Comparison of antioxidant and antimicrobial activities of extracts obtained from Salvia glutinosa L. and Salvia officinalis L.

Antioxidant and antimicrobial activities as well as total phenols and flavonoids contents of Salvia glutinosa L. (glutinous sage) and Salvia officinalis L. (sage) extracts were studied. Methanol and aqueous ethanol (70% v/v) were used for extraction of bioactive compounds, both in the presence and the absence of ultrasound, from herb and the spent plant material remaining after the essential oil hydrodistillation. The ratio of plant material to extracting solvent was 1:10 g/ml. Antioxidant and antimicrobial activities of the extracts were found to depend on the type of plant material and the extraction conditions. The plant materials from which essential oil had been recovered were proven to be valuable raw materials for making various herbal preparations

Antioxidant activity, the content of total phenols and flavonoids in the ethanol extracts of Mentha longifolia (L.) Hudson dried by the use of different techniques

In this study, we have examined the yield of extracted substances obtained by means of extraction using 70 % ethanol (v/v), the content of total phenols and flavonoids, as well as the antioxidant activity of the extracts obtained from the samples of the herbs dried by means of different techniques. Wild mint Mentha longifolia (L.) Hudson was dried naturally in a laboratory oven at a temperature of 45 °C and in an absorptive low temperature condensation oven at 35°C. The highest yield of extracts was obtained from the naturally dried herbs and the lowest from the herbs dried in the low temperature condensation drying oven. The content of total phenols and flavonoids was determined by spectrophotometric methods with an FC reagent and by the complexation reaction with aluminium-chloride, respectively. The extract of the naturally dried herbs had the highest overall content of phenols (113.8±2.0 mg of gallic acid/g of the dry extract) and flavonoids (106.7±0.3 mg of rutin/g of the dry extract). The highest antioxidant activity determined by the FRAP and DPPH assay was determined in the extracts obtained from naturally dried herbs (2.76±0.15 mmol Fe2+/mg of the dry extract and EC50=0.022±0.001 mg/ml), while the lowest was obtained from the extracts of herbs dried in the laboratory oven (1.13±0.11 mmol Fe2+/mg of the dry extract and EC50=0.033±0.001 mg/ml). The HPLC-DAD analysis result show that the greatest content of phenolic compounds show extract obtained from naturally dried plant material. The dominant phenolic component in the all extracts is Kaempferol 3-O-glucoside. The content of all phenolic compound strongly depend on the drying conditions

Direct ultrasound-assisted extraction and characterization of phenolic compounds from fresh houseleek (Sempervivum marmoreum L.) leaves

The effects of ultrasound power and frequency on the yield of total extractive substances (TES), total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity (AOA) of fresh houseleek leaves extracts obtained by direct ultrasound-assisted extraction (DUAE) were studied. Preliminary extraction of plant material was performed using methanol, acetone and 2-propanol by Soxhlet extraction. It was found that maximum TES yield could be obtained by methanol extraction (2.91±0.02), followed by acetone and 2-propanol with a TES yield of 2.32±0.01 and 2.01±0.03 g per 100 g of fresh plant material, respectively. In the fresh houseleek leaves extracts obtained by DUAE and methanol as the chosen solvent, TPC, TFC and AOA were in the ranges of: 40.5–85.9 mg gallic acid/g dry extract, 12.7–19.3 mg rutin/g dry extract and 24.6–108.2μg/ml, respectively. The results showed that the increase in the ultrasound power and extraction time have positive and significant (p < 0.05) effects on the TPC, TFC and AOA, while the increase in the ultrasound frequency leads to a decrease in the TPC, TFC and AOA of the extracts. A chromatographic analysis of crude extract identified the following: kaempferol 3-O-(6’’-O-malonylglucoside)- 7-O-glucosyde, kaempferol 3-O-glucoside-7-O-rhamnoside, luteolin 5-O-(6’’-O-malonylglucoside), kaempferol 3-O-(6’’-O-acetylglucoside)-7-O-rhamnoside, genkwanin 5-O-glucoside, luteolin 5-O-(6’’-O-malonylglucoside), kaempferol 3-O-(6’’-O-malonylglucoside), kaempferol 3-O-rhamnoside, quercetin, genkwanin 4’-O-glucoside and hyperoside. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 172047

