Polymeric Nanoparticles for the Treatment of Prostate Cancer- Technological Prospecting and Critical Analysis.

BACKGROUND: Polymeric nanoparticles have a wide diversity, and due to their toxicity and biodegradability, they have been widely used in the health area. Its use allows stability of some compounds, targeted delivery, and increased half-life, in this context, making some treatment proposals more effective. Prostate cancer, in turn, is among the types of cancer with the highest mortality, and the lack of effective treatment causes several strategies to meet this need. OBJECTIVE: The objective of this work was to verify patents that use polymeric nanoparticles for the treatment of prostate cancer. METHODS: For that, specific keywords to direct the search were applied in Patent Scope. After obtaining the patents, one was selected for the development of critical analysis in relation to its chemistry and biology. RESULTS: A total of five patents were found and, of these, an invention that used PCL-PLGA-PEGCOOH polymeric nanoparticles with two natural compounds, resveratrol and celastrol, providing an alternative method to traditional monotherapies. CONCLUSION: The prospective analysis serves to direct us in relation to the technologies currently used in certain fields. Based on several cases of cancer and specifically the countless cases of prostate cancer, five recent patents were found using polymeric nanoparticles. However, only one brought a different aspect of all the treatments used lately. It brought an invention containing two natural compounds being carried with polymeric nanoparticles with promising results.

Chitosan Oligomer and Zinc Oxide Nanoparticles for Treating Wastewaters: US20190134086 Patent Evaluation.

With the utilisation of algae, wastewater reuse is becoming a viable option for the energy industry, especially green energy. The growth of these algae in these wastewaters provides an alternative source for bioenergetics, however, the growth of other microorganisms can directly affect the production of bioenergy, requiring the removal and reduction of contaminants in these waters, in addition to being a source of contamination for workers. Therefore, the use of nanoparticles in bioremediation has been an alternative to mitigate the contamination of these wastewaters that have microorganisms capable of reducing the algae growth capacity. The objective of this work was to verify in the United States Patent and Trademarker office database (USPTO) patents that used chitosan nanoparticles as a form of wastewater treatment and to carry out the analysis of patent US20190134086, which addresses the use of zinc oxide nanoparticles associated with chitosan that was developed and used to evaluate their antibacterial activity against resistant microorganisms and biofilm producers present in wastewater. Escherichia coli, Enterococcus faecium, and/or Pseudomonas aeruginosa are the microorganisms involved in the evaluated invention, bacteria present in the gastrointestinal tract, of clinical and environmental importance. The synthesized nanoparticles are arranged as a pharmaceutically acceptable and toxic vehicle against resistant bacteria, thus being described as nanoremediators. Given the analyzed patent, it was possible to verify the importance of alternatives to reduce the impact that pollution, in general, has on the environment, in addition to the proposed technology serving to maintain the survival and development capacity of the algae that will be able to produce green energy, the nanoparticles with antibacterial potential can help indirectly reduce these pathogenic strains with resistance to several antibiotics in the environment.

Peptide vaccines designed with the aid of immunoinformatic against Caseous Lymphadenitis promotes humoral and cellular response induction in mice.

Caseous Lymphadenitis (CLA) is a chronic disease that affects also small ruminants. CLA is caused by Corynebacterium pseudotuberculosis and is responsible for high economic losses due to the formation of superficial and visceral granulomas, the latter is considered as asymptomatic CLA causing high levels of dissemination. Several vaccination strategies, in which the use of synthetic peptides stands out. Thus, this work aimed to evaluate the protective potential of peptide vaccines designed to determine the immunodominant epitopes of CP40 against CLA in mice. The animals were divided into eight groups separated in controls (G1-PBS, G2-Saponin and G9-rCP40) and experimental (G3-pep1, G4- pep2, G5-pep3, G6-pep4, G7-pep5 and G8-pep6), these were vaccinated on days 0 and 15 by a subcutaneous route. 60 days after the first immunization, all animals were challenged with C. pseudotuberculosis. On days 0, 15, 60, and 120 after the first immunization, blood samples were taken to measure immunoglobulins. On the same day of the challenge, the splenocytes were isolated and assayed for the production of IL-2, IL-4, IL-6, IFN-γ, TNF-α, IL-17, and IL-10. After vaccinations, the animals were challenged and all of them were affected by the disease which led to their death. The G6 and G8 groups provided 10% protection and the G7 provided 20%. The G3 and G4 groups provided 30% and 40% protection respectively. The peptides showed the production of Total IgG antibodies and cytokines (IL-2, IL-4, IL-6, IFN-γ, and TNF-α), indicating a possible activation of the Th1 type response. However, groups G3, G5, G6, and G8 showed production of IL-17. None of the study groups showed IL-10 production. The immunogenicity of the peptides was not enough to protect these animals and it is believed that the use of adjuvants based on PAMPs may improve the immune response offered by these peptides.

