RESUMO
Terpenoids, also named terpenes or isoprenoids, are a family of natural products found in all living organisms. Many plants produce terpenoids as secondary metabolites, and these make up a large part of essential oils. One of most important characteristic is that the compounds are volatile, have odor and can be used in a variety of applications in different industrial segments and traditional medicine. Brazil has a rich and diverse flora that can be used as a source of research for obtaining new molecules. Within the Brazilian flora, it is worth mentioning the Caatinga as an exclusively Brazilian biome where plants adapt to a specific series of weather conditions and therefore become a great storehouse of the terpenoid compounds to be described herein. Fungal infections have become increasingly common, and a great demand for new agents with low toxicity and side effects has thus emerged. Scientists must search for new molecules exhibiting antifungal activity to develop new drugs. This review aims to analyze scientific data from the principal published studies describing the use of terpenes and their biological applications as antifungals.
Assuntos
Óleos Voláteis , Terpenos , Terpenos/farmacologia , Terpenos/metabolismo , Antifúngicos/farmacologia , Brasil , Óleos Voláteis/farmacologia , PlantasRESUMO
Cryptococcosis is an opportunistic systemic mycosis whose treatment is limited to three drugs. In this work, we evaluated the antifungal activity of a hexane extract (HE) from Spondias tuberosa leaves against Cryptococcus neoformans and Cryptococcus gattii. Minimal inhibitory concentrations (MIC) were determined, and putative mechanisms were evaluated by flow cytometry. In addition, an in vivo infection assay was performed using Tenebrio molitor larvae. Treatment with HE inhibited the growth of standard and clinical isolates of C. neoformans and C. gattii (MICs ranging from 0.78 to 3.12mg/mL), significantly (P<0.05) increased mitochondrial superoxide anion levels, and induced mitochondrial membrane depolarization, loss of lysosomal membrane integrity, and phosphatidylserine externalization. The mean survival time of C. gattii-infected T. molitor larvae significantly (P<0.05) increased from 1.225 days in control to 3.067 and 3.882 days in HE-treated groups (78 and 156mg/kg, respectively). In conclusion, HE showed anticryptococcal activity, induced mitochondrial and lysosomal damage in yeast cells, and exhibited anti-infective action against C. gattii in T. molitor larvae.
Assuntos
Anacardiaceae/química , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Criptococose/tratamento farmacológico , Hexanos/química , Animais , Antifúngicos/efeitos adversos , Antifúngicos/uso terapêutico , Criptococose/patologia , Cryptococcus gattii/citologia , Cryptococcus gattii/efeitos dos fármacos , Cryptococcus gattii/ultraestrutura , Cryptococcus neoformans/citologia , Cryptococcus neoformans/efeitos dos fármacos , Cryptococcus neoformans/ultraestrutura , Hexanos/farmacologia , Humanos , Larva/efeitos dos fármacos , Lisossomos/efeitos dos fármacos , Lisossomos/fisiologia , Testes de Sensibilidade Microbiana , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , Fitoterapia , Extratos Vegetais/química , Tenebrio/efeitos dos fármacos , Tenebrio/crescimento & desenvolvimento , Testes de ToxicidadeRESUMO
The resistance of micro-organisms to antimicrobial agents has been a challenge to treat animal and human infections, and for environmental control. Lectins are natural proteins and some are potent antimicrobials through binding to carbohydrates on microbial surfaces. Oligomerization state of lectins can influence their biological activity and maximum binding capacity; the association among lectin polypeptide chains can alter the carbohydrate-lectin binding dissociation rate constants. Antimicrobial mechanisms of lectins include the pore formation ability, followed by changes in the cell permeability and latter, indicates interactions with the bacterial cell wall components. In addition, the antifungal activity of lectins is associated with the chitin-binding property, resulting in the disintegration of the cell wall or the arrest of de novo synthesis from the cell wall during fungal development or division. Quorum sensing is a cell-to-cell communication process that allows interspecies and interkingdom signalling which coordinate virulence genes; antiquorum-sensing therapies are described for animal and plant lectins. This review article, among other approaches, evaluates lectins as antimicrobials.
