Enrichment of Olive Oils with Natural Bioactive Compounds from Aromatic and Medicinal Herbs: Phytochemical Analysis and Antioxidant Potential.

Olive oil and herbs, two key components of the Mediterranean diet, are known for their beneficial effects on humans. In our study, we incorporated aromatic and medicinal herbs into local monovarietal olive oils via maceration procedures for enrichment. We identified the herbal-derived ingredients that migrate to olive oils and contribute positively to their total phenolic content and functional properties, such as radical scavenging activity. Thus, we characterized the essential oil composition of the aromatic herbs (GC-MS), and we determined the phenolic content and antioxidant capacity of the additives and the virgin olive oils before and after enrichment. The herbal phenolic compounds were analyzed by LC-LTQ/Orbitrap HRMS. We found that olive oils infused with Origanum vulgare ssp. hirtum, Rosmarinus officinalis and Salvia triloba obtained an increased phenolic content, by approximately 1.3 to 3.4 times, in comparison with the untreated ones. Infusion with S. triloba led to a significantly higher antioxidant capacity. Rosmarinic acid, as well as phenolic glucosides, identified in the aromatic herbs, were not incorporated into olive oils due to their high polarity. In contrast, phenolic aglycones and diterpenes from R. officinalis and S. triloba migrated to the enriched olive oils, leading to a significant increase in their phenolic content and to an improvement in their free radical scavenging capacity.

A Multifaceted Evaluation Approach for Greek Native Neglected and Underutilized Forest Fruit Trees and Shrubs as Natural Sources of Antioxidants: Consolidating the Framework for Their Sustainable Agronomic Exploitation.

Fruits from wild forest trees and shrubs represent a natural source of antioxidants against oxidative stress and a growing market for novel minor crops. This study presents a multifaceted approach which sets the basis for sustainable agronomic exploitation of selected Greek native germplasm of four traditional but neglected and underutilized forest fruit trees and shrubs, namely Amelanchier ovalis Medik., Cornus mas L., Rosa canina L., and Sambucus nigra L. The studied plant species are traditionally used in Greek ethnobotany but are currently neglected and underutilized in commercial terms, thus characterized as neglected and underutilized plant species (NUPs). The investigation includes new information on the evaluation of the ex situ cultivation of the Greek germplasm (three of the four focal NUPs), thus supplementing respective full datasets for their comparative evaluation based on four evaluation axes (documentation and molecular authentication of genotypes, phytochemical evaluation, asexual propagation via rooting of cuttings, and ex situ cultivation) after multi-year and multifaceted groundwork data previously acquired. Consecutively, the work includes feasibility and readiness timescale evaluation for the sustainable exploitation of each focal species based on existing literature and extant research experience. The feasibility for sustainable exploitation and readiness timescale evaluation results were very encouraging, showing high exploitation feasibility with an already achieved readiness timescale for R. canina and S. nigra, whereas C. mas and A. ovalis showed an achievable readiness in the short term. The comparative evaluation of the Greek native focal NUPs outlined the excellent potential of R. canina, S. nigra, and A. ovalis, and the high potential of C. mas. The results herein illustrate the very high fruit antioxidant potential (free radical scavenging activity) of all focal species, the diverse but effective asexual propagation capacity via cuttings at the species level, and summarize the results of a pilot cultivation trial set up in 2020 (still ongoing) outlining tree growth rates and the onset of fruit production among genotypes and species. Overall, the meta-analysis of previously published data in conjunction with new data generated herein may serve the sustainable exploitation of the studied NUPs.

Documenting Greek Indigenous Germplasm of Cornelian Cherry (Cornus mas L.) for Sustainable Utilization: Molecular Authentication, Asexual Propagation, and Phytochemical Evaluation.

Wild-growing Cornelian cherries (Cornus mas L., Cornaceae) are well-known native fruits in Greece since ancient times that are still consumed locally nowadays. Modern research has highlighted the value of Cornelian cherries as functional food with exceptional health benefits on account of the fruits' biochemical profile. However, apart from local consumption directly from wild growing individuals, Greek native C. mas populations have not yet been investigated or sustainably utilized. A multifaceted evaluation was conducted herein including authorized collection-documentation, taxonomic identification, and molecular authentication (DNA barcoding), asexual propagation via cuttings and phytochemical evaluation (multiple antioxidant profiling) of neglected and underutilized Greek native C. mas germplasm sources. Successive botanical expeditions resulted in the collection of 18 samples of genotypes from distant C. mas populations across different natural habitats in Greece, most of which were DNA fingerprinted for the first time. Asexual propagation trials revealed high variability in rooting frequencies among Greek genotypes with low (<25%), average (25−50%), and adequate propagation potential (>50%) using external indole-3-butyric acid (IBA) hormone application on soft- or hard-wood cuttings. The comparative phytochemical evaluation of the studied Greek genotypes showed significant potential in terms of antioxidant activity (>80% radical scavenging activity in 13 genotypes), but with variable phenolic content (47.58−355.46 mg GAE/100 g), flavonoid content (0.15−0.86 mg CE/100 g), and vitamin C content (1−59 mg AAE/100 g). The collected material is currently maintained under ex situ conservation for long-term monitoring coupled with ongoing pilot cultivation trials. The pivotal data create for the first time a framework for the sustainable utilization of Greek native C. mas germplasm as a superfood with significant agronomic potential.

Molecular Authentication, Phytochemical Evaluation and Asexual Propagation of Wild-Growing Rosa canina L. (Rosaceae) Genotypes of Northern Greece for Sustainable Exploitation.

Dogroses belong to a taxonomically difficult genus and family and represent important phytogenetic resources associated with high ornamental, pharmaceutical-cosmetic and nutritional values, thus suggesting a potentially high exploitation merit. Triggered by these prospects, wild-growing Rosa canina populations of Greece were selected for investigation and evaluation of their potential for integrated domestication. We collected ripe rosehips from Greek native wild-growing populations (samples from seven genotypes) for phytochemical analysis (total phenolics, total flavonoids, antioxidant activity and vitamin C content), leaf samples for DNA analysis using the ITS2 sequence (nine genotypes) and fresh soft-wood stem cuttings for propagation trials (seven genotypes). After evaluation of these materials, this study reports for the first-time distinct DNA-fingerprinted genotypes from Greece with interesting phytochemical profiles mainly in terms of Vitamic C content (up to 500.22 ± 0.15 mg of ascorbic acid equivalents/100 g of sample) as well as effective asexual propagation protocols for prioritized R. canina genotypes via cuttings. The latter highlights the importance of the levels of external hormone application (2000 ppm of indole-3-butyric acid), the effect of season (highly-effective spring trials) and genotype-specific differences in rooting capacities of the studied genotypes. All inclusive, this study offers new artificially selected material of Greek native R. canina with a consolidated identity and interesting phytochemical profile. These materials are currently under ex-situ conservation for further evaluation and characterization in pilot field studies, thus facilitating its sustainable exploitation for applications in the agro-alimentary, medicinal-cosmetic, and ornamental sectors.

Study of potent cytotoxic activity of Helleborus cyclophyllus Boiss against a human adenocarcinoma cell line.

Helleborus cyclophyllus Boiss is a rhizomatous plant species, with strong allelochemical properties, that has been used since ancient times for its therapeutic properties. In the present study we investigated the ability of an aqueous-soluble fraction of the methanol extract of H. cyclophyllus Boiss leaves, to induce apoptotic cell death on A549 human bronchial epithelial adenocarcinoma cells. A primary human lung fibroblasts' cell line was used as a model of normal-healthy cells for comparison. Cell morphology was examined after appropriate staining, cytotoxic activity of the extract was determined by the MTT assay, the type of cell death was analyzed by flow cytometry, confirmation of apoptosis was evaluated with the analysis of caspase-3, PARP1 by western blotting, while the chemical composition was assessed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). H. cyclophyllus Boiss extract was selectively active on A549 cells inducing significant morphological changes, even at low concentrations. Characteristic morphological alterations included the release of vesicular formations from A549 cell membranes (ectosomes), detachment of cells from their substrate, generation of a large vesicle into the cytoplasm (thanatosome) and the formation of apoptotic bodies. The selective apoptotic action on treated cells was also confirmed by biochemical criteria. Low concentrations, however, did not affect normal cells. The phytochemical analysis of the extract revealed the presence of cardiac glucosides, bufadienolides and phytoecdysteroids. To the best of our knowledge, the above-mentioned sequences of events leading selectively cancer cells to apoptosis, has not been reported before.

Determinants of depression among patients with epilepsy in Athens, Greece.

OBJECTIVE: Depression is common among patients with epilepsy. The aim of our study was twofold: to estimate the prevalence of a major depressive episode and to identify its determinants among patients with epilepsy treated in the largest Greek hospital in Athens. METHODS: All consecutive patients with epilepsy that visited the epilepsy outpatient clinic of Evangelismos General Hospital were invited to participate in the study. Ninety-four patients met our inclusion criteria. RESULTS: A diagnosis of a current major depressive episode was established in 21 out of 94 eligible to participate (22.3%) patients. Being a female was associated with a 19.68-fold increase in the odds of having a major depressive episode (95% CI 3.39-114.14, p=0.001); being unemployed was associated with a 6.46-fold increase in the odds of having a major depressive episode (95% CI 1.23-34.07, p=0.028), and each extra seizure experienced per month was associated with a 1.38-fold increase in the odds of having a major depressive episode (95% CI 1.03-1.85, p=0.031). CONCLUSION: Unemployment, female gender, and seizure control are important determinants of a major depression episode among patients with epilepsy.

Validation of the Greek version of the Neurological Disorders Depression Inventory for Epilepsy (NDDI-E).

The Neurological Disorders Depression Inventory for Epilepsy (NDDI-E) was developed as a screening tool for symptoms of major depressive episodes in people with epilepsy. Our study describes the development, validation, and psychometric properties of the Greek version of the NDDI-E. A consecutive sample of 101 patients with epilepsy, eligible to participate in the study, has been assessed using the Mini International Neuropsychiatric Interview version 5.0.0 and the NDDI-E. All patients had no major difficulties in understanding or answering the questions of the Greek version. Cronbach's alpha coefficient was 0.74. Receiver operating characteristic analysis showed an area under the curve of 91% (95% CI=83%-99%; SE: 0.040, p<0.001). At a cutoff score of greater than 15, the NDDI-E showed a sensitivity of 91%, a specificity of 81%, and a negative predictive value of 97%.