Eleftherios Kalpoutzakis

Characteristic manifestations of skin aging, due to either intrinsic or extrinsic factors, such as ultraviolet (UV) radiation and oxidative stress, include cell senescence, alterations in collagen and elastin networks, and melanogenesis disorders. Natural products are considered a rich source of anti-aging molecules. Accordingly, the screening of a plant extract library from the Greek flora for a panel of biological activities related to skin aging is presented herein. In particular, 52 plant materials extracted using Accelerated Solvent Extraction (ASE) and Supercritical Fluid Extraction (SFE) were assessed for their effects on (1) human skin cell viability, (2) antioxidant activity—using both cell-free and cell-based methods—(3) photoprotective capacity, and (4) interference with collagenase, elastase, and tyrosinase, as well as with proteasomal and lysosomal activities of human skin cells. In vivo phenotypic screens on Danio rerio (zebrafish) embryos were also used for assessing melanogenesis. Many active extracts were identified, some of them for the first time, and others in agreement with previous reports. In general, ASE extracts exhibited higher activities than SFE ones. Seven extracts showed multiple activities, being highly effective in at least four different assays. These data support the potential use of these extracts against skin aging in medicinal and cosmetic applications.

The abundant yet underutilized olive leaves, a renewable by-product of olive cultivation, offer untapped potential for producing high-value bioactive compounds, notably oleacein. Existing extraction methods are often inefficient, yielding low quantities of oleacein due to enzymatic degradation of its precursor, oleuropein, during conventional processing and storage. This study aimed to overcome these limitations by exploring a novel methodology based on freeze-drying, to facilitate the in situ enzymatic biotransformation of oleuropein into oleacein directly within the plant matrix. Olive leaves were subjected to three drying methods—ambient air drying, microwave drying, and freeze-drying—and their phenolic profiles were analyzed. The findings demonstrated that freeze drying uniquely promotes the selective activation of β-glucosidase and esterase enzymes while simultaneously inhibiting oxidative enzymes, such as polyphenol oxidase and peroxidase, resulting in significantly enriched oleacein content. This process eliminates the need for extensive post-extraction transformations, providing a cost-effective, scalable, and sustainable approach to oleacein production. The proposed methodology aligns with circular economy principles and holds substantial potential for applications in pharmaceuticals, nutraceuticals, and functional food industries.

Oxidative damages are responsible for many adverse health effects and food deterioration. The use of antioxidant substances is well renowned, and as such, much emphasis is placed on their use. Since synthetic antioxidants exhibit potential adverse effects, plant-derived antioxidants are a preferable solution. Despite the myriads of plants that exist and the fact that numerous studies have been carried out so far, there are many species that have not been examined so far. Many plants under research exist in Greece. Trying to fill this research gap, the total phenolics content and antioxidant activity of seventy methanolic extracts from parts of Greek plants were evaluated. The total phenolics content was measured by the Folin–Ciocalteau assay. Their antioxidant capacity was calculated by the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging test, the Rancimat method based on conductometric measurements, and the thermoanalytical method DSC (Differential Scanning Calorimetry). The tested samples were obtained from several parts of fifty-seven Greek plant species belonging to twenty-three different families. Both a remarkably high phenolic content (with gallic acid equivalents varying between 311.6 and 735.5 mg/g of extract) and radical scavenging activity (IC₅₀ values ranged from 7.2 to 39.0 μg/mL) were found in the extract of the aerial parts of Cistus species (C. creticus subsp. creticus, C. creticus subsp. eriocephalus, C. monspeliensis, C. parviflorus and C. salviifolius), Cytinus taxa (C. hypocistis subsp. hypocistis, C. hypocistis subsp. orientalis and C. ruber), and Sarcopoterium spinosum. Furthermore, the sample of Cytinus ruber showed the highest protection factor (PF = 1.276) regarding the Rancimat method, which was similar to that of butylated hydroxytoluene (BHT) (PF = 1.320). The results indicated that these plants are rich in antioxidant compounds, potentiating their use either as food additives to enhance the antioxidant properties of food products, or protect them from oxidation, or as sources for the preparation of food supplements with antioxidant properties.

