Bioactives of Melipona rufiventris Propolis: Exploring its Antimicrobial, Anti-Inflammatory, and Antioxidant Activities.

This study explores the potential of propolis, a resinous substance produced by bees, from Melipona rufiventris species. With its composition encompassing resin, wax, pollen, and soil, propolis holds historical significance in traditional medicine within tropical regions. This research is driven by the scarcity of information surrounding M. rufiventris propolis, prompting an investigation into its chemical constituents, in vivo toxicity, and antimicrobial, antioxidant, and anti-inflammatory properties. This exploration could potentially uncover novel applications for this natural product, bolstering both meliponiculture practices and the preservation of native bee populations. The propolis was sampled in Cabo Verde-MG and underwent ethanolic extraction to yield an extract (EEP) for analysis. Chemical assessments (Folin-Ciocalteau, and UHPLC-HRMS) revealed the presence of polyphenols, including flavonoids. The EEP demonstrated higher antimicrobial activity against Gram-positive bacteria and exhibited efficacy against multiresistant strains isolated from complex wounds. Synergistic interactions with commercial antibiotics were also observed. Furthermore, anti-inflammatory evaluations showcased the EEP's potential in reducing NF-kB activation and TNF-α release at non-toxic concentrations. Despite these promising biological activities, the EEP exhibited no antiproliferative effects and demonstrated safety in both the MTS assay and the G. mellonella model. Collectively, these findings highlight the M. rufiventris propolis extract as a valuable reservoir of bioactive compounds with multifaceted potential.

Evaluation of a Standardized Extract Obtained from Cashew Apple (Anacardium occidentale L.) Bagasse in DSS-Induced Mouse Colitis.

Inflammatory bowel diseases (IBD) include Crohn's disease and ulcerative colitis. Several studies relate eating habits to different aspects of IBD, such as progression and worsening of the clinical condition. Therefore, many natural products (NPs) such as polyphenols and carotenoids have been identified as promising agents in supporting IBD. An interesting source for obtaining bioactive NPs is the by-products of the food industry. The present study evaluated the potential beneficial effect of a standardized extract (CAE) obtained from cashew apple bagasse in the dextran sulfate sodium (DSS)-induced ulcerative colitis model in mice. This was the first time that CAE had been evaluated in this experimental model. Chemical evaluation of CAE identified carotenoids (96.28 ± 0.15 mg/100 g), phenolic compounds (37.49 ± 0.64 mg/100 g), and a mixture of anacardic acids (C15:3 = 94.2 ± 0.6 mg/100 g; C15:2 = 108.4 ± 0.1 mg/100 g; C15:1 = 214.8 ± 0.2 mg/100 g). Administration of CAE (500 mg/kg, 4 days, p.o.) after DSS challenge was more effective in delaying disease progression compared with prior treatment (500 mg/kg, 30 days, p.o.), according to the disease activity index. However, no treatment strategy with CAE was able to prevent or inhibit disease progression, since all parameters evaluated (macroscopic, biochemical, and histopathological) in CAE-treated animals were similar to those observed in DSS-challenged animals. Despite the high dose (500 mg/kg), the standardized extract (CAE) did not result in an effective concentration of carotenoids. Furthermore, as some anacardic acids have been reported as histone acetyltransferases inhibitors, there could be a possible antagonistic relationship between carotenoids and anacardic acids. Complementary research will be necessary to test the hypothesis of antagonism. Thus, an optimized extract, with an even higher concentration of carotenoids, obtained from cashew apple bagasse, can be developed as a possible adjuvant food supplement for inflammatory bowel diseases.

Itraconazole and neutrophil interactions in the immune-inflammatory response of paracoccidioidomycosis using a murine air pouch infection model.

