Use of sisal industrial waste (Agave sisalana Perrine) in sustainable and multifunctional cosmetic products.

OBJECTIVE: Sisal is a common stiff fibre produced around the world, corresponding to approximately 70% of the commercial production of all fibres of this type. The fibres are extracted from the leaves of Agave sisalana, from which approximately 4% of their weight is obtained, with the remaining 96% considered to be residues from the process of the sisal industry. The objective of this work was to obtain a polyphenol-enriched extract from the A. sisalana residue by ultrasonically assisted extraction, characterize it chemically, evaluate in vitro antioxidant activity, and develop safe and stable photoprotective formulations for future application in cosmetic preparations. METHODS: Ultrasonic extraction of solid plant material was performed using 50% ethanol/water (v/v). The extract was chemically characterized by high-performance liquid chromatography equipment associated with classical molecular networking and evaluated for in vitro antioxidant activity by different methodologies. Ten formulations were prepared, varying the component concentrations and the shear time. The 1.0% sisal extract was incorporated into the most stable formulations, and preliminary and accelerated stability were evaluated. The emulsions were investigated for safety by assessment of primary accumulated dermal irritability and sensitization and a dermatological clinical study of phototoxicity and photosensitization. The photoprotective formulations containing or not containing the extract that were stable after 90 days had their in vivo sun protection factor (SPF), UVA protection factor, critical wavelength, and protection against visible and blue light determined. RESULTS: Ultrasound extraction using 50% ethanol/water (EH 50) as an extractor vehicle showed the best yield. The extract exhibited a concentration of phenolic compounds (77.93 mg of equivalent to the standard gallic acid/g) and showed in vitro antioxidant activity. Emulsions without and with 1.0% sisal extract remained stable and safe. The addition of the extract to the photoprotective formulation statistically increased the SPF when compared to the formulation without the extract and offered protection against UVA radiation, critical wavelengths, and absorption of visible and blue light. CONCLUSION: Based on the findings, the solid residue of A. sisalana may be indicated as a component of photoprotective and antioxidant cosmetic formulations.

OBJECTIF: Le sisal est une fibre rigide courante produite dans le monde entier, correspondant à environ 70 % de la production commerciale de toutes les fibres de ce type. Les fibres sont extraites des feuilles d'Agave sisalana dont environ 4 % du poids est obtenu, les 96 % restants étant considérés comme des résidus du procédé de l'industrie du sisal. L'objectif de ce projet était d'obtenir un extrait du résidu d'A. sisalana enrichi en polyphénols par extraction assistée par ultrasons (EAU), de le caractériser chimiquement, d'évaluer l'activité antioxydante in vitro et de développer des formulations photoprotectrices sûres et stables pour une application future dans des préparations cosmétiques. MÉTHODES: L'extraction ultrasonique de la matière végétale solide a été effectuée avec une solution à 50 % d'éthanol/eau (v/v). L'extrait a été chimiquement caractérisé avec un équipement de chromatographie en phase liquide à haute performance associé à un réseau moléculaire (RM) classique, puis évalué pour l'activité antioxydante in vitro par différentes méthodologies. Dix formulations ont été préparées en variant les concentrations des composants et le temps de cisaillement. L'extrait de sisal à 1,0 % a été incorporé dans les formulations les plus stables et la stabilité préliminaire et accélérée a été évaluée. La sécurité d'emploi des émulsions a été étudiée en évaluant l'irritabilité et la sensibilisation cutanées accumulées primaires et l'étude clinique dermatologique de la phototoxicité et de la photosensibilisation. Le facteur de protection solaire in vivo, le facteur de protection UVA, la longueur d'onde critique et la protection contre la lumière visible et bleue ont été déterminées pour les formulations photoprotectrices contenant ou non l'extrait qui étaient stables après 90 jours. RÉSULTATS: L'extraction par ultrasons utilisant une solution à 50 % d'éthanol/eau (EH 50) comme véhicule d'extraction a menée au meilleur rendement. L'extrait a présenté une concentration de composés phénoliques (77,93 mg d'EAG/g) et une activité antioxydante in vitro. Les émulsions sans et avec 1,0 % d'extrait de sisal sont restées stables et sans danger. L'ajout de l'extrait à la formulation photoprotectrice a statistiquement augmenté le SPF par rapport à la formulation sans extrait et a offert une protection contre les rayonnements UVA, la longueur d'onde critique et l'absorption de la lumière visible et bleue. CONCLUSION: D'après ces résultats, les résidus solides d'A. sisalana peuvent être indiqués comme composant des formulations cosmétiques photoprotectrices et antioxydantes.

Methyl jasmonate induces selaginellin accumulation in Selaginella convoluta.

INTRODUCTION: Selaginellins are specialized metabolites and chemotaxonomic markers for Selaginella species. Despite the growing interest in these compounds as a result of their bioactivities, they are accumulated at low levels in the plant. Hence, their isolation and chemical characterization are often difficult, time consuming, and limiting for biological tests. Elicitation with the phytohormone methyl jasmonate (MeJA) could be a strategy to increase the content of selaginellins addressing their low availability problem, that also impairs pharmacological investigations. MATHERIALS AND METHODS: In this study, we examined MeJA elicitation in Selaginella convoluta plants, a medicinal plant found in northeastern Brazil, by treating them with two different concentrations (MeJA: 50 and 100 µM), followed by chemical profiling after 12, 24 and 48 h after application. Samples were harvested and analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). RESULTS AND DISCUSSCION: MeJA treatment significantly impacted the chemical phenotype. Regarding shoots differences in the time-dependent increased accumulation of all metabolites when plants were subjected to 100 µM MeJA were observed while in roots, most metabolites had their concentrations decreased in a time-dependent fashion at the same conditions. Results support organ, MeJA concentration and time post-treatment dependence of specialized metabolite accumulation, mainly the flavonoids and selaginellins. The amount of Selaginellin G in shoots of MeJA-treated specimens increased in 5.63-fold relative to control. The molecular networking approach allowed for the putative annotation of 64 metabolites, among them, the MeJA treatment followed by targeted metabolome analysis also allowed to annotate seven unprecedented selaginellins. Additionally, the in silico bioactive potential of the annotated selaginellins highlighted targets related to neurodegenerative disorders, antiproliferative, and antiparasitic issues. Taken together, data point out MeJA exposure as a strategy to induce potentially bioactive selaginellins accumulation in S. convoluta, this approach could enable a deep investigation about the metabolic function of these metabolites in the genus as well as regarding pharmacological exploration of the undervalued potential.

Non-target molecular network and putative genes of flavonoid biosynthesis in Erythrina velutina Willd., a Brazilian semiarid native woody plant.

Erythrina velutina is a Brazilian native tree of the Caatinga (a unique semiarid biome). It is widely used in traditional medicine showing anti-inflammatory and central nervous system modulating activities. The species is a rich source of specialized metabolites, mostly alkaloids and flavonoids. To date, genomic information, biosynthesis, and regulation of flavonoids remain unknown in this woody plant. As part of a larger ongoing research goal to better understand specialized metabolism in plants inhabiting the harsh conditions of the Caatinga, the present study focused on this important class of bioactive phenolics. Leaves and seeds of plants growing in their natural habitat had their metabolic and proteomic profiles analyzed and integrated with transcriptome data. As a result, 96 metabolites (including 43 flavonoids) were annotated. Transcripts of the flavonoid pathway totaled 27, of which EvCHI, EvCHR, EvCHS, EvCYP75A and EvCYP75B1 were identified as putative main targets for modulating the accumulation of these metabolites. The highest correspondence of mRNA vs. protein was observed in the differentially expressed transcripts. In addition, 394 candidate transcripts encoding for transcription factors distributed among the bHLH, ERF, and MYB families were annotated. Based on interaction network analyses, several putative genes of the flavonoid pathway and transcription factors were related, particularly TFs of the MYB family. Expression patterns of transcripts involved in flavonoid biosynthesis and those involved in responses to biotic and abiotic stresses were discussed in detail. Overall, these findings provide a base for the understanding of molecular and metabolic responses in this medicinally important species. Moreover, the identification of key regulatory targets for future studies aiming at bioactive metabolite production will be facilitated.

Erythrina velutina Willd. alkaloids: Piecing biosynthesis together from transcriptome analysis and metabolite profiling of seeds and leaves.

Introduction: Natural products of pharmaceutical interest often do not reach the drug market due to the associated low yields and difficult extraction. Knowledge of biosynthetic pathways is a key element in the development of biotechnological strategies for plant specialized metabolite production. Erythrina species are mainly used as central nervous system depressants in folk medicine and are important sources of bioactive tetracyclic benzylisoquinoline alkaloids (BIAs), which can act on several pathology-related biological targets. Objectives: In this sense, in an unprecedented approach used with a non-model Fabaceae species grown in its unique arid natural habitat, a combined transcriptome and metabolome analyses (seeds and leaves) is presented. Methods: The Next Generation Sequencing-based transcriptome (de novo RNA sequencing) was carried out in a NextSeq 500 platform. Regarding metabolite profiling, the High-resolution Liquid Chromatography was coupled to DAD and a micrOTOF-QII mass spectrometer by using electrospray ionization (ESI) and Time of Flight (TOF) analyzer. The tandem MS/MS data were processed and analyzed through Molecular Networking approach. Results: This detailed macro and micromolecular approach applied to seeds and leaves of E. velutina revealed 42 alkaloids, several of them unique. Based on the combined evidence, 24 gene candidates were put together in a putative pathway leading to the singular alkaloid diversity of this species. Conclusion: Overall, these results could contribute by indicating potential biotechnological targets for modulation of erythrina alkaloids biosynthesis as well as improve molecular databases with omic data from a non-model medicinal plant, and reveal an interesting chemical diversity of Erythrina BIA harvested in Caatinga.

Molecular Networking Discloses the Chemical Diversity of Flavonoids and Selaginellins in Selaginella convoluta.

Selaginella convoluta is a desiccation tolerant plant native to the Brazilian semiarid region (Caatinga), endowed with an effective drought resistance mechanism. As part of our research efforts to understand the chemical diversity of S. convoluta, dehydrated (harvested in their natural habitat in the dry season) and hydrated (plant acclimated in a laboratory after rehydration) specimens were analyzed by HR-LC-ESI-MS/MS followed by a structural annotation on the Global Natural Products Social Molecular Networking Web platform. The molecular networking approach allowed for putative annotation of 39 metabolites, mainly selaginellins and flavonoids. Based on MS/MS data, three unprecedented selaginellins were annotated: 29-hydroxy selaginellin O, 29-hydroxy selaginellin A, and 4-{[2-(4-hydrophenyl)-6-[2-(4-hydroxyphenyl)ethynyl]phenyl](4-oxocyclohexa-2,5-dien-1-ylidene)methyl}benzaldehyde. Th results pointed out that valuable scientific knowledge can be obtained from studies conducted with plants in their natural habitat by allowing a more realistic profile of chemical diversity. The present study adds new information on specialized metabolites of S. convoluta, mainly flavonoids and selaginellins, and highlights the species as an untapped source of chemobiodiversity from Caatinga.

Erythroxylum pungens Tropane Alkaloids: GC-MS Analysis and the Bioactive Potential of 3-(2-methylbutyryloxy)tropan-6,7-diol in Zebrafish (Danio rerio).

Tropane alkaloids are specialized plant metabolites mostly found in the Erythroxylaceae and Solanaceae families. Although tropane alkaloids have a high degree of structural similarity because of the tropane ring, their pharmacological actions are quite distinct. Brazil is one of the main hotspots of Erythroxylum spp. diversity with 123 species (almost 66% of the species catalogued in tropical America). Erythroxylum pungens occurs in the Caatinga, a promising biome that provides bioactive compounds, including tropane alkaloids. As part of our efforts to investigate this species, 15 alkaloids in specimens harvested under different environmental conditions are presented herein. The occurrence of 3-(2-methylbutyryloxy)tropan-6,7-diol in the stem bark of plants growing in their natural habitat, greenhouse controlled conditions, and after a period of water restriction, suggests that it is a potential chemical marker for the species. This alkaloid was evaluated for several parameters in zebrafish (Danio rerio) as a model organism. Regarding toxicity, teratogenic effects were observed at 19.5 µM and the lethal dose for embryos was 18.4 µM. No mortality was observed in adults, but a behavioral screen showed psychostimulatory action at 116.7 µM. Overall, the alkaloid was able to cause zebrafish behavioral changes, prompting further investigation of its potential as a new molecule in the treatment of depression-like symptoms. In silico, targets involved in antidepressant pathways were identified by docking.

Selaginellaceae: traditional use, phytochemistry and pharmacology / Selaginellaceae: uso tradicional, fitoquímica y farmacología

Selaginella is the only genus from Selaginellaceae, and it is considered a key factor in studying evolution. The family managed to survive the many biotic and abiotic pressures during the last 400 million years. The purpose of this review is to provide an up-to-date overview of Selaginella in order to recognize their potential and evaluate future research opportunities. Carbohydrates, pigments, steroids, phenolic derivatives, mainly flavonoids, and alkaloids are the main natural products in Selaginella. A wide spectrum of in vitro and in vivo pharmacological activities, some of them pointed out by folk medicine, has been reported. Future studies should afford valuable new data on better explore the biological potential of the flavonoid amentoflavone and their derivatives as chemical bioactive entities; develop studies about toxicity and, finally, concentrate efforts on elucidate mechanisms of action for biological properties already reported.

Selaginella es el único género de Selaginellaceae, y se considera un factor clave en el estudio de la evolución. La familia logró sobrevivir a las muchas presiones bióticas y abióticas durante los últimos 400 millones de años. El propósito de esta revisión es proporcionar un resumen actualizado de Selaginella para reconocer su potencial y evaluar futuras oportunidades de investigación. Los hidratos de carbono, pigmentos, esteroides, derivados fenólicos, principalmente flavonoides, y alcaloides son los principales productos naturales en Selaginella. Se ha informado un amplio espectro de actividades farmacológicas in vitro e in vivo, algunas de ellas señaladas por la medicina popular. Los estudios futuros deberían proporcionar datos nuevos y valiosos para explorar mejor el potencial biológico de la amentoflavona flavonoide y sus derivados como entidades bioactivas químicas; desarrollar estudios sobre la toxicidad y, finalmente, concentrar los esfuerzos en dilucidar los mecanismos de acción para las propiedades biológicas ya informadas.