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1.
Int J Biol Macromol ; 252: 126453, 2023 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-37619683

RESUMO

Serine proteases play crucial biological roles and have their activity controlled by inhibitors, such as the EcTI, a serine protease inhibitor purified from Enterolobium contortisiliquum seeds, which has anticancer activity. This study aimed to conjugate EcTI with quantum dots (QDs), fluorophores with outstanding optical properties, and investigate the interaction of QDs-EcTI nanoprobe with cancer cells. The conjugation was evaluated by fluorescence correlation spectroscopy (FCS) and fluorescence microplate assay (FMA). EcTI inhibitory activity after interaction with QDs was also analyzed. From FCS, the conjugate presented a hydrodynamic diameter about 4× greater than bare QDs, suggesting a successful conjugation. This was supported by FMA, which showed a relative fluorescence intensity of ca. 3815% for the nanosystem, concerning bare QDs or EcTI alone. The EcTI inhibitory activity remained intact after its interaction with QDs. From flow cytometry analyses, approximately 62% of MDA-MB-231 and 90% of HeLa cells were labeled with the QD-EcTI conjugate, suggesting that their membranes have different protease levels to which EcTI exhibits an affinity. Concluding, the QD-EcTI represents a valuable nanotool to study the interaction of this inhibitor with cancer cells using fluorescence-based techniques with the potential to unravel the intricate dynamics of interplays between proteases and inhibitors in cancer biology.


Assuntos
Fabaceae , Neoplasias , Pontos Quânticos , Humanos , Inibidores da Tripsina/farmacologia , Células HeLa , Fabaceae/química , Serina Proteases , Corantes
2.
Parasit Vectors ; 16(1): 273, 2023 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-37559134

RESUMO

BACKGROUND: The resistance of a Culex quinquefasciatus strain to the binary (Bin) larvicidal toxin from Lysinibacillus sphaericus is due to the lack of expression of the toxin's receptors, the membrane-bound Cqm1 α-glucosidases. A previous transcriptomic profile of the resistant larvae showed differentially expressed genes coding Cqm1, lipases, proteases and other genes involved in lipid and carbohydrate metabolism. This study aimed to investigate the metabolic features of Bin-resistant individuals by comparing the activity of some enzymes, energy reserves, fertility and fecundity to a susceptible strain. METHODS: The activity of specific enzymes was recorded in midgut samples from resistant and susceptible larvae. The amount of lipids and reducing sugars was determined for larvae and adults from both strains. Additionally, the fecundity and fertility parameters of these strains under control and stress conditions were examined. RESULTS: Enzyme assays showed that the esterase activities in the midgut of resistant larvae were significantly lower than susceptible ones using acetyl-, butyryl- and heptanoyl-methylumbelliferyl esthers as substrates. The α-glucosidase activity was also reduced in resistant larvae using sucrose and a synthetic substrate. No difference in protease activities as trypsins, chymotrypsins and aminopeptidases was detected between resistant and susceptible larvae. In larval and adult stages, the resistant strain showed an altered profile of energy reserves characterized by significantly reduced levels of lipids and a greater amount of reducing sugars. The fertility and fecundity of females were similar for both strains, indicating that those changes in energy reserves did not affect these reproductive parameters. CONCLUSIONS: Our dataset showed that Bin-resistant insects display differential metabolic features co-selected with the phenotype of resistance that can potentially have effects on mosquito fitness, in particular, due to the reduced lipid accumulation.


