Ivana Arsić

Anti-inflammatory, gastroprotective, free-radical-scavenging, and antimicrobial activities of hawthorn berries ethanol extract.

Hawthorn [Crataegus monogyna Jacq. and Crataegus oxyacantha L.; sin. Crataegus laevigata (Poiret) DC., Rosaceae] leaves, flowers, and berries are used in traditional medicine in the treatment of chronic heart failure, high blood pressure, arrhythmia, and various digestive ailments, as well as geriatric and antiarteriosclerosis remedies. According to European Pharmacopoeia 6.0, hawthorn berries consist of the dried false fruits of these two species or their mixture. The present study was carried out to test free-radical-scavenging, anti-inflammatory, gastroprotective, and antimicrobial activities of hawthorn berries ethanol extract. Phenolic compounds represented 3.54%, expressed as gallic acid equivalents. Determination of total flavonoid aglycones content yielded 0.18%. The percentage of hyperoside, as the main flavonol component, was 0.14%. With respect to procyanidins content, the obtained value was 0.44%. DPPH radical-scavenging capacity of the extract was concentration-dependent, with EC50 value of 52.04 microg/mL (calculation based on the total phenolic compounds content in the extract). Oral administration of investigated extract caused dose-dependent anti-inflammatory effect in a model of carrageenan-induced rat paw edema. The obtained anti-inflammatory effect was 20.8, 23.0, and 36.3% for the extract doses of 50, 100, and 200 mg/kg, respectively. In comparison to indomethacin, given in a dose producing 50% reduction of rat paw edema, the extract given in the highest tested dose (200 mg/kg) showed 72.4% of its activity. Gastroprotective activity of the extract was investigated using an ethanol-induced acute stress ulcer in rats with ranitidine as a reference drug. Hawthorn extract produced dose-dependent gastroprotective activity (3.8 +/- 2.1, 1.9 +/- 1.7, and 0.7 +/- 0.5 for doses of 50, 100, and 200 mg/kg, respectively), with the efficacy comparable to that of the reference drug. Antimicrobial testing of the extract revealed its moderate bactericidal activity, especially against gram-positive bacteria Micrococcus flavus, Bacillus subtilis, and Lysteria monocytogenes, with no effect on Candida albicans. All active components identified in the extract might be responsible for activities observed.

Anti-inflammatory, gastroprotective, and cytotoxic effects of Sideritis scardica extracts.

Sideritis scardica Griseb. (ironwort, mountain tea), an endemic plant of the Balkan Peninsula, has been used in traditional medicine in the treatment of gastrointestinal complaints, inflammation, and rheumatic disorders. This study aimed to evaluate its gastroprotective and anti-inflammatory activities. Besides, continuously increasing interest in assessing the role of the plant active constituents preventing the risk of cancer was a reason to make a detailed examination of the investigated ethanol, diethyl ether, ethyl acetate, and N-butanol extracts regarding cytotoxicity. Oral administration of the investigated extracts caused a dose-dependent anti-inflammatory effect in a model of carrageenan-induced rat paw edema. Gastroprotective activity of the extracts was investigated using an ethanol-induced acute stress ulcer in rats. The cytotoxic activity of plant extracts was assessed on PBMC, B16, and HL-60 cells and compared to the cytotoxicity of phenolic compounds identified in extracts. Apoptotic and necrotic cell death were analyzed by double staining with fluoresceinisothiocyanate (FITC)-conjugated annexin V and PI. The developed HPLC method enabled qualitative fingerprint analysis of phenolic compounds in the investigated extracts. Compared to the effect of the positive control, the anti-inflammatory drug indomethacine (4 mg/kg), which produced a 50 % decrease in inflammation, diethyl ether and N-butanol extracts exhibited about the same effect in doses of 200 and 100 mg/kg (53.6 and 48.7 %; 48.4 and 49.9 %, respectively). All investigated extracts produced dose-dependent gastroprotective activity with the efficacy comparable to that of the reference drug ranitidine. The diethyl ether extract showed significant dose-dependent cytotoxicity on B16 cells and HL-60 cells, decreasing cell growth to 51.3 % and 77.5 % of control, respectively, when used at 100 µg/mL. It seems that phenolic compounds (apigenin, luteolin, and their corresponding glycosides) are responsible for the diethyl ether extract cytotoxic effect. It also appears that induction of oxidative stress might be involved in its cytotoxicity, since B16 and HL-60 cells increased their ROS production in response to treatment with diethyl ether extract. Neither of the tested extracts nor any phenolic compounds showed significant cytotoxic effect to human PBMC. These results demonstrated the potent anti-inflammatory and gastroprotective activities, as well as the promising cytotoxicity.

