Ispitivanje antivirusne aktivnosti ekstrakata žalfije Salvia officinalis L. (Lamiaceae)

In the present study, we examined cytotoxicity and extracellular and intracellular antiviral activity of frac­tionated extracts of wild and cultivated sage Salvia officinalis L. (Lamiaceae) in vitro using the WISH-VSV model system. Extracts were obtained by fractionating depigmented ethanol extracts of sage plants with supercritical CO2 at different pressures. Cytotoxicity was determined by examining cellular morphology in situ with the aid of a colorimetric micromethod and by cell staining with trypan blue. The fraction of distilled cultivated sage obtained at CO2 pressure of 300 bars and temperature of 60°C (149/3) was the most cytotoxic, with CTD10 44 μg/ml. That of non-distilled cultivated sage obtained at CO2 pressure of 500 bars and temperature of 100°C (144/5) was the least toxic (CTD10 199 μg/ml). Moreover, 144/5 had an antiviral effect at the intracellular level: when added 5 hours before VSV infection, it caused 100% reduction of CPE at concentrations of 99.5 and 199.0 μg/ml; when added after virus penetration had occurred, the same concentrations caused 35 and 60% reduction, respectively. The obtained results indicate that antiviral activity of 144/5 involves inhibition of the early steps of the virus infective cycle without a direct virucidal effect. Abbreviations: WISH - human amnion epithelial cells, VSV - vesicular stomatitis virus, HSV - herpes simplex virus, CPE - cytopathic effect, IS - selectivity index, TCID50 - tissue culture infective dose, CTD10 - 10% cytotoxic concentrations.U radu je ispitivana antiviralna aktivnost različito frakcionisanih ekstrakata divlje i gajene žalfije Salvia officinalis L. (Lamiaceae) u in vitro uslovima koristeći WISH-VSV model sistem. Ekstrakti su dobijeni frakcionisanjem depigmentisanog etanolnog biljnog ekstrakta pod različitim pritiskom CO2. Citotoksičnost je određivana praćenjem ćelijske morfologije in situ, i bojenjem ćelija sa tripan plavim. Frakcija gajene žalfije dobijena na CO2 pritisku od 300 bara i temperaturi od 60°C (149/3) je pokazala najveću citotoksičnost (CTD10 44 μg/ml). Frakcija nedestilisane gajene žalfije dobijena na CO2 pritisku od 500 bara i temperaturi od 100°C (144/5) je pokazala najmanju toksičnost (CTD10 199 μg/ml). Takođe, frakcija 144/5 je pokazala i antiviralni efekat na intracelularnom nivou: kada se ćelije tretiraju 5 sati pre VSV infekcije, redukcija CPE je bila 100% na koncentraciji od 99.5 i 199.0 μg/ml; kada se ćelije tretiraju posle ulaska virusa u ćeliju na istim koncentarcijama redukcija CPE iz­nosi 35% odnosno 60%. Dobijeni rezultati ukazuju da frakcija 144/5 ima antiviralnu aktivnost koja se ostvaruje krozinhibiciju ranih stupnjeva viralne infekcije bezdirektnog virucidalnog efekta

Effects of selenium nanoparticles on pathogenic and probiotic food bacteria, and their influence on human intestinal cells

