RESUMO
This study characterized the binding mechanisms of the lectin cMoL (from Moringa oleifera seeds) to carbohydrates using spectroscopy and molecular dynamics (MD). The interaction with carbohydrates was studied by evaluating lectin fluorescence emission after titration with glucose or galactose (2.0-11 mM). The Stern-Volmer constant (Ksv), binding constant (Ka), Gibbs free energy (∆G), and Hill coefficient were calculated. After the urea-induced denaturation of cMoL, evaluations were performed using fluorescence spectroscopy, circular dichroism (CD), and hemagglutinating activity (HA) evaluations. The MD simulations were performed using the Amber 20 package. The decrease in Ksv revealed that cMoL interacts with carbohydrates via a static mechanism. The cMoL bound carbohydrates spontaneously (ΔG < 0) and presented a Ka on the order of 102, with high selectivity for glucose. Protein-ligand complexes were stabilized by hydrogen bonds and hydrophobic interactions. The Hill parameter (h~2) indicated that the binding occurs through the cMoL dimer. The loss of HA at urea concentrations at which the fluorescence and CD spectra indicated protein monomerization confirmed these results. The MD simulations revealed that glucose bound to the large cavity formed between the monomers. In conclusion, the biotechnological application of cMoL lectin requires specific methods or media to improve its dimeric protein structure.
Assuntos
Simulação de Dinâmica Molecular , Moringa oleifera , Ligação Proteica , Sementes , Moringa oleifera/química , Sementes/química , Lectinas de Plantas/química , Multimerização Proteica , Carboidratos/química , Dicroísmo Circular , Lectinas/química , Lectinas/metabolismo , Espectrometria de Fluorescência , Conformação Proteica , Termodinâmica , Ligação de HidrogênioRESUMO
AIMS: We investigated the putative fungistatic and fungicidal activities of pomegranate sarcotesta lectin (PgTeL) against Cryptococcus neoformans B3501 (serotype D), specifically the ability of PgTeL to inhibit yeast capsule and biofilm formation in this strain. METHODS AND RESULTS: PgTeL showed a minimum inhibitory concentration of 172.0 µg ml-1, at which it did not exhibit a fungicidal effect. PgTeL concentrations of 4.0-256.0 µg ml-1 reduced biofilm biomass by 31.0%-64.0%. Furthermore, 32.0-256.0 µg ml-1 PgTeL decreased the metabolic activity of the biofilm by 32.0%-93.0%. Scanning electron microscopy images clearly revealed disruption of the biofilm matrix. Moreover, PgTeL disrupted preformed biofilms. At concentrations of 8.0-256.0 µg ml-1, PgTeL reduced metabolic activity in C. neoformans by 36.0%-92.0%. However, PgTeL did not inhibit the ability of B3501 cells to form capsules under stress conditions. CONCLUSIONS: PgTeL inhibited biofilm formation and disrupted preformed biofilms, demonstrating its potential for use as an anticryptococcal agent.
Assuntos
Criptococose , Cryptococcus neoformans , Punica granatum , Lectinas/farmacologia , Punica granatum/metabolismo , Plâncton/metabolismo , Biofilmes , Testes de Sensibilidade Microbiana , Antifúngicos/farmacologia , Antifúngicos/metabolismoRESUMO
Actinomycetes are a distinct group of filamentous bacteria. The Streptomyces genus within this group has been extensively studied over the years, with substantial contributions to society and science. This genus is known for its antimicrobial production, as well as antitumor, biopesticide, and immunomodulatory properties. Therefore, the extraordinary plasticity of the Streptomyces genus has inspired new research techniques. The newest way of exploring Streptomyces has comprised the discovery of new natural metabolites and the application of emerging tools such as CRISPR technology in drug discovery. In this narrative review, we explore relevant published literature concerning the ongoing novelties of the Streptomyces genus.
