Paralyzing and myotoxic effects of a recombinant bothropstoxin-1 (BthTX-I) on mouse neuromuscular preparations

As a first step to investigate the structure-function relationship of bothropstoxin-1 (BthTX-1), a myotoxin from Bothrops jararacussu snake venom, Our group previously cloned a recombinant toxin (rBthTX-1) in Escherichia coli. The aim or this work was to characterize the biological activities of this rBthTX-1 (1.0 mu M) in both phrenic-diaphragm and extensor digitorum longus preparations in vitro, by means of myographic and morphologic techniques. Native BthTX-1 (1.0 mu M) was used as a standard. The influence of heparin (27.5 mu g/ml) upon the biological activities of both toxins was also investigated. rBthTX-1 had similar effects to the native toxin inducing blockage of both directly and indirectly evoked contractions in phrenic-diaphragm preparations, and muscle damage characterized by edema, round fibers, and cell areas devoid of myofibrils. Interestingly the paralyzing activity of rBthTX-1 was slightly more potent than the native toxin. Heparin prevented paralyzing and myotoxic effects of both the native and recombinant toxins. This work shows that rBthTX-1 was expressed in a fully active form, and presents a biological profile similar to the native toxin. (c) 2005 Elsevier GmbH All rights reserved.

Antiulcerogenic and analgesic effects of the Austroplenckia populnea extracts in mice

Austroplenckia populnea (Reiss.) Lund. is a Brazilian Cerrado plant belonging to the Celastraceae family. Hexane and methanol extracts of leaves were investigated for their antiulcerogenic (ethanol and indomethacin/bethanecol induced gastric damage) and analgesic (writhing and tail-flick tests) activities in mice. Acute toxic effects also were evaluated. Oral administration of both extracts at a dose of 1000 mg/kg significantly reduced the total area of the lesion, the relative area of lesion and the ulcerative index in ethanol-induced gastric damage, but both extracts were inactive in the indomethacin/bethanecol-induced gastric damage test. A dose dependent effect was determined with the hexane extract in the ethanol-induced lesions test. The hexane and methanol extracts reduced the number of contortions in the writhing test, but both extracts were inactive in the tail-flick immersion test. Copyright (C) 2002 John Wiley Sons, Ltd.

Antiulcerogenic and analgesic effects of Maytenus aquifolium, Sorocea bomplandii and Zolernia ilicifolia

Maytenus aquifolium (Celastraceae), Sorocea bomplandii (Moraceae) and Zolernia ilicifolia (Fabaceae) are native plants from the Tropical Atlantic Forest (Mata Atlantica, Brazil) known a 'espinheira-santa'. These plants are traditionally used as analgesic and antiulcerogenic medicine, with the same traditional uses of the true 'espinheira-santa' (Maytenus ilicifolia, Celastraceae), an efficient antiulcerogenic agent. Pharmacological and toxicological studies with these plants have not been carried out. The purpose in this study was to evaluate the efficacy (analgesic and antiulcerogenic activities), safety (acute toxicity) and quality (phytochemical profile) of these three plants. The analgesic effect was analyzed by writhing and tail flick tests, while anti Ulcerogenic effect was performed through ulcer induction by ethanol and indomethacin/bethanecol assays. LD., and acute toxic effects, as well as phytochemical profiles of all plants also were carried. Surprisingly, the three plants showed analgesic and antiulcerogenic effects at dose of 1000 mg/kg, v.o. Maytenus aquifolium lowering all ulcerogenic parameters (ethanol test), but increased the ulcerogenic effects in the indoniethacin/bethanecol test. Sorocea bomplandii produced antiulcerogenic effects in both experimental models used, while Zolernia ilicifolia showed significant effects only in indomethacin/bethanecol-induced gastric lesions. Pre-treatment with Zolernia ilicifolia induced someone toxic effects, A phytochemical profile for each plant species was determined and its main chemical classes of compounds were described. (C) 2001 Elsevier B.V. Ireland Ltd. All rights reserved.

Hymenaea stigonocarpa Mart. ex Hayne: A Brazilian medicinal plant with gastric and duodenal anti-ulcer and antidiarrheal effects in experimental rodent models

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Byrsonima intermedia A. Juss.: Gastric and duodenal anti-ulcer, antimicrobial and antidiarrheal effects in experimental rodent models

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electrical and mechanical effects of hyoscine butylbromide on the human stomach: a non-invasive approach

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Hypoglycaemic and antioxidant effects of onion, Allium cepa: dietary onion addition, antioxidant activity and hypoglycaemic effects on diabetic rats

The purpose of the present study was to discover the relative potency of onion, Allium cepa, with respect to its hypoglycaemic and hypolipidaemic effects on the diabetic situation, and the association of these effects with the potential against oxidative stress. Male Wistar rats were divided into four groups. A normal control ( group A), and a non-diabetic group ( group B) were treated daily with 1 ml A. cepa solution (0.4 g A. cepa/rat). Groups C and D were made diabetic by an intraperitoneal injection of streptozotocin (STZ) ( 60 mg/kg body weight) in citrate buffer ( pH 6.3). These animals ( groups C and D) were the STZ diabetic control and STZ diabetic rats with onion intake, respectively. Onion increased the fasting serum high-density lipoprotein levels, and demonstrated alleviation of hyperglycaemia in STZ diabetic rats. The hypoglycaemic and hypolipidaemic actions of A. cepa were associated with antioxidant activity, since onion decreased superoxide dismutase activities while no increased lipid hydroperoxide and lipoperoxide concentrations were observed in diabetic rats treated with A. cepa.

