Secure and Sustainable Sourcing of Plant Tissues for the Exhaustive Exploration of Their Chemodiversity.

The main challenge of plant chemical diversity exploration is how to develop tools to study exhaustively plant tissues. Their sustainable sourcing is a limitation as bioguided strategies and dereplication need quite large amounts of plant material. We examine if alternative solutions could overcome these difficulties by obtaining a secure, sustainable, and scalable source of tissues able to biosynthesize an array of metabolites. As this approach would be as independent of the botanical origin as possible, we chose eight plant species from different families. We applied a four steps culture establishment procedure, monitoring targeted compounds through mass spectrometry-based analytical methods. We also characterized the capacities of leaf explants in culture to produce diverse secondary metabolites. In vitro cultures were successfully established for six species with leaf explants still producing a diversity of compounds after the culture establishment procedure. Furthermore, explants from leaves of axenic plantlets were also analyzed. The detection of marker compounds was confirmed after six days in culture for all tested species. Our results show that the first stage of this approach aiming at easing exploration of plant chemodiversity was completed, and leaf tissues could offer an interesting alternative providing a constant source of natural compounds.

Allantoin reduces cell death induced by cisplatin: possible implications for tumor lysis syndrome management.

Massive lysis of tumor mass in cancer patients under chemotherapy regimens generates high levels of uric acid, leading to what is known as tumor lysis syndrome (TLS). Rasburicase, a recombinant urate oxidase, converts urate to allantoin, which is readily excreted by the kidneys. Even though there is a high production of allantoin from urate in cancer patients following rasburicase treatment, there are no studies on how allantoin excess could interfere with chemotherapy. We have evaluated allantoin interference with cisplatin efficiency on the lung cancer cell line H460 in vitro. The cells were treated with cisplatin (33 µM), with or without allantoin, for 48 h, in the presence or absence of UV light, and N-acetyl-L-cysteine (NAC) for 24 h. Cell viability, cell cycle, ROS production, apoptosis and immunoblot assays were performed. We showed that allantoin reduced the apoptosis induced by cisplatin in the H460 cell line. However, the activity of carboplatin and oxaliplatin, betulinic acid, TIBA, UV and H2O2 was not affected by allantoin. NMR spectroscopy showed that allantoin reduces cisplatin activity through direct interaction with cisplatin.

Using LC and Hierarchical Cluster Analysis as Tools to Distinguish Timbó Collections into Two Deguelia Species: A Contribution to Chemotaxonomy.

The species Deguelia utilis and Deguelia rufescens var. urucu, popularly known as "timbó," have been used for many years as rotenone sources in insecticide formulations. In this work, a method was developed and validated using a high-performance liquid chromatography-photodiode array (HPLC-PDA) system, and results were analyzed using hierarchical cluster analysis (HCA). By quantifying the major rotenoids of these species, it was possible to establish a linear relation between them. The ratio between the concentrations of rotenone and deguelin for D. utilis is approximately 1:0.8, respectively, while for D. rufescens var. urucu it is 2:1. These results may help to distinguish these species contributing to their taxonomic identification.

Flavonoids from leaves of Derris urucu: assessment of potential effects on seed germination and development of weeds.

In some previous studies, we described the isolation of nine compounds from leaves of Derris urucu, a species found widely in the Amazon rainforest, identified as five stilbenes and four dihydroflavonols. In this work, three of these dihydroflavonols [urucuol A (1), urucuol B (2) and isotirumalin (3)] were evaluated to identify their potential as allelochemicals, and we are also reporting the isolation and structural determination of a new flavonoid [5,3'-dihydroxy-4'-methoxy-(7,6:5″,6″)-2″,2″-dimethylpyranoflavanone (4)]. We investigated the effects of the dihydroflavonols 1-3 on seed germination and radicle and hypocotyl growth of the weed Mimosa pudica, using solutions at 150 mg.L-1. Urucuol B, alone, was the substance with the greatest potential to inhibit seed germination (26%), while isotirumalin showed greater ability to reduce the development of the hypocotyl (25%), but none of the three substances showed the potential to inhibit radicle. When combined in pairs, the substances showed synergism for the development of root and hypocotyl and effects on seed germination that could be attributed to antagonism. When tested separately, the trend has become more intense effects on seed germination, while for the substances tested in pairs, the intensity of the effect was greater on development of weed.

