DNA double-strand breaks measured by the non-denaturing filter elution technique following X-ray and restriction endonuclease treatment

The non-denaturing filter elution technique of Bradley and Kohn (1979) is widely used as an assay for DNA double-strand breaks (dsb) in mammalian cells. Results characteristically obtained with this assay following exposure of cells to ionising radiation, namely that of non-linear induction of dsb and rapid biphasic repair, are not in agreement with those of the neutral velocity sedimentation technique where the biophysical basis of measurement is understood. The discrepancy in the results obtained with these two techniques with regard to both Induction and repair of dsb has led to a controversy. In particular concerning the manner in which the data obtained with the neutral elution technique should be interpreted (AhnstrSm's Comment on Radford 1985; Hutchinson 1989). The aim of this project was therefore to attempt to test the assumed specificity of the non-denaturing filter elution technique as an assay for dsb. Optimization of the lysis and eluting conditions was followed by detailed X-ray dose-response and repair experiments with the CHO K1 cell line. A comparative study was performed using the xrs 5 cell line, a radiosensitive mutant of the CHO K1 line chosen for its characteristic marked deficiency in dsb repair, yet normal ability to rejoin single-strand breaks (Kemp et al 1984; Costa and Bryant 1988). Previous reports by Bryant and Blocher (1982), and Iliakis and Bryant (1983) revealed that the DNA synthesis Inhibitors ara A and ara C strongly inhibit dsb repair as assayed by neutral velocity sedimentation. I thus adopted an experimental strategy In which the effect of ara A and ara C on putative dsb repair was examined using the non- denaturing filter elution assay. Only limited inhibition of dsb repair by these nucleoside analogues was observed with the non-denaturing filter elution technique tn contrast to the complete inhibition of dsb repair as measured by neutral velocity sedimentation for the same concentrations of DNA synthesis inhibitor (Bryant and Blocher 1982; Iliakis and Bryant 1983). These results suggest that the two above mentioned techniques are detecting disparate types of dsb, as manifested by the differential requirement of the repair mechanism of these breaks for DNA polymerization. A further approach was the introduction of restriction endonucleases (RE) into mammalian cells by electroporation, to induce dsb in the absence of other types of lesions. The observed increase in the rate of elution of the DNA of RE-treated cells substantiates the ability of the non-denaturing filter elution assay to detect cellular dsb. Surprisingly Pvu II was found to remain active inside the cell for up to 24 h, and the continual incision of the DNA by the enzyme thwarted the possibility of monitoring the repair of these dsb. A noteworthy result was the relative inability of RE which generate cohesive-ended dsb (e.g. Bam HI and Eco R1) to Induced measurable numbers of dsb as compared with the blunt-end cutter Pvu II. A hypothesis of a competition between the induction of dsb by RE and the subsequent repair is offered as explanation, where the repair of cohesive-ended dsb is assumed to take place at a higher rate than that of blunt-ended dsb. A comparative study using the xrs 5 mutant cell line, known to be deficient in dsb repair, revealed enhanced levels of RE-induced dsb which would support the notion that the levels of dsb reflect a competition between RE-incision and dsb repair. In summary, this study validates the measurement of dsb by the non-denaturing filter elution method and provides new evidence for the mode of induction of dsb by RE which has not been hitherto possible. Finally, the work indicates the way in which cells handle different types of dsb which may be similar to the manner In which the variety of dsb induced by ionising radiation are dealt with

Towards a design observatory: crafting a distributed approach

As Design is gaining traction globally different observation models were developed to map the Design landscape. However, the application of these models in contexts with a slower maturation of Design culture and no institutional Design infrastructure can be difficult. With this challenge in mind, this paper presents a new distributed approach to support Design observation. It applies it to a case within the DesignOBS project - a project aiming to identify, map and interpret the Portuguese design landscape - and develops an online platform with the aim to create more efficient and engaging representations of Design practices and realities/contexts to multiple publics, including policymakers. The approach can benefit countries in a similar situation as Portugal, leveraging the participation of design actor- networks, encourage local initiatives, map the evolving Design landscape in collective manner, and hopefully constitute the supporting, resilient backbone to develop national Design policy.Project Design Obs. Towards a design Observatory in Portugal: models, instruments, representation and strategies supported by Lisbon Regional Operational Program (LISBOA 2020) and the Competitiveness and Internationalization Operational Program (POCI-01-0145-FEDER-032445), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and FCT – Foundation for Science and Technology.publishe