Gastroprotective and antisecretory effects of Ailanthus excelsa (Roxb)

Ailanthus excelsa (Roxb), an Egyptian medicinal species highly important for treating numerous diseases, was investigated against experimentally induced gastric ulcer in rodents. We evaluated the gastroprotective effect of four extracts (petroleum ether, diethyl ether, chloroform, and methanol) of A. excelsa bark by using the ethanol-induced gastric lesion model. The pretreatment of animals with methanolic, petroleum ether, and chloroformic extracts (100 mg/kg, oral (p.o.)) from A. excelsa significantly reduced gastric lesion induced by ulcerogenic agent (56, 47, and 70%, respectively) when compared with animals pretreated with vehicle. However, the diethyl ether pretreatment led to the least gastric lesion damage (83%), similar to the standard antiulcer drug, cimetidine, at the same dose (100 mg/kg, p.o.). The lower effective dose of diethyl ether extract, as well as cimetidine, given by intraduodenal route, significantly increased the pH values and reduced the acid output of gastric juice. Sterols, triterpenes, and quassinoids are present in the diethyl ether extract of A. excelsa stem bark, which presented the best gastroprotective action among the studied extracts. Our study confirmed the traditional indications of A. excelsa for the treatment of gastric ulcer.

Correlation between antigenemia of Paracoccidioides brasiliensis and inhibiting effects of plasma in patients with paracoccidioidomycosis

Metabolites produced by pathogenic fungi may be involved in the pathogenesis of fungal infections consequently altering the defence mechanisms of the host. In this study the levels of Paracoccidioides brasiliensis antigens detected in the plasma of patients with paracoccidioidomycosis correlated with the suppression index detected by the low mitogenic response of peripheral blood mononuclear cells (PBMC) to phytohaemaglutinin (PHA). This inhibitory effect on lymphoproliferation was observed in the plasma of 58% of the patients, suggesting the presence of inhibitory factors. Plasma samples from paracoccidioidomycosis patients having or not having inhibitory factors showed no significant effect on chromosomes of lymphocytes from healthy individuals, However, these plasmas had a suppressive activity on the blastogenic response of these lymphocytes stimulated with PHA, that was independent of a cytotoxic effect. P. brasiliensis antigens added to the proliferative response of PBMC from healthy individuals stimulated or not stimulated with PHA showed a dose-dependent suppressor effect, reproducing the inhibitory effect of patients' plasma. We suggest that the antigens of P, brasiliensis present in the plasma of patients, even at low concentrations, can play an important role in the reduction of the cellular immune response and in the genesis of the immuneregulatory disturbances observed in paracoccidioidomycosis.

Ischaemia and reperfusion effects on skeletal muscle tissue: morphological and histochemical studies

This was a study on the oxidative stress due to ischaemia (I) and reperfusion (R) in skeletal muscle tissue. Using a tourniquet, groups of rats were submitted to ischaemia for 4 h, followed by different reperfusion periods. The animals were divided in four groups: control; 4 h of ischaemia (IR); 4 h of ischaemia plus 1 h reperfusion (IR-1 h); 4 h of ischaemia plus 24 h reperfusion (IR-24 h); and 4 h of ischaemia plus 72 h reperfusion (IR-72 h). At the end of the procedures, samples of soleus muscle were collected and frozen in n-hexane at -70 degrees C. Cryostat sections were submitted to haematoxylin-eosin, succinate dehydrogenase (SDH) and nicotinamide adenine dinucleotide-tetrazolium reductase (NADH-TR) stains. An additional muscle sample was processed for electron microscopy. No alterations were found in control animals. IR group showed fibres had normal aspect besides some round, acidophilic and hypertrophic fibres. There were several fibres with angular outlines and smaller diameters in this group compared with control group. NADH-TR/SDH reaction was moderately intense in most fibres. In some fibres, cytoplasm showed areas without activity and other fibres had very intense reactivity. IR-1 h group showed oedema hypercontracted fibres with disorganized myofibrils, mitochondria with focal lesions and dilated sarcoplasmic reticulum. NADH-TR/SDH reaction was moderate to weak. IR-24 h showed intense inflammatory infiltrate in the endomysium and perimysium. NADH-TR/SDH reaction was similar to IR-1 h. IR-72 h showed necrotic fibres, areas with inflammatory infiltrate, reduced muscle fibres at different stages of necrosis and phagocytosis, and many small round and basophilic fibres characterizing a regeneration process. NADH-TR/SDH reaction was weak to negative. Our results suggest that ischaemia and the subsequent 1-, 24- and 72-h reperfusions induced progressive histological damage. Although progressive, it may be reversible because there were ultrastructural signs of recovery after 72-h reperfusion. This recovery could in part be due to the low oxidative stress identified by the morphological and histochemical analysis.