In Silico Screening of Putative Corynebacterium pseudotuberculosis Antigens and Serological Diagnosis for Caseous Lymphadenitis in Sheep by Enzyme-Linked Immunosorbent Assay.

Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis (CLA), a disease leading to severe damage in sheep and goats farming due to the lack of serological diagnosis, treatment, and effective prophylaxis. In this context, several strategies in an attempt to discover new antigens to compose diagnosis assays or vaccines are fundamental. Therefore, this study aimed to use bioinformatics software to evaluate the critical chemical characteristics of unknown proteins of C. pseudotuberculosis by selecting them for heterologous expression in Escherichia coli. For this purpose, six protein sequences of ascorbate transporter subunit, UPF protein, MMPL family transporter, Ribonuclease, Iron ABC transporter domain-containing permease, and fimbrial subunit were obtained. In silico analyses were performed using amino acid sequences to access immunodominant epitopes and their antigenic and allergenic potential and physicochemical characterization. The expressed proteins were used as an antigen for serological diagnosis by ELISA. All proteins showed distinct immunodominant epitopes and potential antigenic characteristics. The only proteins expressed were PTS and Ribonuclease. In parallel, we expressed CP40 and all were used with ELISA antigen in 49 CLA positive sera and 26 CLA negative sera. The proteins alone showed 100% sensitivity and 96.2%, 92.3%, and 88.5% specificity for rPTS, rRibonuclease, and rCP40, respectively. When proteins were combined, they showed 100% sensitivity and 84.6%, 92.3%, 88.5%, and 92.3% specificity for rPTS/rCp40, rRibonuclease/rCP40, rPTS/rRibonuclease, and rPTS/rRibonuclease/rCP40, respectively. The results of this study show an excellent correlation of sensitivity and specificity with all proteins. None of the specificity values preclude the potential of rPTS, rRibonuclease, or rCP40 for use in ELISA diagnostic assays since the results of this work are superior to those of other studies on CLA diagnosis described in the literature.

Otoliths-composed gelatin/sodium alginate scaffolds for bone regeneration.

Evidence that otoliths, mineral-rich limestone concrescences present in the inner ear of bone fishes, can accelerate bone formation in vivo has been previously reported. The goal of this work was the development, characterization, and evaluation of the cytocompatibility of otoliths-incorporated sodium alginate and gelatin scaffolds. Cynoscion acoupa-derived otoliths were characterized by X-ray fluorescence spectrometry (FRX), particle size, free lime, and weight loss by calcination. Furthermore, otoliths were incorporated into sodium alginate (ALG/OTL-s) or gelatin (GEL/OTL-s) scaffolds, previously developed by freeze-drying. Then, the scaffolds were characterized by thermogravimetric analysis (TGA/DTG), differential scanning calorimetry (DSC), infrared spectroscopy with Fourier transform (FTIR), swelling tests, and scanning electron microscopy (SEM). Cytotoxicity assays were run against J774.G8 macrophages and MC3T3-E1 osteoblasts. Data obtained from TGA/DTG, DSC, and FTIR analyses confirmed the interaction between otoliths and the polymeric scaffolds. SEM showed the homogeneous porous 3D structure rich in otolith micro-fragments in both scaffolds. Swelling of the GEL/OTL-s (63.54 ± 3.0%) was greater than of ALG/OTL-s (13.36 ± 9.9%) (p < 0.001). The viability of J774.G8 macrophages treated with both scaffolds was statistically similar to the group treated with DMEM only (p > 0.05) and significantly higher than that treated with Triton-X (p < 0.01) at 72 h. Both scaffolds showed approximately 100% growth of MC3T3-E1 osteoblasts by 24 h, similarly to control (p > 0.05). However, by 48 h, only ALG/OTL-s showed growth similar to control (p > 0.05), whereas GEL/OTL showed a significantly lower growth index (p < 0.05). In conclusion, the physicochemical profiles suggest proper interaction between the otoliths and the two developed polymeric 3D scaffolds. Moreover, both materials showed cytocompatibility with J774.G8 macrophages but the growth of MC3T3-E1 osteoblasts was higher when exposed to ALG/OTL-s. These data suggest that sodium alginate/otoliths scaffolds are potential biomaterials to be used in bone regeneration applications. Graphical abstract.

