Cerberus is a feedback inhibitor of Nodal asymmetric signaling in the chick embryo

The TGF-beta-related molecule Nodal plays an essential and conserved role in left-right patterning of the vertebrate embryo. Previous reports have shown that the zebrafish and mouse Cerberus-related proteins Charon and Cerberus-like-2 (Cerl-2), respectively, act in the node region to prevent the Nodal signal from crossing to the right side, whereas chick Cerberus (cCer) has an unclear function in the left-side mesoderm. In this study, we investigate the transcriptional regulation and function of cCer in left-right development. By analyzing the enhancer activity of cCer 5' genomic sequences in electroporated chick embryos, we identified a cCer left-side enhancer that contains two FoxH1 and one SMAD binding site. We show that these Nodal-responsive elements are necessary and sufficient for the activation of transcription in the left-side mesoderm. In transgenic mouse embryos, cCer regulatory sequences behave as in chick embryos, suggesting that the cis-regulatory sequences of Cerberus-related genes have diverged during vertebrate evolution. Moreover, our findings from cCer overexpression and knockdown experiments indicate that cCer is a negative-feedback regulator of Nodal asymmetric signaling. We propose that cCer and mouse Cerl-2 have evolved distinct regulatory mechanisms but retained a conserved function in left-right development, which is to restrict Nodal activity to the left side of the embryo.info:eu-repo/semantics/publishedVersio

Screening Mediterranean shrubs selected by browsing goats against gastrointestinal strongyles

Ethanolic extracts from the main species selected by browsing Charnequeira goats on Mediterranean vegetation were assayed for a rapid screening of their anthelmintic potential against gastrointestinal (GI) strongyles. Shrubs selected included Olea europeae var. sylvestris (OS), Quercus coccifera (QC), Pistacia lentiscus, Rhamnus alaternus and Rhamnus lycioides (RL). Forage species included Cichorium intybus (CI) and Hedysarum coronarium (HC). Extract concentrations were chosen taking as reference the fecal total phenolics content (TP) from those goats after a five-month browsing period (8.4 mg GAE/g DM). A quantitative coproculture method was adapted using faeces obtained from naturally infected goats, not dewormed or having access to browse (reference group); larvae culture was performed in triplicate for three different TP concentrations (25, 10 and 5 mg GAE/g DM) against a distilled water control, for a 7-day incubation period at 27 °C. The number of larvae developed was counted differentiating total and live larvae per gram of faeces (LDPG). The efficacy of each extract and concentration, as percentage of reduction in LDPG against control, was determined. Data was log transformed and submitted to analysis of variance. Means were compared by Tukey test at 5% significance level.A reduction in LDPG was observed in all extracts being significant (P<0.05) for the highest concentration. The extracts’ efficacy over exogenous forms showed not only a reduction of total LDPG but, for some, also a high level of larvae mortality, resulting in reduction of live infective larvae (L3). These results indicate that some extracts, besides their ovicidal activity, have also larvicidal potential. Extracts efficacy for total LDPG was higher for QC (82%), OS (80%), HC and RL (76%) and CI (69%). Reduction of live L3 was higher for OS (90%) and HC (80%). Results obtained on these extracts potential in reducing exogenous forms, namely L3 forms of GI strongyles, is an important aspect in the control of these parasitic populations, as it results in lower pasture contamination and consequently a lower rate of reinfection of the animals. Funding obtained from project VegMedCabras – ALT20-03-0145-FEDER-000009

ValBioTecCynara – Economic Enhancement Cynara cardunculus: Natural Variability and its Biotechnological Applications

