Antifungal activity of eukaryotic microalgae extracts in dermatophytes

Introduction: Microalgae are considered a promising source of biologically active secondary metabolites, with several biotechnological properties already recorded, including antifungal activity. Objective: Thus, the properties of ethanol extracts of microalgae Ankistrodesmus falcatus, Chaetoceros neogracilis, Desmodesmus brasiliensis, Dunaliella tertiolecta, Kirchneriella lunaris and Tetraselmis gracilis were investigated in vitro for antifungal activity against dermatophytes Nannizzia gypsea, Nannizzia nana, Trichophyton mentagrophytes and Trichophyton tonsurans. Methods: The extracts were evaluated using broth microdilution methodology, with a test interval of 0.0115 to 6 mg mL-1. The antifungal itraconazole was tested at concentrations 0.0313 to 16 μg mL-1. Results: All microalgae extracts showed antifungal activity, especially extracts from C. neogracilis, D. brasiliensis and K. lunaris that totally inhibited growth of all species of dermatophytes evaluated. The lowest MIC values recorded were for the extracts of D. brasiliensis and K. lunaris (MIC 0.188 mg mL-1), against T. tonsurans e N. nana. Conclusions: This was a pioneering work about the antifungal activity of these microalgae against dermatophytes. It is expected that, from this study, further research will be carried out in order to identify and isolate the active biomolecules responsible for the antifungal activity of these microalgae species.Introdução: as microalgas são consideradas uma fonte promissora de metabólitos secundários biologicamente ativos, com diversas propriedades biotecnológicas já registradas, incluindo atividade antifúngica. Objetivo: assim, as propriedades dos extratos etanólicos das microalgas Ankistrodesmus falcatus, Chaetoceros neogracilis, Desmodesmus brasiliensis, Dunaliella tertiolecta, Kirchneriella lunaris e Tetraselmis gracilis foram investigadas in vitro quanto à atividade antifúngica frente aos dermatófitos Nannizzia gypsea, Nannizzia nana, Trichophyton mentagrophytes e Trichophyton tonsurans. Metodologia: os extratos foram avaliados através da metodologia de microdiluição em caldo, com intervalo teste de 11.5 a 6.000 μg mL-1. O antifúngico itraconazol foi testado nas concentrações 0.0313 a 16 μg mL-1. Resultados: todos os extratos microalgais apresentaram atividade antifúngica, com destaque para os extratos das microalgasC. neogracilis, D. brasiliensis e K. lunaris, que inibiram totalmente o crescimento de todas as espécies de dermatófitos avaliadas. Os menores valores de CIM registrados foram para os extratos de D. brasiliensis e K. lunaris (CIM 188 μg mL-1), contra T. tonsurans e N. nana. Conclusão: este foi um trabalho pioneiro acerca da atividade antifúngica destas microalgas frente à dermatófitos. Espera-se que, a partir deste estudo, novas pesquisas sejam realizadas a fim de identificar e isolar as biomoléculas ativas responsáveis pela atividade antifúngica destas espécies de microalgas

Caracterização da enzima lisina cetoglutarato redutase (LKR)/ sacaropina desidrogenase (SDH) estudada em Phaseolus vulgaris

