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[Mechanism of quercetin as an antidiarrheal agent].

OBJECTIVE: To investigate the antidiarrheal mechanism of quercetin extracted from Psidium guajava L. METHODS: The effects of quercetin on the contractility of guinea pig ileum in vitro and on the peristaltic motion of mouse small intestine in vivo were observed, and the peak value of contractility and peristaltic distance recorded. The inhibitory effect of quercetin on the permeability of the abdominal capillaries was also observed. RESULTS: Quercetin inhibited the contraction of guinea pig ileum in vitro and the peristaltic motion of mouse small intestine, and reduced the permeability of abdominal capillaries. CONCLUSION: Quercetin can inhibit the intestinal movement and reduce capillary permeability in the abdominal cavity, which may be the antidiarrheal mechanism of Psidium guajava L extract.

Animals↗

[The in vitro action of plants on Vibrio cholerae].

Natural products of several plants, according to the geographic location, are used by Peruvian people in the popular treatment of diarrhea, with good success. When cholerae cases appeared in Peru, we were interested to know the "in vitro" effect against Vibrio cholerae 01, of these useful plants to treat diarrhea. The following plants were tested: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Punica granatum, Malus sativa, Cydonia oblonga, Chenopodium ambrosoides, Krameria triandria, Tea chinensis, Daucus carota, Persea gratissima, Psidium guayaba and Lippia dulcis. Decoction or infusion of the plants were used in the "in vitro" experiments. The following plants showed no "in vitro" effect against V. cholerae: Cichorium intybus, Althaea officinalis, Psorela glandulosa, Geranium maculatum, Chenopodium ambrosoides, Krameria triandria, Psidium guayaba, Lippia dulcis and Daucus carota. Decoction of Malus sativa and Cydenia oblonga showed bactericidal effect for their acidity and stone avocado (Persea gratissima) a late bactericidal effect. Tea infusión and the decoction of Punica granatum peel, showed the best bactericidal effect and we suggest to use them as to stop cholera spreading.

Hydrogen-Ion Concentration↗

Inhibitory effect of Thai plant extracts on P-glycoprotein mediated efflux.

Curcuminoids from Curcuma longa L. and extracts of Psidium guajava L., Andrographis paniculata (Burm. f.) Nees, Phyllanthus emblica L. and Solanum trilobatum L. were investigated for their inhibitory effect on P-glycoprotein (P-gp) on the efflux transport of rhodamine 123 (Rho-123 ) in Caco-2 cells and rat ileum. Of the five tested samples, curcuminoids and an extract of P. guajava showed the highest inhibitory effect on P-gp mediated efflux of Rho-123 in Caco-2 cells. Additionally, they were found to have equal potential in inhibiting Rho-123 efflux transport from serosal to mucosal surfaces of the rat ileum.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The "heterolytic hydroperoxide lyase" is an isomerase producing a short-lived fatty acid hemiacetal.

To elucidate the reaction mechanism of hydroperoxide lyase (HPL), the enzyme from guava (Psidium guajava) fruits, was incubated for 10-60 s at 0 degrees C with 13-HPOT. The products were rapidly extracted and derivatized by trimethylsilylation. Two trapping products, namely the trimethylsilyl ether/ester derivatives of the hemiacetal 12-(1'-hydroxy-3'-hexenyloxy)-9,11-dodecadienoic acid and the enol (9Z,11E)-12-hydroxy-9,11-dodecadienoic acid, were detected by gas chromatography-mass spectrometry (GC-MS) analyses. The structural assignments were supported by mass spectra recorded for (a) hydrogenated products; (b) products biosynthesized from [9,10,12,13,15,16] 13-HPOT or [(18)O(2)]13-HPOT; (c) chemically prepared reference compounds. Kinetic experiments showed that the hemiacetal and enol were both unstable and transiently appearing compounds (half-lives, ca. 20 s and 2 min, respectively). Hemiacetal and enol biosynthesized from [(18)O(2)]13-HPOT retained two and one (18)O atoms, respectively, whereas no (18)O was incorporated from [(18)O]water. The data demonstrated that: (1) the true enzymatic product formed from 13-HPOT in the presence of HPL is a short-lived hemiacetal; (2) the hemiacetal spontaneously dissociates into (3Z)-hexenal and the unstable enol form of (9Z)-12-oxo-9-dodecenoic acid; (3) the enzymatic isomerization of 13-HPOT into the hemiacetal occurs homolytically.