ValBioTecCynara aims to address the natural C cardunculus L genetic, molecular, morphologic, and biochemical variabilities, as a combined strategy to identify individuals with certain and specific required profiles C cardunculus L is used as a coagulant for cheesemaking where the respective use is mandatory in certain cheeses benefiting from Protected Designation of Origin ( 1 The potential of 15 different C cardunculus L natural occurring populations within Alentejo region (south of Portugal), were evaluated on flower technological properties, being three of them studied regarding chemical, rheological and sensorial characteristics within the three PDO Alentejo cheeses Évora Serpa and Nisa A significance variability on flower technological properties was found, based on milk clotting activity, gel firmness, micellar aggregation rate and proteolytic activity and it was possible to identify 5 different groups of C cardunculus L populations 2 The action of coagulant populations was specific according the cheese matrix Nevertheless, a common pattern of cheese proteolysis, rheology and sensory characteristics was verified, allowing to highlight the cheeses manufactured with two of the three C cardunculus L populations Proteolysis was more intense in early stages of ripening, influenced by cheese moisture content and raising to caseins fractions, β caseins were less susceptible to proteolysis than α S caseins with influence in cheese sensory characteristics These results have a very stimulating uses concerning specific target, such as uncertainty in the cheese manufacture, contributing to the incentive of the cheese production systems promoting the milk production of small ruminants Keywords Cynara cardunculus L proteases milk PDO Évora, Serpa, Nisa cheeses 1 Conceição C, Martins P, Alvarenga N, Dias J, Lamy E, Garrido A, et al Cynara cardunculus Use in Cheesemaking and Pharmaceutical Applications In Koca N Technological Approaches for Novel Applications in Dairy Processing InTechOpen 2018 http :://dx doi org/ 10 5772 /intechopen 76530 2 Gomes S, Belo AT, Alvarenga N, Dias J, Lage P, Pinheiro C, et al Characterization of Cynara cardunculus L flower from Alentejo as a coagulant agent for cheesemaking Int Dairy J 2019 91 178 84 This project was funded by European Regional Development Fund ( ValBioTecCynara (ALT 20 03 0145 FEDER 000038 Economic valorisation of Cardoon Cynara cardunculus study of natural variability and biotechnological applications The authors acknowledge FCT for PhD grant to Teresa Brás (SFRH/ 110969 201

Mudança de paradigma na abordagem aos parasitas gastrointestinais em pequenos ruminantes – controlo integrado com tratamento seletivo

O crescente desenvolvimento de resistências aos anti-helmínticos, com particular importância em pequenos ruminantes, tem ameaçado a sustentabilidade da produção no sector. Este desafio no controlo é exacerbado pela escassez de moléculas alternativas e pela crescente consciencialização da contaminação ambiental aquando da excreção dos compostos anti-helmínticos administrados. Neste trabalho, combinaram-se os dados preliminares recolhidos ao longo de dois projectos independentes (VegMedCabras e CistusRumen financiados pelo Fundo Europeu para o Desenvolvimento regional, Alentejo 2020), ainda em curso. Enquanto o primeiro visa promover o pastoreio da vegetação arbustiva mediterrânica de forma ecológica e sustentável por caprinos, o segundo tem por objectivo uma utilização sustentável da Esteva (Cistus ladanifer L.), em pequenos ruminantes, como ferramenta no aumento da competitividade e na redução do impacte ambiental. Em ambos, a infecção parasitária tem vindo a ser avaliada individualmente através da determinação do número de ovos eliminados por grama de fezes (OPG) e aplicada a técnica qualitativa de flutuação por Willis em todas as amostras, individualmente. Os ensaios experimentais realizados no âmbito do projecto CistusRumen incidiram sobre borregos (36 animais) e cabras adultas (18 animais), sujeitos a dietas suplementadas com diferentes concentrações de taninos ou esteva. Entre os resultados obtidos, os achados dos exames qualitativos foram os que revelaram o maior potencial dos taninos. Durante as análises coprológicas dos animais pertencentes a grupos com dietas suplementadas registou-se um fenómeno de rebentamento ou alteração da morfologia dos ovos de estrongilídeos. Os resultados preliminares recolhidos nos ensaios experimentais do CistusRumen complementam os objectivos do VegMedCabras, uma vez que a incorporação de vegetação arbustiva mediterrânica, como a Esteva, cujos taninos auxiliam a absorção proteica ao nível do intestino delgado, contribuem para uma maior capacidade de resposta do hospedeiro às infecções parasitárias. Um aumento na capacidade de resposta associada ao aparente efeito da suplementação da dieta sobre as formas exógenas diminui a necessidade do controlo pelo tratamento, reservando-o para casos específicos com tratamento selectivo do efectivo diminuindo assim a pressão de selecção sobre as resistências e diminuindo a contaminação ambiental

Screening Mediterranean shrubs selected by browsing goats against gastrointestinal strongyles

