Production of transgenic Hypericum perforatum plants via particle bombardment-mediated transformation of novel organogenic cell suspension cultures

We have developed particle bombardment-mediated transformation procedure for Hypericum perforatum L. (St. John’s wort), an important medicinal species that remains highly recalcitrant towards Agrobacterium-mediated transformation. Among the major transformation techniques evaluated in the present study (Agrobacterium tumefaciens-, A. rhizogenes- and biolistics-mediated), particle-bombardment-mediated gene transfer was found to be the most successful one. GUS positive cells were obtained from organogenic nodules bombarded with the plasmid vector pCAMBIA1301 encoding an intron-containing b-glucuronidase (gusA) and hygromycin phosphotransferase (hpt) genes. After 3 months of continuous selection of bombarded nodules with 20 mg l 1 hygromycin, transgenic hygromycin-resistant callus cultures and subsequently transgenic plants were produced. PCR analysis of DNA isolated from GUS positive plants showed the presence of both gusA and hpt genes. Southern blot analysis confirmed the transgene integration and revealed diverse copy numbers and insertion sites. The data presented here demonstrate for the first time H. perforatum can be efficiently transformed via particle bombardment of organogenic cell suspension. Our results open the possibility of using particle bombardment-mediated transformation to elucidate biosynthetic pathways and to improve secondary metabolite production in H. perforatum.Fundação para a Ciência e a Tecnologia (FCT) - POCTI/AGR/40283/2001, SFRH/BPD/ 17102/200

Transgenic hypericum perforatum

Plant transformation is an important tool with many applications in modern plant biology. Although this technique is primarily used to produce superior crop varieties, it is also being utilized to answer basic questions concerning gene function and regulation in contemporary functional genomics research. In our laboratory, we have established a transformation system for Hypericum perforatum . This protocol involves the transfer of foreign DNA into H. perforatum organogenic nodule explants via particlebombardment and the regeneration of shoots from the explants under selection pressure. We have successfully used this method to express ß-glucuronidase and hygromycin phosphotransferase genes in H. perforatum . Molecular analyses of putative phenotypically normal transgenic plants show stable integration of the transgenes into the plant nuclear genome. Here we describe the procedure for the transformation of H. perforatum .Fundação para a Ciência e a Tecnologia (FCT

Challenges for modern wine production in dry areas: dedicated indicators to preview wastewater flows

Wine production is an important socio-economic activity in Mediterranean countries. This study is focused on wine production under warm and dry climate conditions in south Portugal, in two major wine-producing regions (Tejo and Alentejo), characterized by small to medium sized wineries. Vineyards have been expanding in this region of Portugal, where about 50–70% of the vineyards are irrigated, increasing regional water demand. The aim of this study is to propose an integrative approach for wine production, where a simple calculation model has been developed and validated to preview water consumption and wastewater production, as functions of winemaking periods and type of processed grapes. Results revealed a global ratio of 2.2 ±0.45 and 2.1 ±0.17 Lwater/Lwine. Concerning dedicated indicators, 60–75% of the wastewater was produced during Period I and the red wine production represented a 50–64% increase in water consumption. This tool will enable winemakers to calculate Global and Dedicated Indicators, based on their own parameters, which provide information on flow volumes and peak flows. In this context, it will be possible to identify improvements for wastewater treatment and management towards water reuse as a promising solution for the wine sector in the framework of the circular economyinfo:eu-repo/semantics/publishedVersio

Wine production wastes, valorization, and perspectives

Zlatina Genisheva wishes to thank you for the financial support to the project Mission2GG—To Grow Green, by PRR—Plano de Recuperação e Resiliência, enquadrado na Missão Interface.The wine sector generates high quantities of residues that are still poorly exploited as feedstock. Normally, these wastes are directly discarded into the fields or burned, thus causing environmental problems. Wine production wastes, like vine pruning and grape pomace, are available at relatively low prices and are onsidered prime materials for biochemical conversion into added-value products. In this context, the reutilization of these wastes is very important not only for minimizing environmental impact but also for obtaining higher profitability. The main objective of the present chapter is to address what are the possible reutilizations and valorizations of these wastes

Growing evidence supporting the use of mesenchymal stem cell therapies in multiple sclerosis: a systematic review