Assuntos
Anti-Infecciosos/farmacologia , Bactérias/efeitos dos fármacos , Resistência Microbiana a Medicamentos , Fungos/efeitos dos fármacos , Lectinas/farmacologia , Animais , Infecções Bacterianas/microbiologia , Humanos , Micoses/microbiologia , Percepção de Quorum/efeitos dos fármacosRESUMO
Zymomonas mobilis has long attracted attention owing to its capacity to ferment hexose to ethanol. From a taxonomic viewpoint, Z. mobilis is a unique species of the genus Zymomonas, separated into three subspecies, Z. mobilis subsp. mobilis, Z. mobilis subsp. pomaceae and Z. mobilis subsp. francensis on the basis of physiological tests, which are often unreliable owing to the genetic proximity among these species. Currently, the use of molecular techniques is more appropriate for identification of these bacterial subspecies. In this study, the 32 strains of Z. mobilis present in the UFPEDA bacterial collection were characterized using molecular techniques, such as sequencing of the 16S rDNA gene and its theoretical restriction profile, classifying them as members of the subspecies, Z. mobilis subsp. mobilis. In addition, anaerobic cultivations were performed, which showed the biological diversity of the strains in terms of growth, sugar consumption and ethanol production. From these results, it was possible to identify the strain Z-2-80 as a promising bacterium for use in the fermentation process. SIGNIFICANCE AND IMPACT OF THE STUDY: Zymomonas mobilis is a bacterium of great relevance to biotechnology, owing to its capacity to ferment hexose to ethanol. On a molecular basis, 32 isolates were identified as Z. mobilis subsp. mobilis. However, intraspecific diversity was identified when these were grown under strictly anaerobic conditions. The results obtained from this study suggest a strain of Z. mobilis as an alternative for use in the fermentation process.
Assuntos
Reatores Biológicos/microbiologia , DNA Bacteriano/genética , Etanol/metabolismo , Zymomonas/classificação , Zymomonas/metabolismo , Anaerobiose , Brasil , DNA Ribossômico/genética , Fermentação , Hexoses/metabolismo , RNA Ribossômico 16S/genética , Zymomonas/genética , Zymomonas/isolamento & purificaçãoRESUMO
BACKGROUND: Quantum dots (QDs) are outstanding nanomaterials of great interest to life sciences. Their conjugation versatility added to unique optical properties, highlight these nanocrystals as very promising fluorescent probes. Among uncountable new nanosystems, in the last years, QDs conjugated to glycans or lectins have aroused a growing attention and their application as a tool to study biological and functional properties has increased. SCOPE OF REVIEW: This review describes the strategies, reported in the literature, to conjugate QDs to lectins or carbohydrates, providing valuable information for the elaboration, improvement, and application of these nanoconjugates. It also presents the main applications of these nanosystems in glycobiology, such as their potential to study microorganisms, the development of diseases such as cancer, as well as to develop biosensors. MAJOR CONCLUSIONS: The development of glyconanoparticles based on QDs emerged in the last decade. Many works reporting the conjugation of QDs with carbohydrates and lectins have been published, using different strategies and reagents. These bioconjugates enabled studies that are very sensitive and specific, with potential to detect and elucidate the glycocode expressed in various normal or pathologic conditions. GENERAL SIGNIFICANCE: Produce a quick reference source over the main advances reached in the glyconanotechnology using QDs as fluorescent probes.