Assuntos
Actinobacteria , Anti-Infecciosos , Streptomyces , Streptomyces/genética , Streptomyces/metabolismo , Biotecnologia , Anti-Infecciosos/metabolismo , Actinobacteria/metabolismoRESUMO
Despite the fact that skin has a stronger potential to regenerate than other tissues, wounds have become a serious healthcare issue. Much effort has been focused on developing efficient therapeutical approaches, especially biological ones. This paper presents a comprehensive review on the wound healing process, the classification of wounds, and the particular characteristics of each phase of the repair process. We also highlight characteristics of the normal process and those involved in impaired wound healing, specifically in the case of infected wounds. The treatments discussed here include proteins, lipids, and carbohydrates. Proteins are important actors mediating interactions between cells and between them and the extracellular matrix, which are essential interactions for the healing process. Different strategies involving biopolymers, blends, nanotools, and immobilizing systems have been studied against infected wounds. Lipids of animal, mineral, and mainly vegetable origin have been used in the development of topical biocompatible formulations, since their healing, antimicrobial, and anti-inflammatory properties are interesting for wound healing. Vegetable oils, polymeric films, lipid nanoparticles, and lipid-based drug delivery systems have been reported as promising approaches in managing skin wounds. Carbohydrate-based formulations as blends, hydrogels, and nanocomposites, have also been reported as promising healing, antimicrobial, and modulatory agents for wound management.
Assuntos
Anti-Infecciosos , Cicatrização , Animais , Pele , Lipídeos , CarboidratosRESUMO
Biomphalaria glabrata snails constitute the main vector of schistosomiasis in Brazil, and Bauhinia monandra Kurz, the leaves of which contain BmoLL lectin with biocidal action, is a plant widely found on continents in which the disease is endemic. This work describes the composition of B. monandra preparations and the effect on embryos and adult snails, their reproduction parameters and hemocytes. We also describe the results of a comet assay after B. glabrata exposure to sublethal concentrations of the preparations. Additionally, the effects of the preparations on S. mansoni cercariae and environmental monitoring with Artemia salina are described. In the chemical evaluation, cinnamic, flavonoid and saponin derivatives were detected in the two preparations assessed, namely the saline extract and the fraction. Both preparations were toxic to embryos in the blastula, gastrula, trochophore, veliger and hippo stages (LC50 of 0.042 and 0.0478; 0.0417 and 0.0419; 0.0897 and 0.1582; 0.3734 and 0.0974; 0.397 and 0.0970 mg/mL, respectively) and to adult snails (LC50 of 6.6 and 0.87 mg/mL, respectively), which were reproductively affected with decreased egg deposition. In blood cell analysis, characteristic cells for apoptosis, micronucleus and binucleation were detected, while for comet analysis, different degrees of nuclear damage were detected. The fraction was able to cause total mortality of the cercariae and did not present environmental toxicity. Therefore, B. monandra preparations are promising in combating schistosomiasis since they can control both the intermediate host and eliminate the infectious agent, besides being safe to the environment.
Assuntos
Bauhinia , Biomphalaria , Esquistossomose , Animais , Artemia , Folhas de Planta , Schistosoma mansoniRESUMO
Usnic acid is the best-studied lichen metabolite, presenting several biological activities, such as antibacterial, immunostimulating, antiviral, antifungal, anti-inflammatory, and antiparasitic agents; despite these relevant properties, it is a hydrophobic and toxic molecule. In this context, scientific research has driven the development of innovative alternatives, considering usnic acid as a source of raw material in obtaining new molecules, allowing structural modifications (syntheses) from it. The purpose is to optimize biological activities and toxicity, with less concentration and/or response time. This work presents a literature review with an analogy of the hydrophobic molecule of usnic acid with its hydrophilic derivative of potassium usnate, emphasizing the elucidation and structural characteristics, biological activities, and toxicological aspects of both molecules, and the advantages of using the promising derivative hydrophilic in different in vitro and in vivo assays when compared to usnic acid.
