Qualitative and Quantitative Analysis of Secondary Metabolites in Morphological Parts of Paulownia Clon In Vitro 112® and Their Anticoagulant Properties in Whole Human Blood.

It is not easy to find data in the scientific literature on the quantitative content of individual phytochemicals. It is possible to find groups of compounds and even individual compounds rather easily, but it is not known what their concentration is in cultivated or wild plants. Therefore, the subject of this study was to determine the content of individual compounds in the new Paulownia species, Oxytree, developed in a biotechnology laboratory in 2008 at La Mancha University in Spain. Six secondary metabolites were isolated, and their chemical structure was confirmed by spectral methods. An analytical method was developed, which was then used to determine the content of individual compounds in leaves, twigs, flowers and fruits of Paulownia Clon in Vitro 112®. No flavonoids were found in twigs and fruits of Oxytree, while the highest phenylethanoid glycosides were found in twigs. In this study, we also focused on biological properties (anticoagulant or procoagulant) of extract and four fractions (A-D) of different chemical composition from Paulownia Clon in Vitro 112 leaves using whole human blood. These properties were determined based on the thrombus-formation analysis system (T-TAS), which imitates in vivo conditions to assess whole blood thrombogenecity. We observed that three fractions (A, C and D) from leaves decrease AUC10 measured by T-TAS. In addition, fraction D rich in triterpenoids showed the strongest anticoagulant activity. However, in order to clarify the exact mechanism of action of the active substances present in this plant, studies closer to physiological conditions, i.e., in vivo studies, should be performed, which will also allow to determine the effects of their long-term effects.

Giffonins, Antioxidant Diarylheptanoids from Corylus avellana, and Their Ability to Prevent Oxidative Changes in Human Plasma Proteins.

With the aim to explore the ability of diarylheptanoids to reduce oxidative changes in human plasma proteins, a phytochemical investigation of the MeOH extract of Corylus avellana leaves was perfomed. Analysis by LC-ESI/LTQOrbitrap/MS/MSn guided the isolation of two new diarylheptanoid derivatives, giffonins W (1) and X (2). The structures 1 and 2 were assigned by analysis of NMR data combined with a QM (quantum mechanical)/NMR approach. The absolute configurations of 1 and 2 were established by analysis of electronic circular dichroism (ECD) spectra compared with the TDDFT-simulated curves. The antioxidant activity of the new and known giffonins was evaluated by inhibition of human plasma lipid peroxidation. Giffonins with the highest inhibitory activity were tested for their ability to reduce oxidation of thiol groups and carbonylation in plasma proteins, and some of them exhibited higher antioxidant activity than curcumin.

Electrospray ionization mass spectrometry characterization of ubiquitous minor lipids and oligosaccharides in milk of the camel (Camelus dromedarius) and their inhibition of oxidative stress in human plasma.

The aim of this study was to characterize minor lipids in methanol fraction extracted from raw camel milk after loading it on a water-preconditioned short C18 open column and fractionating with a gradient of methanol/water. The C18 column showed high fractionation efficiency of minor lipids, such as glycosphingolipids, lipopolysaccharides, or oligosaccharides, when compared with other constituents, in particular polysaccharides, proteins, and free fatty acids. Liquid chromatography electrospray ionization tandem mass spectrometry in negative ion mode was used to identify 21 new glycosphingolipids, lipopolysaccharides, and oligosaccharides. Electrospray ionization tandem mass spectrometry was qualified to provide relevant data for recognizing the molecular mass, glycosylation sequences, and structure of saccharide moieties for the revealed compounds. The sequence of combinations of one selected lipopolysaccharide, which was considered the backbone of the remaining lipopolysaccharides, was confirmed in a density functional theory study. The obtained results showed that the tested fraction is a rich source of glycosphingolipids, lipopolysaccharides, and oligosaccharides with antioxidant activity.

Biological properties of Elaeagnus rhamnoides (L.) A. Nelson twig and leaf extracts.

