Maiara Celine de Moura

Evaluation of cytotoxic and anti-inflammatory activities of extracts and lectins from Moringa oleifera seeds.

Background The extract from Moringa oleifera seeds is used worldwide, especially in rural areas of developing countries, to treat drinking water. M. oleifera seeds contain the lectins cmol and WSMoL, which are carbohydrate-binding proteins that are able to reduce water turbidity because of their coagulant activity. Studies investigating the ability of natural products to damage normal cells are essential for the safe use of these substances. This study evaluated the cytotoxic and anti-inflammatory properties of the aqueous seed extract, the extract used by population to treat water (named diluted seed extract in this work), and the isolated lectins cmol and WSMoL. Methodology/Principal Findings The data showed that the aqueous seed extract and cmol were potentially cytotoxic to human peripheral blood mononuclear cells, while WSMoL and diluted seed extract were not cytotoxic. The M. oleifera aqueous seed extract and the lectins cmol and WSMoL were weakly/moderately cytotoxic to the NCI-H292, HT-29 and HEp-2 cancer cell lines and were not hemolytic to murine erythrocytes. Evaluation of acute toxicity in mice revealed that the aqueous seed extract (2.000 mg/kg) did not cause systemic toxicity. The aqueous seed extract, cmol and WSMoL (6.25 µg/mL) and diluted seed extract at 50 µg/mL exhibited anti-inflammatory activity on lipopolyssaccharide-stimulated murine macrophages by regulating the production of nitric oxide, TNF-α and IL-1β. The aqueous seed extract reduced leukocyte migration in a mouse model of carrageenan-induced pleurisy; the myeloperoxidase activity and nitric oxide, TNF-α and IL-1β levels were similarly reduced. Histological analysis of the lungs showed that the extract reduced the number of leukocytes. Conclusion/Significance This study shows that the extract prepared according to folk use and WSMoL may be non-toxic to mammalian cells; however, the aqueous seed extract and cmol may be cytotoxic to immune cells which may explain the immunosuppressive potential of the extract.

Water‐soluble Moringa oleifera lectin interferes with growth, survival and cell permeability of corrosive and pathogenic bacteria

This work evaluated the antibacterial activity of a water‐soluble Moringa oleifera seed lectin (WSMoL) by evaluating its effect on growth, survival and cell permeability of Bacillus sp., Bacillus cereus, Bacillus pumillus, Bacillus megaterium, Micrococcus sp., Pseudomonas sp., Pseudomonas fluorescens, Pseudomonas stutzeri and Serratia marcescens. In addition, the effect of lectin on membrane integrity of most sensitive species was also evaluated. All the tested bacteria are able to cause biocorrosion and some are also responsible for human infections.

CasuL: A new lectin isolated from Calliandra surinamensis leaf pinnulae with cytotoxicity to cancer cells, antimicrobial activity and antibiofilm effect.

This work describes the isolation of a lectin (CasuL) from the leaf pinnulae of Calliandra surinamensis and the evaluation of its cytotoxic, antimicrobial and antibiofilm properties. Proteins from pinnulae extract were precipitated with ammonium sulphate (60% saturation) and submitted to Sephadex G-75 chromatography, which yielded isolated CasuL (purification factor: 113). Native CasuL is an acidic protein (pI 5.82) with a relative molecular mass of 48kDa. This lectin is also an oligomeric protein composed of three subunits and mass spectrometry revealed similarities with a Sorghum bicolor protein. CasuL did not undergo unfolding when heated but changes in conformation and hemagglutinating activity were detected at basic pH. CasuL did not reduce the viability of human peripheral blood mononuclear cells but was toxic to leukemic K562 cells (IC50 67.04±5.78μg/mL) and breast cancer T47D cells (IC50: 58.75±2.5μg/mL). CasuL (6.25-800μg/mL) only showed bacteriostatic effect but was able to reduce biofilm formation by Staphylococcus saprophyticcus and Staphylococcus aureus (non-resistant and oxacillin-resistant isolates). CasuL showed antifungal activity against Candida krusei causing alterations in cell morphology and damage to cell wall. In conclusion, the pinnulae of C. surinamensis leaves contain a thermo-stable lectin with biotechnological potential as cytotoxic, antibiofilm, and antifungal agent.

