Pedro Luiz Rosalen

University of Campinas, Conceição de Campinas, São Paulo, Brazil

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Publications (113)167.86 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Objective: to elucidate the mechanisms involved in the anti-inflammatory properties of Brazilian red propolis Method: different concentrations of ethanolic extract of Brazilian red propolis (EEBRP) were added to RAW 264.7 macrophages after activation with LPS, and NO production and cell viability determined. Production of TNF-α, IL1β, TGF-β, IL-4, IL-6, IL-10, IL-12, GM-CSF, IFN-Ɣ and expression of genes related to cytokines production and nitric oxide , PI3K-AKT and signal transduction pathway were evaluated by ELISA and RT-qPCR, respectively. Differences were determined by one-way ANOVA followed by Tukey-Kramer Result: EEBRP inhibited NO production by 69% without affecting cell viability at the concentration of 50 µg/ml when compared with control cells (treated with vehicle) (p<0.05). Levels of IL-12, GM-CSF, IFN-Ɣ, IL-1β, IL-4, IL-10 and TGF-β in cell supernatant were reduced and of TNF- α and IL-6 increased with the addition of EEBRP (p<0.05). EEBRP induced alteration in expression of 57 genes. Genes involved in Toll-like response; oxide nitric sintetase production (all types), Nf-KB and MAPK signaling pathways such as Tirap, Pdk1, Pak1, Nfkb1, Mtcp1, Gsk3b, Fos, Elk1, Il1β, Il1f9 were negatively regulated by EEBRP(fold-change rate > 5; p<0.05) Conclusion: EEBRP presents antiinflammatory properties by altering signaling pathways leading to reduction in production of pro-inflammatory factors.
    2014 IADR/PER Congress; 09/2014
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    ABSTRACT: Objective: recent data have shown that Brazilian red propolis presents immunemodulatory properties, which led us to elucidate the mechanisms involved in the anti-inflammatory properties of one of its components, Neovestitol Method: neovestitol was obtained from ethanolic extract of Brazilian red propolis after bio-guided fractionation coupled with GC-MS analysis. Different concentrations of Neovestitol were added to RAW 264.7 macrophages after activation with LPS. After 48 hours, NO production and cell viability were determined. Production of TNF-α, IL1β, TGF-β, IL-4, IL-6, IL-10, IL-12, GM-CSF, IFN-Ɣ and expression of genes related to cytokines production, nitric oxide , PI3K-AKT and signal transduction pathways were evaluated by ELISA and RT-qPCR, respectively. Differences were determined by one-way ANOVA followed by Tukey-Kramer Result: Neovestitol inhibited NO production by 63% without affecting cell viability at the concentration of 60 µg/ml when compared with control cells (treated with vehicle) (p<0.05). Levels of GM-CSF, IFN- Ɣ, IL-1 β, IL-4, TNF- α and IL-6 in cell supernatant were reduced and of IL-10 increased with the addition of neovestitol (p<0.05). Neovestitol induced alteration in the expression of 43 genes. Genes involved in Toll-like response; nitric oxide sintetase production (all types), Nf-KB and Il-1 signaling pathways such as Bad, Chuk, Elk1, Ifnb1, Il1b, Lif, Calm1, Capns1, Egr1, Nox1, Scd2 and Scd3 were negatively regulated by neovestitol (fold-change rate > 5; p<0.05). Furthermore, its role in inhibition of leukocyte transmigration was suggested by inhibition of expression of Nox1 and in the regulation of fatty acid associated genes such as Scd2 e Scd3 Conclusion: Neovestitol may represent a new antiinflammatory agent by inhibiting signaling pathways and consequently reducing the production of pro-inflammatory factors and by interfering in leukocyte transmigration
    2014 IADR/PER Congress; 09/2014
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    ABSTRACT: The aim of this study was to evaluate the anti-inflammatory activity of Petit Verdot Extract and hexane, chloroform and ethyl acetate fractions obtained from grape pomace, in addition to identifying active compounds. The PVE and EAF reduced significantly paw edema and neutrophil migration when compared with control groups. The PVE reduced levels of TNF-α and IL1-β in the peritoneal fluid, whereas the EAF did not reduce cytokines significantly. Two hydroxybenzoic acids, two proanthocyanidins, three flavan-3-ol monomers and three anthocyanins were identified in the PVE and EAF by LC-MS/MS. The stilbene trans-resveratrol was identified only in the EAF. The phenolic compounds were quantified using HPLC-DAD analysis, except for the stilbenes, which were not sensible for the detection by the method. Since PVE and EAF showed significantly anti-inflammatory effects and high concentration of phenolic compounds, we concluded that Petit Verdot pomace could be an interesting source of anti-inflammatory bioactives.
