81 resultados para ROS e dano ao DNA


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Se han calibrado, validado y comparado tres modelos de resistencia al flujo de contorno granular: un modelo potencial y otros dos modelos desarrollados para condiciones de alta rugosidad relativa (uno basado en una modificación de la ley logarítmica de Prandtl-von Karman y otro fundamentado en un perfil de velocidad configurado en dos zonas: una uniforme en las proximidades de los elementos de rugosidad y otra superior que sigue una distribución logarítmica). Se ha empleado para ello un numeroso conjunto de 1.533 datos tomados en ríos y en canales de laboratorio, representativo de un amplio intervalo hidráulico y geomorfológico en el ámbito de ríos de grava y de montaña. Han resultado preferibles las ecuaciones ajustadas con los percentiles granulométricos mayores (d90 o d84) que las ajustadas con el diámetro mediano (d50), debido a la mayor capacidad explicativa alcanzada dado un modelo, la menor diferencia en la bondad de ajuste entre los diferentes modelos y la menor influencia del origen de los datos (río o canal de laboratorio). Las ecuaciones ajustadas de acuerdo con los modelos en donde se contemplan condiciones de alta rugosidad relativa presentan predicciones similares, exceptuando el intervalo macrorrugoso (y/d90 < 1), en el que es preferible la correspondiente al modelo fundamentado en el perfil de velocidad configurado en dos zonas. Se recomienda restringir la aplicación de la ecuación ajustada con arreglo a la ley potencial al intervalo de y/d90 comprendido entre uno y veinte, puesto que fuera de dicho intervalo tiende a infraestimar notablemente la resistencia al flujo.

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Con base en una selección de 145 datos pertenecientes a ríos de montaña de fuerte pendiente (entre el 1 y 5%), de alineación no sinuosa, sin vegetación significativa en el cauce y sin transporte intenso de sedimentos, se han desarrollado un conjunto de eucaciones en régimen permanente y uniforme que no requieren la estimación independiente de coeficiente de resistencia alguno, con la subsiguiente reducción en coste y esfuerzo de cálculo. Siempre que se observe estrictamente el rango experimental en el que rigen, las ecuaciones derivadas permiten una predicción de precisión equiparable a los métodos basados en la determinacion de un coeficiente de resistencia adecuado para el tramo de estudio.

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Con base en un conjunto de 904 datos hidrométricos pertenecientes a ríos de grava (de alienación no sinuosa desprovistos de cubierta vegetal significativa en el cauce y carentes de obstáculos) se han desarrollado un conjunto de ecuaciones en régimen uniforme que no requieren la estimación independiente de un coeficiente de resistencia, con la subsiguiente reducción de coste y esfuerzo de cálculo. Siempre que se respete estrictamente el rango experimental en el que rigen, las ecuaciones derivadas permiten una predicción de precisión equiparable a la de aquellos métodos basados en la determinación de un coeficiente de resistencia adecuado para el tramo de estudio.

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Con base en una selección de 145 datos pertenecientes a ríos de montaña de fuerte pendiente (³ 1%) se han desarrollado cinco expresiones para determinar el factor de fricción de Darcy-Weisbach. La primera expresión se fundamenta en la aplicación para flujo turbulento rugoso en lámina libre de la ley semilogarítmica de Prandtl-Kárman, que es función de la sumersión relativa (relación entre el calado medio y la rugosidad equivalente). La segunda y tercera corresponden a correciones de la primera para flujo macrorrugoso, propuestas por Thompson y Campbell (1979) y Aguirre-Pe y Fuentes (1990) respectivamente. La cuarta ecuación consiste una en potencia de la sumersión relativa, mientras que la quinta corrige la fórmula anterior incorporando una potencia de la pendiente, tal y como propugnan Meunier (1989) y Rickenmann (1990). Las expresiones derivadas presentan un ajuste significativo, si se tienen en cuenta las limitaciones hidrométricas existentes en ríos de material grueso y fuerte pendiente. Destaca el mayor ajuste conseguido con las ecuaciones con las ecuaciones del tipo potencial frente a las del tipo semilogarítmico. Se ha encontrado, asimismo, una capacidad de predicción ligeramente superior en aquellas expresiones que incluyen modificaciones respecto a la ecuación original, ecuaciones del tipo segundo, tercero y quinto anteriormente indicado.

