José Manuel Guisán

Modulation of the enantioselectivity of lipases via controlled immobilization and medium engineering: hydrolytic resolution of mandelic acid esters

Abstract Lipase from Candida rugosa (CRL) has been purified and immobilized by using different immobilization protocols: interfacial adsorption on hydrophobic supports, ionic adsorption on PEI-coated supports, and covalent immobilization (on glutaraldehyde supports). This gave enzyme immobilized with different orientations and microenvironments. The catalytic properties (activity, specificity, and enantioselectivity) of the different derivatives have been found to be dramatically different. Very significant changes on activity with different substrates were found. For example, interfacially adsorbed derivative was the most active using simple substrates (ethyl butyrate) while PEI derivative was the most active hydrolysing ionic substrates (2-phenyl-2-butyroylacetic acid at pH 7) or methyl mandelate. The E value also depends strongly on the derivative and the conditions employed. Thus, the interfacially absorbed enzyme varied its enanatioselectivity (toward S isomer) from 1.6 to 85 in the hydrolysis of ( R , S )-2-phenyl-2-butyroylacetic acid when the pH value varied from 7 to 5. However, the glutaraldehyde derivative presented a high enantioselectivity ( E =400) toward R isomer (the inverse E value compared to the previous derivative) at both pH conditions. Polyethyleneimine (PEI) derivative presented a slight enantiopreference toward the S isomer. Thus, using different derivatives, it has been possible to obtain both pure enantiomers from the ester or the product. Similar changes in the E values were obtained in the hydrolysis of methyl mandelate though here always there was a enantiopreference for the S isomer. Using this substrate, the best derivative was the PEI derivative at pH 5 ( E =300), while the glutaraldehyde one presented an E value of only 10.

Synthesis of antibiotics (cephaloglycin) catalyzed by penicillin G acylase: Evaluation and optimization of different synthetic approaches

Abstract Two different approaches have been utilized to synthesize cephaloglycin using immobilized-stabilized penicillin G acylase derivatives. These are thermodynamically and kinetically controlled strategies. The thermodynamically controlled strategy could be employed to synthesize cephalotin or penicillin G, but this approach in the synthesis of cephaloglycin presented serious difficulties because of the absence of conditions where the thermodynamics of the process and the enzyme activity/stability properties were good enough. The kinetically controlled strategy has given much better results. The systematic study of the different parameters that defined the maximum yields of this strategy has permitted the identification of its main problem as the hydrolysis of the antibiotic. Because of the rapid enzymatic hydrolysis of cephaloglycin that has been previously synthetized, the yields were poor and they decreased very rapidly after reaching the maximum yield. Three different strategies have been used to decrease this amidase activity (an excess of acyl donor, selection of acidic pH, and distortion of the enzyme molecule by methanol). Simultaneous utilization of these strategies has significantly improved this synthetic process with very high yields (around 95%), reaction rates, and enzyme stability.

Selective oxidation: stabilisation by multipoint attachment of ferredoxin NADP+ reductase, an interesting cofactor recycling enzyme

Ferredoxin-NADP+ reductase (FNR, EC 1.18.1.2) is an enzyme that is able to catalyse the oxidation of NADPH + H+. A strategy to prepare industrial derivatives of this enzyme for use as an ‘NADP’ regenerating enzyme in oxidizing reactions is presented. The strategy is based on a strictly controlled process of multipoint covalent attachment between the enzyme, via its amino groups, and a pre-existing solid activated with a monolayer of simple aldehyde groups linked by a space-arm of moderate length to the surface of the support. Controlling the variables which may have an influence in the multi-interaction process, we have prepared a number of enzyme derivatives with very different activity/stability properties. The ‘optimum derivative’ was found to be much more stable than its corresponding soluble enzyme under all the denaturation conditions assayed (high temperatures, extreme pH, organic solvents, etc.). Because of the excellent properties of this enzyme derivative, we can regenerate NADP+ by using molecular oxygen directly as the oxidizing agent under a wide range of conditions. Coupling this oxidative system to other NADP-dependent redox enzymes, we should be able to develop a very specific and selective oxidative procedure under very mild oxidizing conditions.

Oriented covalent immobilization of antibodies onto heterofunctional agarose supports: a highly efficient immuno-affinity chromatography platform.

The development of new bioconjugates formed by one antibody optimally bound (through its Fc region) to fairly inert solid surfaces is of primary relevance in immuno-affinity chromatography. Immunoglobulins G (IgG) have a Fc region very rich in histidine (His) residues. In this way, immobilization of IgGs on heterofunctional metal chelate-glyoxyl supports (Ag-Me(2+)/G) takes place in two steps: firstly the antibodies are conjugated to the support via His-metal coordination bonds. Secondly, their incubation under alkaline condition promotes an intramolecular covalent attachment between lysine residues at the Fc region and glyoxyl groups on the support surface. The IgG that recognizes as antigen the HRP (antiHRP-IgG) has been conjugated to Ag-Me(2+)/G supports. The resulting bioconjugate is highly inert and able to specifically bind the antigen (HRP) without significant unspecific binding of any other proteins, resulting in an excellent HRP purification platform. The binding activity of this bioconjugate has been optimized by controlling the antibody distribution throughout the beads surface in order to avoid high antibody densities that led to a low binding activity of the antibodies. The optimal antibody distribution has been achieved when these proteins were slowly immobilized on Ag-Cu(2+)/G in presence of imidazole. This bioconjugate was able to bind up to 1.5 moles of antigen per mole of antibody, only 1.3-fold less than the antibody in solution. Hence, we have been able to develop an optimal protocol to prepare bioconjugated composites in an oriented and irreversible fashion which results in highly efficient and specific surfaces for the exclusive biological recognition.

Enzymatic β-galactosidation of β-xylopyranosides

SummaryThe disaccharides formed by enzymatic transfer of the β-D-galactopyranosyl residue fromo-nitrophenyl β-d-galactopyranoside to β-d-xylopyranosides have been identified. The influence of different factors on the yields of the disaccharides obtained was evaluated. Significant changes in selectivity were observed when β-galactosidase fromE. coli was used instead of β-galactosidase fromA. oryzae.

Engineering of Enzymes via Immobili-zation and Post-Immobilization Techniques: Preparation of Enzyme Derivatives with Improved Stability in Organic Media

The high potential of enzyme biotransformations in nonaqueous media has been adequately emphasised throughout the present volume. However, enzymes also have important limitations for working in non-aqueous media. For example, enzymes are usually inactivated in the presence of high concentrations of organic cosolvents or in the presence of hydrophobic interfaces of non-miscible solvents and so on[1].Such limitations may not be very relevant when working at laboratory scale (e.g. by performing a short unique biotransformation) but they can become critical when trying to scale up such exciting biotransformations to an industrial scale (e.g. trying to perform a number of long reaction cycles)

Purification, Immobilization, Stabilization and Characterization of Commercial Extract with β-galactosidase Activity

From a commercial enzyme extract juice with purpose to stabilize it in the industry, has been able to extract an extract rich in beta-galactosidase activity. It has been designed to extract and purification protocol based on polyethyleneimine SUPPORTS agarose activated with next purification yields 80%. Subsequently it has been immobilized in different SUPPORTS, with yields of 95% in mobilization. To estabililzar dimensional structure of the enzyme in the supports by multipoint covalent immobilization, are subsequently incubated at pH 10. Both groups activated SUPPORTS glioxyl, has remained almost unchanged initial enzyme activity, whereas in the derivative BrCN activity was not observed after this process. Once completely inactivated derivatives, these have been thoroughly washed with water, and then placed in reaction initial conditions (pH, ionic strength) and all derivatives, have managed to recover more than 80% of the initial activity, and reused again for these new cycles reaction.

