Propolis Standardized Extract (EPP-AF (R)), an Innovative Chemically and Biologically Reproducible Pharmaceutical Compound for Treating Wounds

The aim of this study was to develop a formulation, containing the propolis standardized extract (EPP-AF (R)), which can assist in the healing of skin lesions. To achieve this objective the antimicrobial activity and chemical composition of the propolis extract was determined. The final product was subjected to in vitro and in vivo pre-clinical evaluation. The broth macrodi-lution method was used to determine the antimicrobial activity of the extracts and formulations against the microorganisms most commonly found in burns, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. Wistar rats with puncture wounded skin were used to evaluate the wound healing properties of propolis. The results of chemical and biological characterization demonstrated the batch-to-batch reproducibility of the standardized extract which is an unprecedented result. The antimicrobial and wound healing activity of the pharmaceutical studied showed the best results when samples contain 3.6% propolis, suggesting that this is the most promising composition.

Humoral and cell-mediated immunity in large non-toxic multinodular goitre

Humoral and cell-mediated immunity was investigated in fourteen patients with non-toxic multinodular goitre and ten healthy controls by in vitro methods. These included determination of sheep erythrocyte and complement rosette-forming cells in the peripheral blood, immunoglobulin levels, titres of thyroglobulin and microsomal antibodies and migration inhibition test using thyroid extract and phytohemagglutinin. When compared with controls the patients showed high IgA levels and positive response to thyroid antigen in the leucocyte migration inhibition test. There was no correlation between the leucocyte migration results and the presence of auto-antibodies. These findings indicate a possible role of cell-mediated immunity in non-toxic multinodular goitre.

Immunological disturbances in toxic multinodular goitre and active Graves' disease

Cell-mediated and humoral immunity were studied in seventeen patients with toxic multinodular goitre, ten with active Graves' disease and fourteen healthy controls. The study included determination of sheep erythrocyte and complement rosette-forming cells in peripheral blood, immunoglobulin levels, titres of microsomal antibodies and migration inhibition test using thyroid extract and phytohaemagglutinin. When compared with controls the patients showed a positive response to thyroid antigen in the leucocyte migration inhibition test. Microsomal antibodies were detected in seven out of ten active Graves' disease patients against two out of seventeen of those with toxic multinodular goitre. Significantly increased IgG and IgA and decreased IgM levels were found only in the toxic multinodular group. These data provide further evidence for immunological disturbances in toxic multinodular goitre.

Western blot analysis of tick antigens from a Rhipicephalus sanguineus unfed larval extract and identification of antigenic sites in tick sections using immunohistochemistry. A comparative study between resistant and susceptible host species

Most parasite-host relationships are characterized by the development of resistance by the host, thus limiting the number of parasites. However, some cases are very unusual. In the relationship of the domestic dog with the brown dog-tick Rhipicephalus sanguineus this does not occur, whereas guinea pigs develop efficient resistance. Sera from domestic dogs, crab-eating foxes and guinea pigs collected before and after infestation with R. sanguineus ticks, and after immunization with a whole tick adult or larval homogenate, were used in Western blot analysis to compare and identify potential important antigens from a tick larval homogenate. The same sera were tested in an indirect immunohistochemistry assay in an attempt to compare relevant antigenic sites on histological tick sections. The immunoblotting displayed antigens recognized only by the guinea pigs, as well as several shared antigens between host species, depending on the kind of immunization. Immunohistochemistry revealed probable antigenic sites on the cells and tissues of ticks, which varied depending on the kind of immunization (infestation or vaccination) and the animal species involved.

Application of on-line C30 RP-HPLC-NMR for the analysis of flavonoids from leaf extract of Maytenus aquifolium

The application of on-line C30-reversed-phase high-pressure liquid chromatography-nuclear magnetic resonance spectroscopy is described for the analysis of tetraglycosylated flavonoids in aqueous and hydroalcoholic extracts of the leaves of Maytenus aquifolium (Celastraceae). Triacontyl stationary phases showed adequate separation for on-line 1H-NMR measurements at 600 MHz and allowed the characterisation of these flavonoids by detection of both aromatic and anomeric proton signals. Copyright (C) 2000 John Wiley and Sons, Ltd.

