Assessment of in vitro antioxidant properties of medicinal and edible plant extracts

Identification and characterization of plant-based products with antioxidant and anti-proliferative effects has received much interest over the past few years as possible therapeutic mean for treatment of diseases likely to be associated to oxidative stress (such as atherosclerosis, diabetes, and cancer). This study provides characterization of in vitro antioxidant and/or anti-proliferative potential and phytochemical profile of (i) 39 wine samples of underutilized Georgian grapevine cultivars, (ii) extracts of 22 samples of medicinal plants from Ethiopia and (iii) 23 samples of edible and medicinal plants from Peruvian Amazon. For this purpose, 2,2-diphenyl-1-picrylhydrazyl radical assay (DPPH), oxygen radical absorbance capacity (ORAC) assay, total phenolic content (TPC), and cell viability assay based on metabolization of tetrazolium bromide (MTT) to formazan, together with methods based on high performance liquid chromatography (HPLC-ultra violet/visible spectrometry and HPLC-tandem mass spectrometry) were used. Georgian red wines (particularly Saperavi cultivars) exhibited higher antioxidant capacity (average DPPH and ORAC values at 5.1 and = 10.6 g TE/L wine, respectively) in comparison to Central and West European cultivars [Pinot Noir (DPPH = 3.1 and ORAC = 9.4 g TE/L wine), Cabernet Sauvignon (DPPH = 3.0 and ORAC = 7.3 g TE/L wine) and Cabernet Moravia (DPPH = 2.0 and ORAC = 8.5 g TE/L wine)]. Georgian wines contained significantly greater concentrations of quercetin (between 14.44 and 1.07 ug/mL), kaempferol (between 1.68 and 0.03 ug/mL) and syringic acid (between 12.59 and 4.72 ug/mL), whereas possessed lower quantities of resveratrol (between 5.11 and 0.32 ug/mL) in comparison to Central and West European wines. Amongst edible and medicinal plants from Ethiopia and Peruvian Amazon, only Dodonaea angustifolia (IC50 for DPPH = 22.2 ug/mL, ORAC = 767.6 ug TE/mg extract; IC50 for Hep-G2 = 120.0 ug/mL), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract; IC50 for Hep-G2 = 50.5 ug/mL), Inga edulis (DPPH and ORAC = 337.0 and 795.7 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 36.3 and 57.9 ug/mL) and Oenocarpus bataua (DPPH and ORAC = 903.8 and 1024.4 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 102.6 and 38.8 ug/mL) have demonstrated combinatory antioxidant/anti-proliferative efficacy. Selective anti-proliferative activity was observed for Verbascum sinaiticum (IC50 for Hep-G2 = 80.6 ug/mL) and Annona montana (IC50 for Hep-G2 and HT-29 = 2.7 and 9.0 ug/mL, respectively). Above-mentioned plant material showed only weak or non-toxic effects towards normal cell line. Despite the fact that extracts of Jasminum abyssinicum (IC50 for DPPH = 26.3 ug/mL, ORAC = 1023.7 ug TE/mg extract), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract), Mauritia flexuosa (DPPH and ORAC = 1062.9 and 645.9 ug TE/mg extract), Myrciaria dubia (DPPH and ORAC = 641.9 and 642.6 ug TE/mg extract) and Theobroma grandiflorum (DPPH and ORAC = 714.8 and 821.9 ug TE/mg extract) have exhibited considerable antioxidant effect, these species were found to possess moderate to low anti-proliferative potential or have shown to be toxic to normal cells line. In all cases it was detected that phenolic compounds content correlated strongly with antioxidant activity, however weakly with anti-proliferative effect. Results suggest above-mentioned species as prospective materials for further development of novel plant-based agents effective against oxidative stress related diseases. However, it is necessary to perform further research which would be focused on detailed characterization of their chemical composition, pharmacological effects and toxicological safety, in order to verify their possible practical use

Assessment of in vitro antioxidant and anti-proliferative properties of medicinal and edible plants