N-Acetylcysteine an Allium Plant Compound Improves High-Sucrose Diet-Induced Obesity and Related Effects

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Independent and interactive effects of a top and an intermediate fish species on the food web structure of a tropical stream

Two fish species, one top predator (Imparfinis mirini) and one intermediate detritivorous species (Hisonotus depressicauda), were experimentally manipulated to evaluate their relative importance in structuring the periphytic community, as well as their effects on the other trophic levels. An enclosure experiment was conducted in the Potreirinho creek, a second order tributary of Paranapanema River, SE Brazil. Five treatments were used: enclosure of the predator species. enclosure of the detritivorous species, enclosure of both together, exclusion of all fish species (closed control cage), and cage open to all fish community, (open control). Through direct and indirect effects, I. mirini, when alone gave rise to a trophic cascade that resulted in a positive effect on algal resources. Through direct effects, H. depressicauda. when alone, reduced the amount of organic matter, resulting in a positive indirect effect on algae. In addition, when the two species were enclosed together, only the effects determined by the detritivorous species were present. The results indicate the important role of the intermediate detritivorous species in the maintenance of the composition and trophic structure of the analyzed community by reducing the effects caused by the top predator.

Genotoxic, mutagenic and cytotoxic effects of the commercial dye CI Disperse Blue 291 in the human hepatic cell line HepG2

Textile dyes are discarded into the aquatic ecosystem via industrial effluents and potentially expose humans and local biota to adverse effects. The commercial dye CI Disperse Blue 291 which contains the aminoazobenzene 2-[(2-bromo-4,6-dinitrophenyl)azo]-5(diethylamino)-4-methoxyacetanilide (CAS registry no. 56548-64-2), was tested for genotoxicity and cytotoxicity in the human hepatoma cell line HepG2, using the comet assay, micronucleus (MN) test and a cell viability test. Five different concentrations of the test compound were examined: 200 mu g/ml, 400 mu g/ml, 600 mu g/ml, 800 mu g/ml and 1000 mu g/ml. An increase in comet tail length and in the frequency of MN was detected with exposure of cells to concentrations of the commercial dye from 400 pg/ml. Furthermore, the dye was found to decrease cell viability. The results of this study demonstrate for the first time the genotoxic and mutagenic effects of the dye CI Disperse Blue 291 in mammalian cells, thus stressing the need to develop non-mutagenic dyes and to invest in improving the treatment of effluents. These measures will help to prevent harmful effects that these compounds can have on humans and aquatic organisms that come in contact with them. (C) 2007 Elsevier Ltd. All rights reserved.

Forest fragment size and microhabitat effects on palm seed predation

The establishment of plant species depends crucially on where the seeds are deposited. However, since most studies have been conducted in continuous forests, not much is known about the effects of forest fragmentation on the maintenance of abiotic and biotic characteristics in microhabitats and their effects on seed survival. in this study, we evaluated the effects of forest fragmentation on the predation upon the seeds of the palm Syagrus romanzoffiana in three microhabitats (interior forest, forest edge and gaps) in eight fragments of semi-deciduous Atlantic forest ranging in size from 9.5 ha to 33,845 ha in southeastern Brazil. Specifically, we examined the influence of the microhabitat structure, fauna and fragment size on the pattern of seed predation. Fragments < 100 ha showed similar abiotic and biotic characteristics to those of the forest edge, with no seed predation in these areas. Forest fragments 230-380 ha in size did not present safe sites for S. romanzoffiana seed survival and showed high seed predation intensity in all microhabitats evaluated. In fragments larger than 1000 ha, the seed predation was lower, with abiotic and biotic differences among gaps, interior forests and forest edges. In these fragments, the survival of S. romanzoffiana seeds was related to squirrel abundance and interior forest maintenance. Based on these results, we concluded that there are no safe sites for S. romanzoffiana seed establishment in medium- and small-sized fragments as result of the biotic and abiotic pressure, respectively We suggest that on these forest fragments, management plans are needed for the establishment of S. romanzoffiana, such as interior forest improvement and development in small-sized sites in order to minimize the edge effects, and on medium-sized fragments, we suggest post-dispersal seed protection in order to avoid seed predation by vertebrates. our findings also stress the importance of assessing the influence of forest fragmentation on angiosperm reproductive biology as part of the effective planning for the management of fragmented areas. (c) 2006 Elsevier Ltd. All rights reserved.

Genotoxic and mutagenic effects of fipronil on mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)