Pellucidin A promotes antinociceptive activity by peripheral mechanisms inhibiting COX-2 and NOS: In vivo and in silico study.

Peperomia pellucida (PP) belongs to the Peperomia genus, which has a pantropic distribution. PP is used to treat a wide range of symptoms and diseases, such as pain, inflammation, and hypertension. Intriguingly, PP extract is used by different tropical countries for its anti-inflammatory and antinociceptive effects. In fact, these outcomes have been shown in animal models, though the exact bioactive products of PP that exert such results are yet to be discovered. To determine and elucidate the mechanism of action of one of these compounds, we evaluated the antinociceptive effect of the novel dimeric ArC2 compound, Pellucidin A by using in vivo and in silico models. Animals were then subjected to chemical, biphasic and thermal models of pain. Pellucidin A induced an antinociceptive effect against chemical-induced pain in mice, demonstrated by the decrease of the number of writhes, reaching a reduction of 43% and 65% in animals treated with 1 and 5 mg/kg of Pellucidin A, respectively. In the biphasic response (central and peripheral), animals treated with Pellucidin A showed a significant reduction of the licking time exclusively during the second phase (inflammatory phase). In the hot-plate test, Pellucidin A did not have any impact on the latency time of the treated animals. Moreover, in vivo and in silico results show that Pellucidin A's mechanism of action in the inflammatory pain occurs most likely through interaction with the nitric oxide (NO) pathway. Our results demonstrate that the antinociceptive activities of Pellucidin A operate under mechanism(s) of peripheral action, involving inflammatory mediators. This work provides insightful novel evidence of the biological properties of Pellucidin A, and leads to a better understanding of its mechanism of action, pointing to potential pharmacological use.

Effects of Rosmarinus officinalis essential oil on germ tube formation by Candida albicans isolated from denture wearers.

INTRODUCTION: The aim of this study was to investigate the effects of Rosmarinus officinalis essential oil on germ tube formation by Candida albicans isolated from denture wearers. METHODS: Ten C. albicans isolates recovered from denture wearers were tested using 10% fetal bovine serum with or without 4% R. officinalis essential oil. RESULTS: The essential oil from R. officinalis completely inhibited germ tube formation in the investigated C. albicans isolates. CONCLUSIONS: The results demonstrate that the essential oil of R. officinalis modulates C. albicans pathogenicity through its primary virulence factor (i.e., germ tube formation was suppressed).

Chemical constituents and allelopathic and antioxidant activities of Alchorneopsis floribunda Müll. Arg. (Euphorbiaceae).

Chemical investigation of extracts from the stems and leaves of Alchorneopsis floribunda Müll. Arg., collected in the Amazon region, was performed. The main isolated compounds were triterpenes (α-amyrin, ß-amyrin, lupeol, betulin, betulinic acid, uvaol, erythrodiol and oleanolic acid) and phenolic acid derivatives from 4-hydroxybenzoic acid (gallic and protocatechuic acids and isocorilagin). In the germination assays, high inhibitory allelopathic effects of the extracts and isocorilagin were observed and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of isocorilagin was higher than those of the standards used (Trolox and butylated hydroxyanisole). This is the first chemical study of the genus Alchorneopsis (Euphorbiaceae).

Reflexões Teórico-Metodológicas acerca da Pesquisa em Compreensão de Textos com Crianças / Theoretical-Methodological Discussion on Research in Reading Comprehension with Children

RESUMO Dada sua natureza multifacetada, a compreensão de textos tem sido investigada a partir de uma grande variedade de recursos metodológicos. O presente artigo tece uma reflexão de natureza teórico-metodológica acerca dos principais métodos de investigação adotados nas pesquisas realizadas com crianças sobre esse tema. A análise recai sobre as principais características desses métodos, seus limites e possibilidades que apresentam ao examinar diferentes aspectos envolvidos na compreensão de textos. Dois parâmetros foram considerados: as unidades linguísticas investigadas e o momento em que a compreensão é avaliada. As análises e discussões conduzidas ilustram a estreita relação entre método e dado, contribuindo para que o pesquisador tenha uma visão mais clara daquilo que os métodos efetivamente permitem examinar sobre a compreensão de textos.