PgTeL, the lectin found in Punica granatum juice, is an antifungal agent against Candida albicans and Candida krusei

The pomegranate (Punica granatum) sarcotesta contains a chitin-binding lectin (PgTeL) with antibacterial activity against human pathogenic species. In this work, the structural stability of PgTeL was evaluated by fluorimetric analysis and the lectin was evaluated for cytotoxicity to human peripheral blood mononuclear cells (PBMCs) and antifungal activity against Candida albicans and Candida krusei. PgTeL folding was impaired when lectin was incubated at pH≥6.0. On the other hand, the lectin did not undergo unfolding even when heated at 100°C. PgTeL (1, 10, and 100μg/mL) was not cytotoxic to PBMCs. Antifungal activity was detected for C. albicans (MIC: 25μg/mL; MFC: 50μg/mL) and C. krusei (MIC and MFC of 12.5μg/mL). Treatment of yeast cells with PgTeL resulted in decrease of intracellular ATP content even at sub-inhibitory concentrations (½MIC and ¼MIC) and induced lipid peroxidation. In addition, PgTeL damaged the integrity of fungal cell wall of both species, with more pronounced effects in C. krusei. The lectin showed significant antibiofilm activity on C. albicans at sub-inhibitory concentrations (0.195 and 0.39μg/mL). In conclusion, PgTeL is an anti-Candida agent whose action mechanism involves oxidative stress, energetic collapse, damage to the cell wall and rupture of yeast cells.

Multi-effect of the water-soluble Moringa oleifera lectin against Serratia marcescens and Bacillus sp.: antibacterial, antibiofilm and anti-adhesive properties.

To evaluate the antibiofilm potential of water‐soluble Moringa oleifera seed lectin (WSMoL) on Serratia marcescens and Bacillus sp.

Evaluation of Moringa oleifera Seed Lectin as a Metal Remover in Aqueous Solutions.

The efficacy of Moringa oleifera seed lectin (WSMoL) as a metal remover in water and the effects of metals on its hemagglutinating and antibacterial activities were determined. Aqueous metal solutions were incubated with WSMoL for 8 h at 4°C and the concentrations of metals were determined using atomic absorption spectroscopy. Hemagglutination and antibacterial assays were conducted with WSMoL and lectin exposed or not to the metals. The removal efficiency of WSMoL was 49.00%, 53.21%, 71.45%, 55.42%, 69.88%, 62.14%, and 49.36% for Cd+2, Pb+2, Cu+2, Zn+2, Mg+2, Mn+2, and Al+3, respectively. WSMoL showed bacteriostatic and bactericidal activities against Escherichia coli and Salmonella enterica serovar Enteritidis. However, hemagglutinating and antibacterial activities were impaired after exposure to metals. In conclusion, WSMoL efficiently removed metals present in water but the interaction with metals impaired lectin carbohydrate-binding ability and antibacterial activity. This should be considered when properties of WSMoL other than metal removal are desired.

Application of the advanced oxidative process on the degradation of the green leaf and purple açaí food dyes with kinetic monitoring and artificial neural network modelling

The study evaluated the advanced oxidative processes concerning the degradation of green leaf and purple açaí dyes, as well as the prediction of data through artificial neural networks (ANNs). It was verified that percentage of degradation on the wavelengths (λ) of 215, 248, 523 and 627 nm was 5.95, 49.99, 98.17 and 95.99%, respectively, when UV/H2O2 action and UV-C radiation was applied. A non-linear kinetic model proposed by Chan and Chu presented a good fit to the experimental data, reaching an R2 value between 0.978 and 0.999, for the studied λ. Within the ANN simulations through Statistica 6.0, the multilayer perceptron (MLP) (3-9-4) presented a better fit to the experimental data. However, higher values of R² were obtained when utilizing the sklearn package with Python language and an MLP (4-5-4) model. Assays with Staphylococcus aureus and Staphylococcus pyogenes bacteria isolates were performed and it was verified that after employing the UV/H2O2 process, there was a decrease in the toxicity of the solution of dyes. In evaluating S. aureus toxicity, normal growth was observed. However, for S. pyogenes bacteria, it was found that when using the UV/H2O2 process, toxicity was evidenced at post-treatment solution concentrations of 100, 70 and 50%.

