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Mastitis is one of the most common and costly diseases affecting dairy cows worldwide. Since antibiotic resistance has become a global threat to both animal and human health, it is becoming more urgent to continuously search for new therapeutical alternatives for the control and treatment of bovine mastitis. Hence, our research aimed to test the therapeutic use of two essential oils (EOs) based on their chemical composition, antibacterial and antioxidant potential. The present study was conducted by collecting milk samples from the cows diagnosed with clinical or subclinical mastitis with the aim of isolating and identifying bacterial strains. The antioxidant potential of essential oils of Menthae piperitae (MP) and Melissa officinalis (MO) was evaluated in several in vitro assays. In the MP EO, a total of 38 compounds were identified, with menthol as the dominant compound, whereas in MO EO 51 compounds were identified. Furthermore, the values of minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) have been used to quantitatively measure the antibacterial activity of each essential oil. In accordance with which, MP EO samples exhibited a higher degree of antibacterial activity than MO EO. Thus, EOs have been shown to be promising alternatives to antibiotics because of their availability, biodegradability, and lower risk of side effects as compared with conventional, antimicrobial treatment. Nevertheless, further clinical studies are needed to test the potential role of EOs in treating mastitis in dairy cows.

Rosmarinic acid is a phenolic compound commonly found in the Lamiaceae (Labiateae) plant species. It is considered responsible for a wide spectrum of biological and pharmacological activities of plants containing this compound. The aim of the current review is to present the fate of rosmarinic acid inside the human body, explained through pharmacokinetic steps and to briefly present the health benefits of RA. Pharmacokinetics was at first studied in animal models, but several studies were conducted in humans as well. This compound can be applied topically, pulmonary, intranasally, and via intravenous infusion. However, peroral application is the main route of entry into the human body. Presumably, it is mainly metabolized by the gut microflora, providing simple, more easily absorbed phenolic units. Inside the body, the rosmarinic acid molecule undergoes structural changes, as well as conjugation reactions. Renal excretion represents the main path of elimination. Previously conducted studies reported no serious adverse effects of herbal remedies containing RA, as well as their positive effects on human health. In addition to in vitro studies, clinical investigations suggested its benefits in dermatological, allergic, and osteoarthritic disorders, as well as for improving cognitive performance and in metabolic syndrome treatment. Future studies should investigate the kinetics during long-term application in patients who would have potential benefits from RA usage. Pharmaceutical formulations designed to prevent the fast metabolism of RA and allow its penetration into other compartments of the human body are also interesting topics for future research.

St. John’s wort has been used for centuries in traditional medicine of many cultures, and nowadays it is well-known as a clinically important antidepressant drug. Considering the rising market demand for Hyperici herba, quality control of crude drug is of paramount importance. In this paper we performed HPLC-DAD chemical profiling of St. John’s wort tea samples obtained at local markets, pharmacies and health food stores in the Balkan Peninsula countries, Austria and Turkey. Furthermore, water alcoholic extracts of the collected samples were evaluated in terms of their antioxidant potential, as well as the ability to inhibit biologically important enzymes such as acetylcholinesterase, monoamine oxidases A and B (MAO-A and MAO-B), α-amylase and α-glucosidase. Significant variability within the samples in the quantities of hypericin, hyperforin, rutin, quercetin, gallic, chlorogenic, caffeic and p-hydroxybenzoic acid was noticed. Chemotaxonomic modelling enabled the identification of three clusters of samples based on the quantities of rutin, hypericin and hyperforin. Generally, the extracts exhibited a significant potential to inhibit MAO-A (median IC50 = 10.01 μg/mL) and α-glucosidase (median IC50 = 12.40 μg/mL). The results of antioxidant potential evaluation suggest strong neutralization of hydroxyl and nitroso radicals, but moderate inhibition of lipid peroxidation process. Overall, the conducted study emphasizes the possibility of St. John’s wort teas quality control, taking into account unavoidable variability of chemical profile as well. The stated is of great importance when grinding degree of herbal material excludes taxonomic identification of biological origin as an option within quality control.

Traditional medicinal plants are used as an additional form of type 2 diabetes therapy in which oxidative stress is one of the progression factors. The aim of the study was the evaluation of antihyperglycemic and antioxidant potential of tea mixture registered as food and marketed for treatment of type 2 diabetes mellitus. Chemical profiling of the extracts included quantification of total phenolics and flavonoids, while antihyperglycemic potential was estimated through determination of inhibition potential of α-amylase and α-glucosidase. Additionally, extracts were characterized for specific phenolic compounds by high performance liquid chromatography (HPLC). The in vitro antioxidant activity was estimated by application of DPPH, OH, NO and FRAP tests. The greatest anti-α-amylase potential is attributed to the Phaseoli legumen, while the highest potential to inhibit α-glucosidase was recorded for Olivae folium. Generally, screening of the phenolic compounds enabled us to conclude that a high level of correlation exists between anti-α-amylase potential and the quantified amounts of total phenolics, as well as gallic acid, while the chlorogenic acid is most probably responsible for the inhibition of α-glucosidase activity. Therefore, the high total ratio (40%, m/m) of Olivae folium and Phaseoli legumen in the evaluated tea mixture can be stated as fully justified. However, future in vivo research is required for further understanding of the antihyperglycemic activity mechanisms of secondary metabolites found in specified herbal components, as well as to evaluate their synergistic effect when applied in a form of tea mixture.

