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(1) The cytotoxicity and antioxidant activity of different fractions as well as the pro-apoptotic activity of saponin fractions from Eryngium planum L. in SKOV-3 was investigated. (2) In screening studies, the cytotoxicity of six fractions on SKOV-3 was examined by LDH and SRB assays. The most active fractions—triterpenoid saponins—were selected for further investigation. To determine the mechanism of saponin fractions’ cytotoxicity, their ability to induce apoptosis was examined via Annexin V assay. The effect of the saponin fractions on caspase 3 activity was measured using a Caspase 3 Assay Kit. The expression of 84 apoptosis-related genes was investigated in cancer cells exposed to saponin fractions from the roots. The radical scavenging capacity of different fractions was determined via DPPH assay. (3) The pronounced cytotoxic effects in SKOV-3 were demonstrated by saponin fractions from the leaves and roots. Those saponin fractions were chosen for further investigation. The treatment of cancer cell lines with saponins obtained from the roots provoked a significant increase in apoptotic cells. In the SKOV-3 cells, saponins caused upregulation of pro-apoptotic genes and a decrease in anti-apoptotic genes. The activation of caspase 3 was correlated with an increased DFFA expression level in the treated SKOV-3 cells. The most active fractions were phenolic acids from the shoots and roots. (4) To the best of our knowledge, the current study is the first to demonstrate that the barrigenol-type triterpenoid saponin fraction from the roots of E. planum inhibits SKOV-3 cell proliferation and induces apoptosis, which may be regulated by the expression of genes mostly specific to a mitochondria-related pathway.

The present study has tentatively elucidated the structure of two acylated polyoxypregnane glycosides from Caralluma quadrangula (Forssk.) N.E.Br. (CQ). The analyses were performed using an electrospray-ionization quadrupole time-of-flight (ESI-Q-TOF) mass spectrometer in positive ionization modes to explore fragmentation pathways. The used ionization mode provided consistent and/or complementary information for most of the pregnane glycosides, their fragmentation sequences, and their aglycones. Presumably, this is due to higher efficiency, sensitivity, and better selectivity of the mass spectrometry-based method. The present experimental and theoretical report deals with the characteristic fragmentation behaviors of two acylated polyoxypregnane glycosides CQ1 and CQ2 from the aerial parts of C. quadrangular. A DFT Study was performed to elucidate the position of ikemoyl, and benzoyl residues in compounds CQ1 and CQ2, respectively.

The frequent coexistence of obesity and metabolic syndrome in patients with alopecia areata may indicate the common pathogenetic pathway in these conditions with an important role of adipokines. The aim of the study was to evaluate the serum level of adiponectin, resistin and leptin in patients with alopecia areata in comparison to healthy controls. The study included 65 patients with alopecia areata and 71 healthy controls. The concentration of adipokines was determined with the enzyme-linked immunosorbent assay. The mean concentrations of adiponectin and resistin were significantly lower in the sera of patients with alopecia areata when compared to healthy controls (7966 $$\pm$$ ± 4087 vs 9947 $$\pm$$ ± 5692 ng/ml; p = 0.0312 and 11.04 $$\pm$$ ± 3.88 vs 14.11 $$\pm$$ ± 8.69 ng/ml; p = 0.0176, respectively). A negative correlation between the serum level of adiponectin and severity of alopecia tool (SALT) score was observed (r = − 0.26; p $$\pm$$ ± 2499 vs 8525 $$\pm$$ ± 4085 ng/ml; p = 0.0135). No significant difference in the serum concentration of leptin was observed between patients with alopecia areata and healthy controls. The negative correlation between the serum level of adiponectin and hair loss severity indicates that adiponectin may be considered a marker of hair loss severity in alopecia areata. Further studies are needed to evaluate the role of resistin in patients with alopecia areata and its decreased level irregardless of severity or activity of the disease.

Paulownia is a fast-growing tree that produces a huge mass of leaves as waste that can be used as a feed source for ruminants. The previous study showed that phenolic compounds were the most active biological substances in

Adiponectin, resistin and leptin belong to adipokines, a group of molecules secreted mainly by the adipose tissue, which impaired expression may be a missing link between various manifestations of systemic sclerosis. Adiponectin, which is also released in small amounts by the endothelium, possesses anti-inflammatory, anti-fibrotic and protective against endothelial injury properties. Both leptin and resistin exhibit features which are contradictory to adiponectin, as they trigger inflammation and the activation of skin fibroblasts. Epoprostenol is a prostaglandin analogue with powerful vasodilator activity and inhibitory effect on platelet aggregation. The aim of the study was to evaluate whether epoprostenol may have an effect on serum adipokine levels in patients with systemic sclerosis. A total of 27 patients were included in the study and received epoprostenol intravenously (25 µg of per day for 3 consecutive days). Serum concentrations of total adiponectin, resistin and leptin were assessed with enzyme-linked immunosorbent essay (R&D Systems, Minneapolis, MN, USA). In all SSc patients, the basal level of adiponectin was significantly lower compared to healthy controls (mean 6.00 $$\pm $$ 2.81 μg/ml vs. 8.8 $$\pm $$ 4.3 μg/ml, p = 0.02) and basal level of resistin (mean 11.12 $$\pm $$ 3.36 ng/ml vs. 8.54 $$\pm $$ 3.07 ng/ml p = 0.02) was significantly higher than in the control group. The serum concentration of adiponectin increased significantly after treatment with epoprostenol (6.00 $$\pm $$ 2.81 μg/ml vs 9.29 $$\pm $$ 6.05 μg/ml; P = 0.002). The level of resistin and leptin remained unchanged. Epoprostenol infusions up-regulate the serum concentration of adiponectin in patients with systemic sclerosis. In our opinion, future studies on treatments in systemic sclerosis should address the issue of their effect on adipokine metabolism.