Acetals↗

Hydroperoxide lyases (CYP74C and CYP74B) catalyze the homolytic isomerization of fatty acid hydroperoxides into hemiacetals.

The conversion of linoleic acid 9-hydroperoxide (9-HPOD) by recombinant melon (Cucumis melo L.) hydroperoxide lyase (HPL, CYP74C subfamily) was studied. Short (5 s-1 min) incubations at 0 degrees C followed by rapid extraction and trimethylsilylation made it possible to trap a new unstable (t(1/2) <30 s) product, i.e. the hemiacetal (1'E,3'Z)-9-hydroxy-9-(1',3'-nonadienyloxy)-nonanoic acid. Identification was performed by GC-MS analysis and substantiated by the formation of trimethylsilyl 9-trimethylsilyloxy-9-nonyloxy-nonanoate upon catalytic hydrogenation and by (2)H-labelling experiments. Both (18)O atoms of [(18)O(2)-hydroperoxy]9-HPOD were incorporated into the hemiacetal. Along with the hemiacetal, three chain-cleavage products, i.e. the enol (1E,3Z)-nonadienol and the hydrates of 3(Z)-nonenal and 9-oxononanoic acid, were trapped as their trimethylsilyl derivatives. The kinetics of (18)O incorporation from [(18)O(2)]9-HPOD provided strong evidence that the cleavage products originated in the hemiacetal. Linolenic and linoleic acid 13-hydroperoxides served as substrates for recombinant HPLs of melon, alfalfa (Medicago sativa) and guava (Psidium guajava), and in each case hemiacetals and enols were detectable by the trapping technique. The data obtained demonstrated that CYP74C and CYP74B HPLs act as isomerases performing a homolytic rearrangement of fatty acid hydroperoxides into short-lived hemiacetals which upon decomposition produce 3(Z)-nonenal, 3(Z)-hexenal and other short chain aldehydes.

Acetals↗

Anti-proliferative activity of essential oil extracted from Thai medicinal plants on KB and P388 cell lines.

Anti-proliferative activity of essential oil from 17 Thai medicinal plants on human mouth epidermal carcinoma (KB) and murine leukemia (P388) cell lines using MTT assay were investigated. An amount of 1 x 10(4)cells/well of KB cell line and 1 x 10(5) cells/well of P388 cell line were treated with the oil samples at different concentrations ranging from 0.019 to 4.962 mg/ml. In KB cell line, Guava (Psidium guajava L.) leaf oil showed the highest anti-proliferative activity with the IC(50) value of 0.0379 mg/ml (4.37 times more potent than vincristine) whereas Sweet Basil (Ocimum basilicum L.) oil gave the highest anti-proliferative activity with the IC(50) value of 0.0362 mg/ml (12.7 times less potent than 5-FU) in P388 cell line. The results demonstrated the potential of essential oil from Thai medicinal plants for cancer treatment.

Animals↗

Atmospheric dry deposition to leaf surfaces at a rural site of India.