Ethanolic extracts from the main species selected by browsing Charnequeira goats on Mediterranean vegetation were assayed for a rapid screening of their anthelmintic potential against gastrointestinal (GI) strongyles. Shrubs selected included Olea europeae var. sylvestris (OS), Quercus coccifera (QC), Pistacia lentiscus, Rhamnus alaternus and Rhamnus lycioides (RL). Forage species included Cichorium intybus (CI) and Hedysarum coronarium (HC). Extract concentrations were chosen taking as reference the fecal total phenolics content (TP) from those goats after a five-month browsing period (8.4 mg GAE/g DM). A quantitative coproculture method was adapted using faeces obtained from naturally infected goats, not dewormed or having access to browse (reference group); larvae culture was performed in triplicate for three different TP concentrations (25, 10 and 5 mg GAE/g DM) against a distilled water control, for a 7-day incubation period at 27 °C. The number of larvae developed was counted differentiating total and live larvae per gram of faeces (LDPG). The efficacy of each extract and concentration, as percentage of reduction in LDPG against control, was determined. Data was log transformed and submitted to analysis of variance. Means were compared by Tukey test at 5% significance level.A reduction in LDPG was observed in all extracts being significant (P<0.05) for the highest concentration. The extracts’ efficacy over exogenous forms showed not only a reduction of total LDPG but, for some, also a high level of larvae mortality, resulting in reduction of live infective larvae (L3). These results indicate that some extracts, besides their ovicidal activity, have also larvicidal potential. Extracts efficacy for total LDPG was higher for QC (82%), OS (80%), HC and RL (76%) and CI (69%). Reduction of live L3 was higher for OS (90%) and HC (80%). Results obtained on these extracts potential in reducing exogenous forms, namely L3 forms of GI strongyles, is an important aspect in the control of these parasitic populations, as it results in lower pasture contamination and consequently a lower rate of reinfection of the animals. Funding obtained from project VegMedCabras – ALT20-03-0145-FEDER-00000

Azatioprina na doença inflamatória intestinal: fatores preditivos da resposta sustentada a longo prazo

ResumoIntroduçãoA azatioprina (AZA) é uma opção terapêutica de manutenção na doença inflamatória intestinal (DII). Contudo, os fatores que influenciam ou predizem a sua resposta são pouco conhecidos.ObjetivoAvaliar os fatores preditivos de sucesso terapêutico da AZA a longo prazo.MétodosAnálise retrospetiva de todos os doentes com DII seguidos no nosso hospital medicados com AZA (2‐2,5mg/Kg/dia) por doença corticodependente ou corticorrefratária e, na doença de Crohn (DC), por comportamento fistulizante ou após cirurgia. Registámos o tipo de doença (DC/colite ulcerosa [CU]/DII indeterminada), parâmetros clínicos, parâmetros laboratoriais (PL) – leucócitos, PCR, hemoglobina, plaquetas e VGM – antes e aos 3 meses de tratamento, bem como terapêutica concomitante com 5‐ASA e corticoide. O tratamento foi considerado eficaz quando os doentes mantinham o controlo da doença por critérios clínicos/endoscópicos, com manutenção da AZA ou com cessação da mesma após 3 meses de tratamento, e sem necessidade de escalar a terapêutica. Excluímos os doentes com intolerância à AZA nos primeiros 3 meses e os tratados concomitantemente com agentes biológicos.ResultadosSetenta e dois doentes (37 mulheres e 35 homens); idade média de 38,0±13,8 anos; 35 doentes com DC, 34 com CU e 3 com DII indeterminada. Duração média de tratamento com AZA de 35,1±30,6 meses. A AZA foi eficaz em 48 doentes (66,7%). A idade no início da AZA prediz o sucesso terapêutico (R=0,303, p=0,019). O sexo, o tipo de doença e os PL antes do tratamento não tiveram relação com a eficácia. Aos 3 meses de tratamento, os PL mostraram relação com o sucesso terapêutico a longo prazo: leucócitos (r=–0,295, p=0,013), PCR (r=–0,332, p=0,005), hemoglobina (r=0,307, p=0,010), plaquetas (r=–0,360, p=0,003) e VGM (r=0,255, p=0,047); no seu conjunto os PL predizem a eficácia (R=0,517; p=0,005). Existe associação entre a localização da CU (r=–0,381; p=0,026), o tempo de tratamento concomitante com 5‐ASA (r=0,258, p=0,029) e a suspensão de corticoides (r=0,265, p=0,04) com a eficácia do tratamento.ConclusãoO tratamento com a AZA foi eficaz na maioria dos doentes com DII. A idade avançada no início da terapêutica e os PL aos 3 meses foram preditivos de resposta sustentada da AZA.AbstractIntroductionAzathioprine (AZA) is an option for maintenance therapy in Inflammatory Bowel Disease (IBD). However, the factors which influence or predict its response are poorly understood.AimEvaluate the predictive factors for a successful long‐term therapeutic response of AZA.MethodsRetrospective analysis of all patients with IBD followed up in our hospital treated with AZA (2‐2.5mg/Kg/day) due to steroid dependent or resistent disease or, in Crohn disease (CD), due to fistulizing behavior or post‐surgery. We recorded the type of disease (DC/ ulcerative colitis (UC), indeterminate IBD), clinical parameters, laboratory parameters (LP) – WBC, CRP, hemoglobin, platelets and MCV – before and after 3 months of treatment, as well as concomitant usage of 5‐ASA and steroids. The treatment was considered effective when patients maintained control of the disease by clinical/endoscopic criteria, with continued maintainance of AZA or cessation of therapy after 3 months of treatment, and without escalation of therapy. We excluded patients who show intolerance to AZA in the first 3 months and patients treated concomitantly with biological agents.Results72 patients (37 women and 35 men); mean age 38.0±13.8 years; 35 patients with CD, 34 with UC and 3 with indeterminate IBD. The average duration of treatment with AZA was 35.1±30.6 months. AZA was effective in 48 patients (66.7%). The age at onset of AZA predicts therapeutic sucess (R=0.303, p=0.019). The sex, type of disease and LP before treatment did not correlate with efficacy. The LP after 3 months of therapy correlated with therapeutic sucess in the long‐term: WBC (r=–0.295, p=0.013), CRP (r=–0.332, p=0.005), hemoglobin (r=0.307, p=0.010), platelets (r=–0.360, p=0.003) and MCV (r=0.255, p=0.047). In combination, LP predict the efficacy of treatment (R=0.517, p=0.005). There is also an association between the location of UC (r=–0.381, p=0.026), as well as the duration of concurrent treatment with 5‐ASA (r=0.258, p=0.029) and the suspension of steroids (r=0.265, p=0.04) with the efficacy of the treatment.ConclusionAZA proved to be an effective treatment in the majority of patients with IBD. The old age of onset of the therapy and LP at 3 months were predictive of a sustained response of AZA