Diseño de nuevos catalizadores de amino oxidasas y catalasas co-inmovilizadas para la eliminación de aminas biógenas en vinos

The homeobox gene extradenticle (exd) acts as a cofactor of the homeotic genes in the specification of larval patterns during embryogenesis. To study its role in adult patterns, we have generated clones of mutant exd- cells and examined their effect on the different body parts. In some regions, exd- clones exhibit homeotic transformations similar to those produced by known homeotic mutations such as Ultrabithorax (Ubx), labial (lab), spineless-aristapedia (ssa) or Antennapedia (Antp). In other regions, the lack of exd causes novel homeotic transformations producing ectopic eyes and legs. Moreover, exd is also required for functions normally not associated with homeosis, such as the maintenance of the dorsoventral pattern, the specification of subpatterns in adult appendages or the arrangement of bristles in the mesonotum and genitalia. Our findings indicate that exd is critically involved in adult morphogenesis, not only in the homeotic function but also in several other developmental processes.Previously published experiments have shown that the endogenous Dfd gene can be ectopically activated by its own (heat-shock-driven) product in a subset of cells of different segments. This results in the differentiation of maxillary structures like cirri and mouth hooks in places where they normally do not appear, and represents a phenomenon of autocatalysis of homeotic gene function that differs from the normal activation process. We show that this out-of-context activation occurs in cells belonging to the anterior compartments of the three thoracic and the A1 to A8 abdominal segments and that it requires the normal function of the polarity genes wingless (wg) and engrailed (en). The wg product, in addition to that of Dfd, appears to be sufficient to activate the endogenous Dfd gene in many embryonic cells. We have studied the effect of several homeotic genes on Dfd activation and phenotypic expression: Scr, Antp, Ubx and Abd-B repress Dfd both transcriptionally and at the phenotypic level, if their products are in sufficient amounts. The endogenous abd-A gene does not have a noticeable effect, but when it is replaced by an hsp70-abd-A gene, which produces a high and uniform level of expression, the phenotypic expression of Dfd is suppressed. Our results also suggest that the differentiation of cirri is induced by Dfd-expressing cells in non-expressing neighboring cells, and that this interaction occurs across the parasegmental border.During evolution, many animal groups have developed specialised outgrowths of the body wall, limbs or appendages. The type of appendage depends on the identity of the segment where they appear, indicating that the Hox genes contribute to appendage specification. Moreover, work carried out principally in Drosophila has identified the gene products and the mechanisms involved in pattern formation in the appendages. In this essay, we compare the morphogenetic processes in the appendages and the body wall; the function of the Hox genes and the response to the signalling molecules involved in local patterning. We speculate that, although the basic mechanisms are similar, there are significant differences in the manner the body trunk and appendages respond to them.[ES] La pared celular es un elemento morfogenetico esencial que determina la forma final de las celulas y que las protege contra la lisis. En S. pombe esta esta constituida por ? y s-glucano y manoproteinas y tanto la sintesis como remodelacion de su estructura requiere de diferentes enzimas estrictamente reguladas. En S. pombe existe poca informacion de como se lleva a cabo la incorporacion del material de membrana y sobre la regulacion de las enzimas implicadas en la sintesis y remodelacion de la pared celular por los mecanismos de transporte vesicular. Para abordar el estudio de como el trafico vesicular mediado por clatrina afecta a la morfogenesis de S. pombe y en particular cual es su papel en la regulacion de la sintesis de la pared celular se ha analizado el papel tanto de la clatrina, mediante el analisis de diferentes mutantes de la cadena ligera de la clatrina, como el del adaptador AP-2, que interviene en el proceso de endocitosis mediada por clatrina. Se ha demostrado que la delecion de la cadena ligera de la clatrina resulta letal para las celulas de S. pombe y que esta letalidad se rescata al incubar las celulas en un medio suplementado con sorbitol. En este caso aunque las celulas pueden sobrevivir poseen graves defectos morfologicos, en crecimiento, en trafico vesicular, en desarrollo sexual, etc. Se ha podido comprobar que la ausencia de Clc1p afecta drasticamente a la estabilidad de Chc1p hecho que hace que, a diferencia de otros organismos, la supervivencia de S. pombe sea mas dependiente de la presencia clatrina. Ademas se ha demostrado que la letalidad causada por la ausencia de Clc1p se debe principalmente a defectos graves en la sintesis de la pared celular que afectan directamente a la sintesis del glucano. Los resultados obtenidos muestran que una reduccion en la cantidad de clatrina causa un leve impacto en el transporte vesicular en general y en otros procesos y elementos biologicos, pero afecta gravemente a la secrecion de enzimas de sintesis/remodelacion de la pared celular, como las s(1,3)glucan sintasa y endoglucanasas. En cuanto al complejo adaptador AP-2 se ha comprobado, que a diferencia de lo que se conoce hasta el momento en otros organismos unicelulares, este forma un complejo con la clatrina y se ha demostrado que tiene un papel en la endocitosis general de S. pombe. Asi mismo se ha descubierto que AP-2 puede estar interviniendo en la sintesis de la pared celular ya que su ausencia afecta a la actividad s-glucan sintasa y hace que S. pombe sea hiper-sensible a compuestos que afectan a la integridad de la pared celular.We characterized a novel protein of the Ras family, p19 (H-RasIDX). The c-H-ras proto-oncogene undergoes alternative splicing of the exon termed IDX. We show that the alternative p19 mRNA is stable and as abundant as p21 (p21 H-Ras4A) mRNA in all of the human tissues and cell lines tested. IDX is spliced into stable mRNA in different mammalian species, which present a high degree of nucleotide conservation. Both the endogenous and the transiently expressed p19 protein are detected in COS-1 and HeLa cells and show nuclear diffuse and speckled patterns as well as cytoplasmic localization. In yeast two-hybrid assays, p19 did not interact with two known p21 effectors, Raf1 and Rin1, but was shown to interact with RACK1, a scaffolding protein that promotes multiprotein complexes in different signaling pathways. This observation suggests that p19 and p21 play differential and complementary roles in the cell.Resumen del trabajo presentado al Congreso Nacional de Biotecnologia, celebrado en Murcia del 18 al 21 de junio de 2017.A. G. G. thanks Ramon Areces Foundation for a grant. J. C. thanks NIH-CA24487 for financial support.Ministerio de Educacion y Ciencia and grant S-0505/MAT-0283 from Comunidad Autonoma de Madrid to M.S. and by an Institutional grant from Fundacion Ramon Areces to the Centro de Biologia Molecular “Severo Ochoa”We report a genetic and molecular study of UbxMX6 and Ubx195rx1, two mutations in the Ultrabithorax (Ubx) locus which appear to have a strong effect on the activity of the homologous Ubx gene. These mutations show the characteristic embryonic and adult phenotypes of Ubx null alleles, and also fail to produce any detectable Ubx product. Yet, genetic and phenotypic analyses involving a large number of trans heterozygous combinations of UbxMX6 and Ubx195rx1 with different classes of Ubx mutations, indicate that they hyperactivate the homologous gene. This effect is induced on wildtype or mutant forms of Ubx, provided that the