BACKGROUND: Ten varieties of alfalfa (Medicago sativa L.) were evaluated for saponin content. Two of the most promising varieties were chosen so that their effect on rumen fermentation and methane production could be studied. Initially, four Hohenheim gas tests (HGT) were performed to test the effect of increased levels of total saponin extracted from the two alfalfa cultivars (Kometa and Verko) – either as fresh material or ensiled – on the total bacteria, total protozoa, methane emission, and selected methanogenic population. Afterwards, seven particular saponins were extracted from fresh alfalfa of the Kometa variety and tested in 24 h batch fermentation culture experiments. RESULTS: The ensiled forms of both the Verko and Kometa alfalfa varieties seem to be good sources of saponin, capable of reducing methane production (P

Introduction Systemic sclerosis (SSc) is an autoimmune connective tissue disease with distinguished fibrosis of the skin and internal organs. Vascular damage, immune dysregulation and fibroblasts activation contribute to SSc pathogenesis. Peroxisome proliferator-activated receptor γ (PPAR-γ) can be a link between cell metabolism and fibrosis in SSc due to its anti-fibrotic and immunomodulatory properties. Aim To measure the serum level of PPAR-γ in SSc patients and correlate it with the SSc subtype, hs-CRP, disease duration, vascular and internal organ involvement. Material and methods Twenty-two SSc patients (15 limited SSc, 7 diffuse SSc) matched with healthy controls were analysed. Clinical and laboratory data were collected including specific antibodies, interstitial lung disease, oesophageal involvement, digital pitting scars, disease duration, Raynaud's phenomenon (RP) and modified Rodnan skin score (mRSS). PPAR-γ levels were analysed by ELISA. Statistical analysis was performed with χ2, Student's t-test and Mann-Whitney-U test. Pearson and Spearman correlation analyses were used to establish variables association. The significance threshold was set at p Results PPAR-γ concentration was elevated in SSc patients in comparison to controls (p = 0.007) with the highest difference for diffuseSSc (p = 0.004) with significantly elevated mRSS. No association between PPAR-γ levels and hs-CRP, internal organ and vascular involvement, disease duration, autoantibodies and RP onset was found. Conclusions The present study revealed elevated serum PPAR-γ in SSc patients, in particular those with a diffuse form, presenting highest mRSS and lowest BMI. Whether circulating PPAR-γ originates from atrophic adipose tissue, reperfused vessels or ischemic tissues needs assessing. Also the biological meaning or effect of elevated serum PPAR-γ requires further studies.

Uncontrolled blood platelet activation is an important risk factor of cardiovascular disease (CVDs). Various studies on phenolic compounds indicate that they have a protective effect on the cardiovascular system through different mechanisms, including the reduction of blood platelet activation. One of the plants that is particularly rich in phenolic compounds is sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson). The aim of the present study in vitro was to determine the anti-platelet properties of crude extracts isolated from leaves and twigs of E. rhamnoides (L.) A. Nelson in whole blood using flow cytometric and total thrombus-formation analysis system (T-TAS). The aim of our study was also analyze of blood platelet proteoms in the presence of different sea buckthorn extracts. A significant new finding is a decrease surface expression of P-selectin on blood platelets stimulated by 10 µM ADP and 10 µg/mL collagen, and a decrease surface expression of GPIIb/IIIa active complex on non-activated platelets and platelets stimulated by 10 µM ADP and 10 µg/mL collagen in the presence of sea buckthorn leaf extract (especially at the concentration 50 µg/mL). The twig extract also displayed antiplatelet potential. However, this activity was higher in the leaf extract than in the twig extract in whole blood. In addition, our present findings clearly demonstrate that investigated plant extracts have anticoagulant properties (measured by T-TAS). Therefore, the two tested extracts may be promising candidates for the natural anti-platelet and anticoagulant supplements.

Blood platelets play a crucial role in hemostasis, the process responsible for keeping blood flowing in the circulatory system. However, unnecessary platelet activation can lead to aggregation at the site of atherosclerotic plaque rapture and the formation of a thrombus, which promotes atherothrombotic diseases. Various dietary components, such as phenolic compounds, are known to demonstrate antiplatelet and anticoagulant properties, and it is possible that these could form an important element in the prophylaxis and therapy of cardiovascular diseases. Our present study examines the biological activity of isorhamnetin (compound 1) and two isorhamnetin derivatives, compound 2 (3-O-beta-glucoside-7-O-alpha-rhamnoside) and compound 3 (3-O-beta-glucoside-7-O-alpha-(3”’-isovaleryl)-rhamnoside), isolated from the phenolic fraction of sea buckthorn fruit, against human washed blood platelets and human whole blood in vitro. The anti-platelet and anticoagulant potential was determined using (A) flow cytometry, (B) the thrombus-formation analysis system (T-TAS) and (C) colorimetry. The tested flavonoids demonstrated anticoagulant and anti-platelet potential, including anti-adhesive activity, with these effects being more intense in compound 2 than isorhamnetin. Compound 2 inhibited GPIIb/IIIa and P-selectin expression on blood platelets from whole blood, and demonstrated anti-adhesion properties in washed blood platelets and anti-coagulant potential in whole blood, measured by T-TAS.

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

Increased blood platelet activation plays an important role in cardiovascular diseases (CVDs). Recent experiments indicate that certain fruits and vegetables, including onion, garlic, and beetroot, have anti-platelet potential and therefore may reduce the likelihood of CVDs. While vegetables from the Cucuritaceae family are known to exerting beneficial antioxidant and anti-inflammatory effects, their effects on blood platelet activation are poorly understood. Therefore, the aim of the present study was to determine the effect on platelet adhesion of preparations from selected cucurbits: pumpkin (Cucirbita pepo; fruit without seeds), zucchini (Cucurbita pepo convar. giromontina; fruit with seeds), cucumber (Cucumis sativus; fruit with seeds), white pattypan squash (Cucurbita pepo var. patisoniana; fruit without seeds) and yellow pattypan squash (Cucurbita pepo var. patisoniana, fruit without seeds). It also evaluates the activity of these preparations on enzymatic lipid peroxidation in thrombin-activated washed blood platelets by TBARS assay. The study also determines the anti-platelet and anticoagulant properties of these five cucurbit preparations in whole blood by flow cytometry and with the total thrombus-formation analysis system (T-TAS) and evaluates the cytotoxicity of the tested preparations against platelets based on LDH activity. The results indicate that the yellow Cucurbita pepo var. patisoniana preparation demonstrated stronger anti-platelet properties than the other tested preparations, reducing the adhesion of thrombin-activated platelets to collagen/fibrinogen, and inhibiting arachidonic acid metabolism and GPIIb/IIIa expression on 10 µM ADP-activated platelets. None of the preparations was found to cause platelet lysis. Our findings provide new information on the anti-platelet activity of the tested cucurbit preparations and their potential for treating CVDs associated with platelet hyperactivity.

