997 resultados para degenerate primers
Resumo:
Estudi realitzat a partir d’una estada al Physics Department de la New York University, United States, Estats Units, entre 2006 i 2008. Una de les observacions de més impacte en la cosmologia moderna ha estat la determinació empírica que l’Univers es troba actualment en una fase d’Expansió Accelerada (EA). Aquest fenòmen implica que o bé l’Univers està dominat per un nou sector de matèria/energia, o bé la Relativitat General deixa de tenir validesa a escales cosmològiques. La primera possibilitat comprèn els models d’Energia Fosca (EF), i el seu principal problema és que l’EF ha de tenir propietats tan especials que es fan difícils de justificar teòricament. La segona possibilitat requereix la construcció de teories consistents de Gravetat Modificada a Grans Distàncies (GMGD), que són una generalització dels models de gravetat massiva. L’interès fenomenològic per aquestes teories també va resorgir amb l’aparició dels primers exemples de models de GMGD, com ara el model de Dvali, Gabadadze i Porrati (DGP), que consisteix en un tipus de brana en una dimensió extra. Malauradament, però, aquest model no permet explicar de forma consistent l’EA de l’Univers. Un dels objectius d’aquest projecte ha estat establir la viabilitat interna i fenomenològica dels models de GMGD. Des del punt de vista fenomenològic, ens hem centrat en la questió més important a la pràctica: trobar signatures observacionals que permetin distingir els models de GMGD dels d’EF. A nivell més teòric, també hem investigat el significat de les inestabilitats del model DGP.L’altre gran objectiu que ens vam proposar va ser la construcció de noves teories de GMGD. En la segona part d’aquest projecte, hem elaborat i mostrat la consistència del model “DGP en Cascada”, que generalitza el model DGP a més dimensions extra, i representa el segon model consistent i invariant-Lorentz a l’espai pla conegut. L’existència d’altres models de GMGD més enllà de DGP és de gran interès atès que podria permetre obtenir l’EA de l’Univers de forma purament geomètrica.
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Des de que començà aquest projecte, el grup de recerca ha intentat aprofundir el coneixement de la Catalunya de la Guerra del Francès (1808 -1814) a partir d’una òptica britànica. El grup pretenia així desenvolupar la relació que es va establir entre els catalans i els britànics al llarg de tota la guerra, des dels primers contactes permesos per la presència de la flota britànica en la costa catalana fins a la intervenció de forces britàniques en territori català. D’aquesta manera, i primerament, el grup inicià la consulta de les bases de dades i catàlegs catalans i britànics per a completar el nostre llistat de referències arxivístiques i bibliogràfiques. El segon pas van ésser les tres estades d’investigació que entre el 2006 i el 2007 es van fer a Anglaterra, principalment a Londres. La investigació es realitzà a la British Library, al Institute of Historical Research of the School of Advanced Studies de la University of London, al National Maritime Museum i als National Archives of the United Kingdom. A continuació, el grup analitzà la informació recollida de la lectura de fonts primàries i bibliogràfiques en aquests centres de recerca. Finalment, el grup creu que la intensa relació que es va establir entre les dues parts, reflecteix la importància que les autoritats britàniques van donar a Catalunya, i que el seu aïllament amb el centre polític del bàndol patriota va permetre que desenvolupés les seves pròpies dinàmiques i cronologies, encara que s’integraven en el desenvolupament general de la guerra.
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This paper investigates dynamic completeness of financial markets in which the underlying risk process is a multi-dimensional Brownian motion and the risky securities dividends geometric Brownian motions. A sufficient condition, that the instantaneous dispersion matrix of the relative dividends is non-degenerate, was established recently in the literature for single-commodity, pure-exchange economies with many heterogenous agents, under the assumption that the intermediate flows of all dividends, utilities, and endowments are analytic functions. For the current setting, a different mathematical argument in which analyticity is not needed shows that a slightly weaker condition suffices for general pricing kernels. That is, dynamic completeness obtains irrespectively of preferences, endowments, and other structural elements (such as whether or not the budget constraints include only pure exchange, whether or not the time horizon is finite with lump-sum dividends available on the terminal date, etc.)
