Synthesis of Thiazolidin-4-Ones Derivatives, Evaluation of Conformation in Solution, Theoretical Isomerization Reaction Paths and Discovery of Potential Biological Targets.

Thiazolin-4-ones and their derivatives represent important heterocyclic scaffolds with various applications in medicinal chemistry. For that reason, the synthesis of two 5-substituted thiazolidin-4-one derivatives was performed. Their structure assignment was conducted by NMR experiments (2D-COSY, 2D-NOESY, 2D-HSQC and 2D-HMBC) and conformational analysis was conducted through Density Functional Theory calculations and 2D-NOESY. Conformational analysis showed that these two molecules adopt exo conformation. Their global minimum structures have two double bonds (C=N, C=C) in Z conformation and the third double (C=N) in E. Our DFT results are in agreement with the 2D-NMR measurements. Furthermore, the reaction isomerization paths were studied via DFT to check the stability of the conformers. Finally, some potential targets were found through the SwissADME platform and docking experiments were performed. Both compounds bind strongly to five macromolecules (triazoloquinazolines, mglur3, Jak3, Danio rerio HDAC6 CD2, acetylcholinesterase) and via SwissADME it was found that these two molecules obey Lipinski's Rule of Five.

Phytochemical Study on Seeds of Paeonia clusii subsp. rhodia-Antioxidant and Anti-Tyrosinase Properties.

In this study, the black fertile (BSs) and the red unfertile seeds (RSs) of the Greek endemic Paeonia clusii subsp. rhodia (Stearn) Tzanoud were studied for the first time. Nine phenolic derivatives, trans-resveratol, trans-resveratrol-4'-O-ß-d-glucopyranoside, trans-ε-viniferin, trans-gnetin H, luteolin, luteolin 3'-O-ß-d-glucoside, luteolin 3',4'-di-O-ß-d-glucopyranoside, and benzoic acid, along with the monoterpene glycoside paeoniflorin, have been isolated and structurally elucidated. Furthermore, 33 metabolites have been identified from BSs through UHPLC-HRMS, including 6 monoterpene glycosides of the paeoniflorin type with the characteristic cage-like terpenic skeleton found only in plants of the genus Paeonia, 6 gallic acid derivatives, 10 oligostilbene compounds, and 11 flavonoid derivatives. From the RSs, through HS-SPME and GC-MS, 19 metabolites were identified, among which nopinone, myrtanal, and cis-myrtanol have been reported only in peonies' roots and flowers to date. The total phenolic content of both seed extracts (BS and RS) was extremely high (up to 289.97 mg GAE/g) and, moreover, they showed interesting antioxidative activity and anti-tyrosinase properties. The isolated compounds were also biologically evaluated. Especially in the case of trans-gnetin H, the expressed anti-tyrosinase activity was higher than that of kojic acid, which is a well-known whitening agent standard.

Heliotropium procubens Mill: Taxonomic Significance and Characterization of Phenolic Compounds via UHPLC-HRMS- In Vitro Antioxidant and Enzyme Inhibitory Activities.

The aim of the present study was the phytochemical analysis of the aerial parts of Heliotropium procumbens Mill., a herb from Boraginaceae plant family not previously studied. The methanol (ME) and aqueous extracts (WE) of the aerial parts were assayed for their total phenolic and flavonoid content and antioxidant properties, using free radical scavenging (DPPH, ABTS), reducing power (FRAP, CUPRAC), phosphomolybdenum and metal chelating assays. The extracts displayed considerable free radical scavenging activity against DPPH and ABTS radicals, with potential values of 46.88 and 68.31 mg TE/g extract for ME, and 93.43 and 131.48 mg TE/g extract for WE, respectively. Key clinical enzymes involved in neurodegenerative diseases AChE and BChE, diabetes (α-amylase and α-glucosidase) and skin whitening (tyrosinase) were also assayed. The phytochemical profile of the studied species was determined through UHPLC-HRMS, whereby 26 secondary metabolites were identified, three of which (luteolin-7-glucoside, lithospermic and rosmarinic acids) were isolated and structurally determined by NMR spectral means. H. procubens was found to harbor bioactive metabolites and could, hence, serve as a source of biological activities which could be further explored and exploited for potential applications.