The present study has tentatively elucidated the structure of two acylated polyoxypregnane glycosides from

Bioactive compounds isolated from plants are considered to be attractive candidates for cancer therapy. In this study, we examined the effect of kaempferol, its derivatives, the polyphenol fraction (PF) and an extract (EX) isolated from the aerial parts of Lens culinaris Medik. on DNA damage induced by etoposide in human cells. We also studied the effect of these compounds and their combinations on cell viability. The studies were conducted on HL-60 cells and human peripheral blood mononuclear cells (PBMCs). We used the comet assay in the alkaline version to evaluate DNA damage. To examine cell viability we applied the trypan blue exclusion assay. We demonstrated that kaempferol glycoside derivatives isolated from the aerial parts of Lens culinaris Medik. reduce DNA damage induced by etoposide in PBMCs, but do not have an impact on DNA damage in HL-60 cells. We also showed that kaempferol induces DNA damage in HL-60 cells and leads to an increase of DNA damage provoked by etoposide. Our data suggest that kaempferol derivatives can be further explored as a potential agent protecting normal cells against DNA damage induced by etoposide. Moreover, kaempferol's ability to induce DNA damage in cancer cells and to increase DNA damage caused by etoposide may be useful in designing and improving anticancer therapies.

Systemic sclerosis (SSc) is a chronic connective tissue disease characterized by progressive fibrosis, vascular impairment and immune abnormalities. In recent years, adipokines (mediators synthetized by adipose tissue) have been indicated as a possible missing link in the pathogenesis of SSc. The aim of this study was to investigate the serum concentration of metabolic adipose tissue factors: adiponectin, resistin, leptin and endothelial proteins: endothelin-1, fractalkine and galectin-3 in patients with systemic sclerosis. The study included 100 patients with confirmed SSc diagnosis and 20 healthy individuals. The concentration of respective proteins was determined by enzyme-linked immunosorbent assay. The following markers showed statistically significant increased mean concentrations in patients with SSc in comparison to healthy control: resistin (13.41 vs 8.54 ng/mL; P = 0.0012), endothelin-1 (1.99 vs 1.31 pg/mL; P = 0.0072) and fractalkine (2.93 vs 1.68 ng/mL; P = 0.0007). Elevated serum levels of galectin-3 (4.54 vs 3.26 ng/mL; P = 0.0672) and leptin (19,542 vs 14,210 pg/mL; P = 0.1817) were observed. Decreased concentration of adiponectin was found in patients with SSc (5150 vs 8847 pg/mL; P = 0.0001). Fractalkine and galectin-3 levels were significantly higher in diffuse cutaneous SSc than limited cutaneous SSc subset (3.93 ng/mL vs 2.58 ng/mL, P = 0.0018; 6.86 ng/mL vs 3.78 ng/mL, P = 0.0008, respectively) and correlated positively with modified Rodnan Skin Score in total SSc patients (r = 0.376, P = 0.0009; r = 0.236, P = 0.018, respectively). In conclusion, an increased serum level of resistin associated with increased endothelin-1 and fractalkine level and decreased adiponectin level may indicate a significant role of the adipose tissue in the development and progression of vascular abnormalities in patients with systemic sclerosis. Fractalkine and galectin-3 may participate in promoting and exacerbating the fibrotic process in SSc.

The effect of dandelion root and its various ingredients on the biological functions of blood platelets has not been well investigated. Thus, the objective of our in vitro study was to examine the five root fractions (A–E), differing in chemical composition, for changes induced in the human platelets’ model using selected hemostatic parameters. As a result, the best anti-platelet potential was shown by preparation enriched with hydroxyphenylacaetate inositol esters – PIEs (fraction C). On the other hand, the best overall protective effect against oxidation of lipids and proteins of platelets, and the best inhibitory effect on the generation of O2− were observed in the presence of fraction A (SL-amino acid adducts enriched fraction). Thus, PIEs and SLs rich plant organs, such as dandelion root, may be considered as potentially suitable additives for functional food products used in prophylaxis and treatment of cardiovascular diseases associated with hyperactivation of blood platelets.

Three previously undescribed cardenolides, acovenosigenin A 3-O-α-L-acofriopyranoside (1), 14-anhydroacovenosigenin A 3-O-[β-D-glucopyranosyl-(1″→4′)-O-α-L-acofriopyranoside] (2), and 14-anhydroacovenosigenin A 3-O-[β-D-glucopyranosyl-(1″→4′)-O-α-L-acovenopyranoside] (3), together with the two already known ones, 14-anhydrodigitoxigenin 3-O-β-D-glucopyranoside (4) and acospectoside A (5), were isolated from the leaves of Acokanthera oblongifolia. The influence of cardenolides 1 – 3 and acovenoside A (found in the Acokanthera genus) on three cancer cell lines (HT29, HCT116, and AGS) was also investigated. The most promising results, in comparison with oxaliplatin, were obtained for compound 1, which was found to be highly cytotoxic for all tested cell lines, HT29 (IC50 = 63.49 nM), HCT116 (IC50 = 67.35 nM), and AGS (IC50 = 80.92 nM). Unfortunately, 1 also showed similar toxicity towards normal lymphocytes (IC50 = 98.03 nM).

