Caracterização química do grão de ervilha proteaginosa

Seminário realizado na Escola Superior Agrária do Instituto Politécnico de Castelo Branco, no âmbito do Projecto 0186_AGROCELE_3_EO objectivo deste trabalho foi determinar a matéria seca (MS), cinzas, matéria orgânica (MO), proteína bruta (PB), gordura bruta (GB), açúcares totais, amidos e estimar, a partir da composição química, a EM ruminantes, EM suínos, EM aves, ED coelhos e ED cavalos de cada uma de 10 cultivares mais produtivas obtidas num ensaio com 20 cultivares de ervilha proteaginosa (Pisum sativum) realizado na ESACB (Novembro de 2009 a Junho de 2010) (Projecto 0186_AGROCELE_3_E). As análises químicas foram realizadas no Laboratório de Nutrição e Alimentação Animal da ESACB. Para cada cultivar obtiveram-se 4 resultados que foram tratados estatisticamente (média, desvio padrão, ANOVA, teste Duncan). Os resultados obtidos (% na MS) permitem-nos afirmar o seguinte: as 10 variedades de ervilha proteaginosa estudadas constituem importante fonte de energia disponível (glúcidos citoplásmicos) com elevadas percentagens de açúcares solúveis (7,95% ISARD a 9,42% ENDURO) (P0,05), EM suínos (13,57 MJ/kg MS CORRENT a 14,71 MJ/kg MS ALHAMBRA) (P>0,05); EM aves (11,96 MJ/kg MS ENDURO a 12,45 MJ/kg MS AUDIT) (P>0,05), ED cavalos (13,73 MJ/kg MS CORRENT a 14,37 MJ/kg MS ALHAMBRA) (P>0,05) nas diferentes cultivares e apresentam ligeiras diferenças na ED coelho (12,99 MJ/kg MS CORRENT a 13,03 MJ/kg MS ALHAMBRA) (P<0,05). Conclui-se que a ervilha proteaginosa é um excelente alimento para ruminantes e monogástricos, podendo ser fornecida simples ou incluída em alimentos concentrados; é excelente como suplemento energético e proteico uma vez que associa, no mesmo grão, elevados níveis de PB e de amido; apresenta um valor energético relativamente elevado; devido ao baixo teor de GB é um alimento muito interessante para ruminantes e para dietas light de animais de companhia; dependendo do preço de mercado pode vir a substituir, total ou parcialmente, o milho (como fonte de glúcidos facilmente fermentescíveis) e a soja (como principal fonte de proteína).Instituto Tecnológico Agrário de Castilla y León y Instituto Politécnico de Castelo Branco/Escola Superior Agrári

Evaluation of proteaginous Pisum sativum L. cultivars in the region of Castelo Branco

Evaluation of proteaginous Pisum sativum L. cultivars in the region of Castelo Branco Carlos M. G. Reis(1)* and Paulo Rodrigues(1) 1 Escola Superior Agrária, Instituto Politécnico de Castelo Branco. *[email protected] The pea crop (Pisum sativum L.) is a convenient source of plant protein for animal feeding, an area where there is a production deficit in European Union. After obtaining new cultivars through plant breeding it is important to evaluate their agronomic performance in different regions. This study aimed to evaluate the agronomic performance of 20 cultivars of proteaginous Pisum sativum L., listed in the Community Catalogue of varieties of agricultural plant species, in the region of Castelo Branco, Portugal. A field trial was implanted in Escola Superior Agrária de Castelo Branco. Sowing took place on November 2009 in plots with 12.0 m2 in a randomized complete block design with four replications. A density of 110 plants per m2 was used. Some parameters related to plant growth and yield were studied, such as seed yield (kg/ha), seed moisture content (%), weight of 1000 seeds (g), number of days to flowering, number of days to harvest, lodged plants (%), dehiscence (%), plant height (cm), dry matter weight, biological weight, number of plants per m2, number of seeds per m2, height of first pod (cm), number of pods per plant, number of seeds per pod and seed number per plant. The seed protein content was also studied but only for the 10 highest yielding cultivars. The statistical analysis was performed using PASW Statistics 18 software. Analysis of variance (ANOVA) for significance level p=0.05 and the mean comparison by Duncan test application were conducted. For some yield components we calculated the Pearson correlation coefficient. The cultivars studied showed significant differences in all quantitative traits studied. With regard seed yield, there were values greater than 6,000 kg/ha for 10 cultivars (Cartouche, Enduro, Arthur, Audit, Corrent, Alhambra, Cherokee, Isard, Livia and Gregor) and 16 cultivars showed productions above 4,000 kg/ha. However, these results cannot be dissociated from the precipitation values recorded, well above the normal for the region. Among the best cultivars, Enduro and Cartouche are those with the lowest percentage of lodged plants. The cultivars Arthur, Corrent, Cherokee, Livia, Pixel, Ideal, Guifilo, Guifredo, Lumina and Grisel, showed a strong tendency to lodging. In general there is a positive correlation between seed yield and other quantitative variables, except the weight of 1000 seeds. The positive correlations were highest to the number of seeds per m2 (0.847), biological weight (0.787) and harvest index (0.857). The seed protein content was similar for the cultivars studied. Although the results of seed yield are very interesting, it is necessary to conduct additional trials to evaluate the agronomic performance of pea cultivars in order to obtain more consistent results. However, the results allow us to elect a group of cultivars with high seed yield and apparently good adaptability to the region of Castelo Branco