Assessment of Antioxidant and Antimicrobial Properties of Selected Greek Propolis Samples (North East Aegean Region Islands).

Propolis is a bee-produced substance rich in bioactive compounds, which has been utilized widely in folk medicine, in food supplement and cosmetology areas because of its biological properties, (antibacterial, antiviral, antioxidant, anti-inflammatory, etc.). The subject of this study is associated with the chemical analysis and the biological evaluation of 16 propolis samples from the northeast Aegean region Greek islands, a well-recognized geographic area and the homeland of rich flora as a crossroads between Europe and Asia. Our study resulted in the detection of a significant percentage of diterpenes by gas chromatography-mass spectrometry (GC-MS), while flavonoids were identified in low percentages among studied samples. Furthermore, the DPPH assay highlighted that eight of the samples (Lesvos and Lemnos origin) demonstrated a promising antioxidant profile, further verified by their total phenolic content (TPC). Additionally, the propolis samples most rich in diterpenes showed significant antibacterial and fungicidal properties against human pathogenic microorganisms, proving them to be a very interesting and promising crude material for further applications, concluding that floral diversity is the most responsible for the bioactivity of the propolis samples.

Conformational Properties and Putative Bioactive Targets for Novel Thiosemicarbazone Derivatives.

The structure assignment and conformational analysis of the thiosemicarbazones, DKI21 and DKI24, were performed through homonuclear and heteronuclear 2D Nuclear Magnetic Resonance (NMR) spectroscopy (2D-COSY, 2D-NOESY, 2D-ROESY, 2D-HSQC, and 2D-HMBC) and quantum mechanics (QM) calculations, using Functional Density Theory (DFT). In addition, utilizing a combination of 2D-NOESY and 2D-ROESY spectra an exo structure was established for both of the analogs. This experimental results were confirmed by theoretical mechanistic studies, as the lowest minima conformations derived through DFT calculations were compatible with the spatial correlations observed in the 2D-NOESY and 2D-ROESY spectra. Finally, molecular binding experiments were performed to detect the potential targets for DKI21 and DKI24, derived from SwissAdme. In silico molecular binding experiments showed favorable binding energy values for the most of the enzymes studied. The ADMET calculations, using the preADMET and pKCSm software, showed that the two molecules appear as possible drug leads.

Untargeted UHPLC-MS metabolic profiling as a valuable tool for the evaluation of eggs quality parameters after dietary supplementation with oregano, thyme, sideritis tea and chamomile on brown laying hens.

INTRODUCTION: Bioactive constituents of medicinal-aromatic plants used as feed additives may affect the metabolic profile and oxidative stability of hen eggs. OBJECTIVES: To determine the effects of dietary supplementation with a mixture of dried oregano, thyme, sideritis tea and chamomile on laying hen performance, egg quality parameters, and oxidative stability in the egg yolk were monitored. METHODS: In this trial 432 hens were allocated in two treatments (unsupplemented vs. supplemented with the mixture) and fed for 42 days. Eggs were collected at the end of the trial period, egg yolk was separated, extracted, and the total phenolic content (TPC) and oxidative stability was measured. Furthermore, LC-MS metabolic profile of eggs was studied and pathway analysis was elaborated in MetaboAnalyst to facilitate annotation of features. RESULTS: Overall, egg production and feed conversion ratio were not affected by the supplementation. However, eggs from the supplemented treatment showed improved shell thickness and strength, and yolk resistance to oxidation. Moreover, LC-MS metabolomic analysis of egg yolk of supplemented and unsupplemented layers showed significant variations and tight clustering in unsupervised principal component analysis due to different chemical profiling of egg yolk. LC-MS study showed that secondary metabolites of aromatic plants did not transfer into yolk, nevertheless the feed supplementation impacted the pathway metabolism of tyrosine, phenylalanine, propanate, and the biosynthesis of aminoacyl-tRNA, phenylalanine, tyrosine and tryptophan. CONCLUSIONS: The dietary supplementation of layers with a mixture of dried medicinal aromatic plants affected shell thickness and strength, the lipid and protein oxidative stability and increased tyrosine and phenylalanine content in eggs.

