Study of antioxidant, antiproliferative and apoptosis-inducing properties of wild mushrooms from the Northeast of Portugal

Mushrooms are known as a powerful source of bioactive compounds including antioxidants, inhibitors of human tumour cell lines growth, inducers of apoptosis and enhancers of immunity. Indeed, many pre-clinical studies have been conducted in human tumour cell lines and in some cases a number of compounds isolated from mushrooms have followed to clinical trials. The Northeast of Portugal is one of the European regions with higher wild mushrooms diversity. However, to our knowledge, no studies had been conducted so far to verify their bioactivities. The main aim of this work was the evaluation of the bioactive properties (antioxidant properties and growth inhibitory potential on human tumour cell lines) of wild edible mushrooms collected in the Northeast of Portugal. Once properly identified, methanolic, ethanolic and boiling water extracts were prepared from thirty eight wild mushroom species collected in that region. Chemical characterization was obtained by high performance liquid chromatography (HPLC) coupled to a photodiode array detector (DAD) or to a refraction index detector (RI). Antioxidant activity assays were carried out in those extracts, including evaluation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging capacity, reducing power and inhibition of β-carotene bleaching. Extract-induced cell growth inhibition was assessed with the sulforhodamine B assay in four human tumour cell lines (NCI-H460 - lung cancer, MCF-7 -breast cancer, HCT-15 -colon cancer and AGS - gastric cancer). The effects on cell cycle profile and apoptosis were evaluated by flow cytometry and the effect on the expression levels of proteins related to cell cycle and apoptosis was further investigated by Western blotting. Three wild edible mushroom species revealed growth inhibitory activity in the studied human tumour cell lines: Clitocybe alexandri ethanolic extract, Lepista inversa methanolic extract and Suillus collinitus methanolic extract. C. alexandri ethanolic extract induced an S-phase cell cycle arrest and increased the percentage of apoptotic cells, in the NCI-H460 cell line. The analysed mushroom species also provided interesting antioxidant potential, mainly the boiling water extract of L. inversa which showed the highest DPPH radical scavenging activity, reducing power and β-carotene bleaching inhibition. S. collinitus methanolic extract induced a slight increase in the number of cells in G1, with a concomitant decrease in the percentage of cells in the S phase of the cell cycle and an increase in the percentage of apoptotic cells, in the MCF-7 cell line. The combined use of the S. collinitus methanolic extract and etoposide caused a greater decrease in the percentage of cell growth, when compared to either of them used individually, indicating the potential benefit of this combination. The tested extracts were chemically characterized and protocatechuic, p-hydroxybenzoic, p-coumaric and cinnamic acids were the main compounds identified on the phenolic (methanolic and ethanolic) extracts, while mannitol, trehalose and arabinose were the main sugars found in the polysaccharidic (boiling water) extracts after hydrolysis. The individual compounds identified in the extracts were submitted to a screening of tumour cells growth inhibitory activity, but only the phenolic acids and a related compound, cinnamic acid, presented activity. This compound was found to be the most potent one regarding cell growth inhibition in the NCI-H460 cell line. The effect of the individual and combined treatment with the identified compounds was also evaluated. Cinnamic and protochatequic acids caused a statistically significantly reduction in the number of viable cells. In addition, p-hydroxybenzoic acid did not show any significantly reduction in the viable cell number. Nevertheless, it was verified that the concomitant use of the three compounds provided the strongest decrease in the viable cell number, suggesting a possible concomitant effect of those compounds. Overall, the present work has contributed to further understand the bioactive potential of wild edible mushrooms from the Northeast of Portugal. This study allowed to identify some species with antioxidant or tumour cell growth inhibitory potential.

Information brokers and the use of budgetary and financial information by politicians: the case of Portugal

This paper analyses the intermediary role of the technical bodies that support the use of budgetary and financial information by central government politicians in Portugal. The main findings show that information brokers are playing a central role in preparing this information in a credible, simple and understandable way. However, even if not intentionally, the information they present can be biased. Politicians need to be aware that the information brokers they rely on may not be giving them ‘neutral’ information.

