The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogen... more The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogenous in Cobrançosa table olives from northeast Portugal, were assessed in terms of functional properties and health benefits. Fourteen lactic acid bacteria strains were compared with Lacticaseibacillus casei from a commercial brand of probiotic yoghurt and L. pentosus B281 from Greek probiotic table olives, in attempts to select strains with higher probiotic performances than those references. For functional properties, the i53 and i106 strains, respectively, exhibited: 22.2 ± 2.2% and 23.0 ± 2.2% for Caco-2 cell adhesion capacity; 21.6 ± 7.8% and 21.5 ± 1.4% for hydrophobicity; 93.0 ± 3.0% and 88.5 ± 4.5% for autoaggregation ability by 24 h of incubation; and ability to co-aggregate with selected pathogens—from 29 to 40% to Gram+ (e.g., Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212); and from 16 to 44% for Gram− (e.g., Escherichia coli ATCC 25922 and Salmonella en...
Meat and fish are introduced into the diet as a source of protein, but these muscle foods present... more Meat and fish are introduced into the diet as a source of protein, but these muscle foods present different fatty acid (FA) compositions and different lipid stabilities. Fatty fish is expected to oxidize due to its higher content of polyunsaturated FA (PUFA), whereas the higher heme-Fe content of red meat will also affect lipid stability. Combining other food ingredients within a meal also influences lipid oxidation, which will not stop after meals intake. This is due to the acidic environment of the stomach together with the presence of metallic ions, a process that is scarcely understood. The goal of this study was to evaluate the oxidation of fatty fish vs. meat meal diets under in vitro standardized semi-dynamic gastric conditions and FA release from the stomach to the duodenum. Meal diets composed by 25% beef meal (BM) or fatty fish meal (FM), 25% fried potatoes, and 50% sugar soft drink were prepared. Proximate composition, FA and amino acid profiles, and meals quality indices...
Limited information exists on the bioaccessibility and intestinal uptake of essential minerals fr... more Limited information exists on the bioaccessibility and intestinal uptake of essential minerals from different types of beans cooked through different cooking methods. This study aimed to estimate the in vitro bioaccessibility and intestinal uptake of the essential minerals K, Ca, Mg, Fe, Zn, Mn, Cu and Mo from four different types of beans (kidney, black, white and cowpea) cooked under different conditions (home-cooked and industrially processed canned product). Results showed that the bioaccessibility and uptake of most essential minerals is higher in canned beans. Mn was the mineral that presented the highest bioaccessibility (43-63%), and Mo had the lowest (3-36%). The highest uptake was observed for Mg (59.9%), while the lowest was found for Fe (10.5%). Regarding the type of beans, white beans presented the highest bioaccessibility and uptake for all the analyzed essential minerals and thus its consumption is preferable from a nutritional point of view.
This study was designed to investigate whether reduced adenosine formation linked to deficits in ... more This study was designed to investigate whether reduced adenosine formation linked to deficits in extracellular ATP hydrolysis by NTPDases contributes to detrusor neuromodulatory changes associated with bladder outlet obstruction in men with benign prostatic hyperplasia (BPH). The kinetics of ATP catabolism and adenosine formation as well as the role of P1 receptor agonists on muscle tension and nerve-evoked [ 3 H]ACh release were evaluated in mucosal-denuded detrusor strips from BPH patients (n=31) and control organ donors (n=23). The neurogenic release of ATP and [ 3 H]ACh was higher (P<0.05) in detrusor strips from BPH patients. The extracellular hydrolysis of ATP and, subsequent, adenosine formation was slower (t 1/2 73 vs. 36 min, P<0.05) in BPH detrusor strips. The A 1 receptor-mediated inhibition of evoked [ 3 H]ACh release by adenosine (100 μM), NECA (1 μM), and R-PIA (0.3 μM) was enhanced in BPH bladders. Relaxation of detrusor contractions induced by acetylcholine required 30-fold higher concentrations of adenosine. Despite VAChT-positive cholinergic nerves exhibiting higher A 1 immunoreactivity in BPH bladders, the
The treatment of hypertension is of major importance to reduce the risk of cardiovascular disease... more The treatment of hypertension is of major importance to reduce the risk of cardiovascular disease, the leading cause of death worldwide. Angiotensin-converting enzyme (ACE) inhibitors are anti-hypertensive drugs associated with several side effects. Natural products, namely bioactive peptides from brewing by-products, brewers’ spent grain (BSG), and yeast (BSY), are promising alternatives since they can inhibit ACE in vitro. However, the oral intake of these peptides may modify their expected inhibitory effect owing to possible changes in active peptides’ bioavailability, which have not been assessed so far. The goal of this study was to simulate oral administration to evaluate BSG/BSY peptides’ effectiveness by submitting protein hydrolysates sequentially to simulated gastrointestinal digestion, intestinal absorption (Caco-2 cells), and liver metabolism (HepG2 cells). MTT assay was used to assess BSG/BSY protein hydrolysates safeness. The ACE-inhibitory potential of initial and fin...