It is considered that different plant organic compounds, known as phytoncides are creditable for positive effects of Forest therapy on human health. Ongoing pandemic has also put these compounds in research focus considering potential use for novel corona virus treatment. This research was conducted in order to examine the potential of Tilia cordata Mill. considering volatile organic compounds (VOCs) content for the use in Forest therapy. In order to determine qualitative and quantitative content of phytoncides, as well the variability among genotypes growing in the same environmental conditions, leaf samples from genotypes of T. cordata were collected from the Fruška gora (Serbia) during 2019. VOCs determined in fresh herbal material were analyzed by HeadspaceGC/MS and VOCs determined in essential oil were obtained by GC/MS. The results showed the presence of 17 different phytoncides (monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpene hydrocarbons, oxygenated sesquiterpenes, aliphatic compounds and diterpenes) in all tested genotypes. According to the mean values of the obtained results, the most abundant classes of phytoncides among the examined genotypes were monoterpene hydrocarbons (87.05), followed by aliphatic compounds (36.59) and oxygenated sesquiterpenes (34.60). The highest coefficient of variation (CV) among phytoncide content in tested T. cordata genotypes has been observed within oxygenated monoterpenes (43.12%), sesquiterpene hydrocarbons (28.18%), and diterpenes (27.04%). Some individual monoterpene hydrocarbons, such as o-cymene (19.92%) and a-pinene (17.40%) had the highest CV in terms of its presence in analyzed genotypes. In addition, the principal component analysis (PCA) showed more notable clustering within the VOCs determined in essential oil in comparison to VOCs determined in fresh herbal material. Considering the phytoncide content detected in T. cordata leaves, this species has high potential for the use in human health improvement within Forest therapy.

As proper wound management is crucial to reducing morbidity and improving quality of life, this study evaluated for the first time the wound healing potential of H. italicum essential oil (HIEO) prepared in the form of ointment and gel in streptozotocin-induced diabetic wound models in rats. After creating full-thickness cutaneous wounds, forty-eight diabetic rats were divided into six groups: (1) negative control, (2) positive control, (3) ointment base, (4) gel base, (5) 0.5% HIEO ointment (6) 0.5% HIEO gel. Wound healing potential was determined by the percentage of wound contraction, hydroxyproline content, redox status, and histological observation. A significant decrease in the wound size was observed in animals treated with HIEO formulations compared with other groups. The HIEO groups also showed a higher level of total hydroxyproline content, and more pronounced restitution of adnexal structures with only the underlying muscle defect indicating the incision site. Hence, our results legitimate the traditional data of the pro-healing effect of HIEO because HIEO in both formulations such as gel and ointment exhibited the significant wound repairing effect in the incision wound model.

Multiple positive effects that forests have on human health and overall well-being have been reported widely in the literature. Still, multiple elements of this relationship remain unidentified and unexplained. In this study, the composition of leaf volatile organic compounds (BVOCs) content in three common coniferous species: the Austrian pine (Pinus nigra), Scots pine (Pinus sylvestris) and Spruce (Picea abies), was analyzed. The specificity of BVOCs content in the examined species and their genotypes is observed as a plant potential to evaporate these organic compounds and potentially improve human health and well-being. Principal component analysis applied on BVOCs content among species showed significant differences between compounds that have previously been characterized as having positive effects on human health by acting as anticancer, anti-inflammatory, antiviral and antibacterial. Variations among genotypes of the investigated species were observed in the content of BVOCs relevant for human health improvement, such as limonene, terpinolene, β-pinene, linalool, camphene, camphor, citronellol and α-cadinol. The observed intra- and inter-species variations in the BVOCs content provide an appropriate base for further research on the forest–human health relationship, breeding and selection of the most suitable genotypes for human health improvement, and could I mpact the sustainable management of forests.

The phytochemical analysis of the investigated Immortelle essential oil revealed the presence of monoterpenes and sesquiterpenes as major components that might be efficient as a wound healing potential agent. The present study aimed to develop an ointment based on the Immortelle essential oil and investigate its wound healing effects on excision wounds in diabetic rats. The topical formulated Immortelle ointment was subjected to pharmaco-technical characterization. Thirty-two diabetic rats with the induced excision wound were used to evaluate in vivo wound healing effects of ointment. The animals were randomly divided into four groups untreated or topically treated with either a 1% silver sulfadiazine, the ointment base, or Immortelle ointment. The response to the treatment was assessed by macroscopic, biochemical and histopathological analysis. The ointment, compatible with the skin remained stable for 6 months. Topical application of the Immortelle ointment showed the highest wound contraction with the highest content of hydroxyproline in comparison to the all examined groups. The Immortelle ointment showed significant wound contraction from day 7 to day 21 as compared to other groups. On the day 21, there was an average of 99.32% wound contraction in the Immortelle group, whereas the mean wound contraction in the negative control and ointment base group was 71.36% and 81.26% respectively. The histopathological results validated the potential wound healing effect of Immortelle ointment with evident post-excision scar maturation and increased collagen fibers density. Our findings revealed that the Immortelle ointment approach might serve as a promising and innovative tool for wound healing.

Medicinal plants have many beneficial effects on human health. Garlic (Allium sativum, Alliaceae) is one of the most famous herbal species, used for various diseases and conditions. Unfortunately, garlic is also associated with adverse effects, including cutaneous manifestations. In this review, burn injuries caused by application of raw garlic are reported. Searching through PubMed, Google Scholar and ResearchGate, a total of 32 articles with 39 patients were found. Demographics of patients, reasons for garlic use, details on garlic application, as well as description of burns and its treatment are thoroughly described and discussed. In most of the cases, garlic caused second-degree burns, although some circumstances can cause formation of necrotic tissue. Various body parts were affected, legs being most common. The chemistry of garlic is also presented, with focus on volatile organic sulfur compounds, which also seem to be responsible for burns formation. Treatment of garlic burns was mainly symptomatic, and various types of drugs were used. Although not commonly expected, garlic should be taken into consideration as causative agents of burns by treating doctors, and patients should be advised against application of fresh garlic onto skin and mucosa.