Paracoccidioidomycosis (PCM) caused by Paracoccidioides brasiliensis (Pb), is a severe mycosis, prevalent in tropical countries. The presence of polymorphonuclear neutrophils (PMN) in lesions is conspicuous, indicating their central role in innate immunity through the direct killing of Pb and the production of cytokines that activate acquired immunity in the presence of itraconazole (Itra). The toxicity and direct antifungal activity of Itra on Pb in splenocyte co-cultures were evaluated in vitro. Itra showed no toxic effect but marked antifungal activity against Pb. Purified PMN were obtained by the subcutaneous (SC) injection of Pb into mice. Results showed the effect of Itra on the size of the air pouch produced, the cellular population that migrated to the infection site, protein, and mitochondrial metabolism patterns, production of ROS an NO, and the number of cytokines synthesized. Lower doses (3 and 10 mg/kg) of Itra did not affect the cellular profile but led to a lower influx of viable more active PMN, and increased production of ROS and proteins. At a dose of 50 mg/kg the PMN profile remained unchanged along with all other parameters analyzed remained unaltered. Decreases in most cytokine levels were inversely proportional to the Itra concentration. Lower Itra concentrations may elicit activation of the immune response because the combined effects of therapy and immune response are needed, while the higher dose does not require it. Itra also promotes the activation of the cytokines which elicit PMN activation and consequently the resolution of Pb18 infection in the air pouch.

Brazilian organic propolis for prevention and treatment of radiation-related oral acute toxicities in head and neck cancer patients: A double-blind randomized clinical trial.

Background: Oral mucositis (OM) is one of the most important acute toxicities from radiotherapy (RT) in head and neck cancer patients and can impair oncologic treatment. Dysphagia, dysgeusia, pain, and oral candidiasis are other common toxicities. Brazilian Organic Propolis (BOP) is a recently described propolis variant and BOP types 4 and 6 have shown important antioxidant, anti-inflammatory, and antifungal properties. Purpose: To investigate the use of BOP as a preventive and/or complementary therapeutic option for radiotherapy-induced oral mucositis, dysphagia, dysgeusia, pain, and oral candidiasis. Additionally, proinflammatory cytokines were assessed to investigate their anti-inflammatory role. Methods: Sixty patients were included in this randomized, double-blind, controlled clinical trial. Patients were randomized to receive either aqueous suspension of a BOP or placebo throughout RT. Also, all patients underwent low-level laser therapy as routine oral care. OM, dysphagia, and dysgeusia were assessed weekly according to WHO and NCI scales. Pain-related to OM was assessed according to a Visual Analog Scale and the presence or absence of oral candidiasis was checked by intraoral examination. Protein levels of TNF-α and IL-1ß from oral mucosa were assessed by ELISA. Results: Patients in the propolis group had a lower mean score of OM, dysphagia, dysgeusia, and most patients reported moderate pain. Fewer patients developed oral candidiasis in the propolis group, and the number of episodes was lower among patients that used BOP (p < 0.05). In addition, the BOP group presented significantly lower levels of IL-1ß since the beginning of treatment when compared with placebo patients (p < 0.05) and a lower level of TNF-α at the end of treatment (p < 0.001). Conclusion: Topic use of BOP reduced TNF-α and IL-1ß levels, oral candidiasis episodes, and seems to be a useful complementary option for the prevention and treatment of the main acute oral toxicities of RT. Clinical Trial Registration: http://www.ensaiosclinicos.gov.br/rg/RBR-9f8c78/, identifier RBR-9f8c78.

Phenolic Profile and the Antioxidant, Anti-Inflammatory, and Antimicrobial Properties of Açaí (Euterpe oleracea) Meal: A Prospective Study.

The mechanical extraction of oils from Brazilian açaí (Euterpe oleracea Mart) produces significant amounts of a byproduct known as "meal", which is frequently discarded in the environment as waste material. Nevertheless, plant byproducts, especially those from oil extraction, may contain residual polyphenols in their composition and be a rich source of natural bioactive compounds. In this study, the phenolic composition and in vitro biological properties of a hydroethanolic açaí meal extract were elucidated. The major compounds tentatively identified in the extract by high-resolution mass spectrometry were anthocyanins, flavones, and flavonoids. Furthermore, rhamnocitrin is reported in an açaí byproduct for the first time. The extract showed reducing power and was effective in scavenging the ABTS radical cation (820.0 µmol Trolox equivalent∙g-1) and peroxyl radical (975.7 µmol Trolox equivalent∙g-1). NF-κB activation was inhibited at 10 or 100 µg∙mL-1 and TNF-α levels were reduced at 100 µg∙mL-1. However, the antibacterial effects against ESKAPE pathogens was not promising due to the high concentration needed (1250 or 2500 µg∙mL-1). These findings can be related to the diverse polyphenol-rich extract composition. To conclude, the polyphenol-rich extract obtained from açaí meal showed relevant biological activities that may have great applicability in the food and nutraceutical industries.