Assuntos
Bacillus , Toxinas Bacterianas , Culex , Animais , Feminino , Toxinas Bacterianas/metabolismo , Culex/metabolismo , Lipídeos , Larva/genética
3.
Arch Insect Biochem Physiol ; 113(1): e21872, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-35112391

RESUMO

This study evaluated the effects of acute exposure of Aedes aegypti third instar (L3 ) larvae to the saline extract of Opuntia ficus-indica cladodes on the biological cycle and fertility of the emerging adults. For this, larvae were treated for 24 h with the extract at » LC50 (lethal concentration to kill 50% of larvae), ½ LC50 or LC50 ; the development and reproduction of the emerged adults were evaluated after a recovery period of 9 days. The resistance of proteins in the extract to hydrolysis by L3 digestive enzymes and histomorphological alterations in the larval midgut were also investigated. The extract contained lectin, flavonoids, cinnamic derivatives, terpenes, steroids, and reducing sugars. It showed a LC50 of 3.71% for 48 h. The data indicated mean survival times similar in control and extract treatments. It was observed development delay in extract-treated groups, with a lower number of adults than in control. However, the females that emerged laid similar number of eggs in control and treatments. Histological evaluation revealed absence of bacterial and fungal microorganisms in the food content in midguts from larvae treated with cladode extract. Electrophoresis revealed that three polypeptides in the extract resisted to hydrolysis by L3 digestive proteases for 90 min. The lectin activity was not altered even after 24-h incubation with the enzymes. In conclusion, the extract from O. ficus-indica can delay the development of Ae. aegypti larvae, which may be linked to induction of an axenic environment at larval midgut and permanence of lectin activity even after proteolysis.


Assuntos
Aedes , Inseticidas , Opuntia , Feminino , Animais , Lectinas/química , Larva , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Inseticidas/farmacologia
4.
Parasitol Int ; 92: 102656, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36007704

RESUMO

Allied to the problem of gastrointestinal parasites, especially Haemochus contortus, the use of lectins of plant origin has contributed to the research of alternative anthelmintics. The nematicidal effect of a water soluble Moringa oleifera lectin (WSMoL) was investigated in an experimental model with H. contortus infected Wistar rodents. Three concentrations were tested orally: 5 mg/kg, 2.5 mg/kg and 1 mg/kg. The reduction in the number of larvae recovered in the experimental groups was analyzed, as well as biochemical, hematological and histological parameters. Treatments with 5, 2.5 and 1 mg/kg of WSMoL reduced the number of larvae recovered of animals by 74.7 %, 72.8 % and 66 %, respectively. Untreated infected animals had anemia, moderate mononuclear multifocal hepatitis, vascular congestion in the liver and kidneys, white pulp hyperplasia in the spleen, and presence of eosinophils in the intestine. Infected animals treated with 5 mg/kg of WSMoL showed liver with moderate bleeding, kidney with vascular congestion, spleen with white pulp hyperplasia and intestine with moderate presence of mononuclear cells. An increase in the serum level of glutamic pyruvic transaminase and a reduction in the level of hemoglobin (p < 0.001) were also observed in this group when compared to the uninfected group. However, the administered concentrations of 2.5 and 1 mg/kg of WSMoL were both satisfactory in terms of reducing the number of recovered larvae and not promoting negative changes in the biochemical, hematological and histological parameters evaluated. These results indicate an in vivo nematicidal effect of WSMoL on the H. contortus parasite.


Assuntos
Anti-Helmínticos , Haemonchus , Moringa oleifera , Animais , Lectinas , Roedores , Água , Hiperplasia , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Antinematódeos , Larva
5.
An Acad Bras Cienc ; 94(3): e20211297, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35920491

RESUMO

Angiogenesis (budding of new blood vessels) is involved in several processes, including the development of embryos and growth of tumors. Schinus terebinthifolia leaves express an antitumor lectin (SteLL). This work hypothesized that SteLL can interfere with the formation of a vascular network from preexisting vessels. To test this hypothesis, the effect of SteLL on the angiogenesis process was assessed using an in vivo model of yolk sac membrane of Coturnix japonica embryos. SteLL was isolated with purification factor of 46.6. As expected, polyacrylamide gel electrophoresis (PAGE) for native basic proteins confirmed the homogeneity and PAGE in presence of dodecyl sodium sulphate revealed a single 14-kDa polypeptide band. The fractal analysis by box counting and information dimension measurements indicated that SteLL at 1.35 mg/mL significantly decreased by ca. 12% the angiogenesis within the C. japonica yolk sac membrane regarding the control. The inhibition of the vascular network formation in the yolk sac membrane resulted in decreased blood supply to the embryos. Consequently, the area of embryos was significantly reduced by 9.2% regarding the control, which corroborated with the antiangiogenic activity of SteLL. The findings implicate SteLL as an antiangiogenic agent and add to the panel of biological activities of this lectin.