Untargeted Ultrahigh-Performance Liquid Chromatography-Hybrid Quadrupole-Orbitrap Mass Spectrometry (UHPLC-HRMS) Metabolomics Reveals Propolis Markers of Greek and Chinese Origin.

Chemical composition of propolis depends on the plant source and thus on the geographic and climatic characteristics of the site of collection. The aim of this study was to investigate the chemical profile of Greek and Chinese propolis extracts from different regions and suggest similarities and differences between them. Untargeted ultrahigh-performance liquid chromatography coupled to hybrid quadrupole-Orbitrap mass spectrometry (UHPLC-HRMS) method was developed and 22 and 23 propolis samples from Greece and China, respectively, were analyzed. The experimental data led to the observation that there is considerable variability in terms of quality of the distinctive propolis samples. Partial least squares - discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) models were constructed and allowed the identification of significant features for sample discrimination, adding relevant information for the identification of class-determining metabolites. Chinese samples overexpressed compounds that are characteristic of the poplar type propolis, whereas Greek samples overexpress the latter and the diterpenes characteristic of the Mediterranean propolis type.

Isoflavonoid Profiling and Estrogen-Like Activity of Four Genista Species from the Greek Flora.

As part of our ongoing research on phytoestrogens, we investigated the phytochemical profile and estrogen-like activities of eight extracts from the aerial parts of four Genista species of Greek flora using estrogen-responsive cell lines. Ethyl acetate and methanolic extracts of G. acanthoclada, G. depressa,G. hassertiana, and G. millii were obtained with accelerated solvent extraction and their phytochemical profiles were compared using ultra-high-performance liquid chromatography-high-resolution mass spectrometry (uHPLC-HRMS). Fourteen isoflavonoids, previously isolated from G. halacsyi, were used as reference standards for their identification in the extracts. Thirteen isoflavonoids were detected in both extracts of G. acanthoclada and G. hassertiana, while fewer and far fewer were detected in G. millii and G. depressa, respectively. The ethyl acetate extracts of G. hassertiana and G. acanthoclada displayed 2.45- and 1.79-fold higher, respectively, estrogen-like agonist activity in Ishikawa cells compared to MCF-7 cells at pharmacologically relevant concentrations. Both these extracts, but not that of G. depressa, contained mono- and di-O-ß-d-glucosides of genistein as well as the aglycone, all three of which are known to display full estrogen-like activity at lower-than-micromolar concentrations. The possibility of using preparations rich in G. hassertiana and/or G. acanthoclada extracts as a potentially safer substitute for low-dose vaginal estrogen for menopausal symptoms is discussed.

Oregano and thyme by-products of olive oil aromatization process with microwave assisted extraction as a rich source of bio-active constituents.