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El concepto de aspereza equivalente (ks) permite relacionar la resistencia al flujo de cauces de contorno granular con el tamaño de las partículas que lo conforman, mediante la ley logarítmica de distribución vertical de velocidad de la corriente. En este artículo se revisa el estado del conocimiento acerca de los criterios para la predicción de dicha relación con el objetivo de brindar una guía para la selección de los mismos. Pese a que el análisis de las experiencias compiladas desvela la dispersión de resultados, a efectos prácticos se recomienda para ríos de grava y de montaña: ks 3•d90; ks 3,4•d84 y ks 7•d50. Dicha dispersión y el hecho de que ks sea varias veces superior al diámetro medio del sedimento puede atribuirse en diferente grado: a la heterogeneidad del sedimento en lechos naturales, a las formas de fondo, al transporte sólido de fondo,así como a las limitaciones del modelo logarítmico de distribución de velocidad bajo ciertas condiciones de flujo. La predicción de ks en función de variables estadísticas del campo de elevaciones del lecho se ha revelado como una alternativa con una elevada capacidad explicativa, por lo que en la medida que avancen las técnicas microtopográficas se consolidará esta vía como el método del futuro.

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DnaSP, DNA Sequence Polymorphism, is a software package for the analysis of nucleotide polymorphism from aligned DNA sequence data. DnaSP can estimate several measures of DNA sequence variation within and between populations (in noncoding, synonymous or nonsynonymous sites, or in various sorts of codon positions), as well as linkage disequilibrium, recombination, gene flow and gene conversion parameters. DnaSP can also carry out several tests of neutrality: Hudson, Kreitman and Aguadé (1987), Tajima (1989), McDonald and Kreitman (1991), Fu and Li (1993), and Fu (1997) tests. Additionally, DnaSP can estimate the confidence intervals of some test-statistics by the coalescent. The results of the analyses are displayed on tabular and graphic form.

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Four new metal complexes {M = Pd(II) or Pt(II)} containing the ligand 9-aminoacridine (9AA) were prepared. The compounds were characterized by FT-IR and 1H, 13C, and 195Pt NMR spectroscopies. Crystal structure of the palladium complex of formulae [Pd(9AA)(μ-Cl)]2 · 2DMF was determined by X-ray diffraction. Two 9-acridine molecules in the imine form bind symmetrically to the metal ions in a bidentate fashion through the imine nitrogen atom and the C(1) atom of the aminoacridine closing a new five-membered ring. By reaction with phosphine or pyridine, the Cl bridges broke and compounds with general formulae [Pd(9AA)Cl(L)] (where L = PPh3 or py) were formed. A mononuclear complex of platinum of formulae [Pt(9AA)Cl(DMSO)] was also obtained by direct reaction of 9-aminoacridine and the complex [PtCl2(DMSO)2]. The capacity of the compounds to modify the secondary and tertiary structures of DNA was evaluated by means of circular dichroism and electrophoretic mobility. Both palladium and platinum compounds proved active in the modification of both the secondary and tertiary DNA structures. AFM images showed noticeable modifications of the morphology of the plasmid pBR322 DNA by the compounds probably due to the intercalation of the complexes between base pairs of the DNA molecule. Finally, the palladium complex was tested for antiproliferative activity against three different human tumor cell lines. The results suggest that the palladium complex of formula [Pd(9AA)(μ-Cl)]2 has significant antiproliferative activity, although it is less active than cisplatin.

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Four new metal complexes {M = Pd(II) or Pt(II)} containing the ligand 9-aminoacridine (9AA) were prepared. The compounds were characterized by FT-IR and 1H, 13C, and 195Pt NMR spectroscopies. Crystal structure of the palladium complex of formulae [Pd(9AA)(μ-Cl)]2 · 2DMF was determined by X-ray diffraction. Two 9-acridine molecules in the imine form bind symmetrically to the metal ions in a bidentate fashion through the imine nitrogen atom and the C(1) atom of the aminoacridine closing a new five-membered ring. By reaction with phosphine or pyridine, the Cl bridges broke and compounds with general formulae [Pd(9AA)Cl(L)] (where L = PPh3 or py) were formed. A mononuclear complex of platinum of formulae [Pt(9AA)Cl(DMSO)] was also obtained by direct reaction of 9-aminoacridine and the complex [PtCl2(DMSO)2]. The capacity of the compounds to modify the secondary and tertiary structures of DNA was evaluated by means of circular dichroism and electrophoretic mobility. Both palladium and platinum compounds proved active in the modification of both the secondary and tertiary DNA structures. AFM images showed noticeable modifications of the morphology of the plasmid pBR322 DNA by the compounds probably due to the intercalation of the complexes between base pairs of the DNA molecule. Finally, the palladium complex was tested for antiproliferative activity against three different human tumor cell lines. The results suggest that the palladium complex of formula [Pd(9AA)(μ-Cl)]2 has significant antiproliferative activity, although it is less active than cisplatin.

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Four new metal complexes {M = Pd(II) or Pt(II)} containing the ligand 9-aminoacridine (9AA) were prepared. The compounds were characterized by FT-IR and 1H, 13C, and 195Pt NMR spectroscopies. Crystal structure of the palladium complex of formulae [Pd(9AA)(μ-Cl)]2 · 2DMF was determined by X-ray diffraction. Two 9-acridine molecules in the imine form bind symmetrically to the metal ions in a bidentate fashion through the imine nitrogen atom and the C(1) atom of the aminoacridine closing a new five-membered ring. By reaction with phosphine or pyridine, the Cl bridges broke and compounds with general formulae [Pd(9AA)Cl(L)] (where L = PPh3 or py) were formed. A mononuclear complex of platinum of formulae [Pt(9AA)Cl(DMSO)] was also obtained by direct reaction of 9-aminoacridine and the complex [PtCl2(DMSO)2]. The capacity of the compounds to modify the secondary and tertiary structures of DNA was evaluated by means of circular dichroism and electrophoretic mobility. Both palladium and platinum compounds proved active in the modification of both the secondary and tertiary DNA structures. AFM images showed noticeable modifications of the morphology of the plasmid pBR322 DNA by the compounds probably due to the intercalation of the complexes between base pairs of the DNA molecule. Finally, the palladium complex was tested for antiproliferative activity against three different human tumor cell lines. The results suggest that the palladium complex of formula [Pd(9AA)(μ-Cl)]2 has significant antiproliferative activity, although it is less active than cisplatin.

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A photoactivated ruthenium(II) arene complex has been conjugated to two receptor-binding peptides, a dicarba analogue of octreotide and the Arg-Gly-Asp (RGD) tripeptide. These peptides can act as"tumor-targeting devices" since their receptors are overexpressed on the membranes of tumor cells. Both ruthenium-peptide conjugates are stable in aqueous solution in the dark, but upon irradiation with visible light, the pyridyl-derivatized peptides were selectively photodissociated from the ruthenium complex, as inferred by UV-vis and NMR spectroscopy. Importantly, the reactive aqua species generated from the conjugates, [(η6-p-cym)Ru(bpm)(H2O)]2+, reacted with the model DNA nucleobase 9-ethylguanine as well as with guanines of two DNA sequences, 5′dCATGGCT and 5′dAGCCATG. Interestingly, when irradiation was performed in the presence of the oligonucleotides, a new ruthenium adduct involving both guanines was formed as a consequence of the photodriven loss of p-cymene from the two monofunctional adducts. The release of the arene ligand and the formation of a ruthenated product with a multidentate binding mode might have important implications for the biological activity of such photoactivated ruthenium(II) arene complexes. Finally, photoreactions with the peptide-oligonucleotide hybrid, Phac-His-Gly-Met-linker-p5′dCATGGCT, also led to arene release and to guanine adducts, including a GG chelate. The lack of interaction with the peptide fragment confirms the preference of such organometallic ruthenium(II) complexes for guanine over other potential biological ligands, such as histidine or methionine amino acids.

