Ruolo dell'Enantiomero (R)-9-HSA nel controllo della proliferazione in una linea di Adenocarcinoma del Colon umano

9-hydroxystearic acid (9-HSA) is an endogenous lipoperoxidation product and its administration to HT29, a colon adenocarcinoma cell line, induced a proliferative arrest in G0/G1 phase mediated by a direct activation of the p21WAF1 gene, bypassing p53. We have previously shown that 9-HSA controls cell growth and differentiation by inhibiting histone deacetylase 1 (HDAC1) activity, showing interesting features as a new anticancer drug. The interaction of 9-HSA with the catalytic site of the 3D model has been tested with a docking procedure: noticeably, when interacting with the site, the (R)-9-enantiomer is more stable than the (S) one. Thus, in this study, (R)- and (S)-9-HSA were synthesized and their biological activity tested in HT29 cells. At the concentration of 50 M (R)-9-HSA showed a stronger antiproliferative effect than the (S) isomer, as indicated by the growth arrest in G0/G1. The inhibitory effect of (S)-9-HSA on HDAC1, HDAC2 and HDAC3 activity was less effective than that of the (R)-9-HSA in vitro, and the inhibitory activity of both the (R)- and the (S)-9-HSA isomer, was higher on HDAC1 compared to HDAC2 and HDAC3, thus demonstrating the stereospecific and selective interaction of 9-HSA with HDAC1. In addition, histone hyperacetylation caused by 9-HSA treatment was examined by an innovative HPLC/ESI/MS method. Analysis on histones isolated from control and treated HT29 confirmed the higher potency of (R)-9-HSA compared to (S)-9-HSA, severely affecting H2A-2 and H4 acetylation. On the other side, it seemed of interest to determine whether the G0/G1 arrest of HT29 cell proliferation could be bypassed by the stimulation with the growth factor EGF. Our results showed that 9-HSA-treated cells were not only prevented from proliferating, but also showed a decreased [3H]thymidine incorporation after EGF stimulation. In this condition, HT29 cells expressed very low levels of cyclin D1, that didn’t colocalize with HDAC1. These results suggested that the cyclin D1/HDAC1 complex is required for proliferation. Furthermore, in the effort of understanding the possible mechanisms of this effect, we have analyzed the degree of internalization of the EGF/EGFR complex and its interactions with HDAC1. EGF/EGFR/HDAC1 complex quantitatively increases in 9-HSA-treated cells but not in serum starved cells after EGF stimulation. Our data suggested that 9-HSA interaction with the catalytic site of the HDAC1 disrupts the HDAC1/cyclin D1 complex and favors EGF/EGFR recruitment by HDAC1, thus enhancing 9-HSA antiproliferative effects. In conclusion 9-HSA is a promising HDAC inhibitor with high selectivity and specificity, capable of inducing cell cycle arrest and histone hyperacetylation, but also able to modulate HDAC1 protein interaction. All these aspects may contribute to the potency of this new antitumor agent.

Role of (R)-9-hydroxystearic acid in zebrafish development

9-hydroxystearic acid (9-HSA) belongs to a class of lipid peroxidation products identified in several human and murine cell lines. These products are greatly diminished in tumors compared to normal tissues and their amount is inversely correlated with the malignancy of the tumor. 9-HSA activity has been tested in cancer cell lines, where it showed to act as a histone deacetylase 1 (HDAC1) inhibitor. In particular, in a colon cancer cell line (HT29), its administration resulted in an inhibition of proliferation together with an induction of differentiation. In this thesis the effect of (R)-9-hydroxystearic acid has been tested in vivo on cell proliferation and differentiation processes, in the early stages of zebrafish development. The final aim of this work was to elucidate the role of (R)-9-HSA in the control of cell differentiation and proliferation during normal development, in order to better understand its molecular control of cancerogenesis. The molecule has been administered via injection in the yolk of zebrafish embryos. The analysis of the histone acetylation pattern showed a hyperacetilation of histone H4 after treatment with the molecule, as detectable in HDAC1 mutants. (R)-9-HSA was also demonstrated to interfere with the signaling pathways that regulate proliferation and differentiation in zebrafish retina and hindbrain. This resulted in a reduction of proliferation in the hindbrain at 24 hours post injection (hpi), and in a hyperproliferation at 48 and 72 hpi in the retina, with a concomitant inhibition of differentiation. Finally, (R)-9-HSA effects were evident on proliferation of stem cell located in the ciliary marginal zone (CMZ) of the retina. The presence of ROS and 4-hydroxynoneal in the CMZ of wild-type embryos supports the hypothesis that oxidative stress could regulate stem cells fate in zebrafish retina.

