Artemisia vulgaris Induces Tumor-Selective Ferroptosis and Necroptosis via Lysosomal Ca2+ Signaling.

OBJECTIVE: To evaluate the chemical composition and effects of Artemisia vulgaris (AV) hydroalcoholic extract (HEAV) on breast cancer cells (MCF-7 and SKBR-3), chronic myeloid leukemia (K562) and NIH/3T3 fibroblasts. METHODS: Phytochemical analysis of HEAV was done by high-performance liquid chromatography-mass (HPLC) spectrometry. Viability and cell death studies were performed using trypan blue and Annexin/FITC-7AAD, respectively. Ferrostatin-1 (Fer-1) and necrostatin-1 (Nec-1) were used to assess the mode of HEAV-induced cell death and acetoxymethylester (BAPTA-AM) was used to verify the involvement of cytosolic calcium in this event. Cytosolic calcium measurements were made using Fura-2-AM. RESULTS: HEAV decreased the viability of MCF-7, SKBR-3 and K562 cells (P<0.05). The viability of HEAV-treated K562 cells was reduced compared to HEAV-exposed fibroblasts (P<0.05). Treatment of K562 cells with HEAV induced cell death primarily by late apoptosis and necrosis in assays using annexin V-FITC/7-AAD (P<0.05). The use of Nec-1 and Fer-1 increased the viability of K562 cells treated with HEAV relative to cells exposed to HEAV alone (P<0.01). HEAV-induced Ca2+ release mainly from lysosomes in K562 cells (P<0.01). Furthermore, BAPTA-AM, an intracellular Ca2+ chelator, decreased the number of non-viable cells treated with HEAV (P<0.05). CONCLUSIONS: HEAV is cytotoxic and activates several modalities of cell death, which are partially dependent on lysosomal release of Ca2+. These effects may be related to artemisinin and caffeoylquinic acids, the main compounds identified in HEAV.

Synergistic Antibiofilm Action of Cinnamomum verum and Brazilian Green Propolis Hydroethanolic Extracts against Multidrug-Resistant Strains of Acinetobacter baumannii and Pseudomonas aeruginosa and Their Biocompatibility on Human Keratinocytes.

The accumulated dental biofilm can be a source of oral bacteria that are aspirated into the lower respiratory tract causing ventilator-associated pneumonia in hospitalized patients. The aim of this study was to evaluate the synergistic antibiofilm action of the produced and phytochemically characterized extracts of Cinnamomum verum and Brazilian green propolis (BGP) hydroethanolic extracts against multidrug-resistant clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition to their biocompatibility on human keratinocyte cell lines (HaCaT). For this, High-performance liquid chromatography analysis of the plant extracts was performed; then the minimum inhibitory and minimum bactericidal concentrations of the extracts were determined; and antibiofilm activity was evaluated with MTT assay to prevent biofilm formation and to reduce the mature biofilms. The cytotoxicity of the extracts was verified using the MTT colorimetric test, evaluating the cellular enzymatic activity. The data were analyzed with one-way ANOVA and Tukey's tests as well as Kruskal-Wallis and Dunn's tests, considering a significance level of 5%. It was possible to identify the cinnamic aldehyde in C. verum and p-coumaric, caffeic, and caffeoylquinic acids as well as flavonoids such as kaempferol and kaempferide and Artepillin-C in BGP. The combined extracts were effective in preventing biofilm formation and reducing the mature biofilms of A. baumannii and P. aeruginosa. Moreover, both extracts were biocompatible in different concentrations. Therefore, C. verum and BGP hydroethanolic extracts have bactericidal and antibiofilm action against multidrug resistant strains of A. baumannii and P. aeruginosa. In addition, the combined extracts were capable of expressively inhibiting the formation of A. baumannii and P. aeruginosa biofilms (prophylactic effect) acting similarly to 0.12% chlorhexidine gluconate.

Antifungal Action of Herbal Plants' Glycolic Extracts against Candida Species.