Assuntos
Anacardiaceae , Coturnix , Inibidores da Angiogênese/farmacologia , Animais , Lectinas/farmacologia , Folhas de Planta
6.
Sci Total Environ ; 841: 156795, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-35732235

RESUMO

Flocculation has been proved an efficient method for microalgal biomass harvesting, but some coagulant agents may have adverse effects on microalgae growth, making the reuse of the medium unfeasible. In this study, Haematococcus pluvialis was harvested by different flocculants, and the feasibility of the reuse of the culture medium was evaluated. Results suggested that both inorganics, polyaluminum chloride (PA) and ferric chloride (FC), and organics, extracted from Moringa oleifera seed (MSE) and chitosan (CH) resulted in efficient flocculation - flocculation efficiency above 99 %. However, using PA and FC had adverse effects on the astaxanthin recovery from haematocysts - losses of 58.6 and 73.5 %, respectively. Bioflocculants in the reused medium also had higher growth performance than inorganic ones. Furthermore, bioflocculants in reused medium increase the contents of ß-carotene, astaxanthin, and linolenic acid. This investigation demonstrated that using MSE and CHI for harvesting H. pluvialis enables the water reusability from a flocculated medium.


Assuntos
Clorofíceas , Microalgas , Biomassa , Floculação , Água , Xantofilas
7.
Biomass Convers Biorefin ; : 1-9, 2022 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-35582461

RESUMO

Bio-flocculation is a sustainable low-cost harvesting technique for microalgae biomass production; however, it is generally less efficient than chemical flocculants. This study aims to investigate the efficiency of Moringa oleifera seeds as a bio-flocculant for harvesting Tetradesmus dimorphus biomass. Four extracts from integral and residual (seeds without lipids) biomass of M. oleifera seeds using salt or aqueous solutions were used at four concentrations (100, 200, 300, and 400 ppm). Flocculation efficiency (FE) increased as the pH decreased. The addition of the extracts reduced the pH of the cultures, dispensing pH adjustment after dosing the flocculating agent. Salt extracts exhibited higher flocculation efficiency than aqueous extracts. The highest flocculation efficiency (~ 98%) was obtained using a salt extract of residual biomass of seeds in any concentration varying from 100 to 400 ppm. The predicted values obtained from a data modeling using response surface methodology approached the real values (r 2 = 0.9382), resulting in an adequate optimization of the flocculant concentration of ~ 335 ppm and pH 5.6 for a predicted FE of ~ 106%. The findings of the present study confirmed that the salt extract from residual biomass of M. oleifera seeds is an effective bio-flocculant for T. dimorphus biomass harvesting.

8.
Exp Parasitol ; 218: 108004, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32961172

RESUMO

Haemonchus contortus is a hematophagous parasite causing damage to the production of ruminant animals throughout the world. This study evaluated the in vitro effect of proteins from Moringa oleifera (WSMoL - Water Soluble M. oleifera Lectin and cMoL - coagulant M. oleifera Lectin) on the motility of infective larvae and adult male and female worms of H. contortus. The specific activity of total proteases and the morphology of the worms exposed to the lectins were observed. Both lectins inhibited motility of all parasite stages tested. WSMoL and cMoL at 500 µg mL-1 interfered in the motility of larvae. Values of 11.1% and 8.1% were the lowest motility indices of larvae with sheath, and 30.6% and 16.4% were the lowest motility indices of exsheathed larvae treated with WSMoL and cMoL, respectively. In 1 mg mL-1 solutions of WSMoL and of cMoL, the motility index of adult male worms was 23.3% (p < 0.001) and 20% (p < 0.001), while the motility index of adult female worms was 63.3% (p > 0.05) and 26.6% (p < 0.001), respectively. Greater proteolytic activity was detected in extracts obtained from adult worms, male and female, after incubation with the lectins. Morphological changes caused by the lectins were revealed by changes in the crests of the cuticle, in the longitudinal striations and at the vulva.


