Evaluation of the Anti-Inflammatory/Immunomodulatory Effect of Teucrium montanum L. Extract in Collagen-Induced Arthritis in Rats.

The anti-inflammatory/immunomodulatory effects of Teucrium montanum L. (TM), a plant distributed in the Mediterranean region, have been insufficiently examined. The effects of the TM ethanol extract were tested in a rat collagen-induced arthritis (CIA) model of rheumatoid arthritis. LC-MS was used for the phytochemical analysis of the TM extract. Dark Agouti rats were immunized with bovine type II collagen (CII) in incomplete Freund's adjuvant for CIA, and treated with 100 or 200 mg/kg of TM extract daily via oral administration. Clinical and histopathological evaluations and a flow cytometric analysis of the phenotypic and functional characteristics of splenocytes and draining lymph node cells were performed. The cytokines in the paw tissue culture supernatants and anti-CII antibodies in serum were determined by ELISA. The TM extract, with the dominant components verbascoside and luteolin 7-O-rutinoside, reduced the arthritic score and ankle joint inflammation in CIA rats, promoted the antioxidant profile in serum, and lowered pro-inflammatory TNF-α, IL-6 and IL-1ß production. It suppressed the activation status of CD11b+ cells by lowering CD86, MHCII and TLR-4 expression, and promoted the Th17/T regulatory cell (Tregs) balance towards Tregs. A lower frequency of B cells was accompanied by a lower level of anti-CII antibodies in treated rats. These findings imply the favorable effect of TM extract on the clinical presentation of CIA, suggesting its anti-inflammatory/immunomodulatory action and potential therapeutic effect.

Nutritional Value, Phytochemical Composition and Biological Activities of Lycium barbarum L. fruits from Serbia.

Cultivation of goji berries (GB), fruits of Lycium barbarum L. (Solanaceae), is expanding worldwide, including in Europe. In this study, a comparative analysis of the nutritional value, chemical composition and in vitro biological activities of GB from different locations in Serbia was performed. Proximate compositions were evaluated according to standard methods. Minerals were assessed by inductively coupled plasma techniques, while fatty acids, sterols, and phenolic profiles were analyzed by gas- and liquid chromatography-based techniques coupled with flame-ionization, mass spectrometry, or diode array detection. The total content of phenolics, flavonoids, carotenoids, and polysaccharides was assessed using spectrophotometric methods. Methanol extracts from GB were examined for their antioxidant, enzyme inhibitory (α-amylase, α-glucosidase, acetylcholinesterase and tyrosinase) and antibacterial activities. Despite significant variations among samples from different locations, the results confirmed that GB are a valuable source of dietary fiber and protein and are characterized by favorable fatty acid profiles. Phytochemical analysis revealed that ß-sitosterol, Δ5-avenasterol, and 24-methyldesmosterol are the predominant sterols and caffeic acid, gallic acid, quercetin and rutin are the main phenols. All GB samples showed both antioxidant and mild antimicrobial activity. A dose-dependent anti-enzymatic activity (IC50 ranging 1.68-6.88 mg/mL) was demonstrated. The results support further promotion of GB cultivation in Serbia and further investigations on their potential applications in various industries.

Health Benefits and Applications of Goji Berries in Functional Food Products Development: A Review.

Goji berries have long been used for their nutritional value and medicinal purposes in Asian countries. In the last two decades, goji berries have become popular around the world and are consumed as a functional food due to wide-range bioactive compounds with health-promoting properties. In addition, they are gaining increased research attention as a source of functional ingredients with potential industrial applications. This review focuses on the antioxidant properties of goji berries, scientific evidence on their health effects based on human interventional studies, safety concerns, goji berry processing technologies, and applications of goji berry-based ingredients in developing functional food products.

Phenolic Composition, Antioxidant, Anti-Enzymatic, Antimicrobial and Prebiotic Properties of Prunus spinosa L. Fruits.

