Radioprotective effects of hawthorn against genotoxicity induced by gamma irradiation in human blood lymphocytes

The radioprotective effect of hawthorn (Crataegus microphylla) fruit extract was investigated in cultured blood lymphocytes from human volunteers. Peripheral blood samples were collected from five human volunteers 10 min before and 1, 2 and 3 h after a single oral ingestion of 500 mg hawthorn powder extract. At each time point, the whole blood was exposed in vitro to 150 cGy of cobalt-60 gamma irradiation, and then the lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis-blocked binucleated cell. The lymphocytes in the blood samples collected after extract ingestion exhibited a significant decrease in the incidence of binucleated cells containing micronuclei as compared to similarly irradiated lymphocytes collected prior to extract ingestion. The maximum decrease in the frequency of micronuclei-containing cells was observed at 1 h after ingestion of Hawthorn extract (on average a 44% decrease). These data suggest that it may be possible to use Hawthorn extracts in personnel exposed to radiation in order to protect lymphocytes from radiation effects.     

Pharmacology of Acorus calamus L

Water soluble dried powder of alcoholic extract of roots and rhizomes of A. calamus L. was used. The in vivo experiments involved strychnine convulsant activity in frogs, spontaneous motor activity and amphetamine hyperactivity in mice, pentobarbitone sleeping-time in rats and local anaesthetic activity in guinea pigs and rabbits. Frog skeletal muscle and heart preparations and rat phrenic nerve diaphragm constituted the in vitro experiments. Plant extracts at 10, 20 mg/kg ip did not afford protection to strychnine (1,5,2.5 mg/kg) induced convulsions and same effect was found on acetylcholine induced contractions of rectus muscle except that it inhibited caffeine citrate contractions in frog. At 1, 10 and 100 micrograms/ml doses, it caused negative iono- and chronotropic effects in frogs. Dosages of 10, 25, 50 mg/kg ip of herbal extract antagonize spontaneous motor activity and also amphetamine induced hyperactivity in mice. It was less potent than chloropromazine, though exerts sedative and tranquilizing action. Local anaesthetic activity was found to be absent at 0.5 and 1% dose levels.     

Polyphenolic content and physiological activities of Chinese hawthorn extracts

Hawthorn polyphenol (HP) was prepared by ethyl acetate treatment of the ethanol extract (HE) of Chinese hawthorn fruit. The concentrations of 15 polyphenols in the HP, HE, extraction residue (HJ), and a hawthorn leaf extract (HF) were determined by HPLC. For HP, the total content of the 15 polyphenols was 21.4%, comprised of 19.7% of procyanidins, 1.21% of chlorogenic acid, and 0.48% of flavonoids, compared to 2.55% for the HE. The yields of procyanidin monomer, dimer, trimer, tetramer, and pentamer were 50.5%, 30.3%, 23.0%, 14.6%, and 12.5% respectively, and the mean degree of polymerization was reduced to 1.39 (HP) from 1.65 (HE). Seven different physiological actions of the four extracts were investigated. The HP showed strong O(2)(-) and (.-)OH scavenging capacities (IC(50) values of 6.3 microg/ml and 1.1 microg/ml respectively), as well as selective prolyl endopeptidase inhibition (IC(50) of 60 microg/ml). The active constituents appeared to be procyanidins.     

Polyphenolic Content and Physiological Activities of Chinese Hawthorn Extracts

Hawthorn polyphenol (HP) was prepared by ethyl acetate treatment of the ethanol extract (HE) of Chinese hawthorn fruit. The concentrations of 15 polyphenols in the HP, HE, extraction residue (HJ), and a hawthorn leaf extract (HF) were determined by HPLC. For HP, the total content of the 15 polyphenols was 21.4%, comprised of 19.7% of procyanidins, 1.21% of chlorogenic acid, and 0.48% of flavonoids, compared to 2.55% for the HE. The yields of procyanidin monomer, dimer, trimer, tetramer, and pentamer were 50.5%, 30.3%, 23.0%, 14.6%, and 12.5% respectively, and the mean degree of polymerization was reduced to 1.39 (HP) from 1.65 (HE). Seven different physiological actions of the four extracts were investigated. The HP showed strong O₂ ^? and ^?OH scavenging capacities (IC₅₀ values of 6.3 μg/ml and 1.1 μg/ml respectively), as well as selective prolyl endopeptidase inhibition (IC₅₀ of 60 μg/ml). The active constituents appeared to be procyanidins.     