pairing in the bithorax region is normal, i.e. these mutations have a strong positive effect on transvection. We also show that, unlike all the other known cases of transvection in Ubx, this is not zeste-dependent. Southern analyses indicate that UbxMX6 is a 3.4 kb deletion, and Ubx195rx1 is an approximately 11 kb insertion of foreign DNA, both in the promoter region. We speculate that the region altered in the mutations may have a wildtype function to ensure cis-autonomy of the regulation of Ubx transcription.Resumen del trabajo presentado al Congreso Nacional de Biotecnologia, celebrado en Murcia del 18 al 21 de junio de 2017.The pannier (pnr) gene of Drosophila encodes a zinc-finger transcription factor of the GATA family and is involved in several developmental processes during embryonic and imaginal development. We report some novel aspects of the regulation and function of pnr during embryogenesis. Previous work has shown that pnr is activated by decapentaplegic (dpp) in early development, but we find that after stage 10, the roles are reversed and pnr becomes an upstream regulator of dpp. This function of pnr is necessary for the activation of the Dpp pathway in the epidermal cells implicated in dorsal closure and is not mediated by the JNK pathway, which is also necessary for Dpp activity in these cells. In addition, we show that pnr behaves as a selector-like gene in generating morphological diversity in the dorsoventral body axis. It is responsible for maintaining a subdivision of the dorsal half of the embryo into two distinct, dorsomedial and dorsolateral, regions, and also specifies the identity of the dorsomedial region. These results, together with prior work on its function in adults, suggest that pnr is a major factor in the genetic subdivision of the body of Drosophila.10th International Symposium on Reproductive Physiology of Fish (10th ISRPF), Expanding the khowledge base of reproductive success: from genes to the environment, 25-30 May 2014, Olhao, Portugal.-- 1 pageBy using a hsp70-Ubx fusion gene, we have ectopically expressed a Ubx product in the embryonic head primordia and studied the developmental effects on the larval head. We find that after high and persistent levels of Ubx product, the head is replaced by three (C1, C2 and C3) abdominal-like denticle belts. The C2 and C3 belts are the homeotic transformations of parasegments 1 and 2, respectively, while the C1 belt probably derives from the transformation and subsequent fusion of the most anterior procephalic primordia. On the basis of their response to the Ubx product and other arguments, we propose that the larval head is made of two genetically distinct components; one is the procephalon and the anterior region of the mandibular lobe, and the other is part of the parasegmental trunk and includes parasegments 1 and 2. Our results also indicate that most or all the larval head structures derive from precursor cells of ventral origin.The Iroquois (Iro) family of genes are found in nematodes, insects and vertebrates. They usually occur in one or two genomic clusters of three genes each and encode transcriptional controllers that possess a characteristic homeodomain. The Iro genes function early in development to specify the identity of diverse territories of the body, such as the dorsal head and dorsal mesothorax of Drosophila and the neural plate of Xenopus. In some aspects they act in the same way as classical selector genes, but they display specific properties that place them into a category of their own. Later in development in both Drosophila and vertebrates, the Iro genes function again to subdivide those territories into smaller domains.The pannier (pnr) gene encodes a GATA transcription factor and acts in several developmental processes in Drosophila, including embryonic dorsal closure, specification of cardiac cells and bristle determination. We show that pnr is expressed in the mediodorsal parts of thoracic and abdominal segments of embryos, larvae and adult flies. Its activity confers cells with specific adhesion properties that make them immiscible with non-expressing cells. Thus there are two genetic domains in the dorsal region of each segment: a medial (MED) region where pnr is expressed and a lateral (LAT) region where it is not. The homeobox gene iroquois (iro) is expressed in the LAT region. These regions are not formed by separate polyclones of cells, but are defined topographically. We show that ectopic pnr in the wing induces MED thoracic development, indicating that pnr specifies the identity of the MED regions. Correspondingly, when pnr is removed from clones of cells in the MED domain, they sort out and apparently adopt the LAT fate. We propose that (1) the subdivision into MED and LAT regions is a general feature of the Drosophila body plan and (2) pnr is the principal gene responsible for this subdivision. We argue that pnr acts like a classical selector gene but differs in that its expression is not propagated through cell divisions.We have developed a specific polyclonal antibody that recognizes the protein products of the abdominal-A (abd-A) gene, a member of the bithorax complex of Drosophila. The normal expression domain extends from parasegments 7 to 13, in good correspondence with previous genetic and molecular results. However, while the anterior border of expression is precisely demarcated by a parasegmental boundary, the posterior border does not coincide with a lineage boundary. Within the normal domain, the expression of abd-A shows intrametameric modulation; the amount of product is higher in posterior compartments and in the most anterior cells of the anterior compartments and then gradually decreases. We have examined the effect on abd-A expression of a number of mutations, some mapping within and others outside the abd-A transcription unit. Those mapping to the transcription unit eliminate or severely reduce the amount of abd-A antigen, while those mapping outside produce an abnormal distribution of abd-A protein. Finally, we show that the abd-A gene is down-regulated in part of the Abdominal-B (Abd-B) domain, precisely in those regions where the Abd-B gene is expressed at high levels.Resumen del trabajo presentado al Yeast Genetics Meeting, celebrado en Stanford, California (USA) del 22 al 26 de agosto de 2018.The effect of the anti-tumoral drug lauryl gallate on the infectivity of the African swine fever virus among other DNA (Herpes simplex and Vaccinia) and RNA (Influenza, Porcine transmissible gastroenteritis and Sindbis) viruses, involved in animal and human diseases, is analyzed. Viral production was strongly inhibited in different cell lines at non-toxic concentrations of the drug (1-10 μM), reducing the titres from 3 to more than 5 log. units depending on the multiplicity of infection. In our model system (African swine fever virus in Vero cells), the addition of the drug 1 h before virus adsorption, completely abolished virus productivity in a one-step growth virus cycle. Interestingly, no inhibitory effect was observed when lauryl gallate was added after 5 to 8 hpi. Both cellular and viral DNA synthesis and late viral transcription were inhibited by the drug, but, however, the early viral protein synthesis and the virus-mediated increasing of p53 remained unaffected. Activation of the apoptotic effector caspase-3 was not detected after lauryl gallate treatment of Vero cells, and, furthermore, the presence of the drug abrogated the activation of this protease induced by the virus infection. The overall results likely indicate that a cellular factor/function might be the target of the antiviral action of alkyl gallates.Tesis Doctoral presentada por Eduardo Rodenas Martinez en el Centro Andaluz de Biologia del Desarrollo, centro mixto CSIC-UPO.Resumen del trabajo presentado al Yeast Genetics Meeting, celebrado en Stanford, California (USA) del 22 al 26 de agosto de 2018.

