Search

This study discusses the possibility of supercritical solvent impregnation of polypropylene and polyamide fabrics as well as cellulose acetate in the form of beads and films with pyrethrum extract in order to fabricate functionalized materials with repellent properties. Impregnation experiments were performed at temperature of 40 degrees C and pressures of 10 and 20 MPa in supercritical carbon dioxide. The time of impregnation was varied from 0.5 to 18 h. Loadings of pyrethrum extract into polypropylene fabrics and cellulose acetate films higher than 30% were attainable at 10 MPa. In the case of polyamide fabrics and cellulose acetate beads, the pressure of 20 MPa was needed for obtaining the loadings higher than 11%. Pyrethrum extract release study revealed different capabilities of the tested materials for the repellent release, implying a variety of their possible applications. The impregnated solids were characterized by SEM, DSC, FTIR, Mercury intrusion porosimetry analyses and HPLC analyses. This the peer-reviewed version of the article: Pajnik J. et al., Journal of Supercritical Fluids, 2018, 136, 70-81, doi: [https://doi.org/10.1016/j.supflu.2018.02.014] Published version: [https://cer.ihtm.bg.ac.rs/handle/123456789/2298]

Lichens represent a promising source of antimicrobial, cytotoxic and antioxidant agents. Their great pharmacological potential lies in the fact that they represent specific symbiotic organisms and thus possess natural roles allowing them to be highly adaptable to different environmental conditions. On the other hand, stated biological activities of lichens with prospective medicinal significance may be connected to their long-term use in traditional treatment of various ailments. Genus Usnea from the Parmeliaceae family is certainly one of the best studied in terms of chemical composition and biological properties of its extracts and/or isolated compounds. In the first part of the study, a detailed review of literature has been performed yielding a detailed report on the investigations of biological activities of the lichens belonging to this genus. In the second part of the study, chemical composition of the lichens from the genus was described and, additionally, a survey of the biological properties of the most representative secondary metabolites in these lichens has been reported. It could be concluded that the extracts and/or isolated compounds from the lichens belonging to the genus Usnea may be considered a valuable source of prospective drug candidates with potential clinical relevance.

A comprehensive study on essential oil and different solvent extracts of Sideritis romana L. subsp. purpurea (Tal. ex Benth.) Heywood (Lamiaceae) from Montenegro is reported. The gas chromatography-mass spectrometry analysis of the essential oil revealed a total of 43 components with bicyclogermacrene (23.8%), germacrene D (8%), (E)-caryophyllene (7.9%) and spathulenol (5.5%) as the major ones. Sesquiterpenoid group was found to be the most dominant one (64.8%), with 19.9% of the oxygenated forms. In the crude methanol extract of the investigated plant, obtained by Sohhlet exraction, the total phenol content was 14.7 ± 0.4 mg of GA/g, the total flavonoids were 0.29 ± 0.03% expressed as hyperoside percentage, whereas the total tannins content was 0.22 ± 0.04% expressed as pyrogallol percentage. For the antimicrobial activity determination, the following microorganisms have been used: methicillin-susceptible Staphylococcus aureus (MSSA (American Type Culture Collection (ATCC) 29213)) and methicillin-resistant S. aureus (MRSA (clinical strain)), Escherichia coli (ATCC 25922), carbapenem-susceptible Klebsiella pneumoniae (clinical strain), carbapenem-resistant K. pneumoniae (clinical strain) and Candida albicans (ATCC 14053). The essential oil showed high potency against MSSA and MRSA, both at high (~5 × 10⁵ CFU/mL) and low (~5 × 10³ CFU/mL) inoculum. With respect to MSSA, the minimal inhibitory concentration (MIC) value was 0.307 mg/mL, with bactericidal activity obtained at 0.615 mg/mL, while, in the case of MRSA, the MIC and minimal bactericidal concentration (MBC) values were 0.076 and 0.153 mg/mL, respectively. Regarding anti-Candida albicans activity, the MIC value was 2.46 mg/mL without reaching fungicidal activity. In addition to the observed essential oil efficacy, different solvent extracts were analyzed for their antimicrobial activity. Similarly to the essential oil, thehighest efficacy was observed against both MSSA and MRSA strains, at high and low inoculums, in the case of the 1,2-dichloroethane and methanol extracts. A potent fungicidal activity has been also found for the n-hexane and 1,2-dichloroethane extracts. It can be concluded that Sideritis romana L. subsp. purpurea (Tal. ex Benth.) Heywood provides a wide range of application in different fields such as phytochemistry, pharmacology, toxicology or pharmacognosy.