Ageing beer in contact with wood is a common technological procedure that has been used for centu... more Ageing beer in contact with wood is a common technological procedure that has been used for centuries to improve colour, structure, and certain flavours. Herein, the impact of the addition of French and American oak wood to two beer styles, pale and dark, on beer phenolic composition (total phenolics, total flavonoids, and HPLC-DAD) and bioactivity (FRAP, DPPH, anti-inflammatory activity in RAW 264.7, and antiproliferative in Caco-2 cells) was assessed. Thirteen phenolics were quantified with values according to previous reports. Dark malt resulted in higher values of total phenolics, to which m-hydroxybenzoic, syringic, p-coumaric acids, and xanthohumol contributed considerably; the exception was (+)-catechin and salicylic acid, which were found to be higher in pale beers. American oak significantly increased 3,4-dihydroxyphenylacetic, vanillic, and syringic acids up to roughly 3, 2, and 10 times, respectively, when compared with French wood. FRAP and DPPH values varied between pal...
Fetal stress is known to increase susceptibility to cardiometabolic diseases and hypertension in ... more Fetal stress is known to increase susceptibility to cardiometabolic diseases and hypertension in adult age in a process known as fetal programming. This study investigated the relationship between vascular RAS, oxidative damage and remodeling in fetal programming. Six-month old Sprague-Dawley offspring from mothers that were fed ad libitum (CONTROL) or with 50% intake during the second half of gestation (maternal undernutrition, MUN) were used. qPCR or immunohistochemistry were used to obtain the expression of receptors and enzymes. Plasma levels of carbonyls were measured by spectrophotometry. In mesenteric arteries from MUN rats we detected an upregulation of ACE, ACE2, AT1 receptors and NADPH oxidase, and lower expression of AT2, Mas and MrgD receptors compared to CONTROL. Systolic and diastolic blood pressure and plasma levels of carbonyls were higher in MUN than in CONTROL. Vascular morphology evidenced an increased media/lumen ratio and adventitia/lumen ratio, and more connect...
A simple, rapid and accurate method for derivatization and quantitative HPLC analysis of organic ... more A simple, rapid and accurate method for derivatization and quantitative HPLC analysis of organic acids, was optimised in order to provide the determination of the most important acids found in Port wines and grape musts used for their elaboration. The method was based on the use of O-(4-nitrobenzyl)-N,N'-diisopropylisourea (NBDI) as derivatizing reagent. Samples were primarily treated 15 min with a strong cation-exchange resin (Dowex 50W-X8) to free the organic acids. HPLC determination was accomplished by using a RP-18 (3 µm) column, a mobile phase composed of water and acetonitrile (non-linear gradient) at a flow-rate of 1 mL min-1 , and a UV detector set at 265 nm. Sensitive and well-resolved peaks were obtained from the main organic acids (lactic, acetic, succinic, tartaric, malic and citric acids) in less than 30 min, either with Port wines or grape musts. Benzylmalonic acid was used as internal standard. Precision and recovery assays were performed with good results for all of 6 acids considered. The major differences between grape musts and Port wines were the greater overall amounts of organic acids in the former and the higher content of lactic, acetic and succinic acids in the latter.
This study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production ... more This study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production can induce resistance to azoles in Aspergillus strains and if it can induce selection of resistant species. We also tried to identify the Aspergillus species most prevalent in the vineyards. Two vineyards from northern Portugal were selected from "Vinhos Verdes" and "Douro" regions. The vineyards were divided into plots that were treated or not with penconazole (PEN). In each vineyard, air, soil, and plant samples were collected at three different times. The strains of Aspergillus spp. were isolated and identified by morphological and molecular techniques. We identified 46 Aspergillus section Nigri, eight Aspergillus fumigatus, seven Aspergillus lentulus, four Aspergillus wentii, two Aspergillus flavus, two Aspergillus terreus, one Aspergillus calidoustus, one Aspergillus westerdijkiae, one Aspergillus tamarii, and one Eurotium amstelodami. Aspergillus strains were evaluated for their susceptibility to medical azoles used in human therapy (itraconazole, posaconazole, and voriconazole) and to agricultural azoles (PEN) used in the prevention and treatment of plant diseases. The isolates showed moderate susceptibility to voriconazole. We did not observe any decrease of susceptibility to the medical azoles tested throughout the testing period in any of the treated plots, although some of the resistant species were isolated from there.
