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Effect of various pretreatments on the hypothermic activity of repin in naive rats.

Repin is a sesquiterpene lactone found in Centaurea solstitialis, a plant responsible for Parkinson-like disease in horses. Repin, on intraperitoneal (i.p.) injection, produces a dose-dependent and highly significant hypothermia in naive rats. The hypothermia is long-lasting with a peak 3 h after the injection and a return to normal temperature after more than 8 h. The effects of atropine sulfate, atropine methylbromide, propranolol, metergoline, ketanserin, diphenhydramine and apomorphine pretreatment on repin-induced hypothermia were investigated. None of the pretreatments directly antagonized repin's hypothermic effect. However, partial but significant reversals of hypothermia by atropine sulfate, metergoline, ketanserin, diphenhydramine and apomorphine were observed 2-4 h after the repin injection. These late-onset effects are probably due to secondary physiological mechanisms. On the other hand, propranolol, at 20 mg/kg i.p., clearly accentuated both the early-onset (30-90 min) and late-onset (2-4 h) hypothermic effects of repin.

Animals↗

Traditional phytotherapy in Central Italy (Marche, Abruzzo, and Latium).

In this study, the more significant results of extensive ethnopharmacobotanical research carried out by the author in the years 1977-2000 in 175 localities of three regions of Central Italy (Marche, Abruzzo, and Latium) have been reported and compared. The usages of 80 species belonging to 36 families are described, of which 71 were used in human therapy and 29 in veterinary medicine. Uses are suited with the number of localities in which they have been mentioned. Among the wild plant mainly still used, Malva sylvestris, Urtica dioica, and Sambucus nigra are particularly highly considered, while major uses of plants concern these plants in addition to Allium sativum, Rubus ulmifolius, Parietaria diffusa, Cynodon dactylon, and Ficus carica. Unusual phytotherapic uses concern Brassica oleracea, Taraxacum officinale (warts), Ruscus aculeatus, S. nigra (chilblains), Allium cepa (chilblains; to remove thorns and splinters), Juglans regia, R. ulmifolius (burns), and Euphorbia paralias (bites of weevers). Among new uses with only one quotation, we remember Cirsium arvense (intestinal disturbances), Centaurea bracteata (cough), Lupinus albus (calluses), Melittis melissophyllum (eye inflammations, antispasmodic), and Artemisia absinthium (tendon inflammations), while among plants employed in various regions with interesting less-known properties, there are C. arvense (emergency haemostatic), P. diffusa (insect bites), and Scrophularia canina (antiseptic and cicatrizing agent for wounds in bovines and sheep).

Ethnobotany↗

Antimicrobial activity of some endemic plants growing in the Eastern Black Sea Region, Turkey.

The Eastern Black Sea Region has an extensive flora because of ample rainfall lasting all year. In this study, antimicrobial effects of 74 crude extracts of 22 endemic plants were investigated. Among the 30 active crude extracts, water-insoluble crude extracts from Betula medwediewii, Heracleum platytaenium, Primula longipes, Anthemis cretica ssp. argaea and Centaurea helenioides were the prominent ones with their MIC values.

Anti-Bacterial Agents↗

Screening of selected indigenous plants of Lebanon for antimicrobial activity.