BACKGROUND: Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson, SBT) is a valuable plant because of its medical and therapeutic potential. Different bioactive compounds in SBT berries are of special interest to various researchers. However, not only sea buckthorn berries, but also leaves of this plant (both fresh and dried) contain a lot of nutrients and bioactive compounds, including phenolic compounds. The present study was carried out in order to investigate antioxidant and anticoagulant properties of sea buckthorn twig and leaf extracts (0.5-50 µg/mL) by using various in vitro models. Moreover, the aim of present experiments was to compare the biological activity of SBT leaf extract and SBT twig extract with selected berry extracts (a rich source of phenolic compounds): SBT berry extract (flavonoids being the dominant components), a commercial extract from the berries of Aronia melanocarpa (Aronox®), and a grape seed extract. METHODS: We determined the effect of plant extracts on the oxidative stress using selected markers of this process, i.e. the level of carbonyl groups in proteins. Additionally, we analysed the potential mechanism of modulation of hemostatic properties of human plasma (using selected coagulation times). RESULTS: SBT twig and leaf extracts were observed to exhibit an antioxidant activity against two strong biological oxidants: hydrogen peroxide (H2O2) and H2O2/Fe (the donor of hydroxyl radicals), which induced human plasma lipid peroxidation and protein carbonylation. Both extracts also showed anticoagulant properties. CONCLUSIONS: Our present results have demonstrated that extracts from different parts of SBT, especially berries and twigs, in comparison to well-known berries (aronia and grape), may also be viewed as a good source of active substances - antioxidants for pharmacological or cosmetic applications. Moreover, it is very important from an economic point of view to know that there is a possibility of obtaining phenolic compounds not only from the berries or leaves, but also from twigs, which constitute a production waste.

Anti-Platelet Properties of Phenolic Extracts from the Leaves and Twigs of Elaeagnus rhamnoides (L.) A. Nelson.

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a small tree or bush. It belongs to the Elaeagnaceae family, and has been used for many years in traditional medicine in both Europe and Asia. However, there is no data on the effect of sea buckthorn leaves and twigs on the properties of blood platelets. The aim of the study was to analyze the biological activity of phenolic extracts from leaves and twigs of sea buckthorn in blood platelets in vitro. Two sets of extracts were used: (1) phenolic compounds from twigs and (2) phenolic compounds from leaves. Their biological effects on human blood platelets were studied by blood platelet adhesion, platelet aggregation, arachidonic acid metabolism and the generation of superoxide anion. Cytotoxicity was also evaluated against platelets. The action of extracts from sea buckthorn twigs and leaves was compared to activities of the phenolic extract (a commercial product from the berries of Aronia melanocarpa (Aronox®) with antioxidative and antiplatelet properties. This study is the first to demonstrate that extracts from sea buckthorn leaves and twigs are a source of bioactive compounds which may be used for the prophylaxis and treatment of cardiovascular pathologies associated with blood platelet hyperactivity. Both leaf and twig extracts were found to display anti-platelet activity in vitro. Moreover, the twig extract (rich in proanthocyanidins) displayed better anti-platelet potential than the leaf extract or aronia extract.

Cyclic Diarylheptanoids from Corylus avellana Green Leafy Covers: Determination of Their Absolute Configurations and Evaluation of Their Antioxidant and Antimicrobial Activities.

The methanol extract of the leafy covers of Corylus avellana, source of the Italian PGI (protected geographical indication) product "Nocciola di Giffoni", afforded two new cyclic diarylheptanoids, giffonins T and U (2 and 3), along with two known cyclic diarylheptanoids, a quinic acid, flavonoid-, and citric acid derivatives. The structures of giffonins T and U were determined as highly hydroxylated cyclic diarylheptanoids by 1D and 2D NMR experiments. Their relative configurations were assigned by a combined quantum mechanical/NMR approach, comparing the experimental 13C/1H NMR chemical shift data and the related predicted values. The absolute configurations of carpinontriol B (1) and giffonins T and U (2 and 3) were assigned by comparison of their experimental electronic circular dichroism curves with the TDDFT-predicted curves. The ability of the compounds to inhibit the lipid peroxidation induced by H2O2 and H2O2/Fe2+ was determined by measuring the concentration of thiobarbituric acid reactive substances. Furthermore, the antimicrobial activity of the methanol extract of leafy covers of C. avellana and of the isolated compounds against the Gram-positive strains Bacillus cereus and Staphylococcus aureus and the Gram-negative strains Escherichia coli and Pseudomonas aeruginosa was evaluated. Carpinontriol B (1) and giffonin U (3) at 40 µg/disk caused the formation of zones of inhibition.