Identification and characterization of plant-based products with antioxidant and anti-proliferative effects has received much interest over the past few years as possible therapeutic mean for treatment of diseases likely to be associated to oxidative stress (such as atherosclerosis, diabetes, and cancer). This study provides characterization of in vitro antioxidant and/or anti-proliferative potential and phytochemical profile of (i) 39 wine samples of underutilized Georgian grapevine cultivars, (ii) extracts of 22 samples of medicinal plants from Ethiopia and (iii) 23 samples of edible and medicinal plants from Peruvian Amazon. For this purpose, 2,2-diphenyl-1-picrylhydrazyl radical assay (DPPH), oxygen radical absorbance capacity (ORAC) assay, total phenolic content (TPC), and cell viability assay based on metabolization of tetrazolium bromide (MTT) to formazan, together with methods based on high performance liquid chromatography (HPLC-ultra violet/visible spectrometry and HPLC-tandem mass spectrometry) were used. Georgian red wines (particularly Saperavi cultivars) exhibited higher antioxidant capacity (average DPPH and ORAC values at 5.1 and = 10.6 g TE/L wine, respectively) in comparison to Central and West European cultivars [Pinot Noir (DPPH = 3.1 and ORAC = 9.4 g TE/L wine), Cabernet Sauvignon (DPPH = 3.0 and ORAC = 7.3 g TE/L wine) and Cabernet Moravia (DPPH = 2.0 and ORAC = 8.5 g TE/L wine)]. Georgian wines contained significantly greater concentrations of quercetin (between 14.44 and 1.07 ug/mL), kaempferol (between 1.68 and 0.03 ug/mL) and syringic acid (between 12.59 and 4.72 ug/mL), whereas possessed lower quantities of resveratrol (between 5.11 and 0.32 ug/mL) in comparison to Central and West European wines. Amongst edible and medicinal plants from Ethiopia and Peruvian Amazon, only Dodonaea angustifolia (IC50 for DPPH = 22.2 ug/mL, ORAC = 767.6 ug TE/mg extract; IC50 for Hep-G2 = 120.0 ug/mL), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract; IC50 for Hep-G2 = 50.5 ug/mL), Inga edulis (DPPH and ORAC = 337.0 and 795.7 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 36.3 and 57.9 ug/mL) and Oenocarpus bataua (DPPH and ORAC = 903.8 and 1024.4 ug TE/mg extract; IC50 for Hep-G2 and HT-29 = 102.6 and 38.8 ug/mL) have demonstrated combinatory antioxidant/anti-proliferative efficacy. Selective anti-proliferative activity was observed for Verbascum sinaiticum (IC50 for Hep-G2 = 80.6 ug/mL) and Annona montana (IC50 for Hep-G2 and HT-29 = 2.7 and 9.0 ug/mL, respectively). Above-mentioned plant material showed only weak or non-toxic effects towards normal cell line. Despite the fact that extracts of Jasminum abyssinicum (IC50 for DPPH = 26.3 ug/mL, ORAC = 1023.7 ug TE/mg extract), Rumex nepalensis (IC50 for DPPH = 5.7 ug/mL, ORAC = 1061.4 ug TE/mg extract), Mauritia flexuosa (DPPH and ORAC = 1062.9 and 645.9 ug TE/mg extract), Myrciaria dubia (DPPH and ORAC = 641.9 and 642.6 ug TE/mg extract) and Theobroma grandiflorum (DPPH and ORAC = 714.8 and 821.9 ug TE/mg extract) have exhibited considerable antioxidant effect, these species were found to possess moderate to low anti-proliferative potential or have shown to be toxic to normal cells line. In all cases it was detected that phenolic compounds content correlated strongly with antioxidant activity, however weakly with anti-proliferative effect. Results suggest above-mentioned species as prospective materials for further development of novel plant-based agents effective against oxidative stress related diseases. However, it is necessary to perform further research which would be focused on detailed characterization of their chemical composition, pharmacological effects and toxicological safety, in order to verify their possible practical use

Medicinal Use of Testosterone and Related Steroids Revisited

Testosterone derivatives and related compounds (such as anabolic-androgenic steroids—AAS) are frequently misused by athletes (both professional and amateur) wishing to promote muscle development and strength or to cover AAS misuse. Even though these agents are vastly regarded as abusive material, they have important pharmacological activities that cannot be easily replaced by other drugs and have therapeutic potential in a range of conditions (e.g., wasting syndromes, severe burns, muscle and bone injuries, anemia, hereditary angioedema). Testosterone and related steroids have been in some countries treated as controlled substances, which may affect the availability of these agents for patients who need them for therapeutic reasons in a given country. Although these agents are currently regarded as rather older generation drugs and their use may lead to serious side-effects, they still have medicinal value as androgenic, anabolic, and even anti-androgenic agents. This review summarizes and revisits the medicinal use of compounds based on the structure and biological activity of testosterone, with examples of specific compounds. Additionally, some of the newer androgenic-anabolic compounds are discussed such as selective androgen receptor modulators, the efficacy/adverse-effect profiles of which have not been sufficiently established and which may pose a greater risk than conventional androgenic-anabolic agents