Ligand-independent negative chronotropic responses of rat and mouse right atria to beta-adrenoceptor antagonists

Negative chronotropic effects of beta-adrenoceptor (betaAR) antagonists on right atria from reserpine-treated rats and mice were determined as a test of their inverse agonist activities. BetaAR antagonist ICI-118,551 and nonselective betaAR antagonists alprenolol, propranolol, and timolol produced negative chronotropic effects. In contrast, nonselective betaAR antagonists pindolol and nadolol as well as beta1AR-selective antagonists atenolol, acebutolol, and metoprolol did not cause a significant decrease in atrial rates. The neutral antagonist pindolol but not the inverse agonist alprenolol inhibited the negative chronotropic activities of ICI-118,551. Isoprenaline, salbutamol, and noradrenaline produced positive chronotropic effects; the chronotropic effects of isoprenaline and salbutamol but not of noradrenaline were antagonized by ICI-118,551. It is concluded that both beta1AR and beta2AR mediate positive chronotropic effects of catecholamines on rat and mouse atria but only beta2AR are constitutively active.     

Effects of hirsutine and dihydrocorynantheine on the action potentials of sino-atrial node, atrium and ventricle

The effects of hirsutine, an indole alkaloid from Uncaria rhynchophylla MIQ. JACKSON with antihypertensive, negative chronotropic and antiarrhythmic activity, and its C3 structural epimer, dihydrocorynantheine, on membrane potentials of rabbit sino-atrial node and guinea-pig right ventricle and left atrium were studied with microelectrode techniques. In sino-atrial node preparations, hirsutine and dihydrocorynantheine (0.1 microM to 10 microM) concentration-dependently increased cycle length, decreased slope of the pacemaker depolarization (phase 4 depolarization), decreased maximum rate of rise and prolonged action potential duration. In atrial and ventricular preparations, both compounds (0.1 microM to 30 microM) concentration-dependently decreased maximum rate of rise and prolonged action potential duration. These results indicate that hirsutine and dihydrocorynantheine have direct effects on the action potential of cardiac muscle through inhibition of multiple ion channels, which may explain their negative chronotropic and antiarrhythmic activity.     

Cardioactive properties of Solanaceae plant extracts and pure glycoalkaloids on Zophobas atratus

Glycoalkaloids, the biologically active secondary metabolites produced by Solanaceae plants, are natural defenses against animals, insects and fungi. In this paper, the effects of glycoalkaloids present in extracts of Solanaceae plants (potato, tomato and black nightshade) or pure commercial glycoalkaloids on the coleopteran Zophobas atratus F. were evaluated by in vitro and in vivo bioassays using heart experimental models. Each tested extract induced a dose‐dependent cardioinhibitory effect. The perfusion of Zophobas atratus semi‐isolated heart using the highest potato and tomato extract concentration (1 mmol/L) caused irreversible cardiac arrests, while extract from black nightshade produced fast but reversible arrests. Pure commercial glycoalkaloids caused similar but less evident effects compared with extracts. Our results showed that the bioactivity of tested compounds depended on their structure and suggested the existence of synergistic interactions when combinations of the main glycoalkaloids of potato and black nightshade were used for trials. Surprisingly, injection of tomato and potato extracts in 1‐day‐old pupae of Zophobas atratus induced reversible positive chronotropic effects and decreased the duration of the both phases (anterograde and retrograde) of the heart contractile activity. Furthermore, these extracts affected the amplitude of the heart contractions.     

