A new class of glycomimetic drugs to prevent free fatty acid-induced endothelial dysfunction

Background: Carbohydrates play a major role in cell signaling in many biological processes. We have developed a set of glycomimetic drugs that mimic the structure of carbohydrates and represent a novel source of therapeutics for endothelial dysfunction, a key initiating factor in cardiovascular complications. Purpose: Our objective was to determine the protective effects of small molecule glycomimetics against free fatty acid­induced endothelial dysfunction, focusing on nitric oxide (NO) and oxidative stress pathways. Methods: Four glycomimetics were synthesized by the stepwise transformation of 2,5­dihydroxybenzoic acid to a range of 2,5­substituted benzoic acid derivatives, incorporating the key sulfate groups to mimic the interactions of heparan sulfate. Endothelial function was assessed using acetylcholine­induced, endotheliumdependent relaxation in mouse thoracic aortic rings using wire myography. Human umbilical vein endothelial cell (HUVEC) behavior was evaluated in the presence or absence of the free fatty acid, palmitate, with or without glycomimetics (1µM). DAF­2 and H2DCF­DA assays were used to determine nitric oxide (NO) and reactive oxygen species (ROS) production, respectively. Lipid peroxidation colorimetric and antioxidant enzyme activity assays were also carried out. RT­PCR and western blotting were utilized to measure Akt, eNOS, Nrf­2, NQO­1 and HO­1 expression. Results: Ex vivo endothelium­dependent relaxation was significantly improved by the glycomimetics under palmitate­induced oxidative stress. In vitro studies showed that the glycomimetics protected HUVECs against the palmitate­induced oxidative stress and enhanced NO production. We demonstrate that the protective effects of pre­incubation with glycomimetics occurred via upregulation of Akt/eNOS signaling, activation of the Nrf2/ARE pathway, and suppression of ROS­induced lipid peroxidation. Conclusion: We have developed a novel set of small molecule glycomimetics that protect against free fatty acidinduced endothelial dysfunction and thus, represent a new category of therapeutic drugs to target endothelial damage, the first line of defense against cardiovascular disease

DNA conformational effects on the interaction of netropsin with A-tract sequences

The influence of cosolutes and DNA sequence on the interaction of netropsin with three duplexes has been studied by isothermal titration calorimetry. In buffer, netropsin forms two complexes with a net stoichiometry of 1:1 in the minor groove of the oligonucleotide (GCGCGAATTCGCGC)2. One complex has a weaker affinity and is more enthalpically favored relative to the other one, consistent with previous studies Freyer, M. W., et al. (2006) Biophys. Chem. 126, 186-196]. With the cosolutes betaine and 2-methyl-2,4-pentanediol, the enthalpy and heat capacity changes indicate that the complex with weaker affinity is disfavored relative to the complex with higher affinity. With (CGCGCAATTGCGCG)2, netropsin has one binding mode in buffer, and complex formation is not influenced by the cosolutes. The similarities of the enthalpy and heat capacity changes suggest that netropsin interacts similarly with these two oligonucleotides in the presence of cosolutes. The oligonucleotide (GCGCAAATTTGCGC)2 also forms two complexes with netropsin, and the complex with weaker affinity is again disfavored by the cosolutes. Thus, the interaction of netropsin with these A/T binding sites is influenced both by the bases adjacent to the binding site and by cosolutes. We suggest that these two factors influence the conformation of the minor-groove binding site of DNA

Supercooling along an altitudinal gradient in Espeletia schulzii a caulescent giant rosette species

Publicado en: Journal of Experimental Botany Vol. 38: pp [email protected]@ula.v

Estudio morfoanatómico de dos orquídeas de una selva nublada tropical

Se describen por primera vez la estructura morfo-anatómica y estrategias hídricas de dos orquídeas de las selvas nubladas andinas de Venezuela, Maxillaria miniata y Pleurothallis cardiantha. El estudio se efectuó en una selva nublada poco intervenida y en el bosque secundario del Jardín Botánico de la Universidad de los Andes, Venezuela. Las muestras recolectadas fueron fijadas en FAA y sometidas a métodos tradicionales de anatomía vegetal y de MEB. Se determinaron sus potenciales hídricos foliares durante el año 2004-2005, para determinar si presentan un comportamiento CAM típico, CAM facultativo o C3. Ambas especies exhiben caracteres xeromórficos en sus órganos vegetativos. M. miniata presenta seudobulbos como principales reservorios hídricos, hojas delgadas cartáceas, células epidérmicas de paredes engrosadas, hipodermis adaxial biestratificada y abaxial uniestratificada, clorénquima homogéneo con haces bien desarrollados, abundante esclerénquima, y cristales silíceos asociados a tejidos epidérmicos y esclerenquimáticos. En contraste, P. cardiantha presenta hojas suculentas a coriáceas, epidermis adaxial papilosa, mesófilo heterogéneo, haces pequeños, poco desarrollados e hipodermis acuífera pluriestratificada en ambas caras, que junto con los idioblastos acuíferos del clorénquima constituyen los tejidos de reserva de agua. Los potenciales hídricos registrados en estas especies se mantuvieron elevados y casi invariables durante todo el año, lo cual sugiere un metabolismo CAM en ambos [email protected], [email protected]@[email protected] morpho-anatomical vegetative structure and water-storing strategies of two orchids of the Venezuelan Cloud Forests,Pleurothallis cardiantha and Maxillaria miniata, are described for the first time. Field work was conducted in a relatively preserved Cloud Forest and a secondary forest of the Botanical Garden of the Universidad de los Andes, Venezuela, located in the same vicinity. Botanical specimens were stored in FAA and submitted to standard plant anatomy microtechnique procedures, including SEM. Monthly leaf-water potentials were recorded during 2004-2005 in order to determine whether these species behave as typical CAM, facultative CAM or C3 species. Both exhibit xeromorphic characters in their vegetative organs. M. mineata presents pseudobulbs as main water storing structures, thin cartaceous leaves, epidermal and sclerenchymatic tissues associated with siliceous crystals, adaxial hypodermis double layered, single-layered abaxial hypodermis, homogenous mesophyll with well developed collateral bundles, and abundant sclerenchyma. P. cardiantha, in contrast, presents moderately succulent to coriaceous leaves, adaxial epidermal cell forming papillae, heterogenous mesophyll with poorly developed vascular bundles. The water-storing tissues in this species involve the adaxial and abaxial hypodermis, as well as the acuiferous idioblasts located in the spongy parenchyma. The leaf waterpotentials registered were high and remained practically invariable during the year, which suggests a typical CAM metabolism in both cases

Resistencia a glifosato en raigrás anual en Uruguay. ¿La importamos o la generamos en el país?

En este artículo se presentan los avances del proyecto ?Resistencia a glifosato en raigrás anual en Uruguay ? entender, reducir y prevenir?, que tiene como objetivos principales determinar si las poblaciones resistentes fueron importadas, se seleccionaron localmente, o son el resultado de ambos procesos, y cuál es el grado y riesgo de dispersión de la resistencia a través del uso de las variedades utilizadas en la producción de forraje