Candida spp. cause fungal infection that affects patients' oral health. This study aimed to evaluate the isolated and synergistic antifungal effect of Rosa centifolia L., Curcuma longa L., Rosmarinus officinalis L., and Punica granatum L. glycolic extracts against Candida albicans, Candida dubliniensis, Candida tropicalis, and Candida krusei planktonic and biofilm forms. The plant extracts were chemically characterized and the main compounds were quantified by high-performance liquid chromatography (HPLC-DAD) analysis. The minimum inhibitory and minimum fungicidal concentrations of the extracts were determined, and antibiofilm activity was evaluated by MTT assay. Data were analyzed by one-way ANOVA and Tukey's tests, and by Kruskal-Wallis and Dunn's tests, considering a significance level of 5%. The main compounds identified in each of the extracts were: p-coumaric acid (2153.22 µg/100 mL) in the rosemary extract, gallotannins (4318.31 µg/100 mL) in the pomegranate extract, quercetin derivatives (3316.50 µg/100 mL) in the extract of white roses, and curcumin (135.09 µg/100 mL) in the turmeric extract. The combination of R. centifolia and C. longa glycolic extracts was effective against C. albicans, C. dubliniensis, and C. tropicalis biofilms over different periods (p < 0.05). The combination of R. officinalis and P. granatum glycolic extracts was effective against C. albicans and C. krusei biofilms after 30 min, and against C. tropicalis after 24 h, with all combinations showing an average reduction of 50% in cell viability (p < 0.05). In conclusion, the combined plant extracts have antifungal and antibiofilm action against Candida spp. in different concentrations and times of action.

Effects of Calendula officinalis and Capsicum annum glycolic extracts on planktonic cells and biofilms of multidrug-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa.

Empirical knowledge of natural plant extracts is increasingly proving to be a promising field. The effect of Calendula officinalis L. (CO) and Capsicum annum (CA) glycolic extracts (GlExt) have potential that should be further developed in microbial tests. The effect of CO-GlExt and CA-GlExt was evaluated on eight multidrug-resistant clinical strains of Klebsiella pneumoniae and Pseudomonas aeruginosa, as well as collection strains for each bacterial. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined in comparison with 0.12% chlorhexidine. The tests were performed on single species biofilms, at 5 min and 24 h, using the MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. The MIC and MBC of the extract ranged from 1.56 to 50 mg mL-1 for all strains evaluated. Analysis of the MTT assay revealed a strong antimicrobial potential of CA-GlExt, comparable to chlorhexidine. The findings suggest that CA-GlExt is effective against multidrug-resistant strains of K. pneumoniae and P. aeruginosa in planktonic state and biofilms.

Molecular Dereplication and In Vitro and In Silico Pharmacological Evaluation of Coriandrum sativum against Neuroblastoma Cells.

The aim of this study was to investigate the cytotoxic activity of the Coriandrum sativum (C. sativum) ethanolic extract (CSEE) in neuroblastoma cells, chemically characterize the compounds present in the CSEE, and predict the molecular interactions and properties of ADME. Thus, after obtaining the CSEE and performing its chemical characterization through dereplication methods using UPLC/DAD-ESI/HRMS/MS, PM6 methods and the SwissADME drug design platform were used in order to predict molecular interactions and ADME properties. The CSEE was tested for 24 h in neuroblastoma cells to the establishment of the IC50 dose. Then, the cell death was evaluated, using annexin-PI, as well as the activity of the effector caspase 3, and the protein and mRNA levels of Bax and Bcl-2 were analyzed by ELISA and RT-PCR, respectively. By UHPLC/DAD/HRMS-MS/MS analysis, the CSEE showed a high content of isocoumarins-dihydrocoriandrin, coriandrin, and coriandrones A and B, as well as nitrogenated compounds (adenine, adenosine, and tryptophan). Flavonoids (apigenin, hyperoside, and rutin), phospholipids (PAF C-16 and LysoPC (16:0)), and acylglicerol were also identified in lower amount as important compounds with antioxidant activity. The in silico approach results showed that the compounds 1 to 6, which are found mostly in the C. sativum extract, obey the "Five Rules" of Lipinski, suggesting a good pharmacokinetic activity of these compounds when administered orally. The IC50 dose of CSEE (20 µg/mL) inhibited cell proliferation and promoted cell death by the accumulation of cleaved caspase-3 and the externalization of phosphatidylserine. Furthermore, CSEE decreased Bcl-2 and increased Bax, both protein and mRNA levels, suggesting an apoptotic mechanism. CSEE presents cytotoxic effects, promoting cell death. In addition to the promising results predicted through the in silico approach for all compounds, the compound 6 showed the best results in relation to stability due to its GAP value.