Allelopathic effects of water hyacinth [Eichhornia crassipes

Eichhornia crassipes (Mart) Solms is an invasive weed known to out-compete native plants and negatively affect microbes including phytoplankton. The spread and population density of E. crassipes will be favored by global warming. The aim here was to identify compounds that underlie the effects on microbes. The entire plant of E. crassipes was collected from El Zomor canal, River Nile (Egypt), washed clean, then air dried. Plant tissue was extracted three times with methanol and fractionated by thin layer chromatography (TLC). The crude methanolic extract and five fractions from TLC (A-E) were tested for antimicrobial (bacteria and fungal) and anti-algal activities (green microalgae and cyanobacteria) using paper disc diffusion bioassay. The crude extract as well as all five TLC fractions exhibited antibacterial activities against both the gram positive bacteria; Bacillus subtilis and Streptococcus faecalis; and the gram negative bacteria; Escherichia coli and Staphylococcus aureus. Growth of Aspergillus flavus and Aspergillus niger were not inhibited by either E. crassipes crude extract nor its five fractions. In contrast, Candida albicans (yeast) was inhibited by all. Some antialgal activity of the crude extract and its fractions was manifest against the green microalgae; Chlorella vulgaris and Dictyochloropsis splendida as well as the cyanobacteria; Spirulina platensis and Nostoc piscinale. High antialgal activity was only recorded against Chlorella vulgaris. Identifications of the active antimicrobial and antialgal compounds of the crude extract as well as the five TLC fractions were carried out using gas chromatography combined with mass spectroscopy. The analyses showed the presence of an alkaloid (fraction A) and four phthalate derivatives (Fractions B-E) that exhibited the antimicrobial and antialgal activities.     

Investigation of the pharmaceutical and pharmacological equivalence of different Hawthorn extracts

Seven Hawthorn extracts were tested in isolated guinea pig aorta rings. The effect on noradrenaline- (10 microM) induced contraction was investigated. The extracts were prepared using ethanol (40 to 70% v/v), methanol (40 to 70% v/v), and water as the extraction solvents. The aqueous-alcoholic extracts displayed similar spectra of constituents. They were characterised by similar procyanidin, flavonoid, total vitexin and total phenols content and by similar TLC fingerprint chromatograms. The aqueous extract, however, showed a different fingerprint and a noticeably lower concentration of procyanidins, flavonoids and total phenols but a similar total vitexin content. All 7 extracts had a relaxant effect on the aorta precontracted by noradrenaline and led to relaxations to 44 until 29% of the initial values. The EC50 values of the aqueous-alcoholic extracts varied between 4.16 and 9.8 mg/l. The aqueous extract produced a similarly strong maximal relaxation as the other extracts, but the EC50, at 22.39 mg/l, was markedly higher. The results show that Hawthorn extracts with comparable quality profiles were obtained by using aqueous-alcoholic extraction solvents (40 to 70% ethanol or methanol). The extracts exerted comparable pharmacological effects. When using water as the extraction solvent, both, the spectrum of constituents and the pharmacological effect, deviated remarkably. It is thus possible to obtain bioequivalent extracts with comparable effect profiles by using 40 to 70% ethanol or methanol as the extraction solvent.     

The roles of alpha- and beta-adrenoceptors in the chronotropic responses to norepinephrine in carp heart (Cyprinus carpio)

1. The chronotropic effect of norepinephrine was studied in isolated spontaneously beating atrial preparations of carp (Cyprinus carpio) heart. 2. Norepinephrine, 0.1 microM, caused a positive chronotropic effect, while at 1 microM it caused either a positive or a negative chronotropic effect. The positive chronotropic effect, observed in 13 preparations, was potentiated by phentolamine and almost completely blocked by propranolol. 3. The negative chronotropic effect observed in the other 5 preparations was greater in the presence of propranolol, reduced by phentolamine and not affected by atropine. 4. These results indicate that alpha- and beta-adrenoceptors may coexist, mediating the negative and positive chronotropic effects, respectively, in isolated atrial preparations of carp heart.     