A conquista de Arzila pelos Portugueses : 1471

Em 1471 D. Afonso V (1432-1481), rei de Portugal, decidiu voltar ao Norte de África para dar continuidade à política de expansão africana da Coroa. O alvo escolhido foi Arzila e, em Agosto de 1471, a vila foi conquistada por uma hoste portuguesa que contou com 23.000 homens de combate. Pouco estudada pela historiografia, a conquista de Arzila é meritória de um lugar de destaque fruto da sua singularidade. Foi uma campanha meticulosamente planeada que incluiu a compra de toneladas de mantimentos e armas e o recrutamento da maior hoste até então levantada em Portugal. As consequências da queda de Arzila perduraram no tempo: Tânger foi abandonada dias depois e o tratado de tréguas assinado com o sultanato de Fez consolidou presença portuguesa na região, tendo esta permanecido imperturbável até ao início do século XVI, e permitiu a D. Afonso V abandonar de vez os campos de batalha norte-africanos e dar azo às suas ambições castelhanas. O presente estudo insere-se numa lacuna historiográfica face à pouca atenção que a conquista de Arzila granjeou, até ao momento, por parte dos historiadores portugueses. Assim sendo, é estudada não apenas a conquista de Arzila, enquanto acontecimento limitado ao mês de Agosto de 1471, mas também todos os preparativos que foram levados a cabo para permitirem o sucesso da campanha, bem como as consequências da queda de Arzila, mais prolongadas no tempo. Esse estudo é escorado não só nos relatos cronísticos mas também noutras fontes de tipo narrativo e na documentação de chancelaria. É através do cruzamento das várias fontes que é possível detectar e corrigir determinadas lacunas dos cronistas e dos historiadores que os seguiram.

An analysis of the effects of preferential trade policies through the estimation of quantitative models: the case of Portugal.

Doutoramento em Economia.

Satisfaction of users of primary health care with nursing care : sample of the central region of Portugal

Introdução: A satisfação dos doentes constitui um indicador indispensável para a avaliação da qualidade dos cuidados e há evidência da sua correlação com os resultados em saúde. A satisfação com os cuidados de saúde é um conceito multidimensional que considera aspetos como acesso, organização e interação doente - profissional. Consideramos que os cuidados de enfermagem, em particular, são fundamentais no processo saúde/doença. Objetivos: Validar uma escala para avaliar a satisfação dos utentes face aos cuidados de enfermagem, adaptado do instrumento EUROPEP e avaliar a satisfação dos utentes dos cuidados de saúde primários da região centro de Portugal. Material e métodos: Estudo transversal, com uma amostra de 827 utentes adultos (maioria do sexo feminino 64,4%) com uma média de idade de 50,08±18,58 anos. Os dados foram recolhidos através de um questionário, constituído por variáveis sociodemográficas, o instrumento EUROPEP (Ferreira, 1995) para avaliar a satisfação com os cuidados de saúde primários e para avaliar a satisfação especificamente com a equipa de enfermagem elaboramos questões adaptadas do instrumento EUROPEP e agrupadas nas dimensões relação de ajuda, dimensão interpessoal e instrumental. A consistência interna, reprodutibilidade e análise de conteúdo foram avaliados com recurso ao SPSS 23.0; considerando a consistência aceitável para um de Cronbach > 0,70. O coeficiente para cada item é apresentado com um intervalo de confiança de 95%. Resultados: Em todas as dimensões do questionário EUROPEP, a maior percentagem de satisfação com os cuidados situou-se entre “boa” e “muito boa”. As dimensões criadas para avaliar especificamente os cuidados de enfermagem apresentaram um coeficiente de α de Cronbach total de 0,972. Conclusões: Estes resultados sugerem que as dimensões criadas para avaliar os cuidados de enfermagem serão úteis para a investigação na população Portuguesa. A satisfação do utente é decisiva para a qualidade e eficiência dos cuidados prestados, sendo necessário o compromisso de todos os prestadores na implementação de práticas sistemáticas de gestão que conduzam à satisfação, dando particular atenção à melhoria contínua dos processos organizacionais. Palavras-chave: Satisfação dos Utentes; cuidados de saúde primários, cuidados de enfermagem, adulto, Portugal

Age and growth, mortality and reproduction of the striped sea bream, Lithognathus mormyrus Linnaeus 1758, from the south coast of Portugal (Algarve)

