Phytochemical, Pharmacological, and Molecular Docking Study of Dry Extracts of Matricaria discoidea DC. with Analgesic and Soporific Activities.

Pineapple weed (Matricaria discoidea DC.) is a widespread plant in Europe and North America. In ethnomedicine, it is well-known for its anti-inflammatory and spasmolytic activities. The aim of this research was to develop novel methods of M. discoidea processing to obtain essential oil and dry extracts and to investigate their phytochemical compositions. Moreover, the molecular docking of the main substances and the in vivo studies on their soporific and analgesic activities were conducted. The essential oil and two dry extracts from M. discoidea were prepared. A total of 16 phenolic compounds (seven flavonoids, seven hydroxycinnamic acids, and two phenolic acids) in the dry extracts were identified by means of UPLC-MS/MS. In the essential oil, nine main terpenoids were identified by gas chromatography (GC). It was shown that phenolic extraction from the herb was successful when using 70% ethanol in a triple extraction method and at a ratio of 1:14-1:16. The in vivo studies with rodents demonstrated the analgesic activity of the M. discoidea extracts and improvements in the sleep of animals. The dry extracts of M. discoidea did not show any toxicity. The molecular docking analysis showed a high probability of COX-1,2 inhibition and NMDA receptor antagonism by the extracts.

Multi-Omics Analysis Revealed Increased De Novo Synthesis of Serine and Lower Activity of the Methionine Cycle in Breast Cancer Cell Lines.

A pipeline for metabolomics, based on UPLC-ESI-MS, was tested on two malignant breast cancer cell lines of the sub-types ER(+), PR(+), and HER2(3+) (MCF-7 and BCC), and one non-malignant epithelial cancer cell line (MCF-10A). This allowed us to quantify 33 internal metabolites, 10 of which showed a concentration profile associated with malignancy. Whole-transcriptome RNA-seq was also carried out for the three mentioned cell lines. An integrated analysis of metabolomics and transcriptomics was carried out using a genome-scale metabolic model. Metabolomics revealed the depletion of several metabolites that have homocysteine as a precursor, which was consistent with the lower activity of the methionine cycle caused by lower expression of the AHCY gene in cancer cell lines. Increased intracellular serine pools in cancer cell lines appeared to result from the over-expression of PHGDH and PSPH, which are involved in intracellular serine biosynthesis. An increased concentration of pyroglutamic acid in malignant cells was linked to the overexpression of the gene CHAC1.

Evaluation of the Cardiac Electrophysiological and Haemodynamic Effects of Elsholtzia ciliata Essential Oil on Swine.

The demand for the development of novel medicines with few side effects and no proarrhythmic properties is increasing. Extensive research on herbal extracts has been conducted with the expectation that the compounds will exert precise effects without harmful side effects. Elsholtzia ciliata (Thunb.) Hyl. essential oil (EO) possesses antiarrhythmic properties similar to those of class 1B antiarrhythmics, such as prolonging myocardial activation of the QRS complex and shortening the QT interval. In this study, we determined the kinetic profile of EO phytocompounds and the effects of EO on heart electrical activity and arterial blood pressure. For this study, we chose to use local breed pigs that were anaesthetized. The effects of an intravenous bolus of EO on ECG parameters, arterial blood pressure, heart rate variability, and blood levels of haematological and biochemical parameters were registered and evaluated. Following an intravenous injection of a bolus, EO exerted a vasodilatory effect, resulting in significant reductions in arterial blood pressure. EO also increased the heart rate and altered ECG parameters. The bolus of EO prolonged the QRS complex, shortened the QT interval, and nonmonotonically altered the PQ interval. After the administration of a bolus of EO, the activity of the autonomic nervous system was altered. This study confirms that EO possesses similar properties to class 1B antiarrhythmics and exerts a hypotensive effect; it reduces arterial blood pressure possibly by modulating peripheral vascular resistance.

The Phytochemical Profile and Anticancer Activity of Anthemis tinctoria and Angelica sylvestris Used in Estonian Ethnomedicine.