The biocompatibility and antifungal effect of Rosmarinus officinalis against Candida albicans in Galleria mellonella model.

This study was performed to evaluate the biocompatibility and antifungal effect of Rosmarinus officinalis against Candida albicans in Galleria mellonella model. Five different concentrations of R. officinalis glycolic extract (50; 25; 12.5 e 6.25 mg/mL) were used to evaluate its biocompatibility in G. mellonella model, in which the nystatin suspension (100; 50; 25; 12.5 e 6.25%) was used as a control group. The antifungal action of R. officinalis glycolic extract was evaluated on C. albicans for 72, 48 and 12 h at two different phases: (1) using the extract as therapeutic agent; and (2) using the extract as prophylactic agent. PBS was used as a negative control group. G. mellonella survival curves were plotted using the Kaplan-Meier method and statistical analysis was performed using the log-rank test (Mantel-Cox) and the significance level was set at (α ≤ 0.05). There was no significant difference among the groups in which all were biocompatible except of a significant death rate of 26.6% with nystatin 100%. In phase 1, it was found that after 7 days, there was no statistically significant difference among the prophylactic treatment groups. In phase 2, the groups of R. officinalis 6.25 mg/mL for 72 h and R. officinalis of 12.5 mg/mL for 24 h promoted the survival rate of the larvae in comparison with the control group with a significant difference (p = 0.017) and (p = 0.032) respectively. Therefore, R. officinalis extract is biocompatible in different concentrations and can be used as a prophylactic agent against fungal infection.

Curcuma longa L. Extract and Photodynamic Therapy are Effective against Candida spp. and Do Not Show Toxicity In Vivo.

Radiotherapy induces a higher level of Candida spp. colonization, resulting in oral candidiasis. This study aimed to evaluate the phototransformation potential of the glycolic extract of Curcuma longa (C. longa); the antifungal activity of C. longa, curcumin, and antifungal photodynamic therapy (aPDT) with blue light-emitting diodes "LED" on Candida albicans and Candida tropicalis in vitro; and the toxicity of C. longa and curcumin in Galleria mellonella model. In order to confirm the light absorption capacity of the C. longa extract, its phototransformation potential was evaluated. The antifungal effect of C. longa, curcumin, and aPDT was evaluated over Candida spp. Finally, the toxicity of C. longa and curcumin was evaluated on the Galleria mellonella model. The data were analyzed using the GraphPad Prism 5.0 software considering α = 5%. It was found that C. longa, curcumin, and aPDT using blue LED have an antifungal effect over C. albicans and C. tropicalis. The extract of C. longa 100 mg/mL and curcumin 200 µg/mL do not show toxicity on Galleria mellonella model.

Influence of Dentin Priming with Tannin-Rich Plant Extracts on the Longevity of Bonded Composite Restorations.

OBJECTIVE: This in vitro study evaluated the influence of bioactive plant extracts as dentin biomodifying agents to improve the longevity of bonded restorations. For that, plant extracts were applied to the dentin surface prior to the adhesive system. MATERIALS AND METHODS: Bovine incisors were ground flat to obtain 2 mm thick slices in which conical preparations were made (N = 10). Tannin-containing plant extracts were applied to dentin before the application of the restorative system, as follows: control group (untreated, CTL), chlorhexidine 0.12% (CHX), mastruz (Dysphania ambrosioides, MTZ), cat's claw (Uncaria tomentosa, CTC), guarana (Paullinia cupana, GUA), galla chinensis (Rhus chinensis, GCH), and tannic acid (extracted from Acacia decurrens, TNA). The push-out bond strength test was conducted (0.5 mm/min). Dentin biomodification was assessed by the modulus of elasticity and mass change in bovine tooth sections (0.5 × 1.7 × 7.0 mm). The dentin staining after extract treatments of dentin slices was compared. The dentin surface wettability was also evaluated by means of the contact angles of the adhesive system with the dentin surface and compared with the untreated control group. Data were subjected to ANOVA/Tukey's test (α = 0.05). RESULTS: The bond strength of the restoratives to dentin was not significantly improved by the plant extracts, irrespective of the evaluation time (p > 0.05). Except for TNA, the elastic modulus of demineralized dentin significantly reduced after treatment with the plant extracts (p < 0.05). The dentin staining correlated with the tannin content of the extracts. The contact angle was significantly reduced when treated with CTC, GCH, and TNA. CONCLUSIONS: The tannin-containing extracts had a questionable effect on the longevity of bonded restorations. The dentin modulus was negatively affected by the extract treatments. Although some of the extracts changed the contact angle, which seems to improve the adhesive monomer permeation, the tannin-rich plant extract application prior to adhesive application was proven to be clinically unfeasible due to dentin staining.