    Journal of Functional Foods 05/2014; 8:292–300. · 2.63 Impact Factor
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    ABSTRACT: Purpose: To compare the pharmacokinetic profiles and to evaluate the bioequivalence of two commercial amoxicillin suspension formulations (500 mg/5 mL AMOXIL®, reference formulation and AMOXI-PED®, test formulation) in healthy Brazilian volunteers. Methods: Under fasting condition, 25 volunteers (13 males and 12 females) were included in this randomized, open-label, two-period crossover (1-week washout interval) bioequivalence study. Blood samples were collected at pre-dose (0 hour) and 0.5, 1, 1.33, 1.66, 2, 2.5, 3, 4, 6, 8, and 12 hours after drug ingestion. Pharmacokinetic parameters (Cmax, tmax, t1/2, AUC0-tlast, and AUC0-∞) were calculated from plasma concentrations for both formulations in each subject. Results: Arithmetic mean values of the pharmacokinetic parameters were: Cmax = 12.004 (± 2.824) μg×mL-1; tmax = 1.118 (± 0.396) h; t1/2 = 1.226 (± 0.179) h; AUC0-tlast = 29.297 (± 6.007) μg×h×mL-1; and AUC0-∞ = 29.299 (± 6.007) μg×h×mL-1 for reference formulation and Cmax = 11.456 (± 2.825) μg×mL-1; tmax = 1.331 (± 0.509) h; t1/2 = 1.141 (± 0.133) h; AUC0-tlast = 28.672 (± 5.778) μg×h×mL-1; and AUC0-∞ = 28.693 (± 5.796) μg×h×mL-1 for test formulation. The confidence intervals (90% CI) for reference and test formulations were, respectively, 90.74 - 100.46% for Cmax and 93.62 - 103.61% for AUC0-t. Conclusion: Based on the results, both formulations of amoxicillin evaluated in this study were considered bioequivalent according to FDA and ANVISA/Brazil criteria.
    International journal of clinical pharmacology and therapeutics 04/2014; · 1.20 Impact Factor
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    ABSTRACT: Oral candidiasis is an opportunistic fungal infection of the oral cavity with increasingly worldwide prevalence and incidence rates. Novel specifically-targeted strategies to manage this ailment have been proposed using essential oils (EO) known to have antifungal properties. In this study, we aim to investigate the antifungal activity and mode of action of the EO from Coriandrum sativum L. (coriander) leaves on Candida spp. In addition, we detected the molecular targets affected in whole-genome expression in human cells. The EO phytochemical profile indicates monoterpenes and sesquiterpenes as major components, which are likely to negatively impact the viability of yeast cells. There seems to be a synergistic activity of the EO chemical compounds as their isolation into fractions led to a decreased antimicrobial effect. C. sativum EO may bind to membrane ergosterol, increasing ionic permeability and causing membrane damage leading to cell death, but it does not act on cell wall biosynthesis-related pathways. This mode of action is illustrated by photomicrographs showing disruption in biofilm integrity caused by the EO at varied concentrations. The EO also inhibited Candida biofilm adherence to a polystyrene substrate at low concentrations, and decreased the proteolytic activity of Candida albicans at minimum inhibitory concentration. Finally, the EO and its selected active fraction had low cytotoxicity on human cells, with putative mechanisms affecting gene expression in pathways involving chemokines and MAP-kinase (proliferation/apoptosis), as well as adhesion proteins. These findings highlight the potential antifungal activity of the EO from C. sativum leaves and suggest avenues for future translational toxicological research.