Inmovilización y estabilización de amino oxidasas para la eliminación de aminas biógenas en vino

The homeobox gene extradenticle (exd) acts as a cofactor of the homeotic genes in the specification of larval patterns during embryogenesis. To study its role in adult patterns, we have generated clones of mutant exd- cells and examined their effect on the different body parts. In some regions, exd- clones exhibit homeotic transformations similar to those produced by known homeotic mutations such as Ultrabithorax (Ubx), labial (lab), spineless-aristapedia (ssa) or Antennapedia (Antp). In other regions, the lack of exd causes novel homeotic transformations producing ectopic eyes and legs. Moreover, exd is also required for functions normally not associated with homeosis, such as the maintenance of the dorsoventral pattern, the specification of subpatterns in adult appendages or the arrangement of bristles in the mesonotum and genitalia. Our findings indicate that exd is critically involved in adult morphogenesis, not only in the homeotic function but also in several other developmental processes.Previously published experiments have shown that the endogenous Dfd gene can be ectopically activated by its own (heat-shock-driven) product in a subset of cells of different segments. This results in the differentiation of maxillary structures like cirri and mouth hooks in places where they normally do not appear, and represents a phenomenon of autocatalysis of homeotic gene function that differs from the normal activation process. We show that this out-of-context activation occurs in cells belonging to the anterior compartments of the three thoracic and the A1 to A8 abdominal segments and that it requires the normal function of the polarity genes wingless (wg) and engrailed (en). The wg product, in addition to that of Dfd, appears to be sufficient to activate the endogenous Dfd gene in many embryonic cells. We have studied the effect of several homeotic genes on Dfd activation and phenotypic expression: Scr, Antp, Ubx and Abd-B repress Dfd both transcriptionally and at the phenotypic level, if their products are in sufficient amounts. The endogenous abd-A gene does not have a noticeable effect, but when it is replaced by an hsp70-abd-A gene, which produces a high and uniform level of expression, the phenotypic expression of Dfd is suppressed. Our results also suggest that the differentiation of cirri is induced by Dfd-expressing cells in non-expressing neighboring cells, and that this interaction occurs across the parasegmental border.During evolution, many animal groups have developed specialised outgrowths of the body wall, limbs or appendages. The type of appendage depends on the identity of the segment where they appear, indicating that the Hox genes contribute to appendage specification. Moreover, work carried out principally in Drosophila has identified the gene products and the mechanisms involved in pattern formation in the appendages. In this essay, we compare the morphogenetic processes in the appendages and the body wall; the function of the Hox genes and the response to the signalling molecules involved in local patterning. We speculate that, although the basic mechanisms are similar, there are significant differences in the manner the body trunk and appendages respond to them.[ES] La pared celular es un elemento morfogenetico esencial que determina la forma final de las celulas y que las protege contra la lisis. En S. pombe esta esta constituida por ? y s-glucano y manoproteinas y tanto la sintesis como remodelacion de su estructura requiere de diferentes enzimas estrictamente reguladas. En S. pombe existe poca informacion de como se lleva a cabo la incorporacion del material de membrana y sobre la regulacion de las enzimas implicadas en la sintesis y remodelacion de la pared celular por los mecanismos de transporte vesicular. Para abordar el estudio de como el trafico vesicular mediado por clatrina afecta a la morfogenesis de S. pombe y en particular cual es su papel en la regulacion de la sintesis de la pared celular se ha analizado el papel tanto de la clatrina, mediante el analisis de diferentes mutantes de la cadena ligera de la clatrina, como el del adaptador AP-2, que interviene en el proceso de endocitosis mediada por clatrina. Se ha demostrado que la delecion de la cadena ligera de la clatrina resulta letal para las celulas de S. pombe y que esta letalidad se rescata al incubar las celulas en un medio suplementado con sorbitol. En este caso aunque las celulas pueden sobrevivir poseen graves defectos morfologicos, en crecimiento, en trafico vesicular, en desarrollo sexual, etc. Se ha podido comprobar que la ausencia de Clc1p afecta drasticamente a la estabilidad de Chc1p hecho que hace que, a diferencia de otros organismos, la supervivencia de S. pombe sea mas dependiente de la presencia clatrina. Ademas se ha demostrado que la letalidad causada por la ausencia de Clc1p se debe principalmente a defectos graves en la sintesis de la pared celular que afectan directamente a la sintesis del glucano. Los resultados obtenidos muestran que una reduccion en la cantidad de clatrina causa un leve impacto en el transporte vesicular en general y en otros procesos y elementos biologicos, pero afecta gravemente a la secrecion de enzimas de sintesis/remodelacion de la pared celular, como las s(1,3)glucan sintasa y endoglucanasas. En cuanto al complejo adaptador AP-2 se ha comprobado, que a diferencia de lo que se conoce hasta el momento en otros organismos unicelulares, este forma un complejo con la clatrina y se ha demostrado que tiene un papel en la endocitosis general de S. pombe. Asi mismo se ha descubierto que AP-2 puede estar interviniendo en la sintesis de la pared celular ya que su ausencia afecta a la actividad s-glucan sintasa y hace que S. pombe sea hiper-sensible a compuestos que afectan a la integridad de la pared celular.We characterized a novel protein of the Ras family, p19 (H-RasIDX). The c-H-ras proto-oncogene undergoes alternative splicing of the exon termed IDX. We show that the alternative p19 mRNA is stable and as abundant as p21 (p21 H-Ras4A) mRNA in all of the human tissues and cell lines tested. IDX is spliced into stable mRNA in different mammalian species, which present a high degree of nucleotide conservation. Both the endogenous and the transiently expressed p19 protein are detected in COS-1 and HeLa cells and show nuclear diffuse and speckled patterns as well as cytoplasmic localization. In yeast two-hybrid assays, p19 did not interact with two known p21 effectors, Raf1 and Rin1, but was shown to interact with RACK1, a scaffolding protein that promotes multiprotein complexes in different signaling pathways. This observation suggests that p19 and p21 play differential and complementary roles in the cell.Resumen del trabajo presentado al Congreso Nacional de Biotecnologia, celebrado en Murcia del 18 al 21 de junio de 2017.A. G. G. thanks Ramon Areces Foundation for a grant. J. C. thanks NIH-CA24487 for financial support.Ministerio de Educacion y Ciencia and grant S-0505/MAT-0283 from Comunidad Autonoma de Madrid to M.S. and by an Institutional grant from Fundacion Ramon Areces to the Centro de Biologia Molecular “Severo Ochoa”We report a genetic and molecular study of UbxMX6 and Ubx195rx1, two mutations in the Ultrabithorax (Ubx) locus which appear to have a strong effect on the activity of the homologous Ubx gene. These mutations show the characteristic embryonic and adult phenotypes of Ubx null alleles, and also fail to produce any detectable Ubx product. Yet, genetic and phenotypic analyses involving a large number of trans heterozygous combinations of UbxMX6 and Ubx195rx1 with different classes of Ubx mutations, indicate that they hyperactivate the homologous gene. This effect is induced on wildtype or mutant forms of Ubx, provided that the pairing in the