The objective of this study is to test in vitro the antimicrobial efficacy of 39 water and 39 methanol extracts derived from different parts of 27 indigenous wild plant species that have been commonly used in Lebanese folk medicine. The antimicrobial efficacy was determined using the single disk diffusion method, with 10 and 20 microl load extract volume per disc. Nine test microorganisms were used namely, Escherichia coli, Proteus sp., Pseudomonas aeruginosa, Shigella dysenteria, Salmonella enteritidis, Salmonella typhi, Staphylococcus aureus, Streptococcus faecalis, and Candida albicans. Only one water extract out of 39 derived from whole plant of Alchemilla diademata showed an antimicrobial activity against Staphylococcus aureus. The percentage of test organisms that were susceptible to 10 most efficacious methanol plant extracts (20 microl/disc) were as follows: Achillea damascena whole plant (88.8%), Anthemis scariosa flower (88.8%), Cirsium sp. whole plant (88.8%), Centaurea ainetensis flowers (88.8%), Hieracium sp. whole plant (88.8%), Origanum libanoticum whole plant (99.9%), Ranunculus myosuroudes whole plant (88.8%), Nepata curviflora leaf (88.8%), Nepata curviflora stem, and Verbascum leptostychum flower (99.9%). The minimum inhibitory concentration (MIC) was determined on plant extracts that showed high efficacy against the test organisms. The chance to find antimicrobial activities was more apparent in methanol rather than water extracts of the same indigenous plants of Lebanon, with higher antimicrobial activities in 20 microl methanol extract-discs in comparison to that present in the 10 microl discs (P < 0.05).

Anti-Infective Agents↗

Characterization of the role of glutathione in repin-induced mitochondrial dysfunction, oxidative stress and dopaminergic neurotoxicity in rat pheochromocytoma (PC12) cells.

Repin, a major constituent in extracts of the plant Centaurea repens is thought to be the active principal responsible for the development of equine nigropallidal encephalomalacia (ENE), a fatal Parkinson-like neurodegenerative disorder in horses. Although the exact mechanism by which ingestion of this weed causes ENE is uncertain, a limited body of experimental evidence suggests a critical role for the glutathione redox system. In the present study, the mechanism of repin neurotoxicity was examined in PC12 cells with a focus on determining the role of glutathione (GSH) in repin-induced mitochondrial dysfunction, oxidative stress and dopaminergic toxicity. The results demonstrate that repin reduced both cellular GSH levels and mitochondrial function in a manner that was time- and concentration-dependent. The repin-induced changes in GSH levels were found to precede the changes in mitochondrial function. Depletion of GSH with a potent GSH depletor (ethacrynic acid (EA)) and a GSH synthesis inhibitor (buthionine sulfoximine (BSO)) prior to repin treatment enhanced the repin-induced mitochondrial change. In addition, repin caused a concentration-dependent decrease in cellular dopamine levels in NGF-differentiated PC12 cells. Increases in intracellular GSH levels induced by pre-treatment with reducing agents (N-acetyl-L-cysteine or reduced glutathione) completely protected the cells from repin-induced mitochondrial and dopaminergic toxicity. Antioxidants, coenzyme-Q and ascorbic acid completely blocked repin-induced dopaminergic toxicity. These data suggest that GSH plays a critical role in repin-induced neurotoxicity and that the maintenance of neuronal redox status may prove to be a useful strategy for the prevention and/or treatment of ENE. The results support the view that GSH depletion, leading to oxidative damage and subsequent mitochondrial dysfunction, may serve as a trigger for neuronal cell death.

Animals↗

Serine proteinase inhibitors in the Compositae: distribution, polymorphism and properties.