Blackthorn (Prunus spinosa L.) fruit is bluish-black wild fruit traditionally used in nutrition and medicine. It is recently gaining attention as a functional food and an underutilized source of bioactive compounds for application in the food and pharmaceutical industry. This study aimed to assess the health-promoting potential of blackthorn fruits from Serbia by examining their chemical composition and in vitro biological activities. Phytochemical analysis of the blackthorn fruit extracts was performed using LC-DAD-ESI-MS. The total phenolic (TPC), total flavonoid (TFC), total anthocyanin (TAC) content, antioxidant capacity, and enzyme inhibitory activities were determined spectrophotometrically. The antimicrobial and prebiotic properties were tested using the broth microdilution method. Twenty-seven phenolics belonging to the classes of hydroxybenzoic and hydroxycinnamic acids derivatives, flavonoids, and anthocyanins were identified, with caffeoylquinic acid as the most abundant compound. Blackthorn extracts were characterized by notable TPCs, TFCs, and TACs, and free radical scavenging and reducing ability. The enzyme inhibitory effects (IC50 = 0.43-2.16 mg/mL) were observed towards α-amylase, α-glucosidase, acetylcholinesterase, and tyrosinase. Blackthorn fruit extracts in a concentration-dependent manner (0.3-5 mg/mL) stimulated the growth of several probiotic microorganisms and their mixtures, especially the yeast Saccharomyces boulardii. Obtained results support further evaluation of the functional food potential of blackthorn fruit.

Chemical Characterization, Antioxidant and Antimicrobial Properties of Goji Berries Cultivated in Serbia.

Since the fruits of Lycium L. species (Fructus Lycii, goji berries) are promoted as a "superfood" with plenty of health benefits, there is extensive research interest in their nutritional and phytochemical composition. In the present study, the nutritional value, minerals, fatty acid composition, and bioactive compounds of L. barbarum L., red, yellow, and black goji berry (L. ruthenicum Murray.) cultivated in Serbia were investigated. Antioxidant and antimicrobial properties of their methanol extracts were assessed. Red goji berry had the highest content of fats, dietary fiber, iron, total carotenoids, and 2-O-ß-d-glucopyranosyl-l-ascorbic acid (AA-2ßG). The yellow goji berry extract showed the highest level of flavonoids and the most prominent antimicrobial (especially against Gram-negative bacteria) properties. The highest total phenolic content and the most potent antioxidant activity were observed for the extract of black goji berry. Therefore, all goji berries could be a valuable source of bioactive compounds in the food and pharmaceutical industry.

Chemical Composition of Volatiles of Eight Geranium L. Species from Vlasina Plateau (South Eastern Serbia).

Geranium species are widely used in traditional medicine of Balkan. The aim of this work was to investigate and compare chemical composition of volatile fractions obtained by hydrodistillation from aerial parts of G. macrorrhizum, G. phaeum, G. sanguineum, G. robertianum, G. palustre, G. pyrenaicum, G. columbinum and G. lucidum as well as from underground parts of G. macrorrhizum and G. phaeum, originated from Vlasina plateau in South Eastern Serbia. The volatiles were analyzed using GC/MS and GC-FID. G. palustre volatiles have been studied for the first time with ß-selinene (18.6 %) as a characteristic compound. The cluster analysis revealed separation of volatiles into two main groups. Volatile fractions of G. macrorrhizum were separated from all other samples due to high sesquiterpene content (92.3 % in aerial and 94.6 % in underground parts). The volatile fractions of other samples were mainly composed of sesquiterpenes (10.8-61.8 %), diterpenes (12.9-43.0 %) and fatty acids and their derivatives (6.6-21.6 %) with the exception of volatile fraction of G. phaeum underground parts which was dominated only by fatty acids and their derivatives (76.6 %). The results presented in this article contribute to the knowledge on the chemistry of this genus and advances the knowledge on flora of southeast Serbia.

Influence of different wild-garlic (Allium ursinum) extracts on the gastrointestinal system: spasmolytic, antimicrobial and antioxidant properties.