Dry deposition flux of major ions (Na+, K+, Ca2+, Mg2+, NH4+, F-, Cl-, NO3- and SO4(2-) to natural surfaces [guava (Psidium guyava) and peepal (Ficus religiosa) leaves] are determined at Rampur, a rural site of semi-arid region of India. Dry deposition flux is the highest for Ca2+ on guava leaves and for NH4+ on peepal leaves. Overall dry deposition flux is higher on guava leaves than of peepal leaves. The variation in deposition flux may be due to surface characteristics (surface roughness) and arrangement of leaves. Peepal leaves are arranged along the axis of the stem, whereas guava leaves are at right angles to the stem. The deposition flux of cations contributes 66% and 76% of dry deposition of all major ions on guava and peepal leaves, respectively as soil is major contributor towards dry deposition flux in tropical regions. ANOVA revealed no significant seasonal difference in deposition, although there is a trend for higher in winter. Deposition velocities of NH4+, NO3- and SO4(2-) are greater on guava leaves than peepal leaves, which can be attributed to the rougher surface of the guava leaf.

Air Pollutants↗

Possible modulating actions of plant extracts on the chromosome breaking activity of MMC and Ara-C in human lymphocytes in vitro.

Plants popularly used as medicine have been seen as promising natural agents by the pharmaceutical industry. In the present study the action of Psidium guajava L. (Pg) and Achillea millefolium L. (Am) infusions on chromosomal aberration formation in human lymphocyte system in vitro was assessed, associating them with the alkylating agent mitomycin C (MMC) and the DNA repair inhibitor cytosine-beta-arabin-furanoside (Ara-C). The cells were cultivated for 72 h and treated continuously with Pg and the Am infusions at dosages of 2.62 x 10(-4) g and 3.5 x 10(-4) g/ml culture medium, respectively. Treatments with MMC (0.30 microg/ml) or Ara-C (5 x 10(-7) microg/ml) were administered after 48 h of cell culture. Each samples (five individual) were exposed to nine treatments (control with PBS; Pg; Am; MMC; MMC+Pg; MMC+Am; Ara-C; Ara-C+Pg; and Ara-C+Am) and 100 cells were analyzed per cell culture. The used doses of each infusion did not cause clastogenic effects significantly different to the negative control (control=1%; Pg=2.2%; Am=1.8%). Nevertheless, the aberrant cell frequency after MMC treatment was significantly increased by the Am infusion (MMC=32.4%; MMC+Pg=36.2%; MMC+Am=44%), especially when the chromatid break types number was scored (MMC=151; MMC+Pg=173; MMC+Am=249). Regarding DNA repair inhibition by Ara-C, the Pg infusion caused a significant reduction in aberrant cell frequency (Ara-C=15.8%; Ara-C+Pg=11%; Ara-C+Am=14.4%), only when the chromatid break types number was scored (Ara-C=63; Ara-C+Pg=40; Ara-C+Am=58). These results indicate that the plant infusions per se do not have clastogenic activity, but can influence the clastogenic action of MMC and Ara-C on DNA break induction, in vitro.

Achillea↗

Detection of an enol intermediate in the hydroperoxide lyase chain cleavage reaction.

Guava (Psidium guajava) hydroperoxide lyase (HPL) preparations were incubated with [1-(14)C](9Z,11E,13S,15Z)-13-hydroperoxy-9,11,15-octadecatrienoic acid for 1 min at 0 degrees C, followed by rapid extraction/trimethylsilylation. Analysis of the trimethylsilylated products by gas chromatography-mass spectrometry and radio-high-performance liquid chromatography revealed a single predominant (14)C-labelled compound, identified by its (1)H-nuclear magnetic resonance, ultraviolet and mass spectra as the trimethylsilyl ether/ester of (9Z,11E)-12-hydroxy-9,11-dodecadienoic acid. Longer time incubations afford smaller yield of this enol due to its partial tautomerization into (9Z)-12-oxo-9-dodecenoic acid. The data obtained demonstrate that formation of (9Z)-12-oxo-9-dodecenoic acid in the HPL reaction is preceded by unstable enol oxylipin, and further suggest that hemiacetals are the true products of HPL catalysis.

Aldehyde-Lyases↗

Simple detection method of powerful antiradical compounds in the raw extract of plants and its application for the identification of antiradical plant constituents.

