Michela Re

Chemical restraint and anaesthetic effects of a tiletamine-zolazepam/ketamine/detomidine combination in cattle.

The immobilisation and anaesthesia of free-ranging cattle requires the administration of appropriate drugs in small volume via rifle or dart. The objective of this randomised controlled study was to test the capacity of a concentrated combination of tiletamine-zolazepam (TZ), ketamine (K) and detomidine (D) (TZKD) to immobilise/anaesthetise calves. Following administration of low, medium and high doses of TZKD to six healthy animals IM, the time-of-onset and duration of anaesthesia were recorded, in addition to standard cardio-respiratory parameters. Two noxious stimuli were applied to assess the analgesic effect of the combination. TZKD produced a dose-dependent anaesthetic action associated with respiratory depression and moderate hypoxaemia. Total recumbency lasted from 1h (with low dose) to 2h (with medium and high doses). The findings indicate that TZKD induces anaesthesia in calves, suitable not only for animal immobilisation, but also to carry out minor surgical procedures with or without additional local analgesia. Respiratory depression was the most severe side-effect and careful patient monitoring is recommended when using this drug combination.

Comparison of sedation scores and propofol induction doses in dogs after intramuscular administration of dexmedetomidine alone or in combination with methadone, midazolam, or methadone plus midazolam.

The objectives of this study were to determine: (1) the sedative effects of dexmedetomidine in combination with methadone, midazolam, or both, and (2) the propofol dose required to achieve endotracheal intubation in healthy dogs. Seven healthy Beagle dogs were included in a prospective experimental, crossover, randomised and masked design. All dogs received four treatments IM, with at least 1 week between sessions, as follows: dexmedetomidine 5 µg/kg (D) alone, or combined with methadone 0.3 mg/kg (DMe), midazolam 0.3 mg/kg (DMi), or both (DMeMi). The degree of sedation was evaluated using a numerical scale (maximum 15 points). The dose of propofol required for intubation was also calculated for each group. Recovery time and quality were determined. Statistical analysis was performed using parametric (ANOVA) and nonparametric tests (Friedman, Cochran Q), as appropriate. The degree of sedation obtained with DMe and DMeMi (13, [7-14]; 13, [6-14], respectively) was significantly higher than in the control group (2, [1-4]; P = 0.023, P = 0.006, respectively). The required dose of propofol was lower in all groups (DMi, 1.5 ± 0.5 mg/kg, P = 0.002; DMe, 1.2 ± 0.5 mg/kg, P <0.001; DMeMi, 0.9 ± 0.3 mg/kg) than in the control group (2.9 ± 0.9 mg/kg; P <0.001). Recovery quality was not different between groups (P = 0.137). In healthy dogs, the addition of midazolam did not enhance the sedative effects of dexmedetomidine or a dexmedetomidine-methadone combination at the doses studied, and propofol requirements were reduced. The sedative effect of dexmedetomidine was enhanced with methadone, and the required dose of propofol was reduced.

Ultrasound-Guided Nerve Block Anesthesia

Superficial nerves can be visualized through ultrasonography in the cattle and facilitate local anesthetic disposition around nerve structures. Expected advantages include a higher successful rate of nerve block improving the degree and duration of the block. Among others, conduction nerves of clinical interest in cattle include the paravertebral nerves, nerves of the epidural space, the brachial plexus, and the sciatic and femoral nerves, and nerves of the head.

Ultrasound-guided anaesthetic blockade of the pelvic limb in calves

This study aimed to describe a suitable acoustic window to facilitate access to the sciatic and femoral nerves in calves and to study the effects of their blockade with local anaesthetics. The neuroanatomical and ultrasound (US) study was performed on the cadavers of 10 calves, and the effects of 2% lidocaine with epinephrine (0.2 mL/kg) were determined in five healthy calves. The sciatic nerve in the cadavers was easily visualised as a hyperechoic band distal to the femoral greater trochanter and caudal to the femoral shaft. The femoral nerve in the cadavers was not easily identified, and was visualised as a hyperechoic oval structure situated immediately medial to the psoas major muscle and lateral to the femoral artery. The sciatic nerve was stained by methylene blue, injected under US guidance, in 9/10 cases, and the femoral nerve was stained in 6/10 cases. Sciatic nerve blockade under US guidance produced adduction of the limb with metatarsophalangeal joint flexion, while the femoral nerve blockade produced reduced weight bearing. The sciatic nerve blockade produced a reduced response to the noxious stimulus, mainly in the phalanges, proximal and distal metatarsus, tarsus and tibia and, following the femoral nerve blockade, in the medial subarea of the femur. However, femoral nerve blockade produced a more variable degree of blockade. In conclusion, US -guided anaesthetic blockade of the sciatic nerve in calves may be considered for surgery in the distal pelvic limb, although further studies are necessary to determine its clinical application.

Safety and efficacy of the bumped kinase inhibitor BKI-1553 in pregnant sheep experimentally infected with Neospora caninum tachyzoites

Neospora caninum is one of the main causes of abortion in cattle, and recent studies have highlighted its relevance as an abortifacient in small ruminants. Vaccines or drugs for the control of neosporosis are lacking. Bumped kinase inhibitors (BKIs), which are ATP-competitive inhibitors of calcium dependent protein kinase 1 (CDPK1), were shown to be highly efficacious against several apicomplexan parasites in vitro and in laboratory animal models. We here present the pharmacokinetics, safety and efficacy of BKI-1553 in pregnant ewes and foetuses using a pregnant sheep model of N. caninum infection. BKI-1553 showed exposure in pregnant ewes with trough concentrations of approximately 4 μM, and of 1  μM in foetuses. Subcutaneous BKI-1553 administration increased rectal temperatures shortly after treatment, and resulted in dermal nodules triggering a slight monocytosis after repeated doses at short intervals. BKI-1553 treatment decreased fever in infected pregnant ewes already after two applications, resulted in a 37–50% reduction in foetal mortality, and modulated immune responses; IFNγ levels were increased early after infection and IgG levels were reduced subsequently. N. caninum was abundantly found in placental tissues; however, parasite detection in foetal brain tissue decreased from 94% in the infected/untreated group to 69–71% in the treated groups. In summary, BKI-1553 confers partial protection against abortion in a ruminant experimental model of N. caninum infection during pregnancy. In addition, reduced parasite detection, parasite load and lesions in foetal brains were observed.

Influence of dose and route of administration on the outcome of infection with the virulent Neospora caninum isolate Nc-Spain7 in pregnant sheep at mid-gestation

Experimental infections in pregnant sheep have been focused on studying the effect of the time of challenge on the outcome of N. caninum infection, whereas the impact of the dose and route of challenge has not been studied in depth. Therefore, clinical outcome, immune responses, parasite detection and burden, and lesion severity in placental tissues and foetal brains were investigated in 90-day-pregnant sheep inoculated intravenously with 105 (G1), 104 (G2), 103 (G3), or 102 (G4) tachyzoites or subcutaneously with 104 (G5) tachyzoites of the virulent Nc-Spain7 isolate and an uninfected group (G6). Comparing challenge doses, G1 was the only group that had 100% abortion. Likewise, IFNγ levels in G1 increased earlier than those in other intravenously infected groups, and IgG levels on day 21 post-infection (pi) were higher in G1 than those in other intravenously infected groups. Concerning vertical transmission, G1 shows a higher parasite burden in the foetal brain than did G2 and G3. Comparing routes of administration, no differences in foetal survival rate or parasite load in the foetal brain were found. Although G2 had higher IFNγ levels than G5 on day 10 pi, no differences were found in humoral immune responses. Because the outcome after intravenous infection with 105 tachyzoites was similar to that observed after intravenous infection with 106 tachyzoites used in a previous work (100% abortion and vertical transmission), we conclude that it may be reasonable to use 105 tachyzoites administered by the intravenous route in further experiments when assessing drugs or vaccine candidates.

Pharmacokinetics, safety and efficacy of Bumped Kinase Inhibitor (BKI) 1553 in a pregnant sheep model of neosporosis

This work was supported by Community of Madrid (Grupo Estrategico 2000-2003), NIH, grant R01CA77575, and SAF 2001-2245.The transition step from the p3-dAMP initiation complex to the first elongated products, p3-(dAMP)2 and p3-(dAMP)3, requires a dATP concentration higher than that needed for the initiation reaction or for the further elongation of the p3-(dAMP)3 complex. The elongation in phi 29 DNA-protein p3 replication in vitro was strongly inhibited by salt. Under inhibitory salt concentration, the viral protein p6 greatly stimulated phi 29 DNA-protein p3 replication. The effect of protein p6 was not on the rate of elongation but on the amount of elongated product, stimulating the transition from initiation to formation of the first elongation products.Trabajo presentado en 44th Annual Meeting Society for Neuroscience, celebrado en Washington, DC (USA) del 15 al 19 de noviembre de 2014Recent studies have demonstrated that cytochrome c plays an important role in cell death. In the present study, we report that teniposide and various other chemotherapeutic agents induced a dose-dependent increase in the expression of the mitochondrial respiratory chain proteins cytochrome c, subunits I and IV of cytochrome c oxidase, and the free radical scavenging enzyme manganous superoxide dismutase. The teniposide-induced increase of cytochrome c was inhibited by cycloheximide, indicating new protein synthesis. Elevated cytochrome c levels were associated with enhanced cytochrome c oxidase-dependent oxygen uptake using TMPD/ascorbate as the electron donor, suggesting that the newly synthesized proteins were functional. Cytochrome c was released into the cytoplasm only after maximal levels had been reached in the mitochondria, but there was no concomitant decrease in mitochondrial membrane potential or caspase activation. Our results suggest that the increase in mitochondrial protein expression may play a role in the early cellular defense against anticancer drugs.Supported by Grant GM-08041 from the National Institutes of Health, United States Public Health Service.The results presented in this paper indicate that the phi 29 DNA polymerase is the only enzyme required for efficient synthesis of full length phi 29 DNA with the phi 29 terminal protein, the initiation primer, as the only additional protein requirement. Analysis of phi 29 DNA polymerase activity in various in vitro DNA replication systems indicates that two main reasons are responsible for the efficiency of this minimal system: 1) the phi 29 DNA polymerase is highly processive in the absence of any accessory protein; 2) the polymerase itself is able to produce strand displacement coupled to the polymerization process. Using primed M13 DNA as template, the phi 29 DNA polymerase is able to synthesize DNA chains greater than 70 kilobase pairs. Furthermore, conditions that increase the stability of secondary structure in the template do not affect the processivity and strand displacement ability of the enzyme. Thus, the catalytic properties of the phi 29 DNA polymerase are appropriate for a phi 29 DNA replication mechanism involving two replication origins, strand displacement and continuous synthesis of both strands. The enzymology of phi 29 DNA replication would support a symmetrical model of DNA replication.Aided by grants from the National Institutes of Health U.S. Public Health Service, and E. I. Du Pont de Neumours and Company, Inc.This work was supported in part by NRSA, National Institutes of Health Grants NS09463 and NS32501 and from National Science Foundation Grant 9310965.We have recently developed a new method to detect and characterize single base substitutions in transcribed genes which is based on the ability of RNAse A to recognize and cleave single base mismatches in RNA:RNA heteroduplexes. The RNAse A misrnatch cleavage assay was applied to screen human colon carcinoma cell lines and primary tumors for the presence of mutant e-X-ras oncogenes. We have determined that the mutant e-X-ras allele is overexpressed and amplified relative to the normal in the SX-CO-l human colon carcinoma cell lineo The oncogene mutation has been characterized by this method as a glycine to valine substitution at codon 12 of the e-X-ras gene. This result was confirmed by cloning and sequencing. We have previously reported that about 40% of primary human colon tumors contain e-X-ras genes mutant at codon 12 (Forrester et al, Nature 327: 298, 1987). We report here the characterization by molecular cloning and sequencing of the mutation in the e-X-ras oneogene from two of these tumors (tumors 3 and 28). We also describe the histopathologieal eharaeterization of these two tumors and demonstrate, by Southern blot hybridization of NIH3T3 transformants, the simultaneous presenee of mutant e-X-ras and N-ras oncogenes in villous adenoma 28. Our results provide evidence for the frequent assoeiation of ras somatie mutational aetivation in the early stages of tumor development in this common type of human eaneer.Aided by Grants AM-01845, AM-08953, and l-Sol-FR-05099 from the National Institutes o f Health, United States Public Health Service, and E. I. Du Pont de Nemours and Company, Inc. A preliminary report o f this work was presented at the Second Meeting o f the Federation o f European Biochemical Societies (symposium on “Ribonucleic Acid-Structure and Function”), Vienna, April 21 to 24, 1965.1 pagina.-- Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (Madrid, Spain. 11-14 October ,2017).Supported by Grant GM-08041 from the National Institutes of Health, United States Public Health Service.Resumen del trabajo presentado al XXXIII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular celebrado en Cordoba del 14 al 17 de septiembre de 2010.This article describes the expression pattern and functional analysis of Lazarillo, a novel cell surface glycoprotein expressed in the embryonic grasshopper nervous system, and a member of the lipocalin family. Lazarillo is expressed by a subset of neuroblasts, ganglion mother cells and neurons of the central nervous system, by all sensory neurons of the peripheral nervous system, and by a subset of neurons of the enteric nervous system. It is also present in a few non neuronal cells associated mainly with the excretory system. A monoclonal antibody raised against Lazarillo perturbs the extent and direction of growth of identified commissural pioneer neurons. We propose that Lazarillo is the receptor for a midline morphogen involved in the outgrowth and guidance of these neurons.Poster presentado al Annual Biomedical Research Conference for Minority Students celebrado en California (US) del 7 al 10 de noviembre de 2012.The phage phi 29 regulatory protein p4 activates the late promoter A3 by stabilizing the binding of Bacillus subtilis RNA polymerase (RNAP) as a closed complex. Interaction between the two proteins occurs through amino acid Arg120 in protein p4 and the C-terminal domain of the RNAP alpha subunit (alpha-CTD). In addition to its role as activator of the late transcription, protein p4 represses early transcription from the A2b and A2c promoters, that are divergently transcribed. Binding of p4 to its recognition site at the A3 promoter displaces the RNAP from promoter A2b, both by steric hindrance and by the curvature induced upon p4 binding. At the A2c promoter, the RNAP cooperates with p4 binding in such a way that promoter clearance is prevented. Interestingly, amino acid Arg120 in p4 and the alpha-CTD in B. subtilis RNAP are involved in the interactions that lead to transcription repression at promoter A2c. To investigate how this interaction leads to activation at PA3 and to repression at PA2c, mutant promoters were constructed. In the absence of a -35 consensus box for sigma A-RNAP activation was observed, while in its presence repression occurred. The results support the idea that overstabilization of RNAP at the promoter over a threshold level leads to repression.Resumen del poster presentado al XXXIII Congreso de la Sociedad Espanola de Bioquimica y Biologia Molecular celebrado en Cordoba del 14 al 17 de septiembre de 2010.Formalin-fixed paraffin-embedded tissue specimens obtained by fine needle aspiration of pancreatic masses from 47 patients were examined retrospectively for cytology and the presence of mutant c-K-ras oncogenes. Point mutations of c-K-ras in codon 12 were detected by RNA-DNA RNAse A mismatch cleavage after in vitro DNA amplification of the cellular c-K-ras sequences by the polymerase chain reaction. Of the 36 patients with pancreatic adenocarcinoma, mutant c-K-ras oncogenes were detected in 18 of 25 (72%) with malignant cytologies, 2 of 8 (25%) with atypical cytologies, and 0 of 3 with benign aspiration cytologies. The remaining 11 patients without pancreatic adenocarcinomas did not have mutant c-K-ras genes detectable by the assay. The diagnosis of pancreatic adenocarcinoma was based upon clinical follow-up. The presence of mutant c-K-ras oncogenes did not significantly affect survival in the patients studied. Mutant c-K-ras genes were found at the time of initial clinical presentation in the majority of pancreatic adenocarcinomas, suggesting an important role of the mutation in oncogenesis. In conjunction with cytology, our approach represents an application for cancer diagnosis at the molecular genetic level.Calorie restriction (CR) has been shown to decrease reactive oxygen species (ROS) production and retard aging in a variety of species. It has been proposed that alterations in membrane saturation are central to these actions of CR. As a step towards testing this theory, mice were assigned to 4 dietary groups (control and 3 CR groups) and fed AIN-93G diets at 95 % (control) or 60 % (CR) of ad libitum for 8 months. To manipulate membrane composition, the primary dietary fats for the CR groups were soybean oil (also used in the control diet), fish oil or lard. Skeletal muscle mitochondrial lipid composition, proton leak, and H(2)O(2) production were measured. Phospholipid fatty acid composition in CR mice was altered in a manner that reflected the n-3 and n-6 fatty acid profiles of their respective dietary lipid sources. Dietary lipid composition did not alter proton leak kinetics between the CR groups. However, the capacity of mitochondrial complex III to produce ROS was decreased in the CR lard compared to the other CR groups. The results of this study indicate that dietary lipid composition can influence ROS production in muscle mitochondria of CR mice. It remains to be determined if lard or other dietary oils can maximize the CR-induced decreases in ROS production.To investigate the relationship between RNA folding and ribozyme catalysis, we have carried out a detailed kinetic analysis of four structural derivatives of the hairpin ribozyme. Optimal and suboptimal (wild-type) substrate sequences were studied in conjunction with stabilization of helix 4, which supports formation of the catalytic core. Pre-steady-state and steady-state kinetic studies strongly support a model in which each of the ribozyme variants partitions between two major conformations leading to active and inactive ribozymez substrate complexes. Reaction rates for cleavage, ligation, and substrate binding to both ribozyme conformations were determined. Ligation rates (3 min 21 ) were typically 15-fold greater than cleavage rates (0.2 min 21 ), demonstrating that the hairpin ribozyme is an efficient RNA ligase. On the other hand, substrate binding is very rapid (k on 5 4 3 10 8 M 21 min 21 ), and the ribozymez substrate complex is very stable (K D < 25 pM ;k off < 0.01 min 21 ). Stabilization of helix 4 increases the proportion of RNA molecules folded into the active conformation, and enhances substrate association and ligation rates. These effects can be explained by stabilization of the catalytic core of the ribozyme. Rigorous consideration of conformational isomers and their intrinsic kinetic properties was necessary for development of a kinetic scheme for the ribozyme-catalyzed reaction.The human integrin VLA (very late activation antigens)-4 (CD49d/CD29), the leukocyte receptor for both the CS-1 region of plasma fibronectin (Fn) and the vascular cell surface adhesion molecule-1 (VCAM-1), also mediates homotypic aggregation upon triggering with specific anti-VLA-4 monoclonal antibody (mAb). Epitope mapping of this integrin on the human B-cell line Ramos, performed with a wide panel of anti-VLA-4 mAb by both cross-competitive cell binding and protease sensitivity assays, revealed the existence of three topographically distinct epitopes on the alpha 4 chain, referred to as epitopes A-C. By testing this panel of anti-VLA-4 mAb for inhibition of cell binding to both a 38-kDa Fn fragment containing CS-1 and to VCAM-1, as well as for induction and inhibition of VLA-4 mediated homotypic cell adhesion, we have found overlapping but different functional properties associated with each epitope. Anti-alpha 4 mAb recognizing epitope B inhibited cell attachment to both Fn and VCAM-1, whereas mAb against epitope A did not block VCAM-1 binding and only partially inhibited binding to Fn. In contrast, mAb directed to epitope C did not affect cell adhesion to either of the two VLA-4 ligands. All mAb directed to site A, as well as a subgroup of mAb recognizing epitope B (called B2), were able to induce cell aggregation, but this effect was not exerted by mAb specific to site C and by a subgroup against epitope B (called B1). Moreover, although anti-epitope C and anti-epitope B1 mAb did not trigger aggregation, those mAb blocked aggregation induced by anti-epitope A or B2 mAb. In addition, anti-epitope A mAb blocked B2-induced aggregation, and conversely, anti-epitope B2 mAb blocked A-induced aggregation. Further evidence for multiple VLA-4 functions is that anti-Fn and anti-VCAM-1 antibodies inhibited binding to Fn or to VCAM-1, respectively, but did not affect VLA-4-mediated aggregation. In summary, we have demonstrated that there are at least three different VLA-4-mediated adhesion functions, we have defined three distinct VLA-4 epitopes, and we have correlated these epitopes with the different functions of VLA-4.Lazarillo, a protein recognized by the monoclonal antibody 10E6, is expressed by a subset of neurons in the developing nervous system of the grasshopper. It is a glycoprotein of 45x10(3) M(r) with internal disulfide bonds and linked to the extracellular side of the plasma membrane by a glycosylphosphatidylinositol moiety. Peptide sequences obtained from affinity purified adult protein were used to identify an embryonic cDNA clone, and in situ hybridizations confirmed that the distribution of the Lazarillo mRNA paralleled that of the monoclonal antibody labeling on embryos. Sequence analysis defines Lazarillo as a member of the lipocalin family, extracellular carriers of small hydrophobic ligands, and most related to the porphyrin- and retinol-binding lipocalins. Lazarillo is the first example of a lipocalin anchored to the plasma membrane, highly glycosylated, and restricted to a subset of developing neurons.Trabajo presentado al Annual Biomedical Research Conference for Minority Students celebrada en Nashville (US) del 13 al 16 de noviembre de 2013.A cDNA has been isolated from human hippocampus that appears to encode a novel Na(+)-dependent, Cl(-)-independent, neutral amino acid transporter. The putative protein, designated SATT, is 529 amino acids long and exhibits significant amino acid sequence identity (39-44%) with mammalian L-glutamate transporters. Expression of SATT cDNA in HeLa cells induced stereospecific uptake of L-serine, L-alanine, and L-threonine that was not inhibited by excess (3 mM) 2-(methylamino)-isobutyric acid, a specific substrate for the System A amino acid transporter. SATT expression in HeLa cells did not induce the transport of radiolabeled L-cysteine, L-glutamate, or related dicarboxylates. Northern blot hybridization revealed high levels of SATT mRNA in human skeletal muscle, pancreas, and brain, intermediate levels in heart, and low levels in liver, placenta, lung, and kidney. SATT transport characteristics are similar to the Na(+)-dependent neutral amino acid transport activity designated System ASC, but important differences are noted. These include: 1) SATTs apparent low expression in ASC-containing tissues such as liver or placenta; 2) the lack of mutual inhibition between serine and cysteine; and 3) the lack of trans-stimulation. SATT may represent one of multiple activities that exhibit System ASC-like transport characteristics in diverse tissues and cell lines.

Experimental infection with Besnoitia besnoiti tachyzoites in calves and young bulls

La enfermedad de Lafora es un tipo de epilepsia mioclonica progresiva de herencia autosomica recesiva causada por la mutacion de uno de los siguientes genes: EPM2A, que codifica para la fosfatasa de especificidad dual laforina, y EPM2B, que codifica para la E3 ubicuitina ligasa malina. Ambas proteinas participan en las mismas rutas fisiologicas a traves de la formacion de un complejo funcional en el que laforina recluta de manera especifica sustratos para que sean ubicuitinados por malina. Sin embargo, muchos aspectos relativos al funcionamiento de este complejo estan aun por definir. En el presente trabajo, hemos analizado el mecanismo de dimerizacion de laforina, descubriendo que la cisteina 329 juega un papel fundamental en el proceso de dimerizacion, ya que su mutacion da lugar a una forma exclusivamente monomerica que mantiene la capacidad catalitica y la capacidad de formar un complejo funcional con malina. Por otro lado, hemos definido que el complejo laforina-malina emplea a la enzima conjugadora de ubicuitina UBE2N para promover la ubicuitinacion de sus sustratos con cadenas unidas en K63. El adaptador de la autofagia selectiva p62 tambien interacciona con el complejo laforina-malina, estimulando su capacidad de ubicuitinacion y siendo ubicuitinado el mismo en el proceso. Todos ellos colocalizan en regiones donde esta teniendo lugar la autofagia. Ademas, el complejo laforina-malina participa en la regulacion de este proceso uniendo al nucleo central beclina1-Vps34-Vps15, promoviendo su ubicuitinacion, asi como la de reguladores posteriores como Atg14L y UVRAG. El complejo laforina-malina esta regulado por componentes de la maquinaria de autofagia, como Bcl-2, que inhibe su capacidad de ubicuitinacion, o TRAF6, que promueve la ubicuitinacion de laforina. Sin embargo, el complejo laforina-malina no se recluta a la mitocondria para su degradacion por autofagia ni en condiciones normales ni tras producirse dano mitocondrial. Del mismo modo, la migracion de parkina a la mitocondria para inducir su degradacion no esta afectada ni en ausencia ni en sobreexpresion de laforina y malina. En conjunto, hemos definido algunos aspectos relevantes respecto al mecanismo por el que actuan laforina y malina, y por tanto mejorado nuestra comprension sobre como pueda estar causada la patologia.290 Pags.- Figs.- Tabls. Tesis Univ. Zaragoza, Departamento de Geografia y Ordenacion del Territorio. Dirigida por los Drs. Santiago Begueria (EEAD-CSIC) y Sergio M. Vicente-Serrano (IPE-CSIC). Bajo Creative Commons License Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0).We define a clause tableau calculus for MaxSAT, prove its soundness and completeness, and describe a tableau-based algorithm for MaxSAT. Given a multiset of clauses ϕ, the algorithm computes both the minimum number of clauses that can be falsified in ϕ, and an optimal assignment. We also describe how the algorithm can be extended to solve weighted MaxSAT and weighted partial MaxSAT.Grupo 5: Valor social.-- Programa de investigacion para la conservacion preventiva y regimen de acceso de la cueva de Altamira (2012-2014).Automated planning has proven to be useful to solve problems where an agent has to maximize a reward function by executing actions. As planners have been improved to salve more expressive and difficult problems, there is an increasing interest in using planning to improve efficiency in robotic tasks. However, planners rely on a domain model, which has to be either handcrafted or learned. Although learning domain models can be very costly, recent approaches provide generalization capabilities and integrate human feedback to reduce the amount of experiences required to learn. In this thesis we propase new methods that allow an agent with no previous knowledge to solve certain problems more efficiently by using task planning. First, we show how to apply probabilistic planning to improve robot performance in manipulation tasks (such as cleaning the dirt or clearing the tableware on a table). Planners obtain sequences of actions that get the best result in the long term, beating reactive strategies. Second, we introduce new reinforcement learning algorithms where the agent can actively request demonstrations from a teacher to learn new actions and speed up the learning process. In particular, we propase an algorithm that allows the user to set the minimum quality to be achieved, where a better quality also implies that a larger number of demonstrations will be requested . Moreover, the learned model is analyzed to extract the unlearned or problematic parts of the model. This information allow the agent to provide guidance to the teacher when a demonstration is requested, and to avoid irrecoverable errors. Finally, a new domain model learner is introduced that, in addition to relational probabilistic action models, can also learn exogenous effects. This learner can be integrated with existing planners and reinforcement learning algorithms to salve a wide range of problems. In summary, we improve the use of learning and task planning to salve unknown tasks. The improvements allow an agent to obtain a larger benefit from planners, learn faster, balance the number of action executions and teacher demonstrations, avoid irrecoverable errors, interact with a teacher to solve difficult problems, and adapt to the behavior of other agents by learning their dynamics. All the proposed methods were compared with state-of-the-art approaches, and were also demonstrated in different scenarios, including challenging robotic tasks.L’estudi del sistema d’alimentacio d’hidrogen es un dels camps de recerca necessaris per a allargar la vida util de les piles de combustible PEM. Estudis anteriors han demostrat que la vida util i l’eficiencia d’aquestes piles milloren amb l’us d’un sistema de recirculacio d’hidrogen. En aquest treball s’ha dissenyat i fabricat un ejector per a ser implementat a l’estacio de proves 4 del laboratori de control de piles de combustible de l’IRI amb la finalitat d’analitzar l’efecte que els sistemes de recirculacio d’hidrogen basats en ejectors tenen sobre aquestes piles. En el disseny de la geometria d’un ejector s’han de considerar diferents parametres que nomes es poden estudiar mitjancant CFD (Computational Fluid Dynamics). Per tant, s’ha desenvolupat un model 2D axisimetric utilitzant el model de turbulencia k-e estandard per a resoldre les equacions de Navier-Stokes amb una mitjana de Favre. El model s’ha utilitzat per a dissenyar i fabricar un ejector experimental amb la finalitat de validar-lo. Els resultats