MP2RAGE, a self bias-field corrected sequence for improved segmentation and T1-mapping at high field.

The large spatial inhomogeneity in transmit B(1) field (B(1)(+)) observable in human MR images at high static magnetic fields (B(0)) severely impairs image quality. To overcome this effect in brain T(1)-weighted images, the MPRAGE sequence was modified to generate two different images at different inversion times, MP2RAGE. By combining the two images in a novel fashion, it was possible to create T(1)-weighted images where the result image was free of proton density contrast, T(2) contrast, reception bias field, and, to first order, transmit field inhomogeneity. MP2RAGE sequence parameters were optimized using Bloch equations to maximize contrast-to-noise ratio per unit of time between brain tissues and minimize the effect of B(1)(+) variations through space. Images of high anatomical quality and excellent brain tissue differentiation suitable for applications such as segmentation and voxel-based morphometry were obtained at 3 and 7 T. From such T(1)-weighted images, acquired within 12 min, high-resolution 3D T(1) maps were routinely calculated at 7 T with sub-millimeter voxel resolution (0.65-0.85 mm isotropic). T(1) maps were validated in phantom experiments. In humans, the T(1) values obtained at 7 T were 1.15+/-0.06 s for white matter (WM) and 1.92+/-0.16 s for grey matter (GM), in good agreement with literature values obtained at lower spatial resolution. At 3 T, where whole-brain acquisitions with 1 mm isotropic voxels were acquired in 8 min, the T(1) values obtained (0.81+/-0.03 s for WM and 1.35+/-0.05 for GM) were once again found to be in very good agreement with values in the literature.

Adherence as a predictor of the development of class-specific resistance mutations: the Swiss HIV Cohort Study.

BACKGROUND: Non-adherence is one of the strongest predictors of therapeutic failure in HIV-positive patients. Virologic failure with subsequent emergence of resistance reduces future treatment options and long-term clinical success. METHODS: Prospective observational cohort study including patients starting new class of antiretroviral therapy (ART) between 2003 and 2010. Participants were naïve to ART class and completed ≥1 adherence questionnaire prior to resistance testing. Outcomes were development of any IAS-USA, class-specific, or M184V mutations. Associations between adherence and resistance were estimated using logistic regression models stratified by ART class. RESULTS: Of 314 included individuals, 162 started NNRTI and 152 a PI/r regimen. Adherence was similar between groups with 85% reporting adherence ≥95%. Number of new mutations increased with increasing non-adherence. In NNRTI group, multivariable models indicated a significant linear association in odds of developing IAS-USA (odds ratio (OR) 1.66, 95% confidence interval (CI): 1.04-2.67) or class-specific (OR 1.65, 95% CI: 1.00-2.70) mutations. Levels of drug resistance were considerably lower in PI/r group and adherence was only significantly associated with M184V mutations (OR 8.38, 95% CI: 1.26-55.70). Adherence was significantly associated with HIV RNA in PI/r but not NNRTI regimens. CONCLUSION: Therapies containing PI/r appear more forgiving to incomplete adherence compared with NNRTI regimens, which allow higher levels of resistance, even with adherence above 95%. However, in failing PI/r regimens good adherence may prevent accumulation of further resistance mutations and therefore help to preserve future drug options. In contrast, adherence levels have little impact on NNRTI treatments once the first mutations have emerged.

Identification of naturally processed hepatitis C virus-derived major histocompatibility complex class I ligands.

Fine mapping of human cytotoxic T lymphocyte (CTL) responses against hepatitis C virus (HCV) is based on external loading of target cells with synthetic peptides which are either derived from prediction algorithms or from overlapping peptide libraries. These strategies do not address putative host and viral mechanisms which may alter processing as well as presentation of CTL epitopes. Therefore, the aim of this proof-of-concept study was to identify naturally processed HCV-derived major histocompatibility complex (MHC) class I ligands. To this end, continuous human cell lines were engineered to inducibly express HCV proteins and to constitutively express high levels of functional HLA-A2. These cell lines were recognized in an HLA-A2-restricted manner by HCV-specific CTLs. Ligands eluted from HLA-A2 molecules isolated from large-scale cultures of these cell lines were separated by high performance liquid chromatography and further analyzed by electrospray ionization quadrupole time of flight mass spectrometry (MS)/tandem MS. These analyses allowed the identification of two HLA-A2-restricted epitopes derived from HCV nonstructural proteins (NS) 3 and 5B (NS3₁₄₀₆₋₁₄₁₅ and NS5B₂₅₉₄₋₂₆₀₂). In conclusion, we describe a general strategy that may be useful to investigate HCV pathogenesis and may contribute to the development of preventive and therapeutic vaccines in the future.

