Leishmania-Specific Surface Antigens Show Sub-Genus Sequence Variation and Immune Recognition

Background: A family of hydrophilic acylated surface (HASP) proteins, containing extensive and variant amino acid repeats, is expressed at the plasma membrane in infective extracellular (metacyclic) and intracellular (amastigote) stages of Old World Leishmania species. While HASPs are antigenic in the host and can induce protective immune responses, the biological functions of these Leishmania-specific proteins remain unresolved. Previous genome analysis has suggested that parasites of the sub-genus Leishmania (Viannia) have lost HASP genes from their genomes. Methods/Principal Findings: We have used molecular and cellular methods to analyse HASP expression in New World Leishmania mexicana complex species and show that, unlike in L. major, these proteins are expressed predominantly following differentiation into amastigotes within macrophages. Further genome analysis has revealed that the L. (Viannia) species, L. (V.) braziliensis, does express HASP-like proteins of low amino acid similarity but with similar biochemical characteristics, from genes present on a region of chromosome 23 that is syntenic with the HASP/SHERP locus in Old World Leishmania species and the L. (L.) mexicana complex. A related gene is also present in Leptomonas seymouri and this may represent the ancestral copy of these Leishmania-genus specific sequences. The L. braziliensis HASP-like proteins (named the orthologous (o) HASPs) are predominantly expressed on the plasma membrane in amastigotes and are recognised by immune sera taken from 4 out of 6 leishmaniasis patients tested in an endemic region of Brazil. Analysis of the repetitive domains of the oHASPs has shown considerable genetic variation in parasite isolates taken from the same patients, suggesting that antigenic change may play a role in immune recognition of this protein family. Conclusions/Significance: These findings confirm that antigenic hydrophilic acylated proteins are expressed from genes in the same chromosomal region in species across the genus Leishmania. These proteins are surface-exposed on amastigotes (although L. (L.) major parasites also express HASPB on the metacyclic plasma membrane). The central repetitive domains of the HASPs are highly variant in their amino acid sequences, both within and between species, consistent with a role in immune recognition in the host.

O contributo da rotina Ler, contar e mostrar para o desenvolvimento de competências da leitura e de expressão oral

Relatório de estágio apresentado à Escola Superior de Educação de Lisboa para obtenção de grau de mestre em Ensino do 1º e 2º ciclo do Ensino Básico

The Inventor Balance and the Functional Specialization in Global Inventive Activities

We study the functional specialization whereby some countries contribute relatively more inventors vs. organizations in the production of inventions at a global scale. We propose a conceptual framework to explain this type of functional specialization, which posits the presence of feedbacks between two distinct sub-systems, each one providing inventors and organizations. We quantify the phenomenon by means of a new metric, the “inventor balance”, which we compute using patent data. We show that the observed imbalances, which are often conspicuous, are determined by several factors: the innovativeness of a country relative to its level of economic development, relative factor endowments, the degree of technological specialization and, last, cultural traits. We argue that the “inventor balance” is a useful indicator for policy makers, and its routine analysis could lead to better informed innovation policies.

Characters Extraction for Traffic Sign Destination boards in video and still images

Traffic Control Signs or destination boards on roadways offer significant information for drivers. Regulation signs tell something like your speed, turns, etc; Warning signs warn drivers of conditions ahead to help them avoid accidents; Destination signs show distances and directions to various locations; Service signs display location of hospitals, gas and rest areas etc. Because the signs are so important and there is always a certain distance from them to drivers, to let the drivers get information clearly and easily even in bad weather or other situations. The idea is to develop software which can collect useful information from a special camera which is mounted in the front of a moving car to extract the important information and finally show it to the drivers. For example, when a frame contains on a destination drive sign board it will be text something like "Linkoping 50",so the software should extract every character of "Linkoping 50", compare them with the already known character data in the database. if there is extracted character match "k" in the database then output the destination name and show to the driver. In this project C++ will be used to write the code for this software.

