Perfil toxicológico e efeito antioxidante e antiglicante in vitro de um extrato glicólico do resíduo do fruto da Punica granatum: Toxicological profile and in vitro antioxidant and antiglycolic effect of a glycolic extract of the fruit residue of Punica granatum

A casca do fruto da romãzeira (Punica granatum), a romã, é um resíduo produzido em abundância em território nacional. Sabendo-se que a espécie é rica em compostos fenólicos, metabólitos secundários com relevantes atividades terapêuticas, entre elas a antioxidante e antiglicante, e buscando suprir a demanda de fitocosméticos eficazes e seguros que possam prevenir o envelhecimento da pele; o presente estudo teve como objetivo analisar os aspectos fitoquímicos, antioxidantes e antiglicantes de um extrato produzido a partir da casca da romã, visando o aproveitamento integral do fruto. Para tanto, realizou-se os seguintes estudos in vitro: 1) fitoquímicos (dosagem de flavonoides, taninos condensados e hidrolisáveis); 2) citotoxicidade (MTT); 3) farmacológicos - atividade antioxidante (DPPH e lipoperoxidação) e atividade antiglicante no ensaio de albumina sérica bovina (BSA) com glicose e metilglioxal (MGO). O extrato apresentou o seguinte perfil fitoquímico de compostos fenólicos: flavonoides 1.9 ± 0.3 g de equivalente em quercetina para cada 100 g de extrato seco; taninos condensados 40.36 ± 9.5 g de equivalente em proantocianidina para cada 100 g de extrato seco e taninos hidrolisáveis 20.22 ± 6.3 de equivalente em ácido gálico para cada 100 g de extrato seco. Além de que no ensaio de MTT, a Concentração Citotóxica 50% para 24 h foi 899 ug/mL, para 48 h foi de 482 ug/mL e para 72 h foi 426 ug/mL. A atividade farmacológica, apresentou na concentração de 600 µg/mL uma atividade antioxidante para DPPH de 78.3% e uma atividade antiglicante de 62.19% no ensaio de lipoperoxidação; no sistema BSA/Glicose todas as concentrações promoveram 100% de atividade antiglicante e no BSA/MGO a concentração de 600 µg/mL promoveu atividade antiglicante em 40%. O extrato, portanto, se demonstra como um potencial candidato para formulação de um novo fitocosmético que possa ser utilizado na prevenção do envelhecimento da pele

Basin-wide variation in tree hydraulic safety margins predicts the carbon balance of Amazon forests