Cardiovascular effects of atrial natriuretic extract in the whole animal

Atrial tissue extract (AE) and ventricular tissue extract cause identical decreases in total peripheral resistance when they are injected i.v. into anesthetized rats. However, only AE causes significant hypotension because of cardiac inhibition. This involves both bradycardia and failure of stroke volume to increase appropriately. The observations cannot be explained by direct action of AE on myocytes, but are more likely to be the result of interactions with cardiovascular reflex mechanisms. Excitation of chemosensitive cardiac receptors with vagal afferents appears to be an important afferent mechanism. The efferent limb for the negative chronotropic response resides partly in the vagus nerves and partly in cardiac sympathetic nerves. The negative inotropic response of AE was not altered by vagotomy, spinal section, atropine, or propranolol. These results suggest that atrial peptides may cause the release of a negatively inotropic substance from a site that is not yet identified.     

Chemical characterisation,antioxidant and antimicrobial screening for the revaluation of wine supply chain by-products oriented to circular economy

Abstract

Aim of the project was the bioassay guided optimisation of extraction methods applied to wine chain by-product to obtain extracts, fractions and biologically active biomolecules with a possible use in the nutraceutical and cosmeceutical industry. Exhausted red and white grape marc were extracted using water:ethanol 50:50 with ultrasound assisted extraction and Naviglio® technology; and also with supercritical fluid extraction (SFE) and steam-distillation obtaining different phytocomplexes. Each extract was characterised by different molecular category: exhausted red grape marc (VCR) by anthocyanins, exhausted white grape marc (VCB) by flavonoids, and grapeseed (VIN) by proanthocyanins. SFE and steam-distillation, instead, highlighted the presence of fatty acids and their ester in every matrix, but terpenoids were not revealed at level major or equal than 0.1%, except for manoyl oxide in VCR (2.89%). VIN was the most abundant matrix in polyphenols (506.24?±?55.91?mg gallic acid/g dried extract), and it showed the highest antioxidant activity (IC₅₀ of 4.30?μg/mL). Regarding the antimicrobial activity, the hydroalcoholic extracts from VCR, VCB and VIN were tested but no noteworthy activities have been recorded.     

Purification of Thiol-disulfide Interchange Enzyme from Candida claussenii

Thiol-disulfide interchange enzyme which catalyzes the thiol-disulfide interchange was purified from cell-free extracts of Candida claussenii by acid treatment, ammonium sulfate fractionation, aqueous polymer two phase method (Dextran-PEG system), CM-Sephadex column chromatography, Sephadex G–100 and Sephadex G–200 gel filtrations. More than four active fractions were obtained on CM-Sephadex column. Further purification steps from one of these fractions resulted in two purified enzyme preparations D–l–1 and D–2 of which the increase in specific activities was 8150- and 8450-folds respectively, over the crude extract. Both purified enzymes were homogeneous in ultracentrifugal analysis.     

Anti-Helicobacter pylori activities of six Iranian plants

BACKGROUND: Helicobacter pylori is the major worldwide cause of bacterial gastrointestinal infections in adults and children. Antibiotic therapy and a combination of two or three drugs have been widely used to eradicate these infections. However, development of drug resistance in bacteria calls for new sources of drugs, and plants seem to be a logical source of new antibacterial compounds. METHODS: The anti-H. pylori activities of six native Iranian plants (Glycyrrhiza aspera, Juglans regia, Ligustrum vulgare, Thymus kotschyanus, Trachyspermum copticum and Xanthium brasilicum) and seven antibiotics were determined against 70 clinical isolates from children using the disk susceptibility assay. Minimum inhibitory concentrations were also measured for the biologically active extracts. One extract with the best anti-H. pylori activity was fractionated by silica gel and thin layer chromatography and the active compounds were identified by hydrogen nuclear magnetic resonance ((1)HNMR) spectroscopy. RESULTS: All plant extracts showed anti-H. pylori activity by the disk sensitivity method, but the most active extracts were those from X. brasilicum and T. copticum. In fact, the anti-H. pylori activities of the two extracts were superior to the disk antibiotic susceptibility profile. Minimum inhibitory concentrations were within the range of 31.25-250 micro g/ml. Fractionation and chemical identification of the extract from X. brasilicum showed the presence of two substances, a flavonoid and a xanthanolide. CONCLUSIONS: Due to the rise in antibiotic resistance, new sources of anti-H. pylori drugs are needed. The use of medicinal plants and/or their chemical components may have potential benefit in eradicating such problems.     