Characterization of Cynara cardunculus L. flower from Alentejo as a coagulant agent for cheesemaking

This work was supported by the project ValBioTecCynara (ALT2003-0145- FEDER-000038) - Economic valorisation of Cardoon (Cynara cardunculus): study of natural variability and biotechnological applications), cofinanced by FEDER under the Alentejo 2020 Program.The cardoon (Cynara cardunculus L.) is a mandatory vegetable coagulant for certain Protected Designation of Origin Portuguese cheeses. It grows wild in Portugal and is used without any type of control regarding flower picking or extract preparation, representing some uncertainty in cheese manufacture. The variability in technological properties, in the context of traditional cheese manufacture, of cardoon flower ecotypes from the Alentejo region was evaluated, including milk clotting and proteolytic activities, coagulation properties and potential cheesemaking yield of flower extracts. Multivariate statistics highlighted the variability of flower properties for cheesemaking, but allowed the aggregation of the ecotypes into five groups under the major influence of milk clotting activity and effect on gel firmness and micellar aggregation rate, followed by proteolytic activity. These differences may have an impact on cheese properties and therefore can allow the selection of cardoon flower for the manufacture of different types of cheese.publishersversionpublishe

Cynara cardunculus: Use in Cheesemaking and Pharmaceutical Applications

Cynara cardunculus L. is the most widespread species of Cynara genus (f. Asteraceae). This herbaceous perennial plant is native to the Mediterranean region and invasive in other parts of the world, growing naturally in harsh habitat conditions. There are three subspecies: globe artichoke; cultivated cardoon and the progenitor of the two, the wild cardoon. The culture of Cynara cardunculus L. follows an annual growth cycle, emerging in autumn and harvesting in summer. Cynara cardunculus has been considered as a multi-purpose crop due to its relevant biochemical profiles. Inflorescences have been used as food, whereas leaves are a rich source of bioactive compounds. Consequently, larger plants without spines have been selected for technological purposes. Due to its high cellulose and hemicellulose content, the lignocellulosic fraction has been used as solid biofuel, biogas and bioethanol. Both pulp fibers production and seeds oil are suitable for biodiesel production. Over the centuries, the inflorescence pistils of Cynara cardunculus L. have been widely used for cheesemaking. The present chapter gives an overview of the Cynara cardunculus L. emphasizing recent knowledge regarding the use, conservation, preparation and application of Cynara cardunculus in ovine milk cheesemaking, as well as other biotechnological applications

Mediterranean Shrubs: Nutraceuticals for sustainable goat production

Mediterranean Shrubs: Nutraceuticals for sustainable goat production Costa, C.ab, Almeida, S.M.c, Cavaco-Gonçalves, S.a, Barbas, J.P.a, Ribeiro, J.M.B.F.a, Belo, C.C.a, Padre, L.b, Belo, A.T,a aINIAV, Santarém – National Institute of Agrarian and Veterinarian Research, Quinta da Fonte Boa, 2005-048 Vale de Santarém bICAAM – Department of Veterinary Medicine, Évora University, Pólo da Mitra, 7006-554 Évora cUTAD – Trás-os Montes and Alto Douro University, Quinta de Prados, 5000-801 Vila Real E-mail: [email protected] Bioactive natural products have become an important research topic and development area in different fields of study, owing to their numerous potentialities. In animal production, the anthelminthic, antioxidant and antimicrobial properties of natural vegetation can improve the animals’ productive efficiency and wellbeing, and product quality (meat and milk). It is known that goats have a very selective feeding behavior and a higher sensitivity to gastrointestinal strongyles (GIS) infections compared to other ruminants, and that certain plants have beneficial effects in animal health through the synergy between their nutritive value and bioactive compounds. Thus, the project “VegMedCabras” intends to characterize and explore the nutraceutical value of mediterranean shrubs for sustainable goat production, by controlling parasitic infection as well as preventing environmental contamination and residues in the final product from the continuous use of synthetic anthelmintics [1,2,3]. Charnequeira breed goats were observed during browsing/grazing of Mediterranean vegetation, from February to July, in order to document their shrub species preferences and evaluate the nutraceutical value of the final diets. All selected plants were analyzed for nutritive value (protein and fiber content, digestibility), as well as bioactive compounds content such as total phenolic compounds (TPC), total tannins (TT), condensed tannins, flavonoids and saponins. Antioxidant activity was also evaluated. Each goat was individually evaluated for natural GIS infection, by egg elimination levels, and health status, by metabolic blood parameters. A significant drop in EPG (eggs per gram of feces) elimination (± 65%) was observed by the end of the study period. As soon as goats started browsing the Mediterranean vegetation, EPG counts started to decrease as the proportion of shrubs rich in TPC increased in the diet. Species with the highest contribution were Pistacia lentiscus and Quercus coccifera, which presented an average TPC and TT of 163,6 and 134,2 mg GAE (gallic acid equivalent)/g MS vs. 138,4 and 130,2 mg GAE/g MS, respectively. Nutritive value of the diets selected by goats was estimated in order to determine the adequacy to their physiological requirements. With the increased resistance of GIS to synthetic anthelmintics and social demand for quality products, it is important to establish multidisciplinary options such as the use of nutraceutical shrubs, a safer, ecological and inexpensive solution. Keywords: nutraceuticals; Mediteranean shrubs; bioactive compounds; anthelmintics; goats References: [1]. Jasmim RS (2018) Int. J. Botany 14: 24-29. [2]. Saric T, Rogosic J, Zupan I, Beck R, Bosnic S, Sikic Z, Skobic D, Tkalcic S (2015) Small Rum. Res. 123: 179-182. [3]. Hoste H, Torres-Acosta JFJ, Sandoval-Castro CA, Muller-Harvey I, Sotiraki S, Louvandini H, Thamsborg SM, Terrill TH (2015) Vet. Parasitology 212: 5-17. Acknowledgements: VegMedCabras – Mediterranean shrubs: natural anthelmintics in the diet selected by grazing goats (ALT-03-0145-FEDER-000009) co-financed by ALENTEJO2020 program-European Regional Development Fund

Chronic respiratory diseases and quality of life in elderly nursing home residents

Funding The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was supported by Fundação para a Ciência e Tecnologia–GERIA Project PTDC/SAU-SAP/ 116563/2010.Few studies have assessed the quality of life (QOL) related to chronic respiratory diseases in the elderly. In the framework of the geriatric study on the health effects of air quality in elderly care centers (GERIA) study, a questionnaire was completed by elderly subjects from 53 selected nursing homes. It included various sections in order to assess respiratory complaints, QOL (World Health Organization QOL (WHOQOL)-BREF), and the cognitive and depression status. The outcome variables were the presence of a score lower than 50 (publishersversionpublishe