Toxicity of Propylene Glycol Extract of Propolis on Central Nervous System and Liver in Pregnant and Neonatal Rats.

BACKGROUND: Propolis has become one of the most preferred supplements due to its beneficial biological properties. Organic (water and vegetable oils) and chemical (ethyl alcohol, propylene glycol, and glycerol) solvents are used for propolis extraction. However, the effects of these chemicals on health should be taken into account. OBJECTIVES: In this study, the effects of propolis extracts on health were evaluated. METHODS: 32 pregnant Wistar albino rats and 64 neonatal/young adults were given three different extractions of propolis (propylene glycol, water, and olive oil). Histopathological analyses were performed on the liver and brain, and blood samples were taken from the hearts of rats. RESULTS: Histopathological scoring showed that the intensity of pycnotic hepatocyte, sinusoidal dilatation, and bleeding was high in liver samples of pregnant and baby rats given propylene glycol extract of propolis (p<0.05). Propylene glycol extract caused dilatation of blood vessels and apoptosis of neurons in brain tissue. The histopathological score was significantly lower in liver and brain tissues of rats treated with water and olive oil extract compared to propylene propolis groups (p<0.05). Liver enzyme levels in the blood increased in propylene propolis rats (p<0.05). CONCLUSION: Histopathological changes and biochemical alterations may indicate that propylene glycol extracts of propolis are more toxic than olive oil and water extracts. Therefore, olive oil and water extracts of propolis are more reliable than propylene glycol extract in pregnant and infant rats.

Genotoxicity and toxicological evaluations of Brazilian red propolis oral ingestion in a preclinical rodent model.

ETHNOPHARMACOLOGICAL RELEVANCE: Brazilian red propolis is a natural product known due to its medicinal properties. The efficacy of this natural resin has been proved; however, few studies report the safety of its oral use. Some toxic effects of natural products may not be expressed in traditional use, and preclinical studies are necessary to guarantee their safety. Health regulatory agency currently requires these non-clinical studies to develop drugs and herbal medicines, including genotoxic and oral toxicity tests. AIM OF THE STUDY: Accomplish the preclinical toxicity studies of Brazilian red propolis extract (BRP) in rodents, including genotoxicity, acute and sub-chronic toxicities. MATERIAL AND METHODS: Genotoxicity assays followed the erythrocyte micronucleus test protocol in a range of 500-2000 mg/kg BRP oral treatment on male Swiss mice. After an up-and-down procedure, acute oral toxicity (single dose) was performed on female Wistar Hannover rats, reaching a 2000 mg/kg BRP oral gavage concentration. Animals were monitored periodically until 14 days and euthanized for a macroscopic necropsy analysis. The sub-chronic oral toxicity test (90 days) was achieved with 1000 mg/kg of BRP on Wistar Hannover rats (males/females). Animals were monitored to evaluated behavioral and biometrical changes, then were euthanized to perfomed hematological, biochemical, and histopathological analyses. RESULTS: No genotoxic effect of the BRP was detected. The acute toxicity indicated no toxicity of a single oral dose of 2000 mg/kg of BRP. The long-term oral toxicity performed with 1000 mg/kg of BRP altered water and food intake and the biometrics, hematological and biochemical parameters. Biochemical alterations in hepatic and renal parameters were detected only in the males. Despite the detection of biochemical alterations, no histopathological changes were detected in the organs of any group. CONCLUSIONS: BRP, at a higher dose, showed no signs of immediate toxicity. However, the obtained results suggest that the chemical composition and the intake of higher doses deserve special attention regarding possible toxicity.

Exploring the anticancer effects of standardized extracts of poplar-type propolis: In vitro cytotoxicity toward cancer and normal cell lines.

