Produção de voláteis por culturas de raízes transgénicas de Anethum graveolens:influência da regeneração e da capacidade de biotransformação

Tese de mestrado, Biologia (Biologia Celular e Biotecnologia), 2007, Universidade de Lisboa, Faculdade de CiênciasForam usadas culturas de raízes transgénicas (r.t.) de Anthum graveolens para testar a possibilidade de regeneração e para avaliar a influência da capacidade de biotransformação dos álcoois monoterpénicos, mentol e geraniol, no crescimento e produção de voláteis. A regeneração foi ensaiada em fotoperíodo, de 24h luz em jiffy pellets e de 16h luz em caixas de Petri com 20 ml de meio MS/2. Parte destas foi suplementada com higromicina B (2 mg.l-1, 5 mg.l-1 e 7 mg.l-1) ou ácido salicílico (0,3; 0,5 e 0,8 mM). Outras foram tratadas com radiação UV-C (30 min, 30+15 min, 30+2x15 min, 30+3x15 min), posteriormente à inoculação com extremidades de r.t. A influência da biotransformação foi analisada pela adição de substrato às culturas 15 dias após a inoculação, numa concentração de 25 mg.l-1. As culturas foram mantidas em meio SH, sob escuridão permanente a 24ºC, com agitação a 80 r.p.m. A avaliação do crescimento foi efectuada pelo método da desassimilação e pela medição do peso fresco e peso seco das raízes ao longo de 7 semanas. A componente volátil, extraída por destilaçãoextracção, foi analisada por CGL e CGL-EM. A regeneração não foi conseguida. As r.t. submetidas a fotoperíodo de 16h luz revelaram crescimento, reverdecimento e formação de tecido caloso. Estas respostas não foram observadas com a aplicação de higromicina B ou ácido salicílico nem com o ensaio em jiffy pellets. Estas condições parecem ser prejudiciais ao crescimento. A componente volátil constitutiva das r.t. de A. graveolens é composta, em mais de 50%, por falcarinol (9- 52%), apiole (9-24%), ácido palmítico (7-16%), ácido linoleico (4-9%), miristicina (2-8%) e n-octanal (2-8%). A adição de mentol ou geraniol não influenciou substancialmente o crescimento. Após a adição, a proporção relativa de mentol decresceu rapidamente num período de 48h e observou-se a concomitante produção de acetato de mentilo. O geraniol adicionado foi biotransformado em, pelo menos, 10 produtos, os álcoois linalol, α-terpineol e citronelol, os aldeídos neral e geranial, os ésteres acetato de citronelilo, de nerilo e de geranilo e os óxidos de nerol e linalol. A proporção relativa de geraniol decresceu rapidamente num período de 48h em detrimento dos produtos. A concentração relativa de todos os produtos de biotransformação detectados diminuiu gradualmente ao longo do tempo de culturaAnethum graveolens hairy root cultures were used to assess regeneration and evaluate the influence of the biotransformation capacity of the monoterpene alcohols, menthol and geraniol, on growth and production of volatile compounds. Regeneration was assayed in photoperiod, 24h light in jiffy pellets and 16h light in Petri dishes with 20 ml of MS/2 medium. Some of these were supplemented with higromycin B (2 mg.l-1, 5 mg.l-1 e 7 mg.l-1) or salicylic acid (0,3; 0,5 e 0,8 mM). Others were subjected to UV-C radiation (30 min, 30+15 min, 30+2x15 min, 30+3x15 min) after inoculation with hairy root tips. The biotransformation capacity was assayed by adding substrate, 25 mg.l-1, to the cultured roots 15 days after inoculation. The cultures were maintained in SH medium, in permanent darkness, at 24ºC, and 80 r.p.m. Hairy root growth was measured by the dissimilation method and by fresh and dry weight measurement, throughout 7 weeks. The volatile component, extracted by distillation-extraction, was analyzed by GC and GC-MS. Regeneration was not achieved. The 16h light photoperiod roots showed growth, regreening and calli formation. These responses were not observed in higromycin B or salicylic acid treated roots or in jiffy pellets. These conditions appear to be disadvantageous (harmful) to growth. The constitutive volatile component of A. graveolens hairy roots is composed, in more than 50%, by falcarinol (9-52%), apiole (9-24%), palmitic acid (7-16%), linoleic acid (4-9%), miristicin (2-8 %) and n-octanal (2-8%). The addition of menthol or geraniol did not substantially influence growth. After addition, the relative concentration of menthol quickly decreased in a period of 48h, and a concomitant production of menthyl acetate was observed. This was the only detectable biotransformation product and its relative amount diminished gradually throughout culture time. The added geraniol was biotransformed in, at least, 10 products, the alcohols linalool, α-terpineol and citronellol, the aldehydes neral and geranial, the esters citronellyl, neryl and geranyl acetates and linalool and nerol oxides. The relative proportion of geraniol quickly decreased in a period of 48h at the expense of the products. The relative concentration of all the detected biotransformation products decreased gradually over time in cultur

