Addition of a histone deacetylase inhibitor increases recombinant protein expression in Medicago truncatula cell cultures

Plant cell cultures are an attractive platform for the production of recombinant proteins. A major drawback, hindering the establishment of plant cell suspensions as an industrial platform, is the low product yield obtained thus far. Histone acetylation is associated with increased transcription levels, therefore it is expected that the use of histone deacetylase inhibitors would result in an increase in mRNA and protein levels. Here, this hypothesis was tested by adding a histone deacetylase inhibitor, suberanilohydroxamic acid (SAHA), to a cell line of the model legume Medicago truncatula expressing a recombinant human protein. Histone deacetylase inhibition by SAHA and histone acetylation levels were studied, and the effect of SAHA on gene expression and recombinant protein levels was assessed by digital PCR. SAHA addition effectively inhibited histone deacetylase activity resulting in increased histone acetylation. Higher levels of transgene expression and accumulation of the associated protein were observed. This is the first report describing histone deacetylase inhibitors as inducers of recombinant protein expression in plant cell suspensions as well as the use of digital PCR in these biological systems. This study paves the way for employing epigenetic strategies to improve the final yields of recombinant proteins produced by plant cell cultures.publishersversionpublishe

Avaliação de efeitos ecológicos da interdição da pesca lúdica no litoral rochoso alentejano

A pesca lúdica no Parque Natural do Sudoeste Alentejano e Costa Vicentina (PNSACV) foi sujeita em 2009 a restrições específicas, que incluem a sua interdição na zona da Ilha do Pessegueiro, onde posteriormente foi criada uma área de proteção. Este estudo teve por objetivo avaliar os efeitos ecológicos de 2 anos (2009 - 2011) de proteção marinha, propor metodologias de monitorização e sugerir melhorias na gestão das pescas lúdica e comercial. Averiguaram-se os efeitos ecológicos da proteção na abundância, distribuição e dimensão de organismos intertidais (lapas e burriés) habitualmente explorados por pescadores lúdicos no litoral rochoso da área referida, e na restante comunidade de organismos macroepibentónicos. Comparou-se a área protegida com áreas de controlo não protegidas. As análises de variância não revelaram efeitos da proteção ao nível da área nem do local na abundância e tamanho de lapas e burriés. Foi detetada variabilidade à escala do local. Comparando com dados anteriores a abundância e o tamanho de lapas diminuiu mas a abundância de lapas grandes manteve-se. O tempo de recuperação insuficiente, os efeitos predatórios de outros consumidores de moluscos e a deficiente eficácia da proteção, são possíveis causas para os resultados encontrados. Considera-se essencial a resolução do conflito social. Propõe-se a governância colaborativa como estratégia de gestão da área marinha protegida do PNSACV; ABSTRACT: Recreational fishing at Parque Natural do Sudoeste Alentejano e Costa Vicentina (PNSACV) was limited by legal restrictions, including fishing ban in Ilha do Pessegueiro and continental adjacent area. All the area was later classified as protected. The aim of this study was to evaluate the ecological effects of a 2 year period (2009-2011) of marine protection, to propose monitoring methodologies and suggest management improvements for recreational and commercial fishing. Ecological effects of the stated protection were studied on intertidal organisms (limpets and sea snails) that use to be explored by recreational fishers on the rocky shores of the cited area, and on the community of macro-epibenthic organisms. The protected area was compared with non protected control areas. Variance analysis didn’t show effects of protection at both area and local levels in the abundance and size of limpets and sea snails. Variability at local scale was detected. Compared with previous data the abundance and size of limpets decreased but the abundance of large limpets maintained. Insufficient recovery time, predation by other mollusk consumers and deficient regulatory enforcement are possible causes for the obtained results. Social conflict resolution is essential. Collaborative governance is suggested as a management strategy for the marine protected area in PNSACV