Modelling the complex nature of the tumor microenvironment: 3D tumor spheroids as an evolving tool

Cancer remains a serious burden in society and while the pace in the development of novel and more effective therapeutics is increasing, testing platforms that faithfully mimic the tumor microenvironment are lacking. With a clear shift from animal models to more complex in vitro 3D systems, spheroids emerge as strong options in this regard. Years of development have allowed spheroid-based models to better reproduce the biomechanical cues that are observed in the tumor-associated extracellular matrix (ECM) and cellular interactions that occur in both a cellâ cell and cell-ECM manner. Here, we summarize some of the key cellular interactions that drive tumor development, progression and invasion, and how successfully are these interactions recapitulated in 3D spheroid models currently in use in the field. We finish by speculating on future advancements in the field and on how these can shape the relevance of spherical 3D models for tumor modelling.Authors would like to acknowledge the fnancial support from the European Research Council through the Starting Grant “CapBed” (ERC-2018-STG-805411) and FCT/MCTES (Fundação para a Ciência e a Tecnologia/ Ministério da Ciência, Tecnologia, e Ensino Superior) through the grant SFRH/BD/119756/2016 (D.B.R.). The authors would additionally like to thank the contributions to this work from the project “TERM RES Hub—Scientifc Infrastructure for Tissue Engineering and Regenerative Medicine”, reference PINFRA/22190/2016 (Norte-01-0145-FEDER-022190), funded by the Portuguese National Science Foundation (FCT) in cooperation with the Northern Portugal Regional Coordination and Development Commission (CCDR-N). Ultimately, we would like to equally acknowledge fnancial support fromhttps://doi.org/10.54499/ UIDB/50026/2020);https://doi.org/10.54499/UIDP/50026/2020) andhttps://doi. org/10.54499/LA/P/0050/2020)

Adult Pancreatoblastoma - Case Report and Review of Literature

Most cases of pancreatoblastoma, a rare malignant epithelial tumor of the pancreas, are seen in the pediatric population. The rarity of pancreatoblastoma, the similar radiologic findings to those seen in other pancreatic lesions, and its histopathologic heterogeneity, make its preoperative diagnosis in adults a real challenge. We report ultrasound, computed tomography and magnetic resonance imaging correlative findings of a histologically proven pancreatoblastoma in a 37-year-old woman. Pancreatoblastoma should be considered in the differential diagnosis of a pancreatic mass presenting uncommon imaging features.info:eu-repo/semantics/publishedVersio