Purification and ligand binding of a soluble class I major histocompatibility complex molecule consisting of the first three domains of H-2Kd fused to beta 2-microglobulin expressed in the baculovirus-insect cell system.

A recombinant baculovirus encoding a single-chain murine major histocompatibility complex class I molecule in which the first three domains of H-2Kd are fused to beta 2-microglobulin (beta 2-m) via a 15-amino acid linker has been isolated and used to infect lepidopteran cells. A soluble, 391-amino acid single-chain H-2Kd (SC-Kd) molecule of 48 kDa was synthesized and glycosylated in insect cells and could be purified in the absence of detergents by affinity chromatography using the anti-H-2Kd monoclonal antibody SF1.1.1.1. We tested the ability of SC-Kd to bind antigenic peptides using a direct binding assay based on photoaffinity labeling. The photoreactive derivative was prepared from the H-2Kd-restricted Plasmodium berghei circumsporozoite protein (P.b. CS) peptide 253-260 (YIPSAEKI), a probe that we had previously shown to be unable to bind to the H-2Kd heavy chain in infected cells in the absence of co-expressed beta 2-microglobulin. SC-Kd expressed in insect cells did not require additional mouse beta 2-m to bind the photoprobe, indicating that the covalently attached beta 2-m could substitute for the free molecule. Similarly, binding of the P.b. CS photoaffinity probe to the purified SC-Kd molecule was unaffected by the addition of exogenous beta 2-m. This is in contrast to H-2KdQ10, a soluble H-2Kd molecule in which beta 2-m is noncovalently bound to the soluble heavy chain, whose ability to bind the photoaffinity probe is greatly enhanced in the presence of an excess of exogenous beta 2-m. The binding of the probe to SC-Kd was allele-specific, since labeling was selectively inhibited only by antigenic peptides known to be presented by the H-2Kd molecule.

Aplicação de SIG no Processamento do Inventário Florestal Nacional – Cabo Verde

Cabo Verde desde do século passado tem envidado esforço na florestação, sobretudo depois de 1975 para atenuar os efeitos da seca e da desertificação criando deste modo grandes áreas arborizadas. Entretanto, à medida que os recursos florestais foram sendo criados, a problemática da sua avaliação e da sua gestão sustentável, passaram a merecer maior atenção das autoridades nacionais. A lei florestal, promulgada em 1998 define como uma das atribuições e acções do Estado, através dos serviços florestais, a elaboração dos planos de gestão das zonas florestais. Este plano de gestão implica a análise e a apreciação de dados concretos e actualizados sobre a situação real das zonas florestais, sendo possível apenas através do inventário florestal nacional (IFN). Neste trabalho é proposta uma metodologia de processamento do IFN em que se utilizam as potencialidades dos Sistemas de Informação Geográfica (SIG). Foram utilizados para este trabalho os programas: ArcGis 9.1, para produção cartográfica, geoprocessamento e análise espacial e o Field-Map 8.1 para a classificação de ortofotos num esquema de classificação hierárquica, em cinco níveis, adaptado a Cabo Verde (classes de uso do solo adoptado ao esquema de classificação do território europeu – CORINE Land Cover e da Organização das Nações Unidas para a Agricultura e Alimentação (FAO). Os dados utilizados foram compilados no âmbito do projecto do inventário florestal. Os resultados obtidos, para a Ilha de Santiago, constituem uma base cartográfica para o IFN com diversos temas cartográficos, nomeadamente, mapas das zonas florestadas, mapas de ocupação do solo e mapas de amostras inventariáveis cuja metodologia de elaboração poderá ser facilmente replicada para as restantes ilhas do arquipélago.

Distribuição Espacial da Susceptibilidade à Erosão Hídrica nas Bacias das Ribeiras de Picos e Seca (Santiago, Cabo Verde)