The aerial parts of Anthemis tinctoria L. and Angelica sylvestris L. and the roots of A. sylvestris have been used as traditional anticancer remedies in Estonian ethnomedicine. The aim of this study was to investigate content of essential oils (by gas chromatography) and polyphenolic compounds (using two different methods of high performance liquid chromatography-mass spectrometry (HPLC-MS)) of both plant species, as well as the in vitro anti-cancer effects of their essential oils and methanolic extracts. The average (n = 5 samples) yield of essential oils was 0.15%, 0.13%, and 0.17%, respectively. The principal compounds of the essential oil from the aerial parts of A. tinctoria were palmitic acid (15.3%), p-cymene (12.6%), and α-muurolene (12.5%), and α-pinene (45.4%), p-cymene (15.5%), and ß-myrcene (13.3%) in aerial parts of A. sylvestris, while isocaryophyllene oxide (31.9%), α-bisabolol (17.5%), and α-pinene (12.4%) were the main constituents in the roots. The most abundant phenolic compounds in aerial parts were the derivatives of caffeic acid, quinic acid, and quercetin; the main compounds in roots of A. sylvestris were chlorogenic acid, quinic acid, and naringenin. The strongest anticancer effects were observed in essential oils of A. sylvestris roots and aerial parts on human carcinoma in the mouth cells (KB, IC50 19.73 µg/mL and 19.84 µg/mL, respectively). The essential oil of A. tinctoria showed a strong effect on KB and LNCaP cells (27.75-29.96 µg/mL). The methanolic extracts of both plants had no effect on the cancer cells studied.

Contribution of branched chain amino acids to energy production and mevalonate synthesis in cancer cells.

Leucine, isoleucine and valine, known as branched chain amino acids (BCAAs), have been reported to be degraded by different cancer cells, and their biodegradation pathways have been suggested as anticancer targets. However, the mechanisms by which the degradation of BCAAs could support the growth of cancer cells remains unclear. In this work, 13C experiments have been carried out in order to elucidate the metabolic role of BCAA degradation in two breast cancer cell lines (MCF-7 and BCC). The results revealed that up to 36% of the energy production via respiration by MCF-7 cells was supported by the degradation of BCAAs. Also, 67% of the mevalonate (the precursor of cholesterol) synthesized by the cells was coming from the degradation of leucine. The results were lower for BCC cells (14 and 30%, respectively). The non-tumorigenic epythelial cell line MCF-10A was used as a control, showing that 10% of the mitochondrial acetyl-CoA comes from the degradation of BCAAs and no mevalonate production. Metabolic flux analysis around the mevalonate node, also revealed that significant amounts of acetoacetate are being produced from BCAA derived carbon, which could be at the source of lipid synthesis. From these results we can conclude that the degradation of BCAAs is an important energy and carbon source for the proliferation of some cancer cells and its therapeutic targeting could be an interesting option.

The Potential of Dietary Antioxidants from a Series of Plant Extracts as Anticancer Agents against Melanoma, Glioblastoma, and Breast Cancer.

In modern society, cancer is one of the most relevant medical problems. It is important to search for promising plant raw materials whose extracts have strong antioxidant and anticancer effects. The aim of this study was to determine the composition of phenolic compounds in plant extracts, to evaluate their antioxidant and anticancer activity, and to find the correlations between those activities. Extracts of calendula, sage, bearberry, eucalyptus, yarrow, and apple were selected for the study. The phenolic compounds of these extracts were determined by the UPLC-ESI-MS/MS method and the antioxidant activity was evaluated in vitro by four different UV-VIS spectrophotometric methods (ABTS, DPPH, CUPRAC, FRAP). The anticancer activity of extracts was tested against melanoma IGR39, glioblastoma U-87, and triple-negative breast cancer MDA-MB-231 cell lines in vitro by MTT assay. The highest content of identified and quantified phenolic compounds was found in sage leaf extract and the lowest in ethanol eucalyptus leaf extract. The highest antioxidant activity was determined by all applied methods for the acetone eucalyptus leaf extract. The majority of extracts were mostly active against the melanoma IGR39 cell line, and possessed the lowest activity against the glioblastoma U-87 cell line. Acetone extract of eucalyptus leaf samples exhibited the highest anticancer activity against all tested cell lines. Strong and reliable correlation has been found between antioxidant and anticancer activity in breast cancer and glioblastoma cell lines, especially when evaluating antioxidant activity by the FRAP method.

Comparison of Ethanolic and Aqueous-Polyethylenglycolic Propolis Extracts: Chemical Composition and Antioxidant Properties.

In recent years, particular attention has been paid to the natural antioxidants. Bee products, especially propolis, are characterized by multifunctional (antioxidant, anti-inflammatory, antibacterial, antiviral, and food preservative) effects and can be used for the development of functional food or food preservatives. Propolis extracts that are commonly produced are ethanolic; therefore, to certain groups of consumers, for example, children and alcohol sensitive group, their applicability is limited. The aim of this study was to develop alternative propolis from aqueous-polyethylenglycolic propolis extract (AQUA-PEG) and compare the chemical composition as well as antioxidant (radical-scavenging and reduction properties) activities to those of ethanolic propolis extract (EEP). Polyethylene glycol is quite a good solvent, which can be successfully used for the preparation of NEP. The total quantity of phenolic compounds identified in AQUA-PEG (400.36 µg/mL), prepared according to our technology, is very similar to that of EEP (433.53 µg/mL), whereas the amount of phenolic acids was greater by 1.31-fold in AQUA-PEG and of flavonoids was greater by 2.38-fold in EEP. The antioxidant activity depends on the method used: by applying the ABTS and CUPRAC methods, both extracts demonstrate similar antioxidant (antiradical and reducing) activity, whereas in the case of the DPPH and FRAP method, significantly higher antioxidant activity was detected in EEP. This should be taken by researchers into account especially when interpreting the results and drawing conclusions about the antioxidant activity of propolis extracts. On the basis of the results, AQUA-PEG, prepared by the developed technology, can be used as an alternative form to ethanolic propolis extract, since it contains a large quantity of antioxidants, namely, flavonoids and phenolic acids. We believe that nonethanolic propolis extract has the prospect of being applied for the development of functional foods in order to alleviate certain symptoms of oxidative stress or for the prevention of some oxidative-stress-related diseases.