Orientadores: Luiz Gonzaga Santoro, Paulo ArrudaTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A via de degradação da lisina em plantas é catalizada pela enzima bifuncional lisina cetoglutarato redutase (LKR) / sacaropina desidrogenase (SOH). A LKR condensa lisina e a-cetoglutarato em sacaropina usando NAOPH como cofator e a SOH converte a sacaropina em a-aminoadipato-õ-semialdeido e ácido glutâmico, usando NAO+ como cofator. No presente trabalho foi demonstrado, em Phaseo/us vu/garis, a participação desta enzima no catabolismo de lisina. A atividade foi medida em diferentes tecidos desta espécie. O hipocótilo apresentou maior atividade específica em plantas estioladas, seguidas pela folha, vagem e raiz, com valores muito inferiores. Sementes e cotilédones não demonstraram presença mensurável da enzima. A LKRlSOH também foi encontrada em plântulas de feijão carioca 80 crescidas in vitro à partir de sementes cujo cotilédone foi retirado previamente. Usando o sistema in vitro, a resposta enzimática à adição de lisina no meio de crescimento foi testada, demonstrando que o substrato não causa alterações na atividade. A precipitação de proteínas do tecido de hipocótilo em concentrações crescentes de PEG 8000 revelou dois platõs de atividade, levantando a hipótese de haver duas isoformas da enzima. Essa possibilidade foi reforçada posteriormente devido à respostas diferentes à inibidores de fosfatase, os quais revelaram que a LKRlSOH em feijão é uma fosfoproteína, assim como já foi observado em outras plantas. A enzima do hipocótilo foi isolada por métodos cromatográficos, indicando ser um polipeptídeo bifuncional. No entanto, dependendo do procedimento de purificação, a enzima pode eluir como um monõmero de 94kOa, contendo apenas a atividade da SOH, ou como um dímero de 190kOa, com ambas atividades eluindo conjuntamente. As condições de iluminação durante o crescimento da planta revelaram exercer influência na atividade da LKRlSOHAbstract: The pathway of Iysine catabolism in plants is catalyzed by the bifunctional enzyme lysine ketogltutarate reductase (LKR) /saccharopine dehydrogenase (SOH). LKR condenses lysine and "alfa" ketoglutarate into saccharopine, using NAOPH as a cofactor and SOH converts saccharopine into a-aminoadipate õ-semialdehyde and glutamic acid, using NAO+ as a cofactor. In the present work it was demonstrated, in Phaseolus vulgaris , the participation of this enzyme in lysine catabolism. The activity was measured in different tissues from this species. The hipocotyl presented the highest specific activity in etiolated plants, followed by the leaf, pod and root, with much lower values. Seeds and cotyledons did not present any measurable activity. LKRlSOH was also found in carioca 80 seedlings grown, in vitro, from seeds whose cotyledon was previously removed. Using the in vitro system, the response to Iysine in the growing medium was tested revealing that the substrate didn't result in any change in enzyme activity. The precipitation of proteins from hipocotyl tissue with increasing concentrations of PEG 8000 revealed two p/atos of enzyme activity, raising the hypothesis that two isoforms of this enzyme are present. This possibility was subsequently reinforced due to different responses to phosphatase inhibitors, which revealed that LKRlSOH in P. vulgaris is a phosphoprotein, as observed with other plants. The hipocotyl enzyme was isolated by chromatographic methods, showing it to be a bifunctional polypeptide. However, depending on the purification procedure, it may elute as a monomer of 94 kOa containing only SOH activity, or as a dimer of 190 kOa where both activities elute together. Light conditions during plant growth were shown to influence the activity of LKRlSOHDoutoradoBiologia VegetalDoutor em Ciência

Ultraestrutura celular e expressão de proteínas de leveduras hanseniaspora sob efeito do estresse etanólico

Resumo O objetivo deste estudo foi avaliar a resposta de Hanseniaspora opuntiae (Ho41) e H. guilliermondii (Hg43) ao estresse etanólico, observando a ultraestrutura e o perfil de expressão proteica em concentrações crescentes de etanol. A ultraestrutura foi analisada por microscopia eletrônica de varredura (MEV) e a expressão proteica, pelo perfil eletroforético (SDS-PAGE). Na análise microscópica, as cepas em meio Yeast Malt Agar sem etanol mostraram células jovens com morfologia apiculada, brotamento bilateral e polos distais côncavos. Com o início do estresse, a 3% de etanol, as células apresentaram múltiplas cicatrizes em forma de anéis e, com 6%, alterações na integridade da parede celular, plasmólise e ativação da autólise. Na análise eletroforética, observou-se, tanto para Ho41 quanto para Hg43, aumento na expressão de um peptídeo de 100 kDa, com aumento do etanol no meio, indicando ser uma proteína de choque térmico (HSP). As HSPs vêm sendo patenteadas como marcadores de organismos de interesse biotecnológico, já que as condições necessárias para obtenção de bioprodutos muitas vezes requerem cultivo sob estresse. Neste contexto, esta proteína pode ser indicada como marcador molecular para bioprospecção ou melhoramento genético de cepas não-saccharomyces mais resistentes aos processos de fermentação, na fabricação de vinhos

Estrogen and Thyroid Hormone Receptor Activation by Medicinal Plants from Bahia, Brazil

Background: A number of medicinal plants are traditionally used for metabolic disorders in Bahia state, Brazil. The aim of this study was to evaluate the estrogen receptor (ER) and thyroid receptor (TR) activation of crude extracts prepared from 20 plants. Methods: Species were extracted and assayed for receptor activation through both ER and TR gene-reporter assays, using 17β-estradiol and triiodothyronine (T3), respectively, as the positive controls. Results: Cajanus cajan (Fabaceae), Abarema cochliacarpus (Fabaceae), and Borreria verticillata (Rubiaceae) were able to activate ER as much as the positive control (17β-estradiol). These three plant species were also assayed for TR activation. At the concentration of 50 µg/mL, C. cajans exerted the highest positive modulation on TR, causing an activation of 59.9%, while B. verticillata and A. cochliacarpus caused 30.8% and 23.3%, respectively. Conclusions: Our results contribute towards the validation of the traditional use of C. cajans, B. verticillata, and A. cochliacarpus in the treatment of metabolic disorders related to ER and TR functions. The gene-reporter assay was proven effective in screening crude plant extracts for ER/TR activation, endorsing this methodology as an important tool for future bioprospection studies focused on identifying novel starting molecules for the development of estrogen and thyroid agonists

Implementation of a Brazilian Cardioprotective Nutritional (BALANCE) Program for improvement on quality of diet and secondary prevention of cardiovascular events: A randomized, multicenter trial

Background: Appropriate dietary recommendations represent a key part of secondary prevention in cardiovascular disease (CVD). We evaluated the effectiveness of the implementation of a nutritional program on quality of diet, cardiovascular events, and death in patients with established CVD. Methods: In this open-label, multicenter trial conducted in 35 sites in Brazil, we randomly assigned (1:1) patients aged 45 years or older to receive either the BALANCE Program (experimental group) or conventional nutrition advice (control group). The BALANCE Program included a unique nutritional education strategy to implement recommendations from guidelines, adapted to the use of affordable and regional foods. Adherence to diet was evaluated by the modified Alternative Healthy Eating Index. The primary end point was a composite of all-cause mortality, cardiovascular death, cardiac arrest, myocardial infarction, stroke, myocardial revascularization, amputation, or hospitalization for unstable angina. Secondary end points included biochemical and anthropometric data, and blood pressure levels. Results: From March 5, 2013, to Abril 7, 2015, a total of 2534 eligible patients were randomly assigned to either the BALANCE Program group (n = 1,266) or the control group (n = 1,268) and were followed up for a median of 3.5 years. In total, 235 (9.3%) participants had been lost to follow-up. After 3 years of follow-up, mean modified Alternative Healthy Eating Index (scale 0-70) was only slightly higher in the BALANCE group versus the control group (26.2 ± 8.4 vs 24.7 ± 8.6, P <.01), mainly due to a 0.5-serving/d greater intake of fruits and of vegetables in the BALANCE group. Primary end point events occurred in 236 participants (18.8%) in the BALANCE group and in 207 participants (16.4%) in the control group (hazard ratio, 1.15; 95% CI 0.95-1.38; P =.15). Secondary end points did not differ between groups after follow-up. Conclusions: The BALANCE Program only slightly improved adherence to a healthy diet in patients with established CVD and had no significant effect on the incidence of cardiovascular events or death. © 2019 The Author