Multistep Optimization of β-Glucosidase Extraction from Germinated Soybeans (Glycine max L. Merril) and Recovery of Isoflavone Aglycones

Epicotyls from germinated soybeans (EGS) have great potential as sources of endogenous β-glucosidase. Furthermore, this enzyme may improve the conversion of isoflavones into their corresponding aglycones. β-Glucosidase may also increase the release of aglycones from the cell wall of the plant materials. Therefore, the aim of this work was to optimize both the extraction of β-glucosidase from EGS and to further examine its application in defatted soybean cotyledon to improve the recovery of aglycones, which were evaluated by ultra-high performance liquid chromatography (UHPLC). A multistep optimization was carried out and the effects of temperature and pH were investigated by applying a central composite design. The linear effect of pH and the quadratic effect of pH and temperature were significant for the extraction of β-glucosidase and recovery aglycones, respectively. Optimum extraction of β-glucosidase from EGS occurred at 30 °C and pH 5.0. Furthermore, the maximum recovery of aglycones (98.7%), which occurred at 35 °C and pH 7.0–7.6 during 144 h of germination, increased 8.5 times with respect to the lowest concentration. The higher bioaccessibility of aglycones when compared with their conjugated counterparts is well substantiated. Therefore, the data provided in this contribution may be useful for enhancing the benefits of soybean, their products, and/or their processing by-products

DESENVOLVIMENTO E VALIDAÇÃO DE MÉTODO ESPECTROFOTOMÉTRICO PARA DETERMINAÇÃO QUANTITATIVA DO ANTIRRETROVIRAL TENOFOVIR

O fumarato de tenofovir desoproxila (TDF), pró-fármaco oral, pertencente à classe dos inibidores da transcriptase reversa análogos de nucleosídeos (ITRNs). Os ITRNs destacam-se como terapêutica de primeira linha contra a infecção pelo HIV-1, sendo o TDF um dentre os produzidos no Brasil. Visando uma produção de medicamentos que assegure o cumprimento de padrões de qualidade, garantindo eficácia e segurança, o controle de qualidade dos produtos farmacêuticos necessita de métodos analíticos validados. Com esse objetivo, um método espectrofotométrico para a determinação quantitativa do FTD na matéria-prima e comprimidos utilizando HCl 0,10 mol L-1 como solvente a 258 nm foi desenvolvido e validado. Não houve interferência dos excipientes e revestimento testados. A curva de calibração, obtida a partir da relação absorvância versus concentração, mostrou-se linear na faixa de 4,00 a 38,00 mg mL-1 apresentando Equação da reta (y= 0.0213x + 0.0271) com coeficiente de determinação de 0,995. A exatidão média do método para três níveis de concentração foi de 99,0% (DPR=0,66 %) com precisão repetitividade e intermediária adequadas (DPR<5%) e limites de detecção de 2,52 mg mL-1 e de quantificação de  8,70 mg mL-1. Assim, este método constitui uma ferramenta analítica importante, por ser simples, sendo útil para o controle de qualidade do antirretroviral tenofovir, já que esse é um dos medicamentos mais caro do esquema terapêutico, distribuído gratuitamente no Brasil para portadores do vírus da AIDS e da Hepatite B

Kaurenoic Acid Possesses Leishmanicidal Activity by Triggering a NLRP12/IL-1β/cNOS/NO Pathway

Leishmania amazonensis (L. amazonensis) infection can cause severe local and diffuse injuries in humans, a condition clinically known as American cutaneous leishmaniasis (ACL). Currently, the therapeutic approach for ACL is based on Glucantime, which shows high toxicity and poor effectiveness. Therefore, ACL remains a neglected disease with limited options for treatment. Herein, the in vitro antiprotozoal effect and mechanisms of the diterpene kaurenoic acid [ent-kaur-16-en-19-oic acid] (KA) against L. amazonensis were investigated. KA exhibited a direct antileishmanial effect on L. amazonensis promastigotes. Importantly, KA also reduced the intracellular number of amastigote forms and percentage of infected peritoneal macrophages of BALB/c mice. Mechanistically, KA treatment reestablished the production of nitric oxide (NO) in a constitutive NO synthase- (cNOS-) dependent manner, subverting the NO-depleting escape mechanism of L. amazonensis. Furthermore, KA induced increased production of IL-1β and expression of the inflammasome-activating component NLRP12. These findings demonstrate the leishmanicidal capability of KA against L. amazonensis in macrophage culture by triggering a NLRP12/IL-1β/cNOS/NO mechanism

Kaurenoic Acid Possesses Leishmanicidal Activity by Triggering a NLRP12/IL-1 /cNOS/NO Pathway

Leishmania amazonensis (L. amazonensis) infection can cause severe local and diffuse injuries in humans, a condition clinically known as American cutaneous leishmaniasis (ACL). Currently, the therapeutic approach for ACL is based on Glucantime, which shows high toxicity and poor effectiveness. Therefore, ACL remains a neglected disease with limited options for treatment. Herein, the in vitro antiprotozoal effect and mechanisms of the diterpene kaurenoic acid [ent-kaur-16-en-19-oic acid] (KA) against L. amazonensis were investigated. KA exhibited a direct antileishmanial effect on L. amazonensis promastigotes. Importantly, KA also reduced the intracellular number of amastigote forms and percentage of infected peritoneal macrophages of BALB/c mice. Mechanistically, KA treatment reestablished the production of nitric oxide (NO) in a constitutive NO synthase-(cNOS-) dependent manner, subverting the NO-depleting escape mechanism of L. amazonensis. Furthermore, KA induced increased production of IL-1 and expression of the inflammasome-activating component NLRP12. These findings demonstrate the leishmanicidal capability of KA against L. amazonensis in macrophage culture by triggering a NLRP12/IL-1 /cNOS/NO mechanism