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Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics.
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A recent study with 69 Japanese liver transplants treated with tacrolimus found that the MDR13435 C >T polymorphism, but not the MDR12677 G >T polymorphism, was associated with differences in the intestinal expression level of CYP3A4 mRNA. In the present study, over 6 h, we measured the kinetics of a 75 microg oral dose of midazolam, a CYP3A substrate, in 21 healthy subjects genotyped for the MDR13435 C >T and 2677 G >T polymorphism. No statistically significant differences were found in the calculated pharmacokinetic parameters between the three 3435 C >T genotypes (TT, CT and CC group, respectively: Cmax (mean +/- SD: 0.30 +/- 0.08 ng/ml, 0.31 +/- 0.09 ng/ml and 0.31 +/- 0.11 ng/ml; Apparent clearance: 122 +/- 29 l/h, 156 +/- 92 l/h and 111 +/- 35 l/h; t1/2: 1.9 +/- 1.1 h, 1.6 +/- 0.90 h and 1.7 +/- 0.7 h). In addition, the 30-min 1'OH midazolam to midazolam ratio, a marker of CYP3A activity, determined in 74 HIV-positive patients before the introduction of antiretroviral treatment, was not significantly different between the three 3435 C >T genotypes (mean ratio +/- SD: 3.65 +/- 2.24, 4.22 +/- 3.49 and 4.24 +/- 2.03, in the TT, CT and CC groups, respectively). Similarly, no association was found between the MDR12677 G >T polymorphism and CYP3A activity in the healthy subjects or in the HIV-positive patients. The existence of a strong association between the activity of CYP3A and MDR13435 C >T and 2677 G >T polymorphisms appears unlikely, at least in Caucasian populations and/or in the absence of specific environmental factors.
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El procés de creació de empreses en Espanya es un procediment complex, que requereix un estudi en profunditat del projecte y una planificació. La decisió de crear una empresa, ha de partir de una idea de negoci innovadora, que satisfaci les necessitats dels clients. L’emprenedor deurà analitzar el pla de negoci y la seva viabilitat en un mercat competitiu per determinar si el projecte es suficientment sòlid com per portar-se a cap. Una vegada avaluat el funcionament de la empresa, l’empresari deu prendre la decisió de la forma de constitució de la entitat més adequada, en funció de les característiques i necessitats del projecte. Cada forma jurídica requereix una tramitació específica adequada a les característiques de la mateixa. El nombre de promotors, la responsabilitat dels emprenedors, la fiscalitat que aplica i el import mínim de capital social a aportar són les principals variables per la elecció de la forma jurídica. Abans del inici de la activitat, tota nova empresa ha de realitzar el tràmits que li corresponguin i complir amb les obligacions contables i fiscals estipulades. Aquesta tramitació s’agrupa en les següents categories: laborables, fiscals, llibre, registre, adopció de la forma jurídica y llicencies municipals. El cost y temps necessari per la realització de aquest procés, varia respecte la forma jurídica i les propietats. En la actualitat, ens troben amb un procediment extens i feixuc pels empresaris, que rellenteix i encareix el inici de la activitat. El dinamisme empresarial està estretament relacional amb la regulació, tant en tramitació com en fiscalitat, per tant, una metodologia carregada i complexa redueix la motivació e indueix al fracàs. Davant la situació financera i econòmica actual, una manera de incentivar als empresaris i fomentar el creixement socioeconòmic, es incorporant instruments que agilitzin i redueixin el cost de tramitació. La nova llei de societats limitades han suposat un avanç important respecte la tramitació de les noves societats. Mitjançant la incorporació dels punts de assessorament e inici a la tramitació (PAIT) i la tramitació via telemàtica, el empresari pot realitzar les gestions necessàries de la societat limitada de una manera àgil i menys costosa, i alhora, obtenir assessorament durant el procediment i els primers anys després de iniciar la activitat. En definitiva, el procés de creació de la nova empresa ha millorat, no obstant, convindrien certes modificacions en els procediments, a través de una política de simplificació administrativa, afavorint la fiscalitat de les noves empreses i facilitat el accés al finançament per a totes les formes jurídiques.