Selenium is an especially important element for the functioning of antioxidative enzymes, making it one of the essential micronutrients. Although selenium is required in extremely small amounts, often it is lacking from the organism. However, inorganic and organic sources of selenium are known to possess considerable toxicity. Recently, the nanoparticle form of selenium has been shown to exhibit lesser toxicity, and also better antimicrobial and other biological effects, than selenium-containing compounds. Nanotechnology has already found application in many fields, for improving material properties for various purposes, among them, in the process of improving food characteristics. It is possible that selenium nanoparticles can be applied as antimicrobial agents in food, and there has been growing interest in this area of research. We have synthesized selenium nanoparticles (SeNPs) by chemical reduction method, using sodium selenite, ascorbic acid, and bovine serum albumin. The antimicrobial activity of these particles has been assessed by minimum inhibitory concentration assay and colony-forming units count, on Listeria monocytogenes and Salmonella enterica, which are common bacterial food contaminants. As intestinal microbiota, especially probiotic bacteria, have been shown to play important role in organism functioning, we also tested how SeNPs influence Lactobacillus rhamosus and Lactobacillus plantarum. To evaluate cytotoxicity, we used MTT assay on the human intestinal cell line (HT-29). Antioxidative properties were assessed by DPPH assay. SeNPs showed higher antibacterial activity on pathogenic bacteria than on probiotic strains. Cytotoxic effect was present only in the highest tested concentrations. The antioxidative activity was considerable, resulting in above 80% reduction of DPPH free radical, at 1 g/ml of SeNPs. Based on these results, SeNPs are a promising subject for further research in developing materials for food packaging or as antimicrobial selenium additives

Cytotoxic and genotoxic potential of composite based on resveratrol and selenium nanoparticles, and its potential for superoxide anion induction

Kontinuirano interesovanje za nanočestice različitih materijala i njihove kompozite stvorilo je brojna pitanja u vezi njihove potencijalne toksičnosti. Polifenol resveratrol i jedinjenja sa selenom su među agensima čijoj je upotrebi nanotehnologija omogućila nov pristup. U formi nanočestica, oni pokazuju poboljšanu biokompatibilnost i biološke efekte, kao što je antimikrobna aktivnost. Na osnovu podataka iz literature, nanočestice selena su najčešće manje toksične nego jedinjenja sa selenom. U našem istraživanju, najpre smo pripremili kompozitni materijal na bazi selena i resveratrola. Nakon toga, kompozit i njegove komponente bili su testirani pomoću MTT tetrazolijumske boje za utvrđivanje citotoksičnosti, komet testom na genotoksičnost i nitroplavo-tetrazolijumskim testom za procenu nivoa superoksidnih anjona. U svim eksperimentima je korišćena zdrava ćelijska linija fibroblasta čoveka (MRC-5). Rezultati su pokazali da, iako čestice resveratrola ispoljavaju citotoksičnost u koncentracijama preko 12 μg/ml, one nisu genotoksične i ne indukuju stvaranje superoksidnih anjona. Sa druge strane, nanočestice selena nisu bile citotoksične ispod 150 μg/ml, ali su njima tretirane ćelije pokazale znake oštećenja DNK i povećanje produkcije superoksidnih anjona, čak i na 18 μg/ml. Kompozit je pokazao drugačije efekte, indukujući višu citotoksičnost i oksidativni stres nego ekvivalentne koncentracije komponenti, ali istovremeno nije bio genotoksičan. Moguće je da su interakcije nanočestica selena sa nanočesticama resveratrola dovele do pojave jedinstvenih osobina kompozita, potvrđujući izuzetnu važnost testiranja nanokompozitnih materijala uporedo sa njihovim komponentama.The continuous interest in engineered nanoparticles of various materials, and their composites, has raised numerous questions regarding their toxicological profiles. Polyphenol resveratrol and selenium compounds are among the agents to which nanotechnology has given all new approach. Their particulate forms possess improved bioavailability and biological effects, such as antibacterial activity. Furthermore, selenium nanoparticles usually have significantly lower toxicity compared to selenium compounds. In this study, we prepared composite material consisting of selenium and resveratrol nanoparticles. Then, composite material and its components were assessed by using tetrazolium MTT dye for determining cytotoxicity, comet assay for genotoxicity study, and nitroblue tetrazolium assay (NBT) assay to estimate levels of superoxide anions. The normal human fibroblast cells (MRC-5) were used in all the experiments. Results showed that, although the resveratrol component exhibited cytotoxicity at 12 μg/ml, it was not genotoxic, and did not induce superoxide anion production. Selenium nanoparticles, on the other hand, were not cytotoxic below 150 μg/ml, but treated cells showed signs of DNA damage and elevation in superoxide anion production even at 18 μg/ml. Composite material behaved differently, inducing higher cytotoxicity and oxidative stress than comparable concentrations of components, but also showed lack of genotoxicity. Possible interactions of selenium nanoparticles with resveratrol nanoparticles could have created unique composite properties, confirming the great importance of toxicity testing of nanocomposite materials along with their components