The striped sea bream, Lithognathus mormyrus, used for this population dynamics study were obtained from longline catches and market sampling in the Algarve (south Portugal). The macroscopic analysis of the gonads and the gonad somatic index showed that the south Portuguese population of L. mormyrus spawns mainly between late spring and summer (June to August). The length at first maturity was similar for males and females and the value for both sexes combined was estimated to be 16.08 cm, corresponding to an age between 1 and 2 years. Fish age classes (0 to 13) were determined by reading growth rings on whole otoliths. Age determination was validated by marginal increment analysis. The estimated parameters of the von Bertalanffy growth equation were L infinity = 35.30 cm, K = 0.264 and t(0) = -0.809. Mortality rates were calculated for fish captured with longlines, and the estimated parameters were M = 0.356, Z = 0.622 and F = 0.266. From an Algarve fishery management perspective, these results suggest the need for an increase in the minimum landing size (from 15 to 17 cm), which should be beneficial for the sustainability and conservation of this species. The results also showed that fishing with longlines off the Algarve coast may allow for a sustainable use of the resource.

Age and growth, mortality, reproduction and relative yield per recruit of the bogue, Boops boops Linne, 1758 (Sparidae), from the Algarve (south of Portugal) longline fishery

Samples of Boops boops ranging from 7.4 to 30.5 cm were obtained mainly by longline, supplemented by beach seining in the Ria Formosa lagoon, and by market sampling in the Algarve (southern Portugal). The macroscopic analyses of the gonads and the gonad somatic index showed that the south coast of Portugal B. boops spawn mainly from late winter to spring, between February and May. The length at first maturity was similar for males and females and the value for both sexes combined was estimated to be 15.22 cm, corresponding to an age range of 1-3. Age was determined by reading growth bands on otoliths. Age determination was validated by marginal increment analysis. The estimated parameters were L-infinity = 28.06, K = 0.22 and t(0) = -1.42. Mortality rates were calculated for fish captured with longlines, and the estimated parameters were M = 0.33, Z = 1.04 and F = 0.71. Relative yield per recruit analysis and sensitivity analysis showed that the resource is moderately exploited. From the perspective of sustainability, these results provide support for the use of longlines as a gear that is among the least harmful for species such as the bogue.

An assessment of catches and harvest of recreational shore angling in the north of Portugal

Recreational shore angling in the Atlantic Ocean between Moledo and Aveiro (Portugal) was examined using roving creel surveys (March/September 2001). Cooperation was high (90% of 2310 anglers approached). At least 39 species of fish were caught at a rate of approximately 0.5 fish angler h(-1). An estimated 7319 kg of Dicentrarchus labrax (L.), with 45.6% below the minimum legal size, and 2040 kg of sea breams (genus Diplodus), correspond to 5.75% and 1.19% of the commercial landings in the same geographical area, respectively. The results shed light on a number of issues relevant to integrated coastal management, including temporal and spatial distribution of fishing effort, species caught, sizes of fish, catch rates, and factors influencing catches and angler satisfaction. Whilst the results suggest that the catches of sea breams and sea bass by day-time recreational shore angling in northern Portugal are small compared with commercial fishing, other recreational activities, such as boat fishing and spear-fishing, must be assessed.

Diet of the blue marlin, Makaira nigricans, off the south coast of Portugal

The blue marlin, Makaira nigricans (Istiophoridae), is a large, top predator with a worldwide distribution whose feeding ecology is still unknown in the northeast Atlantic. The stomach contents of 24 Atlantic blue marlin, caught by the Big Game fishing fleet (between 2007 and 2010) off the south coast of Portugal, were studied. All marlin fed exclusively on pelagic fish, with a total of 180 prey items recorded, belonging to 6 identified species. The most important family in the blue marlin diet was Scombridae, representing 70% of the total prey items. Among the Scombridae, the chub mackerel, Scomber colias, was the most important prey species (47.9% by weight, 51.1% by number, 58.3% by frequency of occurrence). The length of prey found in the stomachs ranged from 136 to 727 mm in length, and a significant positive correlation was found between prey and blue marlin size. Our results suggest that in this geographical area (NE Atlantic) marlin feed on a small spectrum of species, with a preference for foraging on seamounts.

