Toxicologic evaluation of acute and subacute oral administration of Cucurbita maxima seed extracts to rats and swine

The extract prepared from dried seeds of Cucurbita maxima was administered to rats and pigs. Following a single dose or 4 weeks of daily oral administration, the extract produced no changes in serum glucose, urea, creatinine, total protein, uric acid, GOT, GPT, LDH or blood counts. Urine analysis (urea, uric acid, creatinine, total protein, Na and K), as well as histopathological investigation, showed no abnormalities. These results taken as a whole indicate that the seeds of C. maxima as used in Brazilian folk medicine are not toxic for rats and swine.

Cutaneous hypersensitivity induced in dogs and guinea-pigs by extracts of the tick Rhipicephalus sanguineus (Acari: Ixodidae)

The cutaneous hypersensitivity induced by Rhipicephalus sanguineus tick extract in dogs (natural host) and guinea-pigs (laboratory host) was evaluated. The left ear of infested and control (tick-bite naive) dogs and guinea-pigs was injected intradermally with an extract from unfed adult ticks and the right ear with phosphate buffered saline (PBS). Ear thickness variations were then measured after 10 min and 1, 2, 6, 18, 24, 48, 72 and 96 h post-injection. Results were expressed as percentual changes in the ear thickness in relation to pre-inoculation values. The final variation in ear thickness induced by the extract was given by subtracting, in each animal, the right ear percentual increase from that of the left ear. Guinea-pigs were tested at two different times following infestation and with two different doses of extract. Infested guinea-pigs from the three experiments developed an immediate (within the first 2 h post-inoculation) and a strong delayed reaction (24 h) to the extract. Dogs, unlike guinea-pigs, developed only a strong immediate reaction whereby an 80% increase in ear thickness was observed. Control animals, with the exception of one dog, did not develop any significant reaction to the extract. Only mild reactions were induced by PBS in the right ear of all animals. The correlation between the absence of a strong delayed type reaction to tick extract and the lack of resistance of the natural host to R. sanguineus tick is discussed. © 1995 Chapman & Hall.

Pharmacological antagonism of Anchietia salutaris extracts on the contraction induced by prostaglandin D2 and U46619 in guinea-pig lung parenchymal strips

Anchietia salutaris tea is traditionally used in Brazil to treat allergies, suggesting it contains compounds with antagonistic activity on the allergic mediators. We have evaluated extracts and semi-purified fractions of Anchietia salutaris as a source of compounds having this type of antagonism on the contraction induced in guinea-pig lung parenchymal strips and on platelet aggregation and shape change. After 10 min pre-incubation dichloromethane extracts containing 30 or 100 μg mL-1 inhibited the contraction induced by prostaglandin D2 (PGD2) in guinea-pig lung parenchymal strips with dose ratios (DR) of 0.76 ± 0.14 and 0.93 ± 0.19, respectively; the amount of inhibition depended both on the concentration and on the time of preincubation (DR after 30 min pre-incubation was 1.21 ± 0.51). The dichloromethane extract and its semi-purified fractions also inhibited the contractions induced by U46619, a more potent, stable, synthetic agonist of thromboxane A2 (TxA2) prostanoid (TP) receptors, the receptors acted upon by PGD2 to produce lung contractions. The dichloromethane extract did not inhibit the lung parenchymal contractions induced by histamine, leukotriene D4 (LTD4) or platelet-activating factor (PAF). Platelet aggregation induced by U46619, adenosine 5'-diphosphate (ADP) or PAF was not inhibited by the dichloromethane extract. Indeed, the extract potentiated platelet aggregation induced by low concentrations of these agonists and also potentiated the shape change induced by U46619. These results imply that the dichloromethane extract of Anchietia salutaris and its semipurified fractions contain an active principle that competitively inhibits TxA2 TP receptors, the stimulation of which causes lung parenchymal contraction. The inhibition seems to be selective for this receptor subtype, because the extract fails to inhibit platelet aggregation or shape change. This provides additional support of earlier reports suggesting the occurrence of TP receptor subtypes.

