Insulin-like growth factor binding protein 2 (IGFBP2) promotes glioma development and progression

Overexpression of insulin-like growth factor binding protein 2 (IGFBP2) is associated with progression and poor survival in many types of human cancer (such as prostate, ovarian, adrenocortical, breast, colorectal carcinomas, leukemia, and high-grade gliomas). We therefore hypothesize that IGFBP2 is a key regulator of tumor progression. We tested our hypothesis in gliomas using the somatic gene transfer RCAS-tva mouse model system, which permits the introduction of specific genes into specific, cell lineages, in this case glial cells (RCAS: Replication competent avian sarcomavirus, tv-a: avian RCAS virus receptor). Mice are transgenic and harbor the tv-a receptor under the control of a glial-specific promoter and study genes are cloned into the RCAS vector for post-natal intracranial delivery. For these experiments, the study genes were IGFBP2, platelet-derived growth factor B (PDGFB), K-Ras, Akt, and IIp45 (invasion inhibitory protein 45 kDa; known to bind and block IGFBP2 activity), which were delivered separately and in combination. Our results show that PDGFB signaling leads exclusively to the formation of low-grade (WHO grade II) oligodendrogliomas. PDGFB delivered in combination with IGFBP2 results in the formation of anaplastic oligodendrogliomas (WHO grade III), which are characterized by increased cellularity, vascular proliferation, small regions of necrosis, increased mitotic activity, and increased activation of the Akt pathway. IIp45 injected in combination with PDGFB and IGFBP2 ablates IGFBP2-induced tumor progression, which results in formation of low-grade oligodendrogliomas, and an overall reduction in tumor incidence. K-Ras expression was required to form astrocytomas with either IGFBP2 or Akt, indicating the activation of two separate pathways is necessary for gliomagenesis. In ex vivo experiments, blockade of Akt by an inhibitor led to decreased viability of cells co-expressing IGFBP2 versus PDGFB expression alone. This study provides definitive evidence, for the first time, that: (1) IGFBP2 plays a role in activation of the Akt pathway, (2) IGFBP2 collaborates with K-Ras or PDGFB in the development and progression of two major types of glioma, and (3) IGFBP2-induced tumor progression can be ablated by IIp45 or by specific inhibition of the Akt pathway. ^

Comparisson of hydrogen applications for storage and energy vector

For the decades to come can be foreseen that electricity and water will keep be playing a key role in the countries development, both can be considered the most important energy vectors and its control can be crucial for governments, companies and leaders in general. Energy is essential for all human activities and its availability is critical to economic and social development. In particular, electricity, a form of energy, is required to produce goods, to provide medical assistance and basic civic services in education, to assure availability of clean water, to create conducive environment for prosperity and improvement, and to keep an acceptable quality of life. The way in which electricity is generated from different resources varies through the different countries. Nuclear energy controlled within reactors to steam production, gas, fuel-oil and coal fired in power stations, water, solar and wind energy among others are employed, sometimes not very efficiently, to produce electricity. The so call energy mix of an individual country is formed up by the contribution of each resource or form of energy to the electricity generation market of the so country. During the last decade the establishment of proper energy mixes for countries has gained much importance, and energy drivers should enforce long term plans and policies. Hints, reports and guides giving tracks on energy resources contribution are been developed by noticeable organisations like the IEA (International Energy Agency) or the IAEA (International Atomic Energy Agency) and the WEC (World Energy Council). This paper evaluates energy issues the market and countries are facing today regarding energy mix scheduling and panorama. This paper revises and seeks to improve methodology available that are applicable on energy mix plan definition. Key Factors are identified, established and assessed through this paper for the common implementation, the themes driving the future energy mix methodology proposal. Those have a clear influence and are closely related to future environmental policies. Key Factors take into consideration sustainability, energy security, social and economic growth, climate change, air quality and social stability. The strength of the Key Factors application on energy system planning to different countries is contingent on country resources, location, electricity demand and electricity generation industry, technology available, economic situation and prospects, energy policy and regulation

Implementación de una Support Vector Machine en RVC – CAL para imágenes hiperespectrales

