Avoiding acidic region streaking in two-dimensional gel electrophoresis: Case study with two bacterial whole cell protein extracts

Acidic region streaking (ARS) is one of the lacunae in two-dimensional gel electrophoresis (2DE) of bacterial proteome. This streaking is primarily caused by nucleic acid (NuA) contamination and poses major problem in the downstream processes like image analysis and protein identification. Although cleanup and nuclease digestion are practiced as remedial options, these strategies may incur loss in protein recovery and perform incomplete removal of NuA. As a result, ARS has remained a common observation across publications, including the recent ones. In this work, we demonstrate how ultrasound wave can be used to shear NuA in plain ice-cooled water, facilitating the elimination of ARS in the 2DE gels without the need for any additional sample cleanup tasks. In combination with a suitable buffer recipe, IEF program and frequent paper-wick changing approach, we are able to reproducibly demonstrate the production of clean 2DE gels with improved protein recovery and negligible or no ARS. We illustrate our procedure using whole cell protein extracts from two diverse organisms, Escherichia coli and Mycobacterium smegmatis. Our designed protocols are straightforward and expected to provide good 2DE gels without ARS, with comparable times and significantly lower cost.

Prudentia Iuris, 1981, n° 04 (número completo)

Contenido: Editorial – Jorge Joaquín Llambías : 18-VIII-1981 / Jorge A. Mazzinghi – El orden civil / Tomás D. Casares – La prudencia jurídica : sobre el conocimiento de lo justo concreto / Carlos Ignacio Massini –El derecho a la vida y sus impicancias respecto de la pena de muerte y el aborto a la luz del derecho natural / Silvio Pablo Pestalardo – Pluralismo y sociedad / Fulvio Ramos – Una definición romana del derecho : ars boni et aequi / Edmundo J. Carbone – Hacia la reforma de la ley de adopción / Jorge A. Mazzinghi – La prodigalidad en la Suma Teológica y en la reforma del Código Civil Argentino / Javier Reigada – Escolios – Documentos – Notas bibliográficas

Retórica y música barroca : una posible hermenéutica

Resumen: Este trabajo considera algunos conceptos retóricos expuestos por los teóricos de la musica poetica y por otras fuentes, útiles para la formación de intérpretes dedicados a los repertorios de los siglos XVII y XVIII.

La poética del relato de viaje entre la Edad Media y el siglo XXI

Resumen: Este ensayo reproduce la conferencia plenaria pronunciada en las Undécimas Jornadas Internacionales de Literatura Española Medieval en la Pontificia Universidad Católica Argentina. Pone en relación la narrativa de viajes de la Edad Media con su versión contemporánea en el mundo hispánico: después de trazar un panorama general, presenta una selección de rasgos que se mantienen sin gran variación hasta los siglos XX y XXI (doble temporalidad, presencia del azar, recursos retóricos, variaciones de lo maravilloso, etc.). En la segunda parte trata la relación con las disciplinas científicas, el relato de viaje como género, el desplazamiento, la ilustración fotográfica y la complejidad de la edición, entre otros puntos. El ensayo concluye con unas breves reflexiones sobre la valoración del relato de viaje como género literario.

El viaje dantesco y su funcionalidad metapoética en El sueño y el Infierno de los enamorados de Santillana

Resumen: El sueño y el Infierno de los enamorados del Marqués de Santillana están construidos, como sabemos, siguiendo el modelo del viaje alegórico dantesco. Llama la atención, sin embargo, la alta concentración de recursos metapoéticos que presentan estos decires narrativos en comparación a lo esporádicos que resultan en la Commedia, lo cual nos ha permitido inferir que los mismos tendrán un papel fundamental en cuanto clave interpretativa del propio texto. Por tanto, intentaremos desarrollar aquí de qué manera se van articulando y conectando las acciones del ver, el leer y el escribir (en sus variantes: relatar, recontar, metrificar, glosar, etc.), de modo de formar un verdadero sistema metapoético que permitiría interpretar al viaje onírico santillanesco como un viaje de lectura, de escritura y de reflexión literaria.

Memoria poética y geografía en Don Juan Manuel

Integran este número de la revista ponencias presentadas en Studia Hispanica Medievalia VIII: Actas de las IX Jornadas Internacionales de Literatura Española Medieval, 2008, y de Homenaje al Quinto Centenario de Amadis de Gaula.