Scorpiurus (tribe Loteae) is characterised by its fruit, which is a contorted lomentum that gives... more Scorpiurus (tribe Loteae) is characterised by its fruit, which is a contorted lomentum that gives rise to the common name of «prickly caterpillar». A self re-seeding annual legume, it has potential as a forage crop due to its high nutritive values and its preference by ruminants, but has generally been neglected by researchers and farmers (Abbate et al., 2010). Native to Southern and Central Europe, the genus is clearly monophyletic having distinctive pollen morphology (Díez and Ferguson, 1996). However, specific level taxonomy remains controversial. Linnaeus (1753) recognized four species, Scorpiurus muricatus, Scorpiurus sulcatus, Scorpiurus vermiculatus and Scorpiurus subvillosus. In a study of three of these species, S. muricatus, S. sulcatus and S. subvillosus, Heyn and Raviv (1966) reported there was «no indication of any genetic crossing barriers bet
Journal of the Science of Food and Agriculture, 2004
Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal ... more Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal selection, highly recommended qualitative and productive clones have been identified. In the genus Vitis several molecular markers have been used to characterize genetic diversity at the DNA level, namely microsatellites, ISSR (inter simple sequence repeat), AFLP (amplified fragment length polymorphism), and stilbene synthase-chalcone synthase (StSy-CHS) markers. In this work a novel pure DNA extraction methodology, which occludes the extract in an agarose matrix, thus allowing the removal of the remaining hydrophilic impurities, was used. An optimized and highly reproducible procedure for the detection of differences among Vitis vinifera L clones using the StSy-CHS gene family primer combination is described. With this methodology, 21 clones and a commercial cultivar, 'Chardonnay', were evaluated with three StSy-CHS markers. It was possible to obtain 14% discrimination within Touriga Nacional (TN) clones, identifying two different groups. The polyclonal origin of TN clones was previously verified with eight microsatellite markers. The methodology described could be used for clonal selection of grapevine cultivars.
In this study, patulin levels in apples and tomatoes obtained in conventional and organic farms w... more In this study, patulin levels in apples and tomatoes obtained in conventional and organic farms with 0 and 100% of rotten area, were determined by gas chromatographyemass spectrometry (GCeMS) to assess the influence of the production system with patulin content. Additionally, mould species were isolated and identified using classic morphological techniques and a DNA barcode system based on nrITS, b-tubulin and calmodulin genes. The most frequently isolated genera were Rhizopus, Mucor, Alternaria, Cladosporium, Botrytis, Aspergillus and Penicillium. Higher patulin levels were found in samples contaminated by Penicillium expansum. Patulin levels were not significantly different in samples obtained from the two production system (conventional and organic). Among the three studied apple varieties, Golden delicious was the most contaminated with patulin followed by Reineta. Fuji apples with 0 and 100% of rotten area did not present quantifiable patulin levels. A survey for first time realized in commercialized tomato products showed that 35.7% of the samples (10 of 28) contained patulin in levels ranging from 3.22 to 47.72 mg/kg. In apple juices, patulin levels ranged from 1.86 to 45.47 mg/kg, which is below the maximum legal requirement (50 mg/kg).
The correct phylogenetic position of Lotus conimbricensis Brot. (Leguminosae, Loteae) based on nu... more The correct phylogenetic position of Lotus conimbricensis Brot. (Leguminosae, Loteae) based on nuclear ribosomal ITS sequences The nrDNA ITS sequence determined in Lotus conimbricensis in a previous phylogenetic study was unusual, in that it was almost identical to those retrieved from the morphologically distinct species L. subbiflorus. In the present study we sequenced new specimens of both species to reassess the phylogenetic position of L. conimbricensis. We conclude that the ITS sequence of L. conimbricensis used in the earlier analyses was most likely erroneous, and in fact L. conimbricensis is not closely related to L. subbiflorus. Critical reexamination of previously published data indicates that several other similar errors may exist for other Lotus species, and these should be checked before taxonomic conclusions are made.
A faster and more robust version of a previously developed method based on ion-pair extraction, a... more A faster and more robust version of a previously developed method based on ion-pair extraction, acylation with isobutylchloroformate and gas chromatography-mass spectrometry (GC-MS) analysis for determination of 4-(5-)methylimidazole (4-MeI) in soft drinks and dark beer is proposed. The performance of the method was evaluated in terms of linearity (r always > 0.998); recovery (90-101%, 3 levels); and precision (3-8%, 3 levels, n = 6). Limits of detection and quantification in the matrices studied were 0.60 mg/L and 2.2 mg/L, respectively. The optimized method was applied to a wide variety of soft drinks (brand name and generic colas, uncarbonated flavor and energy drinks) and dark beers (lager, ales trappist, ales-stout, weissbier). Overall, soft drinks presented higher amounts of 4-MeI (ranging from 37 to 613 mg/L) than those found in the dark beers (ranging from 3 to 424 mg/L), with colas presenting the highest levels. When the different colas analyzed were compared, the 4-MeI levels in generic colas were generally higher than those in brand-name colas. 4-MeI was found in only one of eight energy drinks studied. Based on available consumption patterns, consumer exposure to the maximum 4-MeI given by the soft drinks was 2.3 and 5.7 mg/kg body weight/day, in Europe and the United States, respectively.