bithorax region is normal, i.e. these mutations have a strong positive effect on transvection. We also show that, unlike all the other known cases of transvection in Ubx, this is not zeste-dependent. Southern analyses indicate that UbxMX6 is a 3.4 kb deletion, and Ubx195rx1 is an approximately 11 kb insertion of foreign DNA, both in the promoter region. We speculate that the region altered in the mutations may have a wildtype function to ensure cis-autonomy of the regulation of Ubx transcription.Resumen del trabajo presentado al Congreso Nacional de Biotecnologia, celebrado en Murcia del 18 al 21 de junio de 2017.The pannier (pnr) gene of Drosophila encodes a zinc-finger transcription factor of the GATA family and is involved in several developmental processes during embryonic and imaginal development. We report some novel aspects of the regulation and function of pnr during embryogenesis. Previous work has shown that pnr is activated by decapentaplegic (dpp) in early development, but we find that after stage 10, the roles are reversed and pnr becomes an upstream regulator of dpp. This function of pnr is necessary for the activation of the Dpp pathway in the epidermal cells implicated in dorsal closure and is not mediated by the JNK pathway, which is also necessary for Dpp activity in these cells. In addition, we show that pnr behaves as a selector-like gene in generating morphological diversity in the dorsoventral body axis. It is responsible for maintaining a subdivision of the dorsal half of the embryo into two distinct, dorsomedial and dorsolateral, regions, and also specifies the identity of the dorsomedial region. These results, together with prior work on its function in adults, suggest that pnr is a major factor in the genetic subdivision of the body of Drosophila.10th International Symposium on Reproductive Physiology of Fish (10th ISRPF), Expanding the khowledge base of reproductive success: from genes to the environment, 25-30 May 2014, Olhao, Portugal.-- 1 pageBy using a hsp70-Ubx fusion gene, we have ectopically expressed a Ubx product in the embryonic head primordia and studied the developmental effects on the larval head. We find that after high and persistent levels of Ubx product, the head is replaced by three (C1, C2 and C3) abdominal-like denticle belts. The C2 and C3 belts are the homeotic transformations of parasegments 1 and 2, respectively, while the C1 belt probably derives from the transformation and subsequent fusion of the most anterior procephalic primordia. On the basis of their response to the Ubx product and other arguments, we propose that the larval head is made of two genetically distinct components; one is the procephalon and the anterior region of the mandibular lobe, and the other is part of the parasegmental trunk and includes parasegments 1 and 2. Our results also indicate that most or all the larval head structures derive from precursor cells of ventral origin.The Iroquois (Iro) family of genes are found in nematodes, insects and vertebrates. They usually occur in one or two genomic clusters of three genes each and encode transcriptional controllers that possess a characteristic homeodomain. The Iro genes function early in development to specify the identity of diverse territories of the body, such as the dorsal head and dorsal mesothorax of Drosophila and the neural plate of Xenopus. In some aspects they act in the same way as classical selector genes, but they display specific properties that place them into a category of their own. Later in development in both Drosophila and vertebrates, the Iro genes function again to subdivide those territories into smaller domains.The pannier (pnr) gene encodes a GATA transcription factor and acts in several developmental processes in Drosophila, including embryonic dorsal closure, specification of cardiac cells and bristle determination. We show that pnr is expressed in the mediodorsal parts of thoracic and abdominal segments of embryos, larvae and adult flies. Its activity confers cells with specific adhesion properties that make them immiscible with non-expressing cells. Thus there are two genetic domains in the dorsal region of each segment: a medial (MED) region where pnr is expressed and a lateral (LAT) region where it is not. The homeobox gene iroquois (iro) is expressed in the LAT region. These regions are not formed by separate polyclones of cells, but are defined topographically. We show that ectopic pnr in the wing induces MED thoracic development, indicating that pnr specifies the identity of the MED regions. Correspondingly, when pnr is removed from clones of cells in the MED domain, they sort out and apparently adopt the LAT fate. We propose that (1) the subdivision into MED and LAT regions is a general feature of the Drosophila body plan and (2) pnr is the principal gene responsible for this subdivision. We argue that pnr acts like a classical selector gene but differs in that its expression is not propagated through cell divisions.We have developed a specific polyclonal antibody that recognizes the protein products of the abdominal-A (abd-A) gene, a member of the bithorax complex of Drosophila. The normal expression domain extends from parasegments 7 to 13, in good correspondence with previous genetic and molecular results. However, while the anterior border of expression is precisely demarcated by a parasegmental boundary, the posterior border does not coincide with a lineage boundary. Within the normal domain, the expression of abd-A shows intrametameric modulation; the amount of product is higher in posterior compartments and in the most anterior cells of the anterior compartments and then gradually decreases. We have examined the effect on abd-A expression of a number of mutations, some mapping within and others outside the abd-A transcription unit. Those mapping to the transcription unit eliminate or severely reduce the amount of abd-A antigen, while those mapping outside produce an abnormal distribution of abd-A protein. Finally, we show that the abd-A gene is down-regulated in part of the Abdominal-B (Abd-B) domain, precisely in those regions where the Abd-B gene is expressed at high levels.Resumen del trabajo presentado al Yeast Genetics Meeting, celebrado en Stanford, California (USA) del 22 al 26 de agosto de 2018.The effect of the anti-tumoral drug lauryl gallate on the infectivity of the African swine fever virus among other DNA (Herpes simplex and Vaccinia) and RNA (Influenza, Porcine transmissible gastroenteritis and Sindbis) viruses, involved in animal and human diseases, is analyzed. Viral production was strongly inhibited in different cell lines at non-toxic concentrations of the drug (1-10 μM), reducing the titres from 3 to more than 5 log. units depending on the multiplicity of infection. In our model system (African swine fever virus in Vero cells), the addition of the drug 1 h before virus adsorption, completely abolished virus productivity in a one-step growth virus cycle. Interestingly, no inhibitory effect was observed when lauryl gallate was added after 5 to 8 hpi. Both cellular and viral DNA synthesis and late viral transcription were inhibited by the drug, but, however, the early viral protein synthesis and the virus-mediated increasing of p53 remained unaffected. Activation of the apoptotic effector caspase-3 was not detected after lauryl gallate treatment of Vero cells, and, furthermore, the presence of the drug abrogated the activation of this protease induced by the virus infection. The overall results likely indicate that a cellular factor/function might be the target of the antiviral action of alkyl gallates.Tesis Doctoral presentada por Eduardo Rodenas Martinez en el Centro Andaluz de Biologia del Desarrollo, centro mixto CSIC-UPO.Resumen del trabajo presentado al Yeast Genetics Meeting, celebrado en Stanford, California (USA) del 22 al 26 de agosto de 2018.

