Molecular docking and evaluation of antileishmania activity of a ruthenium complex with epiisopiloturine and nitric oxide

Leishmaniasis is an infectious disease that affects both animals and humans, caused by flagellated parasites belonging to the genus Leishmania. The disease is estimated to reach about 700,000 to 1 million people, causing the deaths of 20 to 30,000 individuals annually. Thus, the present study aims to perform molecular docking tests and evaluation of antileishmania activity in vitro of a ruthenium complex with epiisopiloturine and nitric oxide. AutoDockTools-1.5.6 software was used to perform molecular docking tests. Molecular targets were considered rigid, and Epiruno2 considered flexible. The genetic algorithm Lamarckian (AGL) with global search and pseudo-Solis and Wets with local search were the methods adopted in the docking. The most promising results of molecular interaction were achieved in the targets Pteridine reductase and UDP-glucose Pyrophosphorylase with rates of -10.68 Kcal·mol-1 and -10.51 Kcal·mol-1, respectively. This demonstrates that Epiruno2 has molecular affinity with the targets of L. major. In vitro assays prove the antileishmania activity of the complex in the face of promastigote forms with inhibition of growth, concluding through this study that the Epiruno2 complex has antileishmania activity

Evaluation of the serum biochemistry and histopathology of kidney and bladder of dogs with Leishmania sp. in their urine

The visceral establishment of Leishmania infantum in dogs may result in kidney and bladder tissue injury, with L. infantum ending up in urine. This study therefore aimed at investigating the presence of Leishmania sp. in urinary sediments, and correlating the results with those from renal and bladder serum biochemistry and histopathology. Thirty dogs with negative Nested-Polymerase Chain Reaction (PCR) for E. canis were used in the experiment, and were divided into three groups: control group (10 dogs), neither leishmaniasis nor clinical changes; group I (15 dogs), leishmaniasis but no Leishmania sp. in urine; and group II (5 dogs), leishmaniasis, as well as Leishmania sp. in urine. All animals were submitted to clinical, serological, and parasitological diagnosis for leishmaniasis, biochemical exams, and kidney and bladder histopathology. The parasite was also detected in the bladder imprint of one group II dog. Group II dogs presented with very low albumin concentrations, low albumin/globulin ratios, and kidney and bladder lesions. In the kidneys, hydropic degeneration, thickened Bowman's capsule, and thickening of the tubular capsule were detected in all dogs with positive urinary sediment. However, no significant difference in these renal changes was observed between groups. The intensity and distribution of bladder inflammatory infiltrates were significantly (p-value < 0.05, Kruskal-Wallis’ and Dunn’s tests) higher in group II dogs, compared with those of the other groups. The presence of Leishmania sp. in the urine of infected dogs appeared to be related to low serum albumin concentrations and more severe bladder lesions

Platonia insignis

Platonia insignis Mart., popularly known as “bacurizeiro,” is used in traditional medical practices based on its diverse biological properties. This study was aimed at evaluating the antileishmanial effects of the ethanol extract (EtOH-Ext), hexane fraction (Hex-F), and its main isolated Lupeol obtained from stem barks of P. insignis against Leishmania (Leishmania) amazonensis, as well as their cytotoxicity and possible mechanisms of action. The EtOH-Ext, Hex-F, and Lupeol inhibited the growth of L. amazonensis promastigote forms at IC50 of 174.24, 45.23, and 39.06 µg/mL, respectively, as well as L. amazonensis axenic amastigote forms at IC50 of 40.58, 35.87, and 44.10 µg/mL, respectively. The mean cytotoxic concentrations for macrophages (CC50) were higher than those for amastigotes (341.95, 71.65, and 144.0 µg/mL, resp.), indicating a selective cytotoxicity towards the parasite rather than the macrophages. Interestingly, all treatments promoted antileishmanial effect against macrophage-internalized amastigotes at concentrations lower than CC50. Furthermore, increases of lysosomal volume of macrophages treated with EtOH-Ext, Hex-F, and Lupeol were observed. On the other hand, only Lupeol stimulated increase of phagocytic capability of macrophages, suggesting this compound might be characterized as the biomarker for the antileishmanial effect of P. insignis stem bark, as well as the involvement of immunomodulatory mechanisms in this effect

