G-quadruplex ligands for cancer therapy

DNA may fold into a diversity of structures and topologies such as duplexes and triplexes. Some specific guanine-rich DNA sequences may even fold into a higher order structures denominated guanine G-quadruplexes (G4). These G-quadruplex forming sequences have shown biological interest since were found in telomeres and in promoter region of oncogenes. Thus, these G4 forming sequences have been explored as therapeutic targets for cancer therapy, since G4 formation was demonstrated to inhibit RNA-polymerase and telomerase activity. However, the G4 structures are transient and are only formed under specific conditions. Hence the main objective of this work is to develop new G4-specific ligands which may potentially find applications in the therapeutic area. Several potential G4-binding ligands were synthesized and characterized. The synthesis of these compounds consisted on a procedure based on van Leusen chemistry and a cross-coupling reaction through C-H activation, affording phenanthroline compounds (Phen-1, 50%; Phen-2, 20%), phenyl (Iso-1, 61%; Iso-2, 21%; Ter-1, 85%; Ter-2, 35%), and quinolyl (Quin-1, 85%; Quin-2, 45%) compounds. Screening assays for selecting the potential G4 compounds were performed by FRET-melting, G4-FID, CD-melting and DSF. Qualitative biophysical studies were performed by fluorescence and CD spectroscopy. Two high-specific G-quadruplex ligands, Phen-1 and Phen-2, were found to effectively bind telomeric and c-myc G4 structures. Phen-1 was found to stabilize parallel telomeric 22AG and c-myc sequence by 4.1 and 4.3 ˚C, respectively. Phen-2 also displayed high affinity towards 22AG (

Vulnerability of mediterranean watersheds to climate change

Desertification is a critical issue for Mediterranean drylands. Climate change is expected to aggravate its extension and severity by reinforcing the biophysical driving forces behind desertification processes: hydrology, vegetation cover and soil erosion. The main objective of this thesis is to assess the vulnerability of Mediterranean watersheds to climate change, by estimating impacts on desertification drivers and the watersheds’ resilience to them. To achieve this objective, a modeling framework capable of analyzing the processes linking climate and the main drivers is developed. The framework couples different models adapted to different spatial and temporal scales. A new model for the event scale is developed, the MEFIDIS model, with a focus on the particular processes governing Mediterranean watersheds. Model results are compared with desertification thresholds to estimate resilience. This methodology is applied to two contrasting study areas: the Guadiana and the Tejo, which currently present a semi-arid and humid climate. The main conclusions taken from this work can be summarized as follows: • hydrological processes show a high sensitivity to climate change, leading to a significant decrease in runoff and an increase in temporal variability; • vegetation processes appear to be less sensitive, with negative impacts for agricultural species and forests, and positive impacts for Mediterranean species; • changes to soil erosion processes appear to depend on the balance between changes to surface runoff and vegetation cover, itself governed by relationship between changes to temperature and rainfall; • as the magnitude of changes to climate increases, desertification thresholds are surpassed in a sequential way, starting with the watersheds’ ability to sustain current water demands and followed by the vegetation support capacity; • the most important thresholds appear to be a temperature increase of +3.5 to +4.5 ºC and a rainfall decrease of -10 to -20 %; • rainfall changes beyond this threshold could lead to severe water stress occurring even if current water uses are moderated, with droughts occurring in 1 out of 4 years; • temperature changes beyond this threshold could lead to a decrease in agricultural yield accompanied by an increase in soil erosion for croplands; • combined changes of temperature and rainfall beyond the thresholds could shift both systems towards a more arid state, leading to severe water stresses and significant changes to the support capacity for current agriculture and natural vegetation in both study areas.

Supporting On-Line Distributed Monitoring and Debugging

Monitoring systems have traditionally been developed with rigid objectives and functionalities, and tied to specific languages, libraries and run-time environments. There is a need for more flexible monitoring systems which can be easily adapted to distinct requirements. On-line monitoring has been considered as increasingly important for observation and control of a distributed application. In this paper we discuss monitoring interfaces and architectures which support more extensible monitoring and control services. We describe our work on the development of a distributed monitoring infrastructure, and illustrate how it eases the implementation of a complex distributed debugging architecture. We also discuss several issues concerning support for tool interoperability and illustrate how the cooperation among multiple concurrent tools can ease the task of distributed debugging.

Biochemical and spectroscopic characterization of the membrane-bound nitrate reductase from Marinobacter hydrocarbonoclasticus 617

J Biol Inorg Chem (2008) 13:1321–1333 DOI 10.1007/s00775-008-0416-1

Synthesis of precursors of the rare 3-O-methylmannose polysaccharides present in Nontuberculous Mycobacteria

3-O-methylmannose polysaccharides (MMPs) are cytoplasmic carbohydrates synthesized by mycobacteria, which play important intracellular roles, such as for example in metabolism regulation. An important way to confirm if the inhibition of the synthesis of these polysaccharides will critically affect the survival of mycobacteria is the study of the biosynthetic pathways from these molecules on these microorganisms. The purpose of this work is the efficient synthesis of three saccharides, which are rare cellular precursors from the biosynthesis of the mycobacterial polysaccharides, allowing its study. In order to obtain these molecules, a chemical strategy to connect two precursors was used. This process is called chemical glycosylation and its importance will be highlighted as an important alternative to enzymatic glycosylation. The first objective was the synthesis of the disaccharides Methyl (3-O-methyl-α-D-mannopyranosyl)-(1→4)-3-O-methyl-α-D-mannopyranoside and (3-O-Methyl-α-D-mannopyra- nosyl)-(1→4)-3-O-methyl-(α/β)-D-mannopyranose. The mannose precursors were prepared before the glycosylation reaction. The same mannosyl donor was used in the preparation of both molecules and its efficient synthesis was achieved using a 8 step synthetic route from D-mannose. A different mannosyl acceptor was used in the synthesis of each disaccharide and their syntheses were also efficient, the first one a 4 step synthetic route from α-methyl-D-mannose and the second one as an intermediate from the synthesis of the mannosyl donor. The stereoselective preparation of these disaccharides was performed successfully. The second and last objective of the proposed work was the synthesis of the tetrasaccharide methyl (3-O-methyl-α-D-mannopyranosyl-(1→4)-3-O-methyl-α-D-mannopyra- nosyl-(1→4)-3-O-methyl-α-D-mannopyranosyl-(1→4)-3-O-methyl-α-D-mannopyranoside. The disaccharide acceptor and donor to be linked through a stereoselective glycosidic reaction had to be first synthesized. Several synthetic strategies were studied. Neither the precursors nor the tetrasaccharide were synthesized, but a final promising synthetic route for its preparation has been proposed.