Síntese de tetrahidroquinolinas a partir da D-eritrose

Dissertação de mestrado em Química Medicinal

Síntese de novas tieno[3,2-b]piridinas com potencial atividade biológica contendo 1,2,3-triazoles 1,4-dissubstituídos obtidos por reação "click" de ciclioadição azida-alquino catalisada por Cu(I)-CuAAC

Dissertação de mestrado em Química Medicinal

The role of monocarboxylate transporters in 3-bromopyruvate effect in cancer cells

Tese de Doutoramento em Biologia Molecular e Ambiental (área de especialização em Biologia Molecular e Saúde).

Participación del tráfico y la distribución de proteínas en la polaridad y el desarrollo de procesos neuronales. Protein sorting and trafficking participation in neuronal processes polarity and development.

El estudio del tráfico intracelular en neuronas ha despertado gran interés en los últimos años, debido a que un gran número de enfermedades neurodegenerativas y neuropsiquiátricas parecen tener origen en en el transporte defectuoso de proteínas en estos tipos celulares. Mediante el uso de técnicas de biología celular y molecular, fuimos capaces de describir una de las vías que regula la fisión de las vesículas que llevan su cargo desde la última cisterna del Aparato de Golgi hacia la superficie celular en células epiteliales no polarizadas. Uno de los componentes clave de esa vía resultó ser la Proteina Kinasa D1 (PKD1), cuya actividad en el Aparato de Golgi es esencial para un normal transporte intracelular. Sorprendentemente, observamos que la PKD1 en neuronas con polaridad establecida no regula la fisión en el Golgi, pero si estaría involucrada en la selectividad y distribución (sorting) de vesículas cuyo cargo debe ser específicamente dirigido a las membranas dendríticas. El bloqueo de la actividad de la PKD1 no solamente cambia el destino final de estos cargos, que son enviados de esta forma a la membrana terminal del axón, sino que también es capaz de inducir defectos en el desarrollo y crecimiento de los procesos dendríticos a largo plazo. En este proyecto estudiaremos de que manera influye la perturbación del sorting, en ausencia de PKD1 activa y de otros componentes que la regulan, en la distribución de receptores de factores neurotróficos y de neurotransmisores glutamatérgicos, y cómo estos cambios en su distribución afectan el número, tamaño, y funcionalidad de los procesos neuronales (axones y dendritas). Estos resultados contribuirán a adquirir mayores conocimientos de los mecanismos dependientes del transporte y sorting de proteínas de membrana que participan en la regulación del crecimiento neuronal, los cuales a su vez aportarán información valiosa en la comprensión de un gran número de enfermedades neurológicas. The study of intracellular trafficking in neurons has arisen a great deal of interest in the last years, since a great number of neurodegenerative and neuropsychiatric disorders seem to be originated in abnormal protein transport in these type of cells. Using cell and molecular biology methodologies, we have been capable of describe one of the pathways that regulate the fission of vesicles that carry their cargo from the last Golgi Apparatus cisternae to the cell surface in non-polarized epithelial cells. One of the key components in this pathway is the Protein Kinase D1 (PKD1), whose activity in the Golgi Apparatus is essential for a normal intracelular transport. Surprisingly, we have observed that PKD1 does not regulate fission in neurons with established polarity, but it would be involved in vesicles' sorting at Golgi, particularly of those that carry specific dendritic cargo. Blocking PKD1 activity changes the final destination of these cargoes, which is now sent to the axons' terminal membranes, and also produces late dendritic development and growing defects. In this project we will study how sorting perturbation in absence of PKD1 and its regulators activities influences selectivity and distribution of neurotrophic and neurotransmitter receptors, and how these sorting changes affect number, size and functionality of neuronal processes (axons and dendrites). These results will help to acquire greater knowledge about transport and sorting mechanisms of neuronal growth regulatory membrane proteins. In addition, these studies will contribute with new valuable information necessary to understand numerous neurological diseases.