Sensory, olfactometry (using the sums of odour intensities for each class of compounds) and chemo... more Sensory, olfactometry (using the sums of odour intensities for each class of compounds) and chemometric analyses were used to evaluate Portuguese wild hops’ sensory characteristics and the aroma that those hops impart to dry-hopped beer. CATA analysis and agglomerative hierarchical clustering was applied for the sensory characterization of 15 wild hops of Portuguese genotypes, clustering them in two groups: one more sulphurous, floral, and fruity, and another more earthy, resinous, floral, and non-citrus fruits. Two hops representative of each group were selected for the production of four dry-hopped beers using the same base beer style (Munich Helles). Beers were analysed by quantitative descriptive analyses and quantification of hop-derived key volatile compounds. Multivariate statistical treatment of the data was performed. Results indicate significant differences (p < 0.05) in fruity, resinous, earthy, floral, and sulphurous attributes of hops, but the dry-hopped beers only h...
The effects of angiotensin converting enzyme inhibition and angiotensin II receptor blockade on t... more The effects of angiotensin converting enzyme inhibition and angiotensin II receptor blockade on the development of cardiac hypertrophy and myocardial insulin-like growth factor I (IGF-I) in volume overload were studied in male Wistar rats with aorto-caval fistulas (ACF). Rats were treated with ramipril (RAM, 3 mg kg(-1) day(-1)) for 4-20 days or losartan (LOS, 10 mg kg(-1) day(-1)) for 2-7 days. Myocardial IGF-I and IGF-I receptor (IGF-I-R) mRNA were determined by solution hybridization. ACF caused hypertrophy of left (LV) and right ventricles (RV). Hypertrophy appeared on day 2 and reached maximal values of +60% in LV and +75% in RV at day 12. Systolic blood pressure was initially reduced 15% but recovered by day 12. RAM abolished the recovery of blood pressure. Furthermore, RAM attenuated RV hypertrophy by 17% on day 7 and on day 20, RV weights were close to values found in controls. Beginning on day 9, RAM reduced LV weight back to control levels in parallel to blood pressure. In contrast, LOS affected neither RV nor LV hypertrophy. RV IGF-I mRNA increased 60-100% on day 7 alone in RV in ACF. RAM potentiated the increase in RV IGF-I to +400% and induced an increase in RV IGF-I-R mRNA on day 7 (+90%) in ACF. LOS did not affect RV IGF-I. Development of cardiac hypertrophy in ACF seemed independent of angiotensin II. RV hypertrophy was associated with activation of IGF-I independent of the renin-angiotensin system. IGF-I was further potentiated when development of hypertrophy was attenuated, possibly indicative of a greater urge for compensational growth in a relatively thinner and more volume-distended chamber.
Genetic diversity within Iberian populations of Ornithopus pinnatus, O. compressus, O. sativus an... more Genetic diversity within Iberian populations of Ornithopus pinnatus, O. compressus, O. sativus and Biserrula pelecinus were assessed using ITS1 and ITS2 DNA sequences from sixty four specimens, and a phylogeny between Ornithopus species was estimated. Generally within-species variation was low, particularly within Ornithopus. The Mediterranean species of Ornithopus form a sister clade relative to the South American O. micranthopus. The sometimes considered a full species, O. sativus isthmocarpus, was not distinct from O. sativus. Between some species there is limited genetic divergence using these markers, although the situation of O. perpusillus requires additional specimens to be examined before firm conclusions can be drawn.
Journal of The Science of Food and Agriculture, 2004
Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal ... more Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal selection, highly recommended qualitative and productive clones have been identified. In the genus Vitis several molecular markers have been used to characterize genetic diversity at the DNA level, namely microsatellites, ISSR (inter simple sequence repeat), AFLP (amplified fragment length polymorphism), and stilbene synthase–chalcone synthase (StSy–CHS) markers.In this work a novel pure DNA extraction methodology, which occludes the extract in an agarose matrix, thus allowing the removal of the remaining hydrophilic impurities, was used. An optimized and highly reproducible procedure for the detection of differences among Vitis vinifera L clones using the StSy–CHS gene family primer combination is described. With this methodology, 21 clones and a commercial cultivar, ‘Chardonnay’, were evaluated with three StSy–CHS markers. It was possible to obtain 14% discrimination within Touriga Nac...