Introduction: The genus Origanum L. (Lamiaceae) is widespread in the Mediterranean region. However, approximately 75% of the species are only encountered in the eastern part. Out of these, a total of nine species (11 taxa) and three natural hybrids occur in Greece. Nevertheless, so far, there is no consensus regarding their precise botanical classification in the literature. In fact, the taxon Origanum × lirium has been proposed both as a separate species as well as natural hybrid between Origanum vulgare subsp. hirtum and Origanum scabrum.Objectives: In this scope, the aim of the current study is to shed light on the matter through the investigation of the chemical composition of both the essential oils and the polar extracts of the mentioned taxa, collected from different geographical regions of Greece.Results: As it was demonstrated by both gas chromatography–mass spectrometry (GC‐MS) and liquid chromatography–high‐resolution mass spectrometry (LC–HRMS) data, and highlighted by our comparative analysis, it can be stipulated that Origanum × lirium shares its chemotype to a large extent with its parent species concerning both volatile and polar constituents. Additionally, geographical origin conditions stood out as a key factor influencing their chemical composition.Conclusion: Altogether, the present work provides useful information on the chemical composition of the taxa under investigation, while our findings support the opinion that Origanum × lirium should be considered not as a separate species, but rather as a hybrid on the way to becoming a species.

This study was designed to evaluate the chemical fingerprints and the antioxidant, anti-inflammatory and hypolipidemic activity of selected Crepis species collected in Greece, namely, C. commutata, C. dioscoridis, C. foetida, C. heldreichiana, C. incana, C. rubra, and Phitosia crocifolia (formerly known as Crepis crocifolia). For the phytochemical analyses, sample measurements were carried out by using nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography coupled with mass spectrometry (LC-MS). Τhe extracts were evaluated both in vitro (radical scavenging activity: DPPH assay and total phenolic content: Folin–Ciocalteu) and in vivo (paw edema reduction and hypolipidemic activity: experimental mouse protocols). Among the tested extracts, C. incana presented the highest gallic acid equivalents (GAE) (0.0834 mg/mL) and the highest antioxidant activity (IC₅₀ = 0.07 mg/mL) in vitro, as well as the highest anti-inflammatory activity with 32% edema reduction in vivo. Moreover, in the hypolipidemic protocol, the same extract increased plasma total antioxidant capacity (TAC) by 48.7%, and decreased cholesterol (41.3%) as well as triglycerides (37.2%). According to fractionation of the extract and the phytochemical results, this biological effect may be associated with the rich phenolic composition; caffeoyl tartaric acid derivatives (cichoric and caftaric acid) are regarded as the most prominent bioactive specialized metabolites. The present study contributes to the knowledge regarding the phytochemical and pharmacological profile of Crepis spp.

Palmatine (PLT) is a natural isoquinoline alkaloid that belongs to the class of protoberberines and exhibits a wide spectrum of pharmacological and biological properties, including anti-cancer activity. The aim of our study was to isolate PLT from the roots of Berberis cretica and investigate its cytotoxic and anti-proliferative effects in vitro alone and in combination with doxorubicine (DOX) using human ER⁺/HER2⁻ breast cancer cell lines. The alkaloid was purified by column chromatography filled with silica gel NP and Sephadex LH-20 resin developed in the mixture of methanol: water (50:50 v/v) that provided high-purity alkaloid for bioactivity studies. The purity of the alkaloid was confirmed by high resolution mass measurement and MS/MS fragmentation analysis in the HPLC-ESI-QTOF-MS/MS-based analysis. It was found that PLT treatment inhibited the viability and proliferation of breast cancer cells in a dose-dependent manner as demonstrated by MTT and BrdU assays. PLT showed a quite similar growth inhibition on breast cancer cells with IC₅₀ values ranging from 5.126 to 5.805 µg/mL. In contrast, growth of normal human breast epithelial cells was not affected by PLT. The growth inhibitory activity of PLT was related to the induction of apoptosis, as determined by Annexin V/PI staining. Moreover, PLT sensitized breast cancer cells to DOX. Isobolographic analysis revealed synergistic and additive interactions between studied agents. Our studies suggest that PLT can be a potential candidate agent for preventing and treating breast cancer.