Unravelling the path to create a cell sheet-based model of skin scar-like tissue

Regardless of the advances in understanding the mechanisms and the pathophysiology behind skin deformities, scaring continues to be an unsolved clinical problem. The underlying wound healing process involves a series of key cells which play different key roles. Fibroblasts are known to suffer the influence of local biochemical (e.g TGF-B1) and biomechanical signaling upon a wound scenario leading to a phenotypical change into myofibroblasts. The latter enhance immature extracellular matrix (ECM) synthesis and generate tensional forces that leads to ECM reorganization. Certain skin pathologies (e.g hypertrophic scars) rise from a dysfunction of this underlying regulatory mechanism which in turn drives myofibroblast persistence in the wound. When trying to study the mechanisms behind scarring human ex vivo samples are many times scarce and most of the current in vitro systems rely on standard 2D cultures of keloid/hypertrophic scar fibroblasts. Taking all of this into consideration we propose the use of cell sheet technology to create an in vitro 3D scar model. Herein we report the effect of TGF-B1 in human dermal fibroblast cell sheets as the first step to attain cell sheets with a myofibroblast-like phenotype in which cells are embedded in a scar-like ECM. To further strengthen our concept we performed the stacking of pre-formed cell sheets generating a cohesive 3D scar-like tissue. Human dermal fibroblast (hDFbs) cell sheets were produced as previously described1, and stimulated with TGF-B1 (10ng/ml) over 7, 14 and 21 days. Following phenotype and ECM characterization, cell sheets were stacked in order to obtain a 3D structure composed of 2 or 3 cell-sheets. The analysis of key genes (q-PCR) and proteins (Western blot and immunocytochemistry) showed that hDFbs cell sheets, when stimulated with TGF-B1 present an increased expression of a-SMA, fibronectin (FN) ED- A and FN ED-B, characteristic of a myofibroblast-like phenotype. When looking into the expression of scar ECM-associated proteins, hDFbs cell sheets obtained in the presence of TGF-B1 produced higher amounts of fibronectin and collagen I. Stable 3D constructs with a noticeable level of integration after a total of 21 days of culture, were further created upon stacking of the cell sheets obtained after 7days of culture in the presence of TGF-B1. In conclusion, this work suggested that it is possible to promote the secretion of scar-like ECM in hDFbs cell sheets due to phenotypic changes into myofibroblast-like cells when stimulated with TGF-B1. Cohesive 3D scar-like tissue structures were obtained which opens the possibility to develop a highly accurate in vitro 3D scar model to study underlying cellular mechanisms involved in the wound healing deregulation. Grant IF/00945/2014 funded by FCT/MCTES, Project “NORTE-08-5369-FSE-000044”, funded by Programa Operacional Norte 2020 Fundo Social Europeu, and GENE2SKIN Twinning Project, Horizon 2020, funded by the European Commissioninfo:eu-repo/semantics/publishedVersio

In vitro 3D cell sheet-based model for unraveling scar pathophysiology

Fibroblasts are key players in the scarring process. In hypertrophic scars, fibroblasts suffer phenotypical changes into myofibroblasts persisting in the wound under the influence of local biochemical (TGFb1) and biomechanical signaling leading to enhanced immature extracellular matrix (ECM) synthesis. Benchtop models of hypertrophic scars rely on scarce human ex vivo samples or standard 2D cultures of hypertrophic scar fibroblasts. We therefore propose the use of human dermal fibroblast cell sheets (hDFbsCS) as the first step to attain cell sheets with a myofibroblast-like phenotype to generate cohesive in vitro 3D scar-like tissues. hDFbsCS were produced as previously described (Cerqueira, 2014), and stimulated with TGFb1 up to 21 days. Following phenotype and ECM characterization, 3 hDFbsCS were stacked to obtain a 3D structure. Gene and protein analysis showed that upon TGFb1 stimulation, hDFbsCS present a higher expression of aSMA, fibronectin EDA and EDB, characteristic of amyofibroblast-like phenotype. Regarding the expression of scar ECM-associated proteins, TGFb1 stimulated hDFbsCS produced increased fibronectin and collagen I. Upon stacking of hDFbsCS obtained after 7 days of culture in the presence of TGFb1, stable and integrated 3D constructs were obtained. This work suggests that it is possible to create cohesive 3D scar-like tissue structures from hDFbsCS opening the possibility to develop in vitro 3D scar models to study wound healing deregulation pathophysiology. Acknowledgments: Grant IF.00945.2014 and SFRH.BD. 119756.2016 (FCT MCTES), NORTE.08.5369.FSE.000044 (funded by Programa Operacional Norte 2020 Fundo Social Europeu), GENE2SKIN Twinning Project, Horizon 2020 (European Commission).Grant IF.00945.2014 and SFRH.BD.119756.2016 (FCT_MCTES), NORTE.08.5369.FSE.000044 (funded by Programa_Operacional_Norte_2020 Fundo Social Europeu), GENE2SKIN Twinning Project, Horizon_2020 (European Commission).info:eu-repo/semantics/publishedVersio

Chitosan/virgin coconut oil-based emulsions doped with photosensitive curcumin loaded capsules: a functional carrier to topical treatment