Elevation of boiling point of coffee extract

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

Enzymatic activity of the hypopharyngeal gland extract from workers and males of Scaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponini)

The objective of this research was to contribute to elucidation of the function of the hypopharyngeal glands of S. postica in enzyme production, using the Api Zym kit (Bio Mérieux). Dealing with a comparative analysis between the enzymatic content of the hypopharyngeal gland extracts from newly emerged, nurse and forager workers, as well as, newly emerged males and males mature for mating of S. postica. The hypopharyngeal glands from nurse workers of Apis mellifera, that produce part of the royal jelly, were used for comparison. While in A. mellifera, the hypopharyngeal glands are present only in workers, in S. postica, the hypopharyngeal glands are present and functioning in all adult individuals of the colony. The higher enzymatic activity was observed in the hypopharyngeal gland extracts from nurse workers and may be related to a larger demand for energy, compared to other individuals. The occurrence of large quantities of leucine arylamidase in all individuals may mean that protein processing is happening.

A novel biological activity for galectin-1: Inhibition of leukocyte-endothelial cell interactions in experimental inflammation

Galectin-1 (Gal-1), the prototype of a family of β -galactoside-binding proteins, has been shown to attenuate experimental acute and chronic inflammation. In view of the fact that endothelial cells (ECs), but not human polymorphonuclear leukocytes (PMNs), expressed Gal-1 we tested here the hypothesis that the protein could modulate leukocyte-EC interaction in inflammatory settings. In vitro, human recombinant (hr) Gal-1 inhibited PMN chemotaxis and trans-endothelial migration. These actions were specific as they were absent if Gal-1 was boiled or blocked by neutralizing antiserum. In vivo, hrGal-1 (optimum effect at 0.3 μg equivalent to 20 pmol) inhibited interleukin-1β-induced PMN recruitment into the mouse peritoneal cavity. Intravital microscopy analysis showed that leukocyte flux, but not their rolling velocity, was decreased by an anti-inflammatory dose of hrGal-1. Binding of biotinylated Gal-1 to resting and post-adherent human PMNs occurred at concentrations inhibitory in the chemotaxis and transmigration assays. In addition, the pattern of Gal-1 binding was differentially modulated by PMN or EC activation. In conclusion, these data suggest the existence of a previously unrecognized function of Gal-1, that is inhibition of leukocyte rolling and extravasation in experimental inflammation. It is possible that endogenous Gal-1 may be part of a novel anti-inflammatory loop in which the endothelium is the source of the protein and the migrating PMNs the target for its anti-inflammatory action.

Water-soluble extract of Coleus barbatus modulates weight gain, energy utilization and lipid metabolism in secondary biliary cirrhosis: an experimental study in young rats.

PURPOSE: To test if a water extract of Coleus barbatus (WEB) has any effect on weight gain, food energy utilization and lipid metabolism in young rats with obstructive cholestasis. METHODS: Forty 21 day old (P21) Wistar rats, in groups of 10, were submitted to one of the following treatments: a sham operation with daily water or WEB administration, double ligature and resection of the bile duct with daily water or WEB administration. At P49 they were submitted for euthanasia when the following were determined: ingested feed (IF), energy utilization (EU) and weight gain (WG) from P29 to P49, together with total serum cholesterol (TC) and triacylglycerol (TG) concentrations, liver wet weight (LWW) and fat content (LFC). Two Way ANOVA and the S.N.K. test for paired comparisons were employed to study the effects of cholestasis and those of WEB and their interactions (p < or = 0.05). RESULTS: Cholestasis, independently of WEB, and WEB, independently of cholestasis both reduced IF, EU, and WG but there was no significant interaction between the two factors. Cholestasis, independently of WEB, increased LWW, LFC, the TC and TG The WEB, independently of cholestasis, reduced these values, and there was a significant interaction between the two factors; such that these effects were more accentuated in animals with cholestasis. CONCLUSION: The WEB reduced IF, WG, and EU, both in the presence and absence of cholestasis in the same proportion. It also partially inhibited the increase in LWW, LFC, TC and TG caused by cholestasis.

Evaluation of Strychnos pseudoquina St. Hil. leaves extract on gastrointestinal activity in mice

Strychnos pseudoquina St. Hil. (Loganiaceae) was investigated for its ability to protect the gastric mucosa against injuries caused by non-steroidal anti-inflammatory drugs (piroxicam) and a necrotizing agent (HCl/EtOH) in mice. The MeOH extract and enriched alkaloidic fraction (EAF) provided significant protection in experimental models wheer used at doses of 250 and 1000 mg/kg. In vivo tests were carried out to evaluate for possible toxic effects and no mortality was observed up to the 5 g/kg dose level. Phytochemical investigation led to the isolation of a new indole alkaloid, which elucidated the observed pharmacological effects. © 2005 Pharmaceutical Society of Japan.