Triterpene saponins from the leaves of red beet (Beta vulgaris L.) were qualitatively profiled using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Results indicated that red beet leaves contain 11 saponins, including oleanolic acid aglycone and a varying number of sugars plus acetyl or methyl groups. We identified and studied two additional saponins other than the nine reported previously from red beets. Relative quantification was undertaken of saponins in leaves, compared to those in the roots of four cultivars (Egyptian, Forono, Red Sphere, and Round Dark Red). Saponins were quantitated using ultra-performance reverse-phase liquid chromatography (UPLC) coupled with ESI-MS/MS. Leaves of Egyptian and Round Dark Red contained higher total amount of saponins than the roots, whereas relatively higher levels of saponins were observed in roots in cultivars Red Sphere and Forono. The differential accumulation of specific triterpene saponins is indicative of spatially differentiated biosynthesis and/or biological function.

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.

Summary Introduction: Callus and cell suspension cultures are widely applied in investigation of production of high-value secondary metabolites, which may be used as cosmeceuticals, nutraceuticals and pharmaceuticals. Plant cell cultures are promising alternative to intact plant sources for the production of plant-derived drugs of industrial importance. Objective: The aim of the study was to (i) initiate the cell suspension culture of Chaenomeles japonica from homogenous and uniform callus, (ii) stabilize the selected line and (iii) verify its ability to produce the desired groups of secondary metabolites – pentacyclic triterpenoids and polyphenols. Methods: To establish a cell suspension culture, stabilized and homogeneous callus was selected. Cell cultures were systematically passaged every 2 weeks to fresh liquid medium with the same composition. Biomass from cultures at the growth phase and stationary phase was designated for phytochemical research. UHPLC-DAD-MS analyzes were performed. At the same time, their macroscopic and microscopic observations were carried out. Results: Cells of suspension culture line A2 were characterized by the intense divisions. Cell culture extracts (both from the growth phase and stationary phase) contained pentacyclic triterpenoids. In addition, phe-nolic compounds (chlorogenic acid and proanthocyanidins type B) and in a small amount also epicatechin are present in the extract of the cells harvested from the growth phase. In the present studies, three pentacyclic triterpenoids were detected and quantified in the extracts of cell suspensions and callus line A2. Ursolic and oleanolic acids were the main triterpenoids in the studied extracts. The cell suspension culture from the growth phase exhibited the highest content of ursolic, oleanolic, and betulinic acid (separately and together). Conclusion: The cell suspension culture of Chaenomeles japonica is a promising source of pentacyclic triterpenoids.

Laboratory analyses using the 1st generation bioassay were conducted in the years 2013-2014 to investigate the allelopathic potential of wateralcoholic and aqueous extracts from dry weight of rhizomes and roots as well as stems and leaves of Solidago gigantea. Analysed acceptors were two weed species, i.e. monocotyledonous Echinochloa crus-galli and dicotyledonous Amaranthus retroflexus. When the acceptors (E. crus-galli and A. retroflexus) reached the 2-leaf stage (BBCH 12) they were sprayed with wateralcoholic and aqueous extracts (at concentrations of 12.5%, 10%, 5% and 2.5%) obtained from the donor, i.e. S. gigantea. Results indicate an inhibitory effect of wateralcoholic extracts from aboveground parts (leaves and stems) of S. gigantea in relation to dry weight of E. crus-galli and A. retroflexus. The volume of dry weight reduction in acceptors was dependent on the concentration of extracts produced from the donor plant S. gigantea. Dry weight of E. crus-galli and A. retroflexus was reduced most effectively by two concentrations: 12.5% and 10%. In turn, aqueous extracts from rhizomes and roots of S. gigantea, irrespective of the applied concentration, caused an increase in dry weight of E. crus-galli and A. retroflexus. Only aqueous extracts produced from leaves and stems of S. gigantea, irrespective of their concentration, reduced dry weight in only E. crus-galli., W badaniach laboratoryjnych przeprowadzonych z użyciem biotestu I generacji w latach 2013-2014, porównywano potencjał allelopatyczny wyciągów wodnoalkoholowych oraz wodnych uzyskanych z suchej masy kłączy i korzeni oraz łodyg i liści Solidago gigantea. Akceptorami były dwa gatunki chwastów: jednoliścienny Echinochloa crus-galli oraz dwuliścienny Amaranthus retroflexus. W monecie uzyskania przez akceptory (E. crus-galli i A. retroflexus), fazy 2 liści właściwych wykonano oprysk wyciągiem wodno-alkoholowym oraz wodnym, o stężeniu 12,5%, 10%, 5% i 2,5%, otrzymanym z donora, którym było S. gigantea. Uzyskane wyniki badań wskazują na efekt inhibicyjnego odziaływania wyciągów wodno-alkoholowych sporządzonych z nadziemnych części roślin (liści i łodyg) S. gigantea w odniesieniu do suchej masy E. crus-galli oraz A. retroflexus. Wielkość redukcji suchej masy akceptorów uzależniona była od stężenia wyciągów sporządzonych z rośliny donorowej S. gigantea. Najefektywniej suchą masę E. crus-galli oraz A. retroflexus redukowały dwa stężenia: 12,5% oraz 10%. Natomiast wyciągi wodne sporządzone z kłączy i korzeni S. gigantea, niezależnie od zastosowanego stężenia, powodowały zwiększenie suchej masy E. crus-galli oraz A. retroflexus. Jedynie wyciągi wodne otrzymane z liści i łodyg S. gigantea, niezależnie od stężenia, powodowały redukcję suchej masy E. crus-galli.

