Detection of isoflavones and phytoestrogen-rich plant extracts binding to estrogen receptor ß using a yeast-based fluorescent assay.

Alchemilla vulgaris L., Trifolium pratense L. and Glycyrrhiza glabra L. are important remedies in traditional medicine, known for many usages, including treating gynecological diseases. Despite folkloric use of the plant materials, there is a lack of scientific data to support their therapeutic application. The aims of the present study were to evaluate the relative binding affinities (RBAs) of plant-derived phytoestrogens for estrogen receptor ß (ERß) using fluorescent biosensor in yeast and to apply this assay for the assessment of the potential of plant materials towards ERs and treatment of estrogen-related disorders. Ligand-binding domain of ERß fused with yellow fluorescent protein (ERß LBD-YFP) was expressed in S. cerevisiae and fluorescence was detected by fluorimetry and fluorescence microscopy. Structural basis for experimental results was explored by molecular docking. Yeast-based fluorescent assay was successfully optimized and applied for identification of natural phenolic compounds and phytoestrogen-rich plant extracts that interact with ERß-LBD, making this biosensor a valuable tool for screening estrogenic potential of a variety of plant extracts. This assay can be used for preliminary testing of plant-derived or fungal extracts, but also other sources of environmental substances with ER-modulating activity in order to assess their possible effects on the female reproductive system.

Anti-Inflammatory, Antioxidant and Enzyme Inhibition Activities in Correlation with Mycochemical Profile of Selected Indigenous Ganoderma spp. from Balkan Region (Serbia).

The aim of this research work was to study the bioactivity potentials (antioxidant, anti-inflammatory, and enzyme inhibitory) of ethanol (EtOH), water (H2 O) and chloroform (CHCl3 ) extracts of G. applanatum, G. lucidum, G. pfeifferi and G. resinaceum as well as their mycochemical profile: the total content of phenolics (TP) and sugars (TS) and LC/MS/MS detection of phenolics. LC/MS/MS profile showed that p-hydroxybenzoic and protocatechuic acids were mostly found. The highest ABTS and DPPH activities were detected in polar G. applanatum extracts (159.84±0.59 mg TE/g d.w., IC50 =0.85±0.30 µg/mL, respectively), while G. resinaceum CHCl3 extract was the most potent in NO assay (IC50 =41.21±0.18-81.89±0.81 µg/mL). The highest TP and TS were generally determined in G. applanatum EtOH extracts. Enzyme inhibitory effects were determined in H2 O extracts. Generally, CHCl3 extracts showed the most powerful anti-inflammatory potential. These results suggest that analyzed species are a promising source of bioactive compounds and may be considered as candidates for new food supplements or drug formulations.

Comparative In Vitro Antioxidant Capacity and Terpenoid Profiling of Pumpkin Fruit Pulps from a Serbian Cucurbita maxima and Cucurbita moschata Breeding Collection.

Pumpkin is considered a healthy and functional food. The consumption of pumpkins and pumpkin-based foods has been shown to confer several beneficial effects on human health due to their antioxidant capacity and terpenoid content. Consequently, this study aimed to characterize the in vitro antioxidant capacity (using FRAP and ABTS assays), terpenoid profile (using an untargeted lipidomics approach via high-resolution UHPLC-Orbitrap mass spectrometry), and carotenoid content (by HPLC-DAD) in pumpkin fruit pulp from accessions differing for species (11 Cucurbita maxima and 9 Cucurbita moschata), cultivar, and origin, belonging to a Serbian breeding collection. These accessions are candidates for inclusion within programs intended to improve pumpkin fruit quality. The results obtained in this work allowed us to highlight the best marker compounds, discriminating both the region of accession collection or breeding ("origin") and the plant species. Furthermore, our findings have helped to identify the most suitable antioxidant-rich varieties to select for national breeding programs for improving human health. These findings provide valuable information to the overall current understanding of the potential health benefits of pumpkins and the discriminant triterpenoids underlying the C. maxima and C. moschata accessions investigated here, which include those of Serbian and non-Serbian origin.