    PLoS ONE 01/2014; 9(6):e99086. · 3.53 Impact Factor
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    ABSTRACT: The aim of the present study was to perform an in vitro analysis of the antimicrobial and antiproliferative potential of an extract from Anadenanthera colubrina (Vell.) Brenan (angico) and chemically characterize the crude extract. Antimicrobial action was evaluated based on the minimum inhibitory concentration (MIC), minimum bactericidal/fungicidal concentration, and the inhibition of formation to oral biofilm. Cell morphology was determined through scanning electron microscopy (SEM). Six strains of tumor cells were used for the determination of antiproliferative potential. The extract demonstrated strong antifungal activity against Candida albicans ATCC 18804 (MIC = 0.031 mg/mL), with similar activity found regarding the ethyl acetate fraction. The extract and active fraction also demonstrated the capacity to inhibit the formation of Candida albicans to oral biofilm after 48 hours, with median values equal to or greater than the control group, but the difference did not achieve statistical significance (P > 0.05). SEM revealed alterations in the cell morphology of the yeast. Regarding antiproliferative activity, the extract demonstrated cytostatic potential in all strains tested. The present findings suggest strong antifungal potential for Anadenanthera colubrina (Vell.) Brenan as well as a tendency toward diminishing the growth of human tumor cells.
    Evidence-based complementary and alternative medicine : eCAM. 01/2014; 2014:802696.
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    ABSTRACT: The present study examined the influences of the neovestitol-vestitol (NV) containing fraction isolated from Brazilian red propolis on the development of biofilm and expression of virulence factors by Streptococcus mutans using saliva-coated surfaces of hydroxyapatite. In addition, NV was tested in a rodent model of dental caries to assess its potential effectiveness in vivo. Topical applications of NV (800 μg ml(-1)) significantly impaired the accumulation of biofilms of S. mutans by largely disrupting the synthesis of glucosyltransferase-derived exopolysaccharides and the expression of genes associated with the adaptive stress response, such as copYAZ and sloA. Of even greater impact, NV was as effective as fluoride (positive control) in reducing the development of carious lesions in vivo. NV is a promising natural anti-biofilm agent that targets essential virulence traits in S. mutans, which are associated with the formation of cariogenic biofilm and the subsequent onset of dental caries disease.
    Biofouling 10/2013; · 3.40 Impact Factor
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    ABSTRACT: Guava pomace is an example of the processing waste generated after the manufacturing process from the juice industry that could be a source of bioactives. Thus, the present investigation was carried out in order to evaluate the anti-inflammatory and antinociceptive potential and determinate the main phenolic compounds of a guava pomace extract (GPE). The anti-inflammatory activity was evaluated by carrageenan, dextran, serotonin, histamine-induced paw edema and neutrophils migration in the peritoneal cavity models. Acetic acid-induced abdominal writhing and formalin test were performed to investigate the antinociceptive effects. In addition, the content of total phenolic and of individual phenolic compounds was determined by GC/MS. GPE showed anti-inflammatory activity by carrageenan, dextran, serotonin, histamine-induced paw edema and neutrophils migration in the peritoneal cavity models (p < 0.05). GPE also demonstrated antinociceptive activity by acetic acid-induced abdominal writhing and formalin test (p < 0.05). The total phenolic value was 3.40 +/- 0.09 mg GAE/g and epicatechin, quercetin, myricetin, isovanilic and gallic acids were identified by GC/MS analysis. The presence of bioactive phenolic compounds as well as important effects demonstrated in animal models suggest that guava pomace could be an interesting source of anti-inflammatory and analgesic substances.
    BMC Complementary and Alternative Medicine 09/2013; 13(1):235. · 2.08 Impact Factor
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    ABSTRACT: The objective of this study was to evaluate anti-inflammatory and antimicrobial activities of neovestitol and vestitol isolated from Brazilian red propolis (BRP). BRP ethanolic extract (EEP), neovestitol and vestitol were evaluated by anti-inflammatory properties using a neutrophil migration assays. The antimicrobial activity was evaluated by minimal inhibitory and bactericidal concentrations (MIC and MBC) against Sreptococcus mutans, Sreptococcus sobrinus, Sthaphylococcus aureus, and Actinomyces naeslundii. Neovestitol, vestitol and EEP inhibited neutrophil migration at dose of 10 mg/Kg. Regarding antimicrobial activity, neovestitol showed MIC ranging from <6.25 to 25-50 µg/mL and MBC ranging from 25-50 to 50-100 µg/mL, while vestitol showed MIC ranging from 25-50 to 50-100 µg/mL and MBC ranging from 25-50 to 50-100 µg/mL. Both isoflavonoids, neovestitol and vestitol, are consistent bioactive compounds displaying anti-inflammatory and antimicrobial activities that can strongly act in low dose and concentration and have a promising potential to be applied on pharmaceutical and food industries.