Systemic sclerosis is a multiorgan autoimmune disease characterized by vasculopathy and tissue fibrosis of unknown etiology. Recently, adipokines (cell signaling proteins secreted by adipose tissue) have attracted much attention as a cytokine family contributing to the various pathological processes of systemic sclerosis. Adipokines, such as leptin, adiponectin, resistin, adipsin, visfatin or chemerin are a heterogenic group of molecules. Adiponectin exhibits anti-fibrotic features and affects inflammatory reactions. Leptin promotes fibrosis and inflammation. Resistin was linked to vascular involvement in systemic sclerosis. Visfatin was associated with regression of skin lesions in late-stage systemic sclerosis. Chemerin appears as a marker of increased risk of impaired renal function and development of skin sclerosis in the early stage of systemic sclerosis. Vaspin was indicated to have a protective role in digital ulcers development. Novel adipokines-adipsin, apelin, omentin and CTRP-3-are emerging as molecules potentially involved in SSc pathogenesis. Serum adipokine levels may be used as predictive and diagnostic factors in systemic sclerosis. However, further investigations are required to establish firm correlations between distinct adipokines and systemic sclerosis.

Strawberry (SB), black currant (BC), and raspberry seed (RB) residues were used in 3 experiments to study their effects on ruminal fermentation, methane concentration, and fatty acid (FA) proportions in the ruminal fluid and milk of dairy cows. Initially, a batch fermentation in vitro study (experiment 1) was performed to investigate the effects of the 3 berry residues on basic ruminal fermentation parameters. Total volatile fatty acid concentrations, including acetate, propionate, and butyrate, increased in the BC group compared with other treatments. Based on the preliminary in vitro results, 2 consecutive in vivo experiments were conducted using 4 Polish Holstein-Friesian cows fitted with rumen cannulas (experiment 2) and 30 lactating Polish Holstein-Friesian dairy cows (experiment 3) in a replicated 2 × 2 crossover design. Cows in both experiments received a partial mixed ration (PMR) in 2 variants: (1) a control diet of PMR + 2 kg of concentrate (control); (2) PMR + 2 kg of BC seed residues (BC). The BC diet did not mitigate ruminal methane production. Ruminal fermentation (experiment 2) was not affected by the BC diet; however, the concentrations of C18:1 trans-11 and C18:2 cis-9,trans-11 increased significantly by 91 and 131%, respectively. Likewise, concentrations of total trans C18:1 and total monounsaturated FA in ruminal fluid were increased significantly by BC seed residues. In experiment 3, BC significantly increased milk fat C18:1 trans-11, C18:2 cis-9,trans-11, n-3, n-6, and polyunsaturated FA concentrations without affecting milk production performance. In conclusion, the amount (2 kg/d) of BC used in this study did not adversely affect ruminal fermentation or milk production and composition. However, using BC increased proportions of unsaturated FA and conjugated linoleic acid in milk. Although dietary BC did not exert a strong methane inhibition effect, it could represent an inexpensive alternative concentrate to improve beneficial FA in milk without negative effects on rumen fermentation and production parameters in dairy cows. Incorporation of berry seed residues in diets would be profitable economically and nutritionally for dairy cattle production.

We investigated the conditions leading to the accumulation of saponins in plants using in vitro cultures of Eryngium planum L. The aim was to establish a system that produced saponins of the same quality as those found in field-grown plants and to enhance their accumulation in the laboratory. We observed that the content of saponins in organs from in vitro plantlets and plants grown in an experimental plot after micropropagation was higher (up to 8.7 times) compared with plants from natural sites. Untreated callus accumulated saponins in amounts similar to those of the roots from field-grown plants. Applying sucrose to the growth medium at higher concentrations (40, 50, or 60 g L−1) resulted in enhanced saponin production in callus (1.2–1.7-fold). The elicitation with 100 µmol L−1 methyl jasmonate of callus grown on Murashige and Skoog medium supplemented with 30 g L−1 sucrose resulted in a 1.2-fold increased saponin accumulation compared with callus maintained on the same medium (30 g L−1 sucrose). This study provides the first evidence that triterpenoid saponins can be biosynthesized in in vitro plantlets and dedifferentiated cultures of E. planum.

Although the major components of various organs of sea buckthorn have been identified (particularly phenolic compounds), biological properties of many of these phytochemicals still remain poorly characterized. In this study, we focused on the chemical composition and biological activity of preparations that were obtained from sea buckthorn twigs and leaves. The objective was to investigate cytotoxicity of these preparations against human fibroblast line HFF-1, using MTT reduction assay, their anti- or pro-oxidant activities against the effects of a biological oxidant -H2O2/Fe—on human plasma lipids and proteins in vitro (using TBARS and carbonyl groups as the markers of oxidative stress). Antimicrobial activity of the tested preparations against Gram-positive (Staphylococcus aureus, S. epidermidis, Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), as well as against fungi (Candida albicans, C. glabrata) by the EUCAST-approved broth microdilution method, followed by growth on solid media, were also assessed. Our analysis showed significant differences in chemical composition and biological properties of the tested preparations (A–F). All tested preparations from sea buckthorn twigs (D–F) and one preparation from sea buckthorn leaves (preparation C) may be a new source of phenolic antioxidants for pharmacological and cosmetic applications.

Sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) is a shrub growing in coastal areas. Its organs contain a range of bioactive substances including vitamins, fatty acids, various micro and macro elements, as well as phenolic compounds. Numerous studies of sea buckthorn have found it to have anticancer, anti-ulcer, hepatoprotective, antibacterial, and antiviral properties. Some studies suggest that it also affects the hemostasis system. The aim of the study was to determine the effect of six polyphenols rich and triterpenic acids rich fractions (A–F), taken from various organs of sea buckthorn, on the activation of blood platelets using whole blood, and to assess the effect of the tested fractions on platelet proteins: fraction A (polyphenols rich fraction from fruits), fraction B (triterpenic acids rich fraction from fruits), fraction C (polyphenols rich fraction from leaves), fraction D (triterpenic acids rich fraction from leaves), fraction E (polyphenols rich fraction from twigs), and fraction F (triterpenic acids rich fraction from twigs). Hemostasis parameters were determined using flow cytometry and T-TAS (Total Thrombus-formation Analysis System). Additionally, electrophoresis was performed under reducing and non-reducing conditions. Although all tested fractions inhibit platelet activation, the greatest anti-platelet activity was demonstrated by fraction A, which was rich in flavonol glycosides. In addition, none of the tested fractions (A–F) caused any changes in the platelet proteome, and their anti-platelet potential is not dependent on the P2Y12 receptor.

