se, por indicação do operador, a zona de interesse (ROI) e por sua vez são identificadas automaticamente os vértices da calote e respetivas distâncias das diagonais geradas concluindo, após aceitação das mesmas, com o respetivo cálculo de micro dureza resultante. Para validação dos resultados foram utilizados blocos-padrão de dureza certificada (CRM), cujos resultados foram satisfatórios, tendo-se obtido um elevado nível de exatidão nas medições efetuadas. Por fim, desenvolveu-se uma folha de cálculo em Excel com a determinação da incerteza associada às medições de micro dureza Vickers. Foram então comparados os resultados nas duas metodologias possíveis, pelo método ótico convencional e pela utilização das ferramentas de visão computacional, tendo-se obtido bons resultados com a solução proposta.

Kinetic, Structural, and EPR Studies Reveal That Aldehyde Oxidoreductase from Desulfovibrio gigas Does Not Need a Sulfido Ligand for Catalysis and Give Evidence for a Direct Mo-C Interaction in a Biological System

J. Am. Chem. Soc., 2009, 131 (23), pp 7990–7998 DOI: 10.1021/ja809448r

Correlating EPR and X-ray structural analysis of arsenite-inhibited forms of aldehyde oxidoreductase

J Biol Inorg Chem (2007) 12:353–366 DOI 10.1007/s00775-006-0191-9

Functional and structural studies of two enzymes: membrane bound kinase and Z-DNA/Z-RNA-binding protein

Dissertação para obtenção do Grau de Doutor em Sistemas de Bioengenharia

Italian interlocking directorates: structure, evolution and cross-country comparisons

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Who owns the largest firms around the world?

In this research paper we evaluate how corporate control around the world is defined and which variables, related to a firm's characteristics and the countries' infrastructures, influences this. We find that there is a small number of countries where firms are widely held. The role of financial institutions seems to be different in civil and common law-based countries. While they seem to act as a monitor of management in common law-based countries, in civil law countries they act as a monitor of large shareholders. Finally, we find that firm's size is the most important determinant of ultimate owners.

Late-medieval construction site management at the Monastery of Jerónimos in Lisbon

Through the analysis of the exceptional accounting documents of 1517 related to the construction of the Monastery of Jerónimos (Lisbon), this paper discusses the main characteristics of a new model of construction site organization. In the later Middle Ages we can find, among others, two main models of constructing site organization. One, older and more widespread, consisted in a centralized and pyramidal management model. The other, apparently more recent, was based in the existence of several autonomous teams working simultaneously, each one responsible for building a specific part or section of the building. This paper describes and discusses this new organizational model as it was adopted and implemented by João de Castilho (1470–1552) for the construction of the Monastery of Jerónimos in 1517, probably for the first time in Portugal, but with some parallels in other places in Europe.

Structural, mechanical and piezoelectric properties of polycrystalline AlN films sputtered on titanium bottom electrodes

Polycrystalline AlN coatings deposited on Ti-electrodes films were sputtered by using nitrogen both as reactive gas and sputtering gas, in order to obtain high purity coatings with appropriate properties to be further integrated into wear resistance coatings as a piezoelectric monitoring wear sensor. The chemical composition, the structure and the morphology of the films were investigated by X-ray diffraction, X-ray photoelectron spectroscopy, scanning electron microscopy and atomic force microscopy techniques. These measurements show the formation of highly (101), (102) and (103) oriented AlN films with good piezoelectric and mechanical properties suitable for applications in electronic devices. Through the use of lower nitrogen flow a densification of the AlN coating occurs in the microstructure, with an improvement of the crystallinity along with the increase of the hardness. Thermal stability of aluminum nitride coatings at high temperature was also examined. It was found an improvement of the piezoelectric properties of the highly (10x) oriented AlN films which became c-axis (002) oriented after annealing. The mechanical behavior after heat treatment shows an important enhancement of the surface hardness and Young’s modulus, which decrease rapidly with the increase of the indentation depth until approach constant values close to the substrate properties after annealing. Thus, thermal annealing energy promotes not only the rearrangement of Al–N network, but also the occurrence of a nitriding process of unsaturated Al atoms which cause a surface hardening of the film.