The antioxidant and antibacterial properties of Greek oregano extracts obtained by fractional supercritical fluid extraction (SFE) with carbon dioxide were investigated and compared with the properties of essential oil obtained by hydrodistillation. According to DPPH., hydroxyl radical and superoxide anion radical scavenging activity assays, the supercritical extracts expressed stronger antioxidant activity comparing to the essential oil. The most effective was the supercritical extract obtained by fractional extraction at 30 MPa and 100 degrees C after the volatile fraction had been extracted at lower pressure. At the same time this extract showed strong antibacterial activity against staphylococci, including MASA strain, but did not affect Escherichia coli of normal intestinal flora. The essential oil obtained by hydrodistillation showed stronger antibacterial activity against E. coli, Salmonella and Klebsiella pneumoniae, comparing to the supercritical extracts but at the same affected the normal gut flora.

Ethnopharmacological relevance Alchemilla vulgaris is an important remedy in European folk medicine, known for its astringent and anti-inflammatory properties; it is traditionally used to heal gynecological and gastrointestinal diseases. Despite its folkloric use in wound healing, there is a lack of scientific data to support this therapeutic application. Aim of the study To analyze the wound healing potential of different solvent A. vulgaris extracts per se and after incorporation into hydrogels as topical vehicles, using two complementary methods - in vitro wound healing assay with L929 fibroblasts and in vivo assessment of skin barrier repair potential. Besides scientific justification of the traditional usage, we aimed to ephasize the importance of a proper vehicle for herbal extracts. The wound healing activity has been connected to the chemical profile of the investigated extracts, their antioxidative properties, but also to pH of the investigated gels and their mechanical characteristics. Materials and methods Antioxidant activity of investigated extracts was estimated using both 2,2-diphenyl-1-picrylhydrazyl and β-carotene/linoleic acid models. Chemical profile was achieved applying spectrophotometric and HPLC methods. In vitro scratch assay with L929 fibroblasts, and in vivo study of skin barrier repair potential of hydrogels with A. vulgaris extracts on human skin employing biophysical measurements, were performed in order to confirm the wound healing potential of A. vulgaris. Texture analysis of the gels was performed alongside the pH measurements. Results All tested extracts and gels accelerated the wound healing process while the effect of ethanolic extract on migration of fibroblasts was the most pronounced. The highest extent of wound closure was also observed for the ethanolic extract. The most favorable effect on in vitro wound healing was observed for gel with propyleneglycolic extract. Results of in vivo study were in line with in vitro findings. Healing potential may be attributed to phenolic compounds found in A. vulgaris extracts, low pH of the gels, and the satisfying antioxidant activity of the extracts. Parameters obtained by textural analysis indicated satisfying mechanical properties of the gels, relevant to topical application. Conclusion Our study offers pharmacological evidence on the folkloric use of A. vulgaris in wound treatment, particularly after incorporation into hydrogel, and underlines an importance of a proper vehicle for incorporation of herbal extracts intended for topical treatment.

There is a growing research interest on investigation of appropriate alternatives of natural origin to synthetic additives used in food products. The aim of this study was to investigate correlation of chemical composition of the best known biomarker plants from the locality of Vrujci Spa, Serbia with their antioxidant activity in vitro, in terms of their potential usage as feedstock materials in the food industry. Total phenolic (TP), total tannins (TT) and total flavonoid (TF) contents were determined using spectrophotometric methods. Qualitative and quantitative analyses of major phenolics by high-performance liquid chromatography (HPLC) were also used. Major phenolic acids identified in analyzed species were chlorogenic, caffeic and ferulic acid, while predominant flavonoids were flavonols (hyperoside, rutin, isoquercitrin, quercetin) and, in lesser extent flavones (luteolin, apigenin and their 7-O-glucosides). Vitexin 2-O″-rhamnoside was found only in Crataegus species. Antioxidant capacity was estimated by the following methods: 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) test and ferric reducing antioxidant power (FRAP). Majority of investigated plants had high levels of phenolics and exhibited noteworthy antioxidant activity. Moreover, a positive linear correlation was found between TP and TT with antioxidant capacities regarding both methods used. The plants showing the highest antioxidant activities were Veronica officinalis, Mentha pulegium and Fragaria vesca. Hypericum perforatum demonstrated by far the most potent antioxidant activity, as well as TP and TT content. Our results indicate that, in addition to their traditional usage in folk medicine, these plants represent a valuable source of natural antioxidants, and thus may be considered as great potential for the food industry, representing possible alternatives to synthetic additives.