    Journal of Agricultural and Food Chemistry 04/2013; · 3.11 Impact Factor
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    ABSTRACT: Objective: The purpose of this study was to evaluate in vitro and in vivo anti-inflammatory properties (in vitro and in vivo) of Brazilian red propolis. Method: Ethanolic extract of Brazilian red propolis (BRP) was investigated on modulation of NO production by macrophages RAW 264.7 (in vitro) and on cells viability. In vivo neutrophil migration inhibition activity induced by carrageenan was also evaluated, as well its mechanism of action such as NO pathway (by previous administration of aminoguanidine, an inhibitor of nitric oxide synthesis), TNF-α, IL-1β and IL-10 (by ELISA). The statistical analysis was performed according to one-way ANOVA followed by Tukey-Kramer for in vitro assays and ANOVA followed by Bonferroni test for in vivo assays. Result: BRP modulated NO production by macrophages at concentrations as low as 60µg/mL without affecting cells viability. BRP was also effective in preventing neutrophil migration to peritoneal cavity at a dose of 10 mg/kg (p<0.05). The in vivo inhibitory effect caused by BRP appears to be associated with inhibition of nitric oxide pathway and with decrease in production of TNF-α without affecting IL-1β (p<0.05) and IL-10 levels. Conclusion: The Brazilian red propolis may be a promising anti-inflammatory natural product, affecting nitric oxide production and cytokines profile.
    IADR/AADR/CADR General Session and Exhibition 2013; 03/2013
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    ABSTRACT: Objective: To evaluate the in vitro antimicrobial activity of essential oils (EO) and fractions against Streptococcus mutans, as well as the phytochemical profile of those presenting the best activity. Method: Twenty EO were obtained from plants (Germplasm bank/UNICAMP) by hidrodestilation. This was a bioguided study using antimicrobial assays performed by Minimum Inhibitory (MIC) and Bactericidal (MBC) Concentrations of EO and inhibition of Streptococcus mutans UA159 biofilm. Positive (0.12% Chlorhexidine) and negative (24% propylene glycol) controls were also used. Thus, the EO were selected according to the following criteria: best antimicrobial activity (MIC<250 μg /mL), extract yield, commercial availability, part of the plant from which OE were obtained and easy cultivation. Phytochemical analyses were carried out on selected EO by Thin Layer Chromatography (TLC) and Gas Chromatography/Mass Spectrometry (GC-MS) and fractionation by dry column, which resulted in fractions with different polarity. Finally, 22 fractions were generated, which also were submitted to antimicrobial assays Result: MIC values of 20 EO ranged from 31.2 to 500 μg/mL, and MBC from 62.5-1000 μg/mL. Biofilm formation inhibition was higher than 80% at concentrations above MIC for all EO tested. Considering the aforementioned criteria, the selected EO came from Aloysia gratissima, Baccharis dracunculifolia, Coriandrum sativum and Lippia sodoides. GC-MS of these four EO showed terpenes, sesquiterpenes, aldehydes and alcohols. MIC values of active fractions from selected EO ranged from 31.2 to 125 μg/mL and MBC from 31.2 to 250 μg/mL. All active fractions inhibited more than 80% of mutans biofilm formation. Conclusion: All EO studied showed strong antimicrobial and antibiofilm activities against mutans. Four of them were selected as the best ones, whose active fractions presented chemical compounds related to antimicrobial activity. These EO could be a promising source to discovery of new bioactive molecules against S. mutans
    IADR/AADR/CADR General Session and Exhibition 2013; 03/2013
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    ABSTRACT: BACKGROUND: Geopropolis is a type of propolis containing resin, wax, and soil, collected by threatened stingless bee species native to tropical countries and used in folk medicine. However, studies concerning the biological activity and chemical composition of geopropolis are scarce. In this study, we evaluated the antimicrobial and antiproliferative activity of the ethanolic extract of geopropolis (EEGP) collected by Melipona scutellaris and its bioactive fraction against important clinical microorganisms as well as their in vitro cytotoxicity and chemical profile. METHODS: The antimicrobial activity of EEGP and fractions was examined by determining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against six bacteria strains as well as their ability to inhibit Streptococcus mutans biofilm adherence. Total growth inhibition (TGI) was chosen to assay the antiproliferative activity of EEGP and its bioactive fraction against normal and cancer cell lines. The chemical composition of M. scutellaris geopropolis was identified by reversed-phase high-performance liquid chromatography and gas chromatography--mass spectrometry. RESULTS: EEGP significantly inhibited the growth of Staphylococcus aureus strains and S. mutans at low concentrations, and its hexane fraction (HF) presented the highest antibacterial activity. Also, both EEGP and HF inhibited S. mutans biofilm adherence (p < 0.05) and showed selectivity against human cancer cell lines, although only HF demonstrated selectivity at low concentrations. The chemical analyses performed suggest the absence of flavonoids and the presence of benzophenones as geopropolis major compounds. CONCLUSIONS: The empirical use of this unique type of geopropolis by folk medicine practitioners was confirmed in the present study, since it showed antimicrobial and antiproliferative potential against the cancer cell lines studied. It is possible that the major compounds found in this type of geopropolis are responsible for its properties.