Assuntos
Haemonchus/efeitos dos fármacos , Moringa oleifera/química , Lectinas de Plantas/farmacologia , Sementes/química , Animais , Feminino , Haemonchus/enzimologia , Haemonchus/fisiologia , Haemonchus/ultraestrutura , Larva/efeitos dos fármacos , Masculino , Microscopia Eletrônica de Varredura , Movimento/efeitos dos fármacos , Peptídeo Hidrolases/metabolismo , Lectinas de Plantas/administração & dosagem , Lectinas de Plantas/isolamento & purificação
9.
Methods Appl Fluoresc ; 8(3): 035009, 2020 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-32633256

RESUMO

Carbohydrates perform important physiological functions in eukaryotic and prokaryotic cells. Indeed, alterations in glycan patterns may be associated with disorders. The analysis of these sugars can be reached using nanoprobes composed by lectins associated with fluorescent nanoparticles. This study reports the conjugation of a galactose-binding lectin (BmoLL) isolated from Bauhinia monandra leaves with quantum dots (QDs) by adsorption. QDs-BmoLL conjugates showed bright fluorescence and the hemagglutination assay revealed that the lectin preserved its carbohydrate-binding ability after the conjugation. To evaluate the efficiency/specificity of the bioconjugate, ABO human red blood cells (RBCs) were used as biological models and the labeling was analyzed by flow cytometry. Among ABO blood groups, higher labeling (71.7 ± 5.9%) was detected for B-type RBCs, whose antigens have galactose in their structure. The specificity of labeling was confirmed since A- and O-types RBCs incubated with QDs-BmoLL, as well as B-type cells incubated with previously galactose-inhibited conjugates, were labeled below 6%. In AB-type RBCs, which simultaneously have B and A (N-acetylgalactosamine) antigens on their membrane, the labeling was ca. 14.1 ± 4.8%. Therefore, a successful conjugation was reached and QDs-BmoLL conjugates can be considered promising fluorescent nanoprobes for biological investigations.


Assuntos
Bauhinia/química , Eritrócitos/química , Nanopartículas/química , Folhas de Planta/química , Pontos Quânticos/química , Humanos
10.
Microbiol Res ; 230: 126330, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31541842

RESUMO

Glycoconjugates found on cell walls of Candida species are fundamental for their pathogenicity. Laborious techniques have been employed to investigate the sugar composition of these microorganisms. Herein, we prepared a nanotool, based on the fluorescence of quantum dots (QDs) combined with the specificity of Cramoll lectin, to evaluate glucose/mannose profiles on three Candida species. The QDs-Cramoll conjugates presented specificity and bright fluorescence emission. The lectin preserved its biological activity after the conjugation process mediated by adsorption interactions. The labeling of Candida species was analyzed by fluorescence microscopy and quantified by flow cytometry. Morphological analyses of yeasts labeled with QDs-Cramoll conjugates indicated that C. glabrata (2.7 µm) was smaller when compared to C. albicans (4.0 µm) and C. parapsilosis sensu stricto (3.8 µm). Also, C. parapsilosis population was heterogeneous, presenting rod-shaped blastoconidia. More than 90% of cells of the three species were labeled by conjugates. Inhibition and saturation assays indicated that C. parapsilosis had a higher content of exposed glucose/mannose than the other two species. Therefore, QDs-Cramoll conjugates demonstrated to be effective fluorescent nanoprobes for evaluation of glucose/mannose constitution on the cell walls of fungal species frequently involved in candidiasis.