Investigation of Phenolic Composition and Anticancer Properties of Ethanolic Extracts of Japanese Quince Leaves.

Glioblastoma multiforme is an aggressive and invasive disease with no efficient therapy available, and there is a great need for finding alternative treatment strategies. This study aimed to investigate anticancer activity of the extracts of the Japanese quince (JQ) cultivars 'Darius', 'Rondo', and 'Rasa' leaf extracts on glioblastoma C6 and HROG36 cells. As identified by ultra high performance liquid chromatography electrospray ionization tandem mass spectrometry, the extracts contained three prevailing groups of phenols: hydroxycinnamic acid derivatives; flavan-3-ols; and flavonols. Sixteen phenols were detected; the predominant compound was chlorogenic acid. The sum of detected phenols varied significantly between the cultivars ranging from 9322 µg/g ('Rondo') to 17,048 µg/g DW ('Darius'). Incubation with the extracts decreased the viability of glioblastoma HROG36 cells with an efficiency similar to temozolomide, a drug used for glioblastoma treatment. In the case of C6 glioblastoma cells, the extracts were even more efficient than temozolomide. Interestingly, primary cerebellar neuronal-glial cells were significantly less sensitive to the extracts compared to the cancer cell lines. The results showed that JQ leaf ethanol extracts are rich in phenolic compounds, can efficiently reduce glioblastoma cell viability while preserving non-cancerous cells, and are worth further investigations as potential anticancer drugs.

Chemical composition, antioxidant, antimicrobial and antiproliferative activities of the extracts isolated from the pomace of rowanberry (Sorbus aucuparia L.).

In the present work, acetone, ethanol and water extracts of rowanberry (Sorbus aucuparia L.) pomace were evaluated for their antiproliferative, antimicrobial and antioxidative effects. Chemical composition of the extracts was determined by UPLC-ESI-MS/MS and spectrophotometric methods. Neochlorogenic and chlorogenic acids were the major phenolic compounds. The water extract contained the highest total proanthocyanidins content (301 ± 18.9 mg/g) and demonstrated the highest antioxidant activity in the all assays (DPPH, FRAP and ORAC). Extracts isolated from rowanberry pomace effectively inhibited the growth of undesirable microorganisms, especially Gram-positive bacteria. Acetone extract was the strongest antimicrobial agent followed by water and ethanol extracts. Acetone and water extracts demonstrated also higher cytotoxic potential in cell viability assays (SRB and MTT) using Caco-2 cells. In general, the results suggest that rowanberry pomace is a promising source of natural compounds with antioxidant and biological activities.

Fatty Acid Synthesis and Degradation Interplay to Regulate the Oxidative Stress in Cancer Cells.

Both cytosolic fatty acid synthesis (FAS) and mitochondrial fatty acid oxidation (FAO) have been shown to play a role in the survival and proliferation of cancer cells. This study aimed to confirm experimentally whether FAS and FAO coexist in breast cancer cells (BCC). By feeding cells with 13C-labeled glutamine and measuring labeling patterns of TCA intermediates, it was possible to show that part of the cytosolic acetyl-CoA used in lipid synthesis is also fed back into the mitochondrion via fatty acid degradation. This results in the transfer of reductive potential from the cytosol (in the form of NADPH) to the mitochondrion (in the form of NADH and FADH2). The hypothesized mechanism was further confirmed by blocking FAS and FAO with siRNAs. Exposure to staurosporine (which induces ROS production) resulted in the disruption of simultaneous FAS and FAO, which could be explained by NADPH depletion.

Hilic MS/MS determination of amino acids in herbs of Fumaria schleicheri L., Ocimum basilicum L., and leaves of Corylus avellana L.