ANTIOXIDANT PROPERTIES OF METHANOLIC EXTRACTS FROM THE SHOOTS AND ROOTS OF pRi-TRANSFORMED PLANTS OF REHMANNIA GLUTINOSA LIBOSCH.

The antioxidant activity of methanolic extracts derived from shoots (HR-shoots) and roots (HR-roots) of pRi-transformed Rehmannia glutinosa plants were determined. The activity was indicated by the ability of the plant extracts to inhibit superoxide anion (O2(-·)) generation and thiobarbituric acid reactive substances (TBARS) production in resting blood platelets and platelets activated by thrombin. The strongest activity was exhibited by the HR-shoot extract (50 µg/mL). The present study also examines the antioxidant properties of the plant extracts against human plasma lipid peroxidation induced by strong biological oxidants: hydrogen peroxide (H2O2) and H2O2/Fe. The study shows that extracts from transformed R. glutinosa plants may be a promising source of natural antioxidants, which would be valuable in various cardiovascular diseases. The extracts may also protect lipids against oxidative modifications.

COMMERCIAL EXTRACT FROM ARONIA AS A MODULATOR OF ADHESIVE PROPERTIES OF FIBRINOGEN TREATED WITH HOMOCYSTEINE AND ITS THIOLACTONE IN VITRO.

Research has confirmed the positive effect of berries of Aivnia melanocarpa on the cardiovascular system. The protective effects of polyphenol-rich extract from berries of A. melanocarpa against changes in biological properties of fibrinogen were studied. In in vino model of hyperhomocysteinemia the capability of fibrinogen to interact with human blood platelets was measured by platelet adhesion in the presence of extract fromA. nelanocapa. We induced hyperhomocystenemia using a reduced form of homocysteine (Hey, at a final concentration of 0.01. 0.1 and 1 µM) and the most reactive form of Hey - its cyclic thioester, homocysteine thiolactone (HTL, at a final concentration of 0.1, 0.5 and I µM). It was observed that Hey or HTL-treated fibrinogen, in comparison with untreated molecule, had a distinct capability to mediate blood platelet adhesion. The experiments also indicate that polyphenol-rich extract from black chokeberries (at final concentrations of 2.5-10 pM/mL) reduced the toxic action of Hey and HTL on the adhesive properties of fibrinogen. The possible protection exerted by black chokeberry extract, through restoring the platelet adhesion of Hey or HTL treated fibrinogen, may be important for vascular diseases.

Hydrogen sulfide decreases the plasma lipid peroxidation induced by homocysteine and its thiolactone.

Hydrogen sulfide (H2S) has been investigated widely in recent years. H2S plays a variety of roles in different biological systems, including cardiovascular system. It is the final product of amino acids metabolism, which contains sulfur-cysteine and homocysteine (Hcy). In human plasma, there are several various forms of homocysteine: free Hcy, protein-bound Hcy (S-linked, and N-linked), and homocysteine thiolactone (HTL). Our previous works have shown that both Hcy in the reduced form and its thiolactone may modify fibrinolysis, coagulation process, and biological activity of blood platelets. Moreover, we have observed that HTL, like its precursor-Hcy stimulated the generation of superoxide anion radicals (O 2 (-•) ) in blood platelets. The aim of our study in vitro was to establish the influence of sodium hydrosulfide (NaHS, as a fast-releasing H2S donor; at tested concentrations: 10-1000 µM) on the plasma lipid peroxidation induced by the reduced Hcy (at final concentrations of 0.01-1 mM) and HTL (at final concentrations of 0.1-1 µM). Our results indicate that 10 and 100 µM NaHS decreased the lipid peroxidation in plasma treated with 1 mM Hcy or 1 µM HTL (when NaHS and Hcy/HTL were added to plasma together). The protective effect of 10 and 100 µM NaHS against the lipid peroxidation in plasma preincubated with 1 mM Hcy or 1 µM HTL was also observed. Considering the data presented in this study, we suggest that the lipid peroxidation (induced by different forms of homocysteine) may be reduced by hydrogen sulfide.

New Findings Regarding the Effects of Selected Blue Food Colorants (Genipin, Patent Blue V, and Brilliant Blue FCF) on the Hemostatic Properties of Blood Components In Vitro.