Introduction Human in vivo models of skin damage were often used in research of cutaneous disorders. The most commonly used models were tape-stripping as mechanical, sodium lauryl sulphate-induced irritation as chemical and ultraviolet radiation as physical damage model. In regard to differences between models, they were expected to have different responses to damage and recovery, with unique skin parameters' changes over time. Objective The aim was to compare skin parameters in three different skin damage models on the same anatomical location, with and without topical treatment. Methods Four test sites on each forearm were randomly assigned to three skin damage models with the fourth sites on each forearm chosen as a control, undamaged site. Skin parameters were assessed using non-invasive methods. Results Sodium lauryl sulphate irritation caused the strongest damage with delayed reaction to the irritant. Tape stripping leads to highest initial skin barrier disruption but afterwards it showed the fastest skin recovery. Ultraviolet radiation did not affect skin barrier function, but it elevated skin erythema and melanin level. Tested preparation did not lead to changes in measured parameters. Conclusion The skin of the participants had different response to three skin damage models with distinct changes of skin parameters and recovery. The trial was registered at ClinicalTrials.gov under the identifier NCT03783819.

St. John's wort has been used for centuries in traditional medicine of many cultures, and nowadays it is well-known as a clinically important antidepressant drug. Considering the rising market demand for Hyperici herba, quality control of crude drug is of paramount importance. In this paper we performed HPLC-DAD chemical profiling of St. John's wort tea samples obtained at local markets, pharmacies and health food stores in the Balkan Peninsula countries, Austria and Turkey. Furthermore, water alcoholic extracts of the collected samples were evaluated in terms of their antioxidant potential, as well as the ability to inhibit biologically important enzymes such as acetylcholinesterase, monoamine oxidases A and B (MAO-A and MAO-B), α-amylase and α-glucosidase. Significant variability within the samples in the quantities of hypericin, hyperforin, rutin, quercetin, gallic, chlorogenic, caffeic and p-hydroxybenzoic acid was noticed. Chemotaxonomic modelling enabled the identification of three clusters of samples based on the quantities of rutin, hypericin and hyperforin. Generally, the extracts exhibited a significant potential to inhibit MAO-A (median IC

Mints are the most popular economic and traditional herbs. The aim of this article was chemical characterization of volatile compounds from wild populations of Mentha aquatica, M. arvensis, M. longifolia, M. microphylla, M. pulegium, M. spicata, M.×dumetorum, M.×gentillis and M.×verticillata, as well as cultivated samples of M. spicata, M.×piperita 'Alba' and M.×piperita 'Crispa'. Analyses were performed directly from dried aerial parts (herb) of collected samples by headspace gas chromatography-mass spectrometry. In total 54 compounds were detected, representing from 89.99 % to 99.66 % of volatile fractions of all investigated samples. The recorded volatiles were primarily monoterpene hydrocarbons and oxygenated monoterpenes, while oxygenated aromatic monoterpenes, sesquiterpene hydrocarbons and aliphatic compounds were present in lower concentrations in analyzed samples. The major components were linalool, limonene, 1,8-cineol, α-terpinyl acetate, pulegone, β-pinene and menthol. The cluster analysis revealed five main groups or chemotypes according to qualitative and quantitative content of volatiles, as well as similarities among samples. These results contribute to the knowledge on the mints chemistry in Pannonian Plain and Balkan Peninsula.

Objective Besides conventional drugs, various vaginal disorders are often treated with complementary and alternative medicines. Different natural products are often used in treatment of genital infections. The aim is to present a case of treatment of vaginal infection with a garlic clove. Methods A detailed interview was conducted with the subject whose condition was described in this case. Results Young woman was diagnosed with vaginal Ureaplasma sp. infection. After failed local treatment with antibiotics and antifungals, subject obtained advice to treat the disorder with incised clove of garlic. Control examination by her gynecologist confirmed that infection was successfully cured. Subject experienced no adverse effects. Conclusions Garlic clove successfully cured vaginal infection presented in this case. However, application to genital organs should be reserved for dosage forms, in order to prevent potential toxicity and to obtain reproducibility of active compounds concentrations.

Graphical abstract, Constant search for new drugs with antiviral properties often extends to products of natural origin. Lamiaceae is one of the most important herbal families, well known for various biological and medicinal effects of a variety of aromatic spices, including thyme, mint, oregano, basil, sage, savory, rosemary, self-heal, hyssop, lemon balm and many others. The paper provides a review of antiviral potential of previously mentioned plants which has been demonstrated so far, with special emphasis on anti-HIV properties. Relevant articles were compiled by searching plant names combined with keywords describing antiviral activity. The antiviral effect is direct, with prominent activity against enveloped viral species. Initial stages of the viral life cycle are the most affected, as these plants appear to be targeting mainly viral structures responsible for attachment to target cells. In case of HIV, there is some activity against key enzymes in the viral life cycle. Even in the case of drug resistance, there is an equal susceptibility to applied herbal preparations. Some in vivo experiments suggest that use of Lamiaceae representatives could help in prevention and treatment of some viral diseases. A possible reduction of side effects of diseases and conventional drug therapy are also some aspects worth further investigations.

The latest report on tetrahydrocannabinol (THC) exposure assessment performed by European Food Safety Authority showed that THC levels in certain food categories are high. However, the performed study included uncertainties regarding occurrence data and hemp-food consumption patterns. The aim of the present study was to quantify the levels of “total” Δ9-THC, cannabidiol (CBD) and cannabinol (CBN) in hemp food products available on the market of countries mostly not assessed by the latest EFSA report, as well as to perform cannabinoids health risk assessment. The obtained results allowed us to identify hemp-seed oils as product type which might represent health concern. Furthermore, the analysis of hemp teas showed the possibility of Cannabis drug-type occurrence, but also questioned the justification of cannabinoids maximum transfer principle application for the exposure assessment. It was concluded, that hemp-based food products should not be consumed by population younger than 18 years.