Synthesis and characterization of composite resveratrol/selenium nanomaterial, and preliminary assessment of its’ antioxidative effect and biocompatibility

Natural chemicals and earth elements are increasingly used in research as basis for novel materials intended for use in medicine. Among phytopharmaceuticals, more specifically polyphenols, resveratrol is known for its’ antioxidative, anticancer, antimicrobial, and other beneficial effects. Selenium, an essential trace element, is lately being recognized in nanoparticle form as less toxic and equally or more efficient than commercially available forms. The synergy of these two agents have not been shown until lately, when their synergistic antioxidative and gene expression effects were investigated for the purpose of treating Alcheimer disease. During our previous research, we have successfully synthesized pure resveratrol particles, as well as selenium nanoparticles (SeNPs). Both of them were separately investigated regarding their biological activities. The first step in evaluation of their possible synergistic antioxidative effect was obtaining the stable composite of these two materials. Synthesis parameters and processing methods were varied, and obtained suspensions assessed by their macroscopic and microscopic characteristics. The sample with both components homogeneously distributed in the particle form, was chosen for further experiments. Ultraviolet-visible (UV-Vis) spectrophotometry and Fourier transform infrared spectroscopy (FTIR) were used to characterize sample, and antioxidative activity (by DPPH reduction assay), and cytocompatibility (using MTT cytotoxicity assay) were additionally determined. Results showed improved cytocompatibility as compared to pure resveratrol particles, and preserved, significantly high antioxidative potential

Additive-free resveratrol micro- and nanoparticles and assessment of their toxicity

Resveratrol, polyphenol from stilbene family, has demonstrated beneficial effects on human health in numerous studies. Among these effects are cardioprotective, anti-cancer, neuroprotective, antimicrobial and other. In vivo, application of this phytochemical often lacks significant results. This is mainly because of its poor bioavailability, caused by low water solubility, sensitivity to light, oxygen and pH change, and high reactivity. Micro- and nanoformulations of resveratrol have been described to highly improve bioavailabiliy. However, there are limitations to this approach, such as demanding synthesis process, necessity of expensive or toxic chemicals, or poor loading capacity. We have used a simple physicochemical solvent-non solvent method to synthetise additive-free, stabile and uniform elongated resveratrol micro- and nanoparticles (RES-particles). The drying conditions were varied to examine their influence on the morphological characteristics of resveratrol particles. Differential effects of drying at ambient, low, and elevated temperature as well as effects of centrifugation and stability of RES-particle suspension during the time were documented. RES-particles were characterized by X-ray diffraction (XRD), differential scanning calorimetry (DSC), ultraviolet-visible (UV-Vis) spectrophotometry, Fourier transform infrared spectroscopy (FTIR) and optical microscopy. Biocompatibility of RESparticles was preliminary assessed through MTT toxicity assay

Antioxidative Resveratrol Particles as a Bioactive Component for Material Design

There is a growing need for multifunctional components in material design, especially in tissue engineering. Among many natural compounds, polyphenols are gaining interest as biologically active additives of novel materials. Stilbenoid polyphenol resveratrol is one of the most known, mainly because of its antioxidative, antibacterial, anticancer, neuroprotective and other beneficial effects. To allow most efficient use of resveratrol, several nanoformulations have shown promise, but most of them also involve other, toxic or expensive compounds, beside resveratrol. During our research, we synthetized particles of pure resveratrol, in an elongated, nanobelt-like form. The shape of the particles, as well as absence of any polymer or other carriers makes these particles easy to handle for further implementation. For preliminary assessment of the functionality of these particles, several assays were employed. DPPH, TBA and FRAP assay proved antioxidative potential of obtained particles by several mechanisms in the concentration range from 1 μg/mL to 10 μg/mL. Artemia salina in vivo bioassay was used to show high bioactivity and also biocompatibility of the particle suspension. All of the results indicated that there is a high potential for use of these res