Preparation of novel apigenin-enriched, liposomal and non-liposomal, antiinflammatory topical formulations as substitutes for corticosteroid therapy.

Two oil‐in‐water formulations, containing equal amounts of apigenin‐enriched chamomile flower extracts, for potential use as topical antiinflammatory agents, were prepared and their physicochemical properties evaluated. A pilot clinical study was then carried out to assess patient acceptability and efficacy. The creams were either non‐liposomal or liposomal. The liposomal formulations were more viscous, thus producing superior release characteristics in vitro. The clinical study also showed that the liposomal creams were, as antiinflammatory agents, slightly more effective in vivo than the non‐liposomal formulations. These results suggest that there is scope for the further development of even more effective and safer alternatives to corticosteroids. Copyright

Chemical and Antimicrobial Evaluation of Supercritical and Conventional Sideritis scardica Griseb., Lamiaceae Extracts

Sideritis scardica Griseb., Lamiaceae (ironwort, mountain tea), an endemic plant of the Balkan Peninsula, has been used in traditional medicine in the treatment of antimicrobial infections, gastrointestinal complaints, inflammation and rheumatic disorders. This study reports a comparison between conventional (hydrodistillation HD and solvent extraction SE) and alternative (supercritical carbon dioxide SC CO2) extraction methods regarding the qualitative and quantitative composition of the obtained extracts as analyzed by GC and GC-MS techniques and their anitimicrobial activity. Different types of extracts were tested, the essential oil EO obtained by HD, EO-CO2 and AO-CO2 obtained by SC CO2 at different preasures 10 and 30 MPa, at 40 °C, respectively, and the fractions A, B, C and D obtained by successive solvent extraction (SE) A: ethanol, B: diethyl ether, C: ethyl acetate and D: n-butanol). While EO was characterized by the presence of the high percentage of oxygenated monoterpenes and sesquiterpenes (30.01 and 25.54%, respectively), the rest of the investigated samples were the most abundant in fatty acids and their esters and diterpenes (from 16.72 to 71.07% for fatty acids and their esters, and from 23.30 to 72.76%, for diterpenes). Microbial susceptibility tests revealed the strong to moderate activity of all investigated extracts against the tested microorganisms (MIC from 40 to 2,560 μg/mL). Although differences in the chemical compositions determined by GC and GC-MS analysis were established, the displayed antimicrobial activity was similar for the all investigated extracts.

Lactobionic acid as antioxidant and moisturizing active in alkyl polyglucoside‐based topical emulsions: the colloidal structure, stability and efficacy evaluation

Cosmeceutical antioxidants may protect the skin against oxidative injury, involved in the pathogenesis of many skin disorders. However, an unsuitable topical delivery system with compromising safety profile can affect the efficacy of an antioxidant active. This study investigated the antioxidant potential of lactobionic acid (LA), a newer cosmeceutical active, per se (in solution) and incorporated into natural alkyl polyglucoside (APG) emulsifier‐based system using 1,1‐diphenyl‐2‐picrylhydrazyl free radical scavenging and lipid peroxidation inhibition assays. The α‐tocopherol was used as a reference compound. The physical stability (using rheology, polarization microscopy, pH and conductivity measurements) of an Alkyl glucoside‐based emulsion was evaluated with and without the active (LA); colloidal structure was assessed using polarization and transmission electron microscopy, rheology, thermal and texture analysis. Additionally, the safety profile and moisturizing potential were investigated using the methods of skin bioengineering. Good physical stability and applicative characteristics were obtained although LA strongly influenced the colloidal structure of the vehicle. LA per se and in APG‐based emulsion showed satisfying antioxidant activity that promotes it as mild multifunctional cosmeceutical efficient in the treatment and prevention of the photoaged skin. Employed assays were shown as suitable for the antioxidant activity evaluation of LA in APG‐based emulsions, but not for α‐tocopherol in the same vehicle.