El análisis de imágenes hiperespectrales permite obtener información con una gran resolución espectral: cientos de bandas repartidas desde el espectro infrarrojo hasta el ultravioleta. El uso de dichas imágenes está teniendo un gran impacto en el campo de la medicina y, en concreto, destaca su utilización en la detección de distintos tipos de cáncer. Dentro de este campo, uno de los principales problemas que existen actualmente es el análisis de dichas imágenes en tiempo real ya que, debido al gran volumen de datos que componen estas imágenes, la capacidad de cómputo requerida es muy elevada. Una de las principales líneas de investigación acerca de la reducción de dicho tiempo de procesado se basa en la idea de repartir su análisis en diversos núcleos trabajando en paralelo. En relación a esta línea de investigación, en el presente trabajo se desarrolla una librería para el lenguaje RVC – CAL – lenguaje que está especialmente pensado para aplicaciones multimedia y que permite realizar la paralelización de una manera intuitiva – donde se recogen las funciones necesarias para implementar el clasificador conocido como Support Vector Machine – SVM. Cabe mencionar que este trabajo complementa el realizado en [1] y [2] donde se desarrollaron las funciones necesarias para implementar una cadena de procesado que utiliza el método unmixing para procesar la imagen hiperespectral. En concreto, este trabajo se encuentra dividido en varias partes. La primera de ellas expone razonadamente los motivos que han llevado a comenzar este Trabajo de Investigación y los objetivos que se pretenden conseguir con él. Tras esto, se hace un amplio estudio del estado del arte actual y, en él, se explican tanto las imágenes hiperespectrales como sus métodos de procesado y, en concreto, se detallará el método que utiliza el clasificador SVM. Una vez expuesta la base teórica, nos centraremos en la explicación del método seguido para convertir una versión en Matlab del clasificador SVM optimizado para analizar imágenes hiperespectrales; un punto importante en este apartado es que se desarrolla la versión secuencial del algoritmo y se asientan las bases para una futura paralelización del clasificador. Tras explicar el método utilizado, se exponen los resultados obtenidos primero comparando ambas versiones y, posteriormente, analizando por etapas la versión adaptada al lenguaje RVC – CAL. Por último, se aportan una serie de conclusiones obtenidas tras analizar las dos versiones del clasificador SVM en cuanto a bondad de resultados y tiempos de procesado y se proponen una serie de posibles líneas de actuación futuras relacionadas con dichos resultados. ABSTRACT. Hyperspectral imaging allows us to collect high resolution spectral information: hundred of bands covering from infrared to ultraviolet spectrum. These images have had strong repercussions in the medical field; in particular, we must highlight its use in cancer detection. In this field, the main problem we have to deal with is the real time analysis, because these images have a great data volume and they require a high computational power. One of the main research lines that deals with this problem is related with the analysis of these images using several cores working at the same time. According to this investigation line, this document describes the development of a RVC – CAL library – this language has been widely used for working with multimedia applications and allows an optimized system parallelization –, which joins all the functions needed to implement the Support Vector Machine – SVM - classifier. This research complements the research conducted in [1] and [2] where the necessary functions to implement the unmixing method to analyze hyperspectral images were developed. The document is divided in several chapters. The first of them introduces the motivation of the Master Thesis and the main objectives to achieve. After that, we study the state of the art of some technologies related with this work, like hyperspectral images, their processing methods and, concretely, the SVM classifier. Once we have exposed the theoretical bases, we will explain the followed methodology to translate a Matlab version of the SVM classifier optimized to process an hyperspectral image to RVC – CAL language; one of the most important issues in this chapter is that a sequential implementation is developed and the bases of a future parallelization of the SVM classifier are set. At this point, we will expose the results obtained in the comparative between versions and then, the results of the different steps that compose the SVM in its RVC – CAL version. Finally, we will extract some conclusions related with algorithm behavior and time processing. In the same way, we propose some future research lines according to the results obtained in this document.