Biological properties of Schinus terebinthifolia Raddi essential oil

Abstract Schinus terebinthifolia Raddi green fruits essential oil (EO) was evaluated regarding its phytochemical profile, antimicrobial and cytotoxic activities, and toxicity. Gas chromatography with mass spectrometry was applied to identify its constituents, thereafter the minimum inhibitory concentration, minimum bactericidal and fungicidal concentrations, and its antibiofilm activity were evaluated. The EO cytotoxicity was assessed in tumor and non-tumor human cells, and in vivo toxicity was evaluated in a Galleria mellonella model. The major constituents of S. terebinthifolia EO were alpha-phellandrene and beta-phellandrene. The EO had a weak activity against all strains of Candida albicans (MIC 1000µg/mL) and had no activity against non-albicans strains, bacteria, and C. albicans biofilm. Cytostatic activity against all tumor cell lines was shown. Additionally, cell viability remained at EO concentrations up to 62.5 µg/mL. At 16 mg/mL, 50% hemolysis was observed, and it had low toxicity in vivo. Overall, the S. terebinthifolia EO was characterized by low antimicrobial and antibiofilm activities, with no evidence of toxicity to human tumor and non-tumor cells

Phytochemical composition, antifungal activity, in vitro and in vivo toxicity of Syzygium cumini (L) Skeels leaves extract / Composición fitoquímica, actividad antifúngica, toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (L) Skeels

This study determined phytochemical composition, antifungal activity and toxicity in vitro and in vivo of Syzygium cumini leaves extract (Sc). Thus, was characterized by gas chromatography coupled to mass spectrometry and submitted to determination of Minimum Inhibitory (MIC) and Fungicidal concentrations (MFC) on reference and clinical strains of Candida spp. and by growth kinetics assays. Toxicity was verified using in vitro assays of hemolysis, osmotic fragility, oxidant and antioxidant activity in human erythrocytes and by in vivo acute systemic toxicity in Galleria mellonella larvae. Fourteen different compounds were identified in Sc, which showed antifungal activity (MIC between 31.25-125µg/mL) with fungistatic effect on Candida. At antifungal concentrations, it demonstrated low cytotoxicity, antioxidant activity and neglible in vivotoxicity. Thus, Sc demonstrated a promising antifungal potential, with low toxicity, indicating that this extract can be a safe and effective alternative antifungal agent.

Este estudio determinó la composición fitoquímica, la actividad antifúngica y la toxicidad in vitro e in vivo del extracto de hojas de Syzygium cumini (Sc). Así, se caracterizó mediante cromatografía de gases acoplada a espectrometría de masas y se sometió a determinación de Concentraciones Mínimas Inhibitorias (CMI) y Fungicidas (MFC) sobre cepas de referencia y clínicas de Candida spp. y mediante ensayos de cinética de crecimiento. La toxicidad se verificó mediante ensayos in vitro de hemólisis, fragilidad osmótica, actividad oxidante y antioxidante en eritrocitos humanos y por toxicidad sistémica aguda in vivo en larvas de Galleria mellonella. Se identificaron catorce compuestos diferentes en Sc, que mostraron actividad antifúngica (CMI entre 31.25-125 µg/mL) con efecto fungistático sobre Candida. En concentraciones antifúngicas, demostró baja citotoxicidad, actividad antioxidante y toxicidad in vivo insignificante. Por lo tanto, Sc demostró un potencial antifúngico prometedor, con baja toxicidad, lo que indica que este extracto puede ser un agente antifúngico alternativo seguro y eficaz.

Isolation of diterpenes from Araucaria sp Brazilian brown propolis and development of a validated high-performance liquid chromatography method for its analysis.