Síntesis enzimática de lípidos estructurados (dicaprilil-monooleina). Papel crítico del biocatalizador

Tesis llevada a cabo para conseguir el grado de Doctor por la Universidad de Complutense de Madrid.--2018-01-15.--Sobresaliente cum laudeContiene 7 documentos (1. Objetivos, alcance y publicaciones. 2. Registro y codigo) y 5 con el softwareespanolLas ciber amenazas afectan a todo tipo de organizaciones, causando frecuentes y costosos impactos globalmente. Recientemente, han surgido productos de ciberseguro con el potencial de reducir el impacto de los riesgos en el ciberespacio. Sin embargo, aun tienen que madurar. En este articulo presentamos varios modelos de analisis de riesgos que podrian facilitar la implantacion y adopcion de ciberseguros. Estos modelos, descritos como diagramas de influencia y diagramas de influencia bi-agente, aportan un marco para estimar el impacto economico de los ciber riesgos a los que se enfrentan aseguradores y asegurados, asi como tambien para calcular sus estrategias optimas de mitigacion y transferencia del riesgo. EnglishCyber threats affect all kinds of organisations with frequent and costly impacts worldwide. Cyber insurance products have recently emerged with the potential of lowering the impact of cyberspace risks. However, they have yet to mature. In this paper we present several risk analysis models that may facilitate the implementation and adoption of cyber insurance. These models, described in terms of influence diagrams and bi-agent influence diagrams, provide a framework for estimating the economic impact of cyber risks that may face insurers and insurees as well as calculating their optimal risk mitigation and transfer strategies.Trabajo presentado a la III Iberoamerican Conference on Supercritical Fluids celebrada en Cartagena de Indias (Colombia) del 1 al 5 de abril de 2013.Trabajo presentado a la 26th IAEA Fusion Energy Conference, celebrada en Kyoto (Japon) del 17 al 22 de octubre de 2016.The Joint Iberian Meeting on Atomic and Molecular Physics (IBER), Barcelona, September 12-14th, 2017. -- http://iber2017.com/index.phpPoster presentado en el 10o Congreso de la Asociacion Iberica de Endocrinologia Comparada AIEC, celebrado en Castellon, Espana, del 23 al 25 de septiembre de 2015Resumen del trabajo presentado a la XI Reunion del Grupo de Microbiologia Molecular, celebrada en Sevilla del 6 al 8 de septiembre de 2016.Chinchilla-Rodriguez, Zaida; Miguel, Sandra; Perianes-Rodriguez, Antonio; Ovalle-Perandones, Antonia; Olmeda-Gomez, Carlos. (2016). Autonomy vs. dependency of scientific collaboration in scientific performance . 21st International Conference on Science and Technology Indicators, STI2016. Valencia, Spain, 14-16 September 2016XXV National Spectroscopy Meeting (XXV RNE), IX Iberian Spectroscopy Conference (IX CIE), Alicante, 20th-22nd July, 2016Resumen del trabajo presentado a la 43rd International Conference on Micro and Nano Engineering (MNE), celebrada en Braga (Portugal) del 18 al 22 de septiembre de 2017.Contiene 7 documentos (1. Objetivos, alcance y publicaciones. 2. Registro, interface y codigo) y 5 con el softwarePoster presentado en el congreso 4th SCARCE International Conference, celebrado en Cadiz, Espana el 25 y 26 de noviembre de 2013We present a Cultural Heritage conceptual data model built under the European INSPIRE ( Infrastructure for Spatial Information in the European Community ) Directive. Our model develops the Data Specification on Protected Sites , part of the INSPIRE Annex I. Hence its orientation towards georeferenced heritage data. The data model has been developed by an interdisciplinary group made up of specialists in the field of Geomatics and Cultural Heritage. It is our aim to achieve a generic, extendable and interoperable schema. It should be generic enough to embrace all kind of protected heritage data, from an ancient pilgrim’s way to the last artifact found in an archaeological excavation, extendable to allow any kind of data producer to adapt the model to the nature of their own information and interoperable to combine spatial data sets from different sources through network services, via Internet. The achievement of these three characteristics features international norms and standards referred to our kind of data. This implies adaptation to INSPIRE as well as to several ISO norms: ISO 19100 series regarding geographical information, ISO 21127 (CIDOC-CRM Model) for heritage thematic data and ISO 15836 (Dublin Core) for document resources. The data model comprehends two main dimensions: cultural entities in a strict sense, and the legislative figures created to protect them. This allows for the representation of cultural objects (i.e. historical buildings or archaeological sites) and their link to their legislative protection, keeping them as separate realities. In order to describe the data model, an international common language has been used: UML (Unified Modeling Language), a standard itself. Thus, we present a class-diagram depicting all legal and cultural entities, in the form of classes with their corresponding relations, attributes, constraints and stereotypes.Trabajo presentado en el 11th Spanish-Italian Symposium on Organic Chemistry SISOC-XI celebrado en San Sebastian (Espana), del 13 al 15 de julio de 2016.Trabajo presentado en el XVIII Congreso Peruano de Geologia, celebrado en Lima (Peru), del 16 al 19 de octubre de 2016Poster presentado en las XXXIII Jornadas de la Asociacion Espanola de Entomologia, celebradas en Almeria, del 15 al 18 de noviembre de 2017This research has been funded by the Spanish Ministry of Economy and Competitiveness, through research projects POSTFIRE (CGL2013-47862-C2-1-R) and GEOFIRE (CGL2012-38655-C04-01)The main contribution of this work is the improvement of the efficiency of a PEMFC power system while guaranteeing conditions that also improve its durability. Adopting the NMPC scheme with the distributed parameter model and the nonlinear observer, the efficiency of the PEMFC-based system can be maximized guaranteeing at the same time the appropriate internal gas concentration profiles to avoid global and local hydrogen and oxygen starvation and proper membrane humidification.Trabajo presentado en la 13a Reunion de la Red Espanola de Bacterias Lacticas (RedBAL), celebrada en Madrid (Espana) del 17 al 18 de junio de 2019Memoria de tesis doctoral presentada por Uxue Tilves Matheu para obtener el titulo de Doctora en Ciencias del Mar por la Universitat Politecnica de Catalunya (UPC), realizada bajo la direccion de la Dra. Ana Maria Sabates Freijo y de la Dra. Veronica Lorena Fuentes del Institut de Ciencies del Mar (ICM-CSIC).-- 149 pages, appendicesTrabajo presentado por Nuria Vallverdu Coll para la obtencion del grado de Master universitario en investigacion basica y aplicada en recursos cinegeticos, realizado en el Instituto de Investigacion en Recursos Cinegeticos (IREC, CSIC-UCLM-JCCM).Resumen del poster presentado a la 7th International Conference on Polyphenols and Health, celebrada en Tours (Francia) del 27 al 30 de octubre de 2015.Trabajo financiado con el Proyecto DR3AMCGL2014-55118 del MINECO (Proyecto coordinado del proyecto SALTACRES).Trabajo presentado en el Congreso Iberoamericano de Hidrogeno y Pilas de Combustible, celebrado en Barcelona (Espana) del 15 al 17 de octubre de 2014.El presente articulo describe el empleo del flujo de diseno basado en modelos para el desarrollo de bloques reconfigurables automaticamente para el procesado de imagenes sobre FPGA. Para ello se han concebido arquitecturas hardware que aprovechan caracteristicas especificas de algunos algoritmos de procesado y que pueden ser modificadas a traves de un novedoso procedimiento software. Este aspecto, unido a las restantes opciones de parametrizacion de los diferentes modulos, permite liberar al disenador de los detalles especificos de las implementaciones hardware asi como adaptar el consumo de recursos del FPGA a las necesidades de la aplicacion. El proceso de reconfiguracion automatica se ilustra con el bloque de convolucion generico realizando comparaciones entre implementaciones de diferentes arquitecturas sobre un FPGA Spartan-6 LX45.Trabajo presentado en el 8th European Meeting on Solar Chemistry and Photocatalysis, celebrado en Salonica (Grecia) del 25 al 28 de Junio de 2014.Resumen del poster presentado a las I Jornadas Cientificas del CIAL celebradas el 5 de junio de 2014 en Madrid.