Assuntos
Benzofuranos/química , Benzofuranos/farmacologia , Potássio/química , Analgésicos/química , Analgésicos/farmacologia , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Antiparasitários/química , Antiparasitários/farmacologia , Benzofuranos/toxicidade , Interações Hidrofóbicas e Hidrofílicas , Líquens/metabolismoRESUMO
The partitioning and purification of lectins from the crude extract of Cratylia mollis seeds (Cramoll 1,4) was investigated in aqueous two-phase systems (ATPS). A factorial design model (24) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1500-8000 g/mol), PEG concentration (12.5-17.5% w/w), phosphate (10-15% w/w) concentration, and pH (6-8) on the differential partitioning, purification factor, and yield of the lectin. Polymer and salt concentration were the most important variables affecting partition of lectin and used to find optimum purification factor by experimental Box-Behnken design together with the response surface methodology (RSM). ATPS showed best conditions composed by 13.9% PEG1500, 15.3% phosphate buffer at pH 6, which ensured purification factor of 4.70. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band of protein with 26.1 kDa. Furthermore, results demonstrated a thermostable lectin presenting activity until 60 °C and lost hemagglutinating activity at 80 °C. According to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of lectins.
Assuntos
Lectinas/química , Lectinas/isolamento & purificação , Phaseolus/química , Extratos Vegetais/química , Eletroforese em Gel de Poliacrilamida , Hemaglutininas/química , Concentração de Íons de Hidrogênio , Fosfatos/química , Polietilenoglicóis/química , Proteínas/química , Sementes/química , Espectrofotometria , Propriedades de Superfície , TemperaturaRESUMO
The genus Aeromonas comprises more than thirty Gram-negative bacterial species which mostly act as opportunistic microorganisms. These bacteria are distributed naturally in diverse aquatic ecosystems, where they are easily isolated from animals such as fish and crustaceans. A capacity for adaptation also makes Aeromonas able to colonize terrestrial environments and their inhabitants, so these microorganisms can be identified from different sources, such as soils, plants, fruits, vegetables, birds, reptiles, amphibians, among others. Infectious processes usually develop in immunocompromised humans; in fish and other marine animals this process occurs under conditions of stress. Such events are most often associated with incorrect practices in aquaculture. Aeromonas has element diverse ranges, denominated virulence factors, which promote adhesion, colonization and invasion into host cells. These virulence factors, such as membrane components, enzymes and toxins, for example, are differentially expressed among species, making some strains more virulent than others. Due to their diversity, no single virulence factor was considered determinant in the infectious process generated by these microorganisms. Unlike other genera, Aeromonas species are erroneously differentiated by conventional biochemical tests. Therefore, molecular assays are necessary for this purpose. Nevertheless, new means of identification have been considered in order to generate methods that, like molecular tests, can correctly identify these microorganisms. The main objectives of this review are to explain environmental and structural characteristics of the Aeromonas genus and to discuss virulence mechanisms that these bacteria use to infect aquatic organisms and humans, which are important aspects for aquaculture and public health, respectively. In addition, this review aims to clarify new tests for the precise identification of the species of Aeromonas, contributing to the exact and specific diagnosis of infections by these microorganisms and consequently the treatment.
Assuntos
Aeromonas/classificação , Aeromonas/isolamento & purificação , Doenças dos Animais/microbiologia , Microbiologia Ambiental , Infecções por Bactérias Gram-Negativas/veterinária , Doenças das Plantas/microbiologia , Aeromonas/patogenicidade , Animais , Técnicas Bacteriológicas/métodos , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Técnicas de Diagnóstico Molecular/métodos , Virulência , Fatores de Virulência/genéticaRESUMO
Aedes aegypti control is a key component of the prophylaxis of dengue fever and other diseases. Moringa oleifera seeds contain a water-soluble lectin (WSMoL) with larvicidal and ovicidal activities against this insect. In this study, A. aegypti individuals were exposed at the third larval instar for 24â¯h to the water extract (0.1-1.0â¯mg/mL of protein) or lectin-rich fraction (0.05-0.6â¯mg/mL of protein) containing WSMoL, and then their survival and development were followed for 9 days post-exposure. The feeding capacity of adult females that developed from the treated larvae and the hatching success of eggs laid by them were also evaluated. Further, any alterations to the midgut histology of treated larvae, pupae, and adults were investigated. The extract and fraction induced the death of A. aegypti larvae along the post-exposure period. Both preparations also delayed the developmental cycle. The midguts of treated larvae and pupae showed disorganization and epithelial vacuolization, while in treated adults, the epithelium was underdeveloped compared to control. Unlike in control mosquitos, proliferating cells were not detected in treated larvae, and appeared in lower numbers in treated pupae than in control pupae. Adult females that developed from larvae treated with the fraction gained less weight after a blood meal compared with control. The amount of eggs laid by females that developed from larvae treated with both the extract and fraction was significantly lower than in control. In addition, the eggs showed lower hatching rates. In conclusion, females that developed from larvae treated with both the water extract and lectin-rich fraction showed reduced engorgement after a blood meal, with the consequent impairment of their fertility and fecundity. These results were probably due to the damage to midgut organization and impairment of the remodeling process during metamorphosis.