Propolis comprises a complex resinous product composed of plant's parts or exudates, pollen, bee wax, and enzymes. Brazilian brown propolis from Araucaria sp displays several biological activities. Considering the lack of validated analytical methods for its analysis, we are reporting the development of a validated high-performance liquid chromatography with photodiode array detector method to analyze Araucaria brown propolis. The crude propolis were extracted and chromatographed, furnishing six main diterpenes. The isolated standards were used to draw the analytical curves, allowing the studies of selectivity, precision, accuracy, recovery, robustness, the determination of limits of detection and limits of quantification. The mobile phase consisted of 0.1% acetic acid in water and acetonitrile, using an octadecylsilane column, 1 mL/min flow rate and detection at 200 or 241 nm. Relative standard deviation values obtained for intra-day and inter-day precision were lower than 4% for all diterpenes. From the five parameters for robustness, wavelength detection and flow rate were the critical ones. Limits of detection and quantification ranged from 0.808 to 10.359 µg/mL and from 2.448 to 31.392 µg/mL, respectively. The recoveries were between 105.03 and 108.13%, with relative standard deviation values around 5.0%. The developed method is precise, sensitive, and reliable for analyzing Araucaria brown propolis.

Inajá oil processing by-product: A novel source of bioactive catechins and procyanidins from a Brazilian native fruit.

Agro-industrial activities generate large amounts of solid residues, which are generally discarded or used as animal feed. Interestingly, some of these by-products could serve as natural sources of bioactive compounds with great potential for industrial exploitation. This study aimed to optimize the extraction of phenolic antioxidants from the pulp residue (oil processing by-product) of inajá (Maximiliana maripa, a native species found in the Brazilian Amazon). The antioxidant properties of the optimized extract and its phenolic profile by high-resolution mass spectrometry (LC-ESI-QTOF-MS) were further determined. Central composite rotatable design and statistical analysis demonstrated that the temperature of 70 °C and 50% (v/v) ethanol concentration improved the extraction of phenolic compounds with antioxidant properties. The optimized extract also showed scavenging activity against the ABTS radical cation and reactive oxygen species (ROS; peroxyl and superoxide radical, and hypochlorous acid). Moreover, the optimized extract was able to reduce NF-κB activation and TNF-α release, which are modulated by ROS. Flavan-3-ols were the major phenolics present in the optimized extract. Collectively, our findings support the use of inajá cake as a new source of bioactive catechins and procyanidins. This innovative approach adds value to this agro-industrial by-product in the functional food, nutraceutical, pharmaceutical, and/or cosmetic industries and complies with the circular economy agenda.

Brazilian Red Propolis shows antifungal and immunomodulatory activities against Paracoccidioides brasiliensis.

ETHNOPHARMACOLOGICAL RELEVANCE: Paracoccidioidomycosis (PCM) is a systemic mycosis with high prevalence in South America and especially in Brazil with severe clinical consequences that need broadened therapeutic options. Propolis is a natural resin from bees used in folk medicine for centuries with the first report in the ancient history of Egypt by Eberly papyrus, in Middle-Ages used to wash the newborn's umbilical cord and World War II as antiseptic or antibiotics. Nowadays it is a natural product worldwide consumed as food and traditionally used for oral and systemic diseases as an anti-inflammatory, antimicrobial, antifungal, and other diseases. Brazilian red propolis (BRP) is a new type of propolis with a distinguished chemical profile and biological activities from propolis (green) with pharmacological properties such as antimicrobial, anti-inflammatory, antioxidant, and others. AIM OF STUDY: Thus, the main purpose of this study was to investigate the direct in vitro and ex vivo effect of BRP on Paracoccidioides brasiliensis. MATERIAL AND METHODS: Antifungal activity of different concentrations of BRP on a virulent P. brasiliensis isolate (Pb18) was evaluated using the microdilution technique. Also, mice splenic cells co-cultured with Pb18 were treated with BRP at different times and concentrations (only Pb18 = negative control). Mice were inoculated with Pb18 and treated with different concentrations of BRP (50-500 mg/mL) in a subcutaneous air pouch. In this later experimental model, macroscopic characteristics of the air pouch were evaluated, and cellular exudate was collected and analyzed for cellular composition, mitochondrial activity, total protein reactive oxygen specimens (ROS), and nitric oxide production, as well as the number of viable fungal cells. RESULTS: The in vitro experiments showed remarkable direct antifungal activity of BRP, mainly with the highest concentration employed (500 mg/mL), reducing the number of viable cells to 10% of the original inoculum after 72 h incubation. The splenocytes co-cultivation assays showed that BRP had no cytotoxic effect on these cells, on the contrary, exerted a stimulatory effect. This stimulation was also observed on the PMNs at the air pouch, as verified by production of ROS and total proteins and mitochondrial activity. This activation resulted in enhanced fungicidal activity, mainly with the 500 mg/mL concentration of BRP. An anti-inflammatory effect was also detected, as verified by the smaller volume of the BRP-treated air pouch as well as by an earlier shift from neutrophils to mononuclear cells present in the infection site. CONCLUSION: Our results strongly suggest, for the first time in the literature, that Brazilian Red propolis has four protective mechanisms in experimental paracoccidioidomycosis: activating neutrophils, exerting a direct antifungal effect, preventing fungal dissemination, and controlling excessive inflammation process.