Age, growth and population dynamics of lemon sole Microstomus kitt (Walbaum 1792) sampled off the West Coast of Ireland

The age, growth, maturity and population dynamics of lemon sole (Microstomus kitt), captured off the west coast of Ireland (ICES division Vllb), were determined for the period November 2000 to February 2002. The maximum age recorded was 14 years. Males of the population were dominated by 4 year olds, while females were dominated by 5 year olds. Females dominated the sex ratio in the overall sample, each month sampled, at each age and from 22cm in total length onwards (when N > 20). Possible reasons for the dominance of females in the sex ratio are discussed. Three models were used to obtain the parameters of the von Bertalanfly growth equation. These were the Ford-Walford plot (Beverton and Holt 1957), the Gulland and Holt plot (1959) and the Rafail (1973) method. Results of the fitted von Bertalanffy growth curves showed that female lemon sole o f f the west coast of Ireland grew faster than males and attained a greater size. Male and female lemon sole mature from 2 years of age onwards. There is evidence in the population o f a smaller asymptotic length (L«, = 34.47cm), faster growth rate (K = 0.1955) and younger age at first maturity, all of which are indicative o f a decrease in population size, when present results are compared to data collected in the same area 22 years earlier. Results of the yield per recruit curve indicate that lemon sole are currently being over-fished o f f the west coast of Ireland. Problems of selectivity within the sampling method, particularly at the discarding stage, may have influenced the outcome of results of the models used in the assessment of this stock. Therefore, additional/future work on this species should include catch data which incorporates discards and not landings data alone.

Einfluss der Mikrokonvektion auf Selektivität und Stofftransport während der Verdunstung binärer Flüssigkeiten aus porösen Schichten

Interfacial convection, enhancement of mass transfer, packed beds, evaporation, selectivity

Flow and transport in electrochromatography

Analyte retention, analyte transport, axial dispersion, adsorption, charge-selectivity, concentration polarization, confocal laser scanning microscopy, donnan-exclusion, electrical double layer; electrochromatography; electrohydrodynamics, electrokinetic instability, electroosmosis; electroosmotic flow; electroosmotic mobility, electroosmotic perfusion, electrophoresis, hierarchical porous media, hydrodynamic flow, induced-charge electroosmosis, ion-permselectivity, ion-permselective transport, monolith, nonequilibrium electrical double layer, nonequilibrium electrokinetic effects, nonlinear electroosmosis, plate height, plate number, porous media, pore-scale dispersion, refractive index matching, space charge effects, sphere packing, quantitative imaging, wall effect, zeta-potential

Fingermarks and Other Impressions Left by the Human Body - A Review (August 2007-July 2010)

The purpose of this paper is to review the scientific literature from August 2007 to July 2010. The review is focused on more than 420 published papers. The review will not cover information coming from international meetings available only in abstract form. Fingermarks constitute an important chapter with coverage of the identification process as well as detection techniques on various surfaces. We note that the research has been very dense both at exploring and understanding current detection methods as well as bringing groundbreaking techniques to increase the number of marks detected from various objects. The recent report from the US National Research Council (NRC) is a milestone that has promoted a critical discussion on the state of forensic science and its associated research. We can expect a surge of interest in research in relation to cognitive aspect of mark and print comparison, establishment of relevant forensic error rates and statistical modelling of the selectivity of marks' attributes. Other biometric means of forensic identification such as footmarks or earmarks are also covered in the report. Compared to previous years, we noted a decrease in the number of submission in these areas. No doubt that the NRC report has set the seed for further investigation of these fields as well.

How Effective are Unemployment Benefit Sanctions? Looking Beyond Unemployment Exit

This paper provides a comprehensive evaluation of the effects of benefit sanctions on post-unemployment outcomes such as post-unemployment employment stability and earnings. We use rich register data which allow us to distinguish between a warning that a benefit reduction may take place in the near future and the actual withdrawal of unemployment benefits. Adopting a multivariate mixed proportional hazard approach to address selectivity, we find that warnings do not affect subsequent employment stability but do reduce post-unemployment earnings. Actual benefit reductions lower the quality of post-unemployment jobs both in terms of job duration as well as in terms of earnings. Copyright © 2012 John Wiley & Sons, Ltd.

Ultra high performance supercritical fluid chromatography coupled with tandem mass spectrometry for screening of doping agents. II: Analysis of biological samples.

