Trajectory Mapping ("TM''): A New Non-Metric Scaling Technique

Trajectory Mapping "TM'' is a new scaling technique designed to recover the parameterizations, axes, and paths used to traverse a feature space. Unlike Multidimensional Scaling (MDS), there is no assumption that the space is homogenous or metric. Although some metric ordering information is obtained with TM, the main output is the feature parameterizations that partition the given domain of object samples into different categories. Following an introductory example, the technique is further illustrated using first a set of colors and then a collection of textures taken from Brodatz (1966).

Keeping Secrets in Hardware: the Microsoft Xbox(TM) Case Study

This paper discusses the hardware foundations of the cryptosystem employed by the Xbox(TM) video game console from Microsoft. A secret boot block overlay is buried within a system ASIC. This secret boot block decrypts and verifies portions of an external FLASH-type ROM. The presence of the secret boot block is camouflaged by a decoy boot block in the external ROM. The code contained within the secret boot block is transferred to the CPU in the clear over a set of high-speed busses where it can be extracted using simple custom hardware. The paper concludes with recommendations for improving the Xbox security system. One lesson of this study is that the use of a high-performance bus alone is not a sufficient security measure, given the advent of inexpensive, fast rapid prototyping services and high-performance FPGAs.

Mapeamento e descrição do padráo de uso e cobertura da terra em municípios do sudoeste goiano a partir de imagens orbitais TM/Landsat-5.

A partir dos anos 1970, a ocupação pelo homem do espaço do centro-oeste brasileiro apresentou um elevado crescimento devido a políticas de expansão agrícola. Este fato ocorreu por meio do alto grau de mecanização agrícola e aplicação de fertilizantes, visando elevados níveis de produção em diversas localidades, como o sudoeste do Estado de Goiás. Tal predicado da alta produtividade mantém-se até os dias atuais, indicando a grande intensidade da dinâmica de uso e cobertura das terras nesta região. Desta forma, tornase necessário o conhecimento da dinâmica e distribuição espacial dos padrões de uso e cobertura da terra, podendo fornecer subsídios a ações de planejamento agrícola sobre o espaço em alguns municípios do sudoeste goiano. Para isto, imagens orbitais do satélite Landsat TM-5 foram adquiridas em diferentes períodos do ciclo agrícola ao longo de 2007. Informações complementares acerca do uso regional foram utilizadas para apoiar a interpretação e classificação, principalmente a partir dos dados obtidos em campo. Os mapas de uso e cobertura da terra para os municípios de Rio Verde, Acreúna, Santo Antônio da Barra, Santa Helena de Goiás, Montividiu e Paraúna foram obtidos utilizando ferramentas do programa Spring 4.3.3 como a segmentação de imagens, bem como o classificador semi-automático Bhattacharya Distance, sendo estabelecidas dez classes temáticas, com base na legenda proposta pelo IBGE e Corine. A análise multitemporal, assim como a segmentação mostraram-se eficientes na distinção das classes de uso e cobertura da terra da região. A classe de uso destinada ao plantio da soja apresentou o maior percentual da área, mudando para culturas safrinha, solo exposto ou pousio no inverno. Outras classes também merecem destaque como a Pastagem e a Cana-de-açúcar, que apresentaram distribuição espacial bastante concentrada. Este mapeamento fornece subsídios ao planejamento do uso e ocupação das terras na região, considerando os aspectos ambientais e sociais, assegurando maior produtividade agrícola, visando um manejo sustentável das terras e a qualidade de vida ao homem do campo.

Fármacos que têm por alvo o epigenoma

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Structure and denaturation of 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1

The secondary structure of the trimeric protein 4-chlorobenzoyl coenzyme A dehalogenase from Arthrobacter sp. strain TM-1, the second of three enzymes involved in the dechlorination of 4-chlorobenzoate to form 4-hydroxybenzoate, has been examined. E(mM) for the enzyme was 12.59. Analysis by circular dichroism spectrometry in the far uv indicated that 4-chlorobenzoyl coenzyme A dehalogenase was composed mostly of alpha-helix (56%) with lesser amounts of random coil (21%), beta-turn (13%) and beta-sheet (9%). These data are in close agreement with a computational prediction of secondary structure from the primary amino acid sequence, which indicated 55.8% alpha-helix, 33.7% random coil and 10.5% beta-sheet; the enzyme is, therefore, similar to the 4-chlorobenzoyl coenzyme A dehalogenase from Pseudomonas sp. CBS-3. The three-dimensional structure, including that of the presumed active site, predicted by computational analysis, is also closely similar to that of the Pseudomonas dehalogenase. Study of the stability and physicochemical properties revealed that at room temperature, the enzyme was stable for 24 h but was completely inactivated by heating to 60 degrees C for 5 min; thereafter by cooling at 1 degrees C min(-1) to 45 degrees C, 20.6% of the activity could be recovered. Mildly acidic (pH 5.2) or alkaline (pH 10.1) conditions caused complete inactivation, but activity was fully recovered on returning the enzyme to pH 7.4. Circular dichroism studies also indicated that secondary structure was little altered by heating to 60 degrees C, or by changing the pH from 7.4 to 6.0 or 9.2. Complete, irreversible destruction of, and maximal decrease in the fluorescence yield of the protein at 330-350 nm were brought about by 4.5 M urea or 1.1 M guanidinium chloride. Evidence was obtained to support the hypothetical three-dimensional model, that residues W140 and W167 are buried in a non-polar environment, whereas W182 appears at or close to the surface of the protein. At least one of the enzymes of the dehalogenase system (the combined 4-chlorobenzoate:CoA ligase, the dehalogenase and 4-hydroxybenzoyl coenzyme A thioesterase) appears to be capable of association with the cell membrane.