The combination of Brazilian red propolis and recombinant protein rCP01850 in the immunoprophylaxis of Corynebacterium pseudotuberculosis infection in mice.

The immunomodulatory properties of Brazilian red propolis (BRP) have been already described. Also, propolis have been proved to have antibacterial activity on Corynebacterium pseudotuberculosis. An adjuvant effect of red propolis oil was able to induce a significant anti-C. pseudotuberculosis humoral immune response. Here, we demonstrate for the first time the immunostimulant property of BRP hydroalcoholic extract (BRPHE) in a recombinant vaccine against caseous lymphadenitis. Mice BALB/c were allocated in three groups inoculated with: sterile saline solution (G1); BRPHE (G2); or BRPHE combined with the C. pseudotuberculosis rCP01850 recombinant protein (G3) in two doses within a 21-days-interval. Blood samples were collected for the total IgG, IgG1 and IgG2a measurement. Mice were challenged with a virulent C. pseudotuberculosis strain, and other 6 mice were used for IFN-γ and IL-10 levels determination after splenocyte stimulation with the recombinant antigen. G3 showed higher significant levels of antibodies on the 42nd experimental day, with a high IgG2a/IgG1 proportion. G2 and G3 presented significant production of IFN-γ and IL-10, while G3 presented the higher levels of IFN-γ (p < 0.05). After challenge, G2 showed a survival rate of 20%, while 70% of mice from G3 survived the experimental challenge. In conclusion, BRPHE used alone has immunostimulant properties specially on cellular immune response, and when used in combination with the recombinant protein rCP01850 induces cellular and humoral immune responses as well as a significant survival of inoculated mice.

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).

Encapsulation of Red Propolis in Polymer Nanoparticles for the Destruction of Pathogenic Biofilms.

Microbial biofilms, structured communities of microorganisms, have been often associated to the infection and bacterial multiresistance problem. Conventional treatment of infection involves the use of antibiotics, being an alternative approach is the use of red propolis, a natural product, to prepare polymer nanoparticles. The aim of the present study was to encapsulate red propolis extract in poly(lactic-co-glycolic acid) (PLGA) nanoparticles for destruction in vitro of pathogenic biofilms. Poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) containing red propolis hydroethanolic extract (2 mg/mL) were produced by emulsification solvent diffusion method. The extract and developed nanoparticles were analyzed for antimicrobial activity and inhibition of bacterial biofilm formation in vitro against Staphylococcus aureus and Pseudomonas aeruginosa. Transmission electron microscopy images confirmed spherical nanoparticles in the range size from 42.4 nm (PLGA NPs) to 69.2 nm (HERP PLGA NPs), with encapsulation efficiencies of 96.99%. The free extract and encapsulated in polymer nanoparticle presented antimicrobial potential, with a minimum inhibitory concentration from 15.6 to 125 µg mL-1 and from 100 to 1560 µg mL-1 to inhibit biofilm formation for the Staphylococcus aureus and Pseudomonas aeruginosa, respectively.

Evaluation of the antioxidant profile and cytotoxic activity of red propolis extracts from different regions of northeastern Brazil obtained by conventional and ultrasound-assisted extraction.