Propuesta de metodología para la calificación bio-ambiental de espacios verdes mediante coeficientes ecofisiológicos

Uno de los mayores problemas que enfrentan las urbes modernas es su crecimiento horizontal en base a cemento y asfalto, cubriendo grandes extensiones. Su resultado es un importante incremento de la temperatura llamado efecto "isla de calor", aspecto de particular importancia en años venideros por el creciente calentamiento global, a lo cual se añade la contaminación por gases y partículas. Las áreas destinadas a mitigar parcialmente estos efectos son los denominados espacios verdes, los cuales representan una mancha en el mapa. Sin embargo, su impacto termodinámico a nivel ambiental desde el punto de vista del confort y calidad de vida humanos puede ser muy variable según como esté diseñado. Este trabajo pretende establecer una metodología de aplicación en cualquier región, para calificar cuantitativamente el valor de los diferentes tipos de espacios verdes como modificadores ambientales, ya sean ejecutados o a nivel de proyecto. A través de esta metodología se determinan índices o coeficientes regionales objetivos y de sencilla aplicación. Para desarrollar estos índices, a los espacios verdes se aplican, adaptados, principios de la termodinámica de superficies húmedas y de la ecofisiología. A través de procedimientos matemáticos y conceptos fisiológicos se concluye en fórmulas para calcular índices regionales de aplicación directa en la evaluación de proyectos. A modo de comprobación de la metodología se determina el valor ambiental comparativo de diferentes tipos de espacios verdes correspondientes al sistema integrado de espacios verdes de la ciudad de Mendoza

Pigment data set from the SeaWiFS Bio-optical Archive and Storage System (1961-1996)

The oceans play a critical role in the Earth's climate, but unfortunately, the extent of this role is only partially understood. One major obstacle is the difficulty associated with making high-quality, globally distributed observations, a feat that is nearly impossible using only ships and other ocean-based platforms. The data collected by satellite-borne ocean color instruments, however, provide environmental scientists a synoptic look at the productivity and variability of the Earth's oceans and atmosphere, respectively, on high-resolution temporal and spatial scales. Three such instruments, the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) onboard ORBIMAGE's OrbView-2 satellite, and two Moderate Resolution Imaging Spectroradiometers (MODIS) onboard the National Aeronautic and Space Administration's (NASA) Terra and Aqua satellites, have been in continuous operation since September 1997, February 2000, and June 2002, respectively. To facilitate the assembly of a suitably accurate data set for climate research, members of the NASA Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project and SeaWiFS Project Offices devote significant attention to the calibration and validation of these and other ocean color instruments. This article briefly presents results from the SIMBIOS and SeaWiFS Project Office's (SSPO) satellite ocean color validation activities and describes the SeaWiFS Bio-optical Archive and Storage System (SeaBASS), a state-of-the-art system for archiving, cataloging, and distributing the in situ data used in these activities.

Bio- and magnetostratigraphic events of ODP holes of the central Kerguelen Plateau

Neogene biostratigraphic and magnetostratigraphic data are compiled from Holes 747A, 748B, and 751A drilled on the Southern Kerguelen Plateau during Ocean Drilling Program Leg 120. Neogene sections have excellent to good magnetostratigraphic signatures in many intervals. This, in addition to minimal coring gaps and the occurrence of mixed assemblages of both calcareous and siliceous microfossil assemblages, makes these valuable biostratigraphic reference sections for intra- and extraregional correlations. This paper combines the sequence of biostratigraphic events reported from diatom, radiolarian, planktonic foraminifer, calcareous nannofossil, and silicoflagellate studies of Leg 120 sediments. It correlates microfossil datums with the geomagnetic polarity time scale to test existing age estimates and to refine biostratigraphic age controls for the southern high latitudes. Significant biostratigraphic datums are presented in a series of age-depth plots. Numerous hiatuses are clearly identified through this approach, and the positions of lesser disconformities are suggested. Most Neogene intervals are represented in at least one site, although "regional" unconformities occur in the upper Pliocene, uppermost Miocene/lowermost Pliocene, middle upper Miocene, middle middle Miocene, and at the lower/middle Miocene boundaries. The longest hiatus spanned 6 m.y., with most other hiatuses representing 1 m.y. or less. This paper compiles Leg 120 biostratigraphic and magnetostratigraphic data for use in future syntheses of southern high latitude biostratigraphy and presents an age model for Leg 120 Neogene sediments.