Propolis was shown to exert antimicrobial, antioxidant, anti-inflammatory, and anticancer activities. Its composition is influenced by seasonal, climatic and phytogeographic conditions. Further variability derives from the extraction methods. Multi Dynamic Extraction Method (MED) has been recently proposed to improve extracts reproducibility. Here, the cytotoxic/anticancer activity of three MED extracts of poplar-type propolis was assayed on human promyelocytic leukaemia HL60, human monocytic leukaemia THP-1, human osteosarcoma MG63, murine fibroblast L929 and human mesenchymal cells (hMSCs). As far as we are aware of, MG63 cells have never been challenged with propolis before, while few studies have so far addressed the effects of propolis on non-tumor cell lines. Consistent results were observed for all propolis preparations. The extracts turned out mildly cytotoxic toward cancer cells, in particular osteosarcoma cells (IC50: 81.9-86.7 µg/ml). Nonetheless, cytotoxicity was observed also in non-tumor L929 cells, with an even lower IC50. hMSCs demonstrated the lowest sensitivity to propolis (IC50: 258.3-287.2 µg/ml). In THP-1 cells, extracts were found to stimulate apoptosis caspase 3/7 activity. The IC50 values observed with osteosarcoma and leukaemia cells do not support a relevant cytotoxicity (as the figures abundantly exceeded 30 µg/ml), despites some selective activity exhibited with HL60 cells. The results confirm the validity of the extraction method, emphasizing the need to assess the selectivity of the interaction with cancer cells when screening for anticancer-drug candidates.

Brazilian Red Propolis: Extracts Production, Physicochemical Characterization, and Cytotoxicity Profile for Antitumor Activity.

Brazilian red propolis has been proposed as a new source of compounds with cytotoxic activity. Red propolis is a resinous material of vegetal origin, synthesized from the bees of the Appis mellifera family, with recognized biological properties. To obtain actives of low polarity and high cytotoxic profile from red propolis, in this work, we proposed a new solvent accelerated extraction method. A complete 23 factorial design was carried out to evaluate the influence of the independent variables or factors (e.g., temperature, number of cycles, and extraction time) on the dependent variable or response (i.e., yield of production). The extracts were analyzed by gas chromatography coupled with mass spectrometry for the identification of chemical compounds. Gas chromatography analysis revealed the presence of hydrocarbons, alcohols, ketones, ethers, and terpenes, such as lupeol, lupenone, and lupeol acetate, in most of the obtained extracts. To evaluate the cytotoxicity profile of the obtained bioactives, the 3-(4,5-dimethyl-2-thiazole)-2,5-diphenyl-2-H-tetrazolium bromide colorimetric assay was performed in different tumor cell lines (HCT116 and PC3). The results show that the extract obtained from 70 °C and one cycle of extraction of 10 min exhibited the highest cytotoxic activity against the tested cell lines. The highest yield, however, did not indicate the highest cytotoxic activity, but the optimal extraction conditions were indeed dependent on the temperature (i.e., 70 °C).

Phytochemical screening and in vitro antibacterial, antifungal, antioxidant and antitumor activities of the red propolis Alagoas / Triagem fitoquimico e atividade antibacteriana, antifúngica, antioxidante e antitumoral in vitro da propólis vermelha de Alagoas

Abstract The study aimed to evaluate the antimicrobial activity, antioxidant, toxicity and phytochemical screening of the Red Propolis Alagoas. Antimicrobial activity was evaluated by disk diffusion method. Determination of antioxidant activity was performed using the DPPH assay (1.1-diphenyl-2-picrylhydrazyl), FTC (ferric thiocyanate) and determination of phenolic compounds by Follin method. Toxicity was performed by the method of Artemia salina and cytotoxicity by MTT method. The phytochemical screening for the detection of allelochemicals was performed. The ethanol extract of propolis of Alagoas showed significant results for antimicrobial activity, and inhibitory activity for Staphylococcus aureus and Candida krusei. The antioxidant activity of the FTC method was 80% to 108.3% hydrogen peroxide kidnapping, the DPPH method showed an EC50 3.97 mg/mL, the content of total phenolic compounds was determined by calibration curve gallic acid, resulting from 0.0005 mg/100 g of gallic acid equivalent. The extract was non-toxic by A. salina method. The propolis extract showed high activity with a higher percentage than 75% inhibition of tumor cells OVCAR-8, SF-295 and HCT116. Chemical constituents were observed as flavonones, xanthones, flavonols, and Chalcones Auronas, Catechins and leucoanthocyanidins. It is concluded that the extract can be tested is considered a potential source of bioactive metabolites.

