Antioxidant and Protective Effects of the Polyphenolic Glycoconjugate from Agrimonia eupatoria L. Herb in the Prevention of Inflammation in Human Cells.

Herein, structural and biological studies of a complex biopolymer (polyphenolic glycoconjugate) isolated from the flowering parts of Agrimonia eupatoria L. (AE) are presented. Spectroscopic analyses (UV-Vis and 1H NMR) of the aglycone component of AE confirmed that it consists mainly of aromatic and aliphatic structures characteristic of polyphenols. AE showed significant free radical elimination activity, i.e., ABTS+ and DPPH·, and was an effective copper reducing agent in the CUPRAC test, eventually proving that AE is a powerful antioxidant. AE was nontoxic to human lung adenocarcinoma cells (A549) and mouse fibroblasts (L929) and was nongenotoxic to S. typhimurium bacterial strains TA98 and TA100. Moreover, AE did not induce the release of proinflammatory cytokines such as interleukin 6 (IL-6) and tumor necrosis factor (TNF-α) by human pulmonary vein (HPVE-26) endothelial cells or human peripheral blood mononuclear cells (PBMCs). These findings correlated with the low activation of the transcription factor NF-κB in these cells, which plays an important role in the regulation of the expression of genes responsible for inflammatory mediator synthesis. The AE properties described here suggest that it may be useful for protecting cells from the adverse consequences of oxidative stress and could be valuable as a biomaterial for surface functionalization.

Effectivity of polyphenolic polysaccharide-proteins isolated from medicinal plants as potential cellular immune response modulators.

Traditional medicinal herbs as Echinacea purpurea and Erigeron canadensis are recommended as a complementary supplementation for the treatment of diseases associated with immunological inflammation (e.g. common cold, coughs, bronchitis, upper respiratory infections, immunodeficiencies). This pathologic conditions are accompanied by the wide range of malfunctions or imbalances of the immune system, thus there is increased necessity for search of novel immunomodulation trends and immunopharmacologically active phytosubstances for effective pharmaco-immunomodulatory therapy. Anti-inflammatory immunobiological activity of polyphenolic polysaccharide-proteins of Echinacea purpurea and Erigeron canadensis are still not studied. Our results demonstrated the immunobiological effectivity of selected herbal polyphenolic polysaccharide-proteins isolated from flowers of medicinal plants Echinacea purpurea and Erigeron canadensis resulting into the significant immunostimulation of inflammatory TNF-α, IL-6, IL-1ß and IL-12 cytokines (p < 0.001). Both herbal polyphenolic polysaccharide-proteins triggered cell release of anti-inflammatory interleukin IL-10 (p < 0.001). Furthermore, the inductive cell release of growth factors M-CSF and GM-CSF has been demonstrated (p < 0.001). E. purpurea and E. canadensis polyphenolic polysaccharide-proteins accelerated the efficacy of cellular phagocytosis and free radical release, more pronounced with Erigeron treatment.

Chemical characteristics and significant antitussive effect of the Erigeron canadensis polyphenolic polysaccharide-protein complex.

ETHNOPHARMACOLOGICAL RELEVANCE: Erigeron canadensis has been used in traditional medicine to treat a variety of respiratory diseases, including acute upper and lower respiratory tract infections and cough-related asthma. There is as yet no relevant experimental or clinical study in the scientific literature evaluating the efficacy of plants in these disorders. AIM OF THE STUDY: To investigate the active ingredients in Erigeron canadensis, a complex isolated from flowering parts of a plant was tested for airway defense reflexes, in particular for cough reflexes and airway reactivity. Both were experimentally induced by a chemical irritant that simulated the inflammatory conditions of their formation. MATERIAL AND METHODS: The polyphenolic polysaccharide-protein (PPP) complex was isolated from the flowering parts of Erigeron canadensis by hot alkaline extraction and a multi-stage purification process. The antitussive activity was confirmed as a decrease in the number of citric acid-induced coughs and the bronchodilator effect was verified as a decrease in specific airway resistance (sRaw) in conscious guinea pigs. RESULTS: The dark brown Erigeron complex with a molecular weight of 38,000 g/mol contained phenolics (13.2% wt%), proteins (16.3% wt%), and uronic acids (6.3% wt%). The neutral carbohydrate part of Erigeron consisted mainly of xylose (12.1 wt%), glucose (13.3 wt%), arabinose (24.1 wt%), and galactose (41.0 wt%) residues. Arabinogalactan and 4-OMe-glucuronoxylan have been found to be the major polysaccharides in the Erigeron complex. Using a method of chemically-induced cough reflex and guinea pigs test system the Erigeron complex exhibited statistically significant, the dose-dependent antitussive activity, which was similar to that of the centrally-acting opioid agonist codeine. CONCLUSION: Pharmacological tests have revealed a new pharmacodynamic effect of the Erigeron complex, namely an antitussive effect. Its activity was most pronounced in comparison with all previously tested compounds from other medicinal plants and approached the effect of codeine, the most potent antitussive used in clinical practice. The results provide the scientific basis for the application of this herb in traditional medicine.

Studying the "Rigid-Flexible" Properties of Polymeric Micelle Core-Forming Segments with a Hydrophobic Phthalocyanine Probe Using NMR and UV Spectroscopy.

The aim of the performed studies was to thoroughly examine the internal structure of self-assembled nanocarriers (i.e., polymeric micelles-PMs) by means of a hydrophobic phthalocyanine probe in order to identify the crucial features that are required to enhance the photoactive probe stability and reactivity. PMs of hydrophilic poly(ethylene glycol) and hydrophobic poly(ε-caprolactone) (PCL) or poly(d,l-lactide) (PDLLA) were fabricated and loaded with tetra tert-butyl zinc(II) phthalocyanine (ZnPc-t-but4), a multifunctional spectroscopic probe with a profound ability to generate singlet oxygen upon irradiation. The presence of subdomains, comprising "rigid" and "flexible" regions, in the studied block copolymers' micelles as well as their interactions with the probe molecules, were assessed by various high-resolution NMR measurements [e.g., through-space magnetic interactions by the 1D NOE effect, pulsed field gradient spin-echo, and spin-lattice relaxation time (T1) techniques]. The studies of the impact of the core-type microenvironment on the ZnPc-t-but4 photochemical performance also included photobleaching and reactive oxygen species measurements. ZnPc-t-but4 molecules were found to exhibit spatial proximity effects with both (PCL and PDLLA) hydrophobic polymer chains and interact with both subdomains, which are characterized by different rigidities. It was deduced that the interfaces between particular subdomains constitute an optimal host space for probe molecules, especially in the context of photochemical stability, photoactivity (i.e., for significant enhancement of singlet oxygen generation rates), and aggregation prevention. The present contribution proves that the combination of an appropriate probe, high-resolution NMR techniques, and UV-vis spectroscopy enables one to gain complex information about the subtle structure of PMs essential for their application as nanocarriers for photoactive compounds, for example, in photodynamic therapy, nanotheranostics, combination therapy, or photocatalysis, where the micelles constitute the optimal microenvironment for the desired photoreactions.

Optimization of Ultrasound-Assisted Extraction of Functional Food Fiber from Canadian Horseweed (Erigeron canadensis L.).

Much attention has been recently paid to the design of sustainable processes for the production of functional food additives based on renewable resources. Thus, methods for incorporation of green techniques in treatment of undeveloped biomass, resulting in value-added bioproducts, are in great demand. We focus here on the biological activity and chemical properties of Erigeron canadensis (horseweed) functional food fiber, which can be strongly affected by the extraction procedure employed. In the present contribution, we report on an attempt to introduce a sustainable and energy-efficient ultrasound-assisted extraction process, followed by a multistep purification procedure, resulting in a macromolecular plant-derived anticoagulant agent. The most efficient ultrasound-assisted process was determined by optimization through the response surface methodology I-optimal design (24). A comparison with the conventional procedure for retrieval of horseweed biomacromolecules revealed that the optimized ultrasound-assisted extraction was more sustainable, with the cumulative energy demand being 38% lower (12.2 MJ), 6.6 times reduced water consumption (3.5 L), and 1.2 times shorter (41 h) total processing time. Moreover, the optimal ultrasound-assisted extraction process-purified food fiber turned out to be a better anticoagulant agent by 57%, compared to a conventional product, and was a more selective indirect inhibitor of the human Xa coagulation factor.

Polish Propolis-Chemical Composition and Biological Effects in Tongue Cancer Cells and Macrophages.

The purpose of this study was to compare the chemical composition and biological properties of Polish propolis. Ethanol, ethanol-hexane, hexane and hexane-ethanol extracts of propolis from three different regions of Poland were prepared. On the basis of the evaluation of their chemical composition as well as the extraction yield and free radical scavenging activity, the ethanol and hexane-ethanol extractions were proposed as the most effective methods. Subsequently, the biological properties of the extracts were evaluated to investigate the selectivity of an anticancer effect on tongue cancer cells in comparison to normal gingival fibroblasts. The obtained products demonstrated anticancer activity against tongue cancer cells. Additionally, when the lowest extract concentration (100 µg/mL) was applied, they were not cytotoxic to gingival fibroblasts. Finally, a possible anti-inflammatory potential of the prepared products was revealed, as reduced mitochondrial activity and proliferation of macrophages exposed to the extracts were observed. The results obtained indicate a potential of Polish propolis as a natural product with cancer-selective toxicity and anti-inflammatory effect. However, further studies are still needed to thoroughly explain the molecular mechanisms of its action and to obtain the promising health benefits of this versatile natural product.

Polyphenolic-polysaccharide conjugates from medicinal plants of Rosaceae/Asteraceae family protect human lymphocytes but not myeloid leukemia K562 cells against radiation-induced death.

The aim of the study was to investigate whether the polyphenolic-polysaccharide conjugates (PPCs), isolated from flowers of Sanguisorba officinalis L. and Erigeron canadensis L., and from leaves of Fragaria vesca L. and Rubus plicatus Whe. Et N. E., can protect human peripheral blood mononuclear cells (PBMCs) against gamma-irradiation damage while maintaining the radiosensitivity of the myeloid leukemia K562 cell line. PPCs isolated from the four plant sources are water-soluble macromolecules (14-50 kDa) that were previously chemically and structurally characterized. Cells were incubated with PPCs (25 µg/ml, 1 h) prior exposure to 15 Gy gamma-irradiation, non-irradiated appropriate samples served as controls. It was found that the PPCs were able to increase the post-radiation viability of PBMCs by inhibiting apoptosis, while they did not protect the leukemic cells against radiation-induced apoptotic death. The PPCs offered an efficient protection of PBMCs through scavenging of intracellular ROS and decreasing DNA damage, while they provided no reduction of the oxidative stress and DNA damage in K562 cells. Our findings strongly suggest that the PPCs, especially these isolated from S. officinalis and E. canadensis, can selectively protect normal lymphocytes against radiation injury, therefore they meet the criteria of radioprotectors for potential use in radiotherapy.

The polyphenolic-polysaccharide complex of Agrimonia eupatoria L. as an indirect thrombin inhibitor - isolation and chemical characterization.

The polyphenolic-polysaccharide complex was isolated from the dried aerial parts of the medicinal plant Agrimonia eupatoria L. using a multi-step process involving the degreasing of the plant material by extraction with organic solvents, followed by extraction with hot alkali, neutralization, further separation with organic solvents and dialysis. The complex was homogeneous with a molecular weight of about 55 × 103 g/mol and consisted mainly of carbohydrates and polyphenols matrix, composed of lignin-related units, with the dominance of dimethoxyphenyl structures. The carbohydrate moiety consists mostly of arabinogalactan associated with highly esterified rhamnogalacturonan. In vitro anticoagulant studies revealed the ability of the A. eupatoria complex to inhibit plasma clot formation, mainly in the intrinsic pathway of the blood coagulation cascade. Further studies on the mechanisms of this anticoagulant activity revealed that the isolate was primarily an indirect inhibitor of thrombin, mediated by antithrombin or by heparin cofactor II. Such mechanism of action is characteristic for highly sulfated glycosaminoglycans.

Enhancing biogas plant production using pig manure and corn silage by adding wheat straw processed with liquid hot water and steam explosion.

BACKGROUND: Pig manure utilization and valorization is an important topic with tightening regulations focused on ecological and safety issues. By itself pig manure is a poor substrate for biogas production because of its excessive nitrogen content relative to available organic carbon. Such substrate is alkaline, and methanogenesis can be suppressed, and so additional substrates with high organic carbon must be added. The most promising is straw, which is available from adjacent biogas plant cultures. However, the abundant lignocellulosic biomass of wheat straw undergoes slow decomposition, and only a fraction of the chemical energy can be converted into biogas; thus economical methods for pretreatment increasing bioavailability are sought. RESULTS: A method was investigated to increase the methane yield in a full-scale plant for co-fermenting pig manure with corn silage, which was the default substrate in the original source reactors. Increased lignocellulosic bioavailability of wheat straw was achieved by combining liquid hot water (LHW) and steam explosion (SE). According to FT-IR analysis, the treatment resulted in hemicellulose hydrolysis, partial cellulose depolymerization, and lignin bond destruction. Low-mass polysaccharides (0.6 × 103 g mol-1) had significantly higher concentration in the leachate of LHW-SE wheat straw than raw wheat straw. The methanogenic potential was evaluated using inoculum from two different biogas plants to study the influence of microorganism consortia. The yield was 24-34% higher after the pretreatment process. In a full-scale biogas plant, the optimal conditions were ~ 165 °C, ~ 2.33 MPa, and 10 min in LHW and ~ 65 °C and ~ 0.1 MPa for SE. The processes did not generate detectable inhibitors according to GC-MS analysis, such as furfural and 5-hydroxymethylfurfural. CONCLUSIONS: The LHW-SE combined pretreatment process increases the bioavailability of carbohydrates from wheat straw. The LHW-SE treated wheat straw gave similar biogas yields to corn silage, thus enables at least partial replacement of corn silage and is good for diversification of substrates. Surprisingly, microorganisms consortia from other biogas plant fed with other substrates may have higher efficiency in utilization of tested substrate. Thus, methanogenic consortia may be considered in the process of optimization at industrial scale. The efficiency was calculated, and the LHW-SE may be profitable at full industrial scale and further optimization is proposed.

A comparative study on the radioprotective potential of the polyphenolic glycoconjugates from medicinal plants of Rosaceae and Asteraceae families versus their aglycones.

Radioprotective potential of the polyphenolic glycoconjugates, isolated from flowers of Sanguisorba officinalis L. (So) and Erigeron canadensis L. (Ec), and from leaves of Fragaria vesca L. (Fv) and Rubus plicatus Whe. Et N. E. (Rp) as well as their aglycones (SoA, EcA, FvA and RpA, respectively), against γ-radiation-induced lipid peroxidation in human plasma and DNA damage in lymphocytes, were investigated in vitro. These properties were assessed by measuring the concentration of thiobarbituric acid reactive substances (TBARS) and using the alkaline comet assay, and were compared to the protective effects of rutin (R) and quercetin (Q). Cytotoxicity of the glycoconjugates/aglycones towards L929 mouse fibroblasts and human lymphocytes were also measured. Plant products from S. officinalis, similar to Q, were able to reduce the most radiation-induced lipid peroxidation as well as DNA damage and extent of oxidative damage to the DNA basis. Contrary to the pure flavonoids, where Q was shown to be significantly more effective than its glycoside R, the results did not show more benefit with application of SoA/EcA over So/Ec in terms of lipid peroxidation inhibition. Moreover, glycoconjugates Ec and So showed much higher capacity in protecting lymphocytes against radiation-induced genotoxicity which may suggest that between the polyphenolic and polysaccharide parts exist some synergistic effects. There were no significant differences between Fv versus FvA or Rp versus RpA in terms of the provided radioprotection. Summarizing, plant glycoconjugates isolated by the multi-step method offered sufficient radioprotection. In addition, they possess many advantages, compared to the synthetic polyphenolic compounds or the plant extracts, such as water-solubility and minor toxicity.

Polyphenolic glycoconjugates from medical plants of Rosaceae/Asteraceae family protect human lymphocytes against γ-radiation-induced damage.

Radioprotective effects of the water-soluble polyphenolic glycoconjugates, isolated from flowers of Sanguisorba officinalis L.(SO) and Erigeron canadensis L.(EC), and from leaves of Fragaria vesca L. (FV) and Rubus plicatus Whe. Et N. E. (RP), against γ-radiation-induced toxicity in human peripheral blood lymphocytes were investigated. Cell treatment with glycoconjugates (1, 5 and 25µg/mL) prior exposure to 10/15Gy radiation resulted in concentration-dependent reduction of DNA damage including oxidative DNA lesions (comet assay), substantial inhibition of lipid peroxidation (TBARS) and restoration of superoxide dismutase and S-glutathione transferase activities. Glycoconjugates isolated from SO and EC ensured better protection versus these from RP and FV, with the SO product potential comparable to that of the reference quercetin. Strong antioxidant/radioprotective activity of the SO and EC glycoconjugates could be attributed to high abundance of syringol-type and ferulic acid units in their matrices, respectively. Moreover, polyphenolic glycoconjugates (25µg/mL), including RP and FV products, significantly decreased DNA damage when applied post-radiation suggesting their modulating effects on DNA repair pathways. Preliminary data on the glycoconjugate phenolic structural units, based on GLC/MS of the products of pyrolysis and in situ methylation, in relation to application of plant products as potential radioprotectors is promising and deserves further investigation.

Polyphenolic-polysaccharide conjugates of Sanguisorba officinalis L. with anticoagulant activity mediated mainly by heparin cofactor II.

A macromolecular complex has been isolated from the dried flowering parts of medicinal plant Sanguisorba officinalis L. (So) by multi-step extraction procedure, including that with extraction by organic solvents to degrease the plant material, then with hot alkali, followed by neutralization, partitioning with organic solvents and dialysis. The complex was purified by size-exclusion chromatography into five fractions labeled as So1-So5. Individual fractions differed in the chemical composition and molecular weight distribution patterns. In vitro anticoagulant activity tests showed in all fractions more or less important inhibition of plasma clots, however, So3 and So4 were the most active. The anticoagulant activity of So3 was even more significant than that of the unfractionated complex So. These S. officinalis conjugates were able to inhibit mainly the activity of thrombin when they were mediated by heparin cofactor II, but what was unexpected they were the non-direct inhibitors of factor Xa, mediated by antitrombin, where such mechanism of action is typical for a highly sulphated glycosaminoglycans.

Polyphenolic-polysaccharide conjugates from plants of Rosaceae/Asteraceae family as potential radioprotectors.

Polyphenolic-polysaccharide macromolecular, water-soluble glycoconjugates, isolated from the selected medicinal plants of Rosaceae/Asteraceae family: from leaves of Fragaria vesca L., Rubus plicatus Whe. et N. E., and from flowering parts of Sanguisorba officinalis L., and Erigeron canadensis L., were investigated for their ability to protect proteins and lipids of human plasma against γ-radiation-induced oxidative damage. Treatment of plasma with plant conjugates (6, 30, 150 µg/ml) prior exposure to 100 Gy radiation resulted in a significant inhibition of lipid peroxidation, evaluated by TBARS levels; conjugates isolated from E. canadensis and R. plicatus and a reference flavonoid quercetin showed similar high potential (approx. 70% inhibition, at 6 µg/ml). The conjugates prevented radiation-induced oxidation of protein thiols and significantly improved plasma total antioxidant capacity, estimated with Ellman's reagent and ABTS(.+) assay, respectively. The results demonstrate by the first time a significant radioprotective capability of the polyphenolic-polysaccharide conjugates isolated from E. canadensis, R. plicatus, S. officinalis and to the less extent from F. vesca. The abilities of these substances to inhibit radiation-induced lipid peroxidation and thiol oxidation in plasma seems to be mediated, but not limited to ROS scavenging activity.

Echinacea complex--chemical view and anti-asthmatic profile.

ETHNOPHARMACOLOGICAL RELEVANCE: Echinacea purpurea (L.) Moench is one of the mostly used herbs in the traditional medicine for the treatment of respiratory diseases. Modern interest in Echinacea is directed to its immunomodulatory activity. Recent studies have shown that secretion of asthma-related cytokines in the bronchial epithelial cells can be reversed by Echinacea preparations. AIM OF THE STUDY: To examine the pharmacodynamics profile of Echinacea active principles, a complex has been isolated from its flowers by alkaline extraction and has been tested using an animal model of allergic asthma. MATERIAL AND METHODS: The structural features of Echinacea purpurea complex was determined using chemical and spectroscopic methods. Allergic inflammation of the airways was induced by repetitive exposure of guinea pigs to ovalbumin. Echinacea complex was then administered 14 days in 50mg/kg b.w. daily dose perorally. Bronchodilatory effect was verified as decrease in the specific airway resistance (sRaw) in vivo and by reduced contraction amplitude (mN) of tracheal and pulmonary smooth muscle to cumulative concentrations of acetylcholine and histamine in vitro. The impact on mucociliary clearance evaluated measurement of ciliary beat frequency (CBF) in vitro using LabVIEW™ Software. Anti-inflammatory effect of Echinacea complex was verified by changes in exhaled NO levels and by Bio-Plex® assay of Th2 cytokine concentrations (IL-4, IL-5, IL-13 and TNF-alpha) in serum and bronchoalveolar lavage fluid (BALF). RESULTS: Chemical and spectroscopic studies confirmed the presence of carbohydrates, phenolic compounds and proteins, as well as the dominance of rhamnogalacturonan and arabinogalactan moieties in Echinacea complex. The significant decrease in sRaw values and suppressed histamine and acetylcholine-induced contractile amplitude of isolated airways smooth muscle that were similar to effects of control drug salbutamol confirmed Echinacea complex bronchodilatory activity. The anti-inflammatory effect was comparable with that of control agent budesonide and was verified as significantly reduced exhaled NO levels and concentration of Th2 cytokines in serum and BALF. The values of CBF were changed only insignificantly on long-term administration of Echinacea complex suggested its minimal negative impact on mucociliary clearance. CONCLUSION: Pharmacodynamic studies have confirmed significant bronchodilatory and anti-inflammatory effects of Echinacea complex that was similar to effects of classic synthetic drugs. Thus, results provide a scientific basis for the application of this herb in traditional medicine as a supplementary treatment of allergic disorders of the airways, such as asthma.

Chemical and pharmacological profiles of Echinacea complex.

Echinacea purpurea has a long history in traditional medicine. To verify the pharmacological efficacy of active principles, a polysaccharide-phenolic-protein complex has been isolated from flowering parts of herb by alkaline extraction. It showed on GPC and HPLC one peak of molecular mass around 10 kDa. Chemical and spectroscopic analyses revealed carbohydrate, phenolic and protein contents in Echinacea complex. Pharmacological tests have shown its marked cough suppressing and bronchodilatory effects. The antitussive effect of Echinacea was similar to the narcotic drug codeine and the bronchodilatory effect was more significant than salbutamol, the antiasthmatic drug used in a clinical practice. Pharmacodynamic study shows the beneficial effects of Echinacea complex on the respiratory system and highlights the great potential for development of antitussive and bronchodilatory drugs from natural sources.

Metabonomic analysis of serum of workers occupationally exposed to arsenic, cadmium and lead for biomarker research: a preliminary study.

