Pharmacognosy and Medicinal Plants

The anticancer activity of volatile isolates from Allium commutatum Guss., Allium sativum L. (Amaryllidaceae family), and Lepidium graminifolium L. ssp. graminifolium (Brassicaceae family), analyzed by GC-MS, as well as the pure sulfur volatiles, were evaluated against two human tumor cell lines: glioblastoma cell line LN-229 and bladder cancer cell line UM-UC-3, using MTT assay. The major sulfur volatiles found in A. commutatum hydrodistillate from flower and A. sativum bulb originated from S-alk(en)yl cysteine sulfoxide degradation. The most abundant sulfur volatile in A. commutatum isolate was dipropyl trisulfide, and in A. sativum diallyl disulfide. A. commutatum distillate showed very weak cytotoxic effect on both cancer cell lines except at incubation time of 72 h on LN229 cell line (IC50 6.364 µg/mL), while dipropyl trisulfide showed much stronger cytotoxic effect: with IC50 12.34 and 10.35 µg/mL, and IC50 22.19 and 8.434 µg/mL for UM-UC-3 and LN229 cell line, respectively during incubation time of 48 and 72 h. Both A. sativum extract and distillate showed strong time and concentration-dependent cytotoxic activity on both cancer cell lines, with the best results at incubation time of 48 and 72 h. A. sativum extract had IC50 14.49 and 12.48 µg/mL, and 40.84 and 10.41 µg/mL for UM-UC-3, and for LN229 cell lines, respectively, during incubation time of 48 and 72 h. A. sativum distillate showed very similar results: IC50 20.86 and 14.13 µg/mL, and 17.41 and 12.01 µg/mL for UM-UC-3, and for LN229 cell line, respectively. As expected, an active compound from A. sativum diallyl disulfide showed very strong anticancerogenic potential with IC50 22.3 and 19.07 µg/mL, and 44.69 and 8.85 µg/mL for UM-UC-3, and LN229 cell line, respectively. L. graminifolium extract and distillate sulfur volatiles originated from glucosinolates degradation i.e. 3-methoxybenzyl isothiocyanate, and benzyl isothiocyanate. They didn’t show strong cytotoxic activity on UM-UC-2 cell lines. While there was effect on LN229 cell line of distillate at incubation time of 48 hours (IC50 53.92 µg/mL) and of extract at incubation time of 48 and 72 hours (IC50 30.71 and 54.37 µg/mL, respectively). On the contrary, benzyl isothiocyanate showed much stronger cytotoxic effect: with IC50 13.16 and 12.3 µg/mL, and IC50 6.48 and 12.29 µg/mL for UM-UC-3, and LN229 cell line, respectively during incubation time of 48 and 72 h.

dimethylnitrosamine in rats. Carcinogenis 7:595–599. Biography Nikola Stojanovic obtained his MD Degree from the Department of Medicine (Faculty of Medicine, University of Niš, Serbia) and was awarded as the Best Graduated Student for the graduation year 2014/2015. He began his research work during the second year of his studies and he is now doing a large number of specialized in vivo and in vitro experiments in the fields of Pharmacology, Toxicology, Biology, Immunology and Microbiology. His main focus currently involves the effects of essential oils on the levels of anxiety in both humans and animals. Up to now, he had participated and won several prizes, on national and international congresses in different fields of research. Besides that, he is an Author and Coauthor of a number of publications in highly esteemed peer-reviewed journals.

Statement of the Problem: The essential oil of Melissa officinalis leaves is held in high esteem for its use in aromatherapy; however, due to the low yield of the essential oil, its production cost is very high. A myriad of beneficial biological activities of this essential oil was reported and its composition has been extensively studied. Strangely, up to date, no studies exist on the acute toxicity of this essential oil. Prompted by this, in this work, the toxicity of orally administered M. officinalis essential oil was assessed. Methodology & Theoretical Orientation: The hydrodistilled essential oil used in the current study was obtained from fresh plant material (leaves, yield 0.087%, w/w) and the detailed analyses (GC and GC/MS) showed that the tested essential oil contained high amounts of geranial (22.1%), neral (17.6%), citronellal (4.2%), nerol (1.3%) and geraniol (1.2%), as expected for M. officinalis (3). The acute toxicity was evaluated in female BALB/c mice that were orally treated with the essential oil (in the dose range 0.5-3 g/kg). During a 24-h period, the animals’ behavior was monitored, and after that, the survived animals were sacrificed and, in their sera, liver damage-related parameters were evaluated. Findings: Doses over 1 g/kg decreased animal movement, produced abdominal writhings, tumbling, atony, spastic movements and in some cases muscle rigidity. All these symptoms were dose dependent and could probably be brought in connection with the amount of citronellal in the applied doses. Serum levels of ALT and AST, as well as their ratio (AST/ALT), increased with the applied essential oil in doses>1 g/kg, indicating liver toxicity. Conclusion & Significance: In conclusion, by causing a wide panel of both behavioral alterations and changes in serum biochemical parameters in mice, M. officinalis essential oil can be deemed as being moderately toxic.

Lucía Bada is a Graduate in Pharmacy from the Universidade de Santiago de Compostela (Spain), 2015. She has a Master’s Degree in Research and Development of New Drugs. Her passion lies in natural products, especially medicinal plants. She is interested in medicinal and chemical properties of phyto-preparations and phytotherapy researches in order to assess their scientific value, promote their pharmacological valorization and stimulate sustainable drug development with a high socioeconomic potential that would enhance the welfare of communities.

In recent years, natural resources have regained great importance in the development and research of new pharmacological agents. “Sierra del Caurel” is a natural spot located in the heart of Galicia (Spain). It is presented as a potent reservoir of bioactive secondary metabolites. Dianthus hyssopifolius L. is an herbaceous plant belonging to the Caryophyllaceae family. It has been used as a medicinal plant for decades by the inhabitants of the “Sierra del Caurel” because of its antirrheumatic and anticatarrhal properties. However, to date, few phytochemical and pharmacological data have been reported on species of the genus Dianthus. Therefore, Dianthus hyssopifolius L. has been selected to study its composition and therapeutic activity. To obtain the secondary metabolites, extraction with organic solvents was first carried out. The whole plant was macerated fresh in methanol at room temperature. The crude extract was resolved in methanol:water (3:1) and extracted with hexane. The polar phase was evaporated under reduced pressure to methanol: water (1:5) after extraction with dicloromethane. After evaporation of the solvent, three oil extracts were obtained: hexane, dichloromethane and methanol. The methanol extract was solubilized in hot ethanol to yield a yellow precipitate. Sephadex LH-20 and silica gel column chromatography as well as HPLC were used in order to fractionate the precipitate and to purify the obtained compounds. Using one- and two-dimensional NMR experiments and mass spectrometry, two flavonoids have been identified: a glycoside derived from quercetin and another from kaempferol. According to the bibliography review, study of antibacterial, anticancer, antifungal, anti-inflammatory, and antioxidant properties for the different extracts are currently in progress.

Sesquiterpenes are C15-terpenoids built from three isoprene units. These compounds are commonly found in higher plants, marine organisms and fungi. Naturally, they occur as hydrocarbons or in oxygenated forms including lactones, alcohols, acids, aldehydes, and ketones. Over the last decade, sesquiterpenes have attracted significant attention because of the roles they play in biological systems and their diverse biological activities, such as anti- inflammatory, antibacterial, antioxidant and anti-carcinogenic. The antiproliferative activity of three sesquiterpenes (thuyopsene, nerolidol and farnesol) was evaluated on three lines of human bladder cancer (T24, UM-UC-3, TCCSUP,) using MTT assay. Thuyopsene is one of the major sesquiterpene constituents of cedarwood, while farnesol and nerolidol can be found in the essential oils of many types of plants and flowers. The tested compounds showed different antiproliferative activity on the cancer cell lines. Farnesol showed the highest impact on T24 cells growth (52% growth inhibition) at concentration of ϱϲ μM, while thuyopsene showed the strongest growth inhiďition of ϱϬ% and 27% on UM-UC-3 and TCCSUP cell line at concentration of ϭϳϭ and ϭϮϮ μM, respectively. Generally, nerolidol showed the lowest antiproliferative potential on the cancer cells. In order to determine type of cell death induced by treatment, Annexin-V-FITC assay was used for the detection of apoptosis by flow cytometry. The results obtained for these three sesquiterpenes open a perspective for their use as antiproliferative agents.

Alexandra Bodó is a PhD student at the University of Pécs, Hungary. She has been researching on honey for a couple of years, first as a beekeeper, later as a researcher. She finds it important to investigate the quality and bioactivity of honeys, in order to avoid adulterations and provide customers with honeys of certified origin and beneficial health effects. Besides the research work, she takes part in teaching plant anatomy and plant physiology courses.

Honey is a supersaturated solution of sugars, which contains phenolic compounds, enzymes, free amino acids, minerals, vitamins and proteins acting as minor components. It is well established that honey has antioxidant effect, for which the phenolic acids and flavonoids are responsible. The aim of this study was to evaluate the antioxidant capacity of some floral and honeydew honeys available on the Hungarian market, with four different antioxidant techniques; and to classify the honeys according to melissopalynological analysis. We applied spectrophotometric methods: Folin-Ciocalteu (FC) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay for antiradical activity, trolox equivalent antioxidant capacity (TEAC) assay for total antioxidant activity, ABS450 for color intensity and one fluorimetric method: oxygen reactive antioxidant capacity (ORAC) for the antilipoperoxidant activity. To classify the botanical origin of honeys, we carried out qualitative and quantitative melissopalynological analyses. The antioxidant capacity measured by the Folin reagent reactivity correlated with the color parameters, as well as with the antioxidant activities measured with the other methods. Black locust honey samples with pale color had the lowest (116 mAU), while dark honeydew honeys called “meadow clary (Salvia pratensis) honey” exhibited the highest (1617 mAU) radical scavenging activity. Values ranged from 197.93±4.64 to 937.64±28.43 mg GAE/kg, 89.14±26.9 to 240.15±47.3 µM TE/100g, 61.76±2.85 to 5.47±0.02, and 14.78±1.15 to 114.89±10.43 µM TE/g for the FC, TEAC, DPPH (IC50), and ORAC assays, respectively. However, the melissopalynological analysis revealed that the “meadow clary honey” was in fact of multifloral origin. Among unifloral honeys the chestnut (Castanea sativa) honeys (1037 mAU) had the highest antioxidant activity (PC: 636.48±31.9 mg GAE/kg, TEAC: 171.23±2.0 µM TE/100g, DPPH (IC50): 17.37±0.57, ORAC: 75.2±4.71 µM TE/g). The results of this study demonstrated that the botanical origin and the color intensity of honeys have the greatest influence on their antioxidant activity.

Maria J Rodrigues has a Degree in Biology (Major in Biomedicine) and a Master’s in Biomedical Sciences. She has interest in exploring new biotechnological applications and products from marine organisms (halophytes, microalgae, macroalgae and marine invertebrates) found in the Algarve coast. She has been working as a Research Fellow in the Marine Biotechnology group under the scope of different projects: SEABIOMED (marine photosynthetic organisms of the Algarve coast with biomedical applications, XtremeBio (halophytes: a precious resource nutritional elements and bioactive compounds, MaNaCruzi (searching for marine origin molecules against Trypanosoma cruzi, and recently Xtreme Gourmet (extremophile plants in the gourmet cuisine). Since 2014, she has been working on her PhD project: "Unravelling the biotechnological potential of halophytes species of the Algarve coast". During this time, she participated in the preparation of 27 scientific articles published in international peer-reviewed journals and 17 communications in national and/or international conferences (oral and poster).

The growing interest of stakeholders and consumers in herbal products urges the need for research aiming to unravel innovative products with health improvement potential. Green tea (Camellia sinensis (L.) Kuntze, CS) is commonly sold in mixtures combining different herbs, fruits or spices in order to improve or add beneficial properties to such beverages. Having this in mind, and following previous promising results obtained with the halophyte sea lavender (Limonium algarvense Erben. LA) this work prepared and evaluated herbal beverages containing mixtures of CS and LA flowers in terms of antioxidant capacity and inhibition of enzymes related with Alzheimer’s (acetyl- and butyrylcholinesterase) and Type 2 diabetes mellitus (α-amylase and α-glucosidase). The phenolic profile was determined by HPLC. Both synergistic and antagonistic interactions were observed. LA and CS samples had strong antioxidant activity, whereas LA and CS mixtures exhibited higher OH radical-scavenging and anti-lipid peroxidation capacity. LA samples had higher cholinesterase inhibition than CS and mixtures resulted in stronger enzymatic inhibition. CS had the highest α-glucosidase inhibition, which decreased when combined with LA. CS had higher phenolic contents, and its combination with LA increased the phenolic diversity of the mixtures. Results showed that LA and CS infusions and decoctions and their combinations have relevant in vitro antioxidant, neuroprotective and antidiabetic properties. Infusions and decoctions of LA and CS mixtures should thus be further explored as potential innovative functional beverages able to prevent oxidative stress and lipid oxidation related diseases, and to reduce the progression of neurodegenerative diseases and diabetic complications.

En Hyung Kim finished her residency in Dermatology at Ajou University Hospital and is currently the Director of Department of Dermatology at Cheil General Hospital and Women’s Healthcare Center. As a Scientist, she is interested in Skin Physiology and Inflammatory Mechanisms. As a Clinician, she makes effort to find new innovative ways to treat her patients effectively and safely.

Ginsenosides are representative pharmaceutical compounds found in various forms in Panax ginseng, a traditional medicinal plant. There have been many reports describing the biological activities, including anti-inflammatory, anti-tumor, and anti-dementia effects, of several ginsenosides. The biological actions of these ginsenosides have been closely related to their biotransformations by intestinal microbiome. They are converted to their metabolites Rg2, Rg3, compound K, and others by human intestinal microflora following ingestion. The main functional component detected in mammalian blood or organs after oral administration of ginseng or ginsenosides is compound K. Compound K has been reported to exhibit diverse biological functions, including antitumor, antidiabetic, antiallergic, and anti-inflammatory effects in vitro and in vivo. Recently, antiaging effects of ginsenosides in human skin have been reported from clinical trial and in vitro model data. Ginsenosides have hence been proposed as promising natural cosmeceutical agents. We reviewed the biotransformation and delivery of compound K. Also biological effects of ginsenosides, especially compound K, on skin health and its potential use as cosmeceutical agents was studied