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Nuclear DNA content in gametophytes and sporophytes or the prostrate phases of the following species of Bonnemaisoniaceae (Asparagopsis armata, Asparagopsis taxiformis, Bonnemaisonia asparagoides, Bonnemaisonia clavata and Bonnemaisonia hamifera) were estimated by image analysis and static microspectrophotometry using the DNA-localizing fluorochrome DAPI (4′, 6-diamidino-2-phenylindole, dilactate) and the chicken erythrocytes standard. These estimates expand on the Kew database of DNA nuclear content. DNA content values for 1C nuclei in the gametophytes (spermatia and vegetative cells) range from 0.5 pg to 0.8 pg, and for 2C nuclei in the sporophytes or the prostrate phases range from 1.15-1.7 pg. Although only the 2C and 4C values were observed in the sporophyte or the prostrate phase, in the vegetative cells of the gametophyte the values oscillated from 1C to 4C, showing the possible start of endopolyploidy. The results confirm the alternation of nuclear phases in these Bonnemaisoniaceae species, in those that have tetrasporogenesis, as well as those that have somatic meiosis. The availability of a consensus phylogenetic tree for Bonnemaisoniaceae has opened the way to determine evolutionary trends in DNA contents. Both the estimated genome sizes and the published chromosome numbers for Bonnemaisoniaceae suggest a narrow range of values consistent with the conservation of an ancestral genome.

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Source/Description: p6a-l is a O.9 kb HindIII/BamHl genomic fragment subclone or cosmic cNX.6a in pUC13. cNX.6a was isolated from a non-methylated enriched library from the CMGT cell line Cll (1,2).

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ABSTRACT Proposal for an evaluation protocol of the ecological quality of Andean rivers (CERA) and its use in two basins in Ecuador and Peru A Rapid Protocol is presented for Evaluation of the Ecological Status of Andean Rivers (CERA) localized over 2000 m.a.s.l. from the Northern Andes (Venezuela) through the Altiplano in the Central Andes (Bolivia). This protocol was used in 45 sampling sites in the Guayllabamba River Basin in Ecuador and in 42 sampling sites in the Ca nete River Basin in Peru. Previously, and in order to test if the sampling stations may or not be considered reference stations, we constructed a method that assesses 24 basin attributes, hydrology, reach and riverbed and that uctuates from 24 to 120 points; sites with values higher than 100 were considered as potential reference sites. Besides the benthic macroinvertebrats" evaluation, the river habitat and riparian vegetation were also evaluated through of the application of the indices ABI (R´ os et al., submitted), IHF (Pardo et al., 2002) and QBR-And, respectively. The convenience of the initial allocation of the reference sites was evaluated as well. These indices have been properly adapted to the conditions and characteristics of the high Andes rivers. The results obtained for both basins were compared and discussed. Through the use of the CERA protocol, the particular perturbation gradients and the natural variability of the reference sites in both countries were recognized. RESUMEN Propuesta de un protocolo de evaluación de la calidad ecológica de ríos andinos (CERA) y su aplicaci´on a dos cuencas en Ecuador y Perú Se presenta un protocolo rápido de evaluación de la Calidad Ecológica de Ríos Andinos (CERA), situados sobre los 2000 m.s.n.m, desde los Andes del Norte (Venezuela) hasta el Altiplano de los Andes Centrales (Bolivia). Este protocolo ha sido aplicado en 45 estaciones de muestreo en la cuenca del río Guayllabamba en Ecuador y en 42 estaciones de muestreo en la cuenca del río Cañete en Perú. Previamente, para probar si las estaciones de muestreo pueden o no ser estaciones de referencia construimos un método que valora 24 atributos de cuenca, hidrología, tramo y lecho y que fluctúa de 24 a 120 puntos; valores superiores a 100 fueron considerados como sitios potencialmente de referencia. Además del estudio de los macroinvertebrados bentónicos, se evaluó el hábitat fluvial y la comunidad vegetal de ribera a través de la aplicación de los índices ABI (Ríos et al., sometido), IHF (Pardo et al., 2002) y QBR-And respectivamente; así como la conveniencia de la asignación inicial de las estaciones de referencia. Estos índices han sido adecuadamente adaptados a las condiciones y características propias de los ríos altoandinos. Los resultados obtenidos fueron comparados y discutidos entre ambas cuencas. Mediante la aplicación del protocolo CERA se han reconocido los respectivos gradientes de perturbación y la variabilidad natural de las estaciones de referencia en ambos países.