Cashew apple byproduct: Gastroprotective effects of standardized extract.

ETHNOPHARMACOLOGICAL RELEVANCE: The incidence of gastric mucosa lesions in the adult population has increased mainly due to the continued use of nonsteroidal anti-inflammatory drugs (NSAIDs). The cashew (Anacardium occidentale L.) is a tropical tree, cultivated in several countries, whose barks, leaves and pseudofruit (cashew apple) are popularly used in traditional medicine for the treatment of many diseases, including gastric ulcer. AIM: Our study evaluated the potential gastroprotective effect of the carotenoid and anacardic acids-enriched aqueous extract (CAE), prepared from cashew apple pomace, in the dose-repeated acetylsalicylic acid (ASA)-induced gastric lesions model in rats. MATERIAL AND METHODS: After randomly distribution into five group (G1 - G5, n = 8 animals/group), male Wistar rats were daily treated with ASA solution (200 mg/kg, 5 ml/kg, G2 - G5) or potable water (Satellite group, G1) during 14 days. From 8th to 14th experimental day, rats in G3 - G5 groups were orally treated with CAE (50, 100 and 500 mg/kg, 5 ml/kg, respectively). Body weight was measured on 0, 7th and 14th day. On the 14th experimental day, all surviving animals were euthanized for macroscopic evaluation of the inner organs and stomach removal. After weighting, each stomach was properly prepared for biochemical analysis [myeloperoxidase activity (MPO), reduced glutathione analysis (GSH), IL-1ß, CXCL2/MIP-2, TNF-α and IL-10 levels]. RESULTS: At the most efficient dose (100 mg/kg, p.o.), CAE-treated animals showed a slight improvement in the macroscopic aspect of gastric mucosa associated with significant (p < 0.05) reduced levels of IL-1ß, CXCL2/MIP-2, and MPO activity besides increased levels of GSH (partially), and IL-10 in stomach tissues. CONCLUSIONS: The present study demonstrated that the carotenoid and anacardic acids-enriched extract obtained from cashew apple pomace is a promising raw material for the development of herbal medicine and/or functional food supplements for the adjuvant treatment of NSAIDs-induced gastric ulcers.

Assessment of a novel bleaching agent formula containing 35% hydrogen peroxide and titanium tetrafluoride: an in vitro study

Abstract: This study developed experimental gels containing titanium tetrafluoride (TiF4) combined with commercial 35% hydrogen peroxide (HP), and evaluated bleaching efficacy and pH of the gels, and mineral content and morphology of enamel submitted to these treatments. In phase-1, different stock gels mixed with TiF4 were combined with HP. In phase-2, the selected gels were tested on enamel/dentin specimens (n=8): HP; HP and Natrosol+TiF4 (HPnT); HP and Natrosol+Chemygel+TiF4 (HPncT); HP and Aristoflex+TiF4 (HPaT). Bleaching was performed in four sessions (3x15min-application/session). Color (CIEL*a*b*) and whiteness index (WID) were measured after each session, whereas whiteness index differences (ΔWID), color alteration (CIELab-ΔE, CIEDE2000-ΔE00), enamel morphology and pH, at end of bleaching therapy. The change in Knoop microhardness (ΔKHN) was compared before and after bleaching. Data were analyzed by two-way repeated measures ANOVA and Bonferroni (CIEL*, a*, b*), one-way ANOVA and Tukey (ΔWID, ΔE, ΔE00), and LSD (ΔKHN) tests (α=5%). SEM and pH measurements were submitted to descriptive analysis. No differences were observed in lightness (L*) or WID among the groups (p > 0.05), but HP exhibited lower b* values (p<0.05), higher ΔWID than HPnT, and the highest ΔE among the groups (p < 0.05). No differences in ΔE00 were observed between HP and HPncT (p > 0.05), and HPncT showed higher ΔKHN than HP (p < 0.05). HP presented pH values closer to neutral (6.9), whereas experimental agents showed acidic pH values (2.3-3.9). No morphological changes were observed in HP or HPncT groups. HPncT was able to bleach the enamel and maintain enamel microhardness and surface integrity, even at low pH.

