Identification of a second homolog of N-ethylmaleimide-sensitive fusion protein that is expressed in the nervous system and secretory tissues of Drosophila.

N-Ethylmaleimide-sensitive fusion protein (NSF) is an ATPase known to have an essential role in intracellular membrane transport events. Recently, cDNA clones encoding a Drosophila melanogaster homolog of this protein, named dNSF, were characterized and found to be expressed in the nervous system. We now report the identification of a second homolog of NSF, called dNSF-2 within this species and report evidence that this ubiquitous and widely utilized fusion protein belongs to a multigene family. The predicted amino acid sequence of dNSF-2 is 84.5% identical to dNSF (hereafter named dNSF-1), 59% identical to NSF from Chinese hamster, and 38.5% identical to the yeast homolog SEC18. The highest similarity was found in a region of dNSF-2 containing one of two ATP-binding sites; this region is most similar to members of a superfamily of ATPases. dNSF-2 is localized to a region between bands 87F12 and 88A3 on chromosome 3, and in situ hybridization techniques revealed expression in the nervous system during embryogenesis and in several imaginal discs and secretory structures in the larvae. Developmental modulation of dNSF-2 expression suggests that quantitative changes in the secretory apparatus are important in histogenesis.

Anti-melanoma antibodies from melanoma patients immunized with genetically modified autologous tumor cells: selection of specific antibodies from single-chain Fv fusion phage libraries.

Fusion phage libraries expressing single-chain Fv antibodies were constructed from the peripheral blood lymphocytes of two melanoma patients who had been immunized with autologous melanoma cells transduced the gamma-interferon gene to enhance immunogenicity, in a trial conducted at another institution. Anti-melanoma antibodies were selected from each library by panning the phage against live cultures of the autologous tumor. After two or three rounds of panning, clones of the phage were tested by ELISA for binding to the autologous tumor cells; > 90% of the clones tested showed a strong ELISA reaction, demonstrating the effectiveness of the panning procedure for selecting antimelanoma antibodies. The panned phage population was extensively absorbed against normal melanocytes to enrich for antibodies that react with melanoma cells but not with melanocytes. The unabsorbed phage were cloned, and the specificities of the expressed antibodies were individually tested by ELISA with a panel of cultured human cells. The first tests were done with normal endothelial and fibroblast cells to identify antibodies that do not react, or react weakly, with two normal cell types, indicating some degree of specificity for melanoma cells. The proportion of phage clones expressing such antibodies was approximately 1%. Those phage were further tested by ELISA with melanocytes, several melanoma lines, and eight other tumor lines, including a glioma line derived from glial cells that share a common lineage with melanocytes. The ELISA tests identified three classes of anti-melanoma antibodies, as follows: (i) a melanoma-specific class that reacts almost exclusively with the melanoma lines; (ii) a tumor-specific class that reacts with melanoma and other tumor lines but does not react with the normal melanocyte, endothelial and fibroblast cells; and (iii) a lineage-specific class that reacts with the melanoma lines, melanocytes, and the glioma line but does not react with the other lines. These are rare classes from the immunized patients' repertoires of anti-melanoma antibodies, most of which are relatively nonspecific anti-self antibodies. The melanoma-specific class was isolated from one patient, and the lineage-specific class was isolated from the other patient, indicating that different patients can have markedly different responses to the same immunization protocol. The procedures described here can be used to screen the antibody repertoire of any person with cancer, providing access to an enormous untapped pool of human monoclonal anti-tumor antibodies with clinical and research potential.

Identification of a plastid protein involved in vesicle fusion and/or membrane protein translocation.