Expression of the 67 kDa laminin receptor (67LR) during retinal development: correlations with angiogenesis

Interaction of vascular cells with the laminin component of basement membranes is important for normal cell function. Likewise, abnormal interactions may have a critical role in vascular pathology. It has been previously demonstrated that the 67 kDa laminin receptor (67LR) is expressed at high levels during proliferative retinopathy in a mouse model and in the current study we have examined 67LR in the neonatal mouse to determine if this receptor plays a role in aspects of developmental angiogenesis in the developing murine retina. Groups of C57/BL6 mice were killed at postnatal day P1, P3, P5, P7, P9 and P11 to assess the retinal vasculature. A number of mice were perfused with FITC-dextran and the eyes removed, fixed in 4% paraformaldehyde (PFA) and flat-mounted for confocal scanning laser microscopy. The eyes from the remaining mice were either placed in 4% PFA and embedded in paraffin-wax, or had the neural retina dissected off and total RNA or protein extracted. Immunofluorescence, in situ hybridization, quantitative reverse transcriptase polymerase chain reaction and Western blotting analysis were employed to locate and determine expression levels of 67LR. Both 67LR mRNA and protein expression showed a characteristic bi-phasic expression pattern which correlated with key stages of retinal vascular development in the murine retina. 67LR showed high expression levels at P1 (P < 0.05) (correlating with superficial vascular plexus formation) and at P7 (P < 0.05) (correlating with deep vascular plexus formation). Conversely, 67LR expression was decreased when active angiogenic activity was lowest. Significantly, optical sectioning of retinal flat-mounts revealed high levels of 67LR expression in developing segments of both superficial and deep capillary plexi, a pattern which co-localized strongly with laminin. 67LR is regulated during post-natal development of the retinal vasculature. High levels of 67LR during the two well-defined phases of retinal capillary plexus formation suggests that this receptor may play an important role in retinal angiogenesis.

Synthetic peptides interacting with the 67-kd laminin receptor can reduce retinal ischemia and inhibit hypoxia-induced retinal neovascularization.

The high-affinity 67-kd laminin receptor (67LR) is expressed by proliferating endothelial cells during retinal neovascularization. The role of 67LR has been further examined experimentally by administration of selective 67LR agonists and antagonists in a murine model of proliferative retinopathy. These synthetic 67LR ligands have been previously shown to stimulate or inhibit endothelial cell motility in vitro without any direct effect on proliferation. In the present study, a fluorescently labeled 67LR antagonist (EGF33–42) was injected intraperitoneally into mice and its distribution in the retina was assessed by confocal scanning laser microscopy. Within 2 hours this peptide was localized to the retinal vasculature, including preretinal neovascular complexes, and a significant amount had crossed the blood retinal barrier. For up to 24 hours postinjection, the peptide was still present in the retinal vascular walls and, to a lesser extent, in the neural retina. Non-labeled EGF33–42 significantly inhibited pre-retinal neovascularization in comparison to controls treated with phosphate-buffered saline or scrambled peptide (P <0.0001). The agonist peptide (Lamß1925–933) also significantly inhibited proliferative retinopathy; however, it caused a concomitant reduction in retinal ischemia in this model by promoting significant revascularization of the central retina (P <0.001). Thus, 67LR appears to be an important target receptor for the modulation of retinal neovascularization. Agonism of this receptor may be valuable in reducing the hypoxia-stimulated release of angiogenic growth factors which drives retinal angiogenesis.

The 67 kDa laminin receptor: structure, function and role in disease

The 67LR (67 kDa laminin receptor) is a cell-surface receptor with high affinity for its primary ligand. Its role as a laminin receptor makes it an important molecule both in cell adhesion to the basement membrane and in signalling transduction following this binding event. The protein also plays critical roles in the metastasis of tumour cells. Isolation of the protein from either normal or cancerous cells results in a product with an approx. molecular mass of 67 kDa. This protein is believed to be derived from a smaller precursor, the 37LRP (37 kDa laminin receptor precursor). However, the precise mechanism by which cytoplasmic 37LRP becomes cell-membrane-embedded 67LR is unclear. The process may involve post-translational fatty acylation of the protein combined with either homo- or hetero-dimerization, possibly with a galectin-3-epitope-containing partner. Furthermore, it has become clear that acting as a receptor for laminin is not the only function of this protein. 67LR also acts as a receptor for viruses, such as Sindbis virus and dengue virus, and is involved with internalization of the prion protein. Interestingly, unmodified 37LRP is a ribosomal component and homologues of this protein are found in all five kingdoms. In addition, it appears to be strongly associated with histones in the eukaryotic cell nucleus, although the precise role of these interactions is not clear. Here we review the current understanding of the structure and function of this molecule, as well as highlighting areas requiring further research.