Propolis is a complex mixture of resinous and balsamic material collected from the exudates of plants, shoots, and leaves by bees. This study evaluated red propolis extracts obtained by conventional (ethanolic) extraction and ultrasound-assisted extraction of six samples from different regions of northeastern Brazil. The total phenolic compounds and flavonoids, in vitro antioxidant activity, concentration of formononetin and kaempferol and the cytotoxicity against four human tumor cell lines were determined for all twelve obtained extracts. Significant variations in the levels of the investigated compounds were identified in the red propolis extracts, confirming that the chemical composition varied according to the sampling region. The extraction method used also influenced the resulting propolis compounds. The highest concentration of the compounds of interest and the highest in vitro antioxidant activity were exhibited by the extracts obtained from samples from state of Alagoas. Formononetin and kaempferol were identified in all samples. The highest formononetin concentrations were identified in extracts obtained by ultrasound, thus indicating a greater selectivity for the extraction of this compound by this method. Regarding cytotoxic activity, for the HCT-116 line, all of the extracts showed an inhibition of greater than 90%, whereas for the HL-60 and PC3 lines, the minimum identified was 80%. In general, there was no significant difference (p>0.05) in the antiproliferative potential when comparing the extraction methods. The results showed that the composition of Brazilian red propolis varies significantly depending on the geographical origin and that the method used influences the resulting compounds that are present in propolis. However, regardless of the geographical origin and the extraction method used, all the red propolis samples studied presented great biological potential and high antioxidant activity. Furthermore, the ultrasound-assisted method can be efficiently applied to obtain extracts of red propolis more quickly and with high concentration of biomarkers of interest.

Beer Molecules and Its Sensory and Biological Properties: A Review.

The production and consumption of beer plays a significant role in the social, political, and economic activities of many societies. During brewing fermentation step, many volatile and phenolic compounds are produced. They bring several organoleptic characteristics to beer and also provide an identity for regional producers. In this review, the beer compounds synthesis, and their role in the chemical and sensory properties of craft beers, and potential health benefits are described. This review also describes the importance of fermentation for the brewing process, since alcohol and many volatile esters are produced and metabolized in this step, thus requiring strict control. Phenolic compounds are also present in beer and are important for human health since it was proved that many of them have antitumor and antioxidant activities, which provides valuable data for moderate dietary beer inclusion studies.

Effects of exposure to triphenyltin (TPT) contaminant on sperm activity in adulthood of Calomys laucha exposed through breastfeeding.

Triphenyltin (TPT) is an organotin compound (OT), primarily used in agriculture and in the composition of antifouling paints for ships worldwide. Studies have showed its effects as an endocrine disrupter in several organisms by preventing enzymatic expression and causing reproductive toxicity. This study aimed to evaluate the effects of exposure to TPT, via breastfeeding, on reproductive physiology in the Calomys laucha species. The experimental design was compound of five groups, two controls and three with different doses of TPT. Moreover, females were exposed by gavage to the TPT for 20 days, from the 1st day postpartum to the 21st postnatal day (PND). Then, the pups were euthanized and the kinetics, organelles, and biochemistry of the sperm were evaluated. The results presented a reduction in total motility in the groups exposed to TPT. Regarding cellular organelles analysis, a loss in membrane integrity was evidenced; the functionality of mitochondria showed diminution followed by increased acrosome reaction. In conclusion, the TPT causes alteration of the reproductive parameters, decreasing the activity and sperm quality in individuals exposed in the breastfeeding phase.

Induction and growth curve of calli from leaf and nodal explants of genipap / Indução e curva de crescimento de calos obtidos de explantes foliares e nodais de jenipapeiro

The aim of this study was to determine the effect of the auxin 2,4-D (2,4-dichlorophenoxyacetic) in calli formation from leaf and nodal segments of genipap and to characterize its growth curve. Explants obtained from shoots previously established from in vitro seedlings were used for calli induction. The experimental design was completely randomized in a 3x5x2 factorial with three accessions (NB, SA, SAL), five concentrations of 2,4-D (0.0; 2.0; 4.0, 6.0 or 8.0 mg L-1) and two times of measurement for calli fresh weight (30 and 60 days). There was callus formation in all treatments tested. It was observed that the best response for callus induction from leaf segments was with 2.0 mg L-1 of 2,4-D. For the nodal segment, the response among the accessions was different due to 2,4-D concentrations. The growth curve was plotted according to the fresh weight of callus obtained at intervals of 10 days up to 60 days. Through the established growth curve, the nodal-derived calli from accession SA should be transferred to a new medium, after 40 days of culture.