Design infrastructures: proposing alternative strategies for countries with a lower maturation of design culture

In Europe, Design Centers and Associations are considered as key infrastructures to efficiently promote and represent the discipline. However, in some countries with a lower maturation of Design culture – such as the case of Portugal - there are no official actors fully dedicated to these activities. Previous research indicated Design schools as a potential alternative infrastructure to promote and represent Design, but further research is needed to understand what they can learn/adapt from the activities currently undertaken by Centers and Associations. This paper maps the European landscape looking in particular at BEDA (Bureaux of European Design Association) members. Based on these insights and recent infrastructuring literature, it develops adapted strategies for observation activities to test with Design schools in the Portuguese territory. The results obtained are a first step to bring countries with lower maturation of design culture under the EU Design Ecosystem radar.Project Design Obs. Towards a design Observatory in Portugal: models, instruments, representation and strategies supported by Lisbon Regional Operational Program (LISBOA 2020) and the Competitiveness and Internationalization Operational Program (POCI-01-0145-FEDER-032445), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and FCT – Foundation for Science and Technology.in publicatio

Uma perspetiva sobre os objetos de design concebidos a propósito da pandemia de COVID-19

A presente dissertação visa a discussão do tema “A arte a técnica no design contemporâneo”. A arte e a técnica quando caminham juntas formam a ponte para o design. Com base nisto, há a perceção de como a arte e a técnica no design interagem numa situação de emergência, como a SARS-COV-2. A parte inicial é uma abordagem histórica desde o século XIX ao século XX, devido a ter sido nestes séculos que começou a surgir a dicotomia da indústria e da arte e, por consequência, a sua simbiose, que origina o design. Na atualidade há a compreensão de como o design interage com a arte e a técnica. Se por um lado há o design na sua dimensão mais artística e daqui surge o design crítico e especulativo, por outro lado temos a dimensão mais técnica no design. Faz-se assim uma análise de como arte e técnica dão a valorização ao design, tornando-o com objetivos concretos e tendo ele um papel fulcral na sociedade. O design é feito para pessoas/utilizadores e com a função/tentativa de resolver algo com base na sociedade. E é a concordância destas duas dimensões, tanto a dimensão mais técnica, como a dimensão mais artística que dá o surgimento de um design mais consciente. Um design consciente que tem por base a realização de algo funcional, necessário, durável e sustentável. Com base nisto, questionamos como é que estas duas dimensões atuam no design realizado num estado de emergência, mais precisamente na pandemia Covid-19 e como os objetos caracterizam esta situação tão específica.The aim of this dissertation is to discuss the theme the art and the technique in the contemporary design. When together, art and technique lead to design. Based on this, there is the awareness of how art and technique interact in an emergency situation, such as SARS-COV-2. The initial part is a historical approach from the 19th to the 20th century, since it was then that the dichotomy between industry and art began to emerge, and, consequently, its symbiosis which has originated design. Nowadays, there is an understanding of how design interacts with the art and the technique. On the one hand, design has a more artistic dimension and so the critical and speculative design has appeared; on the other hand, there is a more technical dimension of design. This way, it is analysed how art and technique value design, making it more objective and having a central role in society. Design is made for people/users and has the function/attempt to solve something based on society. Therefore, it is the relationship between these two dimensions, both the more technical and the more artistic, that permits a more conscious design. A conscious design based on making something functional, necessary, durable and sustainable. Based on this, it is questioned the way these two dimensions act in the design during a state of emergency, more precisely in the Covid-19 pandemic, and how the objects characterise such a specific situation