Characterization of genital tract growth factor-like activity from testis sheaths and fat body of Heliothis virescens

Testis sheaths and fat body from developing male pupae of Heliothis virescens synthesize soluble growth factor-like products when exposed to 20-hydroxyecdysone (20HE). These factors promote growth and development of the genital tract. Saline extracts of modified Grace's medium and 20HE-treated testis sheaths and fat body were subjected to heat, freeze-thaw, organic solvents, and low pressure size exclusion chromatography. Although extracts were stable to repeated freeze-thawing, activity was lost after exposure to organic solvents; activities of fractions heavier than 6.5 KDa were inhibited by heating to 100°C. Size exclusion chromatography yielded 10 active testis extract fractions and 9 active fat body fractions. Although the approximate molecular weights of most of the extract fractions were similar, enzyme studies using protease, lipase, and α-amylase indicated differences in the chemistry of active fractions derived from the two tissues. Active factors were inhibited by protease or lipase or both enzymes. © 1994 Wiley-Liss, Inc.     

Delayed hypersensitivity to crude and partially purified murine tumor-specific transplantation antigens and alloantigens utilizing the footpad swelling assay

Antigen preparations extracted from C3H/HeJ methylcholanthrene-induced fibrosarcomas by the 3M KCl extraction procedure were assessed for tumor-specific and allospecific antigenicity. Specificity of crude tumor antigen preparations and of fractions from preparative isoelectric focusing was investigated by evocation of footpad swelling (FPS) in syngeneic mice immunized with irradiated fibrosarcoma cells. Tumor immune mice displayed delayed hypersensitivity as positive FPS responses to challenge with 3M KCl extract and with fraction (Fr) 15 (pH 5.7 to 6.0) from preparative isoelectrically focused 3M KCl extract. Crude extracts and Fr 15 exhibited immunoprotective activity in vivo. Immune mice demonstrated a specific FPS response only to crude antigen preparations of Fr 15 from immunizing tumors, not to materials from a noncross-reactive neoplasm. DBA/2J mice immunized with C3H/HeJ spleen cells displayed FPS to challenge with crude antigen preparations, but not with the tumor-specific Fr 15. Alloantigen activity, however, was detected by a positive FPS response in C3H-immune DBA mice in fractions from the pH range 5.1 to 5.5. These experiments demonstrated that the FPS assay provides the setting for detection of specific delayed hypersensitivity responses to crude and fractionated tumor antigen preparations and for delineation of tumor-specific and histocompatibility antigen activities in fractions from crude extracts.     

Cardiovascular effects in vitro of aqueous extract of wild strawberry (Fragaria vesca,L.) leaves