A study on essential oil fractions of the Western Balkan endemic Sideritis romana L. subsp. purpurea (Tal. ex Benth.) Heywood collected in Montenegro is reported. The 24-h systematic steam distillation extraction procedure was performed. The gas chromatographic/mass spectrometric (GC/MS) analysis of the fractions showed γ-elemene and spathulenol as two main constituents, revealing a new chemotype of this plant species. Although varying in the content of these two main compounds, which makes the fractions quite different between each other, evaluation of the anti-Candida activity showed the lack of any significant efficacy.

Introduction. Rational phytotherapy is a modern concept of using plant-originated drugs which has emerged from the need to improve phytotherapy in order to make the use of herbal remedies more efficient and safer. The aim of this study was to give the health-care workers more information on the manufacturing process of high quality phytopreparation following principles of Good Manufacturing Practice and Good Laboratory Practice on the example of herbal sedative, Odoval S? capsules. Material and Methods. This study was designed to reflect the production process of a high-quality and safe herbal remedy, starting from defining the formulation and the production procedure to the quality control of raw materials, characterization of the final product, and testing stability of active ingredients in the capsules. Results. Formulation of the phytopreparation, validation of the production process, quality control and stability testing, all together have resulted in the production of capsules with defined valeric acid content (1 mg valeric acid per capsule). Discussion. The preparation is recommended to relieve the symptoms caused by chronic stress (anxiety, irritability, fatigue, lack of concentration, heart palpitations) and for mild insomnia. Conclusion. This paper presents the complete cycle of the production of a phytopreparation on the example of a new herbal sedative - Odoval S? capsules.

Helichrysum italicum presents a valuable source of natural bioactive compounds. In this work, a literature review of terpenes, phenolic compounds, and other less common phytochemicals from H. italicum with regard to application of different separation methods is presented. Data including extraction/separation methods and experimental conditions applied, obtained yields, number of identified compounds, content of different compound groups, and analytical techniques applied are shown as corresponding tables. Numerous biological activities of both isolates and individual compounds are emphasized. In addition, the data reported are discussed, and the directions for further investigations are proposed.

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.

Oleum Hyperici, the oil extract of St. Johns 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 mu g/mL, 2,560 mu g/mL or >2,560 mu g/mL.

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.

From a botanical perspective, the large number of species of the genus Sideritis coupled with the tendency to hybridise between the different species has prompted an accurate study designed to clarify all the controversial points in the botanical classification of this genus. The past phytochemical studies of the Sideritis species have been performed with different extracts from the aerial parts, essential oils and isolated compounds such as diterpenoids, flavonoids or phenylpropanoid glycosides. These investigations have justified the traditional uses for these plants and provided direction for new pharmacological research. In recent years, other compounds such as iridoids, coumarins, lignans and phenylpropanoid glycosides have also been isolated and identified. The results have shown that essential oils act as good antimicrobial agents against both Gram-positive and Gram-negative bacteria and against the Candida albicans fungus. Diterpenoids have shown antimicrobial, anti-inflammatory and antifeedant activity, and flavonoids are active as anti-ulcerous, anti-inflammatory and antioxidant agents. Future research should focus on the pharmacological activity of these isolated compounds to find new active principles and identify their mechanisms of action. In addition, it would be interesting to investigate new pharmacological activities apart from those used in traditional medicine; recent studies have shown that diterpenes and some diterpene derivatives act as effective anti-HIV and antiproliferative agents.

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.