Funding: Data collection was largely funded by the UK Natural Environment Research Council (NERC) project TREMOR (NE/N004655/1) to D.G., E.G. and O.P., with further funds from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES, finance code 001) to J.V.T. and a University of Leeds Climate Research Bursary Fund to J.V.T. D.G., E.G. and O.P. acknowledge further support from a NERC-funded consortium award (ARBOLES, NE/S011811/1). This paper is an outcome of J.V.T.’s doctoral thesis, which was sponsored by CAPES (GDE 99999.001293/2015-00). J.V.T. was previously supported by the NERC-funded ARBOLES project (NE/S011811/1) and is supported at present by the Swedish Research Council Vetenskapsrådet (grant no. 2019-03758 to R.M.). E.G., O.P. and D.G. acknowledge support from NERC-funded BIORED grant (NE/N012542/1). O.P. acknowledges support from an ERC Advanced Grant and a Royal Society Wolfson Research Merit Award. R.S.O. was supported by a CNPq productivity scholarship, the São Paulo Research Foundation (FAPESP-Microsoft 11/52072-0) and the US Department of Energy, project GoAmazon (FAPESP 2013/50531-2). M.M. acknowledges support from MINECO FUN2FUN (CGL2013-46808-R) and DRESS (CGL2017-89149-C2-1-R). C.S.-M., F.B.V. and P.R.L.B. were financed by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES, finance code 001). C.S.-M. received a scholarship from the Brazilian National Council for Scientific and Technological Development (CNPq 140353/2017-8) and CAPES (science without borders 88881.135316/2016-01). Y.M. acknowledges the Gordon and Betty Moore Foundation and ERC Advanced Investigator Grant (GEM-TRAITS, 321131) for supporting the Global Ecosystems Monitoring (GEM) network (gem.tropicalforests.ox.ac.uk), within which some of the field sites (KEN, TAM and ALP) are nested. The authors thank Brazil–USA Collaborative Research GoAmazon DOE-FAPESP-FAPEAM (FAPESP 2013/50533-5 to L.A.) and National Science Foundation (award DEB-1753973 to L. Alves). They thank Serrapilheira Serra-1709-18983 (to M.H.) and CNPq-PELD/POPA-441443/2016-8 (to L.G.) (P.I. Albertina Lima). They thank all the colleagues and grants mentioned elsewhere [8,36] that established, identified and measured the Amazon forest plots in the RAINFOR network analysed here. The authors particularly thank J. Lyod, S. Almeida, F. Brown, B. Vicenti, N. Silva and L. Alves. This work is an outcome approved Research Project no. 19 from ForestPlots.net, a collaborative initiative developed at the University of Leeds that unites researchers and the monitoring of their permanent plots from the world’s tropical forests [61]. The authros thank A. Levesley, K. Melgaço Ladvocat and G. Pickavance for ForestPlots.net management. They thank Y. Wang and J. Baker, respectively, for their help with the map and with the climatic data. The authors acknowledge the invaluable help of M. Brum for kindly providing the comparison of vulnerability curves based on PAD and on PLC shown in this manuscript. They thank J. Martinez-Vilalta for his comments on an early version of this manuscript. The authors also thank V. Hilares and the Asociación para la Investigación y Desarrollo Integral (AIDER, Puerto Maldonado, Peru); V. Saldaña and Instituto de Investigaciones de la Amazonía Peruana (IIAP) for local field campaign support in Peru; E. Chavez and Noel Kempff Natural History Museum for local field campaign support in Bolivia; ICMBio, INPA/NAPPA/LBA COOMFLONA (Cooperativa mista da Flona Tapajós) and T. I. Bragança-Marituba for the research support.Tropical forests face increasing climate risk1,2, yet our ability to predict their response to climate change is limited by poor understanding of their resistance to water stress. Although xylem embolism resistance thresholds (for example, Ψ50) and hydraulic safety margins (for example, HSM50) are important predictors of drought-induced mortality risk3-5, little is known about how these vary across Earth's largest tropical forest. Here, we present a pan-Amazon, fully standardized hydraulic traits dataset and use it to assess regional variation in drought sensitivity and hydraulic trait ability to predict species distributions and long-term forest biomass accumulation. Parameters Ψ50 and HSM50 vary markedly across the Amazon and are related to average long-term rainfall characteristics. Both Ψ50 and HSM50 influence the biogeographical distribution of Amazon tree species. However, HSM50 was the only significant predictor of observed decadal-scale changes in forest biomass. Old-growth forests with wide HSM50 are gaining more biomass than are low HSM50 forests. We propose that this may be associated with a growth-mortality trade-off whereby trees in forests consisting of fast-growing species take greater hydraulic risks and face greater mortality risk. Moreover, in regions of more pronounced climatic change, we find evidence that forests are losing biomass, suggesting that species in these regions may be operating beyond their hydraulic limits. Continued climate change is likely to further reduce HSM50 in the Amazon6,7, with strong implications for the Amazon carbon sink.Publisher PDFPeer reviewe

Práticas Educomunicativas

Esta publicação pretende divulgar as práticas educomunicativas realizadas em diferentes regiões do país e que estão sendo implantadas por nossos associados. O e-book Práticas Educomunicativas, que visa oferecer um material de uso prático que possa servir de apoio pedagógico em diferentes contextos, escolar ou de ações junto a instituições, apresenta 20 artigos de profissionais e pesquisadores que implementam ações que inter-relacionam comunicação e educação no contexto da educação apontando as experiências e processos de educomunicação e valorizando desta forma, o trabalho realizado por cada educomunicador oferecendo, ao leitor, um material de uso prático que possa servir de apoio pedagógico em diferentes contextos

