Phylloseptin-PBa—A Novel Broad-Spectrum Antimicrobial Peptide from the Skin Secretion of the Peruvian Purple-Sided Leaf Frog (Phyllomedusa Baltea) Which Exhibits Cancer Cell Cytotoxicity

Antimicrobial peptides from amphibian skin secretion display remarkable broad-spectrum antimicrobial activity and are thus promising for the discovery of new antibiotics. In this study, we report a novel peptide belonging to the phylloseptin family of antimicrobial peptides, from the skin secretion of the purple-sided leaf frog, Phyllomedusa baltea, which was named Phylloseptin-PBa. Degenerate primers complementary to putative signal peptide sites of frog skin peptide precursor-encoding cDNAs were designed to interrogate a skin secretion-derived cDNA library from this frog. Subsequently, the peptide was isolated and identified using reverse phase HPLC and MS/MS fragmentation. The synthetic replicate was demonstrated to have activity against S. aureus, E. coli and C. albicans at concentrations of 8, 128 and 8 mg/L, respectively. In addition, it exhibited anti-proliferative activity against the human cancer cell lines, H460, PC3 and U251MG, but was less active against a normal human cell line (HMEC). Furthermore, a haemolysis assay was performed to assess mammalian cell cytotoxicity of Phylloseptin-PBa. This peptide contained a large proportion of α-helical domain, which may explain its antimicrobial and anticancer activities.

Atmospheric cold plasma process for vegetable leaf decontamination: A feasibility study on radicchio (red chicory, Cichorium intybus L.)

Cold plasma is an emerging non-thermal processing technology that could be used for large scale leaf decontamination as an alternative to chlorine washing. In this study the effect of an atmospheric cold plasma apparatus (air DBD, 15 kV) on the safety, antioxidant activity and quality of radicchio (red chicory, Cichorium intybus L.) was investigated after 15 and 30 min of treatment (in afterglow at 70 mm from the discharge, at 22 °C and 60% of RH) and during storage. Escherichia coli O157:H7 inoculated on radicchio leaves was significantly reduced after 15 min cold plasma treatment (-1.35 log MPN/cm²). However, a 30 min plasma treatment was necessary to achieve a significant reduction of Listeria monocytogenes counts (-2.2 log CFU/cm²). Immediately after cold plasma treatment, no significant effects emerged in terms of antioxidant activity assessed by the ABTS and ORAC assay and external appearance of the radicchio leaves. Significant changes between treated and untreated radicchio leaves are quality defects based on the cold plasma treatment. Atmospheric cold plasma appears to be a promising processing technology for the decontamination of leafy vegetables although some criticalities, that emerged during storage, need to be considered in future studies.

Salinity-induced effects on transpiration rate, stomatal conductance and leaf area of three olive (Olea europaea L.) varieties