Introduction: Processing of Medicinal Aromatic Plants (MAPs) results in the production of a significant amount of by-products, which are not commercially exploitable. Towards this direction, we studied extensively the by-products of oregano and thyme, remaining after the aromatization of olive oils with microwave assisted extraction (MAE). The purpose of the study was the exploitation of the "wastes" of these two economically significant herbs of Greece, for the potential development of innovative bioactive products. Methods: Hence, superior and inferior quality plant material from Origanum vulgare subsp. hirtum and Thymus vulgaris, were extracted with extra virgin olive oil using MAE. For the evaluation of raw plant material, beside the characterization of the essential oils (EOs), the hydroalcoholic extracts of superior and inferior plant material were afforded by ultrasound assistant extraction (UAE). In addition, the remaining plant material after the flavoring of olive oil by MAE, was extracted with c-Hex, MeOH, H2O:MeOH using UAE. All the extracts were evaluated for their DPPH free radical scavenging activity and total phenolic content (TPC) as well as their chemical profile was investigated by HPTLC. In parallel, the EOs, the olive oils and the c-Hex extracts were analyzed by GC-MS and Headspace Solid Phase Microextraction (HS-SPME)-GC-MS. Results and Discussion: The results showed that the composition of the EOs and the volatile fraction of the olive oil extracts were similar for the superior quality material whereas for the inferior the composition of the volatile fraction of olive oil extracts was not analogous to the respective EOs. GC-MS analyses of oregano and thyme by-products revealed the presence of carvacrol, thymol, γ-terpinene and p-cymene among the major constituents. Moreover, the hydroalcoholic extracts obtained from the plant material remaining after olive oil flavoring with MAE showed similar phenolic content and scavenging activity with the hydroalcoholic extracts of the corresponding raw plant materials underlying their potent use in the preparation of high-added value products such as nutraceuticals and cosmeceuticals as well as enriched animal nutrition products.

Ethnopharmacological study of medicinal plants used against skin ailments on Mount Pelion, central Greece.

Throughout centuries, traditional herbal medicine and the employment of medicinal plants have constituted an important tool for the treatment and prevention of numerous diseases. The present study focuses on the collection of ethnopharmacological data regarding the uses of medicinal plants for the treatment of dermatological ailments in various villages of Mount Pelion, Greece. More specifically, the study area is represented by the city of Volos and villages located in Central West Pelion and has not been investigated up to now. The information on the medicinal uses of the various species was obtained through extensive semi-structured interviews or the completion of specific questionnaires by the informants. Although the Covid-19 pandemic caused difficulties and obstacles in carrying out this research procedure, 60 informants were recruited and interviewed (36 women and 24 men). Their age range was between 31 and 97 years and their educational level was characterized by great diversity (primary, secondary, and higher education). The elaboration of the gathered information included the calculation of some quantitative indices, such as Fidelity Level (FL), and Informant Consensus Factor (FIC). Moreover, the relative importance of each reported species was identified by calculating the Use Value (UV). The interviews revealed 38 plant taxa belonging to 27 plant families reported to be used in the study area exclusively against skin diseases. The plant family mostly mentioned by the informants was Hypericaceae, followed by Plantaginaceae and Amaryllidaceae, while among the most popular methods of application are cataplasms, compresses, and topical application of decoction or raw plant material. Some of the most cited species are Hypericum perforatum L., Quercus coccifera L., and Plantago sp., traditionally used to treat skin problems such as eczema, wounds, and insect stings. The present ethnopharmacological study is the first documentation of ethnobotanical knowledge of this area that points out the traditional uses of medicinal plants against skin ailments.

Metabolomic Profiling of Second-Trimester Amniotic Fluid for Predicting Preterm Delivery: Insights from NMR Analysis.

Preterm delivery (PTD) is a notable pregnancy complication, affecting one out of every ten births. This study set out to investigate whether analyzing the metabolic composition of amniotic fluid (AF) collected from pregnant women during the second trimester of pregnancy could offer valuable insights into prematurity. The research employed 1H-NMR metabolomics to examine AF samples obtained from 17 women who gave birth prematurely (between 29+0 and 36+5 weeks of gestation) and 43 women who delivered at full term. The application of multivariate analysis revealed metabolites (dimethylglycine, glucose, myo-inositol, and succinate) that can serve as possible biomarkers for the prognosis and early diagnosis of preterm delivery. Additionally, pathway analysis unveiled the most critical metabolic pathways relevant to our research hypothesis. In summary, these findings suggest that the metabolic composition of AF in the second trimester can be a potential indicator for identifying biomarkers associated with the risk of PTD.