Assuntos
Candida/química , Corantes Fluorescentes/química , Glucose/análise , Lectinas/química , Manose/análise , Microscopia de Fluorescência/métodos , Candida/crescimento & desenvolvimento , Candida/isolamento & purificação , Candida/metabolismo , Candidíase/diagnóstico , Candidíase/microbiologia , Parede Celular/química , Parede Celular/metabolismo , Glucose/metabolismo , Humanos , Manose/metabolismo , Microscopia de Fluorescência/instrumentação , Nanopartículas/química , Pontos Quânticos/química
11.
Pest Manag Sci ; 76(2): 730-736, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31386279

RESUMO

BACKGROUND: Lectins from Moringa oleifera seeds (WSMoL), Myracrodruon urundeuva bark (MuBL), and heartwood (MuHL) are larvicidal agents against Aedes aegypti; in addition, WSMoL is an ovicidal agent against this mosquito. In this work, we evaluated the ovicidal activity of MuBL and MuHL by determining the concentrations that reduce the hatching rates by 50% in 72 h (EC50 ). The effects of WSMoL, MuBL, and MuHL on the ultrastructure of the eggs' surface were assessed by scanning electron microscopy (SEM). In addition, the ability of these lectins to penetrate the eggs was investigated by using conjugates of the lectins with fluorescein isothiocyanate (FITC). RESULTS: MuBL and MuHL were ovicidal agents with EC50 of 0.26 and 0.80 mg/mL (260 and 800 ppm), respectively. SEM images of eggs treated with WSMoL for 24 h revealed discontinuity of the exochorionic network and the absence of the exochorionic cells and their tubercles. After 48 and 72 h of incubation, strong deformation and degeneration of egg surfaces were observed. In MuBL and MuHL-treated eggs, the presence of lumps on the surface of the eggs, disappearance of the exochorionic network and the decrease and deformation of tubercles were observed. Lastly, fluorescence microscopy revealed that the three lectins were able to enter the eggs and reach the digestive tract of the embryos. CONCLUSION: WSMoL, MuBL, and MuHL are ovicidal agents on A. aegypti that have differing efficiencies in terms of how they cause alterations in the chorionic surface and in terms of their ability to penetrate the eggs. © 2019 Society of Chemical Industry.


Assuntos
Aedes , Anacardiaceae , Moringa oleifera , Animais , Inseticidas , Larva , Lectinas , Óvulo , Extratos Vegetais , Sementes
12.
Colloids Surf B Biointerfaces ; 187: 110672, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31796242

RESUMO

Mosquito-borne arboviruses compromise human health worldwide. Due to resistance to chemical insecticides, natural compounds have been studied to combat mosquitoes. Previous works have demonstrated a larvicidal activity of the water-soluble Moringa oleifera lectin (WSMoL) against Aedes aegypti, suggesting a mechanism of action based on the interaction between lectin and chitin present in the larvae's peritrophic matrix. In this work, it was investigated the WSMoL activity against Aedes aegypti larvae, by using luminescent bioconjugates of WSMoL conjugated to l-glutathione capped CdTe quantum dots. The conjugation was confirmed by ITC experiments, presenting high enthalpy associated to hydrogen bond interactions between nanoparticles and lectins. The bioconjugate luminescence stability was evaluated by the quantum yield (QY) at different pHs, ionic strengths and heat treatment time. The best parameters reached were pH 7.0, absence of electrolytes and heat treatment, giving QY = 4.4 %. The larvae were exposed to the bioconjugates and analyzed by confocal and fluorescence microscopy. CdTe-WSMol were detected along the entire midgut tract, suggesting a strong interaction with peritrophic matrix and lumen of the Aedes aegypti.