Multiple molecular forms of inhibitors of trypsin (TI) and chymotrypsin (CI), which are typical digestive enzymes of insects, mammals and micro-organisms, and subtilisin (SI), a proteinase of many bacteria and phytopathogenic fungi, were identified in seeds and vegetative organs of the majority of 128 wild and cultivated species representing 65 genera of three of the subfamilies of the Compositae. Inhibitors with M(r) ranging from 7450 to 7800 and combining activities towards subtilisin and trypsin and/or chymotrypsin (T/C/SI) had the widest distribution and may be involved in plant defense mechanisms. They were found in many species of the subfamilies Carduoideae (genera Carthamus, Centaurea, Cirsium), Cichorioideae (Lactuca, Taraxacum) and Asteroideae (Helianthus, Cosmos, Bidens). Partial amino acid sequencing showed that the safflower (Carthamus tinctorius) T/C/SI and Cosmos bipinnatus T/C/SI, T/SI and C/SI belonged to the potato I inhibitor family. The most active, variable and heterogeneous inhibitors were found in species of the tribe Heliantheae, which is placed in the evolutionary advanced subfamily Asteroideae. Seeds of Helianthus species, Eclipta prostrata, Gailardia aristata, Zinnia elegans and Silphium perfoliatum contained various TI with M(r) ranging from 1500 to 14,750, with some also containing SI. H. annuus seeds contain a unique cyclic TI of M(r) 1514 and similar TI were also present in other Helianthus spp. and the related species Tithonia diversifolia. Zinnia elegans contained a TI with M(r) 11,350 which appeared to represent a novel type of inhibitor distantly related to the cereal subgroup of Bowman-Birk inhibitors. TI and T/SI varied widely in H. annuus lines and wild Helianthus species in their presence or absence and composition. Similar T/SI components were found in the cultivated diploid H. annuus and annual diploid species with the B genome but not in perennials with the A genome. Some T/SI, SI and TI were detected in vegetative organs of sunflower and other Compositae. Studies of the polymorphism and distribution of proteinase inhibitors are relevant to the evolution of protective protein systems and the mechanisms of resistance to pathogenic organisms in the Compositae and other plants.

Asteraceae↗

Potential bioindicator plant species for ambient ozone in forested mountain areas of central Europe.

From 1993 to 2000, trees, shrubs, forbs and vines were evaluated for symptoms of probable ozone injury in the vicinity of passive ozone samplers or active ozone monitors in forest condition assessment networks in mostly mountainous regions, principally the Carpathian Mountain Range, in the central European countries Czech Republic, Poland, Romania, Slovakia and Ukraine. Each country was visited at least twice during the time period. Over the course of eight seasons, 29 species of native plants were identified as potential bioindicators of ozone. This is the first report of probable ozone injury on native plants in central Europe. Forbs and shrubs made up the bulk of the species (21 of 29). Potential bioindicators that are widely distributed include the forbs Centaurea nigra. and Impatiens parviflora and the shrubs Alnus incana, Corylus avellana, and Sambucus racemosa. Ozone concentrations in forcsted areas of central Europe appear to be high enough and of sufficient duration to cause foliar injury on a wide variety of native plants.

Air Pollutants↗

Ethnopharmacology of liakra: traditional weedy vegetables of the Arbëreshë of the Vulture area in southern Italy.

An ethnobiological field study on food plants and medicinal foods traditionally consumed in three Arbëresh (ethnic Albanian) communities in northern Lucania (southern Italy) document approximately 120 botanical taxa used for these purposes. Non-domesticated food vegetables (liakra), mostly gathered during the spring season, play a central role as traditional functional food. Quantitative ethnobotanical, ethnotaxonomical, ethnoecological, ethnogastronomical, and ethnopharmacological aspects related to gathering, processing, cooking and consumption of liakra are discussed. Unusual food species, such as Lycium europeaum, Centaurea calcitrapa, and a few spontaneous weedy Asteraceae and Brassicaceae species are locally used in the kitchen. Most of these are very poorly known phytochemically and phytopharmacologically. Moreover, an analysis of taste perception of the most commonly used botanical foods was conducted in the village of Ginestra. Arbëresh taste classification and indigenous criteria related to the perception of bitter taste in considering non-cultivated plants as food or medicine are discussed as well.

Cooking↗

Screening seeds of Scottish plants for antibacterial activity.

Based on ethnopharmacological and taxonomic information, seeds of 21 Scottish plant species from 14 different families were obtained from authentic seed suppliers. Their n-hexane, dichloromethane and methanol extracts were assessed for antibacterial activity against 11 pathogenic bacterial species. Methanol extracts of 11 plant species showed significant antibacterial activity. Malva moschata and Prunus padus were active against five bacterial species, Reseda lutea against four, Centaurium erythraea and Crithmum maritimum against three, Calluna vulgaris against two, and Armeria maritima, Centaurea scabiosa, Daucus carota, Rosa canina and Stellaria holostea against one bacterial species. C. erythraea and P. padus were also active against methicillin resistant Staphylococcus aureus.