Activity of Extracts and Fatty Acids of Canavalia ensiformis (Leguminosae) Against the Symbiotic Fungus of the Leaf-Cutting Ants Atta sexdens

The inhibitory effect of leaves extracts of Carnavalia ensiformis on the development of the symbiotic fungus of the leaf-cutting ants Atta sexdens (Forel) was evaluated. The hexane extract showed highest activity at concentration of 1000 μg/mL. Chromatographic separations of this extract have led to the isolation of a mixture of fatty acids which showed the same activity of the crude extract.

Enzymes present in the thoracic gland extracts from workers and males of Apis mellifera (Hymenoptera: Apidae)

This research deals with the analysis of the enzymes present in thoracic gland extracts from newly emerged, nurse workers, forager workers, newly emerged males, and mature males of A. mellifera L. (Hymenoptera, Apoidea, Apidae). The enzymes found in larger quantities in the thoracic gland occurred in all classes of workers and are digestive. Acid phosphatase and Naphtol-AS-BI-phosphohydrolase act in protein synthesis, leucine arylamidase hydrolyses proteins and a-glucosidase actuate in the nectar processing into honey. Naphtol-AS-BI-phosphohydrolase was found in larger quantities only in workers, this suggests action in protein synthesis by the thoracic gland, b-galactosidase is in larger amounts in the newly emerged bees (workers and males) this aids in the provision of other substances to be used as an energy source when glucose or sucrose are absent. Differences between enzymatic profiles from workers and males are usually related to their colony tasks, or related to their physiological necessities per individual in specific life stages.

Enzymatic activity of hypopharyngeal gland extracts from workers of Apis mellifera (Hymenoptera, Apidae, Apinae)

This research presents a comparative study of enzymatic activity of the hypopharyngeal gland extracts from workers of Apis mellifera in three physiologic stages: newly emerged, nurse and forager workers, with the objective of contributing to the comprehension of the gland function. In order to determinate the enzymes present in the extracts, the Api Zym kit (Bio Mérieux) was used to test the activity of 19 different enzymes. The enzymes found in larger amounts only in the hypopharyngeal glands from certain individuals were the following: in newly emerged workers, the N-acetyl-down double arrow sign-glucosaminidase that may be digesting the chitin of some food ingested by the bee; in forager workers, the acid phosphatase that is likely acting in authophagic processes, the a-glucosidase, in the processing of nectar into honey, and the down double arrow sign-glucosidases, in the pollen digestion.

Enzymes in the hypopharyngeal gland extracts from workers of Scaptotrigona postica (Hymenoptera, Apinae, Meliponini) related to food storing in the colony

This study reports on research of enzymes produced by the hypopharyngeal glands, which are related to food storing in the colony, from gland extracts from nurse and forager workers of S. postica. Only the presence of the saccharase was detected in the extracts from the glands of forager workers. The results were compared to the enzymatic content of similar extracts of A. mellifera taking into account the behavioral differences among the two species.

Evaluation of antimicrobial activity of extracts of medicinal plants on three tomato phytopathogens

Antimicrobial activity of 45 extracts of medicinal plants were tested on Xanthomonas campestris pv. vesicatoria, Ralstonia solanacearum and Clavibacter michiganense subsp. michiganense. Some assays were done to verify the capability of these plants extrats to show an antibiosis. Five extracts (EAFQ, SM1, SM12, SM16, SA1) shown positive activity. The extract EAFQ expressed bactericide activity on C. michiganense. It suggests the possibility of using these actives substances in natura or as a model to synthesize industrialized products, intendind field utilization.