Integrating factors associated with complex wound healing into a mobile application: findings from a cohort study

Complex, chronic or hard-to-heal wounds are a prevalent health problem worldwide, with significant physical, psychological and social consequences. This study aims to identify factors associated with the healing process of these wounds and develop a mobile application for wound care that incorporates these factors. A prospective multicentre cohort study was conducted in nine health units in Portugal, involving data collection through a mobile application by nurses from April to October 2022. The study followed 46 patients with 57 wounds for up to 5 weeks, conducting six evaluations. Healing time was the main outcome measure, analysed using the Mann–Whitney test and three Cox regression models to calculate risk ratios. The study sample comprised various wound types, with pressure ulcers being the most common (61.4%), followed by venous leg ulcers (17.5%) and diabetic foot ulcers (8.8%). Factors that were found to impair the wound healing process included chronic kidney disease (U = 13.50; p = 0.046), obesity (U = 18.0; p = 0.021), non-adherence to treatment (U = 1.0; p = 0.029) and interference of the wound with daily routines (U = 11.0; p = 0.028). Risk factors for delayed healing over time were identified as bone involvement (RR 3.91; p < 0.001), presence of odour (RR 3.36; p = 0.007), presence of neuropathy (RR 2.49; p = 0.002), use of anti-inflammatory drugs (RR 2.45; p = 0.011), stalled wound (RR 2.26; p = 0.022), greater width (RR 2.03; p = 0.002), greater depth (RR 1.72; p = 0.036) and a high score on the healing scale (RR 1.21; p = 0.001). Integrating the identified risk factors for delayed healing into the assessment of patients and incorporating them into a mobile application can enhance decision-making in wound care.info:eu-repo/semantics/publishedVersio

Diabetes and Cardiovascular Disease: prognosis five years after percutaneous revascularization