Thiocarbohydrazone and Chalcone-Derived 3,4-Dihydropyrimidinethione as Lipid Peroxidation and Soybean Lipoxygenase Inhibitors.

The potential of the 4,6-diphenyl-3,4-dihydropyrimidine-2(1H)-thione (abbreviated as KKII5) and (E)-N'-benzylidenehydrazinecarbothiohydrazide (abbreviated as DKI5) compounds as possible drug leads is investigated. KKII5 and DKI5 are synthesized in high yield of up to 97%. Their structure, binding in the active site of the LOX-1 enzyme, and their toxicity are studied via joint experimental and computational methodologies. Specifically, the structure assignment and conformational analysis were achieved by applying homonuclear and heteronuclear 2D nuclear magnetic resonance (NMR) spectroscopy (2D-COSY, 2D-NOESY, 2D-HSQC, and 2D-HMBC) and density functional theory (DFT). The obtained DFT lowest energy conformers were in agreement with the NOE correlations observed in the 2D-NOESY spectra. Additionally, docking and molecular dynamics simulations were performed to discover their ability to bind and remain stabile in the active site of the LOX-1 enzyme. These in silico experiments and DFT calculations indicated favorable binding for the enzyme under study. The strongest binding energy, -9.60 kcal/mol, was observed for dihydropyrimidinethione KKII5 in the active site of LOX-1. ADMET calculations showed that the two molecules lack major toxicities and could serve as possible drug leads. The redox potential of the active center of LOX-1 with the binding molecules was calculated via DFT methodology. The results showed a significantly smaller energy attachment of 2.8 eV with KKII5 binding in comparison to DKI5. Thus, KKII5 enhanced the ability of the active center to receive electrons compared to DKI5. This is related to the stronger binding interaction of KKII5 relative to that of DK15 to LOX-1. The two very potent LOX-1 inhibitors exerted IC50 19 µΜ (KKII5) and 22.5 µΜ (DKI5). Furthermore, they both strongly inhibit lipid peroxidation, namely, 98% for KKII5 and 94% for DKI5.

Sustainable Use of Greek Herbs By-Products, as an Alternative Source of Biologically Active Ingredients for Innovative Products.

The processing of medicinal and aromatic plants (MAPs) results in the production of a significant amount of plant by-products; herbal material of inferior quality and/or unusable plant parts that are not commercially exploitable. An extensive study of Greek native species was performed toward the production of innovative bioactive products using as raw materials the by-products obtained from the processing of cultivated MAPs. Origanum vulgare subsp. hirtum (oregano), Sideritis scardica (Greek mountain tea), Thymus vulgaris (thyme), and Matricaria recutita (chamomile) were selected due to their wide use for the preparation of beverages and culinary purposes. The determination of the percentage of the post-harvest processing by-products was performed for a 3 years period (2018-2020). Results showed that by-products derived from the above-mentioned species' processing constitute 64% (thyme), 54% (oregano), 37% (Greek mountain tea), and 24% (chamomile) of the total processed mass. To value the by-products as a potent source of bioactive ingredients, superior and inferior quality herbal material of the aforementioned plant species were extracted by an ultrasonic assisted extraction method. Hydroalcoholic extracts were chemically investigated using high-performance thin layer chromatography (HPTLC) and liquid chromatography-mass spectrometry (LC-MS) techniques. In addition, their free radical scavenging activity and total phenolic content (TPC) were estimated. Based on the results, herbs by-products revealed similar chemical content to the superior herbal material by the means of HPTLC and LC-MS analysis. In addition, strong free radical scavenging related to a high phenolic content was detected in the case of thyme, oregano, and Greek mountain tea. Moreover, the gas chromatography-mass spectrometry (GC-MS) analyses of the essential oils (EOs) of oregano and thyme by-products revealed the presence of carvacrol, thymol, γ-terpinene, and p-cymene among the major constituents. Finally, the LC-MS analyses of aqueous extracts of Greek mountain tea and chamomile by-products led to the identification of several bioactive compounds, such as flavonoids and phenylpropanoids. Overall, the presence of bioactive constituents in by-products, such as terpenes, phenolic compounds, and flavonoids underly their potent use as food antimicrobial and antioxidant additives, in the preparation of high added-value products, such as enriched aromatic edible oils, and innovative herbal teas, such as instant beverages.