Natural and synthetic colorants present in food can modulate hemostasis, which includes the coagulation process and blood platelet activation. Some colorants have cardioprotective activity as well. However, the effect of genipin (a natural blue colorant) and synthetic blue colorants (including patent blue V and brilliant blue FCF) on hemostasis is not clear. In this study, we aimed to investigate the effects of three blue colorants-genipin, patent blue V, and brilliant blue FCF-on selected parameters of hemostasis in vitro. The anti- or pro-coagulant potential was assessed in human plasma by measuring the following coagulation times: thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (APTT). Moreover, we used the Total Thrombus formation Analysis System (T-TAS, PL-chip) to evaluate the anti-platelet potential of the colorants in whole blood. We also measured their effect on the adhesion of washed blood platelets to fibrinogen and collagen. Lastly, the cytotoxicity of the colorants against blood platelets was assessed based on the activity of extracellular lactate dehydrogenase (LDH). We observed that genipin (at all concentrations (1-200 µM)) did not have a significant effect on the coagulation times (PT, APTT, and TT). However, genipin at the highest concentration (200 µM) and patent blue V at the concentrations of 1 and 10 µM significantly prolonged the time of occlusion measured using the T-TAS, which demonstrated their anti-platelet activity. We also observed that genipin decreased the adhesion of platelets to fibrinogen and collagen. Only patent blue V and brilliant blue FCF significantly shortened the APTT (at the concentration of 10 µM) and TT (at concentrations of 1 and 10 µM), demonstrating pro-coagulant activity. These synthetic blue colorants also modulated the process of human blood platelet adhesion, stimulating the adhesion to fibrinogen and inhibiting the adhesion to collagen. The results demonstrate that genipin is not toxic. In addition, because of its ability to reduce blood platelet activation, genipin holds promise as a novel and valuable agent that improves the health of the cardiovascular system and reduces the risk of cardiovascular diseases. However, the mechanism of its anti-platelet activity remains unclear and requires further studies. Its in vivo activity and interaction with various anti-coagulant and anti-thrombotic drugs, including aspirin and its derivatives, should be examined as well.

Epicatechin inhibits human plasma lipid peroxidation caused by haloperidol in vitro.

Epicatechin belongs to flavonoids protecting cells against oxidative/nitrative stress. Oxidative/nitrative stress observed in schizophrenia may be caused partially by the treatment of patients with various antipsychotics. The aim of our study was to establish the effects of epicatechin and antipsychotics action (the first generation antipsychotic (FGA)--haloperidol and the second generation antipsychotic (SGA)--amisulpride) on peroxidation of plasma lipids in vitro. Lipid peroxidation in human plasma was measured by the level of thiobarbituric acid reactive species (TBARS). The properties of epicatechin were also compared with the action of a well characterized antioxidative commercial polyphenol-resveratrol (3,4',5-trihydroxystilbene) and quercetin (3,5,7,3',4'-pentahydroxyflavone). Amisulpride, contrary to haloperidol (after 1 and 24 h) does not significantly influence the increase of plasma TBARS level in comparison with control samples (P > 0.05). After incubation (1 and 24 h) of plasma with haloperidol in the presence of epicatechin we observed a significantly decreases the level of TBARS (P < 0.001, P < 0.001, respectively). In our other experiments, we found that epicatechin also decreased the amount of TBARS in human plasma treated with amisulpride. In conclusion, the presented results indicate that epicatechin-the major polyphenolic component of green tea reduced significantly human plasma lipid peroxidation caused by haloperidol. Moreover, epicatechin was found to be a more effective antioxidant, than the solution of pure resveratrol or quercetin.

Comprehensive polyoxypregnane glycosides report in Caralluma quadrangula using UPLC-ESI-Q-TOF and their antioxidant effects in human plasma.