Supplementary data for the article: Mitic-Culafic, D.; Nikolic, B.; Simin, N.; Jasnic, N.; Cetojevic-Simin, D.; Krstic, M.; Knezevic-Vukcevic, J. Effect of Allium Flavum L. and Allium Melanantherum Pan. Extracts on Oxidative DNA Damage and Antioxidative Enzymes Superoxide Dismutase and Catalase. Plant Food Hum. Nutr. 2016, 71 (1), 28–34. https://doi.org/10.1007/s11130-015-0519-0

Supplementary material for: [https://doi.org/10.1007/s11130-015-0519-0]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2065

Protective effect of Gentiana lutea root and leaf extracts against heterocyclic aromatic amines IQ and PhIP produced in thermally processed meat

During high-temperature cooking of protein rich foods, especially meat and fish, heterocyclic aromatic amines can be formed. These amines are a class of potent mutagens that can cause alterations in the structure of DNA and chromosomes. In recent decades, research has been focused on investigating plants and their phytochemicals as potential antimutagens. The aim of this study was to examine the anti-genotoxic effect of methanolic root and leaf extracts of Gentiana lutea against the food mutagens 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) produced in thermally processed meat. To determine the protective potential of extracts, the alkaline comet assay was applied. The results obtained indicated strong anti-genotoxic effect of both extracts against the tested mutagens. The highest inhibition of IQ-induced genotoxicity was recorded for leaf extract (72%). Regarding PhiP, root extract achieved inhibition of 80% of DNA damage, so was more successful than leaf extract. The data obtained in this study stimulates further research of G. lutea extracts and its constituents as potential dietary supplements in improving human health. © Published under licence by IOP Publishing Ltd.Conference of 60th International Meat Industry Conference, MEATCON 2019 ; Conference Date: 22-25 September 2019; Conference Code:15329

Cheese Fermented with Human-Derived Limosilactobacillus reuteri DSM 17938 and Mushroom Powders: A Novel Psychobiotic Food with Enhanced Bioactivity and Sensory Acceptability

Fermented foods containing psychobiotics are of growing interest among food scientists. Human-derived Limosilactobacillus reuteri DSM 17938, a gut symbiont and potential psychobiotic strain, has been shown to exhibit the following health benefits: anti-inflammation and GABA-production capacity, as well as modulation of pathogen and cancer cell growth. The aim of this research was to develop an acid-coagulated fresh soft quark-type cheese, fermented with L. reuteri DSM 17938, with enhanced bioactivity, sensory acceptability, and overall likeability. Psychobiotic-containing cheeses represent the food of a new generation, so it is of great importance to gain the trust of the consumers. To develop a familiar taste, cheese samples were enriched with mushroom powders of Agaricus bisporus and Pleurotus ostreatus. A high abundance of lactic acid bacteria was maintained in all cheese samples (>log 7.64 CFU/mL), while cheese extracts exhibited cytotoxicity to colon cancer cell line HCT116 (up to 30.96%) in vitro. Additionally, cheese samples provided a favorable medium for the growth of the probiotic Escherichia coli Nissle 1917 (>log 7.11 CFU/mL). Sensory evaluation revealed high scores for all samples (up to 97.21% of maximum overall quality). The survey conducted in this study offered insights into consumer willingness to try products containing psychobiotics. This study demonstrates the potential for the successful development of fermented food products with L. reuteri DSM 17938, which exhibits all the desired traits that consumers may receive well. Further research is required to explore the potential health benefits of these innovative food products