Background The field bean (Vicia faba) is a valuable fodder plant of the Fabaceae family, grown as a main crop for its seed yield. Its phytochemical profile is characterized by the presence of a range of compounds with various biological activities. Purpose The present study investigates the phytochemical profile of the extract from mature seeds of Vicia faba var. minor and examines its impact on preventing oxidative damage to various lipids, protein and DNA molecules in vitro. Methods Human plasma was treated with H2O2/Fe (an OH. donor) to induce oxidative damage to lipids and proteins, and the plant extract was then added. As oxidative stress may influence the biological activity of plasma, e.g. coagulation, and influence cardiovascular disease, the study also examined the effect of the plant extract on coagulation and monoamine oxidase activity (MAO, EC 1.4.3.4). Results The tested extract exerted a protective effect on plasma lipids and proteins treated with H2O2/Fe. However, while it appears to effectively protect the DNA in peripheral blood mononuclear cells from oxidative damage, it did not induce changes in the coagulation process, and significantly reduced MAO activity when applied at 1, 5, and 10 µg/mL. It is possible that the observed antioxidant potential may be due to the complex chemical composition of the extract: the phytochemical profile demonstrated a range of phenolic compounds, including catechins. Conclusion Our findings indicate that extract from mature seeds of V. faba var. minor may be a promising source of antioxidants in multiple applications, including diseases associated with oxidative stress; however, more studies based on in vitro and in vivo models are needed to determine its biological properties.

Systemic sclerosis (SSc) is a connective tissue disease characterized by endothelial cell damage, perivascular inflammation and tissue hypoxia. Angiopoietin-like protein 4 (ANGPTL4) has been demonstrated to affect vascular permeability, inflammation and oxidative stress, thus may contribute to SSc pathogenesis.The aim of the study was to evaluate serum ANGPTL4 in systemic sclerosis and correlate it with disease subtype (localized and diffuse, lcSSc and dcSSc respectively), disease duration, skin fibrosis and internal organ involvement.Twenty-two patients with systemic sclerosis (15 lcSSc, 7 dcSSc) and thirteen healthy controls were analyzed. Clinical and laboratory data were collected including modified Rodnan Skin Score (mRSS), Raynaud's phenomenon, disease duration, digital pitting scars, oesophageal involvement and interstitial lung disease. ANGPTL4 sera concentrations were measured by ELISA.Patients with systemic sclerosis had lower ANGPTL4 serum levers in comparison to healthy controls, however without statistical significance (160.15 ± 117.53 vs. 127.15 ± 83.58 ng/ml; p=0.64). No association between ANGPTL4 levels and disease subtype, disease duration, severity of skin involvement (mRSS) and Raynaud's phenomenon onset was found.This is the first study evaluating the serum concentration of ANGPTL4 in patients with systemic sclerosis. This study contributes to still undetermined role of ANGPTL4 in the development or progression of systemic sclerosis. Therefore the role of ANGPTL4 in hypoxia-related diseases such as systemic sclerosis needs further research.

Many studies show that saponins isolated from various plants have a cytotoxic effect on cancer cells inducing apoptosis and autophagy. On the other hand, saponins also exhibit a number of beneficial properties, such as antioxidant properties. Thus, saponins can be considered both in terms of their therapeutic and protective effects during anticancer treatment. In this study, we investigated the effect of the saponin fraction isolated from sea buckthorn (Elaeagnus rhamnoides (L.) A. Nelson) leaves on the viability of HL-60 cancer cells using resazurin assay and its ability to induction of apoptosis with Annexin V-FITC and propidium iodide (PI) double staining. Moreover, we studied its effect on the oxidative stress induced by H2O2, and anti-platelet and anticoagulant potential in whole blood using T-TAS, a microchip-based flow chamber system. We observed that the saponin fraction significantly decreased the viability of HL-60 cells at the concentration above 50 µg/mL and induced apoptosis at the concentration of 100 µg/mL. Moreover, we observed that saponin fraction used at lower concentrations, such as 0.5 and 1 µg/mL, stimulated HL-60 cells and increased their viability. The saponin fraction also decreased the level of free radicals and reduced oxidative DNA damage measured by the comet assay. However, at high concentration of oxidant H2O2 equal 5 mM, we noticed that the saponin fraction at 50 µg/mL increased the level of free radicals in HL-60 cells. We also demonstrated anticoagulant potential of the saponin fraction at the concentration of 50 µg/mL. Our results indicate that the saponin fraction obtained from sea buckthorn leaves can show both chemotherapeutic and chemoprotective potential.

Context Bufadienolide compounds occur in many plants and animal species and have strong cardiac and anti-inflammatory properties. The compounds have been recently investigated for cytotoxic and antitumor activity. Objective The cytotoxic effect of bersaldegenin-1,3,5-orthoacetate – a bufadienolide steroid occuring in plants from Kalanchoe genus (Crassulaceae), was evaluated with cervical cancer HeLa cells in vitro. Materials and methods The cytotoxic activity of the compound (at 0.1–20.0 μg/mL) on the cells was determined by Real-Time Cell Analysis (RTCA) system for 24 h. The estimation of cell cycle arrest, reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential (MMP), and caspases-3/7/9 activity in the HeLa cells treated with the compound was done by flow cytometry and luminometric technique. DNA damage in the cells was estimated by immunofluorescence staining and the comet assay with etoposide as a positive control. Results The compound had strong effect on the cells (IC50 = 0.55 μg/mL) by the suppression of HeLa cells proliferation in G2/M phase of cell cycle and induction of cell death through double-stranded DNA damage and reactive oxygen species overproduction. Furthermore, we did not observe an increase in the activity of caspase-3/7/9 in the treated cells as well as a decrease in cellular mitochondrial membrane potential. Gene expression analysis revealed the overexpression of NF-Kappa-B inhibitors genes (>2-fold higher than control) in the treated cells. Conclusions Bersaldegenin-1,3,5-orthoacetate induces cell cycle arrest and caspase-independent cell death through double-stranded DNA damage. These results are an important step in further studies on cell death signalling pathways induced by bufadienolides.