Medicinal Plants Used Traditionally for Skin Related Problems in the South Balkan and East Mediterranean Region-A Review.

A review research was conducted to provide an overview of the ethnobotanical knowledge of medicinal plants and traditional medical practices for the treatment of skin disorders in Albania, Cyprus, Greece, and Turkey. The geographical and ecological characteristics of the Balkan Peninsula and Mediterranean Sea, along with the historical connection among those countries, gave rise to the development of a distinct flora and to the uses of common medicinal plants against various skin ailments, respectively. The review focuses on the detailed study of 128 ethnobotanical surveys conducted in these areas and the species used for skin ailments were singled out. The analysis showed that 967 taxa belonging to 418 different genera and 111 different families are used in the treatment of skin related problems. The majority of the plants belong to the families of Asteraceae (11.7%), Lamiaceae (7.4%), Rosaceae (6.7%), Plantaginaceae (5.4%), and Malvaceae (3.8%). Their usage is internal or external to treat ailments such as wounds and burns (22.1%), hemorrhoids (14.7%), boils, abscesses, and furuncles (8.2%). Beside specific skin disorders, numerous species appeared to be used for their antifungal, antimicrobial, and antiseptic activity (9.1%). Literature evaluation highlighted that, the most commonly used species are Plantago major L. (Albania, Turkey), Hypericum perforatum L. (Greece, Turkey), Sambucus nigra L. (Cyprus, Greece), Ficus carica L. (Cyprus, Turkey), Matricaria chamomilla L. (Cyprus, Greece), and Urtica dioica L. (Albania, Turkey), while many medicinal plants reported by interviewees were common in all four countries. Finally, to relate this ethnopharmacological knowledge and trace its expansion and diversification through centuries, a comparison of findings was made with the use of the species mentioned in Dioscorides' "De Materia Medica" for skin disorders. This work constitutes the first comparative study performed with ethnobotanical data for skin ailments gathered in the South Balkan and East Mediterranean areas. Results confirm the primary hypothesis that people in Albania, Cyprus, Greece, and Turkey are closely related in terms of traditionally using folk medicinal practices. Nevertheless, more field studies conducted, especially in remote places of these regions, can help preserve the traditional medical knowledge, aiming at the discovery of new phytotherapeutics against dermatological diseases.

Chemical and Biological Evaluation of the Oil and Seedcake from Seeds of a Greek Cardoon Cultivar as Potential Functional Vegetable Oil. Comparison with Sesame, Flaxseed and Extra Virgin Olive Oils.

Cynara cardunculus L. is a plant of the Mediterranean basin, known since antiquity as a food and for its therapeutic properties. The needs of the 21st century for the utilization of agricultural waste has led to the study of the seed oil of a Greek cultivar of Cynaracardunculus (GCCC) as potential nutritional oil, as large amounts of cardoon seeds are discarded. The sterol and fatty acid profile of cold-pressed seed oil was examined by gas chromatography-mass spectrometry GC-MS and compared with that of solvent extraction. Total phenolic content was determined and compared with well-known and widely appreciated edible vegetable oils; while, additionally, the total lignan content and nutritional value of cold-pressed oil revealed it as a potential dietary candidate. Furthermore, the seedcake (residue of cold-pressed oil extraction) has been studied exerting it as a good source of phenolics. Both GCCC oil and seedcake were tested for their antioxidant and enzyme inhibitory activities exhibiting higher activity compared to the sesame, flaxseed and extra virgin olive oils. According to the results, Cynara seed oil was shown to be a rich source of ω-6/-9 fatty acids and phenolics, highlighting, indicating that it could be a promising health-promoting vegetable oil, while the seedcake was revealed as a rich source of bioactive compounds.