Degradation of textile dyes Remazol Yellow Gold and reactive Turquoise: optimization, toxicity and modeling by artificial neural netwoks

In this work, the degradation of Remazol Yellow Gold RNL-150% and Reactive Turquoise Q-G125 were investigated using AOP: photolysis, UV/H2O2, Fenton and photo-Fenton. It was found that the photo-Fenton process employing sunlight radiation was the most efficient, obtaining percentages of degradation above 87%. The ideal conditions for the degradation of the dyes were determined from a factorial design 23 and study of the [H2O2] ([H2O2] equal to 100 mg·L-1); [Fe] equal to 1 mg·L-1 and pH between 3 and 4. In the kinetic study, a degradation of more than 97% was obtained after 150 min for the chromophoric groups and 91% for the aromatic compounds. The experimental data obtained presented a good fit to the nonlinear kinetic model. The model of artificial neural networks multilayer perceptron (MLP) (4-11-5) using the software Statistica 8.0 enabled the modeling of the degradation process and showed a better prediction of the data. The toxicity to the seeds of Lactuca sativa and the bacteria Escherichia coli and Salmonella enteritidis allowed to evaluate the effectiveness of the process. The results of this study suggest that the use of photo-Fenton process with sunlight radiation is an effective way to degrade the dyes under study.

Antimicrobial potential of Alpinia purpurata lectin (ApuL): Growth inhibitory action, synergistic effects in combination with antibiotics, and antibiofilm activity

The Alpinia purpurata inflorescence contains a lectin (ApuL), which has immunomodulatory activities on human cells. In the present work, it was evaluated the antibacterial and antifungal effects of ApuL against human pathogens. ApuL showed bacteriostatic activity against non-resistant (UFPEDA-02) and an oxacillin-resistant isolate (UFPEDA-672) of Staphylococcus aureus with minimal inhibitory concentrations (MIC50) of 50 and 400 μg/mL, respectively. In addition, it showed bactericidal effect on the non-resistant isolate (minimal bactericidal concentration: 200 μg/mL). For Candida albicans and Candida parapsilosis, ApuL showed fungistatic effect (MIC50: 200 and 400 μg/mL, respectively). The lectin was able to impair the viability of the microorganism cells, as indicated by propidium iodide (PI) staining. Analysis of growth curves, protein leakage, and ultrastructural changes supported that ApuL acts through distinct mechanisms on S. aureus isolates. Ultrastructural analysis of ApuL-treated Candida cells revealed malformations with elongations and bulges. ApuL-oxacillin combination showed synergistic effect on the oxacillin-resistant isolates UFPEDA-670 and 671, which were not sensitive to lectin alone. Synergism was also detected for ApuL-ceftazidime against a multidrug-resistant isolate of Pseudomonas aeruginosa. Synergistic action of ApuL-fluconazole was detected for C. parapsilosis, which was insensitive to the drug alone. Biofilm formation by S. aureus non-resistant isolate and C. albicans was remarkably inhibited by ApuL at sub-inhibitory concentrations. In conclusion, ApuL showed differential effects on non-resistant and resistant bacterial isolates, was active against Candida species, and showed synergistic action in combination with antibiotics.

NEMATICIDAL ACTIVITY OF A WATER SOLUBLE LECTIN FROM SEEDS OF Moringa oleifera

Gastrointestinal nematodes are the major cause of animal diseases, and the indiscriminate use of synthetic anthelmintic contributes to the development of drug resistance. Natural bioproducts are noteworthy since they have proved to be promising for alternative antiparasitic therapies. This study evaluated the in vitro effect of WSMoL (water soluble Moringa oleifera lectin) on hatching of eggs and on the development of early-stage larvae of gastrointestinal nematodes from naturally infected goats. In addition, the interference of WSMoL on activity of proteases was determined and the affinity of the lectin for glycosylated molecules of these parasites was investigated using fluorescein isothiocyanate (FITC)-labeled WSMoL. WSMoL at 250μgmL-1 interfered on hatching of eggs (40.4% of not hatched eggs; p=0.018), and on larval development (stages L1-L3) (IC50 of 78.22μgmL-1). The activity of secreted proteases showed a significant increase in the presence of WSMoL (307.14U/mg-1031U/mg). FITC-labeled WSMoL recognized embryonic egg content and larval content after hatching, which suggests that WSMoL interact with intestinal glycoconjugate receptors in the embryo, as well as in cuticle of the larvae.