Efficient parallel proximity queries and an application to highly complex motion planning problems with many narrow passages

In vielen Bereichen der industriellen Fertigung, wie zum Beispiel in der Automobilindustrie, wer- den digitale Versuchsmodelle (sog. digital mock-ups) eingesetzt, um die Entwicklung komplexer Maschinen m ̈oglichst gut durch Computersysteme unterstu ̈tzen zu k ̈onnen. Hierbei spielen Be- wegungsplanungsalgorithmen eine wichtige Rolle, um zu gew ̈ahrleisten, dass diese digitalen Pro- totypen auch kollisionsfrei zusammengesetzt werden k ̈onnen. In den letzten Jahrzehnten haben sich hier sampling-basierte Verfahren besonders bew ̈ahrt. Diese erzeugen eine große Anzahl von zuf ̈alligen Lagen fu ̈r das ein-/auszubauende Objekt und verwenden einen Kollisionserken- nungsmechanismus, um die einzelnen Lagen auf Gu ̈ltigkeit zu u ̈berpru ̈fen. Daher spielt die Kollisionserkennung eine wesentliche Rolle beim Design effizienter Bewegungsplanungsalgorith- men. Eine Schwierigkeit fu ̈r diese Klasse von Planern stellen sogenannte “narrow passages” dar, schmale Passagen also, die immer dort auftreten, wo die Bewegungsfreiheit der zu planenden Objekte stark eingeschr ̈ankt ist. An solchen Stellen kann es schwierig sein, eine ausreichende Anzahl von kollisionsfreien Samples zu finden. Es ist dann m ̈oglicherweise n ̈otig, ausgeklu ̈geltere Techniken einzusetzen, um eine gute Performance der Algorithmen zu erreichen.rnDie vorliegende Arbeit gliedert sich in zwei Teile: Im ersten Teil untersuchen wir parallele Kollisionserkennungsalgorithmen. Da wir auf eine Anwendung bei sampling-basierten Bewe- gungsplanern abzielen, w ̈ahlen wir hier eine Problemstellung, bei der wir stets die selben zwei Objekte, aber in einer großen Anzahl von unterschiedlichen Lagen auf Kollision testen. Wir im- plementieren und vergleichen verschiedene Verfahren, die auf Hu ̈llk ̈operhierarchien (BVHs) und hierarchische Grids als Beschleunigungsstrukturen zuru ̈ckgreifen. Alle beschriebenen Verfahren wurden auf mehreren CPU-Kernen parallelisiert. Daru ̈ber hinaus vergleichen wir verschiedene CUDA Kernels zur Durchfu ̈hrung BVH-basierter Kollisionstests auf der GPU. Neben einer un- terschiedlichen Verteilung der Arbeit auf die parallelen GPU Threads untersuchen wir hier die Auswirkung verschiedener Speicherzugriffsmuster auf die Performance der resultierenden Algo- rithmen. Weiter stellen wir eine Reihe von approximativen Kollisionstests vor, die auf den beschriebenen Verfahren basieren. Wenn eine geringere Genauigkeit der Tests tolerierbar ist, kann so eine weitere Verbesserung der Performance erzielt werden.rnIm zweiten Teil der Arbeit beschreiben wir einen von uns entworfenen parallelen, sampling- basierten Bewegungsplaner zur Behandlung hochkomplexer Probleme mit mehreren “narrow passages”. Das Verfahren arbeitet in zwei Phasen. Die grundlegende Idee ist hierbei, in der er- sten Planungsphase konzeptionell kleinere Fehler zuzulassen, um die Planungseffizienz zu erh ̈ohen und den resultierenden Pfad dann in einer zweiten Phase zu reparieren. Der hierzu in Phase I eingesetzte Planer basiert auf sogenannten Expansive Space Trees. Zus ̈atzlich haben wir den Planer mit einer Freidru ̈ckoperation ausgestattet, die es erlaubt, kleinere Kollisionen aufzul ̈osen und so die Effizienz in Bereichen mit eingeschr ̈ankter Bewegungsfreiheit zu erh ̈ohen. Optional erlaubt unsere Implementierung den Einsatz von approximativen Kollisionstests. Dies setzt die Genauigkeit der ersten Planungsphase weiter herab, fu ̈hrt aber auch zu einer weiteren Perfor- mancesteigerung. Die aus Phase I resultierenden Bewegungspfade sind dann unter Umst ̈anden nicht komplett kollisionsfrei. Um diese Pfade zu reparieren, haben wir einen neuartigen Pla- nungsalgorithmus entworfen, der lokal beschr ̈ankt auf eine kleine Umgebung um den bestehenden Pfad einen neuen, kollisionsfreien Bewegungspfad plant.rnWir haben den beschriebenen Algorithmus mit einer Klasse von neuen, schwierigen Metall- Puzzlen getestet, die zum Teil mehrere “narrow passages” aufweisen. Unseres Wissens nach ist eine Sammlung vergleichbar komplexer Benchmarks nicht ̈offentlich zug ̈anglich und wir fan- den auch keine Beschreibung von vergleichbar komplexen Benchmarks in der Motion-Planning Literatur.