Resumo O trabalho teve como objetivo avaliar a atividade antimicrobiana, antioxidante, a toxicidade e a prospecção fitoquímica da Própolis Vermelha de Alagoas. A atividade antimicrobiana foi avaliada pelo método de difusão em disco. A determinação do potencial antioxidante foi realizada utilizando o método de DPPH (1,1-difenil-2-picrilhidrazil), FTC (Tiocianato Férrico) e determinação de compostos fenólicos pelo método de Follin. A toxicidade foi realizada pelo método de Artemia salina e a citotoxicidade pelo método do MTT. Foi realizada a prospecção fitoquímica para a pesquisa de aleloquímicos. O extrato etanólico da própolis vermelha de Alagoas apresentou resultados significantes para atividade antimicrobiana, tendo a atividade inibitória para Staphylococcus aureus e Candida krusei. Quanto a atividade antioxidante o método de FTC teve 80% a 108,3% de sequestro de peróxido de hidrogênio, o método de DPPH apresentou um CE50 de 3,97 μg/mL, o teor de compostos fenólicos totais foi determinado mediante curva de calibração do ácido gálico, tendo resultado de 0,0005 mg/100 g equivalente de ácido gálico. O extrato foi atóxico pelo método de A. salina. O extrato da própolis mostrou elevada atividade com percentual de inibição maior que 75% sobre células tumorais OVCAR-8, SF-295 e HCT116. Foram observados constituintes químicos como flavononas, xantonas, flavonóis, Chalconas e Auronas, Catequinas e Leucoantocianidinas. Conclui-se que o extrato testado pode ser considerado é uma fonte potencial de metabólitos bioativos.

Cornstarch-based wound dressing incorporated with hyaluronic acid and propolis: In vitro and in vivo studies.

The unique physicochemical and functional characteristics of starch-based biomaterials and wound dressings have been proposed for several biomedical applications. Film dressings of cornstarch/hyaluronic acid/ ethanolic extract of propolis (CS/HA/EEP) were prepared by solvent-casting and characterized by attenuated total reflectance/Fourier transform infrared spectroscopy, scanning electron microscopy, gas chromatography/mass spectrometry, light transmission, opacity measurements, EEP release, equilibrium swelling, and in vitro and in vivo evaluations. The CS/HA/0.5%EEP film dressing exhibited higher antibacterial activity against Staphylococcus aureus (2.08 ± 0.14 mm), Escherichia coli (2.64 ± 0.18 mm), and Staphylococcus epidermidis (1.02 ± 0.15 mm) in comparison with the CS, CS/HA, and CS/HA/0.25%EEP films. Also, it showed no cytotoxicity for the L929 fibroblast cells. This wound dressing could effectively accelerate the wound healing process at Wistar rats' skin excisions. These results indicate that enrichment of cornstarch wound dressings with HA and EEP can significantly enhance their potential efficacy as wound dressing material.

Phytochemical screening and in vitro antibacterial, antifungal, antioxidant and antitumor activities of the red propolis Alagoas.