OBJECTIVES: As there are no previous studies of the European wild-garlic (Allium ursinum) effects on the gastrointestinal system, despite its traditional applications in gastrointestinal disorders' treatment and regular use in the human diet, we have quantified and compared spasmolytic, antimicrobial and antioxidant activities of its different leaf extracts. METHODS: Wild-garlic extracts were tested for spasmolytic activity on isolated rat ileum, antimicrobial activity on selected Gram-positive and Gram-negative bacteria and fungi by microdilution method and antioxidant capacity by DPPH radical-scavenging assay. KEY FINDINGS: Wild-garlic extracts were found to decrease ileal basal tone. As the relaxation of K+ -induced contractions was similar to one caused by papaverin, the observed spasmolytic effect was most likely mediated through Ca2+ -channel inhibition. Ethanolic extract (with the highest phenolic and high alk(en)yl cysteine sulphoxides' levels) produced the strongest spasmolytic activity. In case of acetylcholine-induced contractions, only hydromethanolic extract showed no statistical difference in comparison with positive control. All samples exhibited certain antioxidant potential and strong antimicrobial activity against tested enteropathogenic strains (Salmonella enteritidis was the most sensitive, followed by Escherichia coli, Proteus mirabilis and Enterococcus faecalis). CONCLUSION: Besides other already established health-promoting effects, wild garlic could be useful in treatment of mild gastrointestinal disturbances.

Habitat-related variation in composition of the essential oil of Seseli rigidum Waldst. & Kit. (Apiaceae).

Plant specialised metabolites like essential oils are highly variable depending on genetic and various ecological factors. The aim of the present work was to characterise essential oils of the species Seseli rigidum Waldst. & Kit. (Apiaceae) in various organs on the individual and populational levels. Geographical variability and the impact of climate and soil type on essential oil composition were also investigated. Individually sampled essential oils of roots, aerial parts and fruits of plants from seven populations were analysed by GC-FID and GC-MS. The investigated populations showed high interpopulational and especially intrapopulational variability of essential oil composition. In regard to the variability of essential oils, different chemotypes were defined. The essential oils of S. rigidum roots represented a falcarinol chemotype, oils of aerial parts constituted an α-pinene or α-pinene/sabinene chemotype and fruit essential oils can be characterised as belonging to a complex sabinene/α-pinene/ß-phellandrene/falcarinol/germacrene B chemotype. At the species level, analysis of variance (ANOVA), principal component analysis (PCA) and canonical discriminant analysis (CDA) showed that the plant part exerted the strongest influence on the composition of essential oils. Climate had a high impact on composition of the essential oils of roots, aerial parts and fruits, while influence of the substrate was less pronounced. The variations in main compounds of essential oils based on climate or substrate were complex and specific to the plant part.

Chemical Composition and Antimicrobial Activity of the Essential Oil and Methanol Extract of Hypericum aegypticum subsp. webbii (Spach) N. Robson.

Hypericum aegypticum subsp. webbii is an evergreen shrub spread in Mediterranean part of central and southeastern Europe. The chemical composition and antimicrobial activity of the essential oil and MeOH extract of H. aegypticum subsp. webbii were investigated. The monoterpenes α-pinene (63.4 - 68.5%) and ß-pinene (16.9 - 17.0%) were main compounds in the volatile oil from aerial parts. In the cluster analysis, the essential oil of H. aegypticum subsp. webbii was separated and chemically different from the oil of other subspecies of H. aegypticum as well as other Hypericum species from Greece. SIMPER analysis revealed that α-pinene (24.79%) was the component that contributed the most to differences between all oils. Also, there was extremely high overall dissimilarity between three subspecies of H. aegypticum. MeOH extract of aerial parts of H. aegypticum subsp. webbii contained flavonoids rutin (56.4 ± 0.9 mg/g), hyperoside and quercetin, and phenolic acids chlorogenic and caffeic acid, while naphthodianthrones were not detected. The antimicrobial activity of essential oil was moderate (MIC from 100 to >200 µg/ml), while MeOH extract inhibited the growth of Gram-positive bacteria Bacillus subtilis, Enterococcus faecalis, Staphylococcus epidermidis, and Micrococcus luteus (MIC 50 - 100 µg/ml), more pronounced than the extract of H. perforatum (MIC 200 - >200 µg/ml).

Seasonal variations in the composition of the essential oils of Lavandula angustifolia (Lamiacae).