Role of molecular mimicry and polyclonal cell activation in the induction of pathogenic beta 2-glycoprotein I-directed immune response in Balb/c mice upon hyperimmunization with tetanus toxoid

It is known that tetanus toxoid (TTd)-hyperimmunization induces increased titer of sera beta 2-glycoprotein I (beta 2GPI)-specific antibodies (Abs) in Balb/c mice. The concentrations of such induced anti-beta 2GPI Abs as well as their pathogenic potential are strongly influenced by the context of TTd application. beta 2GPI-specific immune response is established as a part of TTd-specific immune response by molecular mimicry mechanism due to structural homology between TTd and beta 2GPI. This finding is supported by the following facts: (1) cross-reactive Abs that recognize both TTd and beta 2GPI epitopes are present in Balb/c mice sera; (2) anti-TTd Abs secretion in splenic cultures is induced after beta 2GPI stimulation and vice versa. However, analyses of (1) IL-10 production following in vitro stimulation of immunized Balb/c mice splenocytes by TTd, beta 2GPI or glutaraldehyde-treated beta 2GPI and (2) specific impact of ConA and agonists of TLR2, TLR4, and TLR9 on anti-TTd and autoreactive Abs secretion strongly imply that these two branches of the TTd-induced immune response do not use identical cell populations and are regulated in a different way. Results presented in this paper describe that structural homology between foreign and self-antigens could focus mounted autoreactive immune response toward specific self-structure, but the context of antigen application, including a history of previous immune stimulations and adjuvants applied together with the antigen, are the main factors which determine the outcome of the induced immune response

Phenotypic and functional characteristics of splenocytes in tetanus toxoid-hyperimmunized Balb/c mice is influenced by the context of tetanus toxoid application

Purpose/Objective: The hyperimmunization with tetanus toxoid (TTd) induces protective TTd-specific as well as autoreactive b2-glycoprotein I (b2GPI)-specific immune responses in BALB/c mice. The overall immune response characteristics, especially its pathogenic potential, depended on adjuvants applied prior and in combination with TTd. Beside structural homology between TTd and b2GPI, tolerance toward b2GPI could be impaired by adjuvants acting as polyclonal stimulators. In order to clarify the impact of adjuvants, phenotypic and functional analyses of immune system cells within spleen were done upon immunization completion. Materials and methods: Non- or CFA-pretreated BALB/c mice were immunized with TTd (3 · 100 lg/dose; 2-week intervals) mixed with alum or 2.5M glycerol. Ex vivo analyses of CD3, CD4, CD8, CD19, CD 25, CD27 and mIgM expression on age-matched control and immunized mice’s splenocytes were done by flow cytometry. Changes in TLR2, TLR4 and TLR9 expression were assessed indirectly, by measuring cytokine production, following in vitro stimulation of splenocytes with appropriate agonist. Results: TTd-immunization diminished CD27 expression on T cells implying on their differentiation into potent effector cells. T cell activation (increase in CD25 expression and the raise of percentage of CD4+ CD8+ CD3+ ) and B cell activation (rise in percentage of CD19+ CD25+ cells and the increase of mIgM density) occurred in all immunized mice, being more intensive in CFA-pretreated groups. Irrespective to the applied immunization protocol, statistically significant rise in abundance of CD4- CD8- cells (often cited as cells having suppressive potential) within T cell pool was registered too. Differences in cytokines production (IL4, IL10, IFNc) registered upon in vitro stimulation with peptidoglycan, LPS and CpG ODN implied on context-dependant modulation of TLR2, TLR4 and TLR9 expression on splenocytes. Conclusions: TTd-hyperimmunization promoted concomitant rise in abundance of activated cells and the cells that have suppressive potential. This could be regarded as an attempt of the system to retain control. Imbalance in percentages and activities between activated cells and those having suppressive potential, highly influenced by the context of TTd application, is most likely the cause for the observed pathology appearance after TTd hyperimmunization

JSCS–3742 Original scientific paper

Comparative screening of the anti-oxidant and antimicrobial activities of Sempervivum marmoreum L. extracts obtained by various extraction technique