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Aquest projecte intenta proporcionar les bases per la selecció i implantació d’un ERP a una empresa mitjana. S’analitzen des dels primers passos a realitzar per l’empresa que adquirirà l’ERP fins els que realitza el proveïdor per garantir l’èxit de la implantació. Primerament, i des del punt de vista del client, es realitza una anàlisi interna per detectar la necessitat d’un canvi de sistema i es detalla una metodologia a seguir per a l’elecció de l’ERP i del proveïdor d’aquest, minimitzant el risc d’error. Des del punt de vista del proveïdor es seguirà una metodologia d’implantació pas per pas, ja què un canvi de sistema és una tasca molt complexa. He intentat aportar la meva experiència al realitzar aquest projecte ja que he tingut l'oportunitat de viure una implantació des de les dues òptiques i he aprofitat per indagar en les tasques que s’hi realitzen.
Resumo:
Treball de recerca realitzat per un alumne d'ensenyament secundari i guardonat amb un Premi CIRIT per fomentar l'esperit científic del Jovent l'any 2009. El projecte Meteotek08, s'inicia durant el mes de febrer del curs 07/08 i es deu a tres fets: la passió per l'electrònica i la informàtica, per la meteorologia i per l'espai. Així va sorgir la idea de la possibilitat de realitzar algun projecte que relacionés aquestes idees: una sonda meteorològica, capaç d'assolir altituds de més de 30.000m, fer un registre de les condicions atmosfèriques i, fins i tot, captar fotografies. Era una idea complexa de dur a terme, ja que es partia del res, excepte de l'experiència d'algun dels integrants en el camp de l'electrònica i la informàtica. Durant els primers mesos es van començar a crear els primers prototips i a aplicar les primeres idees, fins que malauradament el crèdit es va acabar el juny. Tot i això, el treball va continuar durant l'estiu, durant el qual es va deixar el projecte gairebé enllestit. Diversos problemes amb el mòdul de comunicacions via ràdio van comportar l'endarreriment del projecte fins el passat febrer de 2009. El llançament es va dur a terme el dia 28 de febrer. La població escollida va ser Bujaraloz, situada als Monegros, degut a la seva bona posició geogràfica. El llançament es va dur a terme amb èxit i el seguiment i recepció de dades es va fer amb 2 cotxes. Al cap de 2 hores i 10 minuts la sonda tocava terra de nou, i poca estona després era localitzada. Els resultats van ser totalment satisfactoris, i es van obtenir també bones fotografies.
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MHC class II (MHCII) molecules play a pivotal role in the induction and regulation of immune responses. The transcriptional coactivator class II transactivator (CIITA) controls MHCII expression. The CIITA gene is regulated by three independent promoters (pI, pIII, pIV). We have generated pIV knockout mice. These mice exhibit selective abrogation of interferon (IFN)-gamma-induced MHCII expression on a wide variety of non-bone marrow-derived cells, including endothelia, epithelia, astrocytes, and fibroblasts. Constitutive MHCII expression on cortical thymic epithelial cells, and thus positive selection of CD4(+) T cells, is also abolished. In contrast, constitutive and inducible MHCII expression is unaffected on professional antigen-presenting cells, including B cells, dendritic cells, and IFN-gamma-activated cells of the macrophage lineage. pIV(-/-) mice have thus allowed precise definition of CIITA pIV usage in vivo. Moreover, they represent a unique animal model for studying the significance and contribution of MHCII-mediated antigen presentation by nonprofessional antigen-presenting cells in health and disease.