ETHNOPHARMACOLOGICAL RELEVANCE: Caralluma quadrangula (Forssk.) N.E.Br. (Syns: = Stapelia quadrangula Forssk. = Monolluma quadrangula Forssk.) is an indigenous member of the genus Caralluma and it is a rather common species on rocky hillsides in the southwestern part of Saudi Arabia. Several members of this genus have found medicinal uses in the treatment of rheumatism, diabetes, leprosy and as antiseptics and disinfectants. All parts are edible but rather more bitter and can cause diarrhea. AIM OF THE STUDY: The present report was tentatively elucidated the structure of acylated and non-acylated polyoxypregnane glycosides from Caralluma quadrangula. MATERIALS AND METHODS: The analyses were performed using an electrospray-ionization quadrupole time-of-flight (ESI-Q-TOF) mass spectrometer in both positive and negative ionization modes to explore fragmentation pathways. The antioxidant and prooxidant properties of the different mobility portions of human plasma were evaluated in vitro using thiobarbituric acid reactive substance assay (TBARS). RESULTS: The analyses showed sixty-five characteristic ion peaks which could be more efficient to assignment the aglycones and fragmentation sequences of sugar moieties. The used ionization modes provided consistent and/or complementary information for most of the pregnane glycosides, their fragmentation sequences, and their aglycones. A DFT Study was performed to elucidate the neutral loss of H2O molecules sequences from aglycones and the esterification linkage. CONCLUSIONS: This report could be useful to reduce material consuming and time in phytochemistry analysis of the different medicinal plants. The two portions significantly depleted TBARS were subjected to autoperoxidation assay in the presence of hydrogen peroxide.

Quetiapine, olanzapine and haloperidol affect human plasma lipid peroxidation in vitro.

OBJECTIVE: Oxidative injury in schizophrenia may be caused not only by pathophysiological processes but partly also by treatment with antipsychotics. The purpose of the present study was to examine and to compare the effects of quetiapine (QUE), olanzapine (OLA) and haloperidol (HAL), at final concentrations corresponding to doses used for treatment of acute episodes of schizophrenia, on plasma lipid peroxidation in vitro, measured by the level of thiobarbituric acid-reactive substances (TBARS). METHODS: Blood from 30 healthy volunteers was collected into ACD (citric acid/citrate/dextrose) solution. The drugs in form of active substances were dissolved in 0.01% dimethyl sulfoxide, added to plasma at the final concentrations [QUE (175 and 275 ng/ml), OLA (20 and 40 ng/ml), HAL (4 and 20 ng/ml)] and incubated for 1 and 24 h at 37 °C. The level of TBARS was measured spectrophotometrically (according to the Rice-Evans method, 1991). RESULTS: The comparative study in vitro showed that QUE causes a decrease in the TBARS level in plasma, whereas HAL increases the plasma TBARS level. After 24 h of incubation of plasma with QUE or HAL (at lower and higher concentrations),thedifferences in TBARS levels between the drugs were significant (p = 5.9 × 10⁻4, p = 2.2 × 10⁻5, respectively). CONCLUSION: QUE and OLA, contrary to the prooxidative action of HAL, did not induce oxidative stress; moreover, QUE has antioxidant properties.

Evaluation of cytotoxicity of new trans-palladium(II) complex in human cells in vitro.

Studies of cytotoxicity allow to elucidate the mechanisms by which chemical compounds influence cells and tissues. On the basis of the structural analogy between platinum(II) and palladium(II) complexes, a variety of studies on palladium(II) compounds as potential anticancer drugs have been carried out (1, 2). The cytotoxicity was evaluated by MTT assay. Abilities of trans-palladium(II) complex containing diethyl (pyridin-2-ylmethyl)phosphates as non-leaving ligands (trans-[PdCl2(2-pmOpe 2)]) to induce apoptosis and necrosis in normal lymphocytes, A549 cells and HT29 cell lines were performed by use of fluorochrome staining. The obtained results revealed, that the new trans-palladium(II) complex was more cytotoxic against A549 and HT29 tumor cells than on the normal lymphocytes in vitro. The novel complex induces apoptosis in all tested cells, but in lymphocytes to a lesser degree. The compound tested also induced significant amounts of necrotic cells, which exceeded the level of apoptotic cell fractions. The results demonstrate that the trans-Pd(II) complex showed substantial cytotoxic activity against A549 and HT29 tumor cells and indicate that the new trans-palladium(II) complex effectively inhibited cancer cells growth.

Lipid peroxidation in patients with schizophrenia.