The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogen... more The probiotic features of Lactiplantibacillus (L.) pentosus and L. paraplantarum strains, endogenous in Cobrançosa table olives from northeast Portugal, were assessed in terms of functional properties and health benefits. Fourteen lactic acid bacteria strains were compared with Lacticaseibacillus casei from a commercial brand of probiotic yoghurt and L. pentosus B281 from Greek probiotic table olives, in attempts to select strains with higher probiotic performances than those references. For functional properties, the i53 and i106 strains, respectively, exhibited: 22.2 ± 2.2% and 23.0 ± 2.2% for Caco-2 cell adhesion capacity; 21.6 ± 7.8% and 21.5 ± 1.4% for hydrophobicity; 93.0 ± 3.0% and 88.5 ± 4.5% for autoaggregation ability by 24 h of incubation; and ability to co-aggregate with selected pathogens—from 29 to 40% to Gram+ (e.g., Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212); and from 16 to 44% for Gram− (e.g., Escherichia coli ATCC 25922 and Salmonella en...
Meat and fish are introduced into the diet as a source of protein, but these muscle foods present... more Meat and fish are introduced into the diet as a source of protein, but these muscle foods present different fatty acid (FA) compositions and different lipid stabilities. Fatty fish is expected to oxidize due to its higher content of polyunsaturated FA (PUFA), whereas the higher heme-Fe content of red meat will also affect lipid stability. Combining other food ingredients within a meal also influences lipid oxidation, which will not stop after meals intake. This is due to the acidic environment of the stomach together with the presence of metallic ions, a process that is scarcely understood. The goal of this study was to evaluate the oxidation of fatty fish vs. meat meal diets under in vitro standardized semi-dynamic gastric conditions and FA release from the stomach to the duodenum. Meal diets composed by 25% beef meal (BM) or fatty fish meal (FM), 25% fried potatoes, and 50% sugar soft drink were prepared. Proximate composition, FA and amino acid profiles, and meals quality indices...
Limited information exists on the bioaccessibility and intestinal uptake of essential minerals fr... more Limited information exists on the bioaccessibility and intestinal uptake of essential minerals from different types of beans cooked through different cooking methods. This study aimed to estimate the in vitro bioaccessibility and intestinal uptake of the essential minerals K, Ca, Mg, Fe, Zn, Mn, Cu and Mo from four different types of beans (kidney, black, white and cowpea) cooked under different conditions (home-cooked and industrially processed canned product). Results showed that the bioaccessibility and uptake of most essential minerals is higher in canned beans. Mn was the mineral that presented the highest bioaccessibility (43-63%), and Mo had the lowest (3-36%). The highest uptake was observed for Mg (59.9%), while the lowest was found for Fe (10.5%). Regarding the type of beans, white beans presented the highest bioaccessibility and uptake for all the analyzed essential minerals and thus its consumption is preferable from a nutritional point of view.
This study was designed to investigate whether reduced adenosine formation linked to deficits in ... more This study was designed to investigate whether reduced adenosine formation linked to deficits in extracellular ATP hydrolysis by NTPDases contributes to detrusor neuromodulatory changes associated with bladder outlet obstruction in men with benign prostatic hyperplasia (BPH). The kinetics of ATP catabolism and adenosine formation as well as the role of P1 receptor agonists on muscle tension and nerve-evoked [ 3 H]ACh release were evaluated in mucosal-denuded detrusor strips from BPH patients (n=31) and control organ donors (n=23). The neurogenic release of ATP and [ 3 H]ACh was higher (P<0.05) in detrusor strips from BPH patients. The extracellular hydrolysis of ATP and, subsequent, adenosine formation was slower (t 1/2 73 vs. 36 min, P<0.05) in BPH detrusor strips. The A 1 receptor-mediated inhibition of evoked [ 3 H]ACh release by adenosine (100 μM), NECA (1 μM), and R-PIA (0.3 μM) was enhanced in BPH bladders. Relaxation of detrusor contractions induced by acetylcholine required 30-fold higher concentrations of adenosine. Despite VAChT-positive cholinergic nerves exhibiting higher A 1 immunoreactivity in BPH bladders, the
The treatment of hypertension is of major importance to reduce the risk of cardiovascular disease... more The treatment of hypertension is of major importance to reduce the risk of cardiovascular disease, the leading cause of death worldwide. Angiotensin-converting enzyme (ACE) inhibitors are anti-hypertensive drugs associated with several side effects. Natural products, namely bioactive peptides from brewing by-products, brewers’ spent grain (BSG), and yeast (BSY), are promising alternatives since they can inhibit ACE in vitro. However, the oral intake of these peptides may modify their expected inhibitory effect owing to possible changes in active peptides’ bioavailability, which have not been assessed so far. The goal of this study was to simulate oral administration to evaluate BSG/BSY peptides’ effectiveness by submitting protein hydrolysates sequentially to simulated gastrointestinal digestion, intestinal absorption (Caco-2 cells), and liver metabolism (HepG2 cells). MTT assay was used to assess BSG/BSY protein hydrolysates safeness. The ACE-inhibitory potential of initial and fin...