Anti-genotoxic effect of aqueous extracts of sun mushroom (Agaricus blazei Murill lineage 99/26) in mammalian cells in vitro

The sun mushroom is the popular name for the Agaricus blazei Murill fungus, a mushroom native to south-eastern Brazil, which has been frequently used in popular medicine mainly in the form of tea to treat various ailments (stress, diabetes, etc.). In the present study, the genotoxic and/or anti-genotoxic effects ofA. blazei on mammalian cells in culture was assessed by checking the increase or reduction of micronucleus (MN) frequency and comets. The sun mushroom (lineage 99/26) was used as aqueous extracts prepared (2.5%) at three different temperatures (60, 25 and 4°C). The in vitro micronucleus (MN) test in binucleated cells and comet assay were used in V79 cells cultivated in HAM-F10+DMEM medium (1:1), supplemented with 10% of fetal bovine serum. The experiments were divided into four treatment types: 1. Negative control; 2. Positive control with MMS; 3. Treatments with the three forms of extracts (60, 25 and 4°C); and 4. Treatments with the extracts in different associations (simultaneous, pre-treatment, post-treatment and simultaneous after pre-incubation for 1 h) with MMS. None of the A. blazei extracts show genotoxic activity. In the comet assay no protecting effect was found. The results obtained in the MN test showed that the three forms of extracts used had protective activity, suggesting that the compound or active ingredients of A. blazei are always present in these extracts. The greater protective efficiency of the simultaneous treatment and simultaneous treatment with pre-incubation mixture with MMS suggests that the extracts have an antimutagenic action of the desmutagenic type. © 2002 Elsevier Science Ltd. All rights reserved.

Non-mutagenic or genotoxic effects of medicinal aqueous extracts from the Agaricus blazei mushroom in V79 cells

Agaricus blazei Murill is a mushroom largely consumed due to its medicinal properties. Effects of aqueous extract from its lineage AB97/11 in 2 fruiting body development stages (closed and opened pileus) were evaluated on chinese hamster V79 cells using cytokinesis blocking micronucleus (CBMN) and comet assays. The cells were treated at 0.15% concentration of aqueous extract prepared at different temperatures: ice-cold (4°C), room temperature (21°C) and warm (60°C). The extracts were applied in mutagenicity and antimutagenicity protocols (simultaneous, pre-incubation and continuous). The results showed that the aqueous extracts of Agaricus blazei lineage AB97/11 obtained at the 3 temperatures and both development stages did not present mutagenic or antimutagenic effect in V79 cells either in CBMN or comet assay.

Effects of crude extracts of Agaricus blazei on DNA damage and on rat liver carcinogenesis induced by diethylnitrosamine

The effects of crude extracts of the mushroom Agaricus blazei Murrill (Agaricaceae) on both DNA damage and placental form glutathione S-transferase (GST-P)-positive liver foci induced by diethylnitrosamine (DEN) were investigated. Six groups of adult male Wistar rats were used. For two weeks, animals of groups 3 to 6 were treated with three aqueous solutions of A. blazei (mean dry weight of solids being 1.2, 5.6, 11.5 and 11.5 mg/ml, respectively). After this period, groups 2 to 5 were given a single ip injection 200 mg/kg DEN and groups 1 and 6 were treated with 0.9% NaCl. All animals were subjected to 70% partial hepatectomy at week five and sacrificed 4, 24 and 48 h or 8 weeks after DEN or 0.9% NaCl treatments (10th week after the beginning of the experiment). The alkaline comet assay and GST-P-positive liver foci development were used to evaluate the influence of the mushroom extracts on liver cell DNA damage and on the initiation of liver carcinogenesis, respectively. Previous treatment with the highest concentration of A. blazei (11.5 mg/ml) significantly reduced DNA damage, indicating a protective effect against DEN-induced liver cytotoxicity/genotoxicity. However, the same dose of mushroom extract significantly increased the number of GST-P-positive liver foci.

Toothbrushing with vegetable oil: a clinical and laboratorial analysis.