Assuntos
Compostos de Cádmio/química , Glutationa/química , Larva/efeitos dos fármacos , Lectinas/química , Moringa oleifera/química , Pontos Quânticos/química , Telúrio/química , Aedes/efeitos dos fármacos , Animais , Biomarcadores/análise , Temperatura Alta , Ligação de Hidrogênio , Concentração de Íons de Hidrogênio , Luminescência , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Pontos Quânticos/ultraestrutura , Difração de Raios X
13.
Microbiologyopen ; 8(11): e869, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31099495

RESUMO

This work aimed to evaluate the effects of CasuL on growth and viability of 15 mastitis isolates from cows and goats, to determine the synergistic potential between CasuL and antibiotics, and to investigate the effects on bacterial ultrastructure and antibiofilm activity. The lectin inhibited the growth of Staphylococcus isolates from either bovine (Ssp6PD and Sa) or caprine (Ssp5D and Ssp01) mastitis. The minimal inhibitory concentrations were ranged from 3.75 to 15 µg/ml. Synergistic effect was observed for CasuL-tetracycline against Sa and Ssp6PD and CasuL-ampicillin against Ssp01. No structural damage was observed under the scanning electron microscope in CasuL treatments. Flow cytometry analysis using thiazol orange and propidium iodide demonstrated that CasuL was unable to reduce the cell viability of the isolates tested. At sub-inhibitory concentrations, CasuL reduced biofilm formation by the isolates Sa and Ssp5D. However, CasuL-tetracycline and CasuL-ampicillin combinations inhibited biofilm formation by Ssp6PD and Ssp01, respectively. In conclusion, CasuL is a bacteriostatic and antibiofilm agent against some mastitis isolates and displayed a synergistic potential when used in combination with either ampicillin (against one isolate) or tetracycline (against two isolates). The results stimulate the evaluation of CasuL for the treatment of mastitis, particularly when used in conjunction with antibiotics.


Assuntos
Antibacterianos/farmacologia , Doenças dos Bovinos/microbiologia , Fabaceae/química , Doenças das Cabras/microbiologia , Lectinas/farmacologia , Mastite/veterinária , Staphylococcus/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Bovinos , Sinergismo Farmacológico , Cabras , Lectinas/isolamento & purificação , Mastite/microbiologia , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus/crescimento & desenvolvimento , Staphylococcus/isolamento & purificação
14.
Pest Manag Sci ; 75(5): 1212-1222, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30306668

RESUMO

The midgut of insects is involved in digestion, osmoregulation and immunity. Although several defensive strategies are present in this organ, its organization and function may be disturbed by some insecticidal agents, including bioactive proteins like lectins and protease inhibitors (PIs) from plants. PIs interfere with digestion, leading to poor nutrient absorption and decreasing amino acid bioavailability. Intake of PIs can delay development, cause deformities and reduce fertility. Ingestion of PIs may lead to changes in the set of proteases secreted in the insect gut, but this response is often insufficient and results in aggravation of the malnutrition status. Lectins are proteins that are able to interact with glycoconjugates, including those linked to cell surfaces. Their effects on the midgut include disruption of the peritrophic matrix, brush border and secretory cell layer; induction of apoptosis and oxidative stress; interference with nutrient absorption and transport proteins; and damaging effects on symbionts. In addition, lectins can cross the intestinal barrier and reach the hemolymph. The establishment of resistant insect populations due to selective pressure resulting from massive use of a bioactive protein is an actual possibility, but this can be minimized by the multiple mode-of-action of these proteins, mainly the lectins. © 2018 Society of Chemical Industry.


Assuntos
Sistema Digestório/efeitos dos fármacos , Controle de Insetos/métodos , Insetos/anatomia & histologia , Lectinas/farmacologia , Plantas/química , Inibidores de Proteases/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Animais , Insetos/fisiologia
15.
Protein Pept Lett ; 25(3): 295-301, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29384049