Evaluation of antidiarrhoeal effects of Psidium guajava L. (Myrtaceae) aqueous leaf extract in mice

A crude aqueous extract of the leaves of the guava tree, Psidium guajava L. (Myrtaceae), were studied for antidiarrhoeal effects, to place one of its traditional medical uses. The extract induced a decrease in the propulsive movements of the intestinal contents in mice. These findings suggested that an aqueous extract of guava leaves may be used as an effective treatment for non-specific diarrhoea in folk medicine.

In vivo clastogenicity assessment of the Austroplenckia populnea (Celastraceae) leaves extract using micronucleus and chromosome aberration assay

The clastogenic effect of the A. populnea leaves extract was tested in vivo on bone marrow cells of Wistar rats by evaluating the induction of chromosome aberrations and micronuclei induction on polychromatic erythrocytes. The extract was administered by gavage at doses of 300, 600 and 900mg/kg body weight. Experimental and control animals were submitted to euthanasia 24 h after the treatment. Under the conditions used, A. populnea leaves extract did not induce decrease in mitotic index and did not induce a statistically significant increase in the mean number of micronucleated polychromatic erythrocytes or chromosome aberrations in the bone marrow cells of Wistar rats. © 2007 The Japan Mendel Society.

Assessment of the potential clastogenic/aneugenic risk of Casearia sylvestris extract using in vivo assays

Casearia sylvestris (Flacourtiaceae) is a plant which grows in wild and has been widely used in folk medicine. In this study, clastogenic/aneugenic properties of Casearia sylvestris crude ethanolic extract were evaluated using in vivo chromosomal aberrations (CAs) and micronucleus (MN) assays in rodents. The animals were treated by gavage with 3 concentrations of the extract: 150, 300 and 500 mg/kg body weight. Bone marrow cells from Wistar rats were collected 24 h after having been submitted to the MN and CAs test. Peripheral blood cells from Swiss mice were collected 48 and 72 h after having been submitted to the MN test. The results show that C. sylvestris extract does not induce a significant increase in mean values for micronucleated polychromatic erythrocytes (MNPCE) in Swiss mice and Wistar rats, or CAs in rat bone marrow cells, at the 3 tested doses, indicating that the extract showed no clastogenic/aneugenic effects on chromosomes of the rodent cells tested. © 2007 The Japan Mendel Society.

Avoiding leukocyte contamination and early platelet activation in platelet-rich plasma.

The objective of this study was to describe a new platelet-rich plasma (PRP) protocol with a reduced concentration of leukocytes and intact platelets. We collected 8 mL of venous blood (VB) from marginal ear veins of 10 male New Zealand white rabbits in acid dextrose citrate Vacutainer tubes. Tubes were centrifuged at 302g for 10 minutes. All plasma was collected in plastic tubes to avoid buffy-coat contamination and centrifuged at 2862g for 5 minutes. A 10% calcium chloride activator (10 PRP:2 CaCl2) was added to the lower third of this plasma (PRP), and the PRP gel was obtained. Mean platelet count was 317.7 x 10(3) +/- 39.9/microL in VB and 1344.9 x 10(3) +/- 347.5/microL in PRP. Leukocyte counts were 3.96 x 10(3) +/- 2.01/microL and 0.46 x 10(3) +/- 0.45/microL in VB and PRP, respectively. Mean platelet enrichment was 327.4 +/- 97.8%. All differences were statistically significant (P > .05). This protocol is practical and reproducible, resulting in a high concentration of intact platelets to help tissue repair and low levels of leukocytes.

Antioxidant action of Rosemary extract in soybean oil submitted to thermoxidation

This work was aimed at evaluating the antioxidant activity of rosemary extract added to soybean oil in thermoxidation conditions. Purified soybean oil, refined soybean oil and refined soybean oil containing 1,000 mg/kg rosemary extract were heated at 180°C. The oxidation of the samples was evaluated after 0, 2.5, 5, 7.5 and 10 hours of thermoxidation by means of oxidative stability determination, total polar compounds and conjugated dienes. The purified oil differed significantly from the refined oil, mainly in relation to oxidative stability due the removal of the natural antioxidants. Rosemary extract presented antioxidant effects at high temperatures. After 10 hours of heating, 1,000 mg/kg rosemary extract added to the refined soybean oil significantly increased the oil oxidative stability from 7.52 to 13.5 hours and decreased the formation of polymers and decomposing products measured through the polar rates from 17.35 to 7.99%. The build up of primary oxidation products gauged through diene rates also decreased from 1.61 to 0.80%. Rosemary extract could be recommended as an alternative antioxidant.