Transpiration of two year-old olive trees of three different varieties, Arbequina, Cobrançosa and Galega (18 trees per variety), irrigated with three levels of salt (0, 80 or 200 mM NaCl) for about 90 days, was measured by a gravimetric method. To determine leaf area, each tree was photographed from the side against a white background and the total area of each projected image was determined with ImageJ software. To calibrate these area determinations, one tree of each variety was subsequently stripped of all its leaves and its total leaf area was accurately measured. A correlation was then obtained between the area on the photograph of this particular tree and the total area of the detached leaves of the same tree. Using the leaf area determined by this procedure, transpiration rates of the trees could be calculated. Knowing leaf and air temperatures and RH, it was possible to determine the difference in molar fraction of water between the leaf and the air. Using this and the values of the transpiration rate, stomatal conductance could be calculated (gs calc) and compared with the conductance measured on the same trees with a porometer (gs). Actual leaf area of a plant was 1,40 (Arbequina), 1,42 (Cobrançosa) or 1,24 (Galega) times the area measured with ImageJ on the photograph of the same plant. Leaf area of the trees, on average of all salt irrigations, was significantly higher on Arbequina (0,187 m2) then on the other two varieties (0,138 m2 or 0,148 m2, for Cobrançosa or Galega, respectively), but did not differ significantly in percentage of controls (0 salt). On average of all three varieties, leaf area was also higher on plants irrigated without salt (0,181 m2) than on plants exposed to 80 or 200 mM NaCl (0,152 m2 or 0,140 m2, respectively), which did not differ between them. The same significant difference was observed when leaf area was expressed as percentage of controls. Transpiration rate was significantly higher on Cobrançosa (1,17 mmol m-2 s-1), on average of all treatments, but there were no significant differences between Arbequina (1,08 mmol m-2 s-1) and Galega (0,82 mmol m-2 s-1). In percentage of controls, there were no significant differences between varieties. Salt reduced significantly the transpiration rate in all varieties, both the actual and percentual values, to about 50% or 30% of controls when exposed to 80 mM or 200 mM NaCl, respectively. Stomatal conductance (gs), assessed by porometry, was significantly higher in control plants, mainly in Cobrançosa (102 mmol m-2 s-1), then in Arbequina (77 mmol m-2 s-1) and the lower values were found in Galega (51 mmol m-2 s-1). Salt reduced gs, on average of the three varieties to 30% or 10% of controls on exposure to 80 mM or 200 mM NaCl, respectively. Calculated (gs calc) and measured (gs) values of stomatal conductance showed a close relation between them (0,967, R2 = 0,837) which indicates this non-destructive method to determine whole-plant leaf area to be reasonably accurate.

Assessment of higher plant species suitability for ecological restoration following major disturbance

O presente trabalho é constituído por uma revisão bibliográfica e uma série de estudos científicos experimentais visando a melhoria do conhecimento da ecologia de germinação e resposta ao estresse hídrico em arbustos autóctones. Estes estudos pretendem contribuir para o conhecimento da ecologia de espécies autóctones na fase de germinação e estabelecimento inicial das plântulas através da comparação da resposta de sementes de diferentes proveniências. Os primeiros três capítulos focam-se no estudo dos efeitos de pré-tratamentos na germinação de espécies arbustivas com diferentes estratégias de regeneração e dormência da semente, enquanto os dois últimos capítulos incidem sobre o efeito de diferentes condições de disponibilidade de água na germinação e estabelecimento inicial de plântulas com potencial para restauro ecológico. Este estudo procura abordar aspetos importantes para o estudo e validação do uso de espécies alvo para o restauro ecológico em zonas Mediterrânicas, contribuindo deste modo para a melhoria do conhecimento da ecologia destas espécies. A germinação foi estimulada por pré-tratamentos de calor em quatro das cinco espécies cuja época de dispersão da semente na primavera-verão. As sementes provenientes do sul tenderam a germinar melhor, sendo ao mesmo tempo mais pequenas. Por outro lado, a germinação em espécies cuja dispersão da semente ocorre no outono-inverno variou de acordo com o tipo de dormência da semente. A germinação foi em geral favorecida pela estratificação a frio na espécie apresentando dormência fisiológica mas foi negativamente afetada na espécie que não apresenta dormência. Esta resposta não foi, no entanto, inequívoca pois foi dependente da proveniência estudada. O efeito da estratificação a frio teve em geral um efeito mais positivo ou menos negativo nas sementes provenientes do norte em comparação com as outras proveniências. O stress-osmótico teve um efeito negativo consistente na germinação de Pistacia lentiscus e outro fator que também afetou significativamente a germinação foi a variabilidade intra-populacional entre plantas mãe. As duas proveniências apresentaram diferentes graus de associação entre a germinação e/ou caracteres morfológicos da semente ou planta mãe. Quanto à fase de estabelecimento inicial, o baixo conteúdo em água teve um claro efeito negativo no investimento relativo em biomassa acima do solo e um efeito positivo no investimento relativo em biomassa abaixo do solo em Arbutus unedo. Esta resposta não foi inequívoca, uma vez que as proveniências diferiram em adaptações morfológicas ao baixo conteúdo em água. As plântulas da proveniência mais húmida revelaram uma taxa de crescimento relativo superior mas, ao mesmo tempo, uma fraca adaptação às condições de baixa disponibilidade de água, quando comparadas com as outras proveniências. Ao contrário, as plântulas da proveniência seca com verão mais quente apresentaram semelhantes diâmetro do colo da raiz, peso seco de folhas e performance fisiológica sob os dois regimes de irrigação. Os resultados obtidos revelam diferenças significativas na germinação e desenvolvimento inicial entre as distintas proveniências que poderão estar relacionadas com adaptações ao clima dos locais de origem. Os desenvolvimentos futuros do estudo dos mecanismos subjacentes às adaptações observadas poderiam contribuir para a melhoria das previsões de sucesso do estabelecimento inicial em diferentes populações de plantas, permitindo aumentar a confiança e efetividade de custos nas decisões relacionadas com ações de restauro ecológico em cenários de alterações climáticas.