Some medicinal features of olive leaf have been known for centuries. It has been traditionally used as an antimicrobial and to prevent and treat diabetes mellitus and heart disease. Whether olive leaf, a natural antioxidant, influences the gastric defense mechanism and exhibits gastroprotection against experimentally-induced gastric lesions remains unknown. In this study, the content of total phenols, total flavonoids and tannins in olive leaf extract (OLE) were determined. Seven phenolic compounds were identified and quantified (oleuropein, caffeic acid, luteolin, luteolin-7-O-glucoside, apigenin-7-O-glucoside, quercetin, and chryseriol). Furthermore, the protective activity of the OLE in gastric mucosal injury induced by a corrosive concentration of ethanol was investigated. In relation to the control group, pretreatment with OLE (40, 80 and 120 mg kg-1) significantly (p < 0.001) attenuated the gastric lesions induced by absolute ethanol. The protective effect of the OLE was similar to that obtained with a reference drug, ranitidine. The results obtained indicate that OLE possesses significant gastroprotective activity, and that the presence of compounds with antioxidative properties would probably explain this effect.

The aim of the present work was to examine if hydrogen peroxide treatment of sugar beet fibre that aimed at improving its physicochemical properties would impair its antioxidant potential. Three different sugar beet fibres were obtained from sugar beet - non-treated fibre (NTF) from sugar beet cossettes extracted with sulphurous acid, treated fibre (TF) from NTF treated with hydrogen peroxide in alkaline solution and commercially available Fibrex(®) . The antioxidant activity of extractable and non-extractable fibre fractions in ethanol/water mixture (80:20, v/v) of three fibre samples was estimated. Non-extractable fractions obtained after alkaline treatment of investigated fibres were much higher in phenolic compounds and possessed higher antioxidant potential than extractable fractions. Ferulic acid was proven to be the dominant phenolic acid. Regarding both extractable and non-extractable fractions, Fibrex(®) had the highest antioxidant activity in chemical tests, while NTF was superior in comparison with TF. Based on the results of Caco-2 cells-based test, all non-extractable fractions possessed potential for reactive oxygen species inhibition. Regarding the extractable fractions, only the TF manifested this effect.Copyright © 2016 John WileySons, Ltd.

Supercritical CO2 (SC-CO2) extraction of fenugreek seeds (Trigonella foenum-graecum L, Fabaceae) was analyzed using different pressure and temperature (16.6-33.4 MPa and, 30-50 degrees C), which corresponded to SC-CO2 density between 779 and 957 kg/m(3). The yield of extract as well as its composition were determined and analyzed for different solvent consumption defined as mass of used CO2 for extraction per mass of dry fenugreek seeds. A central composite design (CCD) combined with response surface methodology (RSM) was used to study the best extraction conditions which determined that 27.0 MPa, 42.6 degrees C, and 24.6 gCO(2)/g(dm), were the optimal conditions for isolation of total extract. The calculated yield of extract at these conditions defined by SC-CO2 density of 890 kg/m3 could be 2.57% based on fenugreek seed. Moreover, the best extraction conditions for obtaining extract which contains the maximal amount of sterols, or maximal amount of vitamin E or vitamin D were also analyzed. These RSM optimizations showed that at slightly lower pressure (22.8-24.8 MPa), temperature (39.9-42.0 degrees C) and consumption of SC-CO2 (19.0-19.7 gCO(2)/g(dm)) is followed by smaller amount of isolated extract but with maximum content of sterols (16.28%), or vitamin E or vitamin D (1.44% and 0.71%, respectively) at these conditions. Calculation of molar solubility of total extract, and apparent solubility of sterols, vitamin E and D in SC-CO2 was performed at different pressure and temperature. They were correlated to density of SC-CO2, temperature at which extractions were performed and Hildebrand solubility parameter of solvent and solubility parameter of compounds which are the main constituents of extract. These calculations also show that the highest molar solubility of total extract and apparent molar solubility of its constituents could be obtained using optimal density of SC-CO2 (830-890 kg/m(3)) defined by corresponding values of temperature and pressure (310-320 K and 20-25 MPa).

There is a worldwide ongoing investigation for novel natural constituents with cytotoxic and antioxidant properties. The aim of this study was to investigate chemical profile and stated biological activities of the supercritical CO2 extract (SCE) of old man's beard compared to the extracts obtained using the conventional techniques (Soxhlet extracts and macerate). The most abundant compound identified was usnic acid, which content was inversely proportional to the polarity of the solvent used and was the highest in the SCE, which was the sample revealing the highest cytotoxic activity in tested tumor cell lines (B16 mouse melanoma and C6 rat glioma), with lower IC50 values compared to pure usnic acid. Further investigations suggested both SCE and usnic acid to induce apoptosis and/ or autophagy in B16 and C6, indicating higher cytotoxicity of SCE to be related to the higher degree of ROS production. A good correlation of usnic acid content in the extracts and their antioxidant capacity was established, extricating SCE as the most active one. Presented results support further investigations of SCE of old man's beard as a prospective therapeutic agent with potential relevance in the treatment of cancer and/ or in oxidative stress-mediated conditions.