Simulated gastrointestinal digestion of Brazilian açaí seeds affects the content of flavan-3-ol derivatives, and their antioxidant and anti-inflammatory activities.

Açaí seeds (Euterpe oleracea Mart.) are the major residue generated during industrial extraction of açaí fruit pulp - a popular and typical Amazon fruit rich in bioactive compounds and nutrients. In this study, we investigated the bioaccessibility of an açaí seed extract using an in vitro simulated gastrointestinal digestion model. Catechin, epicatechin and procyanidins B1 and B2 were identified and quantified in the açaí seed extract and monitored by HPLC-DAD through the digestion phases. Bioaccessibility of these flavan-3-ols and deactivation of reactive oxygen species decreased after the intestinal phase, except for peroxyl radical (ROO●). RAW 264.7 macrophages treated either with the digested or undigested açaí seed extract showed reduced NF-κB activation and TNF-α levels, even following gastrointestinal digestion. Thus, the ROO● scavenging capacity and anti-inflammatory activity of the extract were found to be still remarkable after digestion, suggesting that açaí seeds could be explored as a source of bioactive compounds for functional foods, cosmetic or pharmaceutical purposes.

Phenolic profile and potential beneficial effects of underutilized Brazilian native fruits on scavenging of ROS and RNS and anti-inflammatory and antimicrobial properties.

Brazilian native fruits (BNF) have aroused interest of researchers and consumers for their great human health benefits. In this study, five BNF (Byrsonima lancifolia, Campomanesia phaea, Jacaratia spinosa, Solanum alternatopinnatum and Acnistus arborescens) were tested for their polyphenolic compounds by LC-ESI-MS/MS, reactive species deactivation (ROO˙, O2˙-, HOCl and NO˙), anti-inflammatory properties in vivo, and in vitro antimicrobial activity - with determination of putative mechanism(s) of action. Eighty-one polyphenols were identified, which exhibited a significant capacity to deactivate both ROS and RNS. C. phaea extract had the highest capacity to scavenge ROO˙ (68.94 µmol TE per g), O2˙- (IC50: 575.36 µg mL-1) and NO˙ (IC50: 16.96 µg mL-1), which may be attributed to the presence of ellagitanins. B. lancifolia decreased neutrophil influx into the peritoneal cavity of mice by 50% as compared to carrageenan and reduced Candida albicans biofilm viability by 3 log10 possibly due to complexation with cell membrane ergosterol. In summary, the BNF presented herein are good sources of bioactive compounds with positive effects on deactivation of biological reactive species, as well as with anti-inflammatory and antimicrobial activities, which can be altogether highly beneficial to human health.

Effect of Cinnamomum verum leaf essential oil on virulence factors of Candida species and determination of the in-vivo toxicity with Galleria mellonella model.

BACKGROUND: Essential oils (EO) extracted from Cinnamomum verum has been used as an antimicrobial agents for centuries. The effects of C. verum leaf oil against virulence of microorganisms is not well studied yet. OBJECTIVES: This study evaluates the effect of C. verum leaf oil against three virulence factors of Candida albicans, C. tropicalis and C. dubliniensis and its in-vivo toxicity. METHODS: Chemical composition of EO was determined using gas chromatography-mass spectrometry (GC-MS). Minimum inhibitory concentration (MIC) was determined using clinical and laboratory standards institute (CLSI) M27-A3 broth microdilution. Effect of EO on initial adhesion was quantified using XTT assay after allowing Candida cells to adhere to the polystyrene surface for 2 h. Biofilm formation of Candida in the presence of EO was quantified using XTT viability assay. Efficacy on reduction of germ tube formation was evaluated using standard protocol. Visualisation of biofilm formation and progression under the EO treatment were done using scanning electron microscope (SEM) and Time lapses microscope respectively. In-vivo toxicity of EO was determined using Galleria mellonella larvae. Chlorhexidine digluconate: positive control. RESULTS: Eugenol was the main compound of EO. MIC was 1.0 mg/mL. 50% reduction in initial adhesion was achieved by C. albicans, C. tropicalis and C. dubliniensis with 1.0, > 2.0 and 0.34 mg/mL respectively. 0.5 and 1.0 mg/mL significantly inhibit the germ tube formation. MBIC50 for forming biofilms were ≤ 0.35 mg/mL. 1.0 mg/mL prevent biofilm progression of Candida. SEM images exhibited cell wall damages, cellular shrinkages and decreased hyphal formation. No lethal effect was noted with in-vivo experiment model at any concentration tested. CONCLUSION: C. verum leaf oil acts against virulence factors of Candida and does not show any toxicity.