Os processos de erosão hídrica em Cabo Verde são os mais marcantes da dinâmica actual das vertentes, pois são os mais comuns e que afectam áreas extensasdurante a curta estação húmida de três meses. A ocorrência de episódios chuvosos concentrados no tempo e com uma evidente irregularidade espacial permitem umaacentuada erosividade das precipitações, marcada por uma forte irregularidade regional. A forte variabilidade das formas de relevo, a diversidade da natureza das unidadesgeológicas e a multiplicidade de ocupação do solo favorecem condições deerodibilidade muito contrastadas no espaço. O objectivo deste trabalho é estabelecer um modelo desusceptibilidade à erosão hídricaem função de factores geomorfológicos (declive, perfil e traçado das vertentes eerodibilidade das unidades litológicas e dos materiais de cobertura), climáticos(intensidade pluviométrica) e de ocupação do solo para as bacias das ribeiras dos Picose Seca. Os resultados foram obtidos com recurso ao ambiente de Sistemas deInformação Geográfica (SIG). Este trabalho surge na sequência de outros já realizadospelos autores, onde se apresentaram as condições de erodibilidade e erosividade paraáreas mais restritas da Ilha de Santiago. O modelo de susceptibilidade à erosão hídrica resultou do cruzamento dos mapas dedeclives, de perfil e do traçado das vertentes, obtidos a partir do modelo digital deterreno (DTM), do mapa geológico, da distribuição espacial da intensidadepluviométrica e da densidade de ocupação do solo, tendo em conta que são estas asprincipais condicionantes de erosão hídrica, referidas pelos autores que estudaram estaregião. Cada um destes mapas foi reclassificado com base numa análise qualitativa dograu de erodibilidade, sendo atribuído um número de ordem a cada classe, em função da sua susceptibilidade à erosão hídrica, conforme foi localmente reconhecido. Verifica-se que as áreas de maior susceptibilidade à erosão hídrica são as do sectorsudeste da bacia da Ribeira Seca e as vertentes dos principais vales da bacia da Ribeira dos Picos, onde se encontram as unidades geológicas mais friáveis, os declives mais acentuados e onde predominam sectores das vertentes de traçado côncavo, a que seassocia pontualmente a mais elevada intensidade pluviométrica.

Increasing Students Vocabulary in Efl Class

After reviewing the literature, this work tries to show the importance of teaching vocabulary fõr students’ literacy skills, especially, reading comprehension. Many researchers suggest that the greatest amount of vocabulary growth occurs through incidental word learning in wide reading, and, research indicates that vocabulary instruction is an important vehicle for vocabulary learning. (Anderson& Nagy, as cited in Harmon, 1992, p.306). Word knowledge is one of the best ways of successful reading and comprehension. “Reading enhancement correlates with reader’s vocabulary” (Im, 1994, p.12). Therefore, today’s language teachers and researchers have realized the important role of vocabulary in reading comprehension. A survey carried out on 10th, 11th and 12th grade students, regarding their reading comprehension, shows that unknown words is one of the factors which influences their ability to read and comprehend a passage. It also shows that students feel the need to be instructed on strategy when encountering new words and consequently improving their vocabulary. This inhibits their understanding of a reading selection. As a result it is crucial that teachers equip students with methodological tools to be employed when they encounter unknown words. There are a lot systematic approaches for discerning which skills and words a teacher should focus on and meaningful classroom activities to reinforce the words and strategies that teachers can use to help students increase their word knowledge. Finally research indicates that developing students’ vocabulary correlates with success in all areas of curriculum (Edger, 1999, p.14). The success of vocabulary development depends on students’ active process of learning and strategies used by teachers.

A general class of adaptative strategies

We exhibit and characterize an entire class of simple adaptive strategies,in the repeated play of a game, having the Hannan-consistency property: In the long-run, the player is guaranteed an average payoff as large as the best-reply payoff to the empirical distribution of play of the otherplayers; i.e., there is no "regret." Smooth fictitious play (Fudenberg and Levine [1995]) and regret-matching (Hart and Mas-Colell [1998]) areparticular cases. The motivation and application of this work come from the study of procedures whose empirical distribution of play is, in thelong-run, (almost) a correlated equilibrium. The basic tool for the analysis is a generalization of Blackwell's [1956a] approachability strategy for games with vector payoffs.

Finite horizon bargaining and the consistent field

This paper explores the relationships between noncooperative bargaining games and the consistent value for non-transferable utility (NTU) cooperative games. A dynamic approach to the consistent value for NTU games is introduced: the consistent vector field. The main contribution of the paper is to show that the consistent field is intimately related to the concept of subgame perfection for finite horizon noncooperative bargaining games, as the horizon goes to infinity and the cost of delay goes to zero. The solutions of the dynamic system associated to the consistent field characterize the subgame perfect equilibrium payoffs of the noncooperative bargaining games. We show that for transferable utility, hyperplane and pure bargaining games, the dynamics of the consistent fields converge globally to the unique consistent value. However, in the general NTU case, the dynamics of the consistent field can be complex. An example is constructed where the consistent field has cyclic solutions; moreover, the finite horizon subgame perfect equilibria do not approach the consistent value.