Mice lacking the gene encoding tissue-type plasminogen activator show a selective interference with late-phase long-term potentiation in both Schaffer collateral and mossy fiber pathways.

The gene encoding tissue-type plasminogen activator (t-PA) is an immediate response gene, downstream from CREB-1 and other constitutively expressed transcription factors, which is induced in the hippocampus during the late phase of long-term potentiation (L-LTP). Mice in which the t-PA gene has been ablated (t-PA-/-) showed no gross anatomical, electrophysiological, sensory, or motor abnormalities but manifest a selective reduction in L-LTP in hippocampal slices in both the Schaffer collateral-CA1 and mossy fiber-CA3 pathways. t-PA-/- mice also exhibit reduced potentiation by cAMP analogs and D1/D5 agonists. By contrast, hippocampal-dependent learning and memory were not affected in these mice, whereas performance was impaired on two-way active avoidance, a striatum-dependent task. These results provide genetic evidence that t-PA is a downstream effector gene important for L-LTP and show that modest impairment of L-LTP in CA1 and CA3 does not result in hippocampus-dependent behavioral phenotypes.

Essays on Job Search and Labor Market Dynamics

This dissertation consists of three separate essays on job search and labor market dynamics. In the first essay, “The Impact of Labor Market Conditions on Job Creation: Evidence from Firm Level Data”, I study how much changes in labor market conditions reduce employment fluctuations over the business cycle. Changes in labor market conditions make hiring more expensive during expansions and cheaper during recessions, creating counter-cyclical incentives for job creation. I estimate firm level elasticities of labor demand with respect to changes in labor market conditions, considering two margins: changes in labor market tightness and changes in wages. Using employer-employee matched data from Brazil, I find that all firms are more sensitive to changes in wages rather than labor market tightness, and there is substantial heterogeneity in labor demand elasticity across regions. Based on these results, I demonstrate that changes in labor market conditions reduce the variance of employment growth over the business cycle by 20% in a median region, and this effect is equally driven by changes along each margin. Moreover, I show that the magnitude of the effect of labor market conditions on employment growth can be significantly affected by economic policy. In particular, I document that the rapid growth of the national minimum wages in Brazil in 1997-2010 amplified the impact of the change in labor market conditions during local expansions and diminished this impact during local recessions.

In the second essay, “A Framework for Estimating Persistence of Local Labor

Demand Shocks”, I propose a decomposition which allows me to study the persistence of local labor demand shocks. Persistence of labor demand shocks varies across industries, and the incidence of shocks in a region depends on the regional industrial composition. As a result, less diverse regions are more likely to experience deeper shocks, but not necessarily more long lasting shocks. Building on this idea, I propose a decomposition of local labor demand shocks into idiosyncratic location shocks and nationwide industry shocks and estimate the variance and the persistence of these shocks using the Quarterly Census of Employment and Wages (QCEW) in 1990-2013.

In the third essay, “Conditional Choice Probability Estimation of Continuous- Time Job Search Models”, co-authored with Peter Arcidiacono and Arnaud Maurel, we propose a novel, computationally feasible method of estimating non-stationary job search models. Non-stationary job search models arise in many applications, where policy change can be anticipated by the workers. The most prominent example of such policy is the expiration of unemployment benefits. However, estimating these models still poses a considerable computational challenge, because of the need to solve a differential equation numerically at each step of the optimization routine. We overcome this challenge by adopting conditional choice probability methods, widely used in dynamic discrete choice literature, to job search models and show how the hazard rate out of unemployment and the distribution of the accepted wages, which can be estimated in many datasets, can be used to infer the value of unemployment. We demonstrate how to apply our method by analyzing the effect of the unemployment benefit expiration on duration of unemployment using the data from the Survey of Income and Program Participation (SIPP) in 1996-2007.