In recent years, there has been a growing interest in developing smart drug delivery systems based on natural resources combined with stimulus-sensitive elements. This trend aims to formulate innovative and sustainable delivery platforms tailored for topical applications. This work proposed the use of layer-by-layer (LbL) methodology to fabricate biocompatible photo-responsive multilayer systems. These systems are composed of a polyoxometalate inorganic salt (POM) ([NaP5W30O110]14â) and a natural origin polymer, chitosan (CHT). Curcumin (CUR), a natural bioactive compound, was incorporated to enhance the functionality of these systems during the formation of hollow capsules. The capsules produced, with sizes between 2â 5μm (SEM), were further dispersed into CHT/VCO (virgin coconut oil) emulsion solutions that were casted into molds and dried at 37 â ¦C for 48 h.The system presented a higher water uptake in PBS than in acidic conditions, still significantly lower than that earlier reported to other CHT/VCO-based systems. The drug release profile is not significantly influenced by the medium pH reaching a maximum of 37% ± 1% after 48 h. The antioxidant performance of the designed structures was further studied, suggesting a synergistic beneficial effect resulting from CUR, POM, and VCO individual bioactivities. The increased amount of those excipients released to the media over time promoted an increase in the antioxidant activity of the system, reaching a maximum of 38.1% ± 0.1% after 48 h. This work represents a promising step towards developing advanced, sustainable drug delivery systems for topical applications.The authors would like to thank the contributions to this research from the project “TERM RES Hub–Scientific Infrastructure for Tissue Engineering and Regenerative Medicine”, reference PINFRA/22190/2016 (Norte-01-0145-FEDER-022190), funded by the Portuguese National Science Foundation (FCT) in cooperation with the Northern Portugal Regional Coordination and Development Commission (CCDR-N), for providing relevant lab facilities, state-of-the art equipment and highly qualified human resources

Stimulatory effects of inorganic ions on osteogenesis in vitro

Introduction: Several studies demonstrated the effect of silicate ions (Si) on differentiation of bone precursor cells1,2, although its exact role in processes related to bone formation and remodeling is still incompletely understood. The focus of this work is to explore the effect of calcium and silicate ions on growth and osteogenic differentiation of human mesenchymal stem cells (hMSCs). This strategy may reduce the need for growth factors required to stimulate bone formation in regenerative approaches, decreasing the associated costs and overcoming stability issues. Materials and Methods In order to define the range of Si concentrations that are not toxic to cells, we performed a preliminary study varying Si concentrations from 0.00357mM to 4mM. The concentration of the Ca ions was selected based on the earlier study by Barradas et. al3. Cell culture media were supplemented by using sodium silicate (Na2SiO3) and/or calcium chloride dehydrate (CaCl2*2H2O) as Si and Ca precursors, respectively. hMSCs derived from bone marrow were seeded at a seeding density of 2.000 cells/cm2 and allowed to adhere overnight. Then, the medium was replaced by the appropriate supplemented medium and cells were cultured for 3, 7, 14 and 18 days. Basic and osteogenic media were used as negative and positive controls. Cell proliferation was evaluated by DNA quantification. hMSCs osteogenic gene expression was evaluated by Q-PCR. Results DNA quantification indicated an increase in cell number during the culture time for all the conditions. Results obtained by Q-PCR revealed a significantly higher expression of osteocalcin (OC) and bone morphogenetic protein-2 (BMP2) in cells cultured in media supplemented by both ions, as compared to media containing either Ca or Si alone. Discussion and Conclusions DNA quantification studies indicated that none of the selected concentrations had a negative influence on cell proliferation. The increase in osteogenic gene expression for cells cultured with both Ca and Si suggested a synergistic effect of the two ions on osteogenic differentiation of hMSCs. We further showed that cells cultured in the medium with the highest concentration of Ca (7.8mM) revealed a higher expression of the selected genes, which is in accordance with the earlier results by Barradas et al3. The obtained results suggest the importance of combining both ions, Ca and Si, for promoting the osteogenic differentiation of hMSCs. References 1. Hoppe A, Biomaterials 32: 2757-2774, 2011. 2. Beck Jr GR, Nanomedicine: Nanotechnology, Biology, and Medicine,1-11, 2011 3. Barradas AMC et al., Biomaterials 3205-3215, 2012. Acknowledgments The author thanks the Portuguese Foundation for Science and Technology (FCT) for the grant (SFRH/BD/69962/2010). Disclosures The authors have nothing to disclose