RESUMO

BACKGROUND: Moringa oleifera is used in traditional medicine as well as in food, cosmetic, and pharmaceutical industries. Water-soluble M. oleifera lectin (WSMoL) is an anionic protein isolated from the seeds of this tree. Until now, immune responses promoted by this lectin in human PBMC have not been investigated. OBJECTIVE: The main objective of this study was to investigate the immunomodulatory effects of WSMoL on human PBMC through measurement of lymphocytes subsets, cytokine and nitric oxide levels. METHODS: Human peripheral blood mononuclear cells (PBMC) were isolated through Ficoll technique, were incubated with WSMoL (10 µg/mL) for 24, 48 and 72 hours, and was performed immunophenotyping assay of lymphocytes and monocytes. Culture supernatants were used to determined cytokine and nitric oxide levels. Assays with cells subsets and cytokine production were performed through cytometry. Nitric oxide release assay was determinate by spectrophotometry. RESULTS: WSMoL induced the release of the cytokines TNF-α, IL-2, IL-6, IL-10 as well as nitric oxide. Incubation of PBMC with this lectin also led to activation of CD8+ T lymphocytes. CONCLUSION: WSMoL promotes immunomodulation in human PBMC inducing a potential wound healing profile and, in future in vivo assays, can be evaluated as adjuvant in immunosuppressive diseases and wound repair.


Assuntos
Fatores Imunológicos/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Moringa oleifera/química , Lectinas de Plantas/farmacologia , Sementes/química , Adulto , Sobrevivência Celular , Citocinas/metabolismo , Humanos , Fatores Imunológicos/isolamento & purificação , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/imunologia , Óxido Nítrico/metabolismo , Extratos Vegetais/isolamento & purificação , Lectinas de Plantas/isolamento & purificação , Solubilidade , Água , Adulto Jovem
16.
Pest Manag Sci ; 74(7): 1593-1599, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29297969

RESUMO

BACKGROUND: Lectins, carbohydrate-binding proteins, from the bark (MuBL) and leaf (MuLL) of Myracrodruon urundeuva are termiticidal agents against Nasutitermes corniger workers and have been shown to induce oxidative stress and cell death in the midgut of these insects. In this study, we investigated the binding targets of MuBL and MuLL in the gut of N. corniger workers by determining the effects of these lectins on the activity of digestive enzymes. In addition, we used mass spectrometry to identify peptides from gut proteins that adsorbed to MuBL-Sepharose and MuLL-Sepharose columns. RESULTS: Exoglucanase activity was neutralized in the presence of MuBL and stimulated by MuLL. α-l-Arabinofuranosidase activity was not affected by MuBL but was inhibited by MuLL. Both lectins stimulated α-amylase activity and inhibited protease and trypsin-like activities. Peptides with homology to apolipophorin, trypsin-like enzyme, and ABC transporter substrate-binding protein were detected from proteins that adsorbed to MuBL-Sepharose, while peptides from proteins that bound to MuLL-Sepharose shared homology with apolipophorin. CONCLUSION: This study revealed that digestive enzymes and transport proteins found in worker guts can be recognized by MuBL and MuLL. Thus, the mechanism of their termiticidal activity may involve changes in the digestion and absorption of nutrients. © 2018 Society of Chemical Industry.


Assuntos
Anacardiaceae/química , Inseticidas/metabolismo , Isópteros/efeitos dos fármacos , Lectinas de Plantas/metabolismo , Animais , Sistema Digestório/efeitos dos fármacos , Sistema Digestório/enzimologia , Isópteros/enzimologia , Casca de Planta/química , Folhas de Planta/química , Lectinas de Plantas/administração & dosagem
17.
Nat Prod Res ; 32(24): 2940-2944, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29047320

RESUMO

In this study, Moringa oleifera flower extract and a trypsin inhibitor (MoFTI) isolated from it were evaluated for anti-protozoal activity against Trypanosoma cruzi and cytotoxicity to mammalian cells. The presence of flavonoids was remarkable in the HPLC fingerprints of the extract at 254 and 360 nm. Amino acid sequences of peptides derived from in-gel digestion of MoFTI were determined. Both the extract and MoFTI caused lysis of T. cruzi trypomastigotes with LC50/24 h of 54.18 ± 6.62 and 41.20 ± 4.28 µg/mL, respectively. High selectivity indices (7.9 to >12) for T. cruzi cells over murine peritoneal macrophages and Vero cells were found for the extract and MoFTI. The results show that MoFTI is a trypanocidal principle of the flower extract.