Many studies have refuted the opinion that urban floras are poor in total species number and characterized primarily by neophytes. Also, it has been proved that urban flora differs from its surroundings, and at the same time reflects its richness. Our study focused on urban flora, structure and similarities, and the possible correlation with the urban attributes. Therefore, we selected 11 cities in Southeast Europe, which differ in the development of transportation networks (roads, rails and inland waterways) and population size. Next, we compared two sets of data (structure of urban flora and urban attributes) to define the correlation patterns. Analyzed urban floras were primarily distinguished by a total number of species, number of unique species and neophytes. Belgrade is a city with the highest urbanization level, the most species and the highest number of unique species and neophytes as well. Kosovska Mitrovica, Grocka and Požarevac stand out by floristic richness. These cities have a high number of species, yet a small proportion of neophytes. Other cities are characterized by similar floristic structure, with a relatively similar proportion of neophytes and approximately similar number of unique species in relation to the total number. It can be concluded that other factors beside the chosen urban attributes have additional effects on floristic structure, except for the factor - the presence of neophytes. This is in accordance with predefining subjects, since urban attributes were chosen based on knowledge about important corridors for invasive species.

The genus Ganoderma includes about 80 species growing on cut or rotten trees. The most commonly used species is Ganoderma ludicum. Biomolecules responsible for the health benefits of Ganoderma are polysaccharides with an immunostimulative effect and triterpenes with a cytotoxic action. For more than 2000 years, it has been used traditionally in the treatment of various pathological conditions and recently, its immunoregulatory, antiviral, antibacterial, antioxidant, hepatoprotective, and anticancer potential has been confirmed. A wide range of Ganoderma extracts and preparations arrest the cell cycle in different phases and consequently inhibit the growth of various types of cancer cells. Extracts containing polysaccharides stimulate immunological reactions through the production of various cytokines and mobilization of immune system cells. In-vivo studies have confirmed the anticancer potential and the antimetastatic effects of compounds originating from Ganoderma. There is also evidence for the chemopreventive action of Ganoderma extracts in bladder, prostate, liver, and breast cancer. The results of clinical studies suggest the combined use of G. lucidum with conventional chemotherapy/radiotherapy, but the methodology and the results of these studies are being questioned. Therefore, a constant need for new clinical trials exists.

Objective Gastrointestinal (GI) disorders are estimated to be frequent among general population. Various types of traditional and complementary therapies, primarily phytotherapy, can be used for prevention and treatment of many diseases and conditions, including GI complaints. Thus, the aim of this study was to investigate the patterns of use of medicinal herbs in treatment and prevention of GI disorders, together with their efficacy and safety. Methods A prospective, repeated cross-sectional, descriptive study was conducted in the form of a specifically created questionnaire, filled in by consumers and/or patients in pharmacies on the territory of Autonomous Province of Vojvodina, Republic of Serbia. All data were statistically analyzed in Microsoft Excel 2007. Results In the total number of 1137 patients, 10.4% declared themselves as consumers of phytopreparations for GI disorders. The most common diseases were constipation (44%) and dyspepsia (23%). The most frequently used preparations contained laxatives (with both anthraquinones and dietary fibers), artichoke and silymarin. Iberogast® was also frequently used. Pharmacists were the main source of recommendation for the most adequate herbal remedies. At the same time, phytopreparations were well tolerated, with no major side effects, and were evidently or presumably effective. Conclusions Some mild and moderate GI disorders seem to be treated frequently with phytopreparatons. Various herbal remedies are well accepted by patients, and the phytopreparations seem to have favorable ratio of safety and efficacy. Further integration into conventional medicine will improve the quality of the products used and provide a rational plan of use of medicinal plants.

The application of chemometric approaches in the comparison of the activity and the chemical composition of essential oils of 10 aromatic Lamiaceae species (Melissa officinalis, Mentha piperita, Mentha aquatica, Mentha longifolia, Ocimum basilicum, Origanum vulgare, Rosmarinus officinalis, Salvia officinalis, Salvia nemorosa, and Thymus vulgaris) is reported in this paper. The standard methodology of principal component analysis needs too many principal components (PCs) to effectively describe the relationships between studied samples. PC1 explains only 47.58% of the total variance reflecting the most significant differences between essential oils of S. nemorosa and Or. vulgare. PC2 explains 19.77% of the total variance and shows the most significant differences between the essential oils of M. aquatica compared with those of S. nemorosa, S. officinalis and Or. vulgare. Efficient compression of the data was possible by hierarchical clustering analysis (HCA) complemented with a color map. HCA shows that essential oils of both investigated representatives of genus Salvia are characterized by a high content of oxygenated sesquiterpenes and remarkable antifungal activities, but low antibacterial and antioxidant activity, particularly for the S. officinalis. Unlike sage, the essential oils of oregano and thyme have high content of aromatic oxygenated monoterpenes and low antifungal but pronounced antibacterial activity. Peppermint, basil, rosemary, and lemon balm are characterized by high content of oxygenated monoterpenes and aliphatic components as well as by high antioxidant 2,2-diphenyl-1-picrylhydrazil activity of essential oils. Copyright © 2016 John Wiley & Sons, Ltd.

Aniseed (Pimpinella anisum L., Apiaceae) and its essential oil (EO) have been widely used. Because there are some data about the impact of aniseed EO on drug effects, this survey aimed to assess the potential of pharmacokinetic herb-drug interaction between aniseed EO and acetaminophen and caffeine in mice. The chemical analysis (gas chromatography-mass spectrometry) of aniseed EO has confirmed trans-anethole (87.96%) as the main component. The pharmacokinetic studies of intraperitoneally (i.p.) and orally applied acetaminophen (200 mg/kg) and caffeine (20 mg/kg) were performed in mice after 5 days of oral treatment with human equivalent dose of aniseed EO (0.3 mg/kg/day). The analysis of pharmacokinetic data showed that in the group treated by aniseed EO, the significant decrease in the peak plasma concentration of acetaminophen after oral application (p = 0.024) was revealed when compared with control group and the reduction of systemic exposure to the drug after oral application (74 ± 32% vs. 85 ± 35% in the control) was noted. The bioavailability of orally applied caffeine was also significantly decreased (p = 0.022) after the EO treatment in comparison with the control (57 ± 24% vs. 101 ± 29%). Therefore, the compromised therapeutic efficacy of acetaminophen and caffeine during the usage of aniseed EO preparations should be considered.