Copper nanoparticles stabilized with cashew gum: Antimicrobial activity and cytotoxicity against 4T1 mouse mammary tumor cell line

Copper nanoparticles stabilized with cashew (CG-CuNPs) were synthesized by reduction reaction using ascorbic acid and sodium borohydride, using the cashew gum (CG) as a natural polymer stabilizer. Dynamic light scattering, atomic force microscopy, Fourier-transform infrared spectroscopy, UV-Vis spectrophotometry, and x-ray diffraction were used to characterize the nanoparticles (CG-CuNPs), and copper was quantified by electrochemical measurement. The UV-vis spectra of the CG-CuNPs confirmed the formation of nanoparticles by appearance of a surface plasmon band at 580 nm after 24 h of reaction. The Fourier-transform infrared spectrum of CG-CuNPs showed the peak at 1704 cm−1 from cashew gum, confirming the presence of the gum in the nanoparticles. The average size of CG-CuNPs by dynamic light scattering and atomic force microscopy was around 10 nm, indicating small, approximately spherical particles. Antimicrobial assays showed that CG-CuNPs had activity against Staphylococcus aureus ATCC 29213 with a minimal inhibitory concentration of 0.64 mM. The cytotoxicity assay on BALB/c murine macrophages showed lower cytotoxic effects for CG-CuNPs than CuSO4·5H2O. Viability cell assays for CG-CuNPs at (0.250 mM) inhibited by 70% the growth of 4T1 LUC (4T1 mouse mammary tumor cell line) and NIH 3T3 cells (murine fibroblast cells) over a 24-h period. Therefore, CG-CuNPs can be used as an antimicrobial agent with lower cytotoxic effects than the CuSO4·5H2O precursor.The author would like to thank at UCM for performingDPV, USP by X-ray diffraction experiment, REQUIMTE/LAQV for FTIR, UnB and UFPI for the cytotoxicityassays, as well as at UFPI for help with DLS, UV-Vis,AFM, and microbiological experiments. This work was supported by Project 400398/2014-1—Desenvolvimento de Nanopartículas Estabilizadas com Goma de Cajueiro para Aplicações Biotecnologicas, financed by CNPq. AlexandraPlácido is grateful to FCT by her grant SFRH/BD/97995/2013, financed by POPH-QREN-Tipologia 4.1-Formação Avançada, subsidized by Fundo Social Europeu and Ministério da Ciência, Tecnologia e Ensino Superior.info:eu-repo/semantics/publishedVersio

Behavior and biocompatibility of rabbit bone marrow mesenchymal stem cells with bacterial cellulose membrane

Background Tissue engineering has been shown to exhibit great potential for the creation of biomaterials capable of developing into functional tissues. Cellular expansion and integration depends on the quality and surface-determinant factors of the scaffold, which are required for successful biological implants. The objective of this research was to characterize and evaluate the in vitro characteristics of rabbit bone marrow mesenchymal stem cells (BM-MSCs) associated with a bacterial cellulose membrane (BCM). We assessed the adhesion, expansion, and integration of the biomaterial as well as its ability to induce macrophage activation. Finally, we evaluated the cytotoxicity and toxicity of the BCM. Methods Samples of rabbit bone marrow were collected. Mesenchymal stem cells were isolated from medullary aspirates to establish fibroblast colony-forming unit assay. Osteogenic, chondrogenic, and adipogenic differentiation was performed. Integration with the BCM was assessed by scanning electron microscopy at 1, 7, and 14 days. Cytotoxicity was assessed via the production of nitric oxide, and BCM toxicity was assessed with the MTT assay; phagocytic activity was also determined. Results The fibroblastoid colony-forming unit (CFU-F) assay showed cells with a fibroblastoid morphology organized into colonies, and distributed across the culture area surface. In the growth curve, two distinct phases, lag and log phase, were observed at 15 days. Multipotentiality of the cells was evident after induction of osteogenic, chondrogenic, and adipogenic lineages. Regarding the BM-MSCs’ bioelectrical integration with the BCM, BM-MSCs were anchored in the BCM in the first 24 h. On day 7 of culture, the cytoplasm was scattered, and on day 14, the cells were fully integrated with the biomaterial. We also observed significant macrophage activation; analysis of the MTT assay and the concentration of nitric oxide revealed no cytotoxicity of the biomaterial. Conclusion The BCM allowed the expansion and biointegration of bone marrow progenitor cells with a stable cytotoxic profile, thus presenting itself as a biomaterial with potential for tissue engineering