In contrast to the strawberry fruits, strawberry leaves as a source of bioactive compounds with potentially beneficial biological effects have been largely overlooked. In this study we examined direct, dose-dependent effects of wild strawberry (Fragaria vesca, L.) leaves aqueous extract, in two experimental models and animal species, the isolated guinea pig hearts and rat aortic rings. Vasodilatory potential of the wild strawberry leaves extract was compared with vasodilatory activity of aqueous extract of hawthorn (Crataegus oxycantha, L) leaves with flowers, which can be regarded as a reference plant extract with a marked vasodilatory activity.The extracts were analysed by their “phenolic fingerprints”, total phenolic content and antioxidative capacity. Their vasodilatory activity was determined and compared in the isolated aortic rings from 24 rats that were exposed to the extracts doses of 0.06, 0.6, 6, and 60 mg/100 ml. Both extracts induced similar, dose-dependent vasodilation. Maximal relaxation was 72.2±4.4% and 81.3±4.5%, induced by the strawberry and hawthorn extract, respectively. To determine vasodilatory mechanisms of the wild strawberry leaves extract, endothelium-denuded and intact rings exposed to nitric oxide (NO) synthase inhibitor l-NAME or cyclooxygenase inhibitor indomethacin were used. Removal of the endothelium prevented and exposure to l-NAME or indomethacin strongly diminished the vasodilatatory response to the extract. In the isolated hearts (n=12), the wild strawberry extract was applied at concentrations of 0.06, 0.18, 0.6, and 1.8 mg/100 ml. Each dose was perfused for 3.5 min with 15 min of washout periods. Heart contractility, electrophysiological activity, coronary flow and oxygen consumption were continuously monitored. The extract did not significantly affect heart rate and contractility, main parameters of the cardiac action that determine oxygen demands, while coronary flow increased up to 45% over control value with a simultaneous decrease of oxygen extraction by 34%.The results indicate that the aqueous extract of wild strawberry leaves is a direct, endothelium-dependent vasodilator, action of which is mediated by NO and cyclooxygenase products and which potency is similar to that of the hawthorn aqueous extract.     

Bacopa monniera, a reputed nootropic plant: an overview

Bacopa monniera (BM), a traditional Ayurvedic medicine, used for centuries as a memory enhancing, anti-inflammatory, analgesic, antipyretic, sedative and antiepileptic agent. The plant, plant extract and isolated bacosides (the major active principles) have been extensively investigated in several laboratories for their neuropharmacological effects and a number of reports are available confirming their nootropic action. In addition, researchers have evaluated the anti-inflammatory, cardiotonic and other pharmacological effects of BM preparations/extracts. Therefore, in view of the important activities performed by this plant, investigation must be continued in the recently observed actions described in this paper. Moreover, other clinical studies have to be encouraged, also to evidence any side effects and possible interactions between this herbal medicine and synthetic drugs.     

香椿嫩芽和老叶萃取物抗菌活性的比较研究

采用琼脂扩散法对香椿嫩芽和老叶萃取物的氯仿、乙酸乙酯和正丁醇萃取部分的抗菌活性进行了研究.嫩芽萃取物的抗菌活性明显高于老叶萃取物,其中嫩芽萃取物的乙酸乙酯部分表现出最强的抗菌活性,其最小抑菌浓度(MIC)为1.3 mg/mL.实验结果表明,香椿叶中的抑菌物质随着生长时间的延长而逐渐降低.     

Activity of Hawthorn Leaf and Bark Extracts in Relation to Biological Membrane

The aim of the study was to identify and determine the percent content of polyphenols in extracts from leaves and hawthorn bark, to examine the effect of the extracts on the properties of the biological membrane as well as to determine their antioxidant activity toward membrane lipids. In particular, a biophysical investigation was conducted on the effect of hawthorn extracts on the osmotic resistance and morphology of erythrocyte cells and on the packing of the heads of membrane lipids. Analysis of the polyphenol content of extracts used the HPLC method. Analysis of the polyphenol composition has shown a dominant share of procyanidins and epicatechin in both extracts. The research showed that the polyphenolic compounds contained in hawthorn extracts are incorporated mainly into the hydrophilic part of the erythrocyte membrane, inducing echinocyte shapes. They also diminish the packing order of the lipid polar heads of the membrane, as evidenced by the lowered generalized polarization values of Laurdan. The substances used induced increased osmotic pressure of erythrocytes, making them less sensitive to changes in osmotic pressure. The presence of the extract compounds in the outer hydrophilic part of the erythrocyte membrane, evidenced by examination of the shapes and packing in the hydrophilic part of membrane, indicates that the substances constitute a kind of barrier that protects the erythrocyte membrane against free radicals, while the membrane-bound extracts do not disturb the membrane structure and, thus, do not cause any side effects.