Thyme (Thymus vulgaris L., Lamiaceae) is, among numerous representatives of genus Thymus, the commonly used and cultivated species worldwide. This is mainly due to the presence of essential oil which has a number of notable and scientifically confirmed biological activities. However, the amount of thyme essential oil in the leaves is very low (0.8–2.6%), thus the majority of cultivated plant material remains unused after its isolation. The aim of this study was to investigate the chemical composition of waste material extracts prepared from decoction remaining after hydrodistillation of the thyme essential oil, deodorized leaves and stems, to elucidate their potential antioxidant and hepatoprotective effects and compare obtained results with officially prepared thyme leaf extract. HPLC/DAD analysis revealed rosmarinic acid (52.36–105.8 mg/g of dried extract (d.e.)) and rutin (11.01–87.11 mg/g d.e.) as dominant compounds in all investigated thyme postdistillation waste material extracts. Also, gallic, chlorogenic and ferulic acid and apigenin were present in considerable amounts. Deodorised leaf extracts were found to possess the strong antioxidant activity tested against DPPH and OH radicals and are potent inhibitors of lipid peroxidation process in vitro. Also, investigated thyme leaves and deodorized thyme leaf extracts exhibit significant hepatoprotective effect when in vivo antioxidant activity was tested, especially in terms of xanthine oxidase, superoxide dismutase, glutathione peroxidase activity, prevention of lipid peroxidation and decrease of glutathione concentration in oxidative stress. Results of this study indicated that thyme postdistillation waste material can be used as a source of rosmarinic acid, rutin and other phenolic compounds. Also, investigated extracts could be used for the preparation of herbal medicines with antioxidant and hepatoprotective effects.

Aims The aims of study were to examine the antibacterial potential of two commercial essential oils (EOs) from coriander (Coriandrum sativum L.) and thyme (Thymus vulgaris L.) against vaginal clinical strains of bacteria and yeast and their chemical composition. Methods and Results Antimicrobial activities of commercial essential oils were determined using macro-diffusion (disc, well) and micro-dilution method in 96-well micro plates against twelve clinical strains of bacteria: Escherichia coli, Proteus mirabilis, Staphylococcus aureus and Enterococcus sp., Staph. aureus ATCC 25923, ATCC 6538 and E. coli 25922 and two clinical Candida albicans strains, including ATTC 10231. Spectrophotometric method was used for determination on C. albicans growth. An antimicrobial effect of EOs was strain specific. Bactericidal activity was higher for coriander EO (minimal inhibitory concentration (MICs) 0·4–45·4 μl ml−1) against almost all tested bacteria, except multiple resistant strains of Eneterococcus sp. and Proteus sp. Thyme EO showed slightly better fungicidal activity reaching MIC at 0·11 mg ml−1 for all C. albicans strains. The effect of EOs on biofilm-forming ability was tested for two strains of Staph. aureus and E. coli, as well as on C. albicans filamentation ability. Brine shrimp lethality bioassay revealed thymus oil total toxicity and coriander oil intoxicity (LC50 = 2·25 mg ml−1). The chemical composition of oils was analysed by gas chromatography mass spectrometry showing oxygenated monoterepenes as dominant constituents. Conclusions The results provide in-vitro scientific support for the safety possible use of Coriander EO against E. coli, Staph. aureus and C. albicans vaginal infections in alternative gynaecological treatment. Significance and Impact of Study To examine EOs as possible constituent of naturally based antimicrobial agents in vaginaletes for safety gynaecological application.

Numerous traditional and clinically approved indications have created constant raise of demand for Hyperici herba in the world. However, the quality of the plant material depends on the plant source and abiotic factors of the habitats. The scope of the research was to find out whether other Hypericum species have similar chemical characteristics for potential medical use. Hypericum maculatum Crantz is in some Pharmacopoeias listed together with Hypericum perforatum L. as biological source of H. herba, because of similar chemical composition. Often the full taxonomic identification stating the used subspecies of H. maculatum is being avoided. Therefore, the aim of study was to chemically characterize water–alcoholic extracts of subspecies Hypericum maculatum Crantz subsp. maculatum. The samples were collected at three localities, with different habitat characteristics during three ontogenetic phases. The amounts of total phenolics and flavonoids were estimated, as well as the amounts of ferulic and chlorogenic acid, rutin, quercitrin, amentoflavone, apigenin, hypericin, and hyperforin. Chemical composition and ecological characteristics were analyzed by PCA methods. Source of sunlight and soil acidity were shown as ecological factors that potentially could be cause for chemical profiling of samples. Biological potential of extracts was estimated through antioxidant assays and inhibition of acetylcholinesterase (AChE) and α-amylase with the aim to explore potential use of plant in the treatments of diabetes and Alzheimer’s disease. All the extracts exhibited notable antioxidant potential in the used test systems as well as the ability to inhibit AChE and α-amylase. H. maculatum subsp. maculatum exhibits notable correspondence in chemical composition and biological activity with H. perforatum.

St. John’s wort is a widely used medicinal plant. The quality of herbal drug, which is in most of the cases collected from nature, varies. Therefore, the aim of the present study was detailed chemical characterization of Hypericum perforatum subsp. perforatum samples collected in close time intervals during flowering and fruitification with the purpose to state the phenological stage characterized by maximum levels of active principles. The antioxidant potential and potential to inhibit biologically important enzymes, as well as the cytotoxicity and genotoxicity of the sample collected during the full flowering period, were evaluated. Data showed that the optimal period for the achieving of maximum level of active principles is the phenophase between floral budding and flowering stage. Significant antioxidant potential and the ability to inhibit biologically important enzymes (especially α-glucosidase) were recorded. The extract exhibited no genotoxicity in subcytotoxic concentrations, while increased cytotoxicity recorded in cotreatment with bleomycin on malignant cell lines was especially significant.