The study aimed to evaluate the antimicrobial activity, antioxidant, toxicity and phytochemical screening of the Red Propolis Alagoas. Antimicrobial activity was evaluated by disk diffusion method. Determination of antioxidant activity was performed using the DPPH assay (1.1-diphenyl-2-picrylhydrazyl), FTC (ferric thiocyanate) and determination of phenolic compounds by Follin method. Toxicity was performed by the method of Artemia salina and cytotoxicity by MTT method. The phytochemical screening for the detection of allelochemicals was performed. The ethanol extract of propolis of Alagoas showed significant results for antimicrobial activity, and inhibitory activity for Staphylococcus aureus and Candida krusei. The antioxidant activity of the FTC method was 80% to 108.3% hydrogen peroxide kidnapping, the DPPH method showed an EC50 3.97 mg/mL, the content of total phenolic compounds was determined by calibration curve gallic acid, resulting from 0.0005 mg/100 g of gallic acid equivalent. The extract was non-toxic by A. salina method. The propolis extract showed high activity with a higher percentage than 75% inhibition of tumor cells OVCAR-8, SF-295 and HCT116. Chemical constituents were observed as flavonones, xanthones, flavonols, and Chalcones Auronas, Catechins and leucoanthocyanidins. It is concluded that the extract can be tested is considered a potential source of bioactive metabolites.

Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells.

The objective of this study was to evaluate and compare the cytotoxicity and genotoxicity on human fibroblast cell lines of sodium hypochlorite (NaOCl), chitosan and propolis as root canal irrigating solutions. Human fibroblast cells were exposed to chitosan, propolis and NaOCl for 4 and 24 h. Cell viability was assessed by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide, and oxidative DNA damage was assessed by determination of 8-hydroxydeoxyguanosine (8-OHdG) level with an ELISA kit. The data of cell cytotoxicity were analysed statistically using a test of one-way analysis of variance at a significance level of p < 0.05. In the NaOCI group, the 8-OHdG level was higher than in the chitosan group, but there was no statistical difference when compared with the other groups (p < 0.05). It was determined that the irrigation solutions were cytotoxic, depending on the dose and time. NaOCl was the most toxic solution after both 4 and 24 h of exposure (p < 0.05). Chitosan and propolis may be alternatives to NaOCl for irrigation solutions, because they are both less toxic and produce less oxidative DNA damage.

Engineering electrospun multicomponent polyurethane scaffolding platform comprising grapeseed oil and honey/propolis for bone tissue regeneration.

Essential oils play an important role in reducing the pain and inflammation caused by bone fracture.In this study, a scaffold was electrospun based on polyurethane (PU), grape seed oil, honey and propolis for bone tissue-engineering applications. The fiber diameter of the electrospun PU/grape seed oil scaffold and PU/grape seed oil/honey/propolis scaffold were observed to be reduced compared to the pristine PU control. FTIR analysis revealed the existence of grape seed oil, honey and propolis in PU identified by CH band peak shift and also hydrogen bond formation. The contact angle of PU/grape seed oil scaffold was found to increase owing to hydrophobic nature and the contact angle for the PU/grape seed/honey oil/propolis scaffold were decreased because of hydrophilic nature. Further, the prepared PU/grape seed oil and PU/grape seed oil/honey/propolis scaffold showed enhanced thermal stability and reduction in surface roughness than the control as revealed in thermogravimetric analysis (TGA) and atomic force microscopy (AFM) analysis. Further, the developed nanocomposite scaffold displayed delayed blood clotting time than the pristine PU in the activated prothrombin time (APTT) and partial thromboplastin time (PT) assay. The hemolytic assay and cytocompatibility studies revealed that the electrospun PU/grape seed oil and PU/grape seed oil/honey/propolis scaffold possess non-toxic behaviour to red blood cells (RBC) and human fibroblast cells (HDF) cells indicating better blood compatibility and cell viability rates. Hence, the newly developed electrospun nanofibrous composite scaffold with desirable characteristics might be used as an alternative candidate for bone tissue engineering applications.

Evaluation of radical scavenging activity, intestinal cell viability and antifungal activity of Brazilian propolis by-product.