A simple detection method for a powerful radical scavenging compound in a mixture containing a large variety of compounds, such as the raw extract of edible plants, was developed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) as the radical reagent. The method was established on the basis of the features of the typical chain-breaking antioxidation reaction mechanism, which suggests that the radical scavenging antioxidant should be converted to other stable nonradical compounds during the reaction. This method requires only a simple HPLC instrument, and the disappearance or decrease in the peak intensity, which is induced by the addition of DPPH. This change is monitored by the HPLC to detect the powerful radical scavenger from the complex mixture. The method was applied to the detection and identification of the most powerful antiradical compound in the extracts of three antioxidatively active plant extracts (Psidium guajava, Citrus depressa, and Hypericum chinense). The radical scavenging efficiency of a newly identified compound from H. chinense was also compared with that of Trolox and catechin using the method.

Antioxidants↗

Contribution of natural food sources to reproductive behaviour, fecundity and longevity of Ceratitis cosyra, C. fasciventris and C. capitata (Diptera: Tephritidae).

The influence of food sources comprising the natural diet on the reproductive behaviour, fecundity and longevity of three African fruit flies Ceratitis cosyra (Walker), C. fasciventris (Bezzi) and C. capitata (Wiedemann) was investigated. Three natural food sources, varying in protein and sugar content, were evaluated. These included bird droppings (farm chicken), aphid honeydew and guava (Psidium guajava L.) juice. For C. fasciventris and C. capitata, flies fed on a protein-rich diet displayed higher frequency of calling, mating and oviposition than flies fed on a protein-poor diet, whilst for C. cosyra, quality of diet significantly influenced the mating behaviour of the flies, but not the calling and oviposition behaviour. Net fecundity rates were lowest for C. fasciventris and C. capitata when fed only on guava juice (0.1, 2.6 eggs per female, respectively), and higher for those on a diet of honeydew only (9.5, 33.8 eggs per female, respectively) and a combined diet of guava, honeydew and chicken faeces (11.8, 25.8 eggs per female, respectively). For C. cosyra, due to low numbers of eggs collected, no significant differences in fecundity between diets could be detected. All species fed only on a diet of chicken faeces since emergence died within the first three days of adult life without laying eggs, but when carbohydrates were provided by addition of guava juice and honeydew, the longevity of the flies was sustained for more than four weeks after adult emergence. The practical implications of these findings for control purposes are discussed.

Analysis of Variance↗

A stochastic flowering model describing an asynchronically flowering set of trees.

A general stochastic model is presented that simulates the time course of flowering of individual trees and populations, integrating the synchronization of flowering both between and within trees. Making some hypotheses, a simplified expression of the model, called the 'shoot' model, is proposed, in which the synchronization of flowering both between and within trees is characterized by specific parameters. Two derived models, the 'tree' model and the 'population' model, are presented. They neglect the asynchrony of flowering, respectively, within trees, and between and within trees. Models were fitted and tested using data on flowering of Psidium cattleianum observed at study sites at elevations of 200, 520 and 890 m in Reunion Island. The 'shoot' model fitted the data best and reproduced the strong irregularities in flowering shown by empirical data. The asynchrony of flowering in P. cattleianum was more pronounced within than between trees. Simulations showed that various flowering patterns can be reproduced by the 'shoot' model. The use of different levels of organization of the general model is discussed.

Computer Simulation↗

Guaijaverin -- a plant flavonoid as potential antiplaque agent against Streptococcus mutans.

