Therapeutic Potential of Bioactive Compounds from Brugmansia suaveolens Bercht. & J. Presl.

Brugmansia suaveolens Bercht. & J. Presl has been widely used due to the presence of different bioactive compounds. This review summarizes the latest advances and perspectives of the B. suaveolens plant species; it is a systematic literature review on aspects of botany, traditional uses, phytochemistry, pharmacology, and toxicology as therapeutic potential. In addition, 120 compounds are described, including alkaloids, flavonoids, terpenoids, steroids, amino acids, aromatics, and aliphatics. As for the therapeutic potential, it is described in extracts and compounds in the antitumor, anti-inflammatory, antioxidant, antimicrobial, antispasmodic, anticoagulant, and analgesic aspects, as well as the effects on the central nervous system. The toxicity of the genus stands out, especially the potential for organ toxicity. Therefore, this review evidenced the knowledge related to the traditional use based on the scientific research of Brugmansia suaveolens, highlighting an overview of bioactive compounds and biological and toxicological activities in order to provide a scientific basis for future studies on the value of this species for the development of new natural products.

Phytochemical analysis of Brugmansia suaveolens Bercht. & J. Presl and its therapeutic potential for topical use.

Brugmansia suaveolens Bercht. & J. Presl represents a promising source of new active molecules. Therefore, the aim of the study is to outline the profile of secondary metabolites and their therapeutic potential and in vitro safety properties. The identification of substances was carried out through the chromatographic profile, while the evaluation of therapeutic use was conducted through in vitro biological assays of antioxidant and antimicrobial activity and quantification of the total phenolic content. The safety of the extracts was evaluated using a cytotoxicity assay. The results found revealed the presence of different secondary metabolites, such as flavonoids and alkaloids. Biological assays showed promising antimicrobial activity in gram-negative strains. Regarding safety, greater cytotoxicity is observed in macrophage cells. The study demonstrated that the extracts are potent for therapeutic use, aiming at the development of a phytoproduct for topical use, providing an innovative, relevant and significant character for future research.

Dentine biomodification by sulphonamides pre-treatment: bond strength, proteolytic inhibition, and antimicrobial activity.

We evaluated the effects of dentine biomodification after pre-treatment with two sulphonamide carbonic anhydrase inhibitors (CAIs) of the N-[4-sulphamoylphenethylcarbamoyl]benzenesulphonamide type, investigating matrix metalloproteases activity, resin-dentine micro tensile bond strength, dentine surface wettability, and antimicrobial activities. Ninety-five sound-extracted human molars were selected for the study. Inhibitory effects were evaluated by gelatinase and collagenase activity tests and collagen degradation FT-IR spectroscopic analysis. Pre-treatment with the two CAIs kept the micro tensile values after 12 months of storage (32.23 ± 5.95) and cariogenic challenge (34.13 ± 2.71) similar to the initial, pre-treatment values (33.56 ± 4.34). A decreased Streptococcus mutans biofilm formation on dentine surfaces and antibacterial activity against planktonic bacteria were observed after CAI treatment. Dentine pre-treatment with sulphonamide CAIs maintained adhesion strength stability, allowed better dentine wettability, maintained matrix collagen, and showed anti-S. mutans activity.

Chagas Disease: Perspectives on the Past and Present and Challenges in Drug Discovery.

Chagas disease still has no effective treatment option for all of its phases despite being discovered more than 100 years ago. The development of commercial drugs has been stagnating since the 1960s, a fact that sheds light on the question of how drug discovery research has progressed and taken advantage of technological advances. Could it be that technological advances have not yet been sufficient to resolve this issue or is there a lack of protocol, validation and standardization of the data generated by different research teams? This work presents an overview of commercial drugs and those that have been evaluated in studies and clinical trials so far. A brief review is made of recent target-based and phenotypic studies based on the search for molecules with anti-Trypanosoma cruzi action. It also discusses how proteochemometric (PCM) modeling and microcrystal electron diffraction (MicroED) can help in the case of the lack of a 3D protein structure; more specifically, Trypanosoma cruzi carbonic anhydrase.

Quantification of schizophyllan directly from the fermented broth by ATR-FTIR and PLS regression.