Ecological quality assessment in transitional systems

Estuaries are poles of attraction for human settlement which is a source of pressures to surface water bodies. The implementation of the European Water Framework Directive (WDF, 2000/60/EC) has increased the investigation in order to develop methodologies to assess the Ecological Quality Status (EQS) of aquatic ecosystems. Transitional systems are naturally stressed and characterized by highly dynamic physical, chemical and hydro-morphologic conditions and by species with a higher level of tolerance to change, being more difficult to develop suitable quality indicators for these systems. The general purpose of this study is to test the ability of synthesis descriptors, including primary (S, taxa richness) and derived biological variable (H’, Shannon-Wiener diversity), biotic indices (AMBI and M-AMBI), body size properties (abundance distribution by body size classes, length, weight and length-weight relationships) and non-taxonomic indices (ISS), as well as functional indicators related to the decomposition rates of various experimental substrates, a macrophyte (Phragmites australis) and an alga (Fucus vesiculosus), to evaluate the environmental quality in transitional systems. This study was carried out in one of the most pristine channels of the Ria the Aveiro, Mira Channel, along a full salinity gradient and in a metals and metalloid sediment contamination area, the Estarreja Channel, and two reference channels (Canelas and Salreu). In this study were used different sampling techniques, the leaf-bag technique and a hand-held corer. In Mira Channel, the alga and the macrophyte presented an opposite trend in the decomposition rate along the salinity gradient, with the decomposition rates of the alga always higher than those of the macrophyte. The decomposition rates of the macrophyte and the alga were higher in the mid estuary and in higher salinity areas, respectively, corresponding to the preferencial distribution areas of each species. The macrobenthic fauna associated with the decaying and an artificial substrate (control) showed equally well the benthic succession from the marine to the freshwater areas and, despite the strong differences in the decay rates, no significant differences were found between the benthic communities associated with the alga and the macrophyte. The body size properties of the macrobenthic fauna associated with the P. australis leaf-bag (1mm and 5mm) and corer samples were studied along the full salinity gradient. The dominant species of the sub-set of measured specimens were not the same of the original macrobenthic fauna sampled but, despite that, the sub-set of measured specimens was also able to show the benthic succession from the marine to the freshwater areas. The body size abundance distribution of the benthic macroinvertebrates according to the ISS size classes did not show a particular trend in any sampler along the salinity gradient. Significant differences were found in the length, weight and length-weight relationships of Annelids, , Molluscs and even some species along the salinity gradient. No significant differences were found in the AMBI, M-AMBI and ISS values along the salinity gradient for all the samplers. The EQS of the corer samples obtained using the M-AMBI was lower than that of the leaf-bags. The EQS obtained with the ISS was higher than that obtained with the M-AMBI in the leaf-bags but not in the corer samples. The ecological effects of contaminated sediments associated with the industrial chemical effluents discharged in the Estarreja Channel were studied a decade after ceasing the emissions, using the Sediment Quality Triad approach and two reference channels. The results showed that despite the emissions ceased in 2004, the sediment remains polluted with high levels of metals and metalloid, available to bioaccumulation and with severe consequences at the community level. The sediment contamination problem was also studied using the leaf-bag technique with a macrophyte, an alga and a control substrate. The results showed that the decay rates, the associated macrofauna and the application of the AMBI, M-AMBI and ISS indices to the mesh-bag samples were not able to identify the sediment contamination. Contrarily to the AMBI, the M-AMBI and the ISS showed significant differences between the contaminated and the reference channels for the corer samples. Although such statistical significance, the interest of using these complex biotic indices could be questioned, when much simple ones, like the S and H’ allow to reach the same conclusions.