Structural evidence has accumulated suggesting that fusion and/or translocation factors are involved in plastid membrane biogenesis. To test this hypothesis, we have developed an in vitro system in which the extent of fusion and/or translocation is monitored by the conversion of the xanthophyll epoxide (antheraxanthin) into the red ketocarotenoid (capsanthin). Only chromoplast membrane vesicles from red pepper fruits (Capsicum annuum) contain the required enzyme. Vesicles prepared from the mutant yellow cultivar are devoid of this enzyme and accumulate antheraxanthin. The fusion and/or translocation activity is characterized by complementation due to the synthesis of capsanthin and the parallel decrease of antheraxanthin when the two types of vesicles are incubated together in the presence of plastid stroma. We show that the extent of conversion is dependent upon an ATP-requiring protein that is sensitive to N-ethylmaleimide. Further purification and immunological analysis have revealed that the active factor, designated plastid fusion and/or translocation factor (Pftf), resides in a protein of 72 kDa. cDNA cloning revealed that mature Pftf has significant homology to yeast and animal (NSF) or bacterial (Ftsh) proteins involved in vesicle fusion or membrane protein translocation.

Fusion of the TEL gene on 12p13 to the AML1 gene on 21q22 in acute lymphoblastic leukemia.

Chromosomal rearrangements involving band 12p13 are found in a wide variety of human leukemias but are particularly common in childhood acute lymphoblastic leukemia. The genes involved in these rearrangements, however, have not been identified. We now report the cloning of a t(12;21) translocation breakpoint involving 12p13 and 21q22 in two cases of childhood pre-B acute lymphoblastic leukemia, in which t(12;21) rearrangements were not initially apparent. The consequence of the translocation is fusion of the helix-loop-helix domain of TEL, an ETS-like putative transcription factor, to the DNA-binding and transactivation domains of the transcription factor AML1. These data show that TEL, previously shown to be fused to the platelet-derived growth factor receptor beta in chronic myelomonocytic leukemia, can be implicated in the pathogenesis of leukemia through its fusion to either a receptor tyrosine kinase or a transcription factor. The TEL-AML1 fusion also indicates that translocations affecting the AML1 gene can be associated with lymphoid, as well as myeloid, malignancy.

Instabilidades MHD no Tokamak TCABR

Este trabalho descreve o estudo das instabilidades magneto-hidro-dinâmicas (MHD) comumente observadas nas descargas elétricas de plasma no tokamak TCABR, do Instituto de Física da USP. Dois diagnósticos principais foram empregados para observar essas instabilidades: um conjunto poloidal de 24 bobinas magnéticas (bobinas de Mirnov) colocadas próximas à borda do plasma e um medidor de emissões na faixa do Ultra Violeta e de raios X moles com 20 canais (sistema SXR), cujo circuito de condicionamento de sinais foi aprimorado como parte deste trabalho. Esses diagnósticos foram escolhidos porque fornecem informações complementares, uma vez que o sistema SXR observa a parte central da coluna de plasma, enquanto as bobinas de Mirnov detectam as instabilidades MHD na região mais externa da coluna. As informações coletadas por esses diagnósticos foram submetidas à análise espectral com resolução temporal e espacial, possibilitando determinar a evolução das características espectrais e espaciais das instabilidades MHD observadas. Essas análises revelaram que durante a etapa inicial da formação do plasma (quando a corrente de plasma ainda está aumentando) ilhas magnéticas com números de onda decrescente, identificadas como sendo modos kink de borda, são detectadas nas bobinas de Mirnov. Após a formação do plasma, quando os parâmetros de equilíbrio estão relativamente estáveis (platô), oscilações são detectadas tanto nas bobinas de Mirnov quanto no sistema de SXR, indicando a presença de instabilidades MHD em toda a coluna de plasma. Em geral as oscilações medidas nas bobinas de Mirnov tem baixa amplitude e correspondem a pequenas ilhas magnéticas que foram identificadas como sendo modos de ruptura (modos tearing). Por outro lado, as instabilidades na região central foram identificadas como dentes de serra, que correspondem a relaxações periódicas da região interna à superfície magnética com fator de segurança q=1 e que são acompanhadas de oscilações precursoras, cuja amplitude depende da fase do ciclo de relaxação. Devido à essa modulação de amplitude, aparecem picos de frequência satélite nos espectrogramas dos sinais do SXR. Além disso, devido ao fato dos ciclos de relaxação não serem sinusoidais, os harmônicos da frequência de relaxação também aparecem nesses espectrogramas. No entanto, em muitas descargas do TCABR, a intensidade das oscilações medidas nas bobinas de Mirnov aumentam significativamente durante o platô, com efeitos sobre a frequência de todas as instabilidades MHD, até mesmo sobre os dentes de serra localizados na região central da coluna. Em todos os casos, observou-se que durante o platô a frequência das ilhas magnéticas coincide com a frequência das oscilações precursoras do dente de serra, apesar de serem duas instabilidades distintas, localizadas em posições radiais muito diferentes. Essa coincidência de frequências possibilitou descrever a evolução em frequência de todas as oscilações detectadas em diversos diagnósticos com base em apenas duas frequências básicas: a dos ciclos de relaxação dente de serra e a das ilhas magnéticas.