Ageing beer in contact with wood is a common technological procedure that has been used for centu... more Ageing beer in contact with wood is a common technological procedure that has been used for centuries to improve colour, structure, and certain flavours. Herein, the impact of the addition of French and American oak wood to two beer styles, pale and dark, on beer phenolic composition (total phenolics, total flavonoids, and HPLC-DAD) and bioactivity (FRAP, DPPH, anti-inflammatory activity in RAW 264.7, and antiproliferative in Caco-2 cells) was assessed. Thirteen phenolics were quantified with values according to previous reports. Dark malt resulted in higher values of total phenolics, to which m-hydroxybenzoic, syringic, p-coumaric acids, and xanthohumol contributed considerably; the exception was (+)-catechin and salicylic acid, which were found to be higher in pale beers. American oak significantly increased 3,4-dihydroxyphenylacetic, vanillic, and syringic acids up to roughly 3, 2, and 10 times, respectively, when compared with French wood. FRAP and DPPH values varied between pal...
Fetal stress is known to increase susceptibility to cardiometabolic diseases and hypertension in ... more Fetal stress is known to increase susceptibility to cardiometabolic diseases and hypertension in adult age in a process known as fetal programming. This study investigated the relationship between vascular RAS, oxidative damage and remodeling in fetal programming. Six-month old Sprague-Dawley offspring from mothers that were fed ad libitum (CONTROL) or with 50% intake during the second half of gestation (maternal undernutrition, MUN) were used. qPCR or immunohistochemistry were used to obtain the expression of receptors and enzymes. Plasma levels of carbonyls were measured by spectrophotometry. In mesenteric arteries from MUN rats we detected an upregulation of ACE, ACE2, AT1 receptors and NADPH oxidase, and lower expression of AT2, Mas and MrgD receptors compared to CONTROL. Systolic and diastolic blood pressure and plasma levels of carbonyls were higher in MUN than in CONTROL. Vascular morphology evidenced an increased media/lumen ratio and adventitia/lumen ratio, and more connect...
A simple, rapid and accurate method for derivatization and quantitative HPLC analysis of organic ... more A simple, rapid and accurate method for derivatization and quantitative HPLC analysis of organic acids, was optimised in order to provide the determination of the most important acids found in Port wines and grape musts used for their elaboration. The method was based on the use of O-(4-nitrobenzyl)-N,N'-diisopropylisourea (NBDI) as derivatizing reagent. Samples were primarily treated 15 min with a strong cation-exchange resin (Dowex 50W-X8) to free the organic acids. HPLC determination was accomplished by using a RP-18 (3 µm) column, a mobile phase composed of water and acetonitrile (non-linear gradient) at a flow-rate of 1 mL min-1 , and a UV detector set at 265 nm. Sensitive and well-resolved peaks were obtained from the main organic acids (lactic, acetic, succinic, tartaric, malic and citric acids) in less than 30 min, either with Port wines or grape musts. Benzylmalonic acid was used as internal standard. Precision and recovery assays were performed with good results for all of 6 acids considered. The major differences between grape musts and Port wines were the greater overall amounts of organic acids in the former and the higher content of lactic, acetic and succinic acids in the latter.
This study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production ... more This study assessed if the use of sterol demethylase inhibitor fungicides in vineyard production can induce resistance to azoles in Aspergillus strains and if it can induce selection of resistant species. We also tried to identify the Aspergillus species most prevalent in the vineyards. Two vineyards from northern Portugal were selected from "Vinhos Verdes" and "Douro" regions. The vineyards were divided into plots that were treated or not with penconazole (PEN). In each vineyard, air, soil, and plant samples were collected at three different times. The strains of Aspergillus spp. were isolated and identified by morphological and molecular techniques. We identified 46 Aspergillus section Nigri, eight Aspergillus fumigatus, seven Aspergillus lentulus, four Aspergillus wentii, two Aspergillus flavus, two Aspergillus terreus, one Aspergillus calidoustus, one Aspergillus westerdijkiae, one Aspergillus tamarii, and one Eurotium amstelodami. Aspergillus strains were evaluated for their susceptibility to medical azoles used in human therapy (itraconazole, posaconazole, and voriconazole) and to agricultural azoles (PEN) used in the prevention and treatment of plant diseases. The isolates showed moderate susceptibility to voriconazole. We did not observe any decrease of susceptibility to the medical azoles tested throughout the testing period in any of the treated plots, although some of the resistant species were isolated from there.
