Olive tree (Olea europaea L.) diversity in traditional small farms of Ficalho, Portugal

The genetic diversity of “Gama” and “Bico de Corvo”, local cultivars of olive tree (Olea europaea) from seven traditional orchards of Ficalho (Alentejo region, Portugal), was studied to characterize the local diversity and assess the level of on farm diversity. Two different analytical systems were used: endocarp morphological characteristics and genetic analysis by microsatellite markers (Simple Sequence Repeats or SSR). The seven screened loci were polymorphic and allowed the identification of 23 distinct SSR profiles within the 27 trees analyzed. A total of 52 different alleles were scored, with an average of 7.43 alleles/SSR locus, and considerable genetic diversity was found. Neighbor-Joining algorithm cluster analysis and principal co-ordinate analysis (PCoA) allowed for the identification of the genetic relationships between several accessions. The 27 Olea accessions were clearly separated into three different groups. SSR analysis was more precise than endocarp characterization in the classification of genetic diversity among the olive tree cultivars. The study shows reasonable olive tree diversity in Ficalho, indicating that these traditional orchards are important reservoirs of old minor cultivars and incubators of new genotypesinfo:eu-repo/semantics/publishedVersio

Genetic Diversity and Population Structure of Wild Beets (Beta spp.) from the Western Iberian Peninsula and the Azores and Madeira Islands

In this work, using simple sequence repeat (SSR) markers, we present new insights into the genetic diversity, differentiation, and structure of Beta vulgaris subsp. maritima of western Iberia and the Azores and Madeira islands and of B. macrocarpa from southern Portugal. B. macrocarpa occurs only in southern Portugal and frequently in sympatry with B. vulgaris subsp. maritima, showing genetic introgression. B. macrocarpa has a better-defined structure than B. vulgaris subsp. maritima, which has a high degree of admixture. A great differentiation (FST ranging from 0.277 to 0.184) was observed among the northern populations of B. vulgaris subsp. maritima. In contrast, only a small differentiation (FST ranging from 0.000 to 0.026) was detected among the southern B. vulgaris subsp. maritima populations. The inland B. vulgaris subsp. maritima populations (“RIO” and “VMT”) are distinct from each other, which also occurs with the two islands’ populations (“MAD” and “AZO”). The existence of two distinct Atlantic Sea currents can explain the fact that Madeira is related to the southern populations, while the Azores is related to the northern populations. We consider that understanding the relationships existing within Beta spp. is key to future genetic studies and for the establishment of conservation measures. Our results show that the southern coastal areas of Portugal should be considered as a potential site for in situ conservation of the beet wild relatives. Special attention is needed in what concerns B. macrocarpa because this is a rare species that also occurs in a sympatric relationship with B. vulgaris subsp. maritimainfo:eu-repo/semantics/publishedVersio

Uzgoj bakterije Methylobacterium organophilum u metanolu za istodobnu proizvodnju proteina i korisnih metabolita

Research background. This study aims to monitor the growth of the methylotrophic bacteria Methylobacterium organophilum in a culture medium with methanol as a carbon source and to verify the production of unicellular proteins and other biomolecules, such as carotenoids, exopolysaccharides and polyhydroxyalkanoates, making them more attractive as animal feed. Experimental approach. Bacterial growth was studied in shake flasks using different carbon/nitrogen (C:N) ratios to determine their best ratio for achieving the highest volumetric productivity of cells and substrate consumption rate. This optimal parameter was further used in a fed-batch operating bioreactor system to define the kinetic profile of cell growth. Methanol consumption was measured by HPLC analysis and the extracted pigments were analyzed by liquid chromatography/mass spectrometry. Chemical composition and rheological properties of the produced exopolysaccharides were also determined. Results and conclusions. The best experimental parameters were verified using an initial methanol concentration of 7 g/L in the culture medium. The same initial substrate concentration was used in the fed-batch operation and after 60 h of cultivation 5 g/L of biomass were obtained. The accumulation of carotenoids associated with cell growth was monitored, reaching a concentration of 1.6 mg/L at the end of the process. These pigments were then analyzed and characterized as a set of xanthophylls (oxidized carotenoids). In addition, two other product types were identified during the fed-batch operation: exopolysaccharides, which reached a concentration of 8.9 g/L at the end of the cultivation, and an intracellular granular structure that was detected by transmission electron microscopy (TEM), suggesting the accumulation of polyhydroxyalkanoate (PHA), most likely polyhydroxybutyrate. Novelty and scientific contribution. Methylobacterium organophilum demonstrated a unique ability to produce compounds of commercial interest. The distinct metabolic diversity of this bacterium makes room for its use in biorefineries.Pozadina istraživanja. Svrha je ovoga istraživanja bila pratiti rast metilotrofne bakterije Methylobacterium organophilum u podlozi s metanolom kao izvorom ugljika, te potvrditi sposobnost tog jednostaničnog organizma da proizvodi proteine i druge biomolekule, kao što su karotenoidi, egzopolisaharidi i polihidroksialkanoati, a koji se mogu upotrijebiti u proizvodnji stočne hrane. Eksperimentalni pristup. Rast bakterije ispitan je na tresilici pri različitim omjerima ugljika i dušika (C:N), radi utvrđivanja njihovog optimalnog omjera za postizanje najveće volumetrijske produktivnosti stanica i najboljeg iskorištenja supstrata. Zatim je pomoću tog optimalnog omjera hranjiva određen kinetički profil rasta stanica u šaržnom bioreaktoru. Potrošnja metanola mjerena je metodom HPLC, a izolirani su pigmenti ispitani tekućinskom kromatografijom spregnutom s masenom spektrometrijom. Također su određeni kemijski sastav i reološka svojstva dobivenih egzopolisaharida. Rezultati i zaključci. Najbolji eksperimentalni parametri potvrđeni su pri početnoj koncentraciji metanola u podlozi od 7 g/L. Ista je početna koncentracija korištena u šaržnom bioreaktoru, gdje je nakon 60 h uzgoja dobiveno 5 g/L biomase. Tijekom uzgoja je praćena akumulacija karotenoida, čija je koncentracija pri završetku procesa bila 1,6 mg/L. Pigmenti su zatim ispitani i okarakterizirani kao ksantofili (podvrsta karotenoida). Osim toga, tijekom šaržne fermentacije identificirana su još dva proizvoda: egzopolisaharidi, čija je koncentracija na kraju uzgoja bila 8,9 g/L; te granularna struktura otkrivena pomoću transmisijskog elektronskog mikroskopa, koja upućuje na to da je u stanici došlo do nakupljanja polihidroksialkanoata (PHA), i to najvjerojatnije polihidroksibutirata. Novina i znanstveni doprinos. Bakterija Methylobacterium organophilum ima jedinstveno svojstvo proizvodnje spojeva od komercijalnog interesa. Iznimna metabolička raznolikost ove bakterije omogućuje njezinu primjenu u biorafinerijama