Advances in the Determination of Anabolic-Androgenic Steroids: From Standard Practices to Tailor-Designed Multidisciplinary Approaches

Anabolic-androgenic steroids (AASs), a group of compounds frequently misused by athletes and, unfortunately, also by the general population, have lately attracted global attention; thus, significant demands for more precise, facile, and rapid AAS detection have arisen. The standard methods ordinarily used for AAS determination include liquid and gas chromatography coupled with mass spectrometry. However, good knowledge of steroid metabolism, pretreatment of samples (such as derivatization), and well-trained operators of the instruments are required, making this procedure expensive, complicated, and not routinely applicable. In the drive to meet current AAS detection demands, the scientific focus has shifted to developing novel, tailor-made approaches leading to time- and cost-effective, routine, and field-portable methods for AAS determination in various matrices, such as biological fluids, food supplements, meat, water, or other environmental components. Therefore, herein, we present a comprehensive review article covering recent advances in AAS determination, with a strong emphasis on the increasingly important role of chemically designed artificial sensors, biosensors, and antibody- and fluorescence-based methods

Vapors of Volatile Plant-Derived Products Significantly Affect the Results of Antimicrobial, Antioxidative and Cytotoxicity Microplate-Based Assays

Volatile plant-derived products were observed to exhibit broad spectrum of biological effects. However, due to their volatility, results of conventional microplate-based bioassays can be significantly affected by the vapors. With aim to demonstrate this phenomenon, antimicrobial, antioxidant, and cytotoxic activities of three essential oils (Alpinia elegans, Cinnamomum iners, and Xanthostemon verdugonianus), one supercritical CO2 extract (Nigella sativa), and four plant-derived compounds (capsaicin, caryophyllene oxide, 8-hydroxyquinoline, and thymoquinone) were evaluated in series of experiments including both ethylene vinyl acetate (EVA) Capmat sealed and nonsealed microplates. The results clearly illustrate that vapor transition to adjoining wells causes false-positive results of bioassays performed in nonsealed microtiter plates. The microplate layout and a duration of the assay were demonstrated as the key aspects defining level of the results affection by the vapors of volatile agents. Additionally, we reported biological activities and chemical composition of essential oils from A. elegans seeds and X. verdugonianus leaves, which were, according to our best knowledge, analyzed for the first time. Considering our findings, certain modifications of conventional microplate-based assays are necessary (e.g., using EVA Capmat as vapor barrier) to obtain reliable results when biological properties of volatile agents are evaluated

In vitro antioxidant and anti-proliferative activity of Ethiopian medicinal plant extracts

Identification and characterization of natural products with antioxidant and anti-proliferative activity has received much interest over the past few years. Ethiopia is one of the developing countries which have enormous diversity of plants and yet majority stays scientifically neglected and undiscovered. In this study, the ethanol extracts of 18 Ethiopian wild medicinal plants were investigated for their in vitro antioxidant and anti-proliferative potential. For this purpose DPPH (2,2-diphenyl-1-picrylhydrazyl), ORAC (oxygen radical absorbance capacity) and TPC (total phenolic content) assays together with MTT cell viability assay (performed on Hep-G2 and MRC-5) were used. Extracts of Carissa spinarum, Dodonaea angustifolia, Jasminum abyssinicum, Rumex nepalensis, Rubus steudneri and Verbascum sinaiticum exhibited the most significant results. However, it was discovered that C. spinarum, J. abyssinicum and R. steudneri possessed considerable toxicity against normal MRC-5 cell line. Only extracts of D. angustifolia and R. nepalensis demonstrated significant combinatory antioxidant/anti-proliferative effect, while V. sinaiticum showed best selective anti-proliferative activity. Since aforementioned extracts also exerted low or minimal toxicity to normal cells, we suggest these as prospective material for further development of novel plant-based agents effective against oxidative stress related diseases

In vitro immunomodulatory activity, cytotoxicity and chemistry of some central European polypores

Context: Some mushrooms of the order Polyporales are known for their immunomodulatory actions. Objective: The objective of this study is to evaluate the in vitro phagocytic and cytotoxic effects of extracts from polyporales native to Central Europe. Materials and methods: The effects of ethanol extracts from 27 polypore species on opsonized zymosan-induced phagocytosis of isolated human neutrophils were tested by a chemiluminescence method. Colon epithelial cell lines, Caco-2 and HT-29, were used for cytotoxicity assays, and extracts were chemically characterized in terms of total phenolic and β-glucan content. Results: We observed phagocytosis or respiratory burst enhancing activity in 17 extracts, of which five species, namely Aurantiporus fissilis (Berk. & M.A. Curtis) H. Jahn ex Ryvarden, Trametes gibbosa (Pers.) Fr., Piptoporus betulinus (Bull.) P. Karst, Neolentinus lepideus (Fr.) Redhead & Ginns, Polyporus squamosus (Huds.) Fr., significantly increased phagocytosis in granulocytes by 205, 181, 158, 155 and 141%, respectively. The β-glucan content of the three most potent extracts was 58, 42 and 74 mg/g, respectively, and the polyphenol content was 155.6, 133.5 and 155.2 μmol of gallic acid equivalent/g, respectively. Some extracts showed cytotoxic activity, with higher cytotoxicity in Caco-2 than in HT-29 cells. Pycnoporus cinnabarinus (Jacq.) P. Karst. extract was cytotoxic to both cell lines, with IC50 values of 81 and 31 μg/mL, respectively. Discussion and conclusion: The most promising extracts were from N. lepideus and Polyporus squamosus, which are edible species and may be considered safe. Our findings support their use as culinary preparations or food supplements for various immunological gut disorders

Synthetic polyploidization induces enhanced phytochemical profile and biological activities in Thymus vulgaris L. essential oil

Abstract Essential oil from Thymus vulgaris L. has valuable therapeutic potential that is highly desired in pharmaceutical, food, and cosmetic industries. Considering these advantages and the rising market demand, induced polyploids were obtained using oryzalin to enhance essential oil yield. However, their therapeutic values were unexplored. So, this study aims to assess the phytochemical content, and antimicrobial, antioxidant, and anti-inflammatory activities of tetraploid and diploid thyme essential oils. Induced tetraploids had 41.11% higher essential oil yield with enhanced thymol and γ-terpinene content than diploid. Tetraploids exhibited higher antibacterial activity against all tested microorganisms. Similarly, in DPPH radical scavenging assay tetraploid essential oil was more potent with half-maximal inhibitory doses (IC50) of 180.03 µg/mL (40.05 µg TE/mg) than diploid with IC50 > 512 µg/mL (12.68 µg TE/mg). Tetraploids exhibited more effective inhibition of in vitro catalytic activity of pro-inflammatory enzyme cyclooxygenase-2 (COX-2) than diploids at 50 µg/mL concentration. Furthermore, molecular docking revealed higher binding affinity of thymol and γ-terpinene towards tested protein receptors, which explained enhanced bioactivity of tetraploid essential oil. In conclusion, these results suggest that synthetic polyploidization using oryzalin could effectively enhance the quality and quantity of secondary metabolites and can develop more efficient essential oil-based commercial products using this induced genotype

Antioxidant and Anti-Inflammatory Activity of Five Medicinal Mushrooms of the Genus <i>Pleurotus</i>

Within the group of higher fungi, edible medicinal mushrooms have a long history of being used as food and in folk medicine. These species contain biologically active substances with many potential beneficial effects on human health. The Pleurotus genus is representative of medicinal mushrooms because Pleurotus ostreatus is one of the most commonly cultivated culinary mushrooms. In our study, we focused on lesser-known species in the genus Pleurotus and measured their antioxidant and anti-inflammatory activity. We prepared extracts of the mushrooms and analyzed them using HPLC−HRMS, GC−MS, and 1H-NMR. Significant differences in biological activities were found among the Pleurotus spp. extracts. A MeOH extract of P. flabellatus was the most active as a radical scavenger with the highest ORAC, while a chloroform extract had significant anti-inflammatory COX-2 activity. The 80% MeOH extract of P. flabellatus contained the highest amounts of ergosterol, ergothioneine, and mannitol. The 80% MeOH extract of P. ostreatus Florida was the most active in the NF-κB inhibition assay and had the highest content of β-glucans (43.3% by dry weight). Given the antioxidant and anti-inflammatory properties of P. flabellatus, the potential therapeutic usefulness of this species is worth evaluating through in-depth investigations and confirmation by clinical trials