Oxidative stress has been associated with cervical cancer. Our aim was to examine lipid peroxidation and the extent of oxidative stress in women diagnosed with different stages of cervical cancer in order to evaluate its potential role in the evolution of cancer.We measured the concentration of thiobarbituric acid reactive substances, activities of antioxidative enzymes and 8-hydroxy-2-deoxyguanosine in 153 subjects. Enzymatic activity as well as TBARS concentration were measured spectrophotometrically, while 8-OHdG was determined by gas chromatography-mass spectrometry. PPatients were categorized: group II H-SIL; group III FIGO Ia-Ib and group IV FIGO IIa-IV.Our results showed highly significant increase in the level of lipid peroxidation in group IV when com pared to the control group, group II and group III (p0.001). Activity of superoxide dismutase was also significantly higher in group IV when compared to control group (p0.01), group II (p0.01) and group III (p0.05). Activity of catalase was also significantly higher in group IV when compared to control group (p0.005), group II (p0.005) and group III (p0.05). Activity of glutathione-S-transferase was also significantly higher in group IV when compared to control group (p0.05), group II (p0.05) and group III (p0.05). Activities of glutathione peroxidase and glutathione reductase showed no significant differences among the groups. Level of 8-OHdG was significantly higher in group IV than in the other groups (p0.01).It can be concluded that oxidative stress is possibly involved in the pathogenesis of cervical cancer, demonstrated by increased lipid peroxidation and an altered antioxidant defense system and higher levels of 8-OHdG.Utvrđeno je da postoji veza između oksidativnog stresa i cervikalnog karcinoma. Naš cilj bio je da ispitamo lipidnu peroksidaciju i nivo oksidativnog stresa kod žena kojima je dijagnostikovan cervikalni karcinom u različitim fazama da bi se otkrila njegova eventualna uloga u razvoju karcinoma.Merili smo koncentraciju reaktivnih supstanci tiobarbiturne kiseline, aktivnosti antioksidativnih enzima i 8-hidroksi-2-deoksigvanozina kod 153 ispitanice. Enzimska aktivnost kao i koncentracija TBARS merene su spektrofotometrijski, dok je 8-OhdG određen pomoću gasne hromatografije – masene spektrometrije. Pacijentkinje su podeljene u kategorije: grupu II H-SIL; grupu III FIGO Ia-Ib i grupu IV FIGO IIa-IV.Naši rezultati pokazali su vrlo značajan porast nivoa lipidne peroksidacije u grupi IV u poređenju s kontrolnom grupom, grupom II i grupom III (p0,001). Aktivnost superoksid-dismutaze bila je takođe značajno viša u grupi IV u poređenju s kontrolnom grupom (p0,01), grupom II (p0,01) i grupom III (p0,01). Aktivnost katalaze bila je isto značajno viša u grupi IV u poređenju s kontrolnom grupom (p0,005), grupom II (p0,005) i grupom III (p0,05). Aktivnost glutation-S-transferaze je takođe bila značajno viša u grupi IV u poređenju s kontrolnom grupom (p0,05), grupom II (p0,05) i grupom III (p0,05). Aktivnosti glutation peroksidaze i glutation reduktaze nisu ukazale na značajne razlike između grupa. Nivo 8-OHdG bio je značajno viši u grupi IV nego u ostalim grupama (p0,01).Može se zaključiti da oksidativni stres potencijalno učestvuje u patogenezi cervikalnog karcinoma, na šta ukazuju povišena lipidna peroksidacija i promene su sistemu antioksidativne zaštite i povišen nivo 8-OHdG.

The metabolic syndrome (MetS) is a polygenic multifactorial metabolic disorder with strong socioeconomic influence. MetS has became a worldwide epidemic, that directly increases the risk of cardiovascular diseases and type 2 diabetes mellitus. The human apoE gene, coding Apolipoprotein E, has three common polymorphisms in human population: e2, e3 and e4, which are proved to be associated with impaired lipid metabolism. The contribution of apoE polymorphism to MetS disorders has not been investigated previously in Vojvodina Province, region with the highest number of obese people in Serbia. The aim of this study was to evaluate apoE gene polymorphism in relation to MetS disorders. The healthy control group of 30 individuals and 63 MetS patients were examined for apoE variants in relation to biochemical and anthropometric parameters. The genotypes were determined by PCR-RFLP. Regarding all parameters, significantly higher values were detected in MetS group compared to control. The MetS group of patients had significantly higher frequency of e4 allele. In addition, positive relation was revealed between e4 allele presence and all measured parameters. It was found that the e4 allele was related with a significantly increased OR of MetS disorders according to the International Diabetes Federation definition. These results suggested that e4 allele may act as a one of determinants for development of metabolic syndrome.

The present study was designed to define the phenolic and terpene profile, antioxidant and anti-inflammatory activity of methanol extracts and essential oils of the leaves and seed cones of Juniperus foetidissima species, on which there has been little previous research. The content of 44 phenolics in extracts was determined using LC–MS/MS. A comparable qualitative composition, including dominant compounds catechin, quercitrin, epicatechin, rutin and quercetin-3- O -glucoside, was found in both extracts. GC–MS analysis showed a simple terpene composition, with monoterpenes dominant and sabinene most abundant compound, 39.9% in leaves and 29.9% in seed cones. Antioxidant activity was determined by several assays. All results of antioxidant tests were comparable to BHT, a well-known synthetic antioxidant. A novel method for estimation of anti-inflammatory potential by means of inhibition of COX-1 and 12-LOX activity was described. Essential oils showed significant anti-inflammatory activity, especially of leaves. To conclude, J. foetidissima showed great bio potential, and its usage in production of food and beverages should be considered.

The study shows the influence of the origin of plant material and biological source on the in vitro antioxidant (neutralization of DPPH and OH radical, nitric oxide, and inhibition of lipid peroxidation) and anticholinesterase activity of chemically characterized and quantified ethanol extracts of ten St. John’s wort samples. The investigated samples were: five Hypericum perforatum species representatives collected at different localities, one commercial sample of Hyperici herba purchased at a local market and four Hypericum species autochtonous to the Balkan Peninsula (H. maculatum subsp. immaculatum, H. olympicum, H. richeri subsp. grisebachii and H. barbatum). All the examined extracts exhibited notable antioxidant potential, but in most of the cases indigenous Hypericum species expressed stronger effects compared to the original source of the drug, H. perforatum. The changes in the content of phenolic compounds, especially flavonoids, hyperforin and hypericin, related to the source of the drug affected the investigated activities. Since all of the investigated species have shown prominent inhibition of acetylcholinesterase in vitro activity, they could be further investigated as potential substances in preventing of Alzheimer’s disease.

Anise (Pimpinella anisum L.; Apiaceae) and its essential oil have been widely used in folk medicine, pharmacy and food industry. Since there are some data about the impact of anise on functions of central nervous system (CNS), the issue of possible interactions with drugs acting in CNS should be considered. This survey aimed to examine the influence of aniseed essential oil (EO) intake on the effects of drugs that act in CNS. The chemical profile of essential oil determined by GC-MS revealed as the main components: trans-anethole (88.49%), γ-himachalene (3.13%), cis-isoeugenol (1.99%), and linalool (1.79%). The effects of codeine, diazepam, midazolam, pentobarbital, imipramine and fluoxetine were tested in mice after 5days of peroral pretreatment with human equivalent dose of aniseed EO (0.3mg/kg). The intake of EO led to significant increase of analgesic effect of codeine. The motor impairment caused by midazolam was enhanced in the group treated by EO. The application of diazepam decreased the number and percentage of entries in open arm in elevated maze plus test in the group pretreated with EO indicating augmented effect of drug on motor activity. EO pretreatment caused significant shortage of pentobarbital induced sleeping time when compared to control. The decrease in antidepressant effect of imipramine and fluoxetine was diminished by the pretreatment with aniseed EO. Based on the results of this study we conclude that concomitant intake of aniseed EO preparations and drugs that act on CNS should be avoided due to potential herb-drug interactions, which also need further clinical confirmation.

The present study was designed to define the phenolic profile, antioxidant, anti-inflammatory and cytotoxic activity of Plantago altissima L., which has never been studied before and to compare it with closely-related, renowned, well-studied Plantago lanceolata L. The presence and content of 44 phenolics in methanol extracts were studied using LC–MS/MS. A similar qualitative composition including dominant compounds as p-hydroxybenzoic, vanillic, gallic and chlorogenic acid, besides apigenin, luteolin and luteolin-7-O-glucoside was found between both extracts. Antioxidant activity of extracts was determined using several assays. All results of these tests were comparable to butylated hydroxytoluene, a well-known synthetic antioxidant. Anti-inflammatory potential was studied by means of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibitory activity. Activity of P. altissima towards COX-1/12-LOX inhibition (IC50 = 4.4 and 3.6 mg/mL, respectively) was inferior to activity of P. lanceolata (IC50 = 2.0 and 0.8 mg/mL, respectively). Treatment of four cell lines resulted in a considerable dose-dependent inhibition of cell growth, where P. lanceolata exerted a stronger effect (IC50 = 172.3, 142.8, 405.5 and 551.7 μg/mL for HeLa, MCF7, HT-29 and MRC-5 cell lines, respectively). To conclude, P. altissima showed certain bio-potential, but was clearly inferior to P. lanceolata.

The appearance of common and self-initiative usage of various herbal preparations in everyday practice and life imposes the question of possible interactions with drugs. This survey examined the influence of acute and chronic peppermint oil (PO – Mentha × piperita L., Lamiaceae; prepared as emulsion for oral use) on pentobarbitone-induced sleeping time, analgesic effect of codeine and impairment of motor coordination caused by midazolam in mice. The chemical profile of essential oil was determined by GC-MS. Applied doses of PO were 0.1 and 0.2 mL/kg. Chronic PO intake (in both doses) led to significant decrease of analgesic effect of codeine, while acute intake of PO did not change this effect. Acute PO pretreatment in higher dose caused significant prolongation of pentobarbitone-induced sleeping time, while it was significantly shortened by chronic PO pretreatment at the same dose. Midazolam effect was enhanced and prolonged significantly by chronic PO intake at higher dose, while acute intake of PO did not change this effect. Gut motility was increased only by acute intake of higher PO dose. Regarding the fact that PO produces changes in tested drug effects, the interaction between drugs and phytopreparations containing PO should be additionally followed/confirmed in humans. Copyright © 2011 John Wiley & Sons, Ltd.

The antioxidant properties of methanol extracts of Lady’s Bedstraw (Galium verum L., Rubiaceae) herb from two different localities in Serbia were evaluated. Antioxidant activity was assessed in four different model systems. Free radical scavenging capacity (RSC) was examined by measuring the scavenging activity of extracts on 2,2-diphenyl-1-pycrylhydrazil (DPPH) and hydroxyl radical (OH), as well as on hydrogen peroxide. In addition, the protective effects of lipid peroxidation (LP) in corn oil were evaluated by the TBA-assay using the Fe2+/ascorbate system of induction. The amount of dried extract, the content of total phenolics, flavonoids and chlorophylls was also determined. Extracts from both locations expressed very strong scavenger activity, reducing the DPPH⊙ (IC50=3.10 µg/mland 8.04 µg/ml) and OH radical formation (IC50=0.05 µg/ml and 0.54 µg/ml) and neutralising H2O2 (IC50=4.98 µg/ml and 3.80 µg/ml), in a dose dependant manner. Also, examined extracts showed notable inhibition of LP (IC50=11.69 µg/ml and 19.47 µg/ml). The observed differences in antioxidant activity could be partially explained by the levels of phenolics (2.44–4.65 mg and 4.57–5.16 mg gallic acid equivalents/g dry extract), flavonoids (6.38–10.70 µg and 15.56–17.96 µg quercetin equivalents/g dry extract) and chlorophylls in the investigated Lady’s Bedstraw extracts.