Propolis is a natural adhesive resinous compound produced by honeybees to protect hives from bacteria and fungi, being extremely expensive for food industry. During propolis production, a resinous by-product is formed. This resinous waste is currently undervalued and underexploited. Accordingly, in this study the proximate physical and chemical quality, as well as the antioxidant activity, radical scavenging activity and cell viability of this by-product were evaluated and compared with propolis in order to boost new applications in food and pharmaceutical industries. The results revealed that the by-product meets the physical and chemical quality standards expected and showed that the propolis waste contains similar amounts of total phenolic content (TPC) and total flavonoid content (TFC) to propolis. Also, a good scavenging activity against reactive oxygen and nitrogen species (ROS and RNS, respectively) determined by the assays of superoxide anion radical (O2-), hydrogen peroxide (H2O2), hypochlorous acid (HOCl), nitric oxide (NO) and peroxyl radical (ROO) were determined. Linear positive correlations were established between the TPC of both samples and the antioxidant activity evaluated by three different methods (DPPH, ABTS and FRAP assays). The extracts were also screened for cell viability assays in two different intestinal cell lines (HT29-MTX and Caco-2), showing a viability concentration-dependent. Similarly, the Artemia salina assay, used to assess toxicity, demonstrated the concentration influence on results. Finally, the antifungal activity against ATCC species of Candida was demonstrated. These results suggest that propolis by-product can be used as a new rich source of bioactive compounds for different areas, such as food or pharmaceutical.

Mechanisms underlying the cytotoxic effect of propolis on human laryngeal epidermoid carcinoma cells.

Propolis has been used as a traditional remedy for centuries because of its beneficial effects, including anticancer properties. The aim of this study was to compare the cytotoxic mechanism of Cuban red propolis (CP) and Brazilian green propolis (BP) on human laryngeal carcinoma (HEp-2) cells. Cell viability, leakage of lactate dehydrogenase, fluorescence staining, mitochondrial membrane potential (ΔΨm) and the expression of pro/anti-apoptotic genes were assessed. Cell viability and cytotoxic assays suggested a dose-dependent effect of CP and BP extracts with a possible association of intracellular reactive oxygen species production and decreased ΔΨm. Both samples induced apoptosis via activation of TP53, CASP3, BAX, P21 signalling, and downregulation of BCL2 and BCL-XL. CP exerted a higher cytotoxic effect than BP extract. Our findings suggest further investigation of the main components of each propolis sample, what may lead to the development of strategies for the treatment of laryngeal cancer.

Assessment of toxicity and oxidative DNA damage of sodium hypochlorite, chitosan and propolis on fibroblast cells

Abstract The objective of this study was to evaluate and compare the cytotoxicity and genotoxicity on human fibroblast cell lines of sodium hypochlorite (NaOCl), chitosan and propolis as root canal irrigating solutions. Human fibroblast cells were exposed to chitosan, propolis and NaOCl for 4 and 24 h. Cell viability was assessed by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide, and oxidative DNA damage was assessed by determination of 8-hydroxydeoxyguanosine (8-OHdG) level with an ELISA kit. The data of cell cytotoxicity were analysed statistically using a test of one-way analysis of variance at a significance level of p < 0.05. In the NaOCI group, the 8-OHdG level was higher than in the chitosan group, but there was no statistical difference when compared with the other groups (p < 0.05). It was determined that the irrigation solutions were cytotoxic, depending on the dose and time. NaOCl was the most toxic solution after both 4 and 24 h of exposure (p < 0.05). Chitosan and propolis may be alternatives to NaOCl for irrigation solutions, because they are both less toxic and produce less oxidative DNA damage.

Antioxidant and cytotoxic activity of propolis of Plebeia droryana and Apis mellifera (Hymenoptera, Apidae) from the Brazilian Cerrado biome.