Assuntos
Glicoconjugados , Nanotecnologia/métodos , Pontos Quânticos , Técnicas Bacteriológicas , Técnicas Biossensoriais , Metabolismo dos Carboidratos , Carboidratos/análise , Linhagem Celular Tumoral , Técnicas de Química Analítica/métodos , Fluorescência , Glicoconjugados/administração & dosagem , Glicoconjugados/química , Glicoconjugados/uso terapêutico , Humanos , Lectinas/administração & dosagem , Lectinas/química , Nanopartículas Metálicas , Modelos Moleculares , Micologia/métodos , Nanotecnologia/tendências , Neoplasias/química , Neoplasias/diagnóstico , Imagem Óptica/métodos , Parasitologia/métodos , Pontos Quânticos/administração & dosagem , Pontos Quânticos/química , Pontos Quânticos/uso terapêuticoRESUMO
Infections are one of the main reasons for removal of implants from patients, and usually need difficult and expensive treatments. Staphylococcus aureus and Staphylococcus epidermidis are the most frequently detected pathogens. We reviewed the epidemiology and pathogenesis of implant-related infections. Relevant studies were identified by electronic searching of the following databases: PubMed, ScienceDirect, Academic Google, and CAPES Journal Portal. This review reports epidemiological studies of implant infections caused by S. aureus and S. epidermidis. We discuss some methodologies used in the search for new compounds with antibiofilm activity and the main strategies for biomaterial surface modifications to avoid bacterial plaque formation and consequent infection. S. aureus and S. epidermidis are frequently involved in infections in catheters and orthopaedic/breast implants. Different methodologies have been used to test the potential antibiofilm properties of compounds; for example, crystal violet dye is widely used for in-vitro biofilm quantification due to its low cost and good reproducibility. Changes in the surface biomaterials are necessary to prevent biofilm formation. Some studies have investigated the immobilization of antibiotics on the surfaces of materials used in implants. Other approaches have been used as a way to avoid the spread of bacterial resistance to antimicrobials, such as the functionalization of these surfaces with silver and natural compounds, as well as the electrical treatment of these substrates.
Assuntos
Infecções Relacionadas à Prótese/epidemiologia , Infecções Relacionadas à Prótese/microbiologia , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificação , Staphylococcus epidermidis/isolamento & purificação , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Infecções Relacionadas a Cateter/epidemiologia , Infecções Relacionadas a Cateter/microbiologia , Materiais Revestidos Biocompatíveis , Humanos , Staphylococcus aureus/fisiologia , Staphylococcus epidermidis/fisiologia , Propriedades de SuperfícieRESUMO
Fusarium wilt caused by Fusarium oxysporum f. sp lycopersici (Fol) is one of the main diseases affecting tomatoes. The BHRS 2,3 genotype of tomato is, however, resistant to this disease. A proteomic approach was used to understand the defense mechanisms of this genotype using the tomato root, the first tissue that interacts with the fungus, as a target. Protein was extracted and separated by two-dimensional electrophoresis followed by staining with Coomassie brilliant blue. The proteins were identified by MALDI-TOF/TOF mass spectrometry. A total of 22 proteins were identified, 21 of which showed differential expression with 12 proteins being upregulated and nine being downregulated. Plants responded to the pathogen with increased expression of pathogenesis-related proteins. We noted the induction of proteins involved in hypersensitivity reaction and other defense mechanisms. The expression of proteins of primary metabolism related to energy production, however, decreased, as did the expression of two proteins related to defense against abiotic stress. These results demonstrate the presence of important mechanisms for defense against Fol in the tomato genotype BHRS 2,3.
