Rafael Rodríguez Martínez

Immunological evaluation in nonhuman primates of formulations based on the chimeric protein P64k-domain III of dengue 2 and two components of Neisseria meningitidis.

The main problem in the development of successful vaccines against dengue based on recombinant proteins is the necessity to use potent adjuvants to reach a proper functional immune response. Our group reported the expression, characterization and immunological evaluation of the recombinant protein PD5, which contains the domain III of the Envelope protein from dengue 2 virus fused to the carrier protein P64k. This construct completely protected monkeys against viral challenge when the Freunds adjuvant was employed. Therefore, to define suitable formulations for human use, the present work relies on the evaluation of PD5, produced with a high purity and under GMP conditions, when formulated either with outer membrane vesicles (OMV) or the serogroup A capsular polysaccharide (CPS-A) from Neisseria meningitidis, both adsorbed on aluminium hydroxide. The antibody response to the formulation containing the CPS-A was clearly superior to that of the formulation with OMV. The experiment of in vivo protection supported this evidence, since only the group immunized with PD5 and CPS-A was partially protected upon viral challenge. This is the first study in which the polysaccharide A of N. meningitidis is successfully employed as adjuvant for viral antigens.

Primary and Secondary Infections of Macaca fascicularis Monkeys with Asian and American Genotypes of Dengue Virus 2

ABSTRACT The goal of this study was to compare the immune response and the protection capacity induced by the dengue virus 2 (DENV-2) American and Asian genotypes in Macaca fascicularis monkeys. Animals were infected with American or Asian DENV-2 strains and challenged 1 year later with a DENV-2 Asian genotype strain. The viremia and monkey antibody levels were similar for the different strains after primary and secondary infection; however, the functionality of the antibody response was different. A limited viral replication was demonstrated after the secondary infection in all the monkeys. No virus was isolated in tissue culture, while reverse transcription-PCR showed a late positive reaction in four of five challenged monkeys. The immunoglobulin M response pattern and the detection of antibodies to specific proteins by Western blotting supported the protection data. Despite the demonstration of the protective effect after homologous challenge, a strong anamnestic antibody response was observed.

Synthesis and highly selective Diels–Alder cycloadditions of the new dienes N-substituted 2,3,5,6-tetrahydrobenzoxazol-2-ones

Abstract The synthesis of the new dienes N -substituted 2,3,4,5-tetrahydrobenzoxazol-2-ones 8a – 8c is described, through a one-step convergent process from 1,2-cyclohexanedione ( 7a ) and the corresponding isocyanates 2a – 2c . The presence of electron-donor substituents in the aryl ring of the isocyanate gave rise to the exclusive formation of the captodative olefins 10 . Diene 8a proved to be reactive and stereoselective in Diels–Alder additions with a cyclic olefin. The reaction with acetylenic dienophiles yielded the 2,3-dihydrobenzoxazol-2-ones 21 and 24 , as the products of sequential [4+2] cycloaddition and retro-Diels–Alder reactions. Methyl vinyl ketone ( 22 ) underwent regio- and stereoselective tandem Diels–Alder and Michael additions to give propellane 29a . Evidence of an endo π-pyramidalization of the central double bond of adduct 19 would rationalize the exo stereoselection in the formation of 29a . The regioselectivity in these reactions has been rationalized in terms of FMO theory by ab initio calculations.

Regioselective Synthesis of N-Substituted 4-Methylene-2-oxazolidinones and 4-Oxazolin-2-ones. Study of Reactivity in Thermal Michael Conjugate Additions

N-Substituted 4-methylene-2-isoxazolidinones 9a–9e have been prepared from the tandem condensation of isocyanates 3 with α-ketol 7. In a more polar solvent (DMF) the same reaction led to stereoisomeric alcohols 11 and 12, which could be transformed to the thermodynamically more stable isomers 4-oxazolin-2-ones 10 in good yield. Thermal conjugate additions of both heterocycles 9a and 10a to enone 13 provided the C-5 adduct 15. When the reaction was carried out with the captodative olefin 17, the unexpected aniline 18 was isolated. The regiochemistry of the Michael addition was rationalized in terms of FMO theory by ab initio calculations.

Stereoselective Synthesis of 4,5-Diethylidene-Oxazolidinones as New Dienes in Diels-Alder Reactions

The N-substituted isomeric (4Z,5Z)- and (4E,5Z)-4,5-diethylideneoxazolidin-2-ones 5 and 6 were synthesized, the latter being favored during the one-step process from the α-diketone 1c and different isocyanates. The steric interaction between the N-substituent and the Me group attached to the exocyclic diene moiety plays a decisive role in controlling the observed stereoselectivity, as suggested by the calculated free energies of the two isomers. Both dienes undergo efficient additions to symmetric dienophiles in thermal Diels-Alder reactions to yield the adducts 11 and 13, respectively. These molecules displayed interesting C−H⋅⋅⋅π, and C−H⋅⋅⋅X (X=O, Cl) interactions according to their X-ray crystal structures. Isomers 6 suffered highly stereo- and regioselective additions with nonsymmetrical dienophiles such as methyl vinyl ketone or methyl propiolate. Steric interactions, promoted by the inward-pointing Me group in 6, seem to explain such selectivity. These results have also been rationalized by ab initio calculations in terms of the FMO theory.

Design and Manufacture of a Prototype of a Testing Rig for the Evaluation of the Biomechanical Behavior of Vertebrae

The objective of this work is the conceptual design of a testing rig for the biomechanical evaluation of vertebral bodies. Such a rig must test diverse vertebral specimens under compression, flexion and the combination of both loading cases. The dimensions of the porcine and human vertebrae were taken into account. Special attention was given to the design of the grips, in such a way that the range of movements of the spine is reproduced. The structural integrity of this rig was determined by means of the finite element method. In a following step, compression, bending and compression and bending tests of a C3–C7 vertebral body were simulated. This data was obtained from an individual, who was 55 years old. In all these cases, the testing rig had stresses which were within the allowable range. The resultant stresses on the vertebral bodies were in line with those reported in the open literature. In the next step, a “quick” prototype was manufactured in a 3D printer (Dimension SST 1,200). Finally, a data acquisition system has been proposed.

Más allá de la arqueología pública: arqueología, democracia y comunidad en el yacimiento multivocal de A Lanzada (Sanxenxo, Pontevedra)

La Diputacio Provincial de Pontevedra i l’Institut de Ciencies del Patrimoni del CSIC estan desenvolupant des del maig del 2010 un projecte de posada en valor de l’area arqueologica d’A Lanzada (Sanxenxo, Pontevedra) a la zona zero del turisme estival a les Rias Baixas. Aquest projecte de gestio integral del patrimoni compta amb tres linies especifiques d’investigacio aplicada en difusio, comunicacio i didactica amb les quals busquem no sols la interaccio amb la comunitat local (propietaria del jaciment), sino tambe la seva participacio activa en el desenvolupament dels treballs. A Lanzada es un espai social viu, amb un profund rerefons simbolic per als habitants de la zona, una area multidimensional en la qual conflueixen diferents discursos i experiencies. Aquesta circumstancia ha de ser tinguda en compte per una arqueologia publica que ha d’anar mes enlla de la transparencia informativa i la divulgacio per materialitzar processos d’apoderament propis d’una arqueologia en comunitat . En la nostra comunicacio exposarem l’estrategia de treball adoptada i les activitats (encara en curs) realitzades amb les associacions locals, des de visites guiades, participacio activa en les excavacions, historia oral, etnoarqueologia, recuperacio de la memoria historica..., fins a exposicions interactives i cicles de conferencies i debat a la parroquia de Noalla. Al seu torn, contextualitzarem el projecte en el marc de la linia de treball en cultura cientifica desenvolupada en els ultims anys per l’INCIPT a Galicia.

Beyond public archaeology: archeology, democracy and community in the multivocal site of A Lanzada (Sanxenxo, Pontevedra)

La Diputacio Provincial de Pontevedra i l’Institut de Ciencies del Patrimoni del CSIC estan desenvolupant des del maig del 2010 un projecte de posada en valor de l’area arqueologica d’A Lanzada (Sanxenxo, Pontevedra) a la zona zero del turisme estival a les Rias Baixas. Aquest projecte de gestio integral del patrimoni compta amb tres linies especifiques d’investigacio aplicada en difusio, comunicacio i didactica amb les quals busquem no sols la interaccio amb la comunitat local (propietaria del jaciment), sino tambe la seva participacio activa en el desenvolupament dels treballs. A Lanzada es un espai social viu, amb un profund rerefons simbolic per als habitants de la zona, una area multidimensional en la qual conflueixen diferents discursos i experiencies. Aquesta circumstancia ha de ser tinguda en compte per una arqueologia publica que ha d’anar mes enlla de la transparencia informativa i la divulgacio per materialitzar processos d’apoderament propis d’una arqueologia en comunitat . En la nostra comunicacio exposarem l’estrategia de treball adoptada i les activitats (encara en curs) realitzades amb les associacions locals, des de visites guiades, participacio activa en les excavacions, historia oral, etnoarqueologia, recuperacio de la memoria historica..., fins a exposicions interactives i cicles de conferencies i debat a la parroquia de Noalla. Al seu torn, contextualitzarem el projecte en el marc de la linia de treball en cultura cientifica desenvolupada en els ultims anys per l’INCIPT a Galicia.

Més enllà de l’arqueologia pública: arqueologia, democràcia i comunitat en el jaciment multivocal d’A Lanzada (Sanxenxo, Pontevedra)

La Diputacio Provincial de Pontevedra i l’Institut de Ciencies del Patrimoni del CSIC estan desenvolupant des del maig del 2010 un projecte de posada en valor de l’area arqueologica d’A Lanzada (Sanxenxo, Pontevedra) a la zona zero del turisme estival a les Rias Baixas. Aquest projecte de gestio integral del patrimoni compta amb tres linies especifiques d’investigacio aplicada en difusio, comunicacio i didactica amb les quals busquem no sols la interaccio amb la comunitat local (propietaria del jaciment), sino tambe la seva participacio activa en el desenvolupament dels treballs. A Lanzada es un espai social viu, amb un profund rerefons simbolic per als habitants de la zona, una area multidimensional en la qual conflueixen diferents discursos i experiencies. Aquesta circumstancia ha de ser tinguda en compte per una arqueologia publica que ha d’anar mes enlla de la transparencia informativa i la divulgacio per materialitzar processos d’apoderament propis d’una arqueologia en comunitat . En la nostra comunicacio exposarem l’estrategia de treball adoptada i les activitats (encara en curs) realitzades amb les associacions locals, des de visites guiades, participacio activa en les excavacions, historia oral, etnoarqueologia, recuperacio de la memoria historica..., fins a exposicions interactives i cicles de conferencies i debat a la parroquia de Noalla. Al seu torn, contextualitzarem el projecte en el marc de la linia de treball en cultura cientifica desenvolupada en els ultims anys per l’INCIPT a Galicia.