Lychnis flos-cuculi L., a species with potential medicinal value, contains flavonoids, phenolic acids, triterpenoid saponins and ecdysteroids. In this study, the antioxidant activity of plant material of L. flos-cuculi obtained from in vitro cultures compared to that of intact plants from the natural site has been evaluated for the first time. Phytochemical screening of the in-vitro-derived material by ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) confirmed the presence of the aforementioned metabolite classes. The aqueous methanolic extracts from in-vitro-derived plant material and the organs of intact plants were analyzed using spectrophotometric methods to quantify total phenolics, phenolic acids and flavonoids, and determine the preliminary antioxidant activity by ferric reducing antioxidant potential (FRAP) and DPPH radical scavenging activity assays. The results showed that the inflorescence (Ns-F), and flowering herb of both plants gathered from natural habitat (Ns-H) and in-vitro-derived plants from the experimental plot (ExV-H) are the materials richest in polyphenols (195.4, 113.47, 112.1 mg GAE g−1 d.w., respectively), and demonstrate the highest antioxidant activity (20.14, 11.24, and 11.46 mg AAE g−1 d.w.). The extract from callus exhibited the lowest polyphenol content and antioxidant potential. The contents of total phenolics, flavonoids and phenolic acids correlate with the results of the antioxidant capacity of L. flos-cuculi extracts.

Systemic sclerosis (SSc) is an autoimmune disease characterized by vasculopathy and uncontrolled cutaneous and internal organs fibrosis. Diagnosis of SSc in an early phase can be difficult because of a lack of typical symptoms. The delay in diagnosis and treatment of SSc may lead to uncontrolled progression of the disease, thus identification of possible early indicators of skin and organ involvement to prevent their further damage is necessary. The aim of this study is to review the latest biomarkers of organ involvement in SSc. In patients with lung fibrosis lung-epithelial-derived surfactant protein (SP-D), the glycoprotein Krebs von den Lungen-6 (KL-6), and chemokine ligands 2, 4 and 18 (CCL2, CXCL4, CCL18) are elevated, while in patients with skin fibrosis serum levels of heat shock protein 27 (Hsp27), interleukin 16 (IL-16), and IgG–galactosylation ratio are increased. Adiponectin concentration is inversely correlated with the intensity of cutaneous fibrosis. Skin gene profiling also seems very promising. In patients with heart involvement increased serum levels of brain natriuretic peptide (BNP) are present, as well as raised Midkine and Follistatin-like 3 (FSTL3) proteins, ratios of Cu/Se and ceruloplasmin(CP) /Circulating selenoprotein P(SELENOP) and higher whole blood viscosity level. Elevated calprotectin levels are found in individuals with gastrointestinal involvement. Increased levels of chemerin and ARA autoantibodies are associated with renal involvement, whereas high levels of adhesion molecules are found in patients with scleroderma renal crisis (SRC). Currently there are no biomarkers in use that can specifically identify the early involvement of organs.

Background Paulownia Clone in Vitro 112, also known as Oxytree is a hybrid of Paulownia elongata and Paulownia fortunei, developed under laboratory conditions. Its seeds are sterile, making it a noninvasive variety that can only be propagated in the laboratory. In China, species from the Paulownia genus (Paulowniaceae) are widely used in traditional medicine for the treatment of infectious diseases, such as gonorrhea and erysipelas. It has a broad spectrum of bioactivity, including neuroprotective, antioxidant, antibacterial, antiphlogistic, antiviral, and cytotoxic actions. However, the antiplatelet potential of Paulownia Clone in Vitro 112 has not yet been described. Study design The aim of our study was thus to examine the effect of an extract and four fractions from leaves of Paulownia Clone in Vitro 112 on various parameters of platelet activation in an in vitro model. Methods Composition of the investigated extract and fractions was determined by UHPLC-UV-MS. The following parameters of platelet activation were investigated: nonenzymatic lipid peroxidation in resting platelets; enzymatic lipid peroxidation (AA metabolism) in platelets activated by thrombin; superoxide anion (O2−.) generation in the resting and activated platelets; platelet adhesion to collagen type I and fibrinogen; platelet aggregation stimulated by various physiological agonists, such as ADP, collagen, and thrombin. The effect of the extract and fractions on extracellular LDH activity, a marker of cell damage, was also determined. Results Verbascoside a phenylethnanoid glycoside, was the main secondary metabolite of the extract from leaves of oxytree (constituting approximately 45 % of all compounds). There were also iridoids, such as catalpol, aucubin, and 7-hydroxytomentoside, as well as flavonoids, such as luteolin and apigenin glycosides. Moreover, the extract had stronger antiplatelet properties than the fractions. For example, the extract at 10 μg/mL inhibited five parameters of platelet activation. Conclusions Our results show that Paulownia Clone in Vitro 112 leaves are a new valuable source of compounds with antiplatelet potential.