Assuntos
Resistência à Doença/genética , Fusarium/patogenicidade , Raízes de Plantas/metabolismo , Proteoma/genética , Solanum lycopersicum/genética , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/imunologia , Solanum lycopersicum/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/microbiologia , Proteoma/metabolismo , Estresse FisiológicoRESUMO
In the present study, we compared the performance of a ThinPrep cytological method with the conventional Papanicolaou test for diagnosis of cytopathological changes, with regard to unsatisfactory results achieved at the Central Public Health Laboratory of the State of Pernambuco. A population-based, cross-sectional study was performed with women aged 18 to 65 years, who spontaneously sought gynecological services in Public Health Units in the State of Pernambuco, Northeast Brazil, between April and November 2011. All patients in the study were given a standardized questionnaire on sociodemographics, sexual characteristics, reproductive practices, and habits. A total of 525 patients were assessed by the two methods (11.05% were under the age of 25 years, 30.86% were single, 4.4% had had more than 5 sexual partners, 44% were not using contraception, 38.85% were users of alcohol, 24.38% were smokers, 3.24% had consumed drugs previously, 42.01% had gynecological complaints, and 12.19% had an early history of sexually transmitted diseases). The two methods showed poor correlation (k=0.19; 95%CI=0.11-0.26; P<0.001). The ThinPrep method reduced the rate of unsatisfactory results from 4.38% to 1.71% (χ2=5.28; P=0.02), and the number of cytopathological changes diagnosed increased from 2.47% to 3.04%. This study confirmed that adopting the ThinPrep method for diagnosis of cervical cytological samples was an improvement over the conventional method. Furthermore, this method may reduce possible losses from cytological resampling and reduce obstacles to patient follow-up, improving the quality of the public health system in the State of Pernambuco, Northeast Brazil.
Assuntos
Teste de Papanicolaou/métodos , Doenças do Colo do Útero/diagnóstico , Adulto , Idoso , Brasil , Estudos Transversais , Método Duplo-Cego , Feminino , Humanos , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Fatores Socioeconômicos , Adulto JovemRESUMO
Previous results provided evidence that Cratylia mollis seed lectin (Cramoll 1,4) promotes Trypanosoma cruzi epimastigotes death by necrosis via a mechanism involving plasma membrane permeabilization to Ca(2+) and mitochondrial dysfunction due to matrix Ca(2+) overload. In order to investigate the mechanism of Ca(2+) -induced mitochondrial impairment, experiments were performed analyzing the effects of this lectin on T. cruzi mitochondrial fraction and in isolated rat liver mitochondria (RLM), as a control. Confocal microscopy of T. cruzi whole cell revealed that Cramoll 1,4 binding to the plasma membrane glycoconjugates is followed by its internalization and binding to the mitochondrion. Electrical membrane potential (∆Ψm ) of T. cruzi mitochondrial fraction suspended in a reaction medium containing 10 µM Ca(2+) was significantly decreased by 50 µg/ml Cramoll 1,4 via a mechanism insensitive to cyclosporine A (CsA, membrane permeability transition (MPT) inhibitor), but sensitive to catalase or 125 mM glucose. In RLM suspended in a medium containing 10 µM Ca(2+) this lectin, at 50 µg/ml, induced increase in the rate of hydrogen peroxide release, mitochondrial swelling, and ∆Ψm disruption. All these mitochondrial alterations were sensitive to CsA, catalase, and EGTA. These results indicate that Cramoll 1, 4 leads to inner mitochondrial membrane permeabilization through Ca(2+) dependent mechanisms in both mitochondria. The sensitivity to CsA in RLM characterizes this lectin as a MPT inducer and the lack of CsA effect identifies a CsA-insensitive MPT in T. cruzi mitochondria.
Assuntos
Ciclosporina/farmacologia , Fabaceae/química , Lectinas/farmacologia , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias/metabolismo , Sementes/química , Animais , Transporte Biológico/efeitos dos fármacos , Cálcio/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Ratos , Espécies Reativas de Oxigênio/metabolismo , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/metabolismoRESUMO
Few chronic food protein models have described the relationship between allergenicity and the molecular structure of food protein after physical processing. The effect of γ-radiation on the structure of food protein was measured by fluorescence, circular dichroism and microcalorimetry. BALB/c mice were intraperitoneally sensitized and then given non-irradiated and irradiated Con-A by daily gavage for 28days. The tendency to form insoluble amorphous aggregates and partially unfolded species was observed after irradiation. The administration of non-irradiated and irradiated samples at low-dose significantly increased weight loss as well as plasma levels of eotaxin in animals repeatedly exposed to Con-A. Significant lymphocytic infiltrate filling completely the stroma of microvilli and tubular glands was observed in the small intestinal of the group given Con-A irradiated at a low dose. This phenotype was not observed in animals treated with Con-A irradiated at a high dose.