In vivo targeting of an anti-tumor antibody coupled to antigenic MHC class I complexes induces specific growth inhibition and regression of established syngeneic tumor grafts.

The concept of antibody-mediated targeting of antigenic MHC/peptide complexes on tumor cells in order to sensitize them to T-lymphocyte cytotoxicity represents an attractive new immunotherapy strategy. In vitro experiments have shown that an antibody chemically conjugated or fused to monomeric MHC/peptide can be oligomerized on the surface of tumor cells, rendering them susceptible to efficient lysis by MHC-peptide restricted specific T-cell clones. However, this strategy has not yet been tested entirely in vivo in immunocompetent animals. To this aim, we took advantage of OT-1 mice which have a transgenic T-cell receptor specific for the ovalbumin (ova) immunodominant peptide (257-264) expressed in the context of the MHC class I H-2K(b). We prepared and characterized conjugates between the Fab' fragment from a high-affinity monoclonal antibody to carcinoembryonic antigen (CEA) and the H-2K(b) /ova peptide complex. First, we showed in OT-1 mice that the grafting and growth of a syngeneic colon carcinoma line transfected with CEA could be specifically inhibited by systemic injections of the conjugate. Next, using CEA transgenic C57BL/6 mice adoptively transferred with OT-1 spleen cells and immunized with ovalbumin, we demonstrated that systemic injections of the anti-CEA-H-2K(b) /ova conjugate could induce specific growth inhibition and regression of well-established, palpable subcutaneous grafts from the syngeneic CEA-transfected colon carcinoma line. These results, obtained in a well-characterized syngeneic carcinoma model, demonstrate that the antibody-MHC/peptide strategy can function in vivo. Further preclinical experimental studies, using an anti-viral T-cell response, will be performed before this new form of immunotherapy can be considered for clinical use.

Staying With the Tension: “Estranged Intimacy” and Gendered Expectations in the Field

The position of the anthropologist in the field is discussed, in this article, as a position of “estranged intimacy”, that is to say, the anthropologist occupies an ambiguous position of becoming intimately involved whilst concurrently standing back. This definition derives from reflections upon fieldwork, conducted in the north of Portugal, with Cape Verdean migrant young women and their experiences as mothers. The article discusses two aspects related to the fieldwork. Firstly, the way in which diverse strategies of establishing relations in the field placed me in a position of “estranged intimacy” which reconfigured the meanings I had initially attributed to the term “Cape Verdean women”. Secondly, how becoming unexpectedly involved in a situation of intense conjugal conflict led me to reconsider my understanding of Cape Verdean gender relations. Both cases demonstrate how the endeavour to produce analytical and ethnographical knowledge was shot through with an unstable mix of detachment and involvement and how coming up against the unexpected may contribute towards the reconfiguration of ethnographic knowledge, in this specific case, with regard to the dynamics of gender relations.

An in vivo ultrahigh field 14.1 T (1) H-MRS study on 6-OHDA and α-synuclein-based rat models of Parkinson's disease: GABA as an early disease marker.

The detection of Parkinson's disease (PD) in its preclinical stages prior to outright neurodegeneration is essential to the development of neuroprotective therapies and could reduce the number of misdiagnosed patients. However, early diagnosis is currently hampered by lack of reliable biomarkers. (1) H magnetic resonance spectroscopy (MRS) offers a noninvasive measure of brain metabolite levels that allows the identification of such potential biomarkers. This study aimed at using MRS on an ultrahigh field 14.1 T magnet to explore the striatal metabolic changes occurring in two different rat models of the disease. Rats lesioned by the injection of 6-hydroxydopamine (6-OHDA) in the medial-forebrain bundle were used to model a complete nigrostriatal lesion while a genetic model based on the nigral injection of an adeno-associated viral (AAV) vector coding for the human α-synuclein was used to model a progressive neurodegeneration and dopaminergic neuron dysfunction, thereby replicating conditions closer to early pathological stages of PD. MRS measurements in the striatum of the 6-OHDA rats revealed significant decreases in glutamate and N-acetyl-aspartate levels and a significant increase in GABA level in the ipsilateral hemisphere compared with the contralateral one, while the αSyn overexpressing rats showed a significant increase in the GABA striatal level only. Therefore, we conclude that MRS measurements of striatal GABA levels could allow for the detection of early nigrostriatal defects prior to outright neurodegeneration and, as such, offers great potential as a sensitive biomarker of presymptomatic PD.