Propolis is a complex bioactive mixture produced by bees, known to have different biological activities, especially in countries where there is a rich biodiversity of plant species. The objective of this study was to determine the chemical composition and evaluate the antioxidant and cytotoxic properties of Brazilian propolis from the species Plebeia droryana and Apis mellifera found in Mato Grosso do Sul, Brazil. In the ethanolic extracts of P. droryana propolis (ExEP-P) and A. mellifera (ExEP-A) acids, phenolic compounds, terpenes and tocopherol were identified as major compounds. Both extracts presented antioxidant activity against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical, the maximum activities being 500 µg/mL (ExEP-P) and 300 µg/mL (ExEP-A). However, only ExEP-A was able to inhibit lipid peroxidation induced by the oxidizing agent 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), which inhibited oxidative hemolysis and reduced the levels of malondialdehyde (MDA) in human erythrocytes for 4 h of incubation. The extracts also reduced the cell viability of the K562 erythroleukemia tumour line, with a predominance of necrotic death. Thus, it is concluded that the propolis produced by P. droryana and A. mellifera contain important compounds capable of minimizing the action of oxidizing substances in the organism and reducing the viability of erythroleukemia cells.

Estudio de la actividad antioxidante y antitumoral del propóleo / Study on the antioxidant and antitumoral activity of propolis

Introducción: El propóleo es la sustancia que protege a la colmena, es una resina de composición compleja y viscosa que las abejas utilizan en la reparación y protección de la colmena. El material del que procede el propóleo son las resinas, brotes y pecíolos de las hojas de diferentes vegetales, por ello presenta una composición química muy compleja que varía en función de la flora de recolección de las abejas. Posee capacidad antimicrobiana, antiinflamatoria que está relacionada con su poder antioxidante, inmunomoduladora, entre otras. Objetivos: En este trabajo se estudian las actividades antioxidantes y antitumorales de propóleos de distintas zonas de la provincia de Málaga comparándolos con uno de la región de Bohemia al sur de la República Checa. Material y métodos: La actividad antioxidante se evaluó según el método ABTS+/S2O8K2. Además, se estimó la cantidad de proteínas totales a partir del contenido de nitrógeno y posteriormente se determinó la citotoxicidad y actividad antitumoral del propóleo del Puerto de la Torre, al norte de Málaga, según el método del bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazolio. Resultados: Se observó que el propóleo presenta una elevada actividad antioxidante, aunque tiene una menor cantidad de proteínas. El propóleo presenta elevada toxicidad y mayor actividad antitumoral frente al cáncer de colon que al de leucemia. Conclusión: Con todos estos datos obtenidos se puede concluir que el propóleo presenta diferentes actividades de interés para la industria alimentaria o cosmética, entre las que destaca su elevado poder antioxidante y su capacidad como antitumoral

Introduction: Propolis is the substance that protects the hive, a resin of complex and viscous composition bees use in the repair and protection of the hive. The material from which propolis arises are the resins, shoots and petioles of the leaves of different plants, so it has a very complex chemical composition that varies depending on the flora of the bees collection. It offers an antimicrobial, anti-inflammatory capacity related to its antioxidant, immunomodulatory power, among others. Aims: In this work, antioxidant and antitumoral activities of different propolis collected from different areas of the province of Malaga, comparing them with one from the Bohemian region to the south of the Czech Republic are studied. Material and methods: Antioxidant activity was determined according to the ABTS+/S2O8K2 method. In addition, the quantity of total proteins from the nitrogen content and subsequently the cytotoxicity and antitumoral activity of the propolis of Puerto de la Torre, north of Malaga, are measured according to the 3-(4,5-dimetiltiazol-2-il)-2,5-diphenyl tetrazolium bromide method. Results: It was observed that propolis has high antioxidant activity, although it has a lower amount of proteins. Propolis has high toxicity and higher antitumoral activity against colon cancer than leukemia. Discussion: With all these data, it can be concluded that propolis offers different activities of interest, for the food and cosmetic industry, among which the high antioxidant and antitumoral capacity

Antioxidant, Cytotoxic, and Toxic Activities of Propolis from Two Native Bees in Brazil: Scaptotrigona depilis and Melipona quadrifasciata anthidioides.