The Neuromuscular Activity Of Bothriopsis Bilineata Smaragdina (forest Viper) Venom And Its Toxin Bbil-tx (asp49 Phospholipase A2) On Isolated Mouse Nerve-muscle Preparations.

The presynaptic action of Bothriopsis bilineata smaragdina (forest viper) venom and Bbil-TX, an Asp49 PLA2 from this venom, was examined in detail in mouse phrenic nerve-muscle (PND) preparations in vitro and in a neuroblastoma cell line (SK-N-SH) in order to gain a better insight into the mechanism of action of the venom and associated Asp49 PLA2. In low Ca(2+) solution, venom (3μg/ml) caused a quadriphasic response in PND twitch height whilst at 10μg/ml the venom additionally induced an abrupt and marked initial contracture followed by neuromuscular facilitation, rhythmic oscillations of nerve-evoked twitches, alterations in baseline and progressive blockade. The venom slowed the relaxation phase of muscle twitches. In low Ca(2+), Bbil-TX [210nM (3μg/ml)] caused a progressive increase in PND twitch amplitude but no change in the decay time constant. Venom (10μg/ml) and Bbil-TX (210nM) caused minor changes in the compound action potential (CAP) amplitude recorded from sciatic nerve preparations, with no significant effect on rise time and latency; tetrodotoxin (3.1nM) blocked the CAP at the end of the experiments. In mouse triangularis sterni nerve-muscle (TSn-m) preparations, venom (10μg/ml) and Bbil-TX (210nM) significantly reduced the perineural waveform associated with the outward K(+) current while the amplitude of the inward Na(+) current was not significantly affected. Bbil-TX (210nM) caused a progressive increase in the quantal content of TSn-m preparations maintained in low Ca(2+) solution. Venom (3μg/ml) and toxin (210nM) increased the calcium fluorescence in SK-N-SH neuroblastoma cells loaded with Fluo3 AM and maintained in low or normal Ca(2+) solution. In normal Ca(2+), the increase in fluorescence amplitude was accompanied by irregular and frequent calcium transients. In TSn-m preparations loaded with Fluo4 AM, venom (10μg/ml) caused an immediate increase in intracellular Ca(2+) followed by oscillations in fluorescence and muscle contracture; Bbil-TX did not change the calcium fluorescence in TSn-m preparations. Immunohistochemical analysis of toxin-treated PND preparations revealed labeling of junctional ACh receptors but a loss of the presynaptic proteins synaptophysin and SNAP25. Together, these data confirm the presynaptic action of Bbil-TX and show that it involves modulation of K(+) channel activity and presynaptic protein expression.

Population dynamics: seasonal variation of phytoplankton functional groups in brazilian reservoirs (Billings and Guarapiranga, São Paulo)

Phytoplankton may function as a "sensor" of changes in aquatic environment and responds rapidly to such changes. In freshwaters, coexistence of species that have similar ecological requirements and show the same environmental requirements frequently occurs; such species groups are named functional groups. The use of phytoplankton functional groups to evaluate these changes has proven to be very useful and effective. Thus, the aim of this study was to evaluate the occurrence of functional groups of phytoplankton in two reservoirs (Billings and Guarapiranga) that supply water to millions of people in São Paulo city Metropolitan Area, southeastern Brazil. Surface water samples were collected monthly and physical, chemical and biological (quantitative and qualitative analyses of the phytoplankton) were performed. The highest biovolume (mm³.L-1) of the descriptor species and functional groups were represented respectively by Anabaena circinalis Rabenh. (H1), Microcystis aeruginosa (Kützing) Kützing (L M/M) and Mougeotia sp. (T) in the Guarapiranga reservoir and Cylindrospermopsis raciborskii (Wolosz.) Seen. and Subba Raju (S N), Microcystis aeruginosa and M. panniformis Komárek et al. (L M/M), Planktothrix agardhii (Gom.) Anagn. and Komárek and P. cf. clathrata (Skuja) Anagn. and Komárek (S1) in the Billings reservoir. The environmental factors that most influenced the phytoplankton dynamics were water temperature, euphotic zone, turbidity, conductivity, pH, dissolved oxygen, nitrate and total phosphorous.