Cultura de condrócitos para o uso terapêutico: reconstituição de cartilagem

A cartilagem articular é um tecido avascular com números limitados de condrócitos, com capacidade limitada de reparo após uma lesão aguda. As técnicas disponíveis atualmente para o tratamento de lesões de cartilagem articular podem resultar em alívio dos sintomas, mas não na regeneração do tecido lesado. A geração de um substituto biológico que recomponha a cartilagem articular nativa requer células vivas que sejam capazes de sintetizar e manter a matriz cartilaginosa. A engenharia de tecidos constitui recentemente uma metodologia para reconstrução de novos órgãos e tecidos que foram lesados e apresentam dificuldades na reparação. Um dos maiores avanços no campo da engenharia de tecidos e dentro da medicina ortopédica, tem sido, o recrutamento de tecido do próprio paciente, que são dissociados em células e cultivadas sobre suportes biológicos ou sintéticos conhecidos como scaffolds, para posterior realização de implante de condrócitos, com intuito de regenerar o tecido cartilaginoso lesado. Uma variedade de scaffolds como hidrogel e polímeros sintéticos, têm sido investigadas para a expansão dos condrócitos “in vitro” para o reparo da cartilagem lesada. Tais matrizes incluem: arcabouços à base de colágeno: gel de colágeno tipo I e II, esponjas de colágeno tipo II, ácido polilático e ácido poliglicólico, fibrina, óxido de polietileno, fibrina, peptídeos, alginato e gel de plaquetas. No presente trabalho desenvolvemos diferentes metodologias de cultura de condrócitos utilizando scaffold sintético “hidrogel de alginato” e biológico “gel de plaquetas” obtido a partir do Plasma Rico em Plaquetas - PRP, além da cultura de condrócitos em pellet tri-dimensional, objetivando encontrar a melhor metodologia para a realização de implantes de condrócitos. Como modelo experimental escolhemos coelhos, tanto para coleta da cartilagem articular...Articular cartilage is an avascular tissue with limited numbers of chondrocytes, and limited recuperative capacity after a severe lesion. The techniques currently available for treatment of articular cartilage lesions can result in relief of symptoms, but not in regeneration of injured tissue. The generation of biological substitute that would recompose native articular cartilage requires live cells capable of synthesizing and maintaining cartilaginous matrix. The engineering of tissues has recently constituted a methodology for constructing new organs and tissues that had been injured and present recuperative difficulties. One of the greatest advances in the field of tissue engineering, and within orthopedic medicine, has been the recruitment of tissue from the patient him- or herself, that are dissociated in cells and cultured on biological or synthetic supports, known as scaffolds, for subsequent realization of implantation of chondrocytes, in order to regenerate injured cartilaginous tissue. A variety of scaffolds, such as hydrogel and synthetic polymers, have been investigated for expansion of chondrocytes in vitro to repair injured cartilage. Such matrices include: collagen-based scaffolds: gel of collagen types I and II, sponges of collagen type II, polylactic acid and polyglycolic acid, fibrin, polyethylene oxide, peptides, alginate and gel of platelets. In the present work we developed different methodologies of chondrocyte culture utilizing the synthetic scaffold “alginate hydrogel” and biological “gel of platelets” obtained from Platelet-Rich Plasma - PRP, in addition to chondrocyte culture in a tri-dimensional pellet, aiming to find the best methodology for accomplishing chondrocyte implants. We chose rabbits for the experimental model, as much for collection of articular cartilage as for obtaining blood plasma and realization of allogenic implants... (Complete abstract click electronic access below)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Cultura de condrócitos em arcabouço tridimensional: hidrogel de alginato Chondrocyte cultures in tridimensional scaffold: alginate hydrogel

OBJETIVOS: O presente estudo teve como objetivo cultivar condrócitos retirados da articulação do joelho de coelhos encapsulados em hidrogel de alginato (HA) e caracterizar a produção de matriz extracelular (ECM). MÉTODOS: A cartilagem articular foi removida do joelho de coelhos, com três a seis meses, fragmentada em pedaços de 1mm e submetida à digestão enzimática. Uma concentração de 1x106 céls/mL foram ressuspensas em uma solução de alginato de sódio a 1,5% (w/v), em seguida fez-se o processo de gelatinização em CaCl2 (102 mM), permitindo a formação do HA e cultivo em meio DMEM-F12 durante quatro semanas. A distribuição das células e a ECM foram acessadas através das secções histológicas coradas com e azul de toluidina hematoxilina e eosina (HE). RESULTADOS: Houve um aumento no número e na viabilidade dos condrócitos durante as quatro semanas de cultura. Através das análises histológicas dos HAs corados com azul de toluidina e HE foi possível observar a distribuição definida dos condrócitos no hidrogel, assemelhando-se a grupos isógenos e formação de matriz territorial. CONCLUSÃO: Este estudo demonstrou a eficiência do HA como arcabouço para ser usado na cultura de condrócitos, constituindo uma alternativa no reparo de lesões na cartilagem articular.<br>OBJECTIVES: The aim of this study was to culture chondrocytes from knee joint cartilage of rabbits encapsulated in alginate hydrogel (HA) and to characterize the production of extracelular matrix (ECM). METHODS: Joint cartilage was obtained from rabbits' knees, three to six months old, fragmented into 1-mm pieces and submitted to enzymatic digestion. A concentration of 1x106 cells/mL were re-suspended into a 1.5% (w/v) sodium alginate solution, followed by gel formation process with CaCl2 (102 mM), allowing HA to build for culturing it into a DMEM-F12 medium for four weeks. The distribution of cells and ECM were assessed from histological slices stained toluidine blue and hematoxyline-eosin (HE). RESULTS: There was an increase of the number and viability of the chondrocytes during the four weeks of culture. By assessing the histological sections stained with toluidine blue and HE, we could note the definitive distribution of chondrocytes in the hydrogel, similarly to isogenous groups and territorial matrix formation. CONCLUSION: In this study, the alginate was shown to be an effective scaffold for use in chondrocytes culture, constituting an alternative for repairing joint cartilage defects