Skin health is heavily affected by ultraviolet irradiation from the sun. In addition, senile skin is characterized by major changes in the collagen, elastin and in the hyaluronan content. Natural products (NPs) have been shown to delay cellular senescence or in vivo aging by regulating age-related signaling pathways. Moreover, NPs are a preferable source of photoprotective agents and have been proven to be useful against the undesirable skin hyperpigmentation. Greek flora harvests great plant diversity with approximately 6000 plant species, as it has a wealth of NPs. Here, we report an extensive screening among hundreds of plant species. More than 440 plant species and subspecies were selected and evaluated. The extracts were screened for their antioxidant and anti-melanogenic properties, while the most promising were further subjected to various in vitro and cell-based assays related to skin aging. In parallel, their chemical profile was analyzed with High-Performance Thin-Layer Chromatography (HPTLC) and/or Ultra-Performance Liquid Chromatography High-Resolution Mass Spectrometry (UPLC-HRMS). A variety of extracts were identified that can be of great value for the cosmetic industry, since they combine antioxidant, photoprotective, anti-melanogenic and anti-aging properties. In particular, the methanolic extracts of Sideritis scardica and Rosa damascena could be worthy of further attention, since they showed interesting chemical profiles and promising properties against specific targets involved in skin aging.

Ethnopharmacology experts face several challenges when identifying and retrieving documents and resources related to their scientific focus. The volume of sources that need to be monitored, the variety of formats utilized, the different quality of language use across sources, present some of what we call “big data” challenges in the analysis of this data. This study aims to understand if and how experts can be supported effectively through intelligent tools in the task of ethnopharmacological literature research. To this end, we utilize a real case study of ethnopharmacology research, aimed at the Southern Balkans and Coastal zone of Asia Minor. Thus, we propose a methodology for more efficient research in ethnopharmacology. Our work follows an “Expert-Apprentice” paradigm in an automatic URL extraction process, through crawling, where the apprentice is a Machine Learning (ML) algorithm, utilizing a combination of Active Learning (AL) and Reinforcement Learning (RL), and the Expert is the human researcher. ML-powered research improved 3.1 times the effectiveness and 5.14 times the efficiency of the domain expert, fetching a total number of 420 relevant ethnopharmacological documents in only 7 hours versus an estimated 36-hour human-expert effort. Therefore, utilizing Artificial Intelligence (AI) tools to support the researcher can boost the efficiency and effectiveness of the identification and retrieval of appropriate documents.

Ethnopharmacology experts face several challenges when identifying and retrieving documents and resources related to their scientific focus. The volume of sources that need to be monitored, the variety of formats utilized, and the different quality of language use across sources present some of what we call “big data” challenges in the analysis of this data. This study aims to understand if and how experts can be supported effectively through intelligent tools in the task of ethnopharmacological literature research. To this end, we utilize a real case study of ethnopharmacology research aimed at the southern Balkans and the coastal zone of Asia Minor. Thus, we propose a methodology for more efficient research in ethnopharmacology. Our work follows an “expert–apprentice” paradigm in an automatic URL extraction process, through crawling, where the apprentice is a machine learning (ML) algorithm, utilizing a combination of active learning (AL) and reinforcement learning (RL), and the expert is the human researcher. ML-powered research improved the effectiveness and efficiency of the domain expert by 3.1 and 5.14 times, respectively, fetching a total number of 420 relevant ethnopharmacological documents in only 7 h versus an estimated 36 h of human-expert effort. Therefore, utilizing artificial intelligence (AI) tools to support the researcher can boost the efficiency and effectiveness of the identification and retrieval of appropriate documents.