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In cognition, common factors play a crucial role. For example, different types of intelligence are highly correlated, pointing to a common factor, which is often called g. One might expect that a similar common factor would also exist for vision. Surprisingly, no one in the field has addressed this issue. Here, we provide the first evidence that there is no common factor for vision. We tested 40 healthy students' performance in six basic visual paradigms: visual acuity, vernier discrimination, two visual backward masking paradigms, Gabor detection, and bisection discrimination. One might expect that performance levels on these tasks would be highly correlated because some individuals generally have better vision than others due to superior optics, better retinal or cortical processing, or enriched visual experience. However, only four out of 15 correlations were significant, two of which were nontrivial. These results cannot be explained by high intraobserver variability or ceiling effects because test-retest reliability was high and the variance in our student population is commensurate with that from other studies with well-sighted populations. Using a variety of tests (e.g., principal components analysis, Bayes theorem, test-retest reliability), we show the robustness of our null results. We suggest that neuroplasticity operates during everyday experience to generate marked individual differences. Our results apply only to the normally sighted population (i.e., restricted range sampling). For the entire population, including those with degenerate vision, we expect different results.
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We demonstrate that the cccB gene, identified in the Bacillus subtilis genome sequence project, is the structural gene for a 10-kDa membrane-bound cytochrome c(551) lipoprotein described for the first time in B. subtilis. Apparently, CccB corresponds to cytochrome c(551) of the thermophilic bacterium Bacillus PS3. The heme domain of B. subtilis cytochrome c(551) is very similar to that of cytochrome c(550), a protein encoded by the cccA gene and anchored to the membrane by a single transmembrane polypeptide segment. Thus, B. subtilis contains two small, very similar, c-type cytochromes with different types of membrane anchors. The cccB gene is cotranscribed with the yvjA gene, and transcription is repressed by glucose. Mutants deleted for cccB or yvjA-cccB show no apparent growth, sporulation, or germination defect. YvjA is not required for the synthesis of cytochrome c(551), and its function remains unknown.
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The development of additional methods for detecting and identifuing Babesia and Plasmodium infections may be useful in disease monitoring, management and control efforts. To preliminarily evaluate sunthetic peptide-based serodiagnosis, a hydrophilic sequence (DDESEFDKEK)was selected from published BabR gene of B. bovis. Immunization of rabbits and cattle with the hemocyanin-conjugated peptide elicited antibody responses that specifically detected both P. falciparum and B. bovis antigens by immunofluorescence and Western blots. Using a dot-ELISA with this peptide, antisera from immunized and naturally-infected cattle, and immunized rodents, were specifically detected. Reactivity was weak and correlated with peptide immunization or infection. DNA-based detection using repetitive DNA was species-specific in dot-blot formats for B. bovis DNA, and in both dot-blot and in situ formats for P. falciparum; a streamlined enzymelinked synthetic DNA assay for P. falciparum detected 30 parasites/mm(cúbicos) from patient blood using either colorimetric (2-15 h color development) or chemiluminescent detection (0.5-6-min. exposures). Serodiagnostic and DNA hybridization methods may be complementary in the respective detection of both chronic and acute infections. However, recent improvements in the polymerase chain reaction (PCR) make feasible a more sensitive and uniform approach to the diagnosis of these and other infectious disease complexes, with appropriate primers and processing methods. An analysis of ribosomal DNA genes of Plasmodium and Toxoplasma identified Apicomplexa-conserved sequence regions. Specific and distinctive PCR profiles were obtained for primers spanning the internal transcribed spacer locus for each of several Plasmodium and Babesia species.
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The present study was performed to assess the interlaboratory reproducibility of the molecular detection and identification of species of Zygomycetes from formalin-fixed paraffin-embedded kidney and brain tissues obtained from experimentally infected mice. Animals were infected with one of five species (Rhizopus oryzae, Rhizopus microsporus, Lichtheimia corymbifera, Rhizomucor pusillus, and Mucor circinelloides). Samples with 1, 10, or 30 slide cuts of the tissues were prepared from each paraffin block, the sample identities were blinded for analysis, and the samples were mailed to each of seven laboratories for the assessment of sensitivity. A protocol describing the extraction method and the PCR amplification procedure was provided. The internal transcribed spacer 1 (ITS1) region was amplified by PCR with the fungal universal primers ITS1 and ITS2 and sequenced. As negative results were obtained for 93% of the tissue specimens infected by M. circinelloides, the data for this species were excluded from the analysis. Positive PCR results were obtained for 93% (52/56), 89% (50/56), and 27% (15/56) of the samples with 30, 10, and 1 slide cuts, respectively. There were minor differences, depending on the organ tissue, fungal species, and laboratory. Correct species identification was possible for 100% (30 cuts), 98% (10 cuts), and 93% (1 cut) of the cases. With the protocol used in the present study, the interlaboratory reproducibility of ITS sequencing for the identification of major Zygomycetes species from formalin-fixed paraffin-embedded tissues can reach 100%, when enough material is available.