obtinguts mostren que el model es capac d’obtenir els fluxos massics que apareixen en un ejector per a diferents condicions operatives. Un cop validat el model, s’ha fet servir per a fer un estudi parametric per a trobar els parametres geometrics optims que ha de tenir l’ejector dissenyat per a l’estacio de proves 4.Trabajo presentado en el 10th International Symposium on the Cretaceous, celebrado en Viena (Austria) del 21 al 26 de agosto de 2017Este trabajo ha sido parcialmente financiado por el proyecto CGL2009-11316 y por una beca de Formacion de Profesorado Universitario del Ministerio de educacion AP2007-03633.Trabajo presentado en el XXIV Congreso Iberoamericano de Catalisis, celebrado en Medellin (Colombia) del 14 al 19 de septiembre de 2014.Trabajo presentado en el 33rd International Meeting of Sedimentology y 16eme Congres Francais de Sedimentologie (2017), celebrado en Toulouse (Francia), del 10 al 12 de octubre de 2017Trabajo presentado en la XXV Reunion Bienal de Quimica Organica celebrada en Alicante del 4 al 6 de junio de 2014.Resumen del trabajo presentado a las II Jornadas Cientificas CIAL-Forum, celebradas durante los dias 16 y 17 de noviembre de 2016 en el Instituto de Investigacion en Ciencias de la Alimentacion (CIAL).Esta reflexion analiza el paralelismo entre el origen del discurso patrimonial autorizado y la precarizacion del trabajo femenino. Ambos surgen con el capitalismo y son dos de sus pilares. Plantea que el cambio simbolico que implica la igualdad legal entre hombres y mujeres supone un cambio profundo en las estructuras sociales y que este cambio ofrece serias resistencias, que se generan no solo por la necesidad de las instancias de poder de seguir ocupandolo sino porque implican cambios en la identidad de los grupos y de los individuos, con la angustia que ello genera. La idea de patrimonio cultural esta directamente implicada en estos cambios puesto que la incorporacion de las mujeres a el implica una variacion sustancial en el propio concepto. Sugiere que para avanzar en este cambio es necesario comprender que el “que” y el “como” se hacen las cosas estan necesariamente imbricados.Presentacion para el 50th European Marine Biology Symposium, 21-25 September 2015, Helgoland, Germany.-- 15 pagesResumen del trabajo presentado al XVIII Scientific Meeting of the Spanish Society of Chromatography and Related Techniques (SECyTA), celebrado en Granada del 2 al 4 de octubre de 2018.Resumen del trabajo presentado al 42st Annual Association for Research in Otolaryngology (ARO) MidWinter Meeting, celebrado en Baltimore, Maryland (USA) del 9 al 13 de febrero de 2019.Partial financial support has been received from the Agencia Estatal de Investigacion (AEI, MCI, Spain) and Fondo Europeo de Desarrollo Regional (FEDER, UE), under Project PACSS (RTI2018-093732-B-C21/C22) and the Maria de Maeztu Program for units of Excellence in R&D (MDM-2017-0711). A.F.P. acknowledges support by the Formacion de Profesorado Universitario (FPU14/00554) program of Ministerio de Educacion, Cultura y Deportes (MECD) (Spain).Memoria presentada para optar al titulo de Doctor otorgado por la Universidad de Cantabria por Biuse Casaponsa Gali y que ha sido realizada en el Instituto de Fisica de Cantabria.Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (Madrid, Spain. 11-14 October 2017).3 paginas, 1 tabla , 1 figura.--Trabajo presentado a las XVII Jornadas sobre Produccion Animal AIDA (Zaragoza, 30 y 31 de mayo, 2017).301 paginas.-- 392 referencias.-- Memoria para optar al titulo de Doctora en Farmacia por la Universidad de SevillaTrabajo presentado en el ANQUE.ICCE.BIOTEC - Congress on Chemistry, Chemical Engineering and Biotechnology, celebrado en Madrid del 01 al 04 de julio de 2014.Towards an integrated evaluation of the Spanish journals on Social and Human Sciences. The journals of Psychology. The present work is part of an ampler project about Spanish journals evaluation of Social y Human Sciences. First, the results of the evaluation of the Spanish journals of Psychology are presented. These results are obtained from a survey sent to university professors in order to evaluate the journals to establish hierarchized relations in each area of knowledge. Second, a study of the references cited from a selection of journals is realized. Finally, an integrated table of valuation of the journals is presented. In order to facilitate the task a data base is elaborated. Among the obtained conclusions it is particularly interested the fact that while the professors declare to publish mainly in Spanish journals, they preferredly cite foreign journals. On the other hand, it is indicated that the different methods to evaluate a journal are complementary and it is important to know the used method in each case.Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (11-14 October 2017 - Madrid, Spain).Trabajo presentado en la ed. anual 2017 del congreso nacional BIOTEC organizado por la Sociedad Espanola de Biotecnologia en el Campus La Merced de la Universidad de Murcia durante los dias 18 a 21 de junio de 2017.Memoria presentada para optar al grado de Doctor en Ciencias, Seccion Quimicas, por el Licenciado Elena Forcen Vazquez en la Facultad de Ciencias de la Universidad de ZaragozaResumen del trabajo presentado a la IV Reunion de Jovenes Investigadores en Coloides e Interfases, celebrada en Cordoba del 7 al 9 de febrero de 2018.Poster presentado en el II Simposio del Grupo de Trabajo “Interacciones Planta‐Suelo” Universidad de Jaen, Palacio de Najera (Antequera) 22‐25 marzo (2018)Trabajo presentado en el XIII Congreso Nacional de Acuicultura, celebrado en Barcelona del 21 al 24 de noviembre de 2011.Trabajo presentado al 15th JCF Fruhjahrssymposium (Congreso de Jovenes Investigadores de la Sociedad Alemana de Quimica) celebrado en Berlin (Alemania) del 6 al 9 de marzo de 2013.Trabajo presentado en la V Reunion de Internacional de FUEGORED (Red Tematica efectos de los incendios forestales sobre los suelos), celebrada en Barcelona – Solsona (Espana), del 8 al 10 de mayo de 2014Poster presentado a la 20th International Conference on Solid Compounds of Transition Elements, celebrada del 11 al 15 de abril de 2016 en Zaragoza (Espana).10 pages, 3 figures, 4 tables, 61 references. We thank the Consejeri´a de Medio Ambiente, Junta de Andaluci´a, for permitting the field work in Sierra Nevada and Sierra de Baza. Sergio de Haro provided invaluable help in the field. Corinna Riginos made constructive comments on the manuscript. David Nesbitt improved the English version.Trabajo presentado a la Annual World Conference on Carbon, celebrada en Dresden (Alemania) del 12 al 17 de julio de 2015.