Estimation of dermatological application of creams with St. John's Wort oil extracts.

Oleum Hyperici, the oil extract of St. John’s Wort (SJW), is one of the oldest folk remedies, traditionally used in the topical treatment of wounds, bruises, ulcers, cuts, burns, hemorrhoids and also as an antiseptic. Considering the advantageous characteristics of emulsion applications, in the present study we have formulated three O/W creams containing 15% (w/v) of SJW oil extract as an active ingredient. The aim was to estimate dermatological application of the prepared creams for the abovementioned indications. The extracts were prepared according to the prescriptions from traditional medicine, however with different vegetable oils used as an extractant, namely: Olive, palm and sunflower oil. The investigated O/W creams demonstrated significant antiinflammatory effects in an in vivo double-blind randomized study, using a sodium lauryl sulphate test. Both skin parameters assessed in the study (electrical capacitance and erythema index), were restored to the baseline value after a seven-day treatment with the tested creams. Almost all investigated SJW oil extracts and corresponding creams displayed the same antimicrobial activity against the most of the investigated microorganisms with obtained minimal inhibitory concentrations values of 1,280 µg/mL, 2,560 µg/mL or >2,560 µg/mL.

Hypericum Perforatum L. Hypericaceae/Guttiferae Sunflower, Olive and Palm Oil Еxtracts Attenuate Cold Restraint Stress – Induced Gastric Lesions

Three Hypericum perforatum L., Hypericaceae (St. John’s Wort) oil extracts (HPE) were prepared according to the prescriptions from traditional medicine – fresh flowering tops were macerated in three different vegetable oils: sunflower (E1), olive (E2) and palm oil (E3) for 40 days, exposed to the sunlight. The aim of the study was to investigate the gastroprotective activity of the obtained extracts in respect to their quercetin content. HPLC analysis confirmed the presence of quercetin in all of the investigated HPEs, but in different amounts: 15.1, 5.8 and 21.7 μg/mL in E1, E2, and E3, respectively. Gastroprotective activity was evaluated using cold-restraint stress (CRS) induced rat gastric mucosa lesions test. All of the HPEs showed gastroprotective activity, which was close to that achieved by the one of the most studied anti-ulcer flavonoids, quercetin [percentages of inhibition of ulcer index (UI) were 35, 62 and 40 % in E1, E2 and E3, respectively]. Contrary to the lowest quercetin content, HPE prepared with olive oil (E2) offered the highest protection against gastric damaging action of CRS. It may be assumed that this is due to other constituents of E2, which probably play an additional role in complex gastroprotective activity.

The estimation of the traditionally used yarrow (Achillea millefolium L. Asteraceae) oil extracts with anti-inflamatory potential in topical application