Three members of a novel small gene-family from Arabidopsis thaliana able to complement functionally an Escherichia coli mutant defective in PAPS reductase activity encode proteins with a thioredoxin-like domain and “APS reductase” activity

Three different cDNAs, Prh-19, Prh-26, and Prh-43 [3′-phosphoadenosine-5′-phosphosulfate (PAPS) reductase homolog], have been isolated by complementation of an Escherichia coli cysH mutant, defective in PAPS reductase activity, to prototrophy with an Arabidopsis thaliana cDNA library in the expression vector λYES. Sequence analysis of the cDNAs revealed continuous open reading frames encoding polypeptides of 465, 458, and 453 amino acids, with calculated molecular masses of 51.3, 50.5, and 50.4 kDa, respectively, that have strong homology with fungal, yeast, and bacterial PAPS reductases. However, unlike microbial PAPS reductases, each PRH protein has an N-terminal extension, characteristic of a plastid transit peptide, and a C-terminal extension that has amino acid and deduced three-dimensional homology to thioredoxin proteins. Adenosine 5′-phosphosulfate (APS) was shown to be a much more efficient substrate than PAPS when the activity of the PRH proteins was tested by their ability to convert 35S-labeled substrate to acid-volatile 35S-sulfite. We speculate that the thioredoxin-like domain is involved in catalytic function, and that the PRH proteins may function as novel “APS reductase” enzymes. Southern hybridization analysis showed the presence of a small multigene family in the Arabidopsis genome. RNA blot hybridization with gene-specific probes revealed for each gene the presence of a transcript of ≈1.85 kb in leaves, stems, and roots that increased on sulfate starvation. To our knowledge, this is the first report of the cloning and characterization of plant genes that encode proteins with APS reductase activity and supports the suggestion that APS can be utilized directly, without activation to PAPS, as an intermediary substrate in reductive sulfate assimilation.

A serotonin transporter gene intron 2 polymorphic region, correlated with affective disorders, has allele-dependent differential enhancer-like properties in the mouse embryo

Polymorphic regions consisting of a variable number of tandem repeats within intron 2 of the gene coding for the serotonin transporter protein 5-HTT have been associated with susceptibility to affective disorders. We have cloned two of these intronic polymorphisms, Stin2.10 and Stin2.12, into an expression vector containing a heterologous minimal promoter and the bacterial LacZ reporter gene. These constructs were then used to produce transgenic mice. In embryonic day 10.5 embryos, both Stin2.10 and Stin2.12 produced consistent β-galactosidase expression in the embryonic midbrain, hindbrain, and spinal cord floor plate. However, we observed that the levels of β-galactosidase expression produced by both the Stin2.10 and Stin2.12 within the rostral hindbrain differed significantly at embryonic day 10.5. Our data suggest that these polymorphic variable number of tandem repeats regions act as transcriptional regulators and have allele-dependent differential enhancer-like properties within an area of the hindbrain where the 5-HTT gene is known to be transcribed at this stage of development.

Antisense RNA to the type I insulin-like growth factor receptor suppresses tumor growth and prevents invasion by rat prostate cancer cells in vivo.

Prostate carcinoma is the second leading cause of death from malignancy in men in the United States. Prostate cancer cells express type I insulin-like growth factor receptor (IGF-IR) and prostate cancer selectively metastazises to bone, which is an environment rich in insulin-like growth factors (IGFs), thereby supporting a paracrine action for cancer cell proliferation. We asked whether the IGF-IR is coupled to tumorigenicity and invasion of prostate cancer. When rat prostate adenocarcinoma cells (PA-III) were stably transfected with an antisense IGF-IR expression construct containing the ZnSO4-inducible metallothionein-1 transcriptional promoter, the transfectants expressed high levels of IGF-IR antisense RNA after induction with ZnSO4, which resulted in dramatically reduced levels of endogenous IGF-IR mRNA. A significant reduction in expression both of tissue-type plasminogen activator and of urokinase-type plasminogen activator occurred in PA-III cells accompanying inhibition of IGF-IR. Subcutaneous injection of either nontransfected PA-III or PA-III cells transfected with vector minus the IGF-IR insert into nude mice resulted in large tumors after 4 weeks. However, mice injected with IGF-IR antisense-transfected PA-III cells either developed tumors 90% smaller than controls or remained tumor-free after 60 days of observation. When control-transfected PA-III cells were inoculated over the abraded calvaria of nude mice, large tumors formed with invasion of tumor cells into the brain parenchyma. In contrast, IGF-IR antisense transfectants formed significantly smaller tumors with no infiltration into brain. These results indicate an important role for the IGF/IGF-IR pathway in metastasis and provide a basis for targeting IGF-IR as a potential treatment for prostate cancer.