Anti-inflammatory and antioxidant potential, in vivo toxicity, and polyphenolic composition of Eugenia selloi B.D.Jacks. (pitangatuba), a Brazilian native fruit.

Brazilian native fruits are a rich source of polyphenolic compounds that can act as anti-inflammatory and antioxidant agents. Here, we determined the polyphenolic composition, anti-inflammatory mechanism of action, antioxidant activity and systemic toxicity in Galleria mellonella larvae of Eugenia selloi B.D.Jacks. (synonym Eugenia neonitida Sobral) extract (Ese) and its polyphenol-rich fraction (F3) obtained through bioassay-guided fractionation. Phenolic compounds present in Ese and F3 were identified by LC-ESI-QTOF-MS. The anti-inflammatory activity of Ese and F3 was tested in vitro and in vivo through NF-κB activation, cytokine release and neutrophil migration assays. The samples were tested for their effects against reactive species (ROO•, O2•-, HOCl and NO•) and for their toxicity in Galleria mellonella larvae model. The presence of hydroxybenzoic acid, ellagitannins and flavonoids was identified. Ese and F3 reduced NF-κB activation, cytokine release and neutrophil migration, with F3 being three-fold more potent. Overall, F3 exhibited strong antioxidant effects against biologically relevant radicals, and neither Ese nor F3 were toxic to G. mellonella larvae. In conclusion, Ese and F3 revealed the presence of phenolic compounds that decreased the inflammatory parameters evaluated and inhibited reactive oxygen/nitrogen species. E. selloi is a novel source of bioactive compounds that may provide benefits for human health.

Anti-inflammatory and anti-Candida Effects of Brazilian Organic Propolis, a Promising Source of Bioactive Molecules and Functional Food.

Brazilian organic propolis (BOP) is an unexplored Brazilian propolis that is produced organically and certified according to international legislation. Our results showed that BOP has strong anti-inflammatory effects and acts by reducing nuclear factor κB activation, tumor necrosis factor α release, and neutrophil migration. In addition, BOP6 exhibited antifungal activity on planktonic and biofilm cultures of Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei, and Candida parapsisolis and reduced in vitro yeast cell adhesion to human keratinocytes at sub-inhibitory concentrations. BOP demonstrated significantly low toxicity in Galleria melonella larvae at antifungal doses. Lastly, a chemical analysis revealed the presence of caffeoyltartaric acid, 3,4-dicaffeoylquinic acid, quercetin, and gibberellins A7, A9, and A20, which may be responsible for the biological properties observed. Thus, our data indicate that BOP is a promising source of anti-inflammatory and antifungal molecules that may be used as a functional food.

Optimizing the potential bioactivity of isoflavones from soybeans via ultrasound pretreatment: Antioxidant potential and NF-κB activation.