Comunicação de ciência: práticas e representações entre investigadores

Mestrado em Comunicação em CiênciaEste trabalho debruça-se sobre a temática da comunicação de ciência, e mais especificamente sobre as práticas e as representações dos investigadores do IBMC. Partindo de uma revisão da literatura sobre o tema, identificaram-se os níveis de participação em actividades de comunicação de ciência dirigidas aos media, público escolar e público geral, em função de características biográficas; analisaram-se as percepções dos investigadores sobre diversos aspectos da comunicação de ciência: atitudes, significados e condicionantes da sua participação. Analisaram-se ainda as representações de investigadores com práticas distintas para avaliar a relação entre os níveis de participação e as respectivas representações. Desta investigação resultam três conclusões principais: os investigadores do IBMC, na sua grande maioria participam em actividades de comunicação de ciência, investigadores com níveis de participação elevados expressam atitudes mais positivas do que aqueles com menores níveis de participação, e a instituição de investigação tem um papel determinante nos níveis de participação dos seus investigadores. ABSTRACT: This study is developed within the field of science communication. It focuses on the practices and representations about activities of science communication of researchers at IBMC. Following a literature review we have identified the levels of participation on science communication activities aimed to the media, schools and general public, considering the researcher’s biographic profile. We have analysed the attitude, the perceptions and circumstances in which science communication activities take place. We have also studied the conceptions of researchers with distinct practices in order to assess the relationship between the level of participation and the corresponding conceptions about science communication. Among the main findings we highlight three: IBMC researcher’s revealed high levels of participation in science communication activities, researchers with higher levels of participation express a more positive attitude towards science communication than the researchers with lower participation levels and the institution plays an important role in researcher’s level of participation

In situ methods to localize transgenes and transcripts in interphase nuclei: a tool for transgenic plant research

Genetic engineering of commercially important crops has become routine in many laboratories. However, the inability to predict where a transgene will integrate and to efficiently select plants with stable levels of transgenic expression remains a limitation of this technology. Fluorescence in situ hybridization (FISH) is a powerful technique that can be used to visualize transgene integration sites and provide a better understanding of transgene behavior. Studies using FISH to characterize transgene integration have focused primarily on metaphase chromosomes, because the number and position of integration sites on the chromosomes are more easily determined at this stage. However gene (and transgene) expression occurs mainly during interphase. In order to accurately predict the activity of a transgene, it is critical to understand its location and dynamics in the three-dimensional interphase nucleus. We and others have developed in situ methods to visualize transgenes (including single copy genes) and their transcripts during interphase from different tissues and plant species. These techniques reduce the time necessary for characterization of transgene integration by eliminating the need for time-consuming segregation analysis, and extend characterization to the interphase nucleus, thus increasing the likelihood of accurate prediction of transgene activity. Furthermore, this approach is useful for studying nuclear organization and the dynamics of genes and chromatin

Neoplasia Maligna da Próstata e Fertilidade: Revisão das Implicações das Diferentes Opções Terapêuticas