Propolis is a natural mixture of compounds produced by various bee species, including stingless bees. This compound has been shown to exhibit antioxidant, antiproliferative, and antitumor activities. The present study aimed to determine the chemical constituents as well as the antioxidant, cytotoxic, and toxic activities of ethanol extracts of propolis obtained from the stingless bees Scaptotrigona depilis and Melipona quadrifasciata anthidioides, which are found in Brazil. Phytosterols, terpenes, phenolic compounds, and tocopherol were identified in the ethanol extracts of propolis (EEPs) in different concentrations. The compounds stigmasterol, taraxasterol, vanilic acid, caffeic acid, quercetin, luteolin, and apigenin were found only in EEP-M. The EEPs were able to scavenge the free radicals 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and protected human erythrocytes against lipid peroxidation, with the latter effect being demonstrated by their antihemolytic activity and inhibition of malondialdehyde formation. The EEPs showed cytotoxic activity against erythroleukemic cells and necrosis was the main mechanism of death observed. In addition, the concentrations at which the EEPs were cytotoxic were not toxic against Caenorhabditis elegans. In this context, it is concluded that EEP-S and EEP-M show antioxidant and cytotoxic activities and are promising bioactive mixtures for the control of diseases associated with oxidative stress and tumor cell proliferation.

Chemical and cytotoxic analyses of brown Brazilian propolis (Apis mellifera) and its in vitro activity against itraconazole-resistant Sporothrix brasiliensis.

This study aimed to evaluate the chemical composition and cytotoxic activity of brown Brazilian propolis and its in vitro activity against itraconazole-resistant Sporothrix brasiliensis from animal sporotrichosis. Propolis was acquired commercially and prepared as a hydroalcoholic extract. Chemical analysis was evaluated by liquid chromatography coupled to mass spectrometry of ultra-efficiency. The cell viability was evaluated by MTT test in MDBK cells of 50 to 0.09 µg/mL. For antifungal tests, twenty isolates of Sporothrix brasiliensis from dogs (n = 11) and cats (n = 9) with sporotrichosis were tested to itraconazole (16-0.0313 µg/mL) and to propolis (3.125-0.09 mg/mL) by broth microdilution technique (CLSI M38-A2), adapted to natural products. The results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). Itraconazole showed activity between MIC values of 0.25 to greater than 16 µg/mL, and 88.9% (08/09) and 72.7% (08/11) of S. brasiliensis from cats and dogs, respectively, were considered itraconazole-resistant. All Sporothrix brasiliensis were sensitive to brown propolis between MIC values of 0.19-1.56 mg/mL, including the itraconazole-resistant isolates, whereas the MFC values of propolis were from 0.78 to greater than 3.125 mg/mL. Propolis maintained a medium to high cell viability between concentration of 0.78 to 0.09 µg/mL, and p-coumaric acid was the major compound. Brown Brazilian propolis is a promising antifungal candidate against sporotrichosis and more studies need to be undertaken to evaluate its safe use to understand its efficacy.

Avaliação in-vitro da atividade antifúngica e da citotoxicidade da própolis de copaíba contra espécies do gênero Candida spp / In-vitro evaluation of the antifungal activity and cytotoxicity of copaiba propolis against Candida spp