Fundamentos: O diabetes mellitus (DM) é reconhecidamente fator de risco cardiovascular. Sabendose que a intervenção coronariana percutânea (ICP) melhora o prognóstico da doença coronariana (DC), pretendemos verificar se esse efeito é similar em doentes diabéticos (D) e não diabéticos (ND). Objetivo: Analisar o prognóstico em longo prazo do DM em pacientes submetidos a ICP. Métodos: Estudo de coorte, unicêntrico, retrospectivo, envolvendo pacientes consecutivos submetidos à ICP, eletiva ou de urgência, entre janeiro 2002 e dezembro 2003. Definiram-se dois grupos: pacientes com DM (D) e sem DM (ND). Compararam-se as variáveis clínicas e angiográficas da ICP com o resultado clínico ao final de cinco anos. Definiram-se como eventos maiores cardiovasculares (EMC): morte, nova síndrome coronariana aguda, acidente vascular encefálico (AVE) e nova revascularização cirúrgica ou ICP. Foram ainda avaliadas as taxas de trombose de stent, revascularização do vaso-alvo (RVA) e revascularização da lesão-alvo (RLA). Resultados: O seguimento em cinco anos foi 94%. Foram realizadas 446 ICP em 406 pacientes, média de idade =63,0±11 anos, 70,4% masculino. Destes, 128 (31,5%) eram do grupo D. Em cinco anos o valor de EMC foi 50,7% para D e 36,7% para N. Encontrou-se mortalidade global de 28,1% vs. 14,4% (p=0,0016) e mortalidade cardiovascular de 15,6% vs. 8,3% (p=0,039), respectivamente, para D vs. ND. Os restantes parâmetros não foram estatisticamente significativos. Não houve diferenças em termos de trombose de stent e tipos de stent utilizados nos dois grupos.Background: Diabetes Mellitus (DM) is a recognized cardiovascular risk factor, and knowing that percutaneous coronary intervention (PCI) improves prognoses of coronary artery disease (CAD), we intend to ascertain whether this effect is similar in diabetic (D) and nondiabetic (ND) patients. Objective: To analyze the long term impact of DM in patients undergoing PCI. Methods: Retrospective unicentric cohort study of consecutive patients undergoing elective or urgent PCI between January 2002 and December 2003, divided into two groups: diabetic (D) and non-diabetic (ND). Clinical, angiographic and coronary intervention PCI profiles were compared to clinical outcome at 5 years. Major cardiovascular events (MACE) were defined as death, new acute coronary syndrome (ACS), stroke (S), and new surgical (CABG) or percutaneous (PCI) revascularization. The stent thrombosis, target vessel revascularization (TVR) and target lesion revascularization (TLR) rates were also evaluated. Results: The five year follow-up rate was 94%. During the study, 446 PCIs were performed on 406 patients; 63.0±11 years old, 70.4% male. Among them, 128 (31.5%) were D. At the five-year follow-up, the MACE rate was 50.7% for D and 36.7% for ND (p=0.01). The global mortality rates were 28.1% vs 14.4% (p=0.0016) and the cardiovascular mortality rates were 15.6% vs. 8.3% (p=0.039) for D and ND respectively. The remaining parameters were not statistically significant. No differences were noted between stent thrombosis and the stent types used in both groups.info:eu-repo/semantics/publishedVersio

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio

MAMMALS IN PORTUGAL : A data set of terrestrial, volant, and marine mammal occurrences in P ortugal

Mammals are threatened worldwide, with 26% of all species being includedin the IUCN threatened categories. This overall pattern is primarily associatedwith habitat loss or degradation, and human persecution for terrestrial mam-mals, and pollution, open net fishing, climate change, and prey depletion formarine mammals. Mammals play a key role in maintaining ecosystems func-tionality and resilience, and therefore information on their distribution is cru-cial to delineate and support conservation actions. MAMMALS INPORTUGAL is a publicly available data set compiling unpublishedgeoreferenced occurrence records of 92 terrestrial, volant, and marine mam-mals in mainland Portugal and archipelagos of the Azores and Madeira thatincludes 105,026 data entries between 1873 and 2021 (72% of the data occur-ring in 2000 and 2021). The methods used to collect the data were: live obser-vations/captures (43%), sign surveys (35%), camera trapping (16%),bioacoustics surveys (4%) and radiotracking, and inquiries that represent lessthan 1% of the records. The data set includes 13 types of records: (1) burrowsjsoil moundsjtunnel, (2) capture, (3) colony, (4) dead animaljhairjskullsjjaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8),observation in shelters, (9) photo trappingjvideo, (10) predators dietjpelletsjpine cones/nuts, (11) scatjtrackjditch, (12) telemetry and (13) vocalizationjecholocation. The spatial uncertainty of most records ranges between 0 and100 m (76%). Rodentia (n=31,573) has the highest number of records followedby Chiroptera (n=18,857), Carnivora (n=18,594), Lagomorpha (n=17,496),Cetartiodactyla (n=11,568) and Eulipotyphla (n=7008). The data setincludes records of species classified by the IUCN as threatened(e.g.,Oryctolagus cuniculus[n=12,159],Monachus monachus[n=1,512],andLynx pardinus[n=197]). We believe that this data set may stimulate thepublication of other European countries data sets that would certainly contrib-ute to ecology and conservation-related research, and therefore assisting onthe development of more accurate and tailored conservation managementstrategies for each species. There are no copyright restrictions; please cite thisdata paper when the data are used in publications.info:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Mammals in Portugal: a data set of terrestrial, volant, and marine mammal occurrences in Portugal

Mammals are threatened worldwide, with ~26% of all species being included in the IUCN threatened categories. This overall pattern is primarily associated with habitat loss or degradation, and human persecution for terrestrial mammals, and pollution, open net fishing, climate change, and prey depletion for marine mammals. Mammals play a key role in maintaining ecosystems functionality and resilience, and therefore information on their distribution is crucial to delineate and support conservation actions. MAMMALS IN PORTUGAL is a publicly available data set compiling unpublished georeferenced occurrence records of 92 terrestrial, volant, and marine mammals in mainland Portugal and archipelagos of the Azores and Madeira that includes 105,026 data entries between 1873 and 2021 (72% of the data occurring in 2000 and 2021). The methods used to collect the data were: live observations/captures (43%), sign surveys (35%), camera trapping (16%), bioacoustics surveys (4%) and radiotracking, and inquiries that represent less than 1% of the records. The data set includes 13 types of records: (1) burrows | soil mounds | tunnel, (2) capture, (3) colony, (4) dead animal | hair | skulls | jaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8), observation in shelters, (9) photo trapping | video, (10) predators diet | pellets | pine cones/nuts, (11) scat | track | ditch, (12) telemetry and (13) vocalization | echolocation. The spatial uncertainty of most records ranges between 0 and 100 m (76%). Rodentia (n =31,573) has the highest number of records followed by Chiroptera (n = 18,857), Carnivora (n = 18,594), Lagomorpha (n = 17,496), Cetartiodactyla (n = 11,568) and Eulipotyphla (n = 7008). The data set includes records of species classified by the IUCN as threatened (e.g., Oryctolagus cuniculus [n = 12,159], Monachus monachus [n = 1,512], and Lynx pardinus [n = 197]). We believe that this data set may stimulate the publication of other European countries data sets that would certainly contribute to ecology and conservation-related research, and therefore assisting on the development of more accurate and tailored conservation management strategies for each species. There are no copyright restrictions; please cite this data paper when the data are used in publications

Approaches in biotechnological applications of natural polymers

Natural polymers, such as gums and mucilage, are biocompatible, cheap, easily available and non-toxic materials of native origin. These polymers are increasingly preferred over synthetic materials for industrial applications due to their intrinsic properties, as well as they are considered alternative sources of raw materials since they present characteristics of sustainability, biodegradability and biosafety. As definition, gums and mucilages are polysaccharides or complex carbohydrates consisting of one or more monosaccharides or their derivatives linked in bewildering variety of linkages and structures. Natural gums are considered polysaccharides naturally occurring in varieties of plant seeds and exudates, tree or shrub exudates, seaweed extracts, fungi, bacteria, and animal sources. Water-soluble gums, also known as hydrocolloids, are considered exudates and are pathological products; therefore, they do not form a part of cell wall. On the other hand, mucilages are part of cell and physiological products. It is important to highlight that gums represent the largest amounts of polymer materials derived from plants. Gums have enormously large and broad applications in both food and non-food industries, being commonly used as thickening, binding, emulsifying, suspending, stabilizing agents and matrices for drug release in pharmaceutical and cosmetic industries. In the food industry, their gelling properties and the ability to mold edible films and coatings are extensively studied. The use of gums depends on the intrinsic properties that they provide, often at costs below those of synthetic polymers. For upgrading the value of gums, they are being processed into various forms, including the most recent nanomaterials, for various biotechnological applications. Thus, the main natural polymers including galactomannans, cellulose, chitin, agar, carrageenan, alginate, cashew gum, pectin and starch, in addition to the current researches about them are reviewed in this article.. }To the Conselho Nacional de Desenvolvimento Cientfíico e Tecnológico (CNPq) for fellowships (LCBBC and MGCC) and the Coordenação de Aperfeiçoamento de Pessoal de Nvíel Superior (CAPES) (PBSA). This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and COMPETE 2020 (POCI-01-0145-FEDER-006684) (JAT)