Kids just wanna have fun: Children's experiences of a weight management programme

Objectives: To explore children's accounts of their experiences of the UK‘s largest childhood obesity programme, MEND (Mind, Exercise, Nutrition…Do it!) (See www.mendprogramme.org). Design: Semi-structured interviews were conducted with children who had completed the MEND obesity programme. Interviews were transcribed verbatim and analysed using Interpretative Phenomenological Analysis (IPA). Method Fourteen children spanning diverse areas of London comprised this study (eight male, six female), aged between 11 and 14 years and in secondary school. Participants were interviewed a year after completing one of the London-based MEND obesity programmes. Results: This article focuses on the most common and striking theme to emerge from the original dataset (The complete analysis may be found in L. Watson, Unpublished doctoral thesis): Fun. Subthemes were: ‘going with the flow’; active participation in activities that led to new experiences (‘actually doing it’ – seeing the fun side); the importance of others in the experience of fun (‘you do games in unity’ – ‘it's not as fun on your own’). Conclusion: Children have fun when engaged in interactive and varied activities with opportunity for individual feedback and improvement. When designing childhood obesity programmes, conditions that optimise children's experience of fun should be emphasised over didactic and risk-heavy information pertaining to childhood obesity.

Caracterização fisiológica e nutritiva da clorose férrica em citrinos

Tese dout., Ciências Agrárias, Produção Vegetal, Unidade de Ciências e Tecnologias Agrárias, Universidade do Algarve, 2000

Effect of nitrogen and potassium fertilisation on vegetative growth and flowering of mature carob trees (Ceratonia siliqua): variations in leaf area index and water use indices

This work aimed to assess how potassium (K) and nitrogen (N) fertilisation may affect the use of precipitation in terms of vegetative and flowering response of 15-year-old carob trees during a 3-year experiment. A field trial was conducted in 1997, 1998 and 1999 in Algarve (Southern Portugal) in a calcareous soil. Four fertilisation treatments were tested: no fertiliser (control); 0.8 kg N/tree (N treatment); 1 kg K 2 O/tree (K treatment) and 0.8 kg N/tree plus 1 kg K 2 O/tree (NK treatment). No irrigation was applied during the experimental period. Branch length increments were measured every month throughout the growing season and inflorescence number was registered once per year. There was a strong seasonal effect on vegetative growth, since low levels of precipitation (115 mm) during October 1998–March 1999 suppressed the increment in branch length. N supplied to the trees (N and NK treatments) tended to increase water use indices in terms of vegetative growth. No response to K alone was observed in trees fertilised only with K. The number of inflorescences increased throughout the experimental period, particularly for N and NK treatments, and a reduction of the precipitation amount during April, May and June, may also enhance flowering. This knowledge could be important when making decisions concerning fertilisation under dry conditions. The results reported here indicate that tree growth (expressed as the branch growth) and flower production under dry-farming conditions, may be achieved by applying 0.8 kg of N (as ammonium nitrate) per tree during the growing season. However, N uptake and use depends on soil water availability.