Flavonoides isolados de Sida santaremnensis H. Monteiro (“Guanxuma”) e avaliação das atividades biológicas

Introduction: Sida santaremnensis H. Monteiro (Malvaceae) is a plant popularly known as "vassourinha" or "guanxuma" that has been described as vasorelaxant, antiulcerogenic, antinociceptive and antiedematogenic. Objective: To contribute with the phytochemical and pharmacological profile of Sida santaremnensis through the isolation, purification and determination of chemical constituents of this plant, as well as through the evaluation of the anti-inflammatory and antitumor activity and leishmanicidal effect of the isolated constituent in a greater amount. Methods: The isolation of substances from the plant herein studied was performed with column chromatographic and analytical thin-layer methods and structural determination made by spectroscopic methods, such as Nuclear Magnetic Resonance, Hydrogen and Carbon 13, and comparisons with data from the literature. To assess pharmacological activities, cell viability tests, determination of nitric oxide levels, leishmanicidal activity, among others, were performed. Results: Two flavonoids from S. santaremnensis were obtained, kaempferol (S-1) and kaepferol 3-O-β-D-glycosyl-6’’-α-L-rhamnoside (S-2), the latter in a greater amount. The evaluation of the antitumor activity of the glycosylated flavonoid demonstrated that it does not present hemolytic activity against the human promyelocytic leukemia cell line (HL-60), and that it presented weak leishmanicidal activity. The immunopharmacological evaluation of kaempferol 3-O-β-D-glucosyl-6’’-α-L-rhamnoside revealed that it presents a possible anti-inflammatory action related to the inhibition of the production of nitrite by LPS-stimulated macrophages. Conclusion: These data demonstrate that kaempferol 3-O-β-D-glucosyl-6’’-α-L-rhamnoside has low toxicity, in addition to antileishmania, anti-inflammatory and immunomodulatory properties, which makes it a therapeutic potential for infectious and inflammatory diseases mediated by macrophages.Sida santaremnensis H. Monteiro (Malvaceae) é uma planta conhecida popularmente como "vassourinha" ou "guanxuma", que vem sendo descrita como vasorelaxante, antiulcerogênica, antinociceptiva e antiedematogênica. Objetivou-se contribuir com o estudo fitoquímico e farmacológico de Sida santaremnensis a partir do isolamento e determinação estrutural de constituintes químicos dessa planta, bem como pela avaliação da atividade anti-inflamatória e antitumoral do constituinte isolado em maior quantidade. O isolamento de substâncias da planta estudada foi realizado por métodos cromatográficos em coluna e em camada delgada analítica e a determinação estrutural feita por métodos espectroscópicos, como a Ressonância Magnética Nuclear de Hidrogênio e Carbono 13, e comparações com a literatura. Para avaliação das atividades farmacológicas foram realizados ensaio de viabilidade celular, determinação dos níveis de óxido nítrico, atividade leishmanicida, entre outros. Foram obtidos como resultados dois flavonoides de S. santaremnensis, o canferol (S-1) e canferol 3-O-β-D-glycosyl-6’’-α-L-ramnosídeo (S-2), este último em maior quantidade. A avaliação da atividade antitumoral do flavonoide glicosilado demonstrou que ele não apresenta atividade hemolítica nem citotóxica contra linhagem de células de leucemia promielocítica humana (HL-60), porém apresentou fraca atividade leishmanicida. A avaliação imunofarmacológica dessa mesma substância revelou uma possível ação anti-inflamatória relacionada à inibição da produção de nitrito por macrófagos estimulados com LPS. Conclui-se que estes dados demonstram que o canferol 3-O-β-D-glicosil-6’’-α-L-ramnosídeo apresenta baixa toxicidade, além de propriedades antileishmania, anti-inflamatória e imunomoduladora, o que o torna um potencial terapêutico para doenças infecciosas e inflamatórias mediadas por macrófagos

Influence of Plant Age on Chemical Composition, Antimicrobial Activity and Cytotoxicity of <i>Varronia curassavica</i> Jacq. Essential Oil Produced on an Industrial Scale