La aristocracia de Castilla y sus abogados celestiales

Resumen: Este trabajo se ocupa de la relación entre la aristocracia de Castilla y los santos, como protectores en vida y, sobre todo, en el momento de la muerte. A partir de las cláusulas testamentarios y de las recomendaciones recogidas en los tratados del género Ars moriendi, señala las diferencias entre los vínculos que mantienen con el mundo celestial las élites de poder y el resto de la sociedad. Analiza, igualmente, los criterios para la elección de determinados santos como intercesores. Estos se basan en la primacía otorgada al entorno de Cristo, en la petición de auxilio a los bienaventurados que ayudan a los que van a dejar este mundo a conseguir la salvación del alma, y también a los defensores del grupo caballeresco y los titulares de iglesias y monasterios vinculados al señor.

Un testigo directo de las tensiones religiosas en la Alejandría de los siglos IV y V D.C. : Páladas de Alejandría en la Antología Palatina

Abstract: Although he is the most prolific writer of the Greek Anthology, Palladas’ life is almost unknown. But, in spite of the limited biographical data, his work has fortunately been preserved in the GA. Because of his literary creation, the old times rewarded him with the nickname Μετέωρος, high, since the literary merits of a hundred poems of his authorship was recognized (others are discussed by critics). A dozen of them contains invaluable information about the political, religious and social situation in Alexandria immediately after the victory of the bishop Theophilus, Cyril’s predecessor and uncle, during the conflict between Christians and Gentiles, each sector backed up by an Alexandrian population segment (cfr. 9.528, 10.82, 10.85) . Resigned at times, constantly demanding, Palladas complains about the decline of the belief professed by him due to the increasing penetration of the new faith. Therefore, four epigrams record the conversion of the temple of Tyche into a tavern (9.180-183) and 10.90 seems to attack the doctrine of the Resurrection. When religion and politics go hand-in-hand, when religious conspiracies link up with palace intrigues, consequences are predictable: a man called Doroteo denounced him for his negative response to the new dogma, which caused him the loss of his teacher paid work. His annoyance was even greater with further sufferings, chiefly economic, and he had to sell his books (9.171 and 9.175) among other desperate decisions Forewarned about his radical bitterness against Church, it is just to make clear that he is not fully acquiescent with ancient gods and heroes. Thus, in 5.257 he questions Zeus’ ars amandi, in 9.377 refutes Tantalus’ possibility of thirst and hunger in Hades and 9.773 mischievously points out that Eros has been changed into a pan. The work begins with the selection, personal translation and comprehensive analysis of twenty two epigrams. Through such philological aid, we attempt to verify the frictions and the main perceptible factors in his poetic creation to justify his worldview, according to the pagan sentiment widespread in that time before the twilight already overwhelming of its ailing traditions

Teología, 2003, Tomo XL nº 082 (número completo)

Nota del Director. Novedades del 2004 -- Lo objetivo y lo subjetivo de la redención cristiana. Síntesis histórica y perspectiva actual / Leonardo Capelluti -- En torno a la beatificación de Isabel la Católica. Amor a la Iglesia. Pobreza / Melquíades Andrés Martín -- Razón teológica y tutela del embrión humano / Alejandro C. Llorente -- Religión y Ciencia en el pensamiento de Juan Pablo II: importancia y responsabilidad del diálogo para una cultura verdaderamente humana / Jorge Papanicolau -- La Iglesia en Buenos Aires durante la epidemia de fiebre amarilla de 1871 según el Diario de la epidemia Mardoqueo Navarro / Jorge Ignacio García Cuerva -- Ars artium: vigencia de la lógica clásica / Oscar H. Beltrán -- Crónica de la Facultad 2003

Fatigue crack-growth rate in ferrite martensite dual-phase steel

An empirical study is made on the fatigue crack growth rate in ferrite-martensite dual-phase (FMDP) steel. Particular attention is given to the effect of ferrite content in the range of 24.2% to 41.5% where good fatigue resistance was found at 33.8%. Variations in ferrite content did not affect the crack growth rate when plotted against the effective stress intensity factor range  which was assumed to follow a linear relation with the crack tip stress intensity factor range ΔK. A high  corresponds to uniformly distributed small size ferrite and martensite. No other appreciable correlation could be ralated to the microstructure morphology of the FMDP steel. The closure stress intensity factor , however, is affected by the ferrite content with  reaching a maximum value of 0.7. In general, crack growth followed the interphase between the martensite and ferrite.

Dividing the fatigue crack growth process into Stage I and II where the former would be highly sensitive to changes in ΔK and the latter would increase with ΔK depending on the  ratio. The same data when correlated with the strain energy density factor range ΔS showed negligible dependence on mean stress or R ratio for Stage I crack growth. A parameter α involving the ratio of ultimate stress to yield stress, percent reduction of area and R is introduced for Stage II crack growth so that the  data for different R would collapse onto a single curve with a narrow scatter band when plotted against αΔS.

Aportaciones a la escultura romanista en Gipuzkoa. Pedro de Goicoechea y el retablo mayor de San Miguel de Irura

[ES] En la iglesia de San Miguel Arcángel, en Irura, encontramos uno de los retablos más interesantes de finales del Renacimiento en Gipuzkoa. El altar posee relieves y tallas de estilo romanista de gran calidad, junto con una mazonería manierista. La obra fue realizada en las postrimerías del siglo XVI por los artistas Pedro de Goicoechea y Jerónimo de Larrea, miembros del taller de Tolosa, siguiendo las trazas dadas por escultor Juan de Goicoechea.

Redes de doadores e financiamento político em 2014: distinguindo doações para candidatos, partidos e comitês financeiros nas eleições presidenciais brasileiras

Aplica a metodologia de Análise de Redes Sociais (ARS) para caracterizar o financiamento de campanhas nacionais brasileiras a partir das redes constituídas pelos doadores. Os resultados mostram diferenças nas estratégias de arrecadação dos partidos e maior consistência ideológica na formação da rede de doadores partidários, frente aos doadores a candidatos e aos comitês partidários.

Isolation and characterization of proteins that bind to yeast origins of DNA replication

Yeast chromosomes contain sequences called ARSs which function as origins of replication in vitro and in vivo. We have carried out a systematic deletion analysis of ARS1, allowing us to define three functionally distinct domains, designated A, B, and C. Domain A is a sequence of 11 to 19bp, containing the core consensus element that is required for replication. The core consensus sequence, A/TTTTATPuTTTA/T, is conserved at all ARSs sequenced to date. A fragment containing only element A and 8 flanking nucleotides enables autonomous replication of centromeric plasmids. These plasmids replicate very inefficiently, suggesting that flanking sequences must be important for ARS function. Domain B also provides important sequences needed for efficient replication. Deletion of domain B drastically increases the doubling times of transformants and reduces plasmid stability. Domain B contains a potential consensus sequence conserved at some ARSs which overlaps a region of bent DNA. Mutational analysis suggests this bent DNA may be important for ARS function. Deletion of domain C has only a slight effect on replication of plasmids carrying those deletions.

We have identified a protein called ARS binding factor I (ABF-I) that binds to the HMR-E ARS and ARS1. We have purified this protein to homogeneity using conventional and oligonucleotide affinity chromatography. The protein has an apparent molecular weight of 135kDa and is present at about 700 molecules per diploid cell, based on the yield of purified protein and in situ antibody staining. DNaseI footprinting reveals that ABF-I binds sequence-specifically to an approximately 24bp sequence that overlaps element Bat ARSl. This same protein binds to and protects a similar size region at the HMR-E ARS.

We also find evidence for another ARS binding protein, ABF-III, based on DN asei footprint analysis and gel retardation assays. The protein protects approximately 22bp adjacent to the ABF-I site. There appears to be no interaction between ABF-I and ABF-III despite the proximity of their binding sites.

To address the function of ABF-I in DNA replication, we have cloned the ABF-I gene using rabbit polyclonal anti-sera and murine monoclonal antibodies against ABF-I to screen a λgt11 expression library. Four EcoRI restriction fragments were isolated which encoded proteins that were recognized by both polyclonal and monoclonal antibodies. A gene disruption can now be constructed to determine the in vivo function of ABF-I.

Donati Artis Grammaticae Index et Concordantia

615 p.