OBJETIVOS: verificar a atividade antifúngica e a citotoxicidade da própolis de copaíba e de um protótipo de enxaguante bucal contra espécies do gênero Candida spp. METODOLOGIA: o perfil cromatográfico do extrato de própolis foi realizado por Cromatografia Líquida de Ultraeficiência em Fase Reversa (RP-UPLC). A atividade antifúngica do extrato de própolis de copaíba (EPC) e do seu protótipo de enxaguante bucal contra C. albicans, C. tropicalis e C. krusei foi avaliada por microdiluição em caldo e discodifusão em ágar. A citotoxicidade foi verificada pelo ensaio de MTT em fibroblastos 3T3 ¿ L1. RESULTADOS: Fenóis foram os principais compostos encontrados, sendo identificados dentre eles o ácido caféico (ácido fenólico) e canferol (flavonóide). Os valores de CIM foram 156 ug/ml, 312 ug/ml e 625 ug/ml para C. albicans, C. tropicalis e C. krusei respectivamente, indicando atividade antifúngica presente, mas moderada. Os valores das zonas de inibição verificaram inibição do crescimento das leveduras e sugeriram uma atividade fungistática do enxaguante bucal. Em relação aos testes de citotoxicidade, o EPC exerce, em baixas concentrações, um efeito dose dependente sobre a proliferação de fibroblastos 3T3-L1 e pode exercer um efeito regenerador de tecidos. CONCLUSÕES: a própolis investigada apresenta potencial antifúngico moderado contra Candida spp., podendo ser o enxaguante bucal uma opção terapêutica no combate da candidíase oral, já que o seu tratamento é baseado no controle e não na erradicação das leveduras. Estudos futuros sobre a caracterização dos seus compostos químicos, aperfeiçoamento do produto e ensaios clínicos deverão ser realizados

Objectives: verify the antifungal activity and cytotoxicity of copaiba propolis and its mouthwash prototype against Candida spp. Methodology: the chromatographic profile of the propolis extract was perpormed by Reverse Phase Ultra-efficiency Liquid Chromatography (RP-UPLC)....

Immunomodulatory/inflammatory effects of geopropolis produced by Melipona fasciculata Smith in combination with doxorubicin on THP-1 cells.

OBJECTIVES: Geopropolis (GEO) in combination with doxorubicin (DOX) reduced HEp-2 cells viability compared to GEO and DOX alone. A possible effect of this combination on the innate immunity could take place, and its effects were analysed on THP-1 cell - a human leukaemia monocytic cell line used as a model to study monocyte activity and macrophage activity, assessing cell viability, expression of cell markers and cytokine production. METHODS: THP-1 cells were incubated with GEO, DOX and their combination. Cell viability was assessed by MTT assay, cell markers expression by flow cytometry and cytokine production by ELISA. KEY FINDINGS: GEO + DOX did not affect cell viability. GEO alone or in combination increased TLR-4 and CD80 but not HLA-DR and TLR-2 expression. GEO stimulated TNF-α production while DOX alone or in combination did not affect it. GEO alone or in combination inhibited IL-6 production. CONCLUSIONS: GEO exerted a pro-inflammatory profile by increasing TLR-4 and CD80 expression and TNF-α production, favouring the activation of the immune/inflammatory response. GEO + DOX did not affect cell viability and presented an immunomodulatory action. Lower concentrations of DOX combined to GEO could be used in cancer patients, avoiding side effects and benefiting from the biological properties of GEO.

In vivo evaluation of mutagenic and recombinagenic activities of Brazilian propolis.

Propolis is a resinous, complex mixture of compounds collected by the bee species Apis mellifera. This study investigated the genotoxicity of green and brown propolis collected in southeast Brazil using the somatic mutation and recombination test (SMART) in Drosophila melanogaster. The effect of five concentrations (0.5, 1.0, 2.0, 4.0, and 7.5 mg/mL) of both propolis types was analyzed in standard (ST) and high-bioactivation (HB) crosses, which have normal and high levels of cytochrome P450 enzymes, respectively. The results show that the types of propolis evaluated have no mutagenic action, in either cross. Moreover, chromatography findings revealed that the propolis types analyzed have different chemical compositions. Brown propolis had lower levels of polyphenols (∼7.2 mg/mL), compared to the green type (34.9 mg/g). Taken together, the findings of the present study and literature reports point to the safety in consuming low amounts of propolis, considering the risk of genetic damage, and confirm the absence of mutagenic and recombinagenic actions of the propolis types investigated.