AIMS: There is evidence that dysregulation of free radicals metabolism associated with abnormal activities of antioxidative enzymes in schizophrenia can lead to lipid peroxidation in plasma, erythrocytes, blood platelets and cerebrospinal fluid. Injury to neurons in schizophrenia may affect their function, i.e. membrane transport, impairment of energy production in mitochondria, changes in membrane phospholipid composition, alteration of receptors and transporters as well as neurotransmission. The purpose of the present study was to assess the total antioxidant capacity (TAC) and lipid peroxidation (expressed as the level of thiobarbituric acid reactive substances [TBARS]) in plasma from schizophrenic patients taking olanzapine or risperidone. The level of TBARS estimated according to the Rice-Evans method and TAC ([ABTS; 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization assay]) in plasma from schizophrenic patients (DSM-IV criteria for schizophrenia, n = 30, age 18-36) taking olanzapine or risperidone and from healthy volunteers (n = 30) were measured. METHODS: The level of TBARS in plasma from healthy volunteers after incubation with olanzapine or risperidone was also estimated. RESULTS: Significantly lower plasma TAC (P < 0.05) and significantly increased level of TBARS (P < 0.001) in schizophrenic patients were observed. The in vitro study showed that after olanzapine or risperidone (at final concentrations corresponding to doses used in acute episodes of schizophrenia treatment) no changes of plasma lipid peroxidation were found (P > 0.05). The obtained results indicate that the pro-oxidant disturbances occur in schizophrenic patients (acute episode) taking stable doses of olanzapine or risperidone. CONCLUSION: It seems that second-generation antipsychotics (olanzapine and risperidone) are not responsible for increase of plasma lipid peroxidation.

The Effect of Hydrogen Sulfide on Different Parameters of Human Plasma in the Presence or Absence of Exogenous Reactive Oxygen Species.

The main aim of the study is to examine the effect of sodium hydrosulfide (NaHS), an H2S donor, on the oxidative stress in human plasma in vitro. It also examined the effects of very high concentrations of exogenous hydrogen sulfide on the hemostatic parameters (coagulation and fibrinolytic activity) of human plasma. Plasma was incubated for 5-30 min with different concentrations of NaHS from 0.01 to 10 mM. Following this, lipid peroxidation was measured as a thiobarbituric acid reactive substance (TBARS) concentration and the oxidation of amino acid residues in proteins was measured by determining the amounts of thiol groups and carbonyl groups. Hydrogen peroxide (H2O2) and the hydroxyl radical generating oxidation system (Fe/H2O2) were used as oxidative stress inducers. Hemostatic factors, such as the maximum velocity of clot formation, fibrin lysis half-time, the activated partial thromboplastin time (APTT), thrombin time (TT), and international normalized ratio (INR), were estimated. Changes in lipid peroxidation, carbonyl group formation, and thiol group oxidation were detected at high concentrations of H2S (0.1-10 mM), and these results indicate that NaHS (as the precursor of H2S) may have pro-oxidative effects in human plasma in vitro. Moreover, considering the data presented in this study, we suggest that the oxidative stress stimulated by NaHS (at high concentrations: 1-10 mM) is not involved in changes of the hemostatic activity of plasma.

Oxidative Stress and Hemostatic Parameters in Patients With Nephrolithiasis Before and After Ureteroscopic Lithotripsy.

PURPOSE: In patients with nephrolithiasis, oxidative stress, especially lipid peroxidation is observed. Moreover, various invasive methods [including extracorporeal shock wave lithotripsy (ESWL)] for treatment of nephrolithiasis may induce not only the oxidative stress, but they may modulate hemostasis. The study was aimed to evaluate the oxidative damages of lipids and proteins in patients with nephrolithiasis (before and after ureteroscopic lithotripsy - URSL). The aim of the present study was also determine selected parameters of hemostasis in these patients. METHODS: 56 patients with nephrolithiasis and 49 healthy participants were included: 30 men and 26 women (for patient group); 27 men and 22 women (for healthy group). We measured the level of selected typical two biomarkers of oxidative modification of lipids [such as the production of thiobarbituric acid reactive substances (TBARS) and isoprostane concentration (8-isoPGF2α)] and two biomarkers of oxidative damages of proteins (carbonylation and the level of thiol groups) in patients with nephrolithiasis (before and after URSL). The following parameters of hemostasis were measured: blood platelet count, the level of fibrinogen and D-dimer, and coagulation times (the activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) of plasma). RESULTS: Different levels of plasma lipid peroxidation were observed in patients with nephrolithiasis before URSL and after URSL. However, no such difference in the level of oxidative damage to plasma proteins was observed. In addition, the tested hemostasis parameters were not influenced by the presence of nephrolithiasis, nor by treatment with URSL. CONCLUSION: We suggest URSL does not induce the oxidative modifications of plasma proteins and does not change hemostatic parameters in patients with nephrolithiasis.