Scorpiurus (tribe Loteae) is characterised by its fruit, which is a contorted lomentum that gives... more Scorpiurus (tribe Loteae) is characterised by its fruit, which is a contorted lomentum that gives rise to the common name of «prickly caterpillar». A self re-seeding annual legume, it has potential as a forage crop due to its high nutritive values and its preference by ruminants, but has generally been neglected by researchers and farmers (Abbate et al., 2010). Native to Southern and Central Europe, the genus is clearly monophyletic having distinctive pollen morphology (Díez and Ferguson, 1996). However, specific level taxonomy remains controversial. Linnaeus (1753) recognized four species, Scorpiurus muricatus, Scorpiurus sulcatus, Scorpiurus vermiculatus and Scorpiurus subvillosus. In a study of three of these species, S. muricatus, S. sulcatus and S. subvillosus, Heyn and Raviv (1966) reported there was «no indication of any genetic crossing barriers bet
Journal of the Science of Food and Agriculture, 2004
Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal ... more Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal selection, highly recommended qualitative and productive clones have been identified. In the genus Vitis several molecular markers have been used to characterize genetic diversity at the DNA level, namely microsatellites, ISSR (inter simple sequence repeat), AFLP (amplified fragment length polymorphism), and stilbene synthase-chalcone synthase (StSy-CHS) markers. In this work a novel pure DNA extraction methodology, which occludes the extract in an agarose matrix, thus allowing the removal of the remaining hydrophilic impurities, was used. An optimized and highly reproducible procedure for the detection of differences among Vitis vinifera L clones using the StSy-CHS gene family primer combination is described. With this methodology, 21 clones and a commercial cultivar, 'Chardonnay', were evaluated with three StSy-CHS markers. It was possible to obtain 14% discrimination within Touriga Nacional (TN) clones, identifying two different groups. The polyclonal origin of TN clones was previously verified with eight microsatellite markers. The methodology described could be used for clonal selection of grapevine cultivars.
In this study, patulin levels in apples and tomatoes obtained in conventional and organic farms w... more In this study, patulin levels in apples and tomatoes obtained in conventional and organic farms with 0 and 100% of rotten area, were determined by gas chromatographyemass spectrometry (GCeMS) to assess the influence of the production system with patulin content. Additionally, mould species were isolated and identified using classic morphological techniques and a DNA barcode system based on nrITS, b-tubulin and calmodulin genes. The most frequently isolated genera were Rhizopus, Mucor, Alternaria, Cladosporium, Botrytis, Aspergillus and Penicillium. Higher patulin levels were found in samples contaminated by Penicillium expansum. Patulin levels were not significantly different in samples obtained from the two production system (conventional and organic). Among the three studied apple varieties, Golden delicious was the most contaminated with patulin followed by Reineta. Fuji apples with 0 and 100% of rotten area did not present quantifiable patulin levels. A survey for first time realized in commercialized tomato products showed that 35.7% of the samples (10 of 28) contained patulin in levels ranging from 3.22 to 47.72 mg/kg. In apple juices, patulin levels ranged from 1.86 to 45.47 mg/kg, which is below the maximum legal requirement (50 mg/kg).
The correct phylogenetic position of Lotus conimbricensis Brot. (Leguminosae, Loteae) based on nu... more The correct phylogenetic position of Lotus conimbricensis Brot. (Leguminosae, Loteae) based on nuclear ribosomal ITS sequences The nrDNA ITS sequence determined in Lotus conimbricensis in a previous phylogenetic study was unusual, in that it was almost identical to those retrieved from the morphologically distinct species L. subbiflorus. In the present study we sequenced new specimens of both species to reassess the phylogenetic position of L. conimbricensis. We conclude that the ITS sequence of L. conimbricensis used in the earlier analyses was most likely erroneous, and in fact L. conimbricensis is not closely related to L. subbiflorus. Critical reexamination of previously published data indicates that several other similar errors may exist for other Lotus species, and these should be checked before taxonomic conclusions are made.
A faster and more robust version of a previously developed method based on ion-pair extraction, a... more A faster and more robust version of a previously developed method based on ion-pair extraction, acylation with isobutylchloroformate and gas chromatography-mass spectrometry (GC-MS) analysis for determination of 4-(5-)methylimidazole (4-MeI) in soft drinks and dark beer is proposed. The performance of the method was evaluated in terms of linearity (r always > 0.998); recovery (90-101%, 3 levels); and precision (3-8%, 3 levels, n = 6). Limits of detection and quantification in the matrices studied were 0.60 mg/L and 2.2 mg/L, respectively. The optimized method was applied to a wide variety of soft drinks (brand name and generic colas, uncarbonated flavor and energy drinks) and dark beers (lager, ales trappist, ales-stout, weissbier). Overall, soft drinks presented higher amounts of 4-MeI (ranging from 37 to 613 mg/L) than those found in the dark beers (ranging from 3 to 424 mg/L), with colas presenting the highest levels. When the different colas analyzed were compared, the 4-MeI levels in generic colas were generally higher than those in brand-name colas. 4-MeI was found in only one of eight energy drinks studied. Based on available consumption patterns, consumer exposure to the maximum 4-MeI given by the soft drinks was 2.3 and 5.7 mg/kg body weight/day, in Europe and the United States, respectively.