Soybean consumption has been associated with health benefits. However, the effect of ultrasound (US) soybean pretreatment in terms of potential health benefits has not been investigated so far. Accordingly, the total phenolic content (TPC) and the total aglycone content (TAC) were optimized using the Box-Behnken design. Contrasting samples regarding isoflavones aglycones and TPCs were screened for their antioxidant and anti-inflammatory potentials using RAW 264.7 macrophages. US pretreated soybeans (55°C, 15 min, and 24 W/cm2 ) showed greater TPC and TAC compared to the control and this translated to higher antiradical activity and reduction of nuclear factor kappa B (NF-κB) activation. The concentration of genistein in treated soybeans increased by 95%. Furthermore, US pretreated soybeans rendered phenolic extracts that reduced the NF-κB activation by 86%. Therefore, this contribution demonstrates the beneficial effects of US pretreatment of soybeans, which provides a better feedstock for the functional food industry. PRACTICAL APPLICATIONS: Soybeans can be consumed as such or used as a feedstock to produce soy yogurt, fermented soymilk, tofu, and protein concentrate, among others. The greatest bioavailability of isoflavones compared to other flavonoids has recently been highlighted, and this has been explained by the relatively moderate lipophilicity of isoflavones as aglycones. The present contribution supports the use of US pretreatment of soybeans to obtain a feedstock with improved contents of isoflavones as aglycones. We have confirmed that phenolic extracts obtained from the US pretreated samples showed higher bioactivity as radical scavengers and by reducing the activation of nuclear factor kappa B (NF-κB) in a cell model, which is mediated by oxidative species. The clinical importance of NF-κB activation is derived mainly from its role in inflammatory responses. Therefore, our investigation may have a practical application in the procurement of soybean products and/or ingredients with improved functional properties related to their health benefits.

Antifungal, antibiofilm, and antiproliferative activities of Guapira graciliflora Mart.

The aim of this study was to evaluate in vitro the antifungal, antibiofilm and antiproliferative activities of the extract from the leaves of Guapira graciliflora Mart. The phytochemical characterization of the extract was performed using electrospray ionization mass spectrometry (ESI-MS). The antimicrobial activity of the extract and its fractions was evaluated using the broth microdilution method against species of Candida. The inhibition of C. albicans biofilm was evaluated based on the number of colony-forming units (CFU) and metabolic activity (MTT). The antiproliferative activity of the extract and its fraction was evaluated in the presence of human tumor and non-tumor cells, and the cytotoxicity of the extract was determined on the RAW 264.7 macrophage line - both using the sulforhodamine B method. The phytochemical characterization indicated the presence of the flavonoids rutin and kaempferol. The extract and the methanol fraction exhibited moderate antifungal activity against C. albicans, C. krusei, and C. glabrata, and strong activity against C. dubliniensis. In the biofilms at 24 and 48 hours, the concentration of 12500 µg/mL of the extract was the most effective at reducing the number of CFU s/mL (44.4% and 42.9%, respectively) and the metabolic activity of C. albicans cells (34.6% and 52%, respectively). The extract and its fractions had no antiproliferative effect on the tumor lines tested, with mean activity (log GI50) equal to or greater than 1.71 µg/mL. Macrophage cell viability remained higher than 80% for concentrations of the extract of up to 62.5 µg/mL. G. graciliflora has flavonoids in its chemical composition and demonstrates potential antifungal and antibiofilm activity, with no evidence of a significant change in the viability of human tumor and non-tumor cell lines.

Bioprospection of Eugenia brasiliensis, a Brazilian native fruit, as a source of anti-inflammatory and antibiofilm compounds.

The anti-inflammatory and antibiofilm activities as well as toxicity and chemical profile of Eugenia brasiliensis pulp extract (EBE), were evaluated. EBE chemical profile and phenolic content were determined by LC-MS/MS. EBE was tested for its in vitro and in vivo anti-inflammatory activity, including TNF-α release, NF-кB activation, neutrophil migration and paw edema. The MIC/MBC and antibiofilm activities were tested against methicillin sensitive and resistant Staphylococcus aureus, Escherichia coli, Pseudomona aeruginosa, Streptococcus mutans, and Lactobacillus acidophilus. EBE acute toxicity was evaluated in Galleria mellonella and RAW 264.7 macrophage. EBE total phenolic content was 389.88 ±â€¯3.48 mg GAE/g with identified polyphenols. EBE decreased TNF-α release in vivo and in vitro, NF-кB activation, neutrophil influx into peritoneal cavity, and it showed maximal inhibition of paw edema after 2 h. MIC of EBE ranged from 62.5-500 µg/mL while MBC values were >500 µg/mL, with a decrease in L. acidophilus biofilm formation. EBE showed negligible toxicity in larvae and macrophage cells. Our findings open new perspectives concerning EBE application as source of anti-inflammatory and antibiofilm molecules as a functional food, pharmaceutical lead or agribusiness commodity.