Considering the therapeutic potential of Varronia curassavica Jacq. essential oil and the great value in the pharmaceutical market, this study aims to evaluate the influence of plant age on the chemical composition and biological activities of V. curassavica Jacq. essential oil. The plant age is a parameter that can influence the chemical composition of the essential oil, as well as its pharmacological potential. For this purpose, essential oils from aerial parts of V. curassavica produced at different ages (4, 10, 14 and 18 months-age) were used. According to chromatograms obtained by GC-MS, the essential oils were mainly composed of α-pinene, trans-caryophyllene, α-santalene, alloaromadendrene and α-humulene. The chemical composition of V. curassavica essential oils varied qualitatively and quantitatively with the aging of the plants, and the essential oils from plants at 18 month-age appeared to be the most distinct from the others. The tested essential oil samples showed inhibitory activity against Candida albicans (MIC = 1000 µg/mL) but did not show antibacterial activity against the tested bacteria. The cytotoxic activity levels against the murine macrophages varied among the oils extracted from the plants at different ages; the IC50 values of the essential oils increased with age (171.90 µg/mL at 18 month-age). More studies should be carried out to assess whether age also affects the therapeutic effects of essential oils, resulting in the manufacture of plant-derived formulations that balance production costs, toxicity and therapeutic effects

Novel Scaffold Based on Chitosan Hydrogels/Phthalated Cashew Gum for Supporting Human Dental Pulp Stem Cells

Hydrogels are structures that have value for application in the area of tissue engineering because they mimic the extracellular matrix. Naturally obtained polysaccharides, such as chitosan (CH) and cashew gum, are materials with the ability to form polymeric networks due to their physicochemical properties. This research aimed to develop a scaffold based on chitosan and phthalated cashew tree gum and test it as a support for the growth of human mesenchymal stem cells. In this study, phthalation in cashew gum (PCG) was performed by using a solvent-free route. PCG-CH scaffold was developed by polyelectrolyte complexation, and its ability to support adherent stem cell growth was evaluated. The scaffold showed a high swelling rate. The pore sizes of the scaffold were analyzed by scanning electron microscopy. Human dental pulp stem cells (hDPSCs) were isolated, expanded, and characterized for their potential to differentiate into mesenchymal lineages and for their immunophenotypic profile. Isolated mesenchymal stem cells presented fibroblastoid morphology, plastic adhesion capacity, and differentiation in osteogenic, adipogenic, and chondrogenic lineages. Mesenchymal stem cells were cultured in scaffolds to assess cell adhesion and growth. The cells seeded on the scaffold showed typical morphology, attachment, and adequate distribution inside the matrix pores. Thus, cells seeded in the scaffold may improve the osteoinductive and osteoconductive properties of these biomaterials

Solid Lipid Nanoparticles from <i>Platonia insignis</i> Seeds, a Brazilian Amazon Fruit: Characterization, In Vitro and In Vivo Toxicological and Antioxidant Activities

Solid Lipid Nanoparticles (SLNs) are drug delivery systems with important advantages over conventional nanosystems. Considering previously reported pharmacological and physicochemical properties of Platonia insignis seed butter (BBI), this work aimed at developing, characterizing and performing toxicological and antioxidant studies of SLNs produced from BBI. The GC-MS analysis identified palmitic and oleic acids as the major compounds. Three SLN prototypes were developed through high-shear homogenization followed by ultrasonication. During a 180-day stability evaluation, the formulation SLN/TW-1.5 presented greater stability since pH was around 6.0, as well as a lesser variation of the PdI (Polydispersity Index), particle size and Zeta Potential (ZP), confirmed with Raman Spectroscopy and Atomic Force Microscopy (AFM). The CC50 in macrophages was around 249.4 µg∙mL−1 for BBI, whereas the CC50 value for SLN/TW-1.5 was 45.2 µg∙mL−1. Electron Paramagnetic Resonance (EPR) showed a marked in vitro antioxidant activity for BBI and SLN/TW-1.5. After in vivo SLN/TW-1.5 administration in Zophobas morio larvae, assessment of reduced glutathione (GSH), nitrite (NO2−) and myeloperoxidase (MPO) demonstrated antioxidant activity. Thus, the intrinsic physicochemical properties of BBI allowed the development of an optimized nanoformulation with high stability indexes, besides the great potential for antioxidant applications