O objetivo desse trabalho foi determinar o efeito da auxina 2,4-D (ácido diclorofenoxiacético) na calogênese de segmentos foliar e nodal de jenipapeiro e caracterizar sua curva de crescimento. Explantes obtidos de brotações pré-estabelecidas a partir de plântulas in vitro foram utilizados na indução de calos. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 3x5x2, com três acessos (NB, SA e SAL), cinco concentrações de 2,4-D (0,0; 2,0; 4,0; 6,0 ou 8,0 mg L-1) e dois tempos de avaliação (30 e 60 dias) da massa fresca de calos. Houve formação de calos em todos os tratamentos testados. Observou-se que a melhor resposta de indução ocorreu na concentração de 2,0 mg L-1 para calos oriundos de segmentos foliares. Para o segmento nodal a resposta entre os acessos foi diferenciada em função das concentrações de 2,4-D. A curva de crescimento foi plotada a partir da massa fresca dos calos obtida em intervalos de 10 dias até os 60 dias. Através da curva de crescimento estabelecida, os calos derivados de segmentos nodais do acesso SA devem ser transferidos para um novo meio de cultura, 40 dias após à inoculação.

Immune-Informatic Analysis and Design of Peptide Vaccine From Multi-epitopes Against Corynebacterium pseudotuberculosis.

Caseous lymphadenitis (CLA) is a disease caused by Corynebacterium pseudotuberculosis bacteria that affects sheep and goats. The absence of a serologic diagnose is a factor that contributes for the disease dissemination, and due to the formation of granuloma, the treatment is very expensive. Therefore, prophylaxis is the approach with best cost-benefit relation; however, it still lacks an effective vaccine. In this sense, this work seeks to apply bioinformatic tools to design an effective vaccine against CLA, using CP40 protein as standard for the design of immunodominant epitopes, from which a total of 6 sequences were obtained, varying from 10 to 16 amino acid residues. The evaluation of different properties of the vaccines showed that the vaccine is a potent and nonallergenic antigen remaining stable in a wide range of temperatures. The initial tertiary structure of the vaccine was then predicted and a model selected. Later, the process of CP40 protein and TLR2 receptor binding was performed, presenting interaction with this receptor, which plays an important role in the activation of the immune response.

Photocatalytic and Cytotoxic Effects of Nitrogen-Doped TiO2 Nanoparticles on Melanoma Cells.

Titanium dioxide (TiO2) has attracted attention as a photosensitizer in the field of photodynamic therapy (PDT) due to its low toxicity and high photostability. In the present work, nitrogen-doped TiO2 (N-TiO2) nanoparticles had their photokilling efficiency evaluated on a murine melanoma cell line (B16-F10) and fibroblasts (NIH 3T3). The N-TiO2 nanoparticles were prepared by a modified hydrogen peroxide sol-gel process using triethylamine as nitrogen precursor. XRD measurements showed that all TiO2 and N-TiO2 samples consisted of an anatase crystalline phase and no trace of rutile was detected. N-TiO2 nanoparticles showed higher absorbance in the visible region than pure TiO2. Nanoparticle dosage increase from 0.1 mg/ml to 0.5 mg/ml played a role in cell viability, causing high cytotoxicity in melanoma and fibroblast cells. The cytotoxic potential of N-TiO2 on cells was analyzed using visible light, UV-A irradiation and dark conditions. All samples were cytotoxic in a PDT test, and N-TiO2 caused 93% death in melanoma cells under UV irradiation treatment at 0.5 mg/ml. Gene expression analysis of this sample showed, under ultraviolet photoexcitation, an increase of pro-apoptotic BAX gene expression, suggesting cell death by apoptosis.

Antitumor activity of Brazilian red propolis fractions against Hep-2 cancer cell line.

Continuous increases in the rates of tumor diseases have highlighted the need for identification of novel and inexpensive antitumor agents from natural sources. In this study, we investigated the effects of enriched fraction from hydroalcoholic Brazilian red propolis extract against Hep-2 cancer cell line. Initially 201 fractions were arranged in 12 groups according to their chromatographic characteristics (A-L). After an in vitro cell viability screening, J and L were further selected as promising enriched fractions for this study. The chemical characterization was performed and Biochanin A, Formononetin, and Liquiritigenin compounds were quantified. Through MTT viability assay and morphological changes observed by Giemsa and DAPI staining, the results showed that red propolis inhibited cancer cells growth. Flow cytometry results indicated effects that were partly mediated through programmed cell death as confirmed by externalization of phosphatidylserine, DNA cleaved assay, increase at SUB G1-G0 phase in cell cycle analysis and loss of mitochondrial membrane potential. In conclusion, our results demonstrated that red propolis enriched fractions promoted apoptotic effects in human cancer cells through the mechanisms involving mitochondrial perturbation. Therefore, red propolis fractions contain candidate agents for adjuvant cancer treatment, which further studies should elucidate the comprehensive mechanistic pathways.