Sensory, olfactometry (using the sums of odour intensities for each class of compounds) and chemo... more Sensory, olfactometry (using the sums of odour intensities for each class of compounds) and chemometric analyses were used to evaluate Portuguese wild hops’ sensory characteristics and the aroma that those hops impart to dry-hopped beer. CATA analysis and agglomerative hierarchical clustering was applied for the sensory characterization of 15 wild hops of Portuguese genotypes, clustering them in two groups: one more sulphurous, floral, and fruity, and another more earthy, resinous, floral, and non-citrus fruits. Two hops representative of each group were selected for the production of four dry-hopped beers using the same base beer style (Munich Helles). Beers were analysed by quantitative descriptive analyses and quantification of hop-derived key volatile compounds. Multivariate statistical treatment of the data was performed. Results indicate significant differences (p < 0.05) in fruity, resinous, earthy, floral, and sulphurous attributes of hops, but the dry-hopped beers only h...
The effects of angiotensin converting enzyme inhibition and angiotensin II receptor blockade on t... more The effects of angiotensin converting enzyme inhibition and angiotensin II receptor blockade on the development of cardiac hypertrophy and myocardial insulin-like growth factor I (IGF-I) in volume overload were studied in male Wistar rats with aorto-caval fistulas (ACF). Rats were treated with ramipril (RAM, 3 mg kg(-1) day(-1)) for 4-20 days or losartan (LOS, 10 mg kg(-1) day(-1)) for 2-7 days. Myocardial IGF-I and IGF-I receptor (IGF-I-R) mRNA were determined by solution hybridization. ACF caused hypertrophy of left (LV) and right ventricles (RV). Hypertrophy appeared on day 2 and reached maximal values of +60% in LV and +75% in RV at day 12. Systolic blood pressure was initially reduced 15% but recovered by day 12. RAM abolished the recovery of blood pressure. Furthermore, RAM attenuated RV hypertrophy by 17% on day 7 and on day 20, RV weights were close to values found in controls. Beginning on day 9, RAM reduced LV weight back to control levels in parallel to blood pressure. In contrast, LOS affected neither RV nor LV hypertrophy. RV IGF-I mRNA increased 60-100% on day 7 alone in RV in ACF. RAM potentiated the increase in RV IGF-I to +400% and induced an increase in RV IGF-I-R mRNA on day 7 (+90%) in ACF. LOS did not affect RV IGF-I. Development of cardiac hypertrophy in ACF seemed independent of angiotensin II. RV hypertrophy was associated with activation of IGF-I independent of the renin-angiotensin system. IGF-I was further potentiated when development of hypertrophy was attenuated, possibly indicative of a greater urge for compensational growth in a relatively thinner and more volume-distended chamber.
Genetic diversity within Iberian populations of Ornithopus pinnatus, O. compressus, O. sativus an... more Genetic diversity within Iberian populations of Ornithopus pinnatus, O. compressus, O. sativus and Biserrula pelecinus were assessed using ITS1 and ITS2 DNA sequences from sixty four specimens, and a phylogeny between Ornithopus species was estimated. Generally within-species variation was low, particularly within Ornithopus. The Mediterranean species of Ornithopus form a sister clade relative to the South American O. micranthopus. The sometimes considered a full species, O. sativus isthmocarpus, was not distinct from O. sativus. Between some species there is limited genetic divergence using these markers, although the situation of O. perpusillus requires additional specimens to be examined before firm conclusions can be drawn.
Journal of The Science of Food and Agriculture, 2004
Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal ... more Touriga Nacional is the most popular Portuguese grapevine cultivar. After twenty years of clonal selection, highly recommended qualitative and productive clones have been identified. In the genus Vitis several molecular markers have been used to characterize genetic diversity at the DNA level, namely microsatellites, ISSR (inter simple sequence repeat), AFLP (amplified fragment length polymorphism), and stilbene synthase–chalcone synthase (StSy–CHS) markers.In this work a novel pure DNA extraction methodology, which occludes the extract in an agarose matrix, thus allowing the removal of the remaining hydrophilic impurities, was used. An optimized and highly reproducible procedure for the detection of differences among Vitis vinifera L clones using the StSy–CHS gene family primer combination is described. With this methodology, 21 clones and a commercial cultivar, ‘Chardonnay’, were evaluated with three StSy–CHS markers. It was possible to obtain 14% discrimination within Touriga Nac...
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