Dose-titration of virulent Neospora caninum isolate Nc-Spain7 in pregnant sheep at 90 days of gestation

La enfermedad de Lafora es un tipo de epilepsia mioclonica progresiva de herencia autosomica recesiva causada por la mutacion de uno de los siguientes genes: EPM2A, que codifica para la fosfatasa de especificidad dual laforina, y EPM2B, que codifica para la E3 ubicuitina ligasa malina. Ambas proteinas participan en las mismas rutas fisiologicas a traves de la formacion de un complejo funcional en el que laforina recluta de manera especifica sustratos para que sean ubicuitinados por malina. Sin embargo, muchos aspectos relativos al funcionamiento de este complejo estan aun por definir. En el presente trabajo, hemos analizado el mecanismo de dimerizacion de laforina, descubriendo que la cisteina 329 juega un papel fundamental en el proceso de dimerizacion, ya que su mutacion da lugar a una forma exclusivamente monomerica que mantiene la capacidad catalitica y la capacidad de formar un complejo funcional con malina. Por otro lado, hemos definido que el complejo laforina-malina emplea a la enzima conjugadora de ubicuitina UBE2N para promover la ubicuitinacion de sus sustratos con cadenas unidas en K63. El adaptador de la autofagia selectiva p62 tambien interacciona con el complejo laforina-malina, estimulando su capacidad de ubicuitinacion y siendo ubicuitinado el mismo en el proceso. Todos ellos colocalizan en regiones donde esta teniendo lugar la autofagia. Ademas, el complejo laforina-malina participa en la regulacion de este proceso uniendo al nucleo central beclina1-Vps34-Vps15, promoviendo su ubicuitinacion, asi como la de reguladores posteriores como Atg14L y UVRAG. El complejo laforina-malina esta regulado por componentes de la maquinaria de autofagia, como Bcl-2, que inhibe su capacidad de ubicuitinacion, o TRAF6, que promueve la ubicuitinacion de laforina. Sin embargo, el complejo laforina-malina no se recluta a la mitocondria para su degradacion por autofagia ni en condiciones normales ni tras producirse dano mitocondrial. Del mismo modo, la migracion de parkina a la mitocondria para inducir su degradacion no esta afectada ni en ausencia ni en sobreexpresion de laforina y malina. En conjunto, hemos definido algunos aspectos relevantes respecto al mecanismo por el que actuan laforina y malina, y por tanto mejorado nuestra comprension sobre como pueda estar causada la patologia.290 Pags.- Figs.- Tabls. Tesis Univ. Zaragoza, Departamento de Geografia y Ordenacion del Territorio. Dirigida por los Drs. Santiago Begueria (EEAD-CSIC) y Sergio M. Vicente-Serrano (IPE-CSIC). Bajo Creative Commons License Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0).We define a clause tableau calculus for MaxSAT, prove its soundness and completeness, and describe a tableau-based algorithm for MaxSAT. Given a multiset of clauses ϕ, the algorithm computes both the minimum number of clauses that can be falsified in ϕ, and an optimal assignment. We also describe how the algorithm can be extended to solve weighted MaxSAT and weighted partial MaxSAT.Grupo 5: Valor social.-- Programa de investigacion para la conservacion preventiva y regimen de acceso de la cueva de Altamira (2012-2014).Automated planning has proven to be useful to solve problems where an agent has to maximize a reward function by executing actions. As planners have been improved to salve more expressive and difficult problems, there is an increasing interest in using planning to improve efficiency in robotic tasks. However, planners rely on a domain model, which has to be either handcrafted or learned. Although learning domain models can be very costly, recent approaches provide generalization capabilities and integrate human feedback to reduce the amount of experiences required to learn. In this thesis we propase new methods that allow an agent with no previous knowledge to solve certain problems more efficiently by using task planning. First, we show how to apply probabilistic planning to improve robot performance in manipulation tasks (such as cleaning the dirt or clearing the tableware on a table). Planners obtain sequences of actions that get the best result in the long term, beating reactive strategies. Second, we introduce new reinforcement learning algorithms where the agent can actively request demonstrations from a teacher to learn new actions and speed up the learning process. In particular, we propase an algorithm that allows the user to set the minimum quality to be achieved, where a better quality also implies that a larger number of demonstrations will be requested . Moreover, the learned model is analyzed to extract the unlearned or problematic parts of the model. This information allow the agent to provide guidance to the teacher when a demonstration is requested, and to avoid irrecoverable errors. Finally, a new domain model learner is introduced that, in addition to relational probabilistic action models, can also learn exogenous effects. This learner can be integrated with existing planners and reinforcement learning algorithms to salve a wide range of problems. In summary, we improve the use of learning and task planning to salve unknown tasks. The improvements allow an agent to obtain a larger benefit from planners, learn faster, balance the number of action executions and teacher demonstrations, avoid irrecoverable errors, interact with a teacher to solve difficult problems, and adapt to the behavior of other agents by learning their dynamics. All the proposed methods were compared with state-of-the-art approaches, and were also demonstrated in different scenarios, including challenging robotic tasks.L’estudi del sistema d’alimentacio d’hidrogen es un dels camps de recerca necessaris per a allargar la vida util de les piles de combustible PEM. Estudis anteriors han demostrat que la vida util i l’eficiencia d’aquestes piles milloren amb l’us d’un sistema de recirculacio d’hidrogen. En aquest treball s’ha dissenyat i fabricat un ejector per a ser implementat a l’estacio de proves 4 del laboratori de control de piles de combustible de l’IRI amb la finalitat d’analitzar l’efecte que els sistemes de recirculacio d’hidrogen basats en ejectors tenen sobre aquestes piles. En el disseny de la geometria d’un ejector s’han de considerar diferents parametres que nomes es poden estudiar mitjancant CFD (Computational Fluid Dynamics). Per tant, s’ha desenvolupat un model 2D axisimetric utilitzant el model de turbulencia k-e estandard per a resoldre les equacions de Navier-Stokes amb una mitjana de Favre. El model s’ha utilitzat per a dissenyar i fabricar un ejector experimental amb la finalitat de validar-lo. Els resultats