AIMS: The aim of the present study was to investigate the anti-Streptococcus mutans activity and the in vitro effects of subminimal inhibitory concentrations of guaijaverin isolated from Psidium guajava Linn. on cariogenic properties of Strep. mutans. METHODS AND RESULTS: Bioautography-directed chromatographic fractionation, yield biologically active compound, quercetin-3-O-alpha-l-arabinopyranoside (guaijaverin), from crude methanol extract of P. guajava. Growth-inhibitory activity of the compound against Strep. mutans of both clinical and type strain cultures was evaluated. The anti-Strep. mutans activity of the guaijaverin was found to be bacteriostatic, both heat and acid stable and alkali labile with the minimum inhibitory concentration (MIC) of 4 mg ml(-1) for MTCC 1943 and 2 mg ml(-1) for CLSM 001. The sub-MIC concentrations (0.0078-2 mg ml(-1)) of the guaijaverin were evaluated for its cariogenic properties such as acid production, cell-surface hydrophobicity, sucrose-dependent adherence to glass surface and sucrose-induced aggregation of Strep. mutans. CONCLUSIONS: The active flavonoid compound, quercetin-3-O-alpha-l-arabinopyranoside (guaijaverin) demonstrated high potential antiplaque agent by inhibiting the growth of the Strep. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the new growth-inhibitory compound guaijaverin against Strep. mutans and led to the acceptance of traditional medicine and natural products as an alternative form of health care.

Bacterial Adhesion↗

Identification of flavonoids and their glycosides by high-performance liquid chromatography with electrospray ionization mass spectrometry and with diode array ultraviolet detection.

Identification of flavonoids and flavonoid glycosides was carried out on Psidium guajava Linn leaves by means of high-performance liquid chromatography ultraviolet (HPLC-UV) analysis and HPLC mass spectrometry. By using HPLC-UV, two known phenolics (gallic acid and quercetin) and five newly reported ones (procatechuic acid, chlorogenic acid, caffeic acid, kaempferol and ferulic acid) were identified in alcohol guava leaf extract. Structural information about the compounds was obtained from the retention times, the UV spectra and mass spectra without the need to isolate the individual compounds. Two flavonoids (quercetin and kaempferol) and four flavonoid glycosides (three known components, quercetin 3-O-alpha-L-arabinoside, quercetin 3-O-beta-D-glucoside and quercetin 3-O-beta-D-galactoside, along with one novel compound, kaempferol-glycoside) and three other unknown compounds have been identified in the fractions.

Chromatography, High Pressure Liquid↗

Variability in habitat, diet, and social structure of Hapalemur griseus in Ranomafana National Park, Madagascar.

This study investigated the effects of habitat on the diet, population density, and social structure of a small-bodied folivore, Hapalemur griseus. Three groups of H. griseus were followed at two study sites (Tala and Vato) within Ranomafana National Park in southeastern Madagascar. These two sites differed in degree of habitat disturbance, forest composition, and forest structure, as determined by botanical plots (50 x 10 m) which were monitored twice monthly in the home ranges of three study groups (group T, n = 4; group V1, n = 4; group V2, n = 2). Tala has experienced more habitat disturbance compared to Vato, and this appeared to influence population density and group size. The H. griseus that inhabit Tala occur at much higher densities and in larger social groups than the H. griseus at Vato. These results may be attributable to: 1) a greater number of potential food sources, 2) a more dense resource base, and/or 3) higher-quality resources at Tala for this folivore. In general, dietary diversity for all groups was low, and groups exploited similar plant species. However, within this small range of food sources, each study group specialized in food sources found in the highest abundance within their home range. This resulted in dietary differences between groups within sites, as well as differences between sites. Introduced tree species such as Chinese guava (Psidium cattleyanum) may have greatly impacted the diet and social structure of those groups at Tala, whose home ranges included this food source. In contrast, Vato group 1 ingested more new and mature leaves of Ficus spp., and Vato group 2 primarily ate spider bamboo (Nastus elongatus). In conclusion, it seems that microhabitat differences, which may be related to habitat disturbance and/or other factors such as topography, influenced the food species ingested by H. griseus. Overall habitat quality, which is likely also affected by habitat disturbance, influenced general diet, population density, and group size.

Animals↗

Effects of some natural extracts on the acetylcholine release at the mouse neuromuscular junction.