Unifying the observational diversity of isolated neutron stars via magneto-thermal evolution models

Observations of magnetars and some of the high magnetic field pulsars have shown that their thermal luminosity is systematically higher than that of classical radio-pulsars, thus confirming the idea that magnetic fields are involved in their X-ray emission. Here we present the results of 2D simulations of the fully coupled evolution of temperature and magnetic field in neutron stars, including the state-of-the-art kinetic coefficients and, for the first time, the important effect of the Hall term. After gathering and thoroughly re-analysing in a consistent way all the best available data on isolated, thermally emitting neutron stars, we compare our theoretical models to a data sample of 40 sources. We find that our evolutionary models can explain the phenomenological diversity of magnetars, high-B radio-pulsars, and isolated nearby neutron stars by only varying their initial magnetic field, mass and envelope composition. Nearly all sources appear to follow the expectations of the standard theoretical models. Finally, we discuss the expected outburst rates and the evolutionary links between different classes. Our results constitute a major step towards the grand unification of the isolated neutron star zoo.

Portable 3D laser-camera calibration system with color fusion for SLAM

Nowadays, the use of RGB-D sensors have focused a lot of research in computer vision and robotics. These kinds of sensors, like Kinect, allow to obtain 3D data together with color information. However, their working range is limited to less than 10 meters, making them useless in some robotics applications, like outdoor mapping. In these environments, 3D lasers, working in ranges of 20-80 meters, are better. But 3D lasers do not usually provide color information. A simple 2D camera can be used to provide color information to the point cloud, but a calibration process between camera and laser must be done. In this paper we present a portable calibration system to calibrate any traditional camera with a 3D laser in order to assign color information to the 3D points obtained. Thus, we can use laser precision and simultaneously make use of color information. Unlike other techniques that make use of a three-dimensional body of known dimensions in the calibration process, this system is highly portable because it makes use of small catadioptrics that can be placed in a simple manner in the environment. We use our calibration system in a 3D mapping system, including Simultaneous Location and Mapping (SLAM), in order to get a 3D colored map which can be used in different tasks. We show that an additional problem arises: 2D cameras information is different when lighting conditions change. So when we merge 3D point clouds from two different views, several points in a given neighborhood could have different color information. A new method for color fusion is presented, obtaining correct colored maps. The system will be tested by applying it to 3D reconstruction.

Portable 3D laser-camera calibration system with color fusion for SLAM

Paper submitted to the 43rd International Symposium on Robotics (ISR2012), Taipei, Taiwan, Aug. 29-31, 2012.