Antioxidant properties and phenol content of Portulaca oleracea L. leaf, stems and flowers infusions: health benefits

Dissertação de mest., Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Univ. do Algarve, 2011

Estimating depth and explosive charge weight for an extremely shallow underwater explosion of the ROKS Cheonan sinking in the Yellow Sea

We estimated the detonation depth and net explosive weight for a very shallow underwater explosion using cutoff frequencies and spectral analysis. With detonation depth and a bubble pulse the net explosive weight for a shallow underwater explosion could simply be determined. The ray trace modeling confirms the detonation depth as a source of the hydroacoustic wave propagation in a shallow channel. We found cutoff frequencies of the reflection off the ocean bottom to be 8.5 Hz, 25 Hz, and 43 Hz while the cutoff frequency of the reflection off the free surface to be 45 Hz including 1.01 Hz for the bubble pulse, and also found the cutoff frequency of surface reflection to well fit the ray-trace modeling. We also attempted to corroborate our findings using a 3D bubble shape modeling and boundary element method. Our findings led us to the net explosive weight of the underwater explosion offshore of Baengnyeong-do for the ROKS Cheonan sinking to be approximately 136 kg TNT at a depth of about 8 m within an ocean depth of around 44 m. © 2015 Elsevier B.V.

AB1258­HPR pilot study: does a mediterranean diet­based weight loss programme improve health outcomes in overweight older women with knee osteoarthritis?

Osteoarthritis (OA) is one of the leading causes of pain and disability among older adults, particularly women. Pain and mobility disability are major consequences of knee OA which and can interfere with the functional autonomy of elderly and thus, making it difficult to perform activities of daily living. Evidence suggests that obesity is strongly linked to knee OA and that non­pharmacological therapy should be based on physical activity and weight loss in case of overweight and obesity. A positive relationship between adherence to the Mediterranean diet and health outcomes has been widely discussed in scientific literature, including its potential benefits in weight loss.

The attractive female body weight and female body dissatisfaction in 26 countries across 10 world regions: results of the International Body Project I

This study reports results from the first International Body Project (IBP-I), which surveyed 7,434 individuals in 10 major world regions about body weight ideals and body dissatisfaction. Participants completed the female Contour Drawing Figure Rating Scale (CDFRS) and self-reported their exposure to Western and local media. Results indicated there were significant cross-regional differences in the ideal female figure and body dissatisfaction, but effect sizes were small across high-socioeconomic-status (SES) sites. Within cultures, heavier bodies were preferred in low-SES sites compared to high-SES sites in Malaysia and South Africa (ds = 1.94-2.49) but not in Austria. Participant age, body mass index (BMI), and Western media exposure predicted body weight ideals. BMI and Western media exposure predicted body dissatisfaction among women. Our results show that body dissatisfaction and desire for thinness is commonplace in high-SES settings across world regions, highlighting the need for international attention to this problem.

Effects of targeted delivery of propionate to the human colon on appetite regulation, body weight maintenance and adiposity in overweight adults.

OBJECTIVE: The colonic microbiota ferment dietary fibres, producing short chain fatty acids. Recent evidence suggests that the short chain fatty acid propionate may play an important role in appetite regulation. We hypothesised that colonic delivery of propionate would increase peptide YY (PYY) and glucagon like peptide-1 (GLP-1) secretion in humans, and reduce energy intake and weight gain in overweight adults. DESIGN: To investigate whether propionate promotes PYY and GLP-1 secretion, a primary cultured human colonic cell model was developed. To deliver propionate specifically to the colon, we developed a novel inulin-propionate ester. An acute randomised, controlled cross-over study was used to assess the effects of this inulin-propionate ester on energy intake and plasma PYY and GLP-1 concentrations. The long-term effects of inulin-propionate ester on weight gain were subsequently assessed in a randomised, controlled 24-week study involving 60 overweight adults. RESULTS: Propionate significantly stimulated the release of PYY and GLP-1 from human colonic cells. Acute ingestion of 10 g inulin-propionate ester significantly increased postprandial plasma PYY and GLP-1 and reduced energy intake. Over 24 weeks, 10 g/day inulin-propionate ester supplementation significantly reduced weight gain, intra-abdominal adipose tissue distribution, intrahepatocellular lipid content and prevented the deterioration in insulin sensitivity observed in the inulin-control group. CONCLUSIONS: These data demonstrate for the first time that increasing colonic propionate prevents weight gain in overweight adult humans.