Non-destructive methods that allow the quantification of bioproducts in a simple and quick manner during fermentation are extremely desirable from a practical point of view. Therefore, a 9 day fermentation experiment with Schizophyllum commune was carried out to investigate the possibility of using ATR-FTIR to quantify the schizophyllan biopolymer (SPG) directly from the culture medium. On each day, aliquots of the fermentation were taken, and the cell-free supernatant was analyzed by ATR-FTIR. The main objective of this step was to evaluate whether FTIR would be able to detect the appearance of specific peaks related to the production of SPG. The results of the PCA analysis showed that there was a reasonable separation of the days through the FTIR spectra. Then PCA-LDA was applied to the same dataset, which confirmed the formation of groups for each day of fermentation, after which, a calibration and test set was developed. Through a matrix generated by an experimental design with 2 factors and 5 levels, 25 samples were created with variations in the concentration of the culture medium and SPG. The ATR-FTIR spectra of this data set were modeled using PLS regression with backward selection of predictors. The results revealed that the amount of SPG produced can be quantified directly in the culture medium with excellent precision with R2CV = 0.951, R2P = 0.970, RMECV = 0.205 g, RMSEP = 0.170 g, RPDcv = 4.53 and RPDp = 5.88. The traditional method to quantify SPG is time consuming, requires several steps and uses solvents. In contrast, the method proposed in this work is a viable, faster, and a simpler alternative, which does not use reagents and does not require extensive pre-treatment of the samples.

New method for rapid identification and quantification of fungal biomass using ergosterol autofluorescence.

This research reports on the development of a method to identify and quantify fungal biomass based on ergosterol autofluorescence using excitation-emission matrix (EEM) measurements. In the first stage of this work, several ergosterol extraction methods were evaluated by APCI-MS, where the ultrasound-assisted procedure showed the best results. Following an experimental design, various quantities of the dried mycelium of the fungus Schizophyllum commune were mixed with the starchy solid residue (BBR) from the babassu (Orbignya sp.) oil industry, and these samples were subjected to several ergosterol extraction methods. The EEM spectral data of the samples were subjected to Principal Component Analysis (PCA), which showed the possibility to qualitatively evaluate the presence of ergosterol in the samples by ergosterol autofluorescence without the addition of any reagent. In order to assess the feasibility of quantifying fungal biomass using ergosterol autofluorescence, the EEM spectral data and known amounts of fungal biomass were modeled using partial least squares (PLS) regression and a procedure of backward selection of predictors (AutoPLS) was applied to select the Excitation-Emission wavelength pairs that provide the lowest prediction error. The results revealed that the amount of fungal biomass in samples containing interfering substances (BBR) can be accurately predicted with R2CV = 0.939, R2P = 0.936, RPDcv = 4.07, RPDp = 4.06, RMSECV = 0.0731 and RMSEP = 0.0797. In order to obtain an easy-to-understand equation that expresses the relationship between fungal biomass and fluorescence intensity, multiple linear regression (MLR) was applied to the VIP variables selected by the AutoPLS method. The MLR model selected only 2 variables and showed a very good performance, with R2CV = 0.862, R2P = 0.809, RPDcv = 2.18, RPDp = 2.35, RMSECV = 0.137 and RMSEP = 0.138. This study demonstrated that ergosterol autofluorescence can be successfully used to quantify fungal biomass even when mixed with agroindustrial residues, in this case BBR.

Carotenoids from UV-resistant Antarctic Microbacterium sp. LEMMJ01.

The Microbacterium sp. LEMMJ01 isolated from Antarctic soil does not belong to any of the nearest species identified in the RDP database. Under UV radiation (A, B and C wavebands) the survival fractions of Microbacterium sp. cells were much higher compared with wild-type E. coli K12A15. Especially remarkable for an Antarctic bacterium, an expressive resistance against high UV-B doses was observed. The increased survival of DNA repair-proficient E. coli grown overnight added of 0.1 mg/ml or 1 mg/ml of the whole pigment extract produced by Microbacterium sp. revealed that part of the resistance of Microbacterium sp. against UV-B radiation seems to be connected with photoprotection by its pigments. Scanning electron microscopy revealed that UV-A and UV-B ensued membrane alterations only in E. coli. The APCI-MS fingerprints revealed the diagnostic ions for neurosporene (m/z 580, 566, 522, 538, and 524) synergism for the first time in this bacterium by HPLC-MS/MS analysis. Carotenoids also were devoid of phototoxicity and cytotoxicity effects in mouse cells and in human keratinocytes and fibroblasts.