ABSTRACT Due to its multifaceted nature, reading comprehension has been investigated from a variety of methods. This paper focuses on a theoretical-methodological discussion about the methods usually adopted in studies with children. The analysis focused on the main characteristics of these methods, their limits, and possibilities they present in examining different aspects involved in reading comprehension. Two parameters were considered: the linguistic units investigated, and the moment at which comprehension is evaluated. The analyses and discussions conducted illustrate the close relationship between method and data, helping the researcher to have a clearer view of what methods effectively allow the examination about text comprehension.

Biflavones and triterpenoids isolated from Ouratea castaneifolia (DC.) Engl., Ochnaceae / Biflavonas e triterpenóides isolados de Ouratea castaneifolia (DC.) Engl., Ochnaceae

This paper presents the chemical investigation of the leaves and stems of Ouratea castaneifolia (DC.) Engl.. There are no chemical or pharmacological studies with this species. Classic phytochemical investigation of the organic extracts together with high pressure liquid chromatography (HPLC) procedures lead to the identification of seventeen metabolites: seven triterpenes (friedelin, 3β-friedelinol, α-amyrin, β-amyrin, lupeol, germanicol and taraxerol), four steroids (sitosterol, stigmasterol and the glycosides sitosteryl 3-O-β-D-glucopyranoside and stigmasteryl 3-O-β-D-glucopyranoside), one isoflavone (5,7,4'-trimethoxyisoflavone), one flavone (5,4'-dihydroxy-7,3',5'-trimethoxyflavone) and four biflavones (amenthoflavone, 7,7"-O-dimethylamenthoflavone, heveaflavone and tetramethylamenthoflavone). The structures of the compounds were established by the analysis of ¹H, 13C NMR spectra including bidimensional techniques. The classes of the identified metabolites are in agreement with previous studies of the Ouratea genus.

O presente trabalho trata da investigação química das folhas e caule da espécie Ouratea castaneifolia (DC.) Engl., sobre a qual não há registros de estudos químicos ou farmacológicos anteriores. O estudo fitoquímico clássico dos extratos orgânicos do caule e das folhas de O. castaneifolia foi aliado à técnica da cromatografia líquida de alta eficiência (CLAE) e resultou na identificação de dezessete metabólitos: sete triperpenos (friedelina, 3β-friedelinol, α-amirina, β-amirina, lupeol, taraxerol e germanicol), quatro esteróides (sitosterol, estigmasterol e os glucosídeos sitosteril 3-O-β-D-glicopiranosídeo e estigmasteril 3-O-β-D-glicopiranosídeo), uma isoflavona (5,7,4´-trimetoxiisoflavona), uma flavona (5,4´-diidroxi-7,3´,5´-trimetoxiflavona), quatro biflavonas (amentoflavona, 7,7"-O-dimetil-amentoflavona, heveaflavona e tetrametilamentoflavona). A identificação das substâncias foi feita com base na análise de espectros de RMN de ¹H, 13C e técnicas bidimensionais. As classes dos metabólitos identificados estão de acordo com aquelas citadas em estudos químicos do gênero Ouratea.

Antraquinonas e naftoquinonas do caule de um espécime de reflorestamento de Tectona grandi (Verbenaceae) / Anthraquinones and naphtoquinones from the bark of Tectona grandis (Verbenaceae) reforestation specimen

O fracionamento do extrato hexânico do caule de um espécime de reflorestamento de Tectona grandis (Verbenaceae), através de procedimentos fitoquímicos clássicos, levou ao isolamento das naftoquinonas lapachol e desidro-a-lapachona e das antraquinonas tectoquinona e obtusifolina. As estruturas das substâncias foram caracterizadas através da análise de métodos espectrométricos de RMN. Este é o primeiro estudo fitoquímico de um espécime de reflorestamento de Tectona grandis, no Brasil, sendo o objetivo principal deste trabalho a comprovação da presença de tectoquinona em espécimes cultivados.

The hexane extract of the bark of Tectona grandis (Verbenaceae) afforded two anthraquinones and two naphtoquinones. Their caracterizations were obtained through NMR spectroscopic techniques. This is the first phytochemical study of the bark of Tectona grandis reforestation specimen in Brazil. The main interest in this work is proving the presence of tectoquinone in reforestation specimen.