Assuntos
Aedes/efeitos dos fármacos , Lectinas/farmacologia , Moringa oleifera/química , Extratos Vegetais/farmacologia , Aedes/crescimento & desenvolvimento , Aedes/fisiologia , Animais , Feminino , Fertilidade/efeitos dos fármacos , Inseticidas/química , Inseticidas/farmacologia , Intestinos/efeitos dos fármacos , Intestinos/crescimento & desenvolvimento , Intestinos/patologia , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lectinas/química , Controle de Mosquitos , Extratos Vegetais/química , Pupa/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Pupa/fisiologia , Sementes/química , Água/químicaRESUMO
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⯵g/mL, respectively. In addition, it showed bactericidal effect on the non-resistant isolate (minimal bactericidal concentration: 200⯵g/mL). For Candida albicans and Candida parapsilosis, ApuL showed fungistatic effect (MIC50: 200 and 400⯵g/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.
Assuntos
Alpinia/química , Antibacterianos/farmacologia , Antifúngicos/farmacologia , Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Lectinas/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Candida albicans/crescimento & desenvolvimento , Candida albicans/fisiologia , Sinergismo Farmacológico , Lectinas/isolamento & purificação , Testes de Sensibilidade Microbiana , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/fisiologiaRESUMO
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.
Assuntos
Compostos Azo/química , Corantes/química , Metaloporfirinas/química , Redes Neurais de Computação , Ácidos Sulfanílicos/química , Poluentes Químicos da Água/química , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/química , Ferro/química , Lactuca/efeitos dos fármacos , Oxirredução , Fotólise , Salmonella enteritidis/efeitos dos fármacos , Luz Solar , Poluentes Químicos da Água/toxicidadeRESUMO
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%.
Assuntos
Corantes/química , Peróxido de Hidrogênio/química , Redes Neurais de Computação , Corantes/toxicidade , Humanos , Cinética , Oxirredução , Estresse Oxidativo , Folhas de Planta , Staphylococcus/efeitos dos fármacos , Raios Ultravioleta , Purificação da ÁguaRESUMO
Biotechnological techniques allow the investigation of alternatives to outdated chemical insecticides for crop protection; some investigations have focused on the identification of molecules tailored from nature for this purpose. We, herein, describe the negative effects of water-soluble lectin from Moringa oleifera seeds (WSMoL) on Anagasta kuehniella development. The chitin-binding lectin, WSMoL, impaired the larval weight gain by 50% and affected the activity of the pest's major digestive enzymes. The commitment of the digestive process became evident after controlled digestion studies, where the capacity of protein digestion was compromised by >90%. Upon acute exposure, the lectin was not resistant to digestion; however, chronic ingestion of WSMoL was able to reverse this feature. Thus, we show that resistance to digestion may not be a prerequisite for a lectin's ability to exert negative effects on larval physiology. The mechanism of action of WSMoL involves binding to chitin with possible disruption to the peritrophic membrane, causing disorder between the endo- and ectoperitrophic spaces. Additionally, results suggest that WSMoL may trigger apoptosis in gut cells, leading to the lower enzymatic activity observed in WSMoL-fed larvae. Although assays employing an artificial diet did not demonstrate effects of WSMoL on A. kuehniella mortality, this lectin may hold potential for exerting insecticide effects on other pest insects, as well for use in other experimental approaches, such as WSMoL-expressing plants. Moreover, the use of WSMoL with other biotechnological tools, such as 'pyramid' crops, may represent a strategy for delaying the evolution of pest resistance to transgenic crops, since its multiple site targets could act in synergism with other insecticide compounds.