The dentifrices currently available in the marketplace contain many anticariogenic substances, fluoride and abrasives aimed to better clean the dental surface, remove dental plaque, improve salivary flow and its buffer capacity and reduce colonies of bacteria such as S. mutans, the causative agent of dental caries. The objective of this study was to evaluate the possibility of adequately removing dental plaque using an experimental almond oil dentifrice (Titoil) with no abrasives or antiplaque agents. This study was carried out with 80 volunteers, all of them 18-year-old recruits from the military training school of Araçatuba -- SP. Saliva sampling and dental plaque disclosing were undertaken both before and after 28 days of toothbrushing with a low abrasive dentifrice (Group 1: 40 volunteers) or with Titoil (Group 2: 40 volunteers). Statistical analysis of the results revealed that the experimental dentifrice (Titoil) did not interfere with salivary flow and reduced dental plaque more than the low abrasive dentifrice, improved the salivary buffer capacity and decreased salivary S. mutans (Caritest-SM) as much as regular dentifrices. It was concluded that if the dental industry replaces abrasive by vegetable oil in dentifrices, these will be more effective in maintaining oral health and will cause less dental abrasion.

Toxicity of sesame extracts to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Extracts of different sesame plant (Sesamum indicum, Linnaeus) organs were tested through ingestion and contact experiments to investigate their toxicity to Atta sexdens rubropilosa (Forel) workers. Dichloromethane extracts of seeds were toxic to the leaf-cutting ants and the factor responsible for the toxicity does not show seasonal occurrence.

Bioatividade de plantas medicinais no controle de Sclerotinia isolado de Stevia rebaudiana (Bert.) Bertoni

The microorganism Sclerotinia was isolated from roots of Stevia rebaudiana (Bert.) Bertoni in plantations in the northwest of Parana and submitted to the cultivation in the presence of extracts and vegetable balsams of Tarragon (Artemisia draconculus), Thyme (Thymus vulgaris), Manjerona (Origanum majorona), Mint citrata (Mintpiperita var. citrata), Purple Basil (Ocimum basilicum L.), Andiroba (Carapa guanensis) and Copaíba (Copaifera reticulata Ducke). The first five oils were extracted by steam drags, after the drying of the vegetable in greenhouse with circulation of air at 45°C. The last two were used in natura. A suspension (100ìl) of fungus previously cultivated, was added to each plate. The results show that after 7 days of incubation the thyme oils 10ìl, purple basil 25ìl, manjerona 25ìl, mint citrata 50ìl, tarragon 50ìl were capable to inhibit the growth of Sclerotinia, while the andiroba oil only reached this result with 200ìl. The copaiba balsam, even in the concentration of 500ìl, was unable to inhibit the growth of the microorganism.

Genotoxic and antigenotoxic effects of organic extracts of mushroom Agaricus blazei Murrill on V79 cells

Agaricus blazei Murrill, popularly known as the sun mushroom, is a native mushroom in SP, Brazil, that has been widely used in the treatment of cancer and many other pathologies in different parts of the world. A water-soluble protein-polysaccharide complex (1 → 6)β-D-glucan has been isolated from its fruiting body that showed immune-modulation activity. From organic extracts, linoleic acid has been isolated and determined to be the main substance with antimutagenic activity. Using both the micronucleus (MN) and comet (single cell microgel electrophoresis) assays, this study determined the genotoxic and antigenotoxic potential of A. blazei (AB) obtained from commercial sources or the following strains: a) strains AB 97/29 (young and sporulated phases); b) a mixture taken from AB 96/07, AB 96/09 and AB 97/ 11 strains; and c) commercial mushrooms from Londrina, PR and Piedade, SP, designated as AB PR and AB SP, respectively. The extracts from these mushrooms were isolated in chloroform:methanol (3:1) and used in vitro at three different concentrations. V79 cells (Chinese hamster lung cells) were exposed to the extracts under pre-, simultaneous and post-treatment conditions, combined with methyl methanesulfonate (MMS). Under the circumstances of this study, these organic extracts did not show any genotoxic or mutagenic effects, but did protect cells against the induction of micronuclei by MMS. Copyright by the Brazilian Society of Genetics.