Cloning and expression of a gene encoding an integrin-like protein in Candida albicans.

The existence of integrin-like proteins in Candida albicans has been postulated because monoclonal antibodies to the leukocyte integrins alpha M and alpha X bind to blastospores and germ tubes, recognize a candidal surface protein of approximately 185 kDa, and inhibit candidal adhesion to human epithelium. The gene alpha INT1 was isolated from a library of C. albicans genomic DNA by screening with a cDNA probe from the transmembrane domain of human alpha M. The predicted polypeptide (alpha Int1p) of 188 kDa contains several motifs common to alpha M and alpha X: a putative I domain, two EF-hand divalent cation-binding sites, a transmembrane domain, and a cytoplasmic tail with a single tyrosine residue. An internal RGD tripeptide is also present. Binding of anti-peptide antibodies raised to potential extracellular domains of alpha Int1p confirms surface localization in C. albicans blastopores. By Southern blotting, alpha INT1 is unique to C. albicans. Expression of alpha INT1 under control of a galactose-inducible promoter led to the production of germ tubes in haploid Saccharomyces cerevisiae and in the corresponding ste12 mutant. Germ tubes were not observed in haploid yeast transformed with vector alone, in transformants expressing a galactose-inducible gene from Chlamydomonas, or in transformants grown in the presence of glucose or raffinose. Transformants producing alpha Int1p bound an anti-alpha M monoclonal antibody and exhibited enhanced aggregation. Studies of alpha Int1p reveal novel roles for primitive integrin-like proteins in adhesion and in STE12-independent morphogenesis.

Expression of a flower-specific Myb protein in leaf cells using a viral vector causes ectopic activation of a target promoter.

The promoter of the bean PAL2 gene (encoding phenylalanine ammonia-lyase; EC 4.3.1.5) is a model for studies of tissue-restricted gene expression in plants. Petal epidermis is one of the tissues in which this promoter is activated in tobacco. Previous work suggested that a major factor establishing the pattern of PAL2 expression in tobacco petals is the tissue distribution of a protein closely related to Myb305, which is a Myb-like transcriptional activator from snapdragon. In the present work, we show that Myb305 expression in tobacco leaves causes ectopic activation of the PAL2 promoter. To achieve Myb305 expression in planta, a viral expression vector was used. This approach combines the utility of transient assays with the possibility of direct biochemical detection of the introduced factor and may have wider application for studying the function of plant transcription factors.

Interactions of maize bushy stunt phytoplasma with the leafhopper vector, Dalbulus maidis (Delong and Wolcott) (Hemiptera: Cicadellidae) and associated microbiota

Phytoplasmas are bacteria with a persistent propagative transmission by insect vectors that generates direct and indirect interactions among them. In order to understand these interactions for maize bushy stunt phytoplasma (MBSP) and the leafhopper vector Dalbulus maidis (Hemiptera: Cicadellidae), two research lines were addressed. The first one aimed to determine the indirect effects of maize infection by MBSP on some biological and behavioral parameters of the vector, whereas a second line investigated direct interactions of the phytoplasma with D. maidis during its movement through the vector body following acquisition from plants, and associated microbiota. Indirect effects were investigated in choice experiments in which alighting and oviposition preferences by D. maidis were compared on healthy vs. MBSP-infected plants with variable incubation time (diseased plants with early and advanced symptoms, or still asymptomatic). Likewise, indirect effect of MBSP on the D. maidis biology was determined in two life table experiments in which the vector was reared on healthy vs. MBSP-infected plants expressing advanced disease symptoms or still asymptomatic. Choice experiments showed that alighting and oviposition preferences of D. maidis on MBSP-infected plants compared to healthy plants depend on the pathogen incubation period in the plant. The leafhopper preferred MBSP-infected plants over healthy ones during the asymptomatic phase of the disease, but rejected infected plants with advanced symptoms. The vector was able to acquire MBSP from asymptomatic infected plants shortly (3 days) after inoculation, but transmission efficiency increased when acquisition occurred at later stages of the pathogen incubation period (≥14 days) in the source plants and the test plants showed disease symptoms faster. These results suggest that MBSP modulates D. maidis preference for asymptomatic infected plants in the early stages of the crop, allowing rapid spread of this pathogen. Maize infection by the phytoplasma had a neutral effect on most life table parameters of D. maidis; a lower net reproductivity rate (Ro) was observed in the cohort reared on MBSP-infected plants with advanced symptoms, which was compensated to some extent by a higher sexual ratio. MBSP acquisition by all vector nymphal stadia was confirmed by PCR, and the pathogen as detected in both male and female reproductive organs. Concerning direct MBSP-vector interactions, transmission electron microscopy analyses showed phytoplasma-like cells in the midgut lumen, microvilli and epithelial cells, suggesting that MBSP enters the epithelium midgut through the microvilli wall. Within the epithelial cells, mitochondria and bacteria-like cells (possibly endosymbionts) were observed together with masses of phythoplasma-like cells. In the hemocoel, phytoplasma-like cells grouped into a matrix were also observed in association with bacteria-like cells similar to those observed in the midgut epithelium. Similar associations were found in the salivary gland. Interestingly, in-situ hybridization (FISH) technique revealed a variation in diversity and abundance of the microbiota in intestine and salivary glands of D. maidis adults over time after MBSP acquisition from plants. Sulcia sp., Cardinium sp. and eubacteria increased their abundance over time, whereas Rickettsia sp. decreased. The frequent association of the vector microbiota with the phytoplasma in some tissues of D. maidis suggests that endosymbiotic bacteria may play some role in MBSP-vector interactions.