Assuntos
Inseticidas/farmacologia , Lectinas/farmacologia , Moringa oleifera/química , Mariposas/fisiologia , Animais , Quitina/química , Fenômenos Fisiológicos do Sistema Digestório/efeitos dos fármacos , Inseticidas/química , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/fisiologia , Lectinas/química , Mariposas/efeitos dos fármacos , Mariposas/crescimento & desenvolvimento , Sementes/químicaRESUMO
Moringa oleifera flower extract, with trypsin inhibitor activity, is a larvicidal agent on Aedes aegypti. This work reports the isolation of trypsin inhibitor (M. oleifera flower trypsin inhibitor (MoFTI)) and its effect on A. aegypti egg hatching, viability of newly hatched larvae, survival of pupae, and growth of inhabitant bacteria from midgut of fourth-instar larvae (L4). MoFTI (K i, 2.4 µM), isolated by affinity chromatography on trypsin-agarose column, was an 18.2 kDa polypeptide on sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Flower extract (at concentrations of 8.5-17.0 mg/mL) reduced egg hatchability while MoFTI (0.05-0.5 mg/mL) did not affect the hatching rate. Mortality of newly hatched larvae ranged from 3.5 to 19.1 % in the presence of the extract (4.0-17.0 mg/mL) and was also promoted by MoFTI (LC50, 0.3 mg/mL). After 72 h, larvae incubated with extract at 13.0 and 17.0 mg/mL were at stages L2 and L1, respectively, while in control they reached L3 instar. In the presence of MoFTI, at all concentrations tested, the larvae did not pass the first instar. Flower extract and MoFTI did not interfere on pupae survival. The extract and MoFTI inhibited the growth of L4 gut bacteria (minimum inhibitory concentrations of 3.47 and 0.031 mg/mL, respectively) but only the inhibitor showed bactericide effect (minimum bactericidal concentration of 1.0 mg/mL). The findings reported herein indicate that MoFTI constitutes a larvicidal principle from M. oleifera flowers against A. aegypti newly hatched larvae and is an antibacterial agent active against the microbiota from L4 gut.
Assuntos
Aedes/crescimento & desenvolvimento , Aedes/microbiologia , Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Inseticidas/farmacologia , Moringa oleifera/química , Inibidores da Tripsina/farmacologia , Aedes/efeitos dos fármacos , Animais , Antibacterianos/isolamento & purificação , Sistema Digestório/microbiologia , Flores/química , Inseticidas/análise , Inseticidas/isolamento & purificação , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/microbiologia , Óvulo/efeitos dos fármacos , Óvulo/fisiologia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Pupa/efeitos dos fármacos , Pupa/crescimento & desenvolvimento , Tripsina/metabolismo , Inibidores da Tripsina/isolamento & purificaçãoRESUMO
The water-soluble lectin isolated from Moringa oleifera seeds (WSMoL) is a larvicidal, ovicidal, and oviposition-stimulating agent against Aedes aegypti under laboratory conditions. This study investigated the effect of WSMoL in traps for the capture of A. aegypti eggs and adult females under semi-field conditions and determined whether gravid females could detect WSMoL by an olfactory response. WSMoL was isolated according to a previously described procedure using chitin chromatography. The bioassays were performed in large cages (12.5 m(3)). Two traps for collection of eggs (ovitrap) or adult mosquitoes (MosquiTRAP(TM)) were placed in a cage. One was filled with WSMoL (0.1 mg/mL) and the other with tap water (negative control). An infusion of Panicum maximum leaves was used as a positive control. Forty gravid females were then released in each cage. After 2 (for oviposition) or 3 h (for female capture), the traps were removed, and the number of eggs or females was counted. An olfactometry assay was performed to investigate whether the effect of WSMoL on gravid females was linked to an olfactory response. WSMoL showed an oviposition-stimulating effect (65 ± 14%) that was similar (p < 0.05) to that promoted by the P. maximum infusion (67 ± 11%). The efficiency of MosquiTRAP(TM) in capturing gravid females was not increased by WSMoL. The olfactometry assay indicated that the response of females to WSMoL did not involve the stimulation of olfactory sensilla. WSMoL effectively captured eggs when used in ovitraps under semi-field conditions; this property, together with the ovicidal and larvicidal activities of this lectin, makes it an interesting candidate for A. aegypti control.
Assuntos
Aedes/fisiologia , Lectinas/farmacologia , Moringa oleifera/química , Oviposição/efeitos dos fármacos , Sementes/química , Animais , Feminino , Controle de Mosquitos , Óvulo/efeitos dos fármacos , OlfatoRESUMO
The indiscriminate use of synthetic insecticides to control Aedes aegypti has led to emergence of resistant populations. Moringa oleifera seeds contain the lectins WSMoL and cMoL. WSMoL has larvicidal activity on fourth-stage of A. aegypti organophosphate-susceptible larvae (Rockefeller L4). This study reports on the effects of cMoL on the survival of Rockefeller L4 as well as of WSMoL and cMoL on L4 from an organophosphate-resistant population (Rec-R). The effects of lectins on digestive (amylase, trypsin, and protease) and detoxifying (superoxide dismutase (SOD), α- and ß-esterases) enzymes from larvae were also determined. cMoL (0.1-0.8 mg/ml) did not kill Rockefeller L4 as well as WSMoL and cMoL (0.1-0.8 mg/ml) were not larvicidal for Rec-R L4. WSMoL stimulated protease, trypsin-like, and α-amylase from Rockefeller L4 while cMoL inhibited these enzymes. WSMoL had no effect on trypsin-like activity from Rec-R L4 but inhibited protease and α-amylase. Among digestive enzymes of Rec-R L4, cMoL inhibited only trypsin-like activity. cMoL inhibited SOD activities from Rockefeller and Rec-R L4 in a higher level than WSMoL while ß-esterase from Rockefeller L4 was more inhibited by WSMoL. The lectins promoted low stimulation or inhibition of α-esterase activities from both populations. In conclusion, Rockefeller and Rec-R larvae were distinctly affected by M. oleifera lectins, and larvicidal mechanism of WSMoL on Rockefeller L4 may involve deregulation of digestive enzymes. cMoL interfered mainly on SOD activity and thus it can be investigated as a synergistic agent for controlling populations whose resistance is linked to an increased detoxifying process mediated by this enzyme.
Assuntos
Aedes/efeitos dos fármacos , Inseticidas , Moringa oleifera/química , Lectinas de Plantas/farmacologia , Aedes/enzimologia , Animais , Esterases/metabolismo , Resistência a Inseticidas , Larva/efeitos dos fármacos , Larva/enzimologia , Organofosfatos , Extratos Vegetais/farmacologia , Sementes/química , Superóxido Dismutase/metabolismoRESUMO
Environmental factors such as seasonal cycles are the main chronic stress cause in fish increasing incidence of disease and mortality and affecting productive performance. Arapaima gigas (pirarucu) is an Amazonian air-breathing and largest freshwater fish with scales in the world. The captivity development of pirarucu is expanding since it can fatten up over 1 kg per month reaching 10 kg body mass in the first year of fattening. This work was conducted in three periods (April to July 2010, August to November 2010, and December 2010 to March 2011) defined according to rainfall and medium temperatures. Seasonality effect analysis was performed on biochemical (lectin activity, lactate dehydrogenase, and alkaline phosphatase activities) and hematological (total count of red blood cells, hematocrit, hemoglobin, and hematimetric Wintrobe indexes) stress indicators, as well as on growth and wellbeing degree expressed by pirarucu condition factor developed in captivity. All biochemical and hematological stress indicators showed seasonal variations. However, the fish growth was allometrically positive; condition factor high values indicated good state of healthiness in cultivation. These results reinforce the robust feature of pirarucu and represent a starting point for understanding stress physiology and environmental changes during cultivation enabling identification and prevention of fish adverse health conditions.