Toxicity and Anti-Inflammatory Effects of <i>Agave sisalana</i> Extract Derived from Agroindustrial Residue

Background: In several countries, the leaf juice of Agave sisalana (also known as sisal) is widely used topically, especially as an antiseptic, and orally for the treatment of different pathologies. However, in Brazil, which is the largest producer of Agave sisalana, its residue, which represents the majority of its weight, has been thrown away. For this reason, the determination of the pharmacological and toxicological potentials of sisal residue and its possible therapeutic use is seen as a way to contribute to the sustainable development and social promotion of the largest producer of sisal in Brazil, the interior of Bahia State, which is among the poorest areas in the country. Given the scarcity of available scientific studies on the pharmacological and toxicological properties of sisal residue juice, this study aimed to promote the acid hydrolysis of this juice to potentiate the anti-inflammatory effect already described in the literature. Furthermore, it aimed to evaluate the toxicological profile of the hydrolyzed extract (EAH) and to determine its acute toxicity, as well as its side effects on the reproductive aspects of rats. Method: The anti-inflammatory effect of EAH was evaluated in vitro using the induction of hemolysis by hypotonic solution and in vivo in rats using the carrageenan-induced paw edema test and the xylene-induced ear edema test. The acute toxicity, resulting from a single-dose administration, was investigated for some manifestation of toxic symptoms related to motor control and consciousness in rats. At a concentration of 100 mg/kg, by repeated doses, the reproductive toxicity effects of EAH in rats were assessed. Results: In vitro anti-inflammatory activity was positive using the human red blood cell membrane stabilization method. In both in vivo tests used to assess the anti-inflammatory activity, EAH (at three doses) significantly inhibited edema when compared to the control group. At a dose of 50 mg/kg, EAH exhibited a greater effect than indomethacin, a nonsteroidal anti-inflammatory drug with known activity. In vivo toxicological studies have shown that EAH does not present toxic effects when administered orally in a single dose, up to 1000 mg/kg. Finally, EAH promoted a gonadotoxic effect and increased the embryonic mortality rate after implantation. Conclusions: It is suggested that the anti-edematogenic effect of the acid hydrolysis extract from sisal juice is due to the high concentration of steroidal sapogenins. Therefore, this extract can be considered a potential new anti-inflammatory or even an important sapogenin source for the development of steroidal glucocorticoids. However, further studies are needed to elucidate the chemical composition of sisal juice. Regarding toxicology studies, EAH did not show cytotoxic and clastogenic potentials, but it presented a powerful reproductive toxic effect in rats