Hepatitis B surface antigen vector delivers protective cytotoxic T-lymphocyte responses to disease-relevant foreign epitopes

Although hepatitis B surface antigen (HBsAg) per se is highly immunogenic, its use as a vector for the delivery of foreign cytotoxic T-lymphocyte (CTL) epitopes has met with little success because of constraints on HBsAg stability and secretion imposed by the insertion of foreign sequence into critical hydrophobic/amphipathic regions. Using a strategy entailing deletion of DNA encoding HBsAg-specific CTL epitopes and replacement with DNA encoding foreign CTL epitopes, we have derived chimeric HBsAg DNA immunogens which elicited effector and memory CTL responses in vitro, and pathogen- and tumor-protective responses in vivo, when the chimeric HBsAg DNAs were used to immunize mice. We further show that HBsAg DNA recombinant for both respiratory syncytial virus and human papillomavirus CTL epitopes elicited simultaneous responses to both pathogens. These data demonstrate the efficacy of HBsAg DNA as a vector for the delivery of disease-relevant protective CTL responses. They also suggest the applicability of the approach of deriving chimeric HBsAg DNA immunogens simultaneously encoding protective CTL epitopes for multiple diseases. The DNAs we tested formed chimeric HBsAg virus-like particles (VLPs). Thus, our results have implications for the development of vaccination strategies using either chimeric HBsAg DNA or VLP vaccines. HBsAg is the globally administered vaccine for hepatitis B virus infection, inviting its usage as a vector for the delivery of immunogens from other diseases.

On Root Arrangements of Polynomial-Like Functions and their Derivatives

2000 Mathematics Subject Classification: 12D10.

Design and validation of a novel generic platform for the production of tetravalent IgG1-like bispecific antibodies

International audience

Quarkonia and heavy-light mesons in a covariant quark model

Preliminary calculations using the Covariant Spectator Theory (CST) employed a scalar linear confining interaction and an additional constant vector potential to compute the mesonic mass spectra. In this work we generalize the confining interaction to include more general structures, in particular a vector and also a pseudoscalar part, as suggested by a recent study. A one-gluon-exchange kernel is also implemented to describe the short-range part of the interaction. We solve the simplest CST approxima- tion to the complete Bethe-Salpeter equation, the one-channel spectator equation, using a numerical technique that eliminates all singularities from the kernel. The parameters of the model are determined through a fit to the experimental pseudoscalar meson spectra, with a good agreement for both quarkonia and heavy-light states.

Quark model with chiral-symmetry breaking and confinement in the Covariant Spectator Theory

We propose a model for the quark-antiquark interaction in Minkowski space using the Covariant Spectator Theory. We show that with an equal-weighted scalar-pseudoscalar structure for the confining part of our interaction kernel the axial-vector Ward-Takahashi identity is preserved and our model complies with the Adler-zero constraint for π-π-scattering imposed by chiral symmetry.