Assuntos
Flavonoides , Flores/química , Moringa oleifera/química , Extratos Vegetais/química , Tripanossomicidas , Trypanosoma cruzi/efeitos dos fármacos , Inibidores da Tripsina/isolamento & purificação , Animais , Linhagem Celular , Chlorocebus aethiops , Flavonoides/análise , Macrófagos Peritoneais/efeitos dos fármacos , Mamíferos , Camundongos , Extratos Vegetais/farmacologia , Tripanossomicidas/farmacologia , Inibidores da Tripsina/farmacologia , Células Vero/efeitos dos fármacos
18.
Pest Manag Sci ; 73(5): 991-998, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27530272

RESUMO

BACKGROUND: Myracrodruon urundeuva is a hardwood tree, and its bark, heartwood and leaf contain lectins (MuBL, MuHL and MuLL respectively) with termiticidal activity against Nasutitermes corniger. In this work, the effects of these lectins on the midgut of N. corniger workers were evaluated. RESULTS: The insects were supplied with an artificial diet containing the lectins at their respective LC50 (previously determined). At 48 h after treatment, the midguts were dissected and fixed for histopathology analyses. Toluidine-blue-stained midguts from lectin-treated workers showed disorganisation, with the presence of debris in the lumen and the absence of brush border. Fluorescence microscopy revealed that the numbers of digestive and proliferating cells were lower in lectin-treated individuals than in the control, and caspase-3 staining confirmed the occurrence of cell apoptosis. Enteroendocrine cells were not seen in the treated individuals. The midguts from treated insects showed greater staining for peroxidase than the control, suggesting that the lectins caused oxidative stress. Staining with wheat germ agglutinin conjugated to FITC revealed that the lectins interfered with the integrity of the peritrophic matrix. CONCLUSION: This study showed that termiticidal lectins from M. urundeuva cause severe injuries, oxidative stress and cell death in the midgut of N. corniger workers. © 2016 Society of Chemical Industry.


Assuntos
Anacardiaceae/química , Ingestão de Alimentos , Inseticidas/farmacologia , Mucosa Intestinal/efeitos dos fármacos , Isópteros/efeitos dos fármacos , Lectinas de Plantas/farmacologia , Animais , Suplementos Nutricionais , Isópteros/fisiologia
19.
Front Microbiol ; 7: 1466, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27729901

RESUMO

In the face of increasing bacterial resistance to antibiotics currently in use, the search for new antimicrobial agents has received a boost in recent years, with natural products playing an important role in this field. In fact, several methods have been proposed to investigate the antibacterial activities of natural products. However, given that the ultimate aim is future therapeutic use as novel drugs, it is extremely necessary to elucidate their modes of action, stating the molecular effects in detail, and identifying their targets in the bacterial cell. This review analyzes the application of "omics technologies" to understand the antibacterial mechanisms of bioactive natural products, to stimulate research interest in this area and promote scientific collaborations. Some studies have been specifically highlighted herein by examining their procedures and results (targeted proteins and metabolic pathways). These approaches have the potential to provide new insights into our comprehension of antimicrobial resistance/susceptibility, creating new perspectives for the struggle against bacteria, and leading to the development of novel products in the future.

20.
Protein Pept Lett ; 23(7): 645-9, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27184499

RESUMO

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.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Metais/isolamento & purificação , Moringa oleifera/química , Lectinas de Plantas/química , Lectinas de Plantas/farmacologia , Poluentes Químicos da Água/isolamento & purificação , Escherichia coli/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Hemaglutinação/efeitos dos fármacos , Humanos , Salmonella typhi/efeitos dos fármacos , Sementes/química , Febre Tifoide/tratamento farmacológico , Purificação da Água
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