Chagas Cardiomyopathy Manifestations and Trypanosoma cruzi Genotypes Circulating in Chronic Chagasic Patients

Chagas disease caused by Trypanosoma cruzi is a complex disease that is endemic and an important problem in public health in Latin America. The T. cruzi parasite is classified into six discrete taxonomic units (DTUs) based on the recently proposed nomenclature (TcI, TcII, TcIII, TcIV, TcV and TcVI). The discovery of genetic variability within TcI showed the presence of five genotypes (Ia, Ib, Ic, Id and Ie) related to the transmission cycle of Chagas disease. In Colombia, TcI is more prevalent but TcII has also been reported, as has mixed infection by both TcI and TcII in the same Chagasic patient. The objectives of this study were to determine the T. cruzi DTUs that are circulating in Colombian chronic Chagasic patients and to obtain more information about the molecular epidemiology of Chagas disease in Colombia. We also assessed the presence of electrocardiographic, radiologic and echocardiographic abnormalities with the purpose of correlating T. cruzi genetic variability and cardiac disease. Molecular characterization was performed in Colombian adult chronic Chagasic patients based on the intergenic region of the mini-exon gene, the 24S alpha and 18S regions of rDNA and the variable region of satellite DNA, whereby the presence of T. cruzi I, II, III and IV was detected. In our population, mixed infections also occurred, with TcI-TcII, TcI-TcIII and TcI-TcIV, as well as the existence of the TcI genotypes showing the presence of genotypes Ia and Id. Patients infected with TcI demonstrated a higher prevalence of cardiac alterations than those infected with TcII. These results corroborate the predominance of TcI in Colombia and show the first report of TcIII and TcIV in Colombian Chagasic patients. Findings also indicate that Chagas cardiomyopathy manifestations are more correlated with TcI than with TcII in Colombia.

Vaccination of Mice with Salmonella Expressing VapA: Mucosal and Systemic Th1 Responses Provide Protection against Rhodococcus equi Infection

Conventional vaccines to prevent the pneumonia caused by Rhodococcus equi have not been successful. We have recently demonstrated that immunization with Salmonella enterica Typhimurium expressing the VapA antigen protects mice against R. equi infection. We now report that oral vaccination of mice with this recombinant strain results in high and persistent fecal levels of antigen-specific IgA, and specific proliferation of the spleen cells of immunized mice in response to the in vitro stimulation with R. equi antigen. After in vitro stimulation, spleen cells of immunized mice produce high levels of Th1 cytokines and show a prominent mRNA expression of the Th1 transcription factor T-bet, in detriment of the Th2 transcription factor GATA-3. Following R. equi challenge, a high H(2)O(2), NO, IL-12, and IFN-gamma content is detected in the organs of immunized mice. On the other hand, TNF-alpha and IL-4 levels are markedly lower in the organs of vaccinated mice, compared with the non-vaccinated ones. The IL-10 content and the mRNA transcription level of TGF-beta are also higher in the organs of immunized mice. A greater incidence of CD4(+) and CD8(+) T cells and B lymphocytes is verified in vaccinated mice. However, there is no difference between vaccinated and non-vaccinated mice in terms of the frequency of CD4(+)CD25(+)Foxp3(+) T cells. Finally, we show that the vaccination confers a long-term protection against R. equi infection. Altogether, these data indicate that the oral vaccination of mice with S. enterica Typhimurium expressing VapA induces specific and long-lasting humoral and cellular responses against the pathogen, which are appropriately regulated and allow tissue integrity after challenge.