    BMC Complementary and Alternative Medicine 01/2013; 13(1):23. · 2.08 Impact Factor
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    ABSTRACT: The aim of this study was to evaluate the activity of the ethanolic extract of geopropolis (EEGP) from Melipona scutellaris and its fractions on the modulation of neutrophil migration in the inflammatory process, and the participation of nitric oxide (NO) pathway, as well as to check the chemical profile of the bioactive fraction. EEGP and its aqueous fraction decreased neutrophil migration in the peritoneal cavity and also the interaction of leukocytes (rolling and adhesion) with endothelial cells. The levels of chemokines CXCL1/KC and CXCL2/MIP-2 were not altered after treatment with EEGP and the aqueous fraction. It was found that the injection of NO pathway antagonists abolished the EEGP and the aqueous fraction inhibitory activity on the neutrophil migration. The expression of intercellular adhesion molecule type 1 (ICAM-1) was reduced, and nitrite levels increased after treatment with EEGP and aqueous fraction. In the carrageenan-induced paw edema model, EEGP and the aqueous fraction showed antiedema activity. No pattern of flavonoid and phenolic acid commonly found in propolis samples of Apis mellifera could be detected in the aqueous fraction samples. These data indicate that the aqueous fraction found has promising bioactive substances with anti-inflammatory activity.
    Evidence-based Complementary and Alternative Medicine 01/2013; 2013:907041. · 1.72 Impact Factor
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    ABSTRACT: The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis on Streptococcus mutans biofilm. The ethanolic extract of Melipona scutellaris geopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF) possessing antimicrobial activity. The effects of HF on S. mutans UA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM). HF at 250 μ g/mL and 400 μ g/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P < 0.05) subsequently observed at SEM images, and this reduction was noticed in the amounts of extracellular alkali-soluble glucans, intracellular iodophilic polysaccharides, and proteins. In addition, the S. mutans viability (killing assay) and acid production by glycolytic pH drop were not affected (P > 0.05). In conclusion, the bioactive HF of geopropolis was promising to control the S. mutans biofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.
    Evidence-based Complementary and Alternative Medicine 01/2013; 2013:256287. · 1.72 Impact Factor
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    ABSTRACT: Culturing methods are the primary approach for microbiological analysis of plaque biofilms in rodent models of dental caries. In this study, we developed strategies for the isolation of DNA and RNA from plaque biofilms formed in vivo to analyse the viable bacterial population and gene expression. Plaque biofilm samples from rats were treated with propidium monoazide to isolate DNA from viable cells, and the purified DNA was used to quantify total bacteria and the Streptococcus mutans population via quantitative polymerase chain reaction (qPCR) and specific primers; the same samples were also analysed by counting colony-forming units (CFU). In parallel, RNA was isolated from plaque-biofilm samples (from the same animals) and used for transcriptional analyses via reverse transcription-qPCR. The viable populations of both S. mutans and total bacteria assessed by qPCR were positively correlated with the CFU data (P < 0.001; r > 0.8). However, the qPCR data showed higher bacterial cell counts, particularly for total bacteria (vs. CFU). Moreover, S. mutans proportion in the plaque biofilm determined by qPCR analysis showed strong correlation with incidence of smooth-surface caries (P = 0.0022, r = 0.71). The purified RNAs presented high RNA integrity numbers (>7), which allowed measurement of the expression of genes that are critical for S. mutans virulence (e.g. gtfB and gtfC). Our data show that the viable microbial population and the gene expression can be analysed simultaneously, providing a global assessment of the infectious aspect of dental caries. Our approach could enhance the value of the current rodent model in further understanding the pathophysiology of this disease and facilitating the exploration of novel anti-caries therapies.
    Molecular oral microbiology. 10/2012; 27(5):350-61.
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    ABSTRACT: Biological assays that have been performed on different types of Brazilian propolis have shown that type 6 propolis (G6) has a strong antimicrobial activity and a low flavonoid content. This study aimed to evaluate the correlation between the phenolic composition and the biological activities displayed by propolis G6 from the state of Bahia and green propolis, also known as type 12 (G12). The values of the flavonoids and the total phenolics in propolis G6 were different than those in propolis G12. Although the G12 variety presented greater antioxidant activity, propolis G6 proved to have greater antimicrobial and cytotoxic activities. The results showed that the phenolic compounds may not be the only compounds responsible for the biological activity. More detailed studies of the chemical composition and an assessment of the biological activity are required to establish the quality of propolis.
    Brazilian Journal of Pharmaceutical Science 09/2012; 48(3):557-564. · 0.37 Impact Factor
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    ABSTRACT: The pharmacological activity of geopropolis collected by stingless bees (important and threatened pollinators), a product widely used in folk medicine by several communities in Brazil, especially in the Northeast Region, needs to be studied. The aim of this study was to evaluate the antinociceptive activity of Melipona scutellaris geopropolis (stingless bee) using different models of nociception. The antinociceptive activity of the ethanolic extract of geopropolis (EEGP) and fractions was evaluated using writhing induced by acetic acid, formalin test, carrageenan-induced hypernociception, and quantification of IL-1β and TNF-α. The chemical composition was assessed by quantification of total flavonoids and phenolic compounds. EEGP and its hexane and aqueous fractions showed antinociceptive activity. Both EEGP and its aqueous fraction presented activity in the mechanical inflammatory hypernociception induced by the carrageenan model, an effect mediated by the inhibition of IL-1β and TNF-α. The chemical composition of EEGP and its hexane and aqueous fractions showed a significant presence of phenolic compounds and absence of flavonoids. Our data indicate that geopropolis is a natural source of bioactive substances with promising antinociceptive activity.
    Journal of ethnopharmacology 08/2012; 143(2):709-15. · 2.32 Impact Factor
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    ABSTRACT: The rodent model of dental caries has provided valuable information about the infectious character of the disease and the developmental process of carious lesions. Efforts to improve and establish caries scoring methods were performed over the years, but little has been done to refine plaque-biofilm analyses. Culturing methods are the primary approach for microbiological analysis of plaque-biofilms in rodent models of dental caries. Objectives: In this study, we developed strategies for isolation of DNA and RNA from in vivo formed plaque-biofilms to analyze the viable bacterial population and gene expression. Methods: Plaque-biofilm samples from rats were treated with propidium monoazide to isolate DNA from viable cells, and the purified DNA was used to quantify total bacteria and S. mutans population via qPCR and specific primers; the same samples were also analyzed by colony forming unit (CFU) counting. In parallel, RNA was isolated from plaque-biofilm samples (from same animals) and used for transcriptional analyses via RT-qPCR. Results: The viable population of both S. mutans and total bacteria assessed by qPCR were positively correlated with the CFU data (P<0.001; r>0.8). However, the qPCR data showed higher bacterial cell counts, particularly for total bacteria (vs. CFU). Moreover, S. mutans proportion in the plaque-biofilm determined by qPCR analysis showed strong correlation with incidence of smooth-surface caries (P=0.0022, r=0.71). The purified RNAs presented high RNA integrity numbers (>7), which allowed measurement of the expression of low abundance transcripts that are critical for S. mutans virulence (e.g. gtfB and gtfC). Conclusion: Our data show that the viable microbial population and the gene expression can be analyzed simultaneously, providing a global assessment of the infectious aspect of dental caries. Our approach could enhance the value of the current rodent model in further understanding the pathophysiology of this disease and facilitating the exploration of novel anti-caries therapies.
    06/2012
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    ABSTRACT: Objective: The purpose of this study was to evaluate the effect of Brazilian red propolis (crude extract) and the isolated active compounds on the modulation of neutrophils migration during the inflammatory process, as well as their mechanisms of action. Method: To reach this objective, ethanolic extract of Brazilian red propolis (EEP) was submitted to a bio-guided fractionation coupled with GC-MS analysis, which resulted in 2 purified compounds, neovestitol and vestitol. EEP, neovestitol and vestitol were investigated on neutrophil migration inhibition activity induced by carrageenan. Also, levels of TNF-α, IL-1β and IL-10 were measured from the peritoneal fluid collected from animal treated with the samples. The statistical analysis was performed according to one-way ANOVA followed by Bonferroni test. Result: The agents tested were effective in prevent neutrophil migration to peritoneal cavity at a dose of 10 mg/kg (p<0.05), but vestitol was particularly effective at concentrations as low as 1 mg/kg (p<0.05). The inhibitory effect caused by EEP appears to be associated with the decrease of TNF-α level and increase of anti-inflammatory IL-10 level, without affecting IL-1β (p<0.05), as verified by ELISA. Neovestitol inhibited TNF-α and IL-1β levels, but it was without effect on IL-10 (p<0.05). In contrast, vestitol increased the anti-inflammatory IL-10 level, without displaying any significant effects on TNF-α and IL-1β (p<0.05). Conclusion: The Brazilian red propolis may be a promissing anti-inflammatory natural product while neovestitol and vestitol appears to be important bioactive compounds. Future studies are needed to further characterize the mechanisms of action of these two compounds
    IADR General Session 2012; 06/2012
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    ABSTRACT: Objective: To evaluate the activity of essential oils (EO) against biofilm formed by Streptococcus mutans UA159 and determine their cytotoxicity on normal and tumor cells lines. Also, this study characterizes the chemical composition of the EO active fractions. Method: EO (Aloysia gratissima, Baccharis dracunculifolia, Coriandrum sativum, and Lippia sodoides) were obtained by hydrodistillation from plants of the germplasm collection (CPMA/CPQBA/UNICAMP). The EO were fractioned by dry column and the active fractions (AF) were selected according to biological criteria. This bioguided study included: 1) Antimicrobial assays: minimum inhibitory (MIC) and bactericidal (MBC) concentrations; inhibition and morphology analysis of S. mutans biofilm formation by scanning electron microscopy (SEM); 2) Cytotoxicity assays: in vitro study of antiproliferative activity on normal (Keratinocytes) and on seven tumor human cells lines; and 3) Chemical assays: thin layer chromatography (TLC) and gas chromatography/mass spectrometry (GC-MS) for phytochemical analysis. Positive (chlorhexidine 0.12%) and negative (propylene glycol 6.25% and 25%) controls were performed. Statistic analysis was done by non-parametric Kruskall-Wallis test. Results: MIC and MBC values obtained from all four EO and their respective AF showed strong antimicrobial activity (between 500 to 15.6 μg.mL-1). The AF of Baccharis dracunculifolia, Coriandrum sativum at 31.2 μg.mL-1 showed the best inhibition (90%) of S. mutans biofilm (p < 0.05) and SEM shows the alterations in the biofilm structures. The AF did not interfere on normal cell line proliferation, however showed specificity against some tumor cell lines evaluated in this study. Chemical analysis of AF showed mainly the presence of terpenes. Conclusion: All EO studied showed strong antimicrobial activity and two AF were selected as the best antibiofilm effect and no cytotoxicity on normal cell. Thus, these AF could be promising a discovery of new bioactive molecules against S. mutans.
    IADR General Session 2012; 06/2012

Publication Stats

1k Citations
167.86 Total Impact Points

Institutions

  • 1998–2014
    • University of Campinas
      • Faculty of Dentistry from Piracicaba
      Conceição de Campinas, São Paulo, Brazil
  • 2011
    • Chonbuk National University
      Tsiuentcheou, North Jeolla, South Korea
  • 1996–2011
    • University of Rochester
      • Center for Oral Biology
      Rochester, NY, United States
  • 2003–2009
    • University Center Rochester
      • Center for Oral Biology
      Rochester, Minnesota, United States
    • São Paulo State University
      • Departamento de Ciências Biológicas (Bauru)
      São Paulo, Estado de Sao Paulo, Brazil
  • 2008
    • Faculdade de Ciências da Saúde de São Paulo
      San Paulo, São Paulo, Brazil
  • 2007
    • State University of Ponta Grossa
      • Department of Pharmaceutical Sciences
      Ponta Grossa, Paraná, Brazil