Evaluation of antioxidant activity of extracts from the roots and shoots of Scutellaria alpina L. and S. altissima L. in selected blood cells.

BACKGROUND: It is widely known that reactive oxygen species (ROS) can cause oxidative damage in cells and have been linked to the pathogenesis of oxidative diseases, such as atherosclerosis, ischemia, neurodegenerative disease, diabetes, or cancer. Recently, much attention has been focused on preventive strategies for oxidative stress and related diseases. Plants represent a source of bioactive compounds whose antioxidant activity may be useful in protecting against pro-oxidative reactions. OBJECTIVES: The study determines the in vitro biological activity of the ethanolic extracts from the shoots and roots of Scutellaria species (S. altissima and S. alpina) in selected blood cells (blood platelets and lymphocytes). MATERIAL AND METHODS: Platelet activity, both resting and after thrombin stimulation, was used to indicate the ability of the plant extracts to inhibit the production of superoxide anion radicals (O2 •-) and platelet lipid peroxidation. The generation of superoxide anion radicals was measured by cytochrome c reduction. Lipid peroxidation in blood platelets was measured by the level of thiobarbituric acid reactive substances (TBARS). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay was used to determine the protective effect of Scutellaria extracts on lymphocyte cells against oxidative damage induced by hydroxyl radicals. RESULTS: Extracts (5-50 µg/mL) containing phenolic compounds from both Scutellaria species distinctly reduced nonenzymatic lipid peroxidation and arachidonic acid metabolism by blood platelets in vitro. When given at the tested concentration, the extracts reduced the generation of O2 •- in resting blood platelets and platelets activated by thrombin in vitro. All Scutellaria extracts (10 µg/mL) containing phenolic compounds also protected human lymphocytes against oxidative stress induced by hydrogen peroxide (H2O2). CONCLUSIONS: The present study suggests that the natural extracts from S. altissima and S. alpina have antioxidant properties and, therefore, may be beneficial in the prevention of diseases in which blood platelets and lymphocytes are involved, i.e., cancer or inflammatory and infective diseases.

Isorhamnetin and its new derivatives isolated from sea buckthorn berries prevent H2O2/Fe - Induced oxidative stress and changes in hemostasis.

The objective of this study is to investigate the biological effects of phenolic compounds extracted from the sea buckthorn berries on oxidative stress and hemostasis. The sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) berries are rich in flavonoids and non-polar compounds. In this study, the activity of the phenolic fraction from the sea buckthorn berries was evaluated in vitro in comparison with three phenolic compounds: isorhamnetin (compound 1) and its two new derivatives: compound 2 (isorhamnetin 3-O-beta-glucoside-7-O-alfa-rhamnoside) and compound 3 (isorhamnetin 3-O-beta-glucoside-7-O-alfa-(3"'-isovaleryl)-rhamnoside). The impact of these phenolic compounds and the phenolic fraction against the effect of the donor of hydroxyl radicals - H2O2/Fe on proteins and lipids in human plasma was measured. Additionally, the aim of the study was to determine the effect of these phenolic compounds and the phenolic fraction on various typical hemostasis parameters. Our results show that the used derivatives of isorhamnetin possess different biological properties (e.g. antioxidant, anti-platelet and anticoagulant). The tested compounds can be seen as new natural beneficial compounds to be used in prevention and treatment of cardiovascular diseases.

The lipid peroxidation in patients with nephrolithiasis before and after extracorporeal shock wave lithotripsy.

AIM: To evaluate the level of lipid peroxidation in patients with nephrolithiasis before and after extracorporeal shock wave lithotripsy (ESWL). MATERIALS & METHODS: Isoprostane concentration (8-isoPGF2α) was measured in urine, and thiobarbituric acid reactive substance production in serum and erythrocytes. In addition, the concentrations of selected compounds (uric acid, glucose and creatinine) were measured in serum. RESULTS: The patients (before and after ESWL) demonstrated significantly higher levels of two different biomarkers of lipid peroxidation compared with the control group. A correlation was identified between increased amounts of uric acid and biomarkers of lipid peroxidation in patients with nephrolithiasis, both before and after ESWL. CONCLUSION: Uric acid may be associated with lipid peroxidation in patients with nephrolithiasis.