Feeding habits of the two-banded sea bream (Diplodus vulgaris) and the black sea bream (Spondyliosoma gantharus) (Sparidae) from the south-west coast of Portugal

The stomach contents of Diplodus vulgaris and Spondyliosoma cantharus were: analysed using three simple methods (numeric, gravimetric and frequency of occurrence) and a composite index (I.R.I - Index of Relative Importance). To compare the species, the Schoener index was used. The diet of D. vulgaris consisted mainly of ophiuroids, polychaetes, amphipods and echinoids, while polychaetes, amphipods and hydrozoans dominated in the case of S. cantharus. There were some size-related differences in S. cantharus feeding. Diet overlap was relatively slight, with significant differences in feeding between the two species, notably in terms of greater consumption of echinoderms by D. vulgaris and hydrozoans by S. cantharus. As is the case for the majority of sea breams, D. vulgaris and S. cantharus are characterised by a diverse diet in terms of prey reflecting available prey items in their environment.

Reproductive biology of the blackmouth catshark, Galeus melastomus (Chondrichthyes : Scyliorhinidae) off the south coast of Portugal

The reproductive biology of 1045 female (9.5-67 cm (total length(TL)) and 1007 male (9.4-64.3 cm TL) blackmouth catsharks, Galeus melastomus, was investigated. The sharks were caught off southern Portugal by bottom crustacean trawlers at depths from 209 to 754 m. The sex ratio was 1:1, and this species is sexually dimorphic with males approaching maturity at smaller size than females. Sexual segregation appears to be given for the stork within the study area. Sexual maturity was reached at a total length above 49 cm in males and above 56 cm in females. Mating and egg-deposition take place all year round, with two reproductive peaks of activity, in winter and summer. Egg capsules are, on average, 54 mm long and 21 mm wide, with a maximum of 63 x 25 mm encountered. Morphometric measurements of claspers, testes, ovaries, and oviducal glands were suitable for determining sexual maturity of blackmouth catshark.

The reproductive biology of Spondyliosoma cantharus (L.) from the SW Coast of Portugal

The study of Spondyliosoma cantharus (L.) reproduction was carried out within the framework of a project on the Fisheries resources of the south-west coast of Portugal, and was based on the analysis of the spawning season, gonad maturation, size-at-maturity, fecundity, and hermaphroditism. Spawning took place from February to April, peaking in March. Analysis of the sex ratio by size class and season showed that females were more abundant throughout the year (M/F=0.57) and in the smaller size classes. Overall size at first maturity (L-50) was 20.10 cm total length (TL), with a significant difference between males (22.41 cm, TL) and females (19.98 cm, TL). Absolute fecundity (Fa) ranged from 37,506 to 112,074 oocytes, with a mean of 61,396. A power type relationship best described the relationships between absolute fecundity and TL (Fa = 436.27TL(1.575)), and somatic weight (Fa = 2979.7SW(0.585)). The number of oocytes/g of female somatic weight ranged from 217 to 549, with a mean of 346. The reproductive strategy of this species is characterised by protogynic hermaphroditism, as indicated by the presence of individuals in transition and of testes with vestiges of preceding ovaries associated with the significant differences in the size frequency distributions of the sexes.

The reproductive biology of the two-banded sea bream (Diplodus vulgaris) from the southwest coast of Portugal

The study of the reproduction of Diplodus vulgaris (Geoff.) as part of a base-line study of the fishery resources of the southwest coast of Portugal, was based on the analysis of the spawning season, gonad maturation, size at maturity, fecundity and hermaphroditism. The spawning season is relatively long, from December to March, with peaks in January and February. No significant differences were found either in the sex ratio (M:F = 1.01) over the year or by size. The size at first maturity (L-50) for all sexes and undetermined individuals combined was 18.33 cm total length (TL), with no significant differences between males and females. The estimated L-50 is considerably greater than the minimum legal size in Portugal of 15.0 cm. Mean absolute fecundity (F-a) was 131 127 oocytes, ranging from 31 523 to 250 608. The relationship between absolute fecundity and total length (TL) (F-a = 25 398 TL-484 426) and somatic weight (SW) (F-a = 878.8SW-71 416) was of the linear type. The mean number of oocytes per gram of somatic weight was 526, ranging from 194 to 887. The reproductive strategy of this species is characterized by a rudimentary hermaphroditism with possible protandry, as evidenced by the existence of individuals in sexual transition.