Anti-Bacterial Agents↗

Medicinal plants used in the Barros Area, Badajoz Province, Spain.

A study of the wild and cultivated medicinal plants used in the Barros Area (southern Spain) is reported, 48 plants distributed among 20 different families are used in the treatment of various human diseases. The use of Bellis annua L. Centaurea ornata Wild., Leuzea conifera (L.) DC., Pulicaria paludosa Link and Asparagus aphyllus L. is reported.

Data Collection↗

Screening of Turkish anti-ulcerogenic folk remedies for anti-Helicobacter pylori activity.

The anti-Helicobacter pylori effect of the extracts and fractions obtained from seven Turkish plants, which are used in folk medicine for the treatment of gastric ailments including peptic ulcers, were studied against one standard strain and eight clinical isolates of H. pylori by using the agar dilution method. Flowers of Cistus laurifolius and Spartium junceum, cones of Cedrus libani, herbs and flowers of Centaurea solstitialis ssp. solstitialis, fruits of Momordica charantia, herbaceous parts of Sambucus ebulus, and flowering herbs of Hypericum perforatum were evaluated in this study. Results showed that all except one extract from six of these plants showed activity against the microorganism with MICs between 1.95 and 250 microg/ml, with S. junceum being the only inactive species. Amongst the active plants the inhibitory properties of C. laurifolius were found prominent.

Anti-Ulcer Agents↗

Extraction and partial characterization of a coagulant preparation from Silybum marianum flowers. Its action on bovine caseinate.

An outstanding parameter in cheese making is the type of coagulant, which greatly influences the characteristics of the final products. Proteolysis is the most important set of biochemical changes during ripening of most cheeses, and is carried out, in different magnitude, by proteolytic agents originated in milk, rennet (or rennet substitute), and starter and non-starter micro-organisms (Silva & Malcata, 2000). The demand for alternative sources of milk coagulants, to replace the expensive and limited natural rennet supplies, has increased (Esteves et al. 2001). All commercial enzymes employed as milk coagulant are aspartic proteinases, which are most active at acidic pH and preferentially cleave peptide bonds between residues with hydrophobic side-chains (Silva & Malcata, 1999). Because of the presence of aspartic proteinases, aqueous crude extracts from flowers of Cynara cardunculus (Veríssimo et al. 1995, 1996), Cynara humilis, and/or Cynara scolymus are traditionally employed in the Iberian Peninsula as vegetable rennet for cheesemaking (Reis et al. 2000). Milk clotting activity was also proved in flowers of Centaurea calcitrapa and Onopordum turcicum (Tamer, 1993; Domingos et al. 1998). All these species are included within the Asteraceae family and furthermore in the same tribe: Cardueae Cass.= Cynareae Less. (Ariza Espinar & Delucchi, 1998). When a potential rennet substitute is studied, it is particularly important to evaluate adequately the degradation patterns of the caseins because of their effects on yield, consistency, and flavour of the final cheese (Fox, 1989). It is important to guarantee a well-balanced breakdown of curd proteins (caseins) in order to avoid formation of undesired attributes in cheese such as low viscosity and high bitterness (Visser, 1993). One of the most frequently used methods to monitor proteolytic processes on caseins is urea-polyacrylamide gel electrophoresis. On the other hand, tricine-SDS polyacrylamide gel electrophoresis improves the separation, identification and quantification of casein hydrolysates because it allows the visualization of large and small peptides (Pardo & Natalucci, 2001), with the additional advantage of allowing the estimation of molecular masses. Both methods are then suitable to characterize the performance of vegetable rennet in different ways. This preliminary study had the following objectives: the partial characterization of (i) the aspartic proteolytic activity present in flowers of Silybum marianum (L.) Gaertn. (Asteraceae); and (ii) the hydrolytic profile of bovine caseins.