Aniseed (Pimpinella anisum L., Apiaceae) and its essential oil (EO) have been widely used. Because there are some data about the impact of aniseed EO on drug effects, this survey aimed to assess the potential of pharmacokinetic herb-drug interaction between aniseed EO and acetaminophen and caffeine in mice. The chemical analysis (gas chromatography-mass spectrometry) of aniseed EO has confirmed trans-anethole (87.96%) as the main component. The pharmacokinetic studies of intraperitoneally (i.p.) and orally applied acetaminophen (200 mg/kg) and caffeine (20 mg/kg) were performed in mice after 5 days of oral treatment with human equivalent dose of aniseed EO (0.3 mg/kg/day). The analysis of pharmacokinetic data showed that in the group treated by aniseed EO, the significant decrease in the peak plasma concentration of acetaminophen after oral application (p = 0.024) was revealed when compared with control group and the reduction of systemic exposure to the drug after oral application (74 ± 32% vs. 85 ± 35% in the control) was noted. The bioavailability of orally applied caffeine was also significantly decreased (p = 0.022) after the EO treatment in comparison with the control (57 ± 24% vs. 101 ± 29%). Therefore, the compromised therapeutic efficacy of acetaminophen and caffeine during the usage of aniseed EO preparations should be considered.

The chemical composition and antioxidant properties of the essential oil and EtOH extract of immortelle (Helichrysum italicum (Roth) G.Don subsp. italicum, Asteraceae) collected in Montenegro were evaluated. The essential oil was characterized by GC/MS analysis, and the content of total phenolics and flavonoids in the EtOH extract was determined using the FolinCiocalteu reagent. The free-radical-scavenging capacity (RSC) of both the essential oil and the EtOH extract was assessed with the 2,2-diphenyl-1-pycrylhydrazyl (DPPH) method. Moreover, the inhibition of hydroxyl radical ((.) OH) generation by the EtOH extract of immortelle was evaluated for the first time here. Neryl acetate (28.2%) and γ-curcumene (18.8%) were the main compounds in the essential oil, followed by neryl propionate (9.1%) and ar-curcumene (8.3%). The chemical composition of the oils of the examined and additional 16 selected Helichrysum italicum taxa described in literature were compared using principal component (PCA) and cluster (CA) analyses. The results of the statistical analyses implied the occurrence of at least four different main and three subchemotypes of essential oils. Considering the antioxidant properties, the EtOH extract of immortelle exhibited similar potential as propyl gallate and quercetin, while the essential oil exhibited relatively weak DPPH(.) -scavenging capacity.

Eterična olja origana (Origanum vulgare L.), mete (Mentha piperita L.), janeža (Foeniculum vulgare Mill.), in borovih (Abies alba Mill.) iglic in storžev, ter njihove sestavine timol, karvakrol, mentol in anisaldehid smo testirali kot snovi z protiglivno aktivnostjo proti plesnim vrste Penicillium verrucosum. Najnižje minimalne inhibitorne koncentracije (MICs) smo določili eteričnemu olju origana, ter karvakrolu, timolu, in mentolu. Rast plesni vrste P. verrucosum je bila popolnoma inhibirana po 21 dneh inkubacije v prisotnosti ½ MIC timola (0,0625 mg mL-1), medtem ko so eterično olje origana (½ MIC, ¼ MIC), timol (¼ MIC) in karvakrol (½ MIC, ¼ MIC) inhibirali rast plesni od 8,3 % do 70.8 %. Dodatek mentola v koncentracijah ¼ MIC in ½ MIC (0.1875 mg mL-1 in 0.3750 mg mL-1) pa ni imel inhibitornega vpliva na rast plesni vrste P. verrucosum. Popolno inhibicijo tvorbe ohratoksina A smo določili le v primeru, ko je bila tudi rast teh plesni popolnoma inhibirana (pri ½ MIC timola). Največjo inhibicijo tvorbe ohratoksina A (96,9 %) po 21 dnevih inkubacije smo določili pri ¼ MIC timola (0.0313 mg mL-1). Pospešeno tvorbo ohratoksina A (od 13,9 % do 28,8 %) pa smo določili pri ¼ MIC eteričnega olja origana (0.2930 mg mL-1) in ½ MIC karvakrola (0.1953 mg mL-1). Protiglivni vplivi so bili odvisni od testirane snovi, njene koncentracije ter časa interakcij med protiglivno snovjo in plesnimi vrste P. verrucosum. Rezultati nakazujejo možnosti uporabe eteričnega olja origana v živilih namesto kemijskih konzervansov. Ker pa so živila kompleksna okolja v katerih je potrebno upoštevati več dejavnikov, so potrebne nadaljnje raziskave., Essential oils from oregano (Origanum vulgare L.), mint (Mentha piperita L.), fennel (Foeniculum vulgare Mill.), and pine (Abies alba Mill.) needles and cones, and their active substances thymol, carvacrol, menthol, and anisaldehyde were tested for antifungal activity against Penicillium verrucosum. The lowest minimal inhibitory concentrations (MICs) were achieved for essential oil of oregano, followed by carvacrol, thymol, and menthol. These antifungal components were further investigated, as the main aim of our study was to elucidate the effect of natural antifungals on ochratoxin A production. During 21 days of exposure, the growth of P. verrucosum, and subsequently the production of ochratoxin A, was fully inhibited by thymol at ½ MIC (0.0625 mg mL-1), but menthol at ¼ and ½ MIC (0.1875 and 3750 mg mL-1) showed no growth inhibition. After 21 days of incubation, the greatest inhibitory effect on ochratoxin production (inhibition was 96.9 %) was also achieved with thymol at ¼ MIC (0.0313 mg mL-1). Essential oil of oregano (¼ MIC, 0.2930 μL mL-1) and carvacrol (½ MIC, 0.1953 μL mL-1) stimulate production of ochratoxin A at 13.9 % to 28.8 %, respectively. The observed antifungal effects depended on the agent, the concentration used, and the time of interaction between the agent and P. verrucosum. Our results indicate the possibility of using oregano essential oil as a substitute for artificial preservatives in certain foods, but further research is needed.