Acogimiento familiar: caracterización de un programa

Este artigo objetiva caracterizar os principais atores envolvidos em um programa de Acolhimento Familiar e os acolhimentos realizados. Esse programa efetuou 314 acolhimentos entre 1998 e 2007. Neste período, a maioria dos acolhidos eram meninos (51,7%), afrodescendentes (55,3%), de até seis anos (58,7%). As famílias acolhedoras eram constituídas por casais (70,4%), com filhos (84,1%); faixa etária entre 30 e 49 anos; bom nível de escolarização; profissões autônomas; rendas diversificadas e motivações solidárias para acolher. Com relação às famílias de origem não foram encontradas informações. Como principal causa dos acolhimentos foi citada a negligência (33,7%). Sua duração foi de um a seis meses em 46,65% dos casos. Quanto aos encaminhamentos pós-acolhimento, 34,2% das crianças foram adotadas e 33,8%, reintegradas às suas famílias. Caracterizar um programa oferece subsídios para a redefinição de práticas e auxilia na formulação de ações de intervenção em situações complexas como o acolhimento familiar.This paper aims to describe the main actors involved in a Foster Care program and its processes. The program had done 314 fosterings from 1998 to 2007. During this period, most foster children were boys (51,7%), afrodescendants (55,3%), aged up to 6 years old (58,7%). Foster families were formed by couples (70, 4%), with other children (84,1%); aged from 30 to 49 years old; good education level; autonomous professions; diversified incomes and solidarity motivation to foster. No registered information about the biological families was found. The principal reason referred to for fostering was negligence (33,7%). The child remained in foster care from 1 to 6 months in 46,65% cases. Regarding the outcome of the fostering process, 34,2% children were adopted and 33,8% were reintegrated into their families. Describing a program helps to redefine practices and to discuss measures of intervention for the area.El artículo objetiva conocer los principales envueltos en un programa de Acogimiento Familiar y los acogimientos realizados. Ese programa efectuó 314 acogimientos entre 1998 y 2007. En ese período, la mayoría de los acogidos eran niños (51,7%), afrodescendientes (55,3%), hasta 6 anõs (58,7%). Las familias acogedoras eran constituidas por parejas (70,4%), con hijos (84,1%); edad entre 30 y 49 años; buen nivel de escolaridad; profesiones autónomas; ingresos diversificados y motivaciones solidarias para acoger. Sobre las familias de origen no fueron encontradas informaciones. La principal causa de los acogimientos fue la negligencia (33,7%), y su duración de 1 a 6 meses in 46,65% de los casos. Sobre las derivaciones de los acogimientos, 34,2% de los niños fueran adoptados y 33,8%, regresaron a sus familias. Describir un programa ofrece subsidios para la redefinición de prácticas y puede auxiliar a repensar acciones de intervención en el área

PORTUGAL WAVE 3. Pre-election Study

This study is part of the MAPLE Project, ERC – European Research Council Grant, 682125, which aims to study the Politicisation of the EU before and after the Eurozone Crisis in Belgium, Germany, Greece, Ireland, Portugal and Spain. In each of these countries, an online panel is to be carried out just before and just after the legislative elections. This report pertains to the pre-election panel of Portugal legislative elections 2019 held on 6 October. Our questionnaire seeks to model the political context of political choices, and to understand the importance that European attitudes may have in voting behaviour. In Portugal, we have partnered with Netquest. We present in this report a number of political attitudes according to stated partisanship in Portugal. We are interested in the way in which partisan preferences are related to political attitudes, including national issues as well as those pertaining to the EU. The target population is the general voting population aged >18 years old using the Census 2011 data. The sample size is 1540. The fieldwork was done using the Netquest online panel and took place between 19/08/2019 and 19/09/2019. The sample followed and fulfilled a socio demographic matrix which crossed three quotas: gender (male, female); age (18-34; 35-54; 55+); education (less than secondary; secondary; more than secondary). The resulting dataset was weighted according to a weight combining gender (male, female), age (18-34; 35-54; 55+), and education (up to secondary; secondary; postsecondary). Our sample included weights above 2 for less than 10% of the sample. To measure partisanship we used the following questions: the respondent was asked if s/he felt close to a party. To those who replied no in the first question, they were asked again, namely if they felt closer to one party than others. Only those who repeated no in this question too were coded as “without party id”. All others were coded with their stated party identification. In the results we show all parties that 1% or more of respondents identify with. Throughout the report, we will only highlight differences that are substantive, meaning that the differences are statistically significant.info:eu-repo/semantics/publishedVersio