Aetiology of Acute Gastroenteritis in Hospitalized Children from Lisbon Area

Gastroenteritis is a major cause of morbidity and mortality worlwide. Aim: Determine the etiology of gastroenteritis in a cohort of children hospitalized Methods: From May 2011 to January 2012, stool samples were collected from children (4.2 ± 4.8 years). Viral agents (enteric and enterovirus) were detected by RT-PCR. Stool specimens were also tested for bacteria and parasites, by conventional methods. Results: From the 82 stool samples, 42(51.2%) were positive for virus, 22 (26.8%) for bacteria and 35.6% (26/73) for parasites. An association between viral infection and age<3 years (OR=4.92, p<0.01) was observed. Distribution of viral agents was NorovirusII (33.3%), Enterovirus (31%), Rotavirus (23.8%), Adenovirus (9.5%), NorovirusI (2.4%). Simultaneous detection of two viral agents was observed in seven cases (16.7%) - NorovirusII and Enterovirus was the most frequent (57.1%); association with another agent was detected in 33.3% samples. Eight of 22 samples (36.4%) were positive for Campylobacter jejuni, 5 (22.7%) for Salmonella spp, 7 (31.8%) for E. coli, although always in co-infection with another agent, and 1 (4.5%) for Shigella spp. 38.5% of the samples were positive for Cryptosporidium spp., 23.1% for Giardia sp. and 3.8% for Entamoeba histolytica. 69.2% of the parasites were co-detected with other agents. Conclusion: Results suggest that viral agents are the most common among children with acute gastroenteritis, although co-infections with bacteria and parasites are frequent. The most common viral agent associated with acute diarrhea was NorovirusII. An important percentage of cases with no infectious aetiology identified, suggesting that other emergent agents are probably implicated

Multi-step subcritical water extracts of fucus vesiculosus l. And codium tomentosum stackhouse: Composition, health-benefits and safety

PO-CI-01-0145FEDER-030240Mental health and active aging are two of the main concerns in the 21st century. To search for new neuroprotective compounds, extracts of Codium tomentosum Stackhouse and Fucus vesiculosus L. were obtained through multi-step (four step) subcritical water extraction using a temperature gradient. The safety assessment of the extracts was performed by screening pharmaceutical compounds and pesticides by UHPLC-MS/MS, and iodine and arsenic levels by ICP-MS. Although the extracts were free of pharmaceutical compounds and pesticides, the presence of arsenic and high iodine contents were found in the first two extraction steps. Thus, the health-benefits were only evaluated for the fractions obtained in steps 3 and 4 from the extraction process. These fractions were tested against five brain enzymes implicated in Alzheimer’s, Parkinson’s, and major depression etiology as well as against reactive oxygen and nitrogen species, having been observed a strong enzyme inhibition and radical scavenging activities for the step 4 fractions from both seaweed species. Regarding the variation of the chemical composition during the extraction, step 1 fractions were the richest in phenolic compounds. With the increase in temperature, Maillard reaction, caramelization and thermo-oxidation occurred, and the resulting products positively affected the antioxidant capacity and the neuroprotective effects.publishersversionpublishe

The P2X7 receptor mediates Toxoplasma gondii Control in Macrophages through canonical NLRP3 inflammasome activation and reactive oxygen species production

Toxoplasma gondii (T. gondii) is the protozoan parasite that causes toxoplasmosis, a potentially fatal disease to immunocompromised patients, and which affects approximately 30% of the world\u27s population. Previously, we showed that purinergic signaling via the P2X7 receptor contributes to T. gondii elimination in macrophages, through reactive oxygen species (ROS) production and lysosome fusion with the parasitophorous vacuole. Moreover, we demonstrated that P2X7 receptor activation promotes the production of anti-parasitic pro-inflammatory cytokines during early T. gondii infection in vivo. However, the cascade of signaling events that leads to parasite elimination via P2X7 receptor activation remained to be elucidated. Here, we investigated the cellular pathways involved in T. gondii elimination triggered by P2X7 receptor signaling, during early infection in macrophages. We focused on the potential role of the inflammasome, a protein complex that can be co-activated by the P2X7 receptor, and which is involved in the host immune defense against T. gondii infection. Using peritoneal and bone marrow-derived macrophages from knockout mice deficient for inflammasome components (NLRP3-/-, Caspase-1/11-/-, Caspase-11-/-), we show that the control of T. gondii infection via P2X7 receptor activation by extracellular ATP (eATP) depends on the canonical inflammasome effector caspase-1, but not on caspase-11 (a non-canonical inflammasome effector). Parasite elimination via P2X7 receptor and inflammasome activation was also dependent on ROS generation and pannexin-1 channel. Treatment with eATP increased IL-1β secretion from infected macrophages, and this effect was dependent on the canonical NLRP3 inflammasome. Finally, treatment with recombinant IL-1β promoted parasite elimination via mitochondrial ROS generation (as assessed using Mito-TEMPO). Together, our results support a model where P2X7 receptor activation by eATP inhibits T. gondii growth in macrophages by triggering NADPH-oxidase-dependent ROS production, and also by activating a canonical NLRP3 inflammasome, which increases IL-1β production (via caspase-1 activity), leading to mitochondrial ROS generation

Nutritional evaluation of children with chronic cholestatic disease

AbstractObjectiveTo evaluate the nutritional status of children with persistent cholestasis and to compare the anthropometric indices between children with and without liver cirrhosis and children with and without jaundice.MethodsChildren with persistent cholestasis, i.e increased direct bilirrubin or changes in the canalicular enzyme gamma‐glutamyl transferase (GGT), were included. The anthropometric measures were weight (W), height or length (H), arm circumference (AC), triceps skinfold thickness (TST), arm muscle circumference (AMC), and body mass index (BMI).ResultsNinety‐one children with cholestasis, with current median age of 12 months, were evaluated. W/age (A) and H/A indices below –2 Z‐scores were observed in 33% and 30.8% of patients, respectively. Concerning the W/H index and BMI, only 12% and 16% of patients, respectively, were below –2 Z‐scores. Regarding AC, 43.8% of 89 evaluated patients had some depletion. Observing the TST, 64% of patients had depletion, and 71.1% of the 45 evaluated patients had some degree of depletion regarding the ACM index.ConclusionEvaluation using weight in patients with chronic liver diseases may overestimate the nutritional status due to visceromegaly, subclinical edema, or ascites. Indices that correlate weight and height, such as W/H and BMI, may also not show depletion because of the chronic condition in which there are depletion of both weight and height. TST, AC, and ACM are parameters that better estimate nutritional status and should be part of the management of patients with liver diseases and cholestasis.ResumoObjetivoAvaliar a situação nutricional de crianças com colestase persistente e comparar os índices antropométricos entre crianças com e sem cirrose hepática e crianças com e sem icterícia.MétodosForam incluídas crianças com colestase persistente, ou seja, aumento da bilirrubina direta ou alterações na enzima canalicular, gamaglutamiltransferase (GGT). As medidas antropométricas foram peso, estatura ou altura, circunferência do braço (CB), espessura da prega cutânea do tríceps (TST), circunferência muscular do braço (CMB) e índice de massa corporal (IMC).ResultadosForam avaliadas 91 crianças com colestase, com idade média de 12 meses; 33% e 30,8% dos pacientes apresentaram índices P/I e A/I com escore Z abaixo de –2, respectivamente. Com relação ao índice P/A e IMC, somente 12% e 16% dos pacientes, respectivamente, apresentaram escore Z abaixo de –2. Com relação à CB, 43,8% de 89 pacientes avaliados apresentaram alguma depleção. Observando a TST, 64% dos pacientes que apresentaram depleção, 71,1% dos 45 pacientes avaliados apresentaram algum grau de depleção com relação ao índice de CMB.ConclusãoA avaliação do peso em pacientes com doenças hepáticas crônicas poderá superestimar a situação nutricional devido a visceromegalia, edema subclínico ou ascite. Os índices que correlacionam peso e altura, como P/A e IMC, também podem não mostrar depleção devido à doença crônica em que há depleção tanto do peso quanto da altura. A TST, BC e CMB são parâmetros que estimam melhor a situação nutricional e devem fazer parte de gestão de pacientes com doenças hepáticas e colestase