Plastisol gelation and fusion rheological aspects

This study deals with the rheological aspects of poly-vinyl chloride (PVC) plastisol gelation and fusion processes in foamable formulations. Here, such processes are simulated by temperature-programmed experiment (5 K min−1) in which complex viscosity components are continuously recorded. Nineteen samples based on a PVC-VAC (vinyl acetate 95/5) copolymer with 100 phr plasticizer have been studied, differing only by the plasticizer structure. The sample shear modulus increases continuously with temperature until a maximum, long time after the end of the dissolution process as characterized by DSC. The temperature at the maximum varies between 345 and 428 K with a clear tendency to increase almost linearly with the plasticizer molar mass, and to vary with the flexibility and the degree of branching of the plasticizer molecule. The shear modulus increase is interpreted in terms of progressive “welding” of swelled particles by polymer chain reptation. The plasticizer nature would mainly affect the friction parameter of chain diffusion.

Population synthesis of isolated neutron stars with magneto-rotational evolution

We revisit the population synthesis of isolated radio-pulsars incorporating recent advances on the evolution of the magnetic field and the angle between the magnetic and rotational axes from new simulations of the magneto-thermal evolution and magnetosphere models, respectively. An interesting novelty in our approach is that we do not assume the existence of a death line. We discuss regions in parameter space that are more consistent with the observational data. In particular, we find that any broad distribution of birth spin periods with P0 ≲ 0.5 s can fit the data, and that if the alignment angle is allowed to vary consistently with the torque model, realistic magnetospheric models are favoured compared to models with classical magneto-dipolar radiation losses. Assuming that the initial magnetic field is given by a lognormal distribution, our optimal model has mean strength 〈log B0[G]〉 ≈ 13.0–13.2 with width σ(log B0) = 0.6–0.7. However, there are strong correlations between parameters. This degeneracy in the parameter space can be broken by an independent estimate of the pulsar birth rate or by future studies correlating this information with the population in other observational bands (X-rays and γ-rays).

Population synthesis of isolated neutron stars with magneto-rotational evolution – II. From radio-pulsars to magnetars

Population synthesis studies constitute a powerful method to reconstruct the birth distribution of periods and magnetic fields of the pulsar population. When this method is applied to populations in different wavelengths, it can break the degeneracy in the inferred properties of initial distributions that arises from single-band studies. In this context, we extend previous works to include X-ray thermal emitting pulsars within the same evolutionary model as radio-pulsars. We find that the cumulative distribution of the number of X-ray pulsars can be well reproduced by several models that, simultaneously, reproduce the characteristics of the radio-pulsar distribution. However, even considering the most favourable magneto-thermal evolution models with fast field decay, lognormal distributions of the initial magnetic field overpredict the number of visible sources with periods longer than 12 s. We then show that the problem can be solved with different distributions of magnetic field, such as a truncated lognormal distribution, or a binormal distribution with two distinct populations. We use the observational lack of isolated neutron stars (NSs) with spin periods P > 12 s to establish an upper limit to the fraction of magnetars born with B > 1015 G (less than 1 per cent). As future detections keep increasing the magnetar and high-B pulsar statistics, our approach can be used to establish a severe constraint on the maximum magnetic field at birth of NSs.

Contribution to Real-Time Estimation of Crop Phenological States in a Dynamical Framework Based on NDVI Time Series: Data Fusion With SAR and Temperature

In this study, a methodology based in a dynamical framework is proposed to incorporate additional sources of information to normalized difference vegetation index (NDVI) time series of agricultural observations for a phenological state estimation application. The proposed implementation is based on the particle filter (PF) scheme that is able to integrate multiple sources of data. Moreover, the dynamics-led design is able to conduct real-time (online) estimations, i.e., without requiring to wait until the end of the campaign. The evaluation of the algorithm is performed by estimating the phenological states over a set of rice fields in Seville (SW, Spain). A Landsat-5/7 NDVI series of images is complemented with two distinct sources of information: SAR images from the TerraSAR-X satellite and air temperature information from a ground-based station. An improvement in the overall estimation accuracy is obtained, especially when the time series of NDVI data is incomplete. Evaluations on the sensitivity to different development intervals and on the mitigation of discontinuities of the time series are also addressed in this work, demonstrating the benefits of this data fusion approach based on the dynamic systems.