NGC 7097: THE ACTIVE GALACTIC NUCLEUS AND ITS MIRROR, REVEALED BY PRINCIPAL COMPONENT ANALYSIS TOMOGRAPHY

Three-dimensional spectroscopy techniques are becoming more and more popular, producing an increasing number of large data cubes. The challenge of extracting information from these cubes requires the development of new techniques for data processing and analysis. We apply the recently developed technique of principal component analysis (PCA) tomography to a data cube from the center of the elliptical galaxy NGC 7097 and show that this technique is effective in decomposing the data into physically interpretable information. We find that the first five principal components of our data are associated with distinct physical characteristics. In particular, we detect a low-ionization nuclear-emitting region (LINER) with a weak broad component in the Balmer lines. Two images of the LINER are present in our data, one seen through a disk of gas and dust, and the other after scattering by free electrons and/or dust particles in the ionization cone. Furthermore, we extract the spectrum of the LINER, decontaminated from stellar and extended nebular emission, using only the technique of PCA tomography. We anticipate that the scattered image has polarized light due to its scattered nature.

Segregation and activation of myosin IIB creates a rear in migrating cells

We have found that MLC-dependent activation of myosin IIB in migrating cells is required to form an extended rear, which coincides with increased directional migration. Activated myosin IIB localizes prominently at the cell rear and produces large, stable actin. lament bundles and adhesions, which locally inhibit protrusion and de. ne the morphology of the tail. Myosin IIA forms de novo. laments away from the myosin IIB-enriched center and back to form regions that support protrusion. The positioning and dynamics of myosin IIA and IIB depend on the self-assembly regions in their coiled-coil C terminus. COS7 and B16 melanoma cells lack myosin IIA and IIB, respectively; and show isoform-specific front-back polarity in migrating cells. These studies demonstrate the role of MLC activation and myosin isoforms in creating a cell rear, the segregation of isoforms during. lament assembly and their differential effects on adhesion and protrusion, and a key role for the noncontractile region of the isoforms in determining their localization and function.

Nuclear modification factors of phi mesons in d plus Au, Cu plus Cu, and Au plus Au collisions at root s(NN)=200 GeV

The PHENIX experiment at the Relativistic Heavy Ion Collider has performed systematic measurements of phi meson production in the K(+)K(-) decay channel at midrapidity in p + p, d + Au, Cu + Cu, and Au + Au collisions at root s(NN) = 200 GeV. Results are presented on the phi invariant yield and the nuclear modification factor R(AA) for Au + Au and Cu + Cu, and R(dA) for d + Au collisions, studied as a function of transverse momentum (1 < p(T) < 7 GeV/c) and centrality. In central and midcentral Au + Au collisions, the R(AA) of phi exhibits a suppression relative to expectations from binary scaled p + p results. The amount of suppression is smaller than that of the pi(0) and the. in the intermediate p(T) range (2-5 GeV/c), whereas, at higher p(T), the phi, pi(0), and. show similar suppression. The baryon (proton and antiproton) excess observed in central Au + Au collisions at intermediate p(T) is not observed for the phi meson despite the similar masses of the proton and the phi. This suggests that the excess is linked to the number of valence quarks in the hadron rather than its mass. The difference gradually disappears with decreasing centrality, and, for peripheral collisions, the R(AA) values for both particle species are consistent with binary scaling. Cu + Cu collisions show the same yield and suppression as Au + Au collisions for the same number of N(part). The R(dA) of phi shows no evidence for cold nuclear effects within uncertainties.

Quantum and pseudoclassical descriptions of nonrelativistic spinning particles in noncommutative space

We construct a nonrelativistic wave equation for spinning particles in the noncommutative space (in a sense, a theta modification of the Pauli equation). To this end, we consider the nonrelativistic limit of the theta-modified Dirac equation. To complete the consideration, we present a pseudoclassical model (a la Berezin-Marinov) for the corresponding nonrelativistic particle in the noncommutative space. To justify the latter model, we demonstrate that its quantization leads to the theta-modified Pauli equation. We extract theta-modified interaction between a nonrelativistic spin and a magnetic field from such a Pauli equation and construct a theta modification of the Heisenberg model for two coupled spins placed in an external magnetic field. In the framework of such a model, we calculate the probability transition between two orthogonal Einstein-Podolsky-Rosen states for a pair of spins in an oscillatory magnetic field and show that some of such transitions, which are forbidden in the commutative space, are possible due to the space noncommutativity. This allows us to estimate an upper bound on the noncommutativity parameter.