Effect of Tamarindus indica L. and Manihot esculenta Extracts on Antibiotic-resistant Bacteria.

BACKGROUND: The chemical composition of plants used in traditional medicine exhibits biologically active compounds, such as tannins, flavonoids, and alkaloids and becomes a promising approach to treat microbial infections, mainly with drug-resistant bacteria. OBJECTIVE: The aim of the present study was to evaluate the hydroethanolic leaf extracts of Tamarindus indica (tamarind) and Manihot esculenta (cassava) as antimicrobial potential against Pseudomonas aeruginosa clinical isolated and Methicillin-resistant Staphylococcus aureus. MATERIALS AND METHODS: Hydroethanolic leaf extracts were prepared and characterized by high-performance liquid chromatography/diode array detection, Fourier transform infrared, 1,1-diphenyl-2-picrylhydrazyl, and ultraviolet-visible methods. The antimicrobial activity against four strains of clinical relevance was evaluated by the microdilution method at minimum inhibitory concentrations. RESULTS: Phenolic compounds such as flavonoids were detected in the plant extracts. T. indica extract at 500 µg/mL showed antimicrobial activity against S. aureus and P. aeruginosa; however, M. esculenta showed only activity against P. aeruginosa in this concentration. CONCLUSIONS: Our results suggested that polyphenols and flavonoids present in T. indica leaf extracts are a potential source of antimicrobial compound. The T. indica extract showed antibacterial activity against S. aureus and P. aeruginosa while M. esculenta had effect only on P. aeruginosa meropenem resistant. SUMMARY: Antibacterial effect of T. indica and M. esculenta leaf extract was evaluated.T. indica extract displayed activity against S. aureus and P. aeruginosa strains.M. esculenta showed effect on P. aeruginosa meropenem resistant. Abbreviations Used: BHI: Agar brain heart infusion, CAPES: Coordination for the improvement of higher education personnel, DPPH: 1,1-diphenyl-2-picrylhydrazyl, FAPITEC/SE: Foundation for support to research and technological innovation of the state of sergipe, FTIR: Fourier transform infrared spectroscopy, HPLC: High-performance liquid chromatography, KBr: Potassium bromide, MIC: Minimum inhibitory concentration, MRSA: Methicillin-resistant Staphylococcus aureus, RSC: Radical scavenging capacity, UV-vis: Ultraviolet-visible.

Antioxidant, antimicrobial, antiparasitic, and cytotoxic properties of various Brazilian propolis extracts.

Propolis is known for its biological properties and its preparations have been continuously investigated in an attempt to solve the problem of their standardization, an issue that limits the use of propolis in food and pharmaceutical industries. The aim of this study was to evaluate in vitro antioxidant, antimicrobial, antiparasitic, and cytotoxic effects of extracts of red, green, and brown propolis from different regions of Brazil, obtained by ethanolic and supercritical extraction methods. We found that propolis extracts obtained by both these methods showed concentration-dependent antioxidant activity. The extracts obtained by ethanolic extraction showed higher antioxidant activity than that shown by the extracts obtained by supercritical extraction. Ethanolic extracts of red propolis exhibited up to 98% of the maximum antioxidant activity at the highest extract concentration. Red propolis extracts obtained by ethanolic and supercritical methods showed the highest levels of antimicrobial activity against several bacteria. Most extracts demonstrated antimicrobial activity against Staphylococcus aureus. None of the extracts analyzed showed activity against Escherichia coli or Candida albicans. An inhibitory effect of all tested ethanolic extracts on the growth of Trypanosoma cruzi Y strain epimastigotes was observed in the first 24 h. However, after 96 h, a persistent inhibitory effect was detected only for red propolis samples. Only ethanolic extracts of red propolis samples R01Et.B2 and R02Et.B2 showed a cytotoxic effect against all four cancer cell lines tested (HL-60, HCT-116, OVCAR-8, and SF-295), indicating that red propolis extracts have great cytotoxic potential. The biological effects of ethanolic extracts of red propolis revealed in the present study suggest that red propolis can be a potential alternative therapeutic treatment against Chagas disease and some types of cancer, although high activity of red propolis in vitro needs to be confirmed by future in vivo investigations.