The potential and applicability of UHPSFC-MS/MS for anti-doping screening in urine samples were tested for the first time. For this purpose, a group of 110 doping agents with diverse physicochemical properties was analyzed using two separation techniques, namely UHPLC-MS/MS and UHPSFC-MS/MS in both ESI+ and ESI- modes. The two approaches were compared in terms of selectivity, sensitivity, linearity and matrix effects. As expected, very diverse retentions and selectivities were obtained in UHPLC and UHPSFC, proving a good complementarity of these analytical strategies. In both conditions, acceptable peak shapes and MS detection capabilities were obtained within 7min analysis time, enabling the application of these two methods for screening purposes. Method sensitivity was found comparable for 46% of tested compounds, while higher sensitivity was observed for 21% of tested compounds in UHPLC-MS/MS and for 32% in UHPSFC-MS/MS. The latter demonstrated a lower susceptibility to matrix effects, which were mostly observed as signal suppression. In the case of UHPLC-MS/MS, more serious matrix effects were observed, leading typically to signal enhancement and the matrix effect was also concentration dependent, i.e., more significant matrix effects occurred at the lowest concentrations.

Labor supply response to international migration and remittances in the Republic of Haiti

The Republic of Haiti is the prime international remittances recipient country in the Latin American and Caribbean (LAC) region relative to its gross domestic product (GDP). The downside of this observation may be that this country is also the first exporter of skilled workers in the world by population size. The present research uses a zero-altered negative binomial (with logit inflation) to model households' international migration decision process, and endogenous regressors' Amemiya Generalized Least Squares method (instrumental variable Tobit, IV-Tobit) to account for selectivity and endogeneity issues in assessing the impact of remittances on labor market outcomes. Results are in line with what has been found so far in this literature in terms of a decline of labor supply in the presence of remittances. However, the impact of international remittances does not seem to be important in determining recipient households' labor participation behavior, particularly for women.

Estudi de la reductibilitat de catalitzadors basats en cobalt i el seu comportament en la reacció de desplaçament de gas d'aigua de CO

Projecte de recerca elaborat a partir d’una estada a la universitat d'Udine, Itàlia, entre setembre i desembre del 2006.S'han caracteritzat mitjançant la reducció a temperatura programada i tests catalítics catalitzadors en pols basats en cobalt i supostats en òxid de zinc i monòlits ceràmics funcionaliltzats també amb cobalt i òxid de zinc. L'addició de promotors (manganès, crom i ferro ) als catalitzadors en pols, preparats per impregnació i precipitació, no afecta significativament ni la temperatura a la qual té lloc la reducció ni al percentatge global de reducció. En els cicles de reducció-oxidació sí que s'observen diferències entre el primer perfil de reducció i els següents, especialment en el cas de la mostra que té ferro com a promotor, on les diferències s'accentuen en cicles successius (fins al quart). S'ha evaluat l'activitat d'aquests catalitzadors en la reacció de desplaçament de gas d'aigua, obtenint uns resultats satisfactoris. Finalment s'han realitzat reduccions a temperatura programada i tests catalítics en la reacció de desplaçament de gas d'aigua amb monòlits funcionalitzats amb cobalt i òxid de zinc (en cap d'ells s'ha introduït promotors). El nivell de conversió assolit és menor que en el cas de catalitzadors en pols, fet que s'associa a la geometria d'aquests sistemes catalítics, però la relació CH4/CO2 és més favorable que en els catalitzadors en pols, el que els converteix en sistemes molt selectius.

Entrepreneurial Orientation, Intangible Assets and Firm Growth: the impact of ‘Spirit and Material’ on the growth of Chinese private firms

This paper has three contributions. First, it shows how field work within small firms in PR Chinese has provided new evidence which enables us to measure and calibrate Entrepreneurial Orientation (EO), as ‘spirit’, and Intangible Assets (IA), as ‘material’, for use in models of small firm growth. Second, it uses inter-item correlation analysis and both exploratory and confirmatory factor analysis to provide new measures of EO and IA, in index and in vector form, for use in econometric models of firm growth. Third, it estimates two new econometric models of small firm employment growth in PR China, under the null hypothesis of Gibrat’s Law, using our two new index-based and vector-based measures of EO and IA. Estimation is by OLS with adjustment for heteroscedasticity, and for sample selectivity. Broadly, it finds that EO attributes have had little significant impact on small firm growth, and indeed innovativeness and pro-activity paradoxically may even dampen growth. However, IA attributes have had a positive and significant impact on growth, with networking, and technological knowledge being of prime importance, and intellectual property and human capital being of lesser but still significant importance. In the light of these results, Gibrat’s Law is generalized, and Jovanovic’s learning theory is extended, to emphasise the importance of IA to growth. These findings cast new empirical light on the oft-quoted national slogan in PR China of “spirit and material”. So far as small firms are concerned, this paper suggests that their contribution to PR China’s remarkable economic growth is not so much attributable to the ‘spirit’ of enterprise (as suggested by propaganda) as, more prosaically, to the pursuit of the ‘material’.

Combinatorial Synthesis and Biological Evaluation of Inhibitors of D-Ala-D-Ala-Ligase

Report for the scientific sojourn carried out at the Max Planck Institut of Molecular Phisiology, Germany, from 2006 to 2008.The work carried out during this postdoctoral stage was focused on two different projects. Firstly, identification of D-Ala D-Ala Inhibitors and the development of new synthethic approaches to obtain lipidated peptides and proteins and the use of these lipidated proteins in biological and biophysical studies. In the first project, new D-Ala D-Ala inhibitors were identified by using structural alignments of the ATP binding sites of the bacterial ligase DDl and protein and lipid kinases in complex with ATP analogs. We tested a series of commercially available kinase inhibitors and found LFM-A13 and Tyrphostine derivatives to inhibit DDl enzyme activity. Based on the initial screening results we synthesized a series of malononitrilamide and salicylamide derivatives and were able to confirm the validity of these scaffolds as inhibitors of DDl. From this investigation we gained a better understanding of the structural requirements and limitations necessary for the preparation of ATP competitive DDl inhibitors. The compounds in this study may serve as starting points for the development of bi-substrate inhibitors that incorporate both, an ATP competitive and a substrate competitive moiety. Bisubstrate inhibitors that block the ATP and D-Ala binding sites should exhibit enhanced selectivity and potency profiles by preferentially inhibiting DDl over kinases. In the second project, an optimized synthesis for tha alkylation of cysteins using the thiol ene reaction was establisehd. This new protocol allowed us to obtain large amounts of hexadecylated cysteine that was required for the synthesis of differently lipidated peptides. Afterwards the synthesis of various N-ras peptides bearing different lipid anchors was performed and the peptides were ligated to a truncated N-ras protein. The influence of this differently lipidated N-ras proteins on the partioning and association of N-Ras in model membrane subdomains was studied using Atomic Force Microscopy.

Preparació, estudi estructural i aplicabilitat de derivats de Poli-(4R)-hidroxi-L-prolina a la separació d’enantiòmers per tècniques cromatogràfiques o relacionades

La separació d’enantiòmers (isòmers òptics) és molt important en molts diversos camps, com les síntesis quirals, biologia, i en el camp de la farmacologia especialment. És per això, que es fa necessari de disposar de tècniques i mètodes analítics ràpids, fiables i sensibles per a la separació d’enantiòmers. La present tesi s’emmarca en el camp de la separació d’enantiòmers, concretament en la preparació de fases estacionàries quirals per al seu ús en cromatografia liquida. En aquest sentit, s’ha desenvolupat la síntesi i caracterització de molècules polimèriques quirals derivades de l’aminoàcid L-prolina que incorporades en matrius de gel de sílice poden constituïr columnes quirals per a la separació d’enantiòmers per cromatografia liquida. S’han estudiat les característiques enantioselectives d’aquests nous materials en la separació de molècules quirals, trobant-se ésser satisfactòriament enantioselectius. L’interès que suscita l’obtenció d’enantiòmers a gran escala fa que la recerca s’orienti a la recerca de materials amb elevada capacitat de càrrega, és a dir, que puguin donar lloc a la separació d’elevades quantitats d’enantiòmers. Amb aquesta finalitat s’han dut a terme assaigs de capacitat de càrrega, que han posat de manifest la possible aplicació d’aquests materials a la separació preparativa d’enantiòmers. També s’ha parat especial atenció a l’estudi de les característiques de la matriu de gel de sílice, assajant-se altres materials de sílice més porosos i que permeten així treballar amb fluxos més elevats tot reduint-ne el temps d’anàlisi i els costos associats a la separació preparativa d’enantiòmers. L'estudi conformacional d'aquests nous selectors també ha estat contemplat per tal d'explicar l'enantioselectivitat específica que s'observa en certs dissolvents orgànics en els qual es duu a terme la separació dels enantiòmers.