NMR metabolic profiling of Greek propolis samples: Comparative evaluation of their phytochemical compositions and investigation of their anti-ageing and antioxidant properties.

The present study aimed to investigate the metabolic profile, as well as the antioxidant and anti-ageing activities of twenty propolis samples from different regions of Greece. Chemical profiling of methanolic extracts was investigated using HPTLC and 1H-NMR techniques. Their antioxidant activity was evaluated by free radical scavenging methods (DPPH and ABTS), whereas anti-ageing properties were assessed as anti-collagenase activity. Extracts were also investigated in vitro for their ability to inhibit tyrosinase, which is responsible for the oxidation of L-DOPA to dopachrome and the production of melanin. The HPTLC and NMR analysis revealed high variability in the phytochemical profile of the methanolic extracts, with three major groups to be observed: a) Group I, consisting of samples rich in terpenoids, which present low antioxidant but high anti-tyrosinase activity, b) Group II, consisting of samples rich in flavonoids, which form a broad cluster with major similarities at the aromatic region and showed the highest anti-oxidant and anti-collagenase activities and c) Group III, consisting of samples with lower flavonoid content than the samples of Group II, which exhibited moderate antioxidant, anti-collagenase and anti-tyrosinase activities. In conclusion, this study has shown high differentiation on the chromatographic and spectroscopic metabolic profile of Greek propolis samples of different geographical origin, which is also reflected in their biological properties. Their important effects as antioxidant, anti-tyrosinase and anti-collagenase agents make propolis an important potent ingredient in the industry of food supplements and cosmeceuticals. Moreover, a correlation of a particular chemical propolis type to a specific type of biological activity will allow to prepare standardized extracts and develop food supplements and cosmeceuticals possessing the desired pharmacological properties.

An enriched polyphenolic extract obtained from the by-product of Rosa damascena hydrodistillation activates antioxidant and proteostatic modules.

BACKGROUND: Prolonged maintenance of proteome stability and functionality (proteostasis) is of emerging significance in aging retardation and healthspan. PURPOSE: An enriched polyphenolic extract obtained from the hydrodistillation of rose petals was tested for its capacity to activate the proteostasis network modules, and thus modulate health- and/or lifespan at the cellular and whole organism level. METHODS: The aqueous extract that remained after the hydrodistillation of Rosa damascena petals, was processed with a polystyrene-FPX66 adsorption resin and sequentially fractionated by FCPC. NMR and UHPLC-HRMS analyses revealed the presence of 28 metabolites, mainly glycosides of kaempferol and quercetin. RESULTS: The extract showed high in vitro antioxidant activity and was not toxic in normal human skin fibroblasts, while it promoted the upregulation of NRF2-induced antioxidant genes and main proteostatic modules. Consistently, supplementation of this extract in Drosophila flies' culture medium induced a cncC/NRF2-mediated upregulation of antioxidant and proteostatic modules. Prolonged administration of the extract in flies' culture medium was not toxic and did not affect food intake rate or fecundity; also, it delayed the age-related decline of stress tolerance and locomotion performance (neuromuscular functionality) and dose-dependently extended flies' lifespan. CONCLUSION: Our findings indicate that the enriched polyphenolic extract obtained from the residue of R. damascena hydrodistillation activates cytoprotective cellular modules that, likely, contribute to its potential anti-aging properties.

Biological evaluation of isoflavonoids from Genista halacsyi using estrogen-target cells: Activities of glucosides compared to aglycones.

The purpose of this study was to evaluate the response of estrogen target cells to a series of isoflavone glucosides and aglycones from Genista halacsyi Heldr. The methanolic extract of aerial parts of this plant was processed using fast centrifugal partition chromatography, resulting in isolation of four archetypal isoflavones (genistein, daidzein, isoprunetin, 8-C-ß-D-glucopyranosyl-genistein) and ten derivatives thereof. 7-O-ß-D-glucopyranosyl-genistein and 7,4΄-di-O-ß-D-glucopyranosyl-genistein were among the most abundant constituents of the isolate. All fourteen, except genistein, displayed low binding affinity for estrogen receptors (ER). Models of binding to ERα could account for the low binding affinity of monoglucosides. Genistein and its glucosides displayed full efficacy in inducing alkaline phosphatase (AlkP) in Ishikawa cells, proliferation of MCF-7 cells and ER-dependent gene expression in reporter cells at low concentrations (around 0.3 µM). ICI182,780 fully antagonized these effects. The AlkP-inducing efficacy of the fourteen isoflavonoids was more strongly correlated with their transcriptional efficacy through ERα. O-monoglucosides displayed higher area under the dose-response curve (AUC) of AlkP response relative to the AUC of ERα-transcriptional response compared to the respective aglycones. In addition, 7-O-ß-D-glucopyranosyl-genistein and 7,4΄-di-O-ß-D-glucopyranosyl-genistein displayed estradiol-like efficacy in promoting differentiation of MC3T3-E1 cells to osteoblasts, while genistein was not convincingly effective in this respect. Moreover, 7,4΄-di-O-ß-D-glucopyranosyl-genistein suppressed lipopolysaccharide-induced tumor necrosis factor mRNA expression in RAW 264.7 cells, while 7-O-ß-D-glucopyranosyl-genistein was not convincingly effective and genistein was ineffective. However, genistein and its O-glucosides were ineffective in inhibiting differentiation of RAW 264.7 cells to osteoclasts and in protecting glutamate-challenged HT22 hippocampal neurons from oxidative stress-induced cell death. These findings suggest that 7-O-ß-D-glucopyranosyl-genistein and 7,4΄-di-O-ß-D-glucopyranosyl-genistein display higher estrogen-like and/or anti-inflammatory activity compared to the aglycone. The possibility of using preparations rich in O-ß-D-glucopyranosides of genistein to substitute for low-dose estrogen in formulations for menopausal symptoms is discussed.

Head-to-Head Comparison of Ultra-High-Performance Liquid Chromatography with Diode Array Detection versus Quantitative Nuclear Magnetic Resonance for the Quantitative Analysis of the Silymarin Complex in Silybum marianum Fruit Extracts.

Quantitative nuclear magnetic resonance (qNMR) spectroscopy is known as an excellent alternative to chromatography-based mixture analysis. NMR spectroscopy is a non-destructive method, needs only limited sample preparation, and can be readily automated. A head-to-head comparison of qNMR to an ultra-high-performance liquid chromatography with diode array detection (uHPLC-DAD)-based quantitative analysis of six flavonolignan congeners (silychristin, silydianin, silybin A, silybin B, isosilybin A, and isosilybin B) of the Silybum marianum silymarin complex is presented. Both assays showed similar performance characteristics (linear range, accuracy, precision, and limits of quantitation) with analysis times below 30 min/sample. The assays were applied to industrial S. marianum extracts (AC samples) and to extracts locally prepared from S. marianum fruits (PL samples). An assay comparison by Bland-Altman plots (relative method bias AC samples, -0.1%; 2SD range, ±5.1%; relative method bias PL samples, -0.3%; 2SD range, ±7.8%) and Passing-Bablok regression analysis (slope and intercept for AC and PL samples not significantly different from 1.00 and 0.00, respectively; Spearman's coefficient of rank correlation, >0.99) did show that qNMR and uHPLC-DAD can be used interchangeably to quantitate flavonolignans in the silymarin complex.