Topical treatment of skin infections is often limited by drawbacks related to both antimicrobial agents and their vehicles. In addition, considering the growing promotion of natural therapeutic products, our objective was to develop and evaluate naturally-based emulsion system, as prospective topical formulation for skin infections-treatment. Therefore, alkyl polyglucoside surfactants were used for stabilization of a vehicle serving as potential carrier for supercritical CO2-extract of Usnea barbata, lichen with well-documented antimicrobial activity, incorporated using two protocols and three concentrations. Comprehensive physicochemical characterization suggested possible involvement of extract's particles in stabilization of the investigated system. Raman spectral imaging served as the key method in disclosing extract's particles potential to participate in the microstructure of the tested emulsion system via three mechanisms: (1) particle-particle aggregation, (2) adsorption at the oil-water interface and (3) hydrophobic particle-surfactant interactions. Stated extract-vehicle interaction proved to be correlated to the preparation procedure and extract concentration on one hand and to affect the physicochemical and biopharmaceutical features of investigated system, on the other hand. Thereafter, formulation with the best preliminary stability and liberation profile was selected for further efficiency and in vivo skin irritation potential evaluation, implying pertinent in vitro antimicrobial activity against G+ bacteria and overall satisfying preliminary safety profile.

Chaerophyllum coloratum is an endemic plant for the Western Balkans. Despite of its common occurrence in the Sub-Mediterranean part of Dinaric countries, knowledge about its structure and biologically active compounds is rather poor. The plant material was collected in Montenegro. The structure was studied on paraffin sections, as well as on native sections stained with standard reagents for histochemical analysis. Chemical analyses of the essential oils from its leaves, flowers, roots, stems and fruits were performed. The total phenolic, flavonoid and tannin contents in different plant organs were determined, as well, in order to give an insight in the pattern of secondary metabolites distribution. Although it was shown that morphologically and ecologically Chaerophyllum coloratum differed significantly from other species of the genus native in the flora of the Western Balkans, it shared common structural features of the genus. In a comparative analysis of 33 anatomical characters of C coloratum and six previously studied Chaerophyllum species, C coloratum clustered with Chaerophyllum aureum. In addition it was closely linked with two other species of Chrysocarpum section: Chaerophyllum bulbosum and Chaerophyllum aromaticum. Histochemical and chemical analysis of secondary metabolites showed the presence of essential oil in all studied plant organs, but differing both in composition and content of individual compounds. The essential oil from the roots is characterized by a high content of monoterpene myrcene (72.18%). In the case of stem monoterpens beta-(E)-ocimene (33.59%), beta-(Z)-ocimene (20.43%) and terpinolene (10.77%) were predominant compounds, while the principal constituents in the oil of leaves were identified as spathulenol (10.19%), p-cymene-8-ol (9.45%) and p-cymene (7.6%). Monoterpenes terpinolene (17.08%), p-(Z)-(E)-ocimene (6.96%) were identified as the main constituents of the flower oil, while in the fruits, the constituents with the highest amount were caryophyllene oxide (6.93%), (Z)-beta-farnesene (5.92%), (E)-pinocarveol (5.56%) and myrtenol (5.25%).

Genome editing tools, such as TALEN (transcription activator-like effector nuclease) or CRISPR-Cas9 (CRISPR-associated protein-9 nuclease) systems, enable functional studies by targeted gene knockout. They introduce double-stranded breaks (DSBs) into a DNA molecule in a sequence-specific manner, thereby stimulating the error-prone non-homologous end joining repair mechanism, leading to probable gene inactivation when the coding sequence is targeted. Vectors for expression of TALEN and Cas9-based constructs targeting the human IL6ST and HNF1A genes were assembled and tested for their ability to introduce DSBs when transfected into cultured cells using the luciferase assay. The Cas9-based construct targeting the IL6ST gene was shown to be active, while the two TALEN-based constructs did not introduce DSBs above background level. Both the TALEN and the CRISPR-Cas9 constructs targeting the HNF1A gene were found to be active, with the TALEN showing higher activity in a dose-dependent manner. The constructed genome-editing tools can be used for functional analysis of the putative role of HNF1A and IL6ST genes in IgG glycosylation, as shown previously by genome wide association studies. This work is licensed under a Creative Commons Attribution 4.0 International License.