Natural extracts have been proved to be useful in different human pathological conditions. The scientific consideration of the therapeutic potential of plant extracts is still inappropriate due to the lack of both pharmacological and epidemiological basic studies. Here, we started from an electrophysiological point of view, a study on the effects of two extracts on the acetylcholine (ACh) release at the neuromuscular junction. The extracts purified from Sugar cane (policosanol) and Psidium guajava (quercetin) have been submitted to this study. The wide epidemiology of these agents suggests therapeutic potentials not yet well outlined at the basic level. Our data demonstrate some interactions in the modulation of the ACh release at the mouse neuro-muscular junction, which are well correlated with the suggested molecular mechanisms. Policosanol enhances to a small extent either the spontaneous or the evoked ACh release. Furthermore, an increase of the rate of the conformational change induced at the nicotinic receptor-channel complex by ACh is also observed. Quercetin induced a reduction of the ACh evoked release. The possibility that this effect could be ascribed to some interaction with presynaptic calcium channel is noteworthy. The results are discussed in terms of a possible interference with acetylcholinesterase by policosanol and of a presynaptic molecular action of quercetin modulating the cytosolic calcium concentration.

Acetylcholine↗

The toxicity of chlorothalonil to aquatic fauna and the impact of its operational use on a pond ecosystem.

Chlorothalonil is a fungicide whose heavy use in eastern Canada gives it the potential for significant aquatic contamination. Laboratory bioassays and field treatments of a pond system were undertaken to determine the toxic effects of chlorothalonil on aquatic fauna. The 96-h LC50 of technical chlorothalonil for rainbow trout (Oncorhynchus mykiss) was 76 micrograms/L and was not significantly different (p less than 0.05) from that of the formulated product (Bravo 500). The 96-h LC50 of Bravo 500 for blue mussels (Mytilus edulis) and clams (Mya arenaria) was 5.9 mg/L and 35.0 mg/L respectively, while its 48-h LC50 to Daphnia magna was between 130 micrograms/L and 200 micrograms/L. Chlorothalonil exposure of Daphnia to concentrations as low as 32 micrograms/L significantly (p less than 0.05) increased the time to production of first young, but there were no delayed effects on number of young produced or growth at concentrations of 180 micrograms/L or less. Chlorothalonil was initially accumulated by blue mussels to concentrations approximately ten times greater than exposure concentrations; however, tissue concentrations returned to the same level as exposure concentrations within 96 h. Spraying of ponds resulted in mortality of caged water boatmen (Sigara alternata) and threespine stickleback (Gasterosteus aculeatus) which could be related to chlorothalonil exposure, however, caddisfly larvae (Limnephilus sp.), freshwater clams (Psidium sp.), water beetles (Haliplus sp.), scud (Gamarus spp.) and midge larvae (Chironomidae) did not suffer substantial chlorothalonil-induced mortality. Changes in endemic benthic invertebrate abundance after sprays were not remarkable or related to treatment. Faunal impacts in the pond were generally of a smaller magnitude than were predicted by bioassay results.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Evaluation of air pollution phytotoxicity downwind of a phosphate fertilizer factory in India.

The effects of air pollution on plants downwind of a fertilizer factory at Udaipur, India, were studied using three woody perennials. Seedlings of these species including a shrub (Carissa carandas L.), a leguminous avenue tree (Cassia fistula L.) and a fruit tree (Psidium guajava L.) were grown in earthen pots at different study sites receiving varying levels of air pollution input. Changes in plant growth, morphological characteristics, photosynthetic pigment, ascorbic acid, N and S contents and in dry matter allocation were considered in relation to the status of ambient air quality. Observations with these parameters have indicated that the ambient air around the factory contained pollutants at phytotoxic levels. Plant height, basal diameter, conopy area, leaf area and chlorophyll, ascorbic acid and foliar-N concentrations decreased with increasing pollution load. However, foliar-S increased slightly at polluted sites. Air pollution load around the factory have also altered the biomass allocation. Root:shoot ratios increased in C. fistula and P. guajava at polluted sites. In contrast, for C. carandas the above ground parts, where foliage assumed predominance showed precedence over the root growth. This species responded characteristically to air pollution stress by allocating more of its photosynthate towards leaf production and shoot growth.

Air Pollutants↗