Animals↗

Antimycotic activities of selected plant flora, growing wild in Lebanon, against phytopathogenic fungi.

Petroleum ether (PE) and methanolic extracts of nine wild plant species were tested in vitro for their antimycotic activity against eight phytopathogenic fungi. The efficacy of PE extracts against all pathogens tested was higher than that of methanolic extracts. Wild marjoram (Origanum syriacum) PE extract showed the highest and widest range of activity. It resulted in complete inhibition of mycelial growth of six of eight fungi tested and also gave nearly complete inhibition of spore germination of the six fungi included in the assay, namely, Botrytis cinerea, Alternaria solani, Penicillium sp., Cladosporium sp., Fusarium oxysporum f. sp. melonis, and Verticillium dahlia. The other plant extracts showed differential activities in the spore germination test, but none was highly active against mycelial growth. Inula viscosa and Mentha longifolia were highly effective (>88%) in spore germination tests against five of six fungi tested, whereas Centaurea pallescens, Cichorium intybus, Eryngium creticum, Salvia fruticosa, and Melia azedarach showed >95% inhibition of spore germination in at least two fungi. Foeniculum vulgare showed the least antimycotic activity. Fractionation followed by autobiography on TLC plates using Cladosporium sp. as a test organism showed that O. syriacum PE extracts contained three inhibition zones, and those of Inula viscosa and Cichorium intybus, two, whereas the PE extracts of the remaining plants showed each one inhibition zone. Some of the major compounds present in these inhibition zones were identified by GC-MS. The possibility for using these extracts, or their mixtures, to control plant diseases is discussed.

Alternaria↗

Phytotoxic and antimicrobial activities of catechin derivatives.

(+/-)-Catechin is a potent phytotoxin, with the phytotoxicity due entirely to the (-)-catechin enantiomer. (+)-Catechin, but not the (-)-enantiomer, has antibacterial and antifungal activities. Tetramethoxy, pentaacetoxy, and cyclic derivatives of (+/-)-catechin retained phytotoxicity. The results indicate that antioxidant properties of catechins are not a determining factor for phytotoxicity. A similar conclusion was reached for the antimicrobial properties. Centaurea maculosa (spotted knapweed) exudes (+/-)-catechin from its roots, but the flavanol is not re-absorbed and hence the weed is not affected. The much less polar tetramethoxy derivative may, however, be absorbed and hence be able to cause toxicity. Because of the combination of phytotoxicity and antimicrobial activity, (+/-)-catechin could be a useful natural herbicide and antimicrobial.

Anti-Bacterial Agents↗

Biological monitoring of ozone: the twenty-year Italian experience.

Tropospheric ozone is a growing environmental menace in Italy and in the whole Mediterranean basin. The importance of active biomonitoring of this pollutant with hypersensitive Bel-W3 tobacco plants is stressed, and several examples of field studies carried out in Italy with this technique are presented. Current limitations are discussed, with special emphasis on data quality assessment and the opportunity of adopting easy-to-use kits based on tobacco germlings instead of adult plants. A standardization of methodologies (from cultivation to scoring and data elaboration), also at an international level, is strongly felt to be needed, in order to get official acknowledgement of biomonitoring procedures. Potential educational implications, with the active involvement of students and environmentalists, are shown. Other biological indicators are used, namely sensitive and resistant white clover (Trifolium repens) clones (as descriptors of biomass reduction in crops species) and Centaurea jacea (brown knapweed) as a model species to evaluate the relationship between ozone exposure and effects on the performance and injury symptoms of native plants which are largely used in the framework of European programmes.

Environmental Monitoring↗

Maculosin, a host-specific phytotoxin for spotted knapweed from Alternaria alternata.