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The nucleoid-associated protein H-NS is a global modulator of the expression of genes associated with adaptation to environmental changes. A variant of H-NS expressed in the R27 plasmid was previously shown to selectively modulate the expression of horizontally acquired genes, with minimal effects on core genes that are repressed by the chromosomal form of H-NS. Both H-NS proteins are formed by an oligomerization domain and a DNA-binding domain, which are connected by a linker that is highly flexible in the absence of DNA. We studied DNA binding by means of oligomer-forming chimeric proteins in which domains of the chromosomal and plasmidic variants are exchanged, as well as in monomeric truncated forms containing the DNA-binding domain and variable portions of the linker. Point mutations in the linker were also examined in full-length and truncated H-NS constructs. These experiments show that the linker region contributes to DNA binding affinity and that it is a main component of the distinct DNA binding properties of chromosomal and plasmidic H-NS. We propose that interactions between the linker and DNA limit the flexibility of the connection between H- NS oligomerization and DNA binding and provide an allosteric indirect readout mechanism to detect long- range distortions of DNA, thus enabling discrimination between core and horizontally acquired DNA.

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Mitochondrial DNA (mtDNA), a maternally inherited 16.6-Kb molecule crucial for energy production, is implicated in numerous human traits and disorders. It has been hypothesized that the presence of mutations in the mtDNA may contribute to the complex genetic basis of schizophreniadisease, due to the evidence of maternal inheritance and the presence of schizophrenia symptoms in patients affected of a mitochondrial disorder related to a mtDNA mutation. The present project aims to study the association of variants of mitochondrial DNA (mtDNA), and an increased risk of schizophrenia in a cohort of patients and controls from the same population. The entire mtDNA of 55 schizophrenia patients with an apparent maternal transmission of the disease and 38 controls was sequenced by Next Generation Sequencing (Ion Torrent PGM, Life Technologies) and compared to the reference sequence. The current method for establishing mtDNA haplotypes is Sanger sequencing, which is laborious, timeconsuming, and expensive. With the emergence of Next Generation Sequencing technologies, this sequencing process can be much more quickly and cost-efficiently. We have identified 14 variants that have not been previously reported. Two of them were missense variants: MTATP6 p.V113M and MTND5 p.F334L ,and also three variants encoding rRNA and one variant encoding tRNA. Not significant differences have been found in the number of variants between the two groups. We found that the sequence alignment algorithm employed to align NGS reads played a significant role in the analysis of the data and the resulting mtDNA haplotypes. Further development of the bioinformatics analysis and annotation step would be desirable to facilitate the application of NGS in mtDNA analysis.