Introduction: Prostate cancer is the second most common neoplasm in men, and its incidence is higher in the group aged 50-69 years. It is estimated that about 5% of cases are diagnosed in men under the age of 50. Given the current demographic developments, the desire for paternity is often postponed. Thus, it is relevant that the urologist can explain the different repercussions that the existing therapies for malignant prostate cancer have on male fertility. The purpose of this systematic review is to compare the repercussions that the diverse therapeutic options used in prostate cancer have on male fertility. Methods: a qualitative review of the scientific literature published between 1987-2019, according to the ENTREG guidelines and PRISMA methodology, through PubMed database. All articles written in English and covering the topics described were consi- dered, including randomized controlled studies, meta-analyses and systematic reviews. All the abstracts of the publications were read by two authors, with the consequent selection of those that met the defined criteria, after evaluating its quality using the CASP qualitative checklist. In case of divergent opinions, a third urologist was consulted to decide on inclusion. Results: According to the defined criteria, 436 references were identified and 10 publications were included as they refer directly to the defined objective. Despite the few published studies, there is evidence that active surveillance alone has no implications for fertility. Although there are no comparative studies, according to the currently available data, focal therapy and brachytherapy seem to be the therapeutic options with the least likelihood of permanent impairment of the capacity for natural fertilization as opposed to radical prostatectomy, radiotherapy, hormonotherapy and chemotherapy. Conclusion: Nowadays, with increased survival after prostate cancer treatment, it is relevant to evaluate the repercussions on fertility of the existing therapeutic options for prostate cancer, so that patients can make an informed decision and, if necessary, resort to gametes preservation measures prior to therapeutic intervention. Although there are no comparative studies, focal therapy and brachytherapy appear to have less effect on male fertility. Active surveillance is currently the only option with no consequences on fertility.Introdução: A neoplasia maligna da próstata é a segunda neoplasia mais comum nos homens, e a sua incidência é mais elevada no grupo com 50-69 anos de idade. Estima-se que cerca de 5% dos casos sejam diagnosticados em homens com idade inferior a 50 anos. Dada a evolução demográfica actual, o desejo de paternidade é frequentemente adiado. Assim, é relevante que o urologista possa explicar as diferentes repercussões que as terapêuticas existentes para a neoplasia maligna da próstata têm na fertilidade masculina. O objectivo desta revisão é comparar o impacto que as diferentes opções terapêuticas existentes na neoplasia maligna da próstata têm na fertilidade masculina. Métodos: Realização de revisão qualitativa da literatura publicada entre 1987-2019, de acordo com as orientações ENTREG e metodologia PRISMA, utilizando a base de dados PubMed. Foram considerados elegíveis todos os artigos sobre este tema escritos em inglês, incluindo estudos randomizados controlados, meta-análises e revisões sistemáticas. Todos os resumos foram lidos por dois autores com subsequente selecção daqueles que cumprissem os critérios definidos, após avaliação de qualidade utilizando a checklist qualitativa CASP. Em caso de discordância, um terceiro urologista foi consultado. Resultados: De acordo com os critérios definidos, foram identificadas 436 referências e 10 publicações foram incluídas por se referirem directamente ao objectivo definido. Apesar da escassez de literatura publicada, é possível concluir que a vigilância activa não tem implicações na fertilidade. De acordo com os dados disponíveis, a terapêutica focal e a braquiterapia são as opções terapêuticas com menor probabilidade de provocar incapacidade permanente na fertilização por métodos naturais, em oposição a prostatectomia radical, radioterapia, hormonoterapia e quimioterapia. Conclusão: Actualmente, com o aumento da sobrevivência após tratamento de cancro da próstata, é necessário avaliar as repercussões que as diferentes opções terapêuticas têm na fertilidade, de forma que os doentes possam fazer uma decisão informada e, se necessário, recorrer a estratégias de preservação de gâmetas. Apesar de não existirem estudos comparativos, a terapêutica focal e a braquiterapia parecem ter menor efeito deletério na fertilidade masculina. A vigilância activa é actualmente a única opção sem consequências na fertilidade

Evaluation of ecological effects of recreational fishing ban in the Alentejo rocky shores

Mestrado em Gestão e Conservação de Recursos Naturais - Instituto Superior de Agronomia / Universidade de ÉvoraRecreational fishing at Parque Natural do Sudoeste Alentejano e Costa Vicentina (PNSACV) was limited by legal restrictions, including fishing ban in Ilha do Pessegueiro and continental adjacent area. All the area was later classified as protected. The aim of this study was to evaluate the ecological effects of a 2 year period (2009-2011) of marine protection, to propose monitoring methodologies and suggest management improvements for recreational and commercial fishing. Ecological effects of the stated protection were studied on intertidal organisms (limpets and sea snails) that use to be explored by recreational fishers on the rocky shores of the cited area, and on the community of macro-epibenthic organisms. The protected area was compared with non protected control areas. Variance analysis didn’t show effects of protection at both area and local levels in the abundance and size of limpets and sea snails. Variability at local scale was detected. Compared with previous data the abundance and size of limpets decreased but the abundance of large limpets maintained. Insufficient recovery time, predation by other mollusk consumers and deficient regulatory enforcement are possible causes for the obtained results. Social conflict resolution is essential. Collaborative governance is suggested as a management strategy for the marine protected area in PNSACV

Characterization of Medicago truncatula cell suspension cultures producing valuable recombinant proteins

First paragraph (this article has no abstract) Nowadays, the use of plants for large-scale production of recombinant proteins is gaining wider acceptance because of their many practical, economic and safety advantages, compared with traditional microbial and animal production systems.Peer reviewe