-- et al.Trabajo presentado en el 4th International Conodont Symposium, celebrado en Valencia (Espana), del 25 al 30 de junio de 2017Comunicacion presentada en el Aquaculture Europe 2014, celebrado en Donostia-San Sebastian, Espana, del 14 al 17 de octubre de 2014Triptico de la reunion celebrada en la Universidad de Murcia del 12 al 14 de Noviembre de 2015.Trabajo presentado en la 11a Reunion de la Red Espanola de Bacterias Lacticas (RedBAL), celebrada en Gijon (Asturias, Espana) del 28 al 30 de Junio de 2017Trabajo Fin de Master: Master en Catalisis Homogenea y Quimica Molecular. Grupo de Cristales liquidos y Polimeros, Departamento de Quimica Organica de la Universidad de Zaragoza (Curso 2014/2015).La elaboracion de este articulo se inscribe en el proyecto de investigacion “Innovacion oculta: cambio de paradigma en los estudios de innovacion” (FFI2011-25475), Ministerio de Economia y Competitividad, Gobierno de Espana.Poster presented at the 18th International Microscopy Congress (ICM 2014) that took place in Prague (Czech Republic) during 7-12th September 2014.Esta investigacion ha sido posible gracias a la financiacion de los proyectos del Plan Nacional CGL2012-36682 y CGL2016-75109-P del Ministerio de Economia y Competitividad, y al Convenio Principado de Asturias-Universidad de Oviedo y MNCN-CSIC.Resumen del poster presentado a la 10th International Conference one Carbon Metabolism, vitamins B and Homocysteine, celebrada en Nancy (Francia) del 7 al 11 de julio de 2015.La celulosa es la fuente de energia mas abundante que hay en la Tierra. Su transformacion en glucosa se considera la etapa mas importante en la produccion de biocombustibles a partir de biomasa lignocelulosica. Para esta transformacion es necesaria la accion sinergica de tres tipos de enzimas que hidrolizan los enlaces s-1,4 de la celulosa: (i) las endoglucanasas, que actuan al azar sobre enlaces internos, (ii) las celobiohidrolasas, que operan progresivamente por los extremos reductores y no reductores de la cadena, y (iii) las s-glucosidasas (BGL), que hidrolizan celobiosa y los celooligosacaridos mas pequenos hasta glucosa. Los hongos filamentosos son la principal fuente de celulasas, siendo los crudos del genero Trichoderma los mas estudiados y comercializados historicamente. Sin embargo, estos hongos secretan niveles insuficientes de BGL para una conversion efectiva de la celulosa, por lo que con frecuencia requieren ser suplementados con preparaciones ricas en esta enzima procedentes de otros hongos. Recientemente, se han descrito cepas de Penicillium con gran capacidad para secretar altos niveles de BGL al medio extracelular. Ademas de su papel en la hidrolisis de la celulosa las BGL tambien pueden emplearse para sintetizar compuestos de interes industrial mediante reacciones de transglicosilacion. En este trabajo se ha estudiado una nueva cepa fungica productora de celulasas, identificada como Talaromyces amestolkiae, en base a un analisis molecular y morfologico. Se han purificado a homogeneidad electroforetica tres BGL (BGL-1, BGL-2 y BGL-3) secretadas por el hongo y se han caracterizado bioquimicamente. BGL-1 y BGL-2 son proteinas monomericas, mientras que BGL-3 es un dimero funcional. Los valores de actividad maxima de estas enzimas se obtuvieron a pH 4,0 y entre 50-60 oC, siendo estables en un rango de pH de 4-7 y a 50 oC. Las tres mostraron distinto comportamiento en funcion del sustrato (pNPG o celobiosa), ensayandose tambien el efecto de determinados compuestos quimicos e inhibidores en su actividad. Las tres pueden hidrolizar celooligosacaridos de diferente longitud, disminuyendo la eficacia de hidrolisis con el aumento de la polimerizacion, y no son activas frente a polisacaridos. Ademas, mostraron actividad de transglicosilacion, formando alquilglicosidos y celooligosacaridos de mayor longitud que los usados como sustratos. Las tres BGL se identificaron mediante huella peptidica y en base la alta homologia con otras BGL fungicas relacionadas filogeneticamente, se disenaron cebadores especificos que permitieron la secuenciacion de los genes que codifican cada una de ellas. El analisis de las secuencias aminoacidicas mostro que BGL-1 es miembro de la familia 1 de las glicosil hidrolasas, mientras que BGL-2 y BGL-3 pertenecen a la familia 3. Teniendo en cuenta que existen pocas BGL fungicas cristalizadas, se construyeron modelos moleculares de estas ultimas en base a las estructuras con las que presentaron mayor identidad. El crudo enzimatico de T. amestolkiae, rico en BGL, se uso de forma individual y como suplemento de otros cocteles comerciales en experimentos de sacarificacion a partir de slurry acido de paja de trigo. Los resultados obtenidos indican que el hongo secreta, ademas de celulasas, otras enzimas complementarias que potencian la liberacion de azucares fermentables. Se analizo el secretoma de T. amestolkiae usando Avicel o slurry acido de paja de trigo como fuente de carbono. Las celulasas fueron las enzimas mas abundantes en ambas condiciones, sin embargo, en el crudo obtenido a partir de los cultivos en slurry aumento la proporcion de BGL y otras enzimas distintas a las celulasas, indicando que para la degradacion de un sustrato complejo es necesaria mayor diversidad enzimatica. Los resultados obtenidos en este trabajo abren nuevas vias para la formulacion de cocteles enzimaticos eficaces en el contexto de la degradacion de la biomasa lignocelulosica.Trabajo presentado en el 4th International Congress on Ichnology - ICHNIA 2016: Ichnology for the 21st century: (Palaeo) Biological Traces towards Sustainable Development, celebrado en Idanha-a-Nova (Portugal), del 6 al 9 de mayo de 2016Resumen del trabajo presentado al Joint European Magnetic Symposia (JEMS), celebrado en Glasgow (UK) del 21 al 26 de agosto de 2016.Vazquez, J. T. ... et. al.-- Inciativa Iberica para el Estudio de las Fallas Activas, Tercera Reunion Iberica sobre Fallas Activas y Paleosismologia IBERFAULT 2018 - Third Iberian Meeting on Active Faults and Paleoseismology, 11-13 June 2018, Alicante, Spain.-- 4 pages, 2 figuresLa infeccion temprana por Verticillium dahliae en olivar y la discriminacion entre niveles de severidad de la Verticilosis es viable mediante la utilizacion de imagenes termicas, multiespectrales e hiperespectrales adquiridas con vehiculos aereos no tripulados a escala de parcela o tripulados a escala de comarca. El indicador relacionado con la temperatura de copa (CWSI), indices de reflectancia (B, BG1, BR1) y la fluorescencia clorofilica (FLD3) se identifican como buenos indicadores para detectar la Verticilosis en etapas tempranas del desarrollo de la enfermedad, mientras que NDVI, PRI515, R/G, HI e indices de estimacion de clorofila y carotenos demuestran ser buenos indicadores para la cuantificacion de dano moderado o severo causada por Verticilosis, lo que puede ser de utilidad para el diseno de estrategias de control de la Verticilosis a escala de parcela y de comarca. La infeccion temprana por Verticillium dahliae en olivar y la discriminacion entre niveles de severidad de la Verticilosis es viable mediante la utilizacion de imagenes termicas, multiespectrales e hiperespectrales adquiridas con vehiculos aereos no tripulados a escala de parcela o tripulados a escala de comarca. El indicador relacionado con la temperatura de copa (CWSI), indices de reflectancia (B, BG1, BR1) y la fluorescencia clorofilica (FLD3) se identifican como buenos indicadores para detectar la Verticilosis en etapas tempranas del desarrollo de la enfermedad, mientras que NDVI, PRI515, R/G, HI e indices de estimacion de clorofila y carotenos demuestran ser buenos indicadores para la cuantificacion de dano moderado o severo causada por Verticilosis, lo que puede ser de utilidad para el diseno de estrategias de control de la Verticilosis a escala de parcela y de comarca. El olivo (Olea europaea L.) es el cultivo lenoso no tropical que ocupa mayor superficie en todo el mundo, con el 95% de la produccion mundial localizada en la cuenca mediterranea. Espana es el pais con mayor superficie de olivar del mundo con 2,6 MHa y aproximadamente el 39% de la produccion mundial. La Verticilosis, causada por el