Assuntos
Concanavalina A/química , Concanavalina A/imunologia , Concanavalina A/efeitos da radiação , Hipersensibilidade Alimentar/etiologia , Administração Oral , Animais , Varredura Diferencial de Calorimetria , Dicroísmo Circular , Concanavalina A/administração & dosagem , Modelos Animais de Doenças , Relação Dose-Resposta à Radiação , Feminino , Hipersensibilidade Alimentar/patologia , Raios gama , Intestino Delgado/imunologia , Intestino Delgado/patologia , Linfócitos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Microvilosidades/imunologia , Microvilosidades/patologia , Conformação Proteica , Redução de PesoRESUMO
Libidibia ferrea has been used in folk medicine throughout Brazil, and this study evaluated the biological activities of crude extract (CE) as well as a partially purified fraction (F80) obtained from its pods. Results from the MTT assay revealed that only F80 inhibited NCI-H292 cell growth; however, neither CE nor F80 reduced HEp-2 cell growth or sarcoma 180 tumor weight with the in vivo assay. Acute oral toxicity of the extract and fraction was evaluated following the steps of Guideline 423, using female mice; LD(50) for both preparations was determined as 2,500 mg/kg body weight. CE and F80 promoted a reduction of the leukocyte number and nitrite level in inflammatory exudates when the anti-inflammatory assay (carrageenan-induced peritonitis) was performed. CE and F80 inhibited writhing regarding antinociceptive activity (acetic acid-induced writhing response in mice). In conclusion, CE and F80 have no significant cytotoxic or antitumor activities in cell lines showing low toxicity and no action against tumors in vivo. Both preparations revealed anti-inflammatory and antinociceptive activities, corroborating the pharmacological basis of L. ferrea for ethnomedical use.
RESUMO
OBJECTIVE: The present work reports the purification and partial characterization of an antibacterial lectin (EmaL) obtained from Eugenia malaccensis seeds as well as the evaluation of its effect in the daily topical treatment of repairing process of cutaneous wounds in mice. MATERIALS AND METHODS: The cutaneous wound was produced by the incision of the skin and use of lectin in the treatment of mice cutaneous wounds was evaluated. Surgical wounds were treated daily with a topical administration of EmaL and parameters such as edema, hyperemia, scab, granulation and scar tissues as well as contraction of wounds were analyzed. RESULTS: A novel lectin, with a molecular mass of 14 kDa, was isolated from E. malaccensis using affinity chromatography. The lectin (EmaL) agglutinated glutaraldehyde-treated rabbit and human erythrocytes; the lectin-induced rabbit erythrocyte agglutination was inhibited by glucose, casein, ovalbumin and fetuin. Also, Emal was very effective in the inhibition of bacterial growth, with the best inhibition results obtained for Staphylococcus aureus. Inflammatory signals such as edema and hyperemia were statistically less intense when EmaL was applied compared to the control. The histopathological analysis showed that the treated injured tissue presented reepithelialization (complete or partial) and areas of transition more evidenced than those of the control group, especially due to well organized pattern of collagen fibers presented in the granulation fibrous tissue. CONCLUSION: Presented results are a preliminary indication of the pharmacological interest in using EmaL as antimicrobial agent and in the repairing process of cutaneous wounds.
Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Lectinas/química , Lectinas/farmacologia , Pele/efeitos dos fármacos , Syzygium/química , Cicatrização/efeitos dos fármacos , Administração Cutânea , Animais , Antibacterianos/isolamento & purificação , Eritrócitos/efeitos dos fármacos , Feminino , Tecido de Granulação/efeitos dos fármacos , Humanos , Lectinas/isolamento & purificação , