The role of MAR elements in DNA recombination

The untargeted integration of foreign DNA into the mammalian cell genome, extensively used in gene therapy and biotechnology, remains an incompletely understood process. It is believed to be based on cellular DNA double strand break (DSB) repair machinery and to involve two major steps: i) the formation of long gene arrays (concatemers), and ii) recombination of the resulting concatemer with the genome. The main DSB repair pathways in eukaryotes include non-homologous end-joining (NHEJ), homologous recombination (HR), and microhomology-mediated end-joining (MMEJ). However, it is still not clear, which of these pathways are responsible for transgene integration. Here, we show that NHEJ is not the primary pathway used by mammalian cells in the transgene integration process, while the components of the HR pathway seem to be important for genomic integration but not concatemerization. Instead, concatemer formation appears to be mediated by a subset of the MMEJ pathway, termed synthesis-dependent MMEJ (SD-MMEJ). This mechanism also seems to be preferentially used for plasmid integration into the genome, as confirmed by the analysis of plasmid-to-genome junction sequences, which were found to display an SD-MMEJ pattern. Therefore, we propose the existence of two distinct SD-MMEJ subpathways, relying on different subsets of enzymes. One of these mechanisms appears to be responsible for concatemerization, while the other mechanism, partially dependent in HR enzymes, seems to mediate recombination with the genome. Previous studies performed by our group suggested that matrix attachment regions (MARs), which are epigenetic regulatory DNA elements that participate in the formation of chromatin boundaries and augment transcription, may mediate increased plasmid integration into the genome of CHO cells by stimulating DNA recombination. In the present work, we demonstrate that MAR-mediated plasmid integration results from the enhanced SD-MMEJ pathway. Analysis of transgene integration loci and junction DNA sequences validated the prevalent use of this pathway by the MAR elements to target plasmid DNA into gene-rich areas of the CHO genome. We propose that this finding should in the future help to engineer cells for improved recombinant protein production. In addition to investigating the process of transgene integration, we designed recombination assays to better characterize the components of the MMEJ and SD-MMEJ pathways. We also used CHO cells expressing cycle-sensitive reporter genes to demonstrate a potential role of HR proteins in the cell cycle regulation.

El medi marí del cap de Creus i el golf de Roses

El medi marí de cap de Creus i golf de Roses té unes característiques ecològiques, geològiques i oceanogràfiques excepcionals que han permès el desenvolupament turístic i pesquer de les viles costaneres adjacents, Aquest article sintetitza els principals valors ecològics i econòmics de l'àmbit marí d'aquests dos espais inclosos dins la candidatura de Patrimoni Mundial de la UNESCO

Entrevista a Mar Cruz Piñol

Mar Cruz Piñol es profesora titular de Lengua Española en la Facultad de Filología de la Universidad de Barcelona, donde desde 1995 ha impartido, entre otras, las asignaturas"Cuestiones del español como L2","Lingüística de corpus","Bases de datos para el estudio de la lengua española" y"Metodología de la investigación filológi- ca".

Espectrofotometría de absorción atómica con tubo en la llama: aplicación en la determinación total de cadmio, plomo y zinc en aguas frescas, agua de mar y sedimentos marinos

Thermospray flame furnace Atomic Absorption Spectrometry (TS-FF-AAS) was used for the total determination of Cd, Pb and Zn in fresh water and seawater samples at µg L-1 levels, and in marine sediment samples at µg g-1 levels. Using a sample loop of 50 µL and a peristaltic pump the samples were transported into the metallic tube placed over an air/acetylene flame, through a ceramic capillary (o.d. = 3.2 mm) containing two parallel internal orifices (i.d = 0.5 mm). The detection limits determined for Cd, Pb and Zn using a synthetic water matrix (2.5% m/v NaCl, 0.5% m/v MgCl2 and 0.8% m/v CaCl2) were 0.32 µg L-1; 2.6 µg L-1 and 0.21 µg L-1 respectively. The methodology by TS-FF-AAS was validated by determination of Cd, Pb and Zn in certified reference materials of water and marine sediment, and the t-test for differences between means was applied. No statistically significant differences were established in fresh water and seawater (p>0.05), whereas differences became apparent in marine sediment (p<0.03).

Estimativa do fluxo de amônia na interface ar-mar na Baía de Guanabara: estudo preliminar

In the begining of April 2004, concentrations of NHx (NH3 + NH4+) were measured in surface waters of the Guanabara Bay. Concentrations varied from 2 to 143 mmol L-1. Ammonia exchange at the air-sea interface was quantified using a numerical model. No measurement of NH3 concentration in air (c air) was performed. Thus, calculations of NH3 flux were based on the assumptions of c air = 1 and 5 µg m-3. Fluxes were predominantly from the water to the atmosphere and varied from -20 to almost 3500 µg N m-2 h-1.

Informe d'Autors UOC a ISI Web of Knowledge. Març 2015

UOC

Navegar y rezar: devoción y piedad de las gentes de mar barcelonesas (siglos XIV y XV)

Sobre la devoció i la pietat religiosa de la gent de mar de Barcelona dels segles XIV i XV

Fluxos de óxido nitroso na interface ar-mar na Baía de Guanabara

In Surface water concentrations of N2O were measured at 37 stations in Guanabara Bay and fluxes estimated across the air-sea interface. Concentrations averaged 8.2 ± 2.2 nmol L-1 and 90% of the stations showed supersaturation averaging 33%. N2O fluxes were estimated using a two-film model which is given by the product of the concentration difference across the film and the gas transfer coefficient (k w). Two parametrizations of k w were used which provided average fluxes of 0.3 and 3.0 µg N m-2 h-1. Flux measurements using floating chambers (not reported here) seem to agree with the upper limit of these estimates.

Preconcentración y atomización con tubo en la llama para la determinación de Cadmio en agua de mar

A flow injection on-line pre-concentration system coupled to thermospray flame furnace atomic absorption spectrometry (TS-FF-AAS) for cadmium determination at sub μg L-1 levels in seawater samples was developed. The on-line system was evaluated by analysing cadmium containing in a synthetic seawater matrix (2.5% m/v NaCl, 0.5% m/v MgCl2 and 0.8% m/v CaCl2). A sample volume of 2 mL allows determining Cd with a detection limits of 30 ng L-1 (3* σblank/slope), pre-concentration factor of 34 and repeatability of 1,8% (calculated as RSD, N=8 and containing 200 ng L-1 of Cd ).

Colonização natural por espécies nativas e exóticas das encostas degradadas da Serra do Mar

Os deslizamentos são eventos inerentes à dinâmica da Serra do Mar, nos trechos mais declivosos. Contudo, devido a alguns fatores de intervenção, como a poluição atmosférica, ocupação da base dos morros e outros, a incidência desses eventos tem aumentado. Diante disso, este estudo objetivou avaliar a dinâmica de colonização de áreas de encosta degradadas, como forma de subsidiar ações de recuperação. Para tanto, foram adotadas duas áreas experimentais, localizadas em trechos da serra do Mar próximos à Baixada Santista, onde foram lançadas parcelas de 1 m² e obtidos dados fitossociológicos. Em seguida, as áreas foram limpas e os mesmos parâmetros da avaliação inicial, registrados, mensalmente, ao longo de um ano. Foram encontradas 20 espécies, pertencentes a 14 famílias botânicas, em sua maioria espécies exóticas ou nativas não regionais.

Mar del Nort

Julkaisussa: Het vyfde deel des Grooten Atlas, vervattende de water-weerelt. Vol. V

Il disegno della geografia moderna de tutta la parte dell'Africa i confini della quale stanno in questo modo, da ponente il mar'oceano computate l'isole di Capo Verde, et le Canarie da Tramontana il Stretto Gibelterra, et il Mare Meditteraneo ...

Kartta kuuluu A. E. Nordenskiöldin kokoelmaan

O mar e a alma: metáforas marinhas em território alemão

O artigo explora algumas metáforas marinhas que surgem no pensamento alemão de Leibniz a Goethe, com o intuito de indicar como se desenvolvem certos temas de estética, tais como a noção de alma, de linguagem, de criação artística e de relação dialética entre forma e conteúdo. Passando por autores como Leibniz, Winckelmann, Herder, Goethe e Kant, pretende-se mostrar como, por meio desse desenvolvimento, se constitui uma visão de homem mais ampliada, que não se define mais somente pelo entendimento, mas envolve elementos inconscientes e afetivos.