Seasonal variations in the composition of the essential oils obtained from the same individual (of the same genotype) of Lavandula angustifolia cultivated in Belgrade were determined by GC and GC/MS. The main constituents were 1,8-cineole (7.1-48.4%), linalool (0.1-38.7%), bomeol (10.9-27.7%), beta-phellandrene (0.5-21.2%) and camphor (1.5-15.8%). Cluster analysis showed that the 21 samples collected each month during the vegetation cycle were separable into three main clades with different compositions of essential oils. In the shoots with flowers, inflorescences and fruits of clade I, linalool is dominant, in the young leaves before flowering and old leaves of clade II, 1,8-cineole is dominant. In the young and incompletely developed leaves of clade III, beta-phellandrene is dominant. The composition of the essential oils of lavender depended on the plant part and the stage of development.

Variability of the root essential oils of Seseli rigidum Waldst. & Kit. (Apiaceae) from different populations in Serbia.

The chemical compositions of the essential oils of seven natural populations of Seseli rigidum were analyzed. The essential-oil yield ranged from 0.16 to 2.09%. Analysis of variance (ANOVA) revealed that there were no statistically significant differences in the mean essential-oil yields between the populations, and no significant influence of the climate or soil type on the oil yield was observed. In all 67 analyzed samples, the polyacetylene falcarinol was the main compound, followed by octanal, methyl linoleate, α-muurolene, 3-butylphthalide, falcarinone, muurola-4,10(14)-dien-1ß-ol, ß-sesquiphellandrene, salvial-4(14)-en-1-one, δ-amorphene, spathulenol, and isospathulenol. The principal component analysis (PCA), the canonical discriminant analysis (CDA), and the cluster analysis (CA) revealed differentiation between the populations based on the climate. Three groups of populations were formed; the first group was composed of samples growing in regions with a humid climate, with oils having high falcarinol and low sesquiterpene contents, and the second and third groups comprised samples exposed to semi-arid climate, with oils characterized by a lower falcarinol and higher α-muurolene, δ-amorphene, ß-sesquiphellandrene, and salvial-4(14)-en-1-one contents. The semi-arid populations were divided into two groups, which were distinguished based on the oil contents of sesquiterpenes, falcarinone, and 3-butylphthalide. On the other hand, no clear separation between populations based on the different soil types could be observed.

Antimicrobial, antioxidant and anti-inflammatory activity of young shoots of the smoke tree, Cotinus coggygria Scop.

In this study, we investigated the antimicrobial activity of the young shoots of the smoke tree, Cotinus coggygria Scop., Anacardiaceae. The acetone extract and the derived ethyl acetate fraction effectively inhibited the growth of Gram-positive and Gram-negative bacteria (MIC 25-200 µg/ml), while the chloroform fraction showed pronounced activity against the yeast Candida albicans (MIC 3.12 µg/ml). The ethyl acetate fraction exhibited a significant ferric-reducing ability (10.7 mmol Fe(2+) /g extract), a very high DPPH radical scavenging activity (SC50 = 1.7 µg/ml) and inhibition of lipid peroxidation (IC50 = 41.8 µg/ml). High amounts of total phenolics (929.8 mg/g), tannins (833.8 mg/g) and flavonoids (35.5 mg/g) were determined in the ethyl acetate fraction, which also exerted significant anti-inflammatory (76.7%) and cytotoxic effects (IC50 = 15.6 µg/ml).

Chemical composition and antimicrobial activity of essential oil of different parts of Seseli rigidum.

The chemical composition and antimicrobial activity of the essential oil of the Balkan endemic species Seseli rigidum Waldst. & Kit. (Apiaceae) was investigated. The monoterpene alpha-pinene was predominant in the volatile oil from aerial parts (57.4%) and fruit (23.3%). In the essential oil of the aerial parts limonene (6.7%), camphene (5.8%) and sabinene (5.5%) were also present in high amounts, and in the fruit oil, beta-phellandrene (17.4%) and sabinene (12.9%). On the contrary, the root essential oil was composed almost entirely of the polyacetylene falcarinol (88.8%). The antimicrobial activity of the root essential oil was significant against Staphylococcus aureus, S. epidermidis, Micrococcus luteus and Enterococcus faecalis (MICs 6.25-25.00 microg/mL). Volatile constituents from the root strongly inhibited the growth of methicillin-resistant strains of S. aureus (MICs 6.25-50.00 microg/mL). Anti-staphylococcal activity can be attributed to the main volatile constituent ofS. rigidum root, falcarinol.