Clove oil nanoemulsion showed potent inhibitory effect against Candida spp.

Increasing resistance to current fungicides is a clinical problem that leads to the need for new treatment strategies. Clove oil (CO) has already been described as having antifungal action. However, it should not be applied directly to the skin as it may be irritating. One option for CO delivery and suitable topical application would be nanoemulsions (NEs). NEs have advantages such as decreased irritant effects and lower dose use. The purpose of this work was the development of NEs containing CO and in vitro evaluation against Candida albicans and Candida glabrata. The NEs were produced by an ultrasonic processor with different proportions of CO and Pluronic® F-127. In order to determine the best composition and ultrasound amplitude, an experimental design was performed. For the evaluation, droplet size and polydispersity index (PdI) were used. After the stability study, in vitro activity against C. albicans and C. glabrata was evaluated. NEs selected for the stability study, with diameter <40 nm and PdI <0.2, remained stable for 420 d. Activity against Candida spp. was improved when the CO was nanoemulsified, for it possibly leads to a better interaction between the active and the microorganisms, mainly in C. albicans.

Evaluation of Kluyveromyces marxianus endo-polygalacturonase activity through ATR-FTIR.

The endo-polygalacturonase enzyme (endoPG: EC 3.2.1.15) plays an important role in the fruit juice and wine industries, so the development of new tools for the quantitative and qualitative analysis of its enzymatic action is necessary. In this work, we report the development of a simple, fast and practical method that did not use any chemical reagent to identify and evaluate the action of the endoPG enzyme, produced by the yeast Kluyveromyces marxianus CCT3172, using attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy combined with principal component analysis-linear discriminant analysis (PCA-LDA). This method evaluated the action of the endoPG enzyme on the polygalacturonic acid (PGA) substrate at 5 different times (0, 10, 15, 20 and 30 minutes), and at each time interval the samples were analyzed by ATR-FTIR. It was demonstrated that there was clear segregation between the samples that were and that were not subjected to the action of the endoPG enzyme, and it was also possible to distinguish the samples that were subjected to different incubation times with the enzyme. Through PCA-LDA it was possible to obtain wavelengths that are biomarkers for this enzymatic reaction and the observed changes as a function of hydrolysis duration were found to be in agreement with the breakdown of the glycosidic chain (1011 cm-1-CH-O- CH stretching) of PGA and release of oligosaccharides (1078 cm-1 C-OH elongation). The activity of the endoPG enzyme and the release of galacturonic acid were verified by the dinitrosalicylic acid (DNS) method in all samples. The efficacy of an automatic classifier using a principal component analysis-linear discriminant classifier (PCA-LDC) was evaluated to diagnose the action of the endoPG enzyme. The results showed an accuracy of 100% for the identification of the endoPG enzyme action and from 91.67% to 100% for classification according to the hydrolysis duration in which PGA was exposed to endoPG. The present study indicates that this methodology may be a new approach for the qualitative evaluation of the endoPG enzyme with the potential to be used in laboratories and industries.

Development and characterization of a nanoemulsion containing propranolol for topical delivery.

BACKGROUND: Propranolol (PPN) is a therapeutic option for the treatment of infantile hemangiomas. This study aimed at the development of nanoemulsion (NE) containing 1% PPN, characterization of the system, and safety studies based on ex vivo permeation, cytotoxicity, and biodistribution in vivo. METHODS: The formulation was developed and characterized in relation to the droplet size, polydispersity index (PDI), pH, zeta potential, and electronic microscopy. Ex vivo permeation studies were used to evaluate the cutaneous retention of PPN in the epidermis and dermis. Cytotoxicity studies were performed in fibroblasts, macrophages, and keratinocytes. In vivo biodistribution assay of the formulations was performed by means of labeling with technetium-99m. RESULTS: NE1 exhibited droplet size of 26 nm, PDI <0.4, pH compatible with the skin, and zeta potential of -20 mV, which possibly contributes to the stability. Electron microscopy showed that the NE presented droplets of nanometric size and spherical shape. NE1 provided excellent stability for PPN. In the ex vivo cutaneous permeation assay, the NE provided satisfactory PPN retention particularly in the dermis, which is the site of drug action. In addition, NE1 promoted cutaneous permeation of the PPN in small amount. In vivo biodistribution showed that the radiolabeled formulation remained in the skin and a small amount reached the bloodstream. NE1 presented low cytotoxicity to fibroblasts, macrophages, and keratinocytes in the concentrations evaluated in the cytotoxicity assay. CONCLUSION: We concluded that the formulation is safe for skin administration; however, cutaneous irritation studies should be performed to confirm the safety of the formulation before clinical studies in patients with infantile hemangiomas.