Chemical Composition and Biological Activity of Extracts Obtained by Supercritical Extraction and Ethanolic Extraction of Brown, Green and Red Propolis Derived from Different Geographic Regions in Brazil.

The variations in the chemical composition, and consequently, on the biological activity of the propolis, are associated with its type and geographic origin. Considering this fact, this study evaluated propolis extracts obtained by supercritical extraction (SCO2) and ethanolic extraction (EtOH), in eight samples of different types of propolis (red, green and brown), collected from different regions in Brazil. The content of phenolic compounds, flavonoids, in vitro antioxidant activity (DPPH and ABTS), Artepillin C, p-coumaric acid and antimicrobial activity against two bacteria were determined for all extracts. For the EtOH extracts, the anti-proliferative activity regarding the cell lines of B16F10, were also evaluated. Amongst the samples evaluated, the red propolis from the Brazilian Northeast (states of Sergipe and Alagoas) showed the higher biological potential, as well as the larger content of antioxidant compounds. The best results were shown for the extracts obtained through the conventional extraction method (EtOH). However, the highest concentrations of Artepillin C and p-coumaric acid were identified in the extracts from SCO2, indicating a higher selectivity for the extraction of these compounds. It was verified that the composition and biological activity of the Brazilian propolis vary significantly, depending on the type of sample and geographical area of collection.

Hydroalcoholic extract of red propolis promotes functional recovery and axon repair after sciatic nerve injury in rats.

CONTEXT: Peripheral axon injury and degeneration are often mediated by oxidative stress and inflammation. The hydroalcoholic extract of the red propolis (HERP) has attracted great attention because of its antioxidant and anti-inflammatory activities. OBJECTIVE: The objective of this work is to study the effect of HERP on nerve repair and functional recovery after sciatic nerve injury (SNI) in rats. MATERIALS AND METHODS: The chemical markers in HERP were identified using high-resolution mass spectroscopy. After axonotmesis of sciatic nerve, ibuprofen (IBP) and HERP treatments were orally administered for 28 d. Behavioural tests were performed weekly after SNI. The myelinated axon number was counted using morphometric analysis. RESULTS: The compounds found in HERP were pinocembrin, formononetin, vestitol, and biochanin A. The animals that underwent SNI showed a significant decrease in motor function based on the Basso, Beattie and Bresnahan scale and sciatic functional index compared with sham animals until 7 d after the surgery (p < 0.05). After 14 and 21 d, the SNI groups treated with either HERP or IBP showed significant improvement (p < 0.01), and the SNI group treated with HERP 10 mg/kg showed accelerated motor recovery compared with the other groups (p < 0.01). SNI caused also a reduction in the myelinated axon counts, and treatment with HERP 10 mg/kg induced a significant increase in the number of myelinated fibres compared with all other groups. CONCLUSION: HERP promoted regenerative responses and accelerated functional recovery after sciatic nerve crush. Thus, it can be considered to be a new strategy or complementary therapy for treating nerve injuries.

Determination of Parameters for the Supercritical Extraction of Antioxidant Compounds from Green Propolis Using Carbon Dioxide and Ethanol as Co-Solvent.

The aim of this study was to determine the best processing conditions to extract Brazilian green propolis using a supercritical extraction technology. For this purpose, the influence of different parameters was evaluated such as S/F (solvent mass in relation to solute mass), percentage of co-solvent (1 and 2% ethanol), temperature (40 and 50°C) and pressure (250, 350 and 400 bar) using supercritical carbon dioxide. The Global Yield Isotherms (GYIs) were obtained through the evaluation of the yield, and the chemical composition of the extracts was also obtained in relation to the total phenolic compounds, flavonoids, antioxidant activity and 3,5-diprenyl-4-hydroxicinnamic acid (Artepillin C) and acid 4-hydroxycinnamic (p-coumaric acid). The best results were identified at 50°C, 350 bar, 1% ethanol (co-solvent) and S/F of 110. These conditions, a content of 8.93±0.01 and 0.40±0.05 g/100 g of Artepillin C and p-coumaric acid, respectively, were identified indicating the efficiency of the extraction process. Despite of low yield of the process, the extracts obtained had high contents of relevant compounds, proving the viability of the process to obtain green propolis extracts with important biological applications due to the extracts composition.