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BACKGROUND: Superinfection with drug resistant HIV strains could potentially contribute to compromised therapy in patients initially infected with drug-sensitive virus and receiving antiretroviral therapy. To investigate the importance of this potential route to drug resistance, we developed a bioinformatics pipeline to detect superinfection from routinely collected genotyping data, and assessed whether superinfection contributed to increased drug resistance in a large European cohort of viremic, drug treated patients. METHODS: We used sequence data from routine genotypic tests spanning the protease and partial reverse transcriptase regions in the Virolab and EuResist databases that collated data from five European countries. Superinfection was indicated when sequences of a patient failed to cluster together in phylogenetic trees constructed with selected sets of control sequences. A subset of the indicated cases was validated by re-sequencing pol and env regions from the original samples. RESULTS: 4425 patients had at least two sequences in the database, with a total of 13816 distinct sequence entries (of which 86% belonged to subtype B). We identified 107 patients with phylogenetic evidence for superinfection. In 14 of these cases, we analyzed newly amplified sequences from the original samples for validation purposes: only 2 cases were verified as superinfections in the repeated analyses, the other 12 cases turned out to involve sample or sequence misidentification. Resistance to drugs used at the time of strain replacement did not change in these two patients. A third case could not be validated by re-sequencing, but was supported as superinfection by an intermediate sequence with high degenerate base pair count within the time frame of strain switching. Drug resistance increased in this single patient. CONCLUSIONS: Routine genotyping data are informative for the detection of HIV superinfection; however, most cases of non-monophyletic clustering in patient phylogenies arise from sample or sequence mix-up rather than from superinfection, which emphasizes the importance of validation. Non-transient superinfection was rare in our mainly treatment experienced cohort, and we found a single case of possible transmitted drug resistance by this route. We therefore conclude that in our large cohort, superinfection with drug resistant HIV did not compromise the efficiency of antiretroviral treatment.
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Liver fatty-acid-binding protein (L-FABP) is a cytoplasmic polypeptide that binds with strong affinity especially to long-chain fatty acids (LCFAs). It is highly expressed in both the liver and small intestine, where it is thought to have an essential role in the control of the cellular fatty acid (FA) flux. Because expression of the gene encoding L-FABP is increased by both fibrate hypolipidaemic drugs and LCFAs, it seems to be under the control of transcription factors, termed peroxisome-proliferator-activated receptors (PPARs), activated by fibrate or FAs. However, the precise molecular mechanism by which these regulations take place remain to be fully substantiated. Using transfection assays, we found that the different PPAR subtypes (alpha, gamma and delta) are able to mediate the up-regulation by FAs of the gene encoding L-FABP in vitro. Through analysis of LCFA- and fibrate-mediated effects on L-FABP mRNA levels in wild-type and PPARalpha-null mice, we have found that PPARalpha in the intestine does not constitute a dominant regulator of L-FABP gene expression, in contrast with what is known in the liver. Only the PPARdelta/alpha agonist GW2433 is able to up-regulate the gene encoding L-FABP in the intestine of PPARalpha-null mice. These findings demonstrate that PPARdelta can act as a fibrate/FA-activated receptor in tissues in which it is highly expressed and that L-FABP is a PPARdelta target gene in the small intestine. We propose that PPARdelta contributes to metabolic adaptation of the small intestine to changes in the lipid content of the diet.