Administração oral de piperina em frangos de corte / Piperine oral administration on broiler' chikens

O efeito tóxico da piperina, principal amida presente em diversas espécies de pimenta, foi avaliada em frangos de corte por meio da administração oral (0,00, 1,12, 2,25 e 4,5mg kg-1 peso vivo) por 14 dias consecutivos. Foram empregados 60 pintos machos (Cobb Avian 48) com sete dias de idade, distribuídos aleatoriamente em quatro grupos experimentais (n=15). Foram avaliados parâmetros como: ganho de peso, peso relativo do fígado e alterações hematológicas e anatomopatológicas. A administração oral de piperina não interferiu no ganho de peso ou no peso relativo do fígado, além de não promover alteração no tamanho e na coloração dos órgãos ou no aparecimento de lesões de parênquima e nas mucosas do órgão. Todavia, alterações histopatológicas dose-dependentes foram observadas. A piperina não foi capaz de alterar significativamente os parâmetros hematológicos analisados, com exceção do leucograma, em que as doses de 1,12mg e 2,25mg kg-1 promoveram aumento do número de heterófilos e do número total de leucócitos, respectivamente. Os resultados sugerem que a dose oral de 1,12mg de piperina por quilo não é tóxica para frangos de corte, sendo semelhante aos resultados obtidos para ratos e camundongos. Além disso, constatou-se a capacidade da piperina em aumentar o número total de leucócitos circulantes a partir da dose de 2,25mg kg-1 nessa espécie animal.

The toxic effect of piperine, the main amide compound of different pepper species, was evaluated on broiler chickens by oral administration at 0.00, 1.12, 2.25 and 4.50mg kg-1 concentrations for 14 days. Sixty seven days old male chicks (Cobb Avian 48) randomly allocated to four experimental groups (n=15) were used in this work. Parameters such as: body weight gain, liver relative weight, hematological and anatomopathological alterations were analyzed. The oral route administration did not interfere on weight gain or liver relative weight, as well as, any modification on size and organs' color and/or parenchyma/mucous membranes injuries were observed; however, liver histopathological changes were noticed in a dose-dependent manner. In addition, piperine did not alter hematological parameters, except for leukocytes counting, which at 1.12 and from 2.25mg kg-1 caused an increase of heterophils and in the total number of leukocytes, respectively. The results suggest that 1.12mg kg-1 of piperine orally administrated is not toxic for broiler chickens, as previously observed for rats and mice. Moreover, 2.25mg kg-1 of piperine seems to increase the total number of leukocytes for this animal specie.

A serological study on Brucella abortus, caprine arthritis-encephalitis virus and Leptospira in dairy goats in Rio de Janeiro, Brazil.

In spite of the large number of goats found in several developing tropical countries, milk production remains unsatisfactory. The occurrence of infectious diseases, such as leptospirosis, brucellosis and caprine arthritis-encephalitis (CAE) may in part be responsible for sub-optimal production. In this study, 1000 serum samples were tested for leptospirosis, 953 for brucellosis and 562 for CAE. All tested flocks presented at least one seroreactive animal for leptospirosis and for CAE. Reactivity to leptospirosis was 11.1%, and serovar hardjo was the most frequently found. Anti-B. abortus agglutinins were found in 0.5% of the samples presented and 14.1% were seroreactive to CAE. Leptospirosis was considered to represent the major infectious problem in the studied goat flocks. The occurrence of infectious diseases in the tested flocks may represent an important factor contributing to the decreased productivity of the animals. These findings may be similar to those observed in other developing countries and require further study to define the relationship between seropositivity and reduced production.