hongo de suelo Verticillium dahliae Kleb, constituye la principal amenaza para el olivar. Esta enfermedad afecta al olivo en todos los paises de tradicion olivarera y causa importantes perdidas de rendimiento y muerte de los arboles (Jimenez-Diaz y col., 2012). Este patogeno coloniza el sistema vascular de la planta, bloqueando el flujo del agua y finalmente induciendo estres hidrico (Van Alfen, 1989).Espana es el mayor productor de citricos de la Union Europea y el quinto a nivel mundial, con una produccion anual superior a cinco millones de toneladas (MAGRAMA, 2016). Este sector genera una gran cantidad de subproductos, con un elevado aporte energetico y alto contenido en pectinas y azucares, por lo que podrian usarse como materias primas en dietas para rumiantes. Sin embargo, los subproductos de citricos son muy diversos, por lo que su composicion quimica y valor nutritivo tambien puede ser variable (FEDNA, 2010; Feedipedia, 2016). El objetivo de este trabajo fue analizar la composicion quimica y la produccion de gas in vitro de subproductos de citricos producidos en nuestro pais.Resumen del trabajo presentado a las I Jornadas Cientificas del CIAL celebradas el 5 de junio de 2014 en Madrid.The Indian Summer Monsoon onset is one of the meteorological events most anticipated in the world. Due to its relevance for the population, the India Meteorological Department has dated the onset over the southern tip of the Indian Peninsula (Kerala) since 1901. The traditional method to date the onset was based in the judgment of skilled meteorologist and because of this, the method was considered subjective and not adequate for the study of long-term changes in the onset.Resumen del poster presentado a las I Jornadas Cientificas del CIAL celebradas el 5 de junio de 2014 en Madrid.Trabajo presentado en el European Workshop on Experimental and Behavioral Economics (EWEBE), organizado por la Universidad de Bologna los dias 26 y 27 de mayo de 2017 con el titulo: The impact of deliberative structures on voting behavio.--Trabajo presentado en los Bilkent Microeconomics Seminars organizados por la Bilkent University (Ankara, Turkey) el dia 29 de noviembre de 2017La compacidad del racimo es un rasgo de gran interes en la vid, dado que puede condicionar su calidad y rendimiento. Estos efectos se producen fundamentalmente a traves de su relacion, directa o indirecta, con la arquitectura del racimo, el comportamiento reproductivo y el grado o modo de exposicion de las bayas al ambiente. Respecto a los dos primeros, en un estudio previo, determinamos que tanto la longitud del raquis y sus primeras ramas como el numero de bayas por racimo son los caracteres con mayor incidencia en la compacidad entre los analizados. Ahora bien, el estudio de la base genetica responsable del numero de bayas requiere de su diseccion previa, dado que es una variable secundaria, resultado del numero de flores por inflorescencia y de la tasa de cuajado o conversion de flor en fruto. En este trabajo se presenta un estudio preliminar en el que se han caracterizado fenotipicamente 104 variedades de vid para estudiar su comportamiento reproductivo. Asi, sobre una base amplia de diversidad varietal, se han estimado variables como el numero de flores, la tasa de cuajado, corrimiento, millerandage, etc., evaluando su posible relacion con la compacidad del racimo. Este trabajo, una vez completado durante varios anos, permitira conocer la diversidad existente para el comportamiento reproductivo en la vid, asi como, de forma mas concreta, abordar la caracterizacion de los procesos geneticos responsables de las diferencias fenotipicas detectadas para el comportamiento reproductivo y sus elementos individuales y para la compacidad del racimo, identificando los posibles genes involucrados en dichos procesos.Trabajo presentado en la 11th International Conference on Grapevine Breeding and Genetics, celebrada en Pekin del 29 de julio al 2 de agosto de 2014.The Polarimetric and Helioseismic Imager (PHI) is the first deep-space solar spectropolarimeter, on-board the Solar Orbiter (SO) space mission. It faces: stringent requirements on science data accuracy, a dynamic environment, and severe limitations on telemetry volume. SO/PHI overcomes these restrictions through on-board instrument calibration and science data reduction, using dedicated firmware in FPGAs. This contribution analyses the accuracy of a data processing pipeline by comparing the results obtained with SO/PHI hardware to a reference from a ground computer. The results show that for the analysed pipeline the error introduced by the firmware implementation is well below the requirements of SO/PHI.This work has been financed within the framework of the projects P/309307 Arqueoastronomia of the IAC, and Orientatio ad sidera III (AYA2011-26759) of the Spanish MINECO. ACGG is Ramon y Cajal researcher of the MINECO.Este trabajo ha sido financiado por el proyecto RECUPERA 2020 del MINECO y EU-FEDER; y por el proyecto TRANS·FORMA (PP.TRA.TAA201600.9). La investigacion realizada por E. Rodriguez ha sido realizada gracias al programa DOC-INIA (INIA-FEDER).Trabajo presentado en el XXIV Congreso Iberoamericano de Catalisis (CICat 2014), “Catalisis para Biorrefineria”, celebrado en Medellin del 11 al 13 de septiembre de 2014.2 paginas y 1 tabla.- Trabajo presentado en el IX Congreso de Mejora Genetica de Plantas celebrado en Murcia entre el 18 y el 20 de septiembre de 2018.3 paginas, 1 tabla, 1 figura.--Trabajo presentado a las XVI Jornadas sobre Produccion Animal AIDA (Zaragoza, 19 al 20 de mayo, 2015).Poster presentado en el XVI International Clay Conference from the Oceans to Space Granada, Spain, July 17-21, (2017)XIII Reunion Iberica de Algas Toxicas y Biotoxinas Marinas (REDIBAL 2018) - XIII Reunion Iberica de Fitoplancton Toxico - XIII Iberian Toxic Algae and Marine Biotoxins Meeting, VI Simposio Internacional de Ciencias del Mar - VI International Symposium of Marine Sciences (ISMS 2018), 20- 22 June 2018, Vigo.-- 2 pagesThis research was financed with project AGL2011-22783 granted by the Spanish Ministry of Economy and Competiveness. M.C. Soto was supported by a CONACYT (Mexican Council of Sciences and Technology) doctoral fellowship.This work was supported by the Spanish Ministerio de Economia y Competitividad (grants CGL2012-36251 and CGL2015-64727-P to X.B., and BFU2010-15906 to JLM), and Catalan Government (2014 SGR 619). The research has also benefited from FEDER funds.Trabajo presentado en el IX International Symposium on Grapevine Physiology and Biotechnology, celebrado en La Serena (Chile) del 21 al 26 de abril de 2013.5 paginas, 4 figuras, 1 tabla.--Trabajo presentado a las: XLI Jornadas Cientificas y XVII Jornadas Internacionales de Ovinotecnia y Caprinotecnia. (Talavera de la Reina, Toledo, Espana, 14-16 septiembre, 2016).Se presenta una revisión histórica de los equinodermos Ordovícicos de Portugal. Todos los registros conocidos hasta el momento pertenecen al Ordovícico Medio-Superior (Darriwiliense-Katiense) y las asociaciones presentan gran diversidad aunque en general los taxones están poco o mal conocidos. La mayor parte de los yacimientos se concentran en las regiones de Amêndoa-Mação y Buçaco, aunque también existen citas de equinodermos en Arouca, Dornes, Moncorvo y Valongo. Por último se comentan futuras líneas de trabajo que ayudarán a comprender mejor estas faunas, perfeccionar y completar el conocimiento del registro fósil del grupo en el Ordovícico centroibérico. Ordovician echinoderms from Portugal: state of the artResumen del trabajo presentado al 37o Convegno Internazionale dei Docenti della Rappresentazione de la Unione Italiana Disegno (UDI), celebrado en Torino (Italia) del 17 al 19 de septiembre de 2015.Trabajo presentado al Danish Days on Caloric Materials and Devices, celebrado en Roskilde (Dinamarca) del 2 al 3 de octubre de 2017.Trabajo presentado en el ImageNano 2015 (Bringing together Nanoscience and Nanotechnology), celebrado en Bilbao del 10 al 13 de marzo de 2015.Este estudio ha sido financiado por el proyecto AGL-2012-40172-C02-01 del Ministerio de Economia y Competitividad (MINECO). AEF agradece al MINECO la financiacion de su contrato de investigacion (FPI).We present a detailed study of a X -ray selected sample of 5 submillimeter bright QSOs at