ETHNOPHARMACOLOGICAL RELEVANCE Olive or sunflower oil yarrow extracts have been extensively used against inflammatory disorders and skin wound healing in traditional medicine. AIM OF THE STUDY To evaluate oil yarrow extracts traditional use in treatment of topical/dermatological skin impairments, sodium lauryl sulfate test was applied, and in vivo measuring of the biophysical parameters (erythema index, skin capacitance and the pH of the skin) in the artificially irritated skin was performed. As traditionally olive and sunflower oil have been used equally for extracts production, the experiment was carried out to investigate whether any of the oil extractants has the advantage over the other, and if the method of extraction might influence the desired activity. The observed activity has been connected to the chemical profile of the investigated extracts and their antioxidative properties. MATERIALS AND METHODS In vivo measurements were performed using the appropriate probes for measuring skin capacitance, pH of the skin and erythema index (EI). The designed experiment enabled the evaluation of the anti-inflammatory effects of a seven-day application of oil yarrow extracts known in traditional medicine, on artificially irritated skin of volunteers. The chemical profile for the investigated samples was achieved applying the HPLC and UHPLC-MS methods. Also, ferric reducing antioxidant power (FRAP) test was performed to assess the antioxidant properties of the investigated samples. RESULTS The application of tested oil extracts on artificially irritated skin in vivo demonstrated the ability to re-establish their optimal pH and hydration of skin to the values measured prior to the irritation. Considering the EI transition, the investigated samples succeeded in re-establishing the baseline values, with no significant difference after three- and seven-day application. CONCLUSIONS The data obtained in the study showed that the oil yarrow extracts had an evident anti-inflammatory property. Namely, the investigated extracts demonstrated significant anti-inflammatory effect in an in vivo double blind randomized study, using a sodium lauryl sulfate test. The skin parameters assessed in the study (skin capacitance, pH and EI) were restored to the basal values after three- and seven-day treatment with the tested extracts. The shown effects were attributed to yarrow oil extracts composition. The yarrow oil extracts might be used as promising base in the phytopreparations designed for dermatological application as anti-inflammatory agents with a positive impact on the skin pH and its moisture content.

Supercritical carbon dioxide hops extractswith antimicrobial properties

Abstract Extracts obtained from hops (Humulus lupulus L., Cannabaceae) by supercritical fluid extraction (SFE), SFE followed by isomerization, as well as by conventional technique, were investigated for their chemical composition and antibacterial activity against selected foodborne pathogens and microorganisms capable to cause the food spoilage. The antibacterial activity of the extracts was compared with the antibacterial activity of xanthohumol, compound known for its broad pharmacological properties, isolated from the raw material remained after the SFE. Xanthohumol (XH, 96%) proved to posses the most prominent activity against all the tested strains, with the MIC values ranged between 2.5 and 20 μg mL-1. Supercritical hops extract and potassium isomerized supercritical hops extract showed strong antibacterial activity against the tested strains as well. Escherichia coli was not affected by the extracts, meaning that their oral admission would not cause the same problem as antibiotic application in intestinal flora. The chemical composition of the investigated hops extracts was analysed by GC-MS. Contents of α-acids, β-acids, iso-α-acids and xanthohumol in the samples were determined by HPLC. Graphical Abstract

Antioxidative, membrane protective and antiapoptotic effects of melatonin, in silico study of physico-chemical profile and efficiency of nanoliposome delivery compared to betaine

The mechanisms through which melatonin exerts its inhibitory effect on cell death remains insufficiently clarified. The subject of the present study is the evaluation of the hepatoprotective effects of melatonin on the inhibition of apoptotic and oxidative processes and activation of survival pathways, in comparison with betaine, in a primary culture of hepatocytes that have undergone Fas-ligand apoptosis. Melatonin exerted a protective effect on membrane bilayer stability through the inhibition of phosphatidylserine externalization, Bax expression and xanthine oxidase catalyzed free-radical liberation. It also reduced liberation of cellular and membrane-bound enzymes and endonuclease catalyzed DNA fragmentation. Betaine hydrochloride did not exert these effects, when administered alone or co-incubated with anti-Fas antibodies. The in silico Molinspiration tool, which was employed to calculate melatonin and betaine physico-chemical properties and membrane interaction indicated that melatonin may easily cross and interact with biological membranes and maintain membrane phospholipid structural topography. It was documented to occur via non-receptor mechanisms in more than 75%; this may clarify melatonin as suitable for nanoliposome-based delivery, where it was also able to successfully counteract induced oxidative stress. This could not be considered for betaine hydrochloride. The use of lipophilic compounds like melatonin, encapsulated in nanoliposomes, could therefore be a preferable tool in the successful membrane-preservation therapy of liver apoptosis, rather than the use of hydrophilic compounds, like betaine hydrochloride.