Background: Multiple sclerosis (MS) typically arises in early/middle adulthood and is characterized by a progressive disability of the central nervous system (CNS). Currently approved therapies do not promote tissue repair or stop disease progression. Emerging data demonstrate that stem cells present a great potential in regenerative medicine and, consequently, have also been widely investigated as a potential treatment for MS. Therefore, the aim of this study was to conduct a systematic review to inquire into the safety, tolerability, and efficacy of mesenchymal stem cells (MSCs) therapies in MS. Methods: Three electronic databases (Web of Science, PubMed, and Cochrane) were searched from April until June 2019. Clinical trials or case reports with information related to the effects of MSC therapies in MS patients were considered for this review. Results: 10 manuscripts were selected, namely 7 uncontrolled clinical trials, 2 randomized controlled clinical trials, and 1 case report. The overall quality of the studies was considered good. Besides minor adverse events (AEs), it was reported one case of encephalopathy with seizures and two cases of iatrogenic meningitis, which were not related to the treatment, but with the administration route. The analyses of the expanded disability status scale (EDSS) in the uncontrolled clinical trials demonstrated that 48 patients improved, 39 maintained and 16 worsened their clinical condition. Regarding the randomized studies, one did not show statistically significant variations in the mean EDSS score and in the other the mean EDSS score was statistically significantly lower for the experimental group. The case report also showed an improvement in the EDSS score. Conclusions: MSCs transplantation proved to be a safe and tolerable therapy. Their potential therapeutic benefits were also validated. However, larger placebo controlled blinded clinical trials will be required to establish the long term safety and efficacy profile of these therapies for MS. Their translation into the clinical practice can provide a new hope for the patients of this highly debilitating disease

Winery wastewater treatment for biomolecules recovery and water reuse purposes

Winery wastewater has a high pollutant load, but it also contains added-value molecules, such as phenolic compounds, which can be recovered. The recovery of these biomolecules has aroused great interest, providing benefits under different aspects, such as valorization of byproducts recovered from wastewater; reduction of the pollutant load of winery wastewater, facilitating its treatment and reuse in agriculture; reducing the environmental impacts of wineries; and promotion of the circular economy, due to the recovery of biomolecules and water reuse. Based on these matters, the chapter focuses on bringing relevant aspects about the generation of wastewater throughout the winemaking process, indicating the main biomolecules of commercial interest present in the wastewater, and proposing processes for the recovery of these biomolecules. In addition, the main technologies that are used in the treatment of winery wastewater are reported and discussed, envisaging the reuse of treated wastewater in agriculture.info:eu-repo/semantics/publishedVersio

Técnicas de Análise Espacial Aplicadas à Caracterização da População Idosa para Planear Cuidados de Enfermagem

O estudo aponta uma metodologia para caracterizar factores geodemográficos condicionantes da acessibilidade geográfica a cuidados de enfermagem, numa das regiões mais envelhecidas de Portugal, a partir do uso de Sistemas de Informação Geográfica. Analisam-se dados provenientes dos censos de população relativos aos idosos. Métodos: Utilizam-se técnicas de estatística espacial, para além das técnicas de estatística clássica. Estuda-se a autocorrelação espacial dos valores de alguns índices demográficos, caracterizando estatisticamente a existência de aglomerados na zona de estudo, recorrendo ao Índice Global de Moran. Igualmente caracteriza-se a concentração de valores elevados e de valores baixos para cada um dos índices, recorrendo ao cálculo da estatística GI*. Resultados: Identificam-se as regiões de maior envelhecimento, aquelas onde se perspectiva a maior utilização de recursos de Enfermagem por parte dos idosos. Conclusões: O estudo faz apelo à necessária gestão programática dos recursos materiais e humanos de enfermagem, perspectivando a minimização dos efeitos que a situação geodemográfica heterogénea condiciona

Students´ Representations of education Quality in Different School Levels: an Exploratory Study with Portuguese Students

Quality may be studied from different perspectives. Research shows that many variables may be envolved when the target is the educational system. One may analyse the motivation of students and teachers, the methodologies of teaching, the processes of evaluation, the resources available, the educational facilities, the course organization or the academic success. Recent research results show a strong positive correlation between students’ academic success and the quality of education available to them. However, the way students perceive the quality indicators has been a less explored area. We present the results of an academic longitudinal research project that aims to study the students’ representations of quality along the school system in order to contribute to an improvement of teaching and learning processes. The sampling of this exploratory study focused on compulsory education, secondary education and higher education. In this paper the authors analyse data collected in three institutions of higher education in Portugal, through a questionnaire survey