Analysis of yield and composition profile of Melissa officinalis (Lemon balm) extract obtained by two-step CO2 extraction at high pressure was analyzed. The first or essential oil fraction, collected at 10 MPa and 40 degrees C, contained mostly the high volatile and aromatic compounds. The second fraction was consenquently extracted at 30 MPa using different temperature: 25 degrees C (higher pressure than critical pressure but at temperature below the critical; HPCP-CO2), 40 degrees C or 100 degrees C (supercritical CO2; SC-CO2). Obtained yield of essential oil fraction was 0.45% at 10 MPa, while depending on applied extraction temperature (HPCP-CO2 or SC-CO2), the yields of the second fractions collected at 30 MPa varied from 0.44% to 0.94%. Within the study, the content of active substances in obtained extracts was analyzed, too. For comparison with CO2 extracts (HPCP or SC) the yield and characterization of essential oil obtained by hydrodistillation was accomplished, as well. A significant difference was noted not only between the essential oil isolated by hydrodistillation and SC-CO2 extraction (40 degrees C, 10 MPa) but also between total extracts obtained by HPCP-CO2 and SC-CO2 extraction. The pseudo kinetic model was applied to define SC-CO2 extraction of Lemon balm to describe the relation between the yield of extraction and CO2 consumption. Antioxidant activity of obtained extracts was determined by free radical scavenging capacity (RSC) and significant correlation of antioxidant capacity and composition profile between obtained extracts was observed.

Ethanol extracts of the aerial parts of six Hypericum species, H. barbatum, H. androsaemum, H. richerii, H. hirsutum, H. perforatum and H. perforatum cultivated on mountain Tara were tested for antiinflammatory activity in comparison with indomethacin (IND) using the carrageenan-induced rat paw edema test. It was found that all examined extracts produced antiinflammatory activity, particularly those from H. hirsutum, and both wild and cultivated H. perforatum. A dose-dependent antiinflammatory effect was especially pronounced in extracts showing some lower antiinflammatory activity. There was no correlation between the amount of hypericin in the extracts and their antiinflammatory activity.

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.

The aim of this study was to investigate the physicochemical properties of peloid, its mineralogical composition and its antimicrobial activity, including the presence of algae, with the aim of considering its dermo-cosmetic application, such as anti-skin aging and treatment. Physicochemical analysis showed that peloid from Ulcinj coast, contains minerals, necessary for smooth performance of skin functions, as well as, the whole body. A studied peloid sample showed significant antimicrobial activity of Candida albicans strain, and the presence of algae of Bacillariophyta division, known to have a beneficial effect on skin health. Results of examination of peloid from Ulcinj locality, recommended it as a high quality natural substance applicative in dermo-cosmetic preparations for treatment of problematic skin.

For the first time, an ultrasound-assisted extraction was used to investigate isolation of anthocyanin-containing extracts from the blackberry cultivar “Cacanska Bestrna” widely grown in Serbia. Sonication time and temperature influence on the yield, active substances content and antioxidant activity of the extracts isolated from blackberry fruit puree was discussed. Ultrasound-assisted extraction with acidified ethanol at room and moderately high temperature (40 °C) enabled rapid isolation (15–30 min) of anthocyanin-containing blackberry extracts with the high yields (5.3–6.3%). Both increase of the sonication temperature and time within studied ranges had positive effect on the extraction yield. The HPLC analysis revealed the presence of the anthocyanins in the investigated samples. After hydrolysis of all the samples, aglycone–cyanidin was detected in the samples. Determined cyanidin content in the obtained blackberry extracts (0.7–1.0%) was up to 20 times higher than in the blackberry juice. Cyanidin content was positively affected by sonication time and temperature increase. The antioxidant properties of the extracts were analyzed by the ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity assays. Positive effect of increase of the sonication time and/or temperature on the scavenging activity of the extracts against free DPPH radicals and thus lower IC50 values was demonstrated. Strong linear correlation of DPPH radical scavenging capacities of the extracts with content of cyanidin and tannins was established. FRAP values significantly correlated with total tannin content in the extracts. The analyses of composition and antioxidant properties in this study revealed great potential of blackberry ultrasonic extracts of the cultivar “Cacanska Bestrna” for further use in food and pharmaceutical industries.