Assuntos
Fosfatase Alcalina/sangue , Proteínas de Peixes/sangue , Peixes/sangue , L-Lactato Desidrogenase/sangue , Estações do Ano , Estresse Fisiológico , AnimaisRESUMO
WSMoL, a water-soluble lectin from the seeds of Moringa oleifera, present several biological activities. This work aimed to evaluated the toxicity and antitumor activity of WSMoL. To analyze toxicity, it was determined hematological, biochemical and histological parameters; consumption of water and feed as well as the weight of the animals. Antitumor analysis included evaluation of tumor weight, histology and cytokine levels. Acute toxicity assay revealed 60% mortality of animals treated with lectin at 200 mg/kg i. p. At 100 mg/kg i. p., the animals showed a decreased food and water consumption as well weight gain in comparison with control. However, no animal died and there were no alterations in blood parameters or histological analysis. Antitumor activity evaluated at safe doses (2.5, 5 and 10 mg/kg) showed a significant reduction in tumor weight. Tumor photomicrographs evidenced that WSMoL treatment reduced dissemination of tumor cells. WSMoL (5 and 10 mg/kg) significantly enhance the immune function in the tumor environment as showed by increased the levels of pro-inflammatory (TNF-α, IFN-γ, IL-2, IL-6, and IL-17) and anti-inflammatory (IL-4 and IL-10) cytokines. In conclusion, WSMoL showed in vivo antitumor activity in mice bearing sarcoma 180 tumor, probably by increase the immune response against the tumor.
Assuntos
Moringa oleifera , Sarcoma 180 , Animais , Camundongos , Lectinas , Água , Sarcoma 180/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Extratos Vegetais/toxicidade , Citocinas , SementesRESUMO
Triturated Moringa oleifera seeds have components that adsorb recalcitrant indigo carmine dye. Coagulating proteins known as lectins (carbohydrate-binding proteins) have already been purified from the powder of these seeds, in milligram amounts. The coagulant lectin from M. oleifera seeds (cMoL) was characterized by potentiometry and scanning electron microscopy (SEM) using MOFs, or metal-organic frameworks, of [Cu3(BTC)2(H2O)3]n to immobilize cMoL and construct biosensors. The potentiometric biosensor revealed an increase in the electrochemical potential resulting from the Pt/MOF/cMoL interaction with different concentrations of galactose in the electrolytic medium. The developed aluminum batteries constructed with recycled cans degraded an indigo carmine dye solution; the oxide reduction reactions of the batteries generated Al(OH)3, promoting dye electrocoagulation. Biosensors were used to investigate cMoL interactions with a specific galactose concentration and monitored residual dye. SEM revealed the components of the electrode assembly steps. Cyclic voltammetry showed differentiated redox peaks related to dye residue quantification by cMoL. Electrochemical systems were used to evaluate cMoL interactions with galactose ligands and efficiently degraded dye. Biosensors could be used for lectin characterization and monitoring dye residues in environmental effluents of the textile industry.
Assuntos
Lectinas , Moringa oleifera , Lectinas/análise , Moringa oleifera/química , Índigo Carmim/análise , Galactose , Sementes/química , Carmim/análiseRESUMO
The association of quantum dots (QDs) to carbohydrate-binding proteins - lectins - has revealed novel biotechnological strategies for glycobiology studies. Herein, carboxyl-coated QDs were conjugated by adsorption to Cramoll, a glucose/mannose lectin obtained from Cratylia mollis seeds. Then, the conjugates were optically characterized and used to evaluate the surface carbohydrate profiles of four Aeromonas species isolated from the tambaqui fish (Colossoma macropomum). All the Aeromonas cells were labeled by the conjugate. Inhibition assays with methyl-α-D-mannopyranoside and mannan were performed to confirm the labeling specificity. Cramoll-QDs conjugates presented high brightness and showed similar absorption and emission profiles compared to bare QDs. According to the labeling pattern of Aeromonas spp. by the conjugate, results suggested that A. jandaei and A. dhakensis strains may harbor a higher content of more complex glucose/mannose surface glycans, with more available sites for Cramoll-QDs interaction, than A. hydrophila and A. caviae. Noteworthy, the Cramoll-QDs conjugates demonstrated to be potential tools for bacterial characterization based on superficial carbohydrate detection.