Environmental metabonomics is the application of metabonomics to characterize the interactions of organisms with their environment. Metabolic profiling is an exciting addition to the armory of the epidemiologist for the discovery of new disease risk biomarkers and diagnostics. This work is a continuation of research searching for preclinical serum markers in a group of 389 healthy smelter workers exposed to lead, cadmium and arsenic. Changes in the metabolic profiles were studied using Proton Nuclear Magnetic Resonance Spectroscopy on pooled serum samples from both the metal exposed and control groups. These multivariate metabonomic datasets were analyzed with Principal Component Analysis and Partial Least Squares Discriminant Analysis. Analysis of metabolic profiles of people exposed to heavy metals suggests energy metabolism disturbance induced by heavy metals. Changes in lipid fraction (very-low-density lipoprotein - VLDL, low-density lipoprotein - LDL), unsaturated lipids and in the level of amino acids suggest perturbation of the metabolism of lipids and amino acids. This study illustrated the high reliability of NMR-based metabonomic profiling on the study of the biochemical effects induced by the mixture of heavy metals. This approach is capable of identifying intermediate biomarkers of response to toxicants at environmental/occupational concentrations, paving the way to its use in a monitoring of smelter workers exposed to low doses of lead, cadmium and arsenic.

[Biosensors as a tool to use in the experiments on blood and selected blood proteins]. / Biosensory jako narzedzie do wykorzystania w badaniach krwi oraz wybranych bialek krwi.

Over last few years, biosensors have become increasingly used as a research tool. Using innovative techniques of detection and estimation of the strength of intermolecular bonds, is particularly important in biochemical processes, including the study of mechanisms of interactions between plasma proteins in the circulatory system. With the numerous enhancements biosensors have become extremely sensitive devices, allowing for analysis of diverse biological material. Moreover, the use of immobilization techniques enables to use sample repeatedly, which significantly reduces costs and the ability to monitor tests in real-time shorten the time of experiment. The presented work discusses examples of the usage of biosensors in the research on the mechanisms of the interactions of blood plasma proteins. The experiments on cancer biomarkers present in the blood circulation system, protein C deficiency causing non-controlled hemorrhagic accidents, and on the level of fibrinogen, as well as 20S proteasome concentration in plasma, are just some examples of biosensors usage in the analyses of blood. They are also applicable in the research work performed the project WROVASC--Integrated Cardiovascular Center, concerning the mechanisms of anticoagulant activity in blood plasma of the polyphenolic-polysaccharide macromolecules of plant origin.

[The plant anticoagulants with perspective to use in treatment of thrombosis]. / Antykoagulanty pochodzenia roslinnego z perspektywa wykorzystania w leczeniu zakrzepic.

The diseases of blood circulation system--cardiovascular diseases are the main causes of mortality in developing, low and middle-income countries all over the world. The specialists recommend the prophylaxis to avoid the dangerous complications connected with these diseases, what can reduce the final treatment costs. All over the world there is continuous research of novel, therapeutically better, more effective anticoagulant or anti-platelet agents, with multiple targets, without so many side effects. Plant material is a good source to do this kind of research. The authors show the results of their few years research on polyphenolic-polysaccharide plant conjugates, isolated from medicinal plants, popular in Poland, which is continuing in the framework of the project WROVASC--Integrated Center of Cardiovascular Medicine. This research group has been working on isolation, structure characterization and biological activity of these macromolecular compounds. Because of anticoagulant, antioxidant as well as anti-platelet properties of these plant structures they are promising to be a new source of the innovative therapeutic agent.

Assessment of extracts of Helichrysum arenarium, Crataegus monogyna, Sambucus nigra in photoprotective UVA and UVB; photostability in cosmetic emulsions.

The aim of our study was to investigate the photoprotective activity and photostability efficacy of sunscreen formulations containing Helichrysum arenarium, Sambucus nigra, Crataegus monogyna extracts and their combination. UV transmission of the emulsion films was performed by using diffuse transmittance measurements coupling to an integrating sphere. In vitro photoprotection and photostability efficacy were evaluated according to the following parameters: sun protection factor (SPF), UVA protection factor (PF-UVA), UVA/UVB ratio and critical wavelength (λc) before and after UV irradiation. The results obtained show that the formulations containing polyphenols fulfill the official requirements for sunscreen products due to their broad spectrum of UV protection combined with their high photostability and remarkable antioxidant properties. Therefore H. arenarium, S. nigra, C. monogyna extracts represent useful additives for cosmetic formulation.