Sideritis scardica (mountain tea) is an endemic plant on the Balkan Peninsula traditionally used for treating different conditions, mainly of inflammatory nature. This study was aimed to examine the cytotoxic activity of different S. scardica extracts against the rat glioma C6 line and rat astrocytes in primary culture. The obtained data revealed that diethyl ether (extract 2) and ethyl acetate (extract 3) extracts of S. scardica exerted a cytotoxic effect on C6 rat glioma cells. Diethyl ether extract induced an increase in reactive oxygen species production, leading to apoptotic and autophagic cell death. Ethyl acetate extract induced G 2 M cell cycle arrest and autophagy. None of the tested extracts was cytotoxic to rat astrocytes in primary culture. Cytotoxic effects of S. scardica extracts were, at least in part, mediated by their flavonoid constituents apigenin and luteolin that, when applied alone, induced cell cycle arrest, apoptosis, and autophagy.

SUMMARY In this study we investigated the effects of four emollients (cetearyl octanoate, decyl oleate, C12-15 alkyl benzoate and olive oil) incorporated at a concentration of 10% on the structural properties of the cosmetic oil/water (o/w) creams (creams K1-K4, respectively) as well as the in vivo effects of those creams on pH and hydration level of the healthy volunteer’s skin after a single application. Creams’ structure was investigated throughout polarization microscopy. Anisotropic structures were found, i.e. stabilization of creams’ structure was performed by forming lamellar gel-crystalline and liquid-crystalline phase, different for different creams, which was probably caused by the presence of different emollients. The effects of creams (K1-K4) on skin hydration level and pH (30, 90, 150 and 300min after single application) were examined using Multi Probe Adapter (MPA ® 9, Courage& Khazaka Electronic, Germany), with appropriate sonde, Corneometer ® CM 825 for measuring capacitance as a measure of skin moisture level, while the skin pH was measured using Skin-pH-Meter, PH 900 sonde.The results indicate that the presence of the different emollients in o/w creams (10%) led to the creams'different effects on the skin hydration level. Cream with decyl oleate leads to a good initial hydration effect and probably rapid release of large amounts of water from the cream. Creams containing both C12-C15 alkyl benzoate or olive oil had a weaker initial effect on the skin moisture, showing good occlusive properties. A cream containing 10% cetearyl octanoate showed the lowest skin hydration potential.

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.

Background Osteoporosis is the most common metabolic bone disease. Postmenopausal loss of estrogen is a major risk factor. Objectives To assess if total ethanolic extract of Sideritis scardica could prevent osteoporosis in ovariectomized female rats. Methods Female Wistar rats were divided in three groups: I-sham operated (control), II-ovariectomized-OV and III-ovariectomized, treated orally with S.scardica extract (300 mg/kg) during 20 weeks-OVT. In 18th experimental week oral glucose tolerance test has been performed, as well measurement of bone mineral density-BMD. After killing, femurs, plasma and erytrocytes were isolated. The following parameters were measured: morfometric bone parameters, wet and dry bone mass, biomechanical testig (three point flexural test), Ca and P in urine and plasma, alkaline phosphatase in plasma, malondialdehyde, catalase activity, reduced gluthation (GSH), total SH groups, free fatty acids and fructozamines. Results There was no significant difference in Ca and P concentration in plasma between groups, but urinary Ca and P were significantly higher in OV rats (Ca 0.67±0.24 mM, P 3.29±0.77 mM) in comparison to control (Ca 0.434±0.21 mM, P 2.32±1.00 mM) and OVT group (Ca 0.52±0.19 mM, P 2.89±0.42 mM) (p 2 , for OV group 2726.68±93.4 N/m 2 and for OVT group 3257.2±86.5 N/m 2 (p Conclusions S. scardica extraxct prevents bone loss in ovariectomized rats, increases bone resistance to mechanical load and reduces cardiovascular metabolic risc factors associated with menopause. Disclosure of Interest None declared