Dandelion (Taraxacum officinale L.) roots, leaves, and flowers have a long history of use in traditional medicine. Compared to the above organs, dandelion fruits are the least known and used. Hence, the present paper was aimed at the phytochemical analysis of T. officinale fruit extract and estimating its antiradical, antiplatelet, and antioxidant properties related to hemostasis. Methanolic extract of fruits (E1), enriched with polyphenols (188 mg gallic acid equivalents (GAE)/g), was successfully separated into cinnamic acids (E2, 448 mg GAE/g) and flavonoids (E3, 377 mg GAE/g) extracts. Flavonoid extract was further divided into four fractions characterized by individual content: A (luteolin fraction, 880 mg GAE/g), B (philonotisflavone fraction, 516 mg GAE/g), C (flavonolignans fraction, 384 mg GAE/g), and D (flavone aglycones fraction, 632 mg GAE/g). High DPPH radical scavenging activity was evaluated for fractions A and B (A &gt, B &gt, Trolox), medium for extracts (Trolox &gt, E3 &gt, E2 &gt, E1), and low for fractions C and D. No simple correlation between polyphenol content and antiradical activity was observed, indicating a significant influence of qualitative factor, including higher anti-oxidative effect of flavonoids with B-ring catechol system compared to hydroxycinnamic acids. No cytotoxic effect on platelets was observed for any dandelion preparation tested. In experiments on plasma and platelets, using several different parameters (lipid peroxidation, protein carbonylation, oxidation of thiols, and platelet adhesion), the highest antioxidant and antiplatelet potential was demonstrated by three fruit preparations&ndash, hydroxycinnamic acids extract (E2), flavonoid extract (E3), and luteolin fraction (A). The results of this paper provide new information on dandelion metabolites, as well as their biological potential and possible use concerning cardiovascular diseases.

The aim of this study was to provide detailed insight into the chemical composition and activity of five cucurbit vegetable preparations (pumpkin, zucchini, cucumber, white and yellow pattypan squash), each containing various phytochemical compounds with potential use against oxidative stress induced by the hydroxyl radical donors in human plasma in vitro. We studied the antiradical capacity of vegetable preparations using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method. As oxidative stress may induce changes in hemostasis, our aim included the determination of their effect on three selected hemostatic parameters of plasma, which are three coagulation times: PT (prothrombin time), APTT (activated partial thromboplastin time) and TT (thrombin time). However, none of used vegetable preparations changed APTT, PT or TT compared to the control. The phytochemical composition of the tested preparations was determined by UPLC-ESI-QTOF-MS. In our in vitro experiments, while all five tested preparations had antioxidant potential, the preparation from yellow pattypan squash showed the strongest potential. All cucurbit vegetable preparations inhibited lipid peroxidation. Only zucchini did not have an effect on protein carbonylation and only yellow pattypan squash inhibited thiol oxidation. The antioxidant activity of cucurbits appears to have triggered significant interest in multiple applications, including CVDs (cardiovascular diseases) associated with oxidative stress, which can be treated by supplementation based on these vegetables.

Due to the richness of bioactive substances and easy accessibility, sea-buckthorn can be an ingredient of currently popular functional food supporting anti-cancer therapy. Low-polarity fractions from fruit (OL), twigs (GL) and leaves (LL) were investigated. Compared to the previous scientific reports a more detailed analysis of the chemical composition of individual fractions was performed. Cytotoxicity of low-polarity fractions has been investigated and activity compared in human tumor and normal cells cultured in vitro. The genotoxicity and pro-apoptotic properties of low-polarity fractions were also followed on selected cell lines that had proved to be the most sensitive. In the proposed research model being tested, low-polarity fractions act cytotoxically, even 3 times more strongly in cancer cells than normal ones. Measurement of caspase 3/7 activity indicated that cell death occurs through apoptosis. Furthermore, high concentrations of low-polarity fractions have moderate genotoxic properties. Data obtained on the biological properties of low-polarity fractions from sea-buckthorn show that these fractions can potentially support cancer cells elimination. Phytotochemical analysis indicates the key role of the triterpenoids in this process.

Benzoxazinoids (BXs) are specialized metabolites with protective properties that are synthesized predominantly by Poaceae species, including rye (Secale cereale). Among factors known to influence BXs production, prolonged low temperature has not been studied previously. In this study, the influence of cultivation at 4 &deg, C, which is essential for vernalization, on the concentration of BXs (HBOA, DIBOA, GDIBOA, DIMBOA, GDIMBOA, and MBOA) and the expression level of genes involved in the BX biosynthesis pathway (ScBx1&ndash, ScBx5 and ScIgl) in three rye inbred lines was investigated. After cultivation for seven weeks at 4 &deg, C, the expression level of all analyzed genes and BX concentrations had decreased compared with those at the initiation of treatment (21 days after germination) in control and cold-treated plants. At this time point, the decrease in BX concentrations and gene expression was lower in cold-treated plants than in untreated plants. In contrast, at 77 days after germination, the gene expression levels and BX concentrations in untreated plants had generally increased. Investigation of the vernalization impact on rye BXs accumulation, as well as on Bx gene expression, may aid with determination of the most suitable winter lines and cultivars of rye for cultivation and breeding purposes.

Background: The Paulownia Clone in Vitro 112, known as oxytree or oxygen tree, is a hybrid clone of the species Paulownia elongata and Paulownia fortunei (Paulowniaceae). The oxytree is a fast-growing hybrid cultivar that can adapt to wide variations in edaphic and climate conditions. In this work, Paulownia Clone in Vitro 112 leaves were separated into an extract and four fractions (A&ndash, D) differing in chemical content in order to investigate their chemical content using LC-MS analysis. The extract and fractions were also evaluated for their anticoagulant and antioxidant properties in a human plasma in vitro. Results: The Paulownia leaf extract contained mainly phenolic compounds (e.g., verbascoside), small amounts of iridoids (e.g., aucubin or 7-hydroxytometoside) and triterpenoids (e.g., maslinic acid) were also detected. Our results indicate that the extract and fractions have different effects on oxidative stress in human plasma treated with H2O2/Fe in vitro, which could be attributed to differences in their chemical content. For example, the extract and all the fractions, at the two highest concentrations of 10 and 50 &micro, g/mL, significantly inhibited the plasma lipid peroxidation induced by H2O2/Fe. Fractions C and D, at all tested concentrations (1&ndash, 50 &micro, g/mL) were also found to protect plasma proteins against H2O2/Fe-induced carbonylation. The positive effects of fraction C and D were dependent on the dose. Conclusions: The extract and all four fractions, but particularly fractions C and D, which are rich in phenolic compounds, are novel sources of antioxidants, with an inhibitory effect on oxidative stress in human plasma in vitro. Additionally, the antioxidant potential of fraction D may be associated with triterpenoids.