Several diketopiperazines have been isolated from liquid cultures of Alternaria alternata, the causal agent of black leaf blight of spotted knapweed, Centaurea maculosa Lam. One of these compounds, maculosin [the diketopiperazine cyclo(-L-Pro-L-Tyr-)], was active in the nicked-leaf bioassay at 10(-5) M; synthetic maculosin possessed chemical and biological activities identical to those of the natural product. Other diketopiperazines isolated from the fungus possessed either less activity or none at all. In tests against 19 plant species, maculosin was phytotoxic only to spotted knapweed. Thus maculosin is a host-specific phytotoxin from a weed pathogen.

Journal Article↗

Novel repeated DNA sequences in safflower (Carthamus tinctorius L.) (Asteraceae): cloning, sequencing, and physical mapping by fluorescence in situ hybridization.

Two novel repetitive DNA sequences, pCtKpnI-1 and pCtKpnI-2, were isolated from Carthamus tinctorius (2n = 2x = 24) and cloned. Both represent tandemly repeated sequences. The pCtKpnI-1 and pCtKpnI-2 clones constitute repeat units of 343-345 bp and 367 bp, respectively, with 63% sequence heterogeneity between the two. Fluorescence in situ hybridization (FISH) was employed on metaphase chromosomes of C. tinctorius using, simultaneously, pCtKpnI-1 and pCtKpnI-2 repeated sequences. The pCtKpnI-1 sequence was found to be exclusively localized at subtelomeric regions on most of the chromosomes. On the other hand, sequence of the pCtKpnI-2 clone was distributed on two nucleolar and one nonnucleolar chromosome pairs. The satellite, and the intervening chromosome segment between the primary and secondary constrictions, in the two nucleolar chromosome pairs were wholly constituted by pCtKpnI-2 repeated sequence. The pCtKpnI-2 repeated sequence, showing partial homology to intergenic spacer (IGS) of 18S-25S ribosomal RNA genes of an Asteraceae taxon (Centaurea stoebe), and the 18S-25S rRNA gene clusters were located at independent, but juxtaposed sites in the nucleolar chromosomes. Variability in the number, size, and location of the two repeated sequences provided identification of most of the chromosomes in the otherwise not too distinctive homologues within the complement. This article reports the start of a molecular cytogenetics program targeting the genome of safflower, a major world oil crop about whose genetics very little is known.

Base Sequence↗

Toxic inhibition of smooth muscle contractility by plant-derived sesquiterpenes caused by their chemically reactive alpha-methylenebutyrolactone functions.

1. Previous studies have shown that extracts of feverfew (Tanacetum parthenium) and parthenolide, a sesquiterpene alpha-methylenebutyrolactone obtained from it, inhibit smooth muscle contractility in a time-dependent, non-specific and irreversible manner. 2. The hypothesis that this toxic effect is due specifically to the presence in the sesquiterpene lactone of the potentially reactive alpha-methylene function was tested on rabbit isolated aortic ring preparations. This was done (a) by comparing the effects of two plant-derived sesquiterpene lactones purified from yellow star thistle (Centaurea solstitialis): cynaropicrin (an alpha-methylenebutyrolactone) and solstitialin 13-acetate (lacking the alpha-methylene function), and (b) by chemically inactivating the alpha-methylene functions in cynaropicrin and parthenolide by reaction with cysteine. 3. The results show that the characteristic smooth muscle inhibitory profile is demonstrated by the two alpha-methylenebutyrolactones (parthenolide and cynaropicrin), but not by the compound lacking this functional group (solstitialin 13-acetate), or by those previously active compounds in which it has been inactivated with cysteine. 4. Thus the alpha-methylene function is critical for this aspect of the toxic pharmacological profile of the sesquiterpene butyrolactones, which are natural products widely distributed in the Compositae family of flowering plants.

Animals↗