“PhenoWorld”: a new multidimensional strategy for studying behaviour in rodents

Tese de Doutoramento em Ciências da Saúde

Stress induced risk-aversion is reverted by D2/D3 agonist in the rat

Stress exposure triggers cognitive and behavioral impairments that influence decision-making processes. Decisions under a context of uncertainty require complex reward-prediction processes that are known to be mediated by the mesocorticolimbic dopamine (DA) system in brain areas sensitive to the deleterious effects of chronic stress, in particular the orbitofrontal cortex (OFC). Using a decision-making task, we show that chronic stress biases risk-based decision-making to safer behaviors. This decision-making pattern is associated with an increased activation of the lateral part of the OFC and with morphological changes in pyramidal neurons specifically recruited by this task. Additionally, stress exposure induces a hypodopaminergic status accompanied by increased mRNA levels of the dopamine receptor type 2 (Drd2) in the OFC; importantly, treatment with a D2/D3 agonist quinpirole reverts the shift to safer behaviors induced by stress on risky decision-making. These results suggest that the brain mechanisms related to risk-based decision-making are altered after chronic stress, but can be modulated by manipulation of dopaminergic transmission.

Pure elastic contact force models

"Series: Solid mechanics and its applications, vol. 226"

Acoustic emission analysis of the effect of a 2D wedge shaped blade on the compact bone cutting process

Surgeons may use a number of cutting instruments such as osteotomes and chisels to cut bone during an operative procedure. The initial loading of cortical bone during the cutting process results in the formation of microcracks in the vicinity of the cutting zone with main crack propagation to failure occuring with continued loading. When a material cracks, energy is emitted in the form of Acoustic Emission (AE) signals that spread in all directions, therefore, AE transducers can be used to monitor the occurrence and development of microcracking and crack propagation in cortical bone. In this research, number of AE signals (hits) and related parameters including amplitude, duration and absolute energy (abs-energy) were recorded during the indentation cutting process by a wedge blade on cortical bone specimens. The cutting force was also measured to correlate between load-displacement curves and the output from the AE sensor. The results from experiments show AE signals increase substantially during the loading just prior to fracture between 90% and 100% of maximum fracture load. Furthermore, an amplitude threshold value of 64dB (with approximate abs-energy of 1500 aJ) was established to saparate AE signals associated with microcracking (41 – 64dB) from fracture related signals (65 – 98dB). The results also demonstrated that the complete fracture event which had the highest duration value can be distinguished from other growing macrocracks which did not lead to catastrophic fracture. It was observed that the main crack initiation may be detected by capturing a high amplitude signal at a mean load value of 87% of maximum load and unsteady crack propagation may occur just prior to final fracture event at a mean load value of 96% of maximum load. The author concludes that the AE method is useful in understanding the crack initiation and fracture during the indentation cutting process.

Mechanische Charakterisierung von Konstruktionskeramik im oberflächennahen Bereich

Engineering ceramics, indentation methods, crack-resistance, Rayleigh waves, acoustic microscopy, machining parameters, green ceramic bodies, ultrasonic contact impedance, acoustic impedance, residual stress

Comportamiento tribomecánico de sistemas de sustrato: recubrimiento duros

En aplicaciones como la conformación en frío, donde los metales duros recubiertos con películas de naturaleza cerámica son ampliamente empleados, la existencia de un contacto mecánico repetitivo induce tensiones Hertzianas y origina el fallo por fatiga. En este trabajo, se investigan diversos recubrimientos cerámicos depositados por deposición física desde fase vapor sobre calidades diferentes de metal duro y un acero rápido pulvimetalúrgico para evaluar sus respectivas respuesta al contacto y comportamiento a fatiga. El trabajo experimental incluye la caracterización de los sistemas mediante ensayos de rayado y nanoindentación y la evaluación de las curvas tensión-deformación de indentación esférica de los sustratos, tanto desnudos como recubiertos, poniendo especial atención en determinar las tensiones de contacto críticas asociadas a la deformación plástica y a la aparición de grietas circulares en la superficie recubierta. A este estudio, le siguen numerosos ensayos a fatiga a cargas inferiores a aquéllas identificadas como críticas bajo carga monotónica y para un número de ciclos comprendido entre 1.000 y 1.000.000 de ciclos. Los resultados experimentales indican que las películas cerámicas no parecen desempeñar un papel relevante en la aparición de la cedencia plástica, siendo la deformación plástica global controlada por la deformación del sustrato. No obstante, para tensiones elevadas de indentación durante el régimen plástico, existe la aparición de grietas circulares en los recubrimientos cerámicos. Además, la aparición de las mismas es sensible a la fatiga por contacto. Este análisis mecánico se complementa con una inspección detallada del daño generado en profundidad y superficie.