Antibacterial Efficacy of Triple Antibiotic Medication With Macrogol (3Mix-MP), Traditional Triple Antibiotic Paste, Calcium Hydroxide, and Ethanol Extract of Propolis: An Intratubular Dentin Ex Vivo Confocal Laser Scanning Microscopic Study.

INTRODUCTION: This study aimed to assess the effectiveness of antibacterial activity of medications used in regenerative endodontic treatment. METHODS: Sixty-seven dentin cylinders of single-rooted teeth were contaminated with a culture of Enterococcus faecalis (ATCC 29212; American Type Culture Collection, Manassas, VA) for 5 days. Samples were divided into 1 control group and the following experimental groups according to the medication applied: traditional triple antibiotic paste (TAP), clindamycin-modified TAP (mTAP), triple antibiotic medication with macrogol (3Mix-MP), clindamycin-modified 3Mix-MP (m3Mix-MP), calcium hydroxide (CH), and ethanol extract of propolis (EEP). After 14 days, the medications were removed, and the samples were submitted to confocal laser scanning microscopic analysis to quantify the percentage of viable bacteria. The distribution of data was confirmed by the Shapiro-Wilk test. The Kruskal-Wallis and Dunn tests were used for intergroup comparisons, and the Wilcoxon test was used for comparison between superficial and deep antibacterial efficacy for the same medication. The level of significance was set at P < .05. RESULTS: 3Mix-MP and m3Mix-MP presented significantly higher antibacterial efficacy compared with the other tested medications (P < .05), except for mTAP. mTAP was more effective than TAP (P < .05). The antibacterial efficacy of EEP and CH did not differ significantly from TAP and mTAP (P > .05). All medications showed effective antibacterial action compared with the control group (P < .05). CONCLUSIONS: 3Mix-MP and m3Mix-MP, which present extremely high concentrations of antibiotics (1500 mg/mL), were not more effective than mTAP at the concentration recommended by the American Association of Endodontists (5 mg/mL). Moreover, CH and EEP were as effective as TAP and mTAP.

Pimentas do gênero Capsicum: propriedades e ações terapéuticas / Pimientos del género Capsicum: propiedades y acciones terapéuticas / Capsicum peppers: properties and therapeutic actions

As especies do genero Capsicum L. vem sendo estudadas por pesquisadores do mundo inteiro. A pungencia e o atributo principal das pimentas e as substancias responséveis por esta ardencia sao denominadas de capsaicinoides. O mais estudado ea capsaicina, sua rota biossintetica e atraves da via dos fenilpropanoides e écidos graxos. A capsaicina e um agonista exogeno do receptor TRPV1(transient receptor potential vanilloid type-7). O TRPV1 contem uma subunidade sensivel ao calor responsével pela sensaçao de queimadura causada pela capsaicina. Quando aplicada na pele, promove uma resposta analgesica devido a dessensibilizaçao dos neuronios sensoriais causados pelo esgotamento da substancia P. A meia vida da capsaicina e vinte e quatro horas quando utilizada por via oral. Sua concentraçao maxima atinge o figado, rins e intestino ern uma hora apos administraçao oral. A capsaicina e principalmente eliminada pelos rins, com uma pequena proporçao nao transformada excretada nas fezes e na urina. Na aplicagao topica, a biotransformaçao da capsaicina foi considerada lenta e a maior parte da mesma permaneceu inalterada. A capsaicina e seus anélogos tem sido utilizados em cremes e patches para tratar sindromes de dor cronica como neuralgia pos-herpetica, dores musculoesqueleticas, neuropatia diabetica, osteoartrite e artrite reumatoide. A capsaicina tambem tem atividade antihiperlipidemica, propiedades anti-inflamatorias, antioxidantes e e efetiva no tratamento da dor associada com artrite e cistite. O capsiato, presente nas pimentas vermelhas nao pungentes tambem estimula o receptor TRPV1, sendo capaz de aumentar o metabolismo por estimulagao do sistema nervoso simpético, alem de ser provido de agao antiinflamatoria, porem por mecanismo ainda desconhecido (AU)