obtinguts mostren que el model es capac d’obtenir els fluxos massics que apareixen en un ejector per a diferents condicions operatives. Un cop validat el model, s’ha fet servir per a fer un estudi parametric per a trobar els parametres geometrics optims que ha de tenir l’ejector dissenyat per a l’estacio de proves 4.Trabajo presentado en el 10th International Symposium on the Cretaceous, celebrado en Viena (Austria) del 21 al 26 de agosto de 2017Este trabajo ha sido parcialmente financiado por el proyecto CGL2009-11316 y por una beca de Formacion de Profesorado Universitario del Ministerio de educacion AP2007-03633.Trabajo presentado en el XXIV Congreso Iberoamericano de Catalisis, celebrado en Medellin (Colombia) del 14 al 19 de septiembre de 2014.Trabajo presentado en el 33rd International Meeting of Sedimentology y 16eme Congres Francais de Sedimentologie (2017), celebrado en Toulouse (Francia), del 10 al 12 de octubre de 2017Trabajo presentado en la XXV Reunion Bienal de Quimica Organica celebrada en Alicante del 4 al 6 de junio de 2014.Resumen del trabajo presentado a las II Jornadas Cientificas CIAL-Forum, celebradas durante los dias 16 y 17 de noviembre de 2016 en el Instituto de Investigacion en Ciencias de la Alimentacion (CIAL).Esta reflexion analiza el paralelismo entre el origen del discurso patrimonial autorizado y la precarizacion del trabajo femenino. Ambos surgen con el capitalismo y son dos de sus pilares. Plantea que el cambio simbolico que implica la igualdad legal entre hombres y mujeres supone un cambio profundo en las estructuras sociales y que este cambio ofrece serias resistencias, que se generan no solo por la necesidad de las instancias de poder de seguir ocupandolo sino porque implican cambios en la identidad de los grupos y de los individuos, con la angustia que ello genera. La idea de patrimonio cultural esta directamente implicada en estos cambios puesto que la incorporacion de las mujeres a el implica una variacion sustancial en el propio concepto. Sugiere que para avanzar en este cambio es necesario comprender que el “que” y el “como” se hacen las cosas estan necesariamente imbricados.Presentacion para el 50th European Marine Biology Symposium, 21-25 September 2015, Helgoland, Germany.-- 15 pagesResumen del trabajo presentado al XVIII Scientific Meeting of the Spanish Society of Chromatography and Related Techniques (SECyTA), celebrado en Granada del 2 al 4 de octubre de 2018.Resumen del trabajo presentado al 42st Annual Association for Research in Otolaryngology (ARO) MidWinter Meeting, celebrado en Baltimore, Maryland (USA) del 9 al 13 de febrero de 2019.Partial financial support has been received from the Agencia Estatal de Investigacion (AEI, MCI, Spain) and Fondo Europeo de Desarrollo Regional (FEDER, UE), under Project PACSS (RTI2018-093732-B-C21/C22) and the Maria de Maeztu Program for units of Excellence in R&D (MDM-2017-0711). A.F.P. acknowledges support by the Formacion de Profesorado Universitario (FPU14/00554) program of Ministerio de Educacion, Cultura y Deportes (MECD) (Spain).Memoria presentada para optar al titulo de Doctor otorgado por la Universidad de Cantabria por Biuse Casaponsa Gali y que ha sido realizada en el Instituto de Fisica de Cantabria.Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (Madrid, Spain. 11-14 October 2017).3 paginas, 1 tabla , 1 figura.--Trabajo presentado a las XVII Jornadas sobre Produccion Animal AIDA (Zaragoza, 30 y 31 de mayo, 2017).301 paginas.-- 392 referencias.-- Memoria para optar al titulo de Doctora en Farmacia por la Universidad de SevillaTrabajo presentado en el ANQUE.ICCE.BIOTEC - Congress on Chemistry, Chemical Engineering and Biotechnology, celebrado en Madrid del 01 al 04 de julio de 2014.Towards an integrated evaluation of the Spanish journals on Social and Human Sciences. The journals of Psychology. The present work is part of an ampler project about Spanish journals evaluation of Social y Human Sciences. First, the results of the evaluation of the Spanish journals of Psychology are presented. These results are obtained from a survey sent to university professors in order to evaluate the journals to establish hierarchized relations in each area of knowledge. Second, a study of the references cited from a selection of journals is realized. Finally, an integrated table of valuation of the journals is presented. In order to facilitate the task a data base is elaborated. Among the obtained conclusions it is particularly interested the fact that while the professors declare to publish mainly in Spanish journals, they preferredly cite foreign journals. On the other hand, it is indicated that the different methods to evaluate a journal are complementary and it is important to know the used method in each case.Trabajo presentado al: 4th International Meeting on Apicomplexa in Farm Animals. (11-14 October 2017 - Madrid, Spain).Trabajo presentado en la ed. anual 2017 del congreso nacional BIOTEC organizado por la Sociedad Espanola de Biotecnologia en el Campus La Merced de la Universidad de Murcia durante los dias 18 a 21 de junio de 2017.Memoria presentada para optar al grado de Doctor en Ciencias, Seccion Quimicas, por el Licenciado Elena Forcen Vazquez en la Facultad de Ciencias de la Universidad de ZaragozaResumen del trabajo presentado a la IV Reunion de Jovenes Investigadores en Coloides e Interfases, celebrada en Cordoba del 7 al 9 de febrero de 2018.Poster presentado en el II Simposio del Grupo de Trabajo “Interacciones Planta‐Suelo” Universidad de Jaen, Palacio de Najera (Antequera) 22‐25 marzo (2018)Trabajo presentado en el XIII Congreso Nacional de Acuicultura, celebrado en Barcelona del 21 al 24 de noviembre de 2011.Trabajo presentado al 15th JCF Fruhjahrssymposium (Congreso de Jovenes Investigadores de la Sociedad Alemana de Quimica) celebrado en Berlin (Alemania) del 6 al 9 de marzo de 2013.Trabajo presentado en la V Reunion de Internacional de FUEGORED (Red Tematica efectos de los incendios forestales sobre los suelos), celebrada en Barcelona – Solsona (Espana), del 8 al 10 de mayo de 2014Poster presentado a la 20th International Conference on Solid Compounds of Transition Elements, celebrada del 11 al 15 de abril de 2016 en Zaragoza (Espana).10 pages, 3 figures, 4 tables, 61 references. We thank the Consejeri´a de Medio Ambiente, Junta de Andaluci´a, for permitting the field work in Sierra Nevada and Sierra de Baza. Sergio de Haro provided invaluable help in the field. Corinna Riginos made constructive comments on the manuscript. David Nesbitt improved the English version.Trabajo presentado a la Annual World Conference on Carbon, celebrada en Dresden (Alemania) del 12 al 17 de julio de 2015.