The aim of research was to study the content of amino acids using in extracts of Fumaria schleicheri L., Ocimum basilicum L., and Corylus avellana L. by HILIC MS/MS method. Separation of amino acids in the samples was carried out with Acquity H-class UPLC system (Waters, Milford, USA) equipped with SeQuant ZIC-Hilic collumn (2.1 × 150 mm, 3.5 µm) (Merck Millipore, Darmstadt, Germany). The MS/MS fragment ion chromatograms of the test solutions established the presence of 19 amino acids. The obtained results have shown that O. basilicum L. characterized the highest concentrations of different neurogenic amino acids (128.1 mg/kg), comparing with F. schleicheri L. and C. avellana L. (57.72 and 52.91 mg/kg, respectively).

Antioxidant activity, neuroprotective properties and bioactive constituents analysis of varying polarity extracts from Eucalyptus globulus leaves.

Eucalyptus globulus is employed as herbal tea and therapeutical purposes. In this work, it is investigated for first time the neuroprotective activities, based on antioxidant properties, of varying polarity extracts (acetone, ethanol and methanol) from E. globulus leaves and elucidate their main bioactive constituents. Methanol and acetone extracts contained the highest phenolic compounds amount and chlorogenic acid was the major compound identified by UPLC-ESI-MS/MS. Moreover, the three tested extracts showed significant antioxidant properties, varying their potency depending on the in vitro technique used. Furthermore, E.globulus extracts were effective in ameliorating H2O2-induced oxidative stress by increasing cell viability, GSH levels and antioxidant enzymes activity and, by decreasing ROS production and lipid peroxidation levels in SH-SY5Y cells. Taken together, E.globulus leaves extracts could be used as raw material for food and pharmaceutical supplements for their high content in antioxidant compounds with health benefits properties against oxidative stress.

The impact of CYP2C19*2, CYP4F2*3, and clinical factors on platelet aggregation, CYP4F2 enzyme activity, and 20-hydroxyeicosatetraenoic acid concentration in patients treated with dual antiplatelet therapy.

: The aim of the current study was to evaluate the impact of CYP2C192 (rs4244285), CYP4F23 (rs2108622), and nongenetic factors on platelet aggregation and to investigate the mechanism of CYP4F2's effect on platelet aggregation in the patients treated with dual antiplatelet therapy. A total of 146 patients were included in this study. Ticagrelor or clopidogrel were administered in a loading dose of 180 mg and 600 mg, respectively, in combination with aspirin (300 mg). Blood samples for analysis were taken the next morning after antiplatelet therapy induction. Clopidogrel users with the CYP2C1912 variant had higher platelet aggregation values (median 43, range 30-54%) compared with 11 wild-type carriers (median 33, range 15-77%; P = 0.009). Carriers of the CYP4F213 variant had higher platelet aggregation values than carriers of the 33 variant (median 34, range 8-70% vs. median 24.5, range 10-47%, P = 0.016, respectively). Higher CYP4F2 concentrations were detected in clopidogrel users than in ticagrelor users (median 3.6, range 1.6-22.0 ng/ml vs. median 2.3, range 1.6-27.2 ng/ml, P = 0.056, respectively) and in carriers of the CYP4F213 variant compared with carriers of the 11 variant (median 4.3, range 1.6-27.2 ng/ml vs. median 2.4, range 1.6-22.0 ng/ml, P = 0.009, respectively). No correlation between plasma 20-hydroxyeicosatetraenoic acid and CYP4F2 enzyme concentrations were detected (r = -0.045, P = 0.587). Our results proved that CYP2C192 might significantly affect antiplatelet function of clopidogrel. Plasma CYP4F2 concentrations were significantly lower in ticagrelor users than in clopidogrel users.

Evaluation of phenolic antioxidant content in organically and conventionally grown buckwheat herb crop and its regrowth.

BACKGROUND: Buckwheat herb is known in the pharmaceutical industry as a material rich in phenolics. Buckwheat is also capable of regrowing and producing an additional harvest in the same year. Although buckwheat herb is a popular material, it is not known whether the regrowth has features of the same quality as the first harvest. Therefore, using the herb of 15 varieties of buckwheat cultivated in Lithuania, the present study aimed to examine biometric properties, phenolic content and antioxidant activity as material quality indicators for buckwheat herb and its regrowth under organic and conventional farming conditions. RESULTS: The highest amount of rutin was indicated in the organically cultivated regrowth of 2015. Buckwheat material accumulated significantly (P < 0.05) lower amounts of other phenolics. A significant correlation between phenolic content and antioxidant activity was observed in the herb and its regrowth in both farming systems. CONCLUSION: Regrowth was of similar quality to the first harvest in terms of phenolic content and antioxidant activity, although a downward trend in several biometric features was observed: the height of the regrowth plants was up to two-fold smaller and biomass was five- to nine-fold lower compared to that of the first harvest plants. Organic and industrial farming systems were determined to produce plants of similar quality. © 2016 Society of Chemical Industry.