Terapia celular em modelo experimental de doença inflamatória intestinal

Inflammatory bowel disease, which mainly involves Crohn's disease and ulcerative rectocolitis, is an inflammatory condition of the mucosa that can afflict any segment of the gastrointestinal tract. Despite the fact that the existing therapies result in improvement in patient's symptomatology and quality of life, there is no curative treatment. Surgical treatment involves complex procedures associated with high morbidity and mortality rates. In this context, cell therapy with stem cells has emerged as a treatment with broad potential applicability. In this study, we intended to verify the efficacy of transplantation of adipose tissue-derived stem cells in rats with intestinal inflammation induced by trinitrobenzenesulfonic acid. The cell population was isolated from the adipose tissue of inguinal region of rats and processed for culture by mechanical dissociation. The animals were evaluated with respect to clinical and biochemical aspects, as well as by macroscopic, microscopic and histological analyses. In the experimental model of bowel inflammation by 2,4,6-trinitrobenzenesulfonic acid, the infusion of adipose tissue significantly reduced the presence of adhesions in the colon and adjacent organs and decreased the activity of myeloperoxidase, a marker of neutrophil infiltration in the injured mucosa. The results suggest that cell therapy with adipose tissue can promote and/or accelerate the regeneration of damaged intestinal mucosa. It is concluded that the presence of adhesions and the determination of myeloperoxidase activity provide indications that adipose tissue can promote and/or accelerate the regeneration of inflammatory bowel mucosa.A Doença Inflamatória Intestinal (DII), consistindo principalmente da doença de Crohn e retocolite ulcerativa, é uma condição inflamatória da mucosa que pode acometer qualquer segmento do trato gastrointestinal. Apesar das terapias existentes resultarem na melhora dos sintomas e da qualidade de vida dos pacientes, não há nenhum tratamento curativo. O tratamento cirúrgico envolve procedimentos complexos associados a altas taxas de morbimortalidade. Neste contexto, a terapia celular com células-tronco desponta como opção de tratamento potencialmente promissora. Em função destes aspectos, pretendeu-se, no presente estudo, verificar a eficácia do transplante de células-tronco derivadas do tecido adiposo (ASC) em ratos com inflamação intestinal induzida por ácido trinitrobenzenosulfonico (TNBS). As ASCs foram obtidas por dissociação mecânica do tecido adiposo da região inguinal de ratos e processadas para cultivo. Os animais foram avaliados, considerando-se os aspectos clínicos e bioquímicos, além de análises macroscópica, microscópica e histológica. No modelo de inflamação intestinal induzida por TNBS, a infusão de ASCs reduziu significativamente a presença de aderências entre o cólon e órgãos adjacentes, bem como diminuiu a atividade da mieloperoxidase (MPO), um marcador da infiltração de neutrófilos na mucosa lesada. Os resultados obtidos permitem concluir que a terapia celular com ASCs pode promover e/ou acelerar o processo de regeneração da mucosa intestinal inflamada

Isolamento de células-tronco mesenquimais da medula óssea Isolation of bone marrow mesenchymal stem cells

As Células-Tronco Mesenquimais (CTMs) têm alta capacidade de se renovar e diferenciar em várias linhagens de tecido conjuntivo. Este trabalho teve como objetivo isolar as CTMs da medula óssea de camundongos utilizando dois diferentes meios de cultura e caracterizá-las através de imuno-marcação com anti-vimentina. Foram utilizados 6 camundongos BALB/c com 15 dias de idade. A medula óssea foi coletada do canal medular das tíbias e fêmures dos camundongos e ressuspensas em uma concentração final 6x10(5), em meio Knockout- DMEM e DMEM alta concentração de glicose, suplementados com 10% SBF, mantidas em estufa a 37° C em uma atmosfera úmida a 5% de CO2 e 95% de ar por 72 horas, quando as células não aderentes foram removidas durante a troca do meio. O número e densidade de células com morfologia fibroblastóide foram maior no meio Knockout- DMEM em cinco dias de cultura versus 10-20 dias para conseguir a mesma concentração celular com o DMEM alta concentração de glicose. As células de ambos grupos apresentaram intensa marcação com anticorpo anti-vimentina, caracterizando-as como CTMs. A obtenção mais rápida das CTMs é fundamental para o campo da terapia celular, principalmente quando se deseja utilizar estas células no reparo de tecidos de origem mesenquimal.<br>Mesenchymal Stem Cells (MSCs) have a high ability to renew and differentiate themselves into various lineages of conjunctive tissues. This study aimed to isolate the MSCs from murine bone marrow by using two different growth media and to characterize them with immunostaining with antivimentin antibody. We used six 2-week old BALB/c mice. Bone marrow was collected from mice's tibial and femoral channels and re-suspended in a final strength of 6x105 in Knockout-DMEM and high-glucose-DMEM media, supplemented by 10% FBS, and kept in a humidified 5% CO2 incubator at 37º C for 72 h, when non-adherent cells were removed during the change of medium. The number and density of adherent fibroblast-like colonies was greater with the Knockout-DMEM medium (within 5 days of culture) versus 10-20 days in DMEM-high glucose to get the same cellular concentration. The cells in both groups were highly positive for antivimentin antibody, characterizing them as MSCs. Obtaining MSCs as quickly as possible is essential for cell therapy field, especially when those cells are intended to be used for the repair of tissues from mesenchymal sources