Las especies del género Capsicum, han sido estudiadas por los investigadores de todo el mundo. La pungencia es el principal atributo de los pimientos picantes y las sustancias responsables son los capsaicinoides. La capsaicina es la mas estudiada, su ruta biosintética es a través de la vía de fenilpropanoides y ácidos grasos. La capsaicina es un receptor TRPV1 agonista exógeno (receptor de potencial transitorio vaniloide tipo 1). El TRPV1 contiene una subunidad sensible al calor responsable de la sensación de ardor causada por la capsaicina. Cuando se aplica a la piel se promueve una respuesta analgésica debido a la desensibilización de las neuronas sensoriales causadas por el agotamiento de la sustancia P. La Vida media de la capsaicina es de 24 horas cuando se usa por via oral. Su distribución máxima alcanza el hígado, el riñón y el intestino en una hora después de la administración oral. La capsaicina se elimina principalmente por via renal, con una pequefia proporción no transformada excretada en las heces y en la orina. En la aplicación tópica de capsaicina la biotransformación se consideró lenta, y la mayor parte de la misma permaneció inalterada. La capsaicina y sus análogos han sido utilizados en cremas y apósitos para el tratamiento de síndromes de dolor crónico, tales como neuralgia postherpética, dolor musculo esquelético, neuropatía diabética, la osteoartritis y artritis reumatoide. La capsaicina también tiene actividad hipolipemiante, antiinflamatoria, antioxidante y es eficaz en el tratamiento del dolor asociado a artritis y cistitis. El capsiato, presente en pimientos rojos no picantes, también estimula el receptor TRPV1, siendo capaz de aumentar el metabolismo mediante la estimulación del sistema nervioso simpático, y presentando, además, acción antiinflamatoria, por un mecanismo aun desconocido (AU)

The species of the genus Capsicum have been studied by researchers worldwide. Pungency is the main attribute of peppers and it is due to the capsaicinoids. The most studied is capsaicin, biosynthetised through the phenylpropanoid and fatty acid pathway. Capsaicin is an exogenous agonist of TRPV1 receptor (transient receptor potential vanilloid type-1). The TRPV1 contains a subunit sensitive to heat, responsible for the burning sensation caused by capsaicin. When applied to the skin it promotes an analgesic response due to desensitization of sensory neurons caused by the depletion of substance P. The half-life of capsaicin is twenty-four hours when used orally. Its maximum concentration reaches the liver, kidney and intestine one hour after oral administration. Capsaicin is eliminated primarily by kidneys, with a small proportion of untransformed excreted in faeces and urine. After topical application, the biotransformation of capsaicin was considered slow, and it was mostly unchanged. Capsaicin and its analogues have been used in creams and patches to treat chronic pain syndromes, such as postherpetic neuralgia, musculoskeletal pain, diabetic neuropathy, osteoarthritis and rheumatoid arthritis. Capsaicin also has antihyperlipidemic, anti-inflammatory and antioxidant activities, being effective in the treatment of pain associated with arthritis and cystitis. The capsiate, present in non-pungent red peppers, also stimulates the TRPV1 receptor, being able to increase metabolism by stimulating the sympathetic nervous system, and showing, in addition, anti-inflammatory activity by a mechanism still unknown (AU)

Potential utility of hyperbaric oxygen therapy and propolis in enhancing the leishmanicidal activity of glucantime.

In this study we investigated the efficacy of hyperbaric oxygen (HBO) therapy, alone or combined with the pentavalent antimonial glucantime on Leishmania amazonensis infection. In parallel, the effect of Brazilian red propolis gel (propain) alone or combined with glucantime on L. amazonensis infection was evaluated. The inhibition of the infection in macrophages treated with glucantime in combination with HBO exposition was greater than that of macrophages treated with glucantime alone or HBO alone. The susceptible mouse strain BALB/c infected in the shaved rump with L. amazonensis treated with glucantime and exposed to HBO showed: time points in the course of the disease in which lesions were smaller than those of mice treated with glucantime alone and revascularization of the skin in the lesion site; interferon-gamma (IFN-g) levels were not elevated in lymph node cells from these animals. Propain alone was not efficient against lesions, although less exudative lesions were observed in animals treated with propain alone or combined with glucantime. These results reveal the potential value of HBO and red propolis in combination with glucantime for treating cutaneous leishmaniasis and encourage further studies on the effect of more aggressive HBO, propolis and glucantime therapies on different mouse models of leishmaniasis.

Potential utility of hyperbaric oxygen therapy and propolis in enhancing the leishmanicidal activity of glucantime / A utilidade da terapia de oxigenação hiperbárica e própolis em potencializar a atividade leishmanicida do glucantime

In this study we investigated the efficacy of hyperbaric oxygen (HBO) therapy, alone or combined with the pentavalent antimonial glucantime on Leishmania amazonensis infection. In parallel, the effect of Brazilian red propolis gel (propain) alone or combined with glucantime on L. amazonensis infection was evaluated. The inhibition of the infection in macrophages treated with glucantime in combination with HBO exposition was greater than that of macrophages treated with glucantime alone or HBO alone. The susceptible mouse strain BALB/c infected in the shaved rump with L. amazonensis treated with glucantime and exposed to HBO showed: time points in the course of the disease in which lesions were smaller than those of mice treated with glucantime alone and revascularization of the skin in the lesion site; interferon-gamma (IFN-g) levels were not elevated in lymph node cells from these animals. Propain alone was not efficient against lesions, although less exudative lesions were observed in animals treated with propain alone or combined with glucantime. These results reveal the potential value of HBO and red propolis in combination with glucantime for treating cutaneous leishmaniasis and encourage further studies on the effect of more aggressive HBO, propolis and glucantime therapies on different mouse models of leishmaniasis.

Nesse trabalho foi avaliada a eficácia da terapia da oxigenação hiperbárica (HBO), aplicada em combinação ou não com o tratamento com glucantime, durante a infecção com Leishmania amazonensis. O efeito de gel da própolis vermelha de origem brasileira (propaina) aplicado em combinação ou não com o tratamento com glucantime, também foi avaliado durante infecção com esse parasita. A inibição da infecção de macrófagos tratados com glucantime em combinação com HBO foi maior que a de macrófagos tratados apenas com glucantime ou HBO. A linhagem murina susceptível, BALB/c, infectada no dorso com L. amazonensis, tratada com glucantime e exposta a HBO, mostrou durante o curso da doença, fases em que as lesões eram menores do que a de camundongos apenas tratados com glucantime; observou-se revascularização da pele da lesão e baixa produção de interferon-gama em células de linfonodos desses animais. O tratamento com propaina não foi eficiente na cura das lesões, apesar de lesões menos exsudativas serem observadas em animais tratados com propaina ou propaina combinada ao tratamento com glucantime. Os resultados demonstram que tanto HBO como a própolis vermelha em combinação com glucantime, são promissoras no tratamento da leishmaniose cutânea. Novos estudos devem ser realizados para avaliar tratamentos e outros protocolos em diferentes modelos murinos da leishmaniose.

Evaluation of the in vitro antimicrobial activity of an ethanol extract of Brazilian classified propolis on strains of Staphylococcus aureus

Staphylococcus aureus (S. aureus) is one of the most frequent causes of hospital acquired infections. With the increase in multiple drug resistant strains, natural products such as propolis are a stratagem for new product discovery. The aims of this study were: to determine the in vitro antimicrobial activity of an ethanol extract of propolis; to define the MIC50 and MIC90 (Minimal Inhibitory Concentration - MIC) against 210 strains of S. aureus; to characterize a crude sample of propolis and the respective ethanol extract as to the presence of predetermined chemical markers. The agar dilution method was used to define the MIC and the high performance liquid chromatography (HPLC) method was used to characterize the samples of propolis. MIC results ranged from 710 to 2,850 µg/mL. The MIC50 and MIC90 for the 210 strains as well as the individual analysis of American Type Culture Collection (ATCC) strains of Methicillin-susceptible Staphylococcus aureus (MSSA) and Methicillin-resistant Staphylococcus aureus (MRSA) were both 1,420 µg/mL. Based on the chromatographic analysis of the crude sample and ethanol extracted propolis, it was concluded that propolis was a mixture of the BRP (SP/MG) and BRP (PR) types. The results obtained confirm an antimicrobial activity in relation to the strains of the S. aureus tested.

Brazilian propolis: correlation between chemical composition and antimicrobial activity.

The chemical composition of ethanol extracts from samples of Brazilian propolis (EEPs) determined by HPLC and their activity against Trypanosoma cruzi, Staphylococcus aureus, Streptococcus pneumoniae, Klebisiella pneumoniae, Candida albicans, Sporothrix schenckii and Paracoccidioides brasiliensis were determined. Based on the predominant botanical origin in the region of samples' collection, the 10 extracts were separated into three groups: A (B. dracunculifolia + Auraucaria spp), B (B. dracunculifolia) and C (Araucaria spp). Analysis by the multiple regression of all the extracts together showed a positive correlation, higher concentrations leading to higher biological effect, of S. aureus with p-coumaric acid (PCUM) and 3-(4-hydroxy-3-(oxo-butenyl)-phenylacrylic acid (DHCA1) and of trypomastigotes of T. cruzi with 3,5-diprenyl-4-hydroxycinnamic acid derivative 4 (DHCA4) and 2,2-dimethyl-6-carboxyethenyl-2H-1-benzopyran (DCBEN). When the same approach was employed for each group, due to the small number of observations, the statistical test gave unreliable results. However, an overall analysis revealed for group A an association of S. aureus with caffeic acid (CAF) and dicaffeoylquinic acid 3 (CAFQ3), of S. pneumoniae with CAFQ3 and monocaffeoylquinic acid 2 (CAFQ2) and of T. cruzi also with CAFQ3. For group B, a higher activity against S. pneumoniae was associated DCBEN and for T. cruzi with CAF. For group C no association was observed between the anitmicrobial effect and any component of the extracts. The present study reinforces the relevance of PCUM and derivatives, especially prenylated ones and also of caffeolyquinic acids, on the biological activity of Brazilian propolis.

Evaluation of the analgesic and anti-inflammatory effects of a Brazilian green propolis.

Phamacological activities of a standard ethanol extract G1 from Brazilian green propolis, typified as BRP1, was evaluated in mouse models of pain and inflammation. Intraperitoneal injection ( I. P.) of G1 inhibited acetic acid-induced abdominal constrictions with an ID (50) = 0.75 +/- 0.05 mg/kg, and in the formalin test the ID (50) values were 0.85 +/- 0.07 mg/kg and 13.88 +/- 1.12 mg/kg, respectively, for the neurogenic and inflammatory phases. The extract was ineffective when assessed in the hot-plate assay. In serotonin-induced paw edema, G1 led to a maximal inhibition (MI) of 51.6 % after 120 min when administered I. P. and of 36 % after 15 min by the oral route ( O. R.). When the inflammatory agent was complete Freund's adjuvant, inhibition of paw edema was also observed after administration of the extract by both routes. In the capsaicin-induced ear edema the ID (50) values were 1.09 +/- 0.08 mg/kg ( I. P.) and 10.00 +/- 0.90 mg/kg ( O. R.). In the acute carrageenan-induced inflammatory reaction induced by carrageenan, G1 reduced the number of neutrophils in the peritoneal cavity with IC (50) values of 0.72 +/- 0.08 mg/kg and 4.17 +/- 0.50 mg/kg, by I. P. or O. R. administration, with a preferential migration of polymorphonuclear neutrophils. IN VITRO, G1 decreased nitric oxide production in LPS-stimulated RAW 264.7 cells (IC (50) = 41.60 microg/mL), and also the luciferase activity in TNF-alpha-stimulated HEK 293 cells transfected with NF-kappaB-luciferase reporter gene driven by the nuclear factor kappaB (NF-kappaB) (IC (50) = 200 microg/mL). This extract, which at low concentrations induces anti-inflammatory and analgesic effects in mouse models, presents a high content of flavonoids, known to inhibit inducible NOS (iNOS) activity. These data taken together led us to reinforce the hypothesis in the literature that the anti-inflammatory effect of propolis may be a due to inhibition of iNOS gene expression, through interference with NF-kappaB sites in the iNOS promoter.

Bioactive constituents of brazilian red propolis.

In a new propolis type, red Brazilian propolis, 14 compounds were identified (six of them new for propolis), among them simple phenolics, triterepenoids, isoflavonoids, prenylated benzophenones and a naphthoquinone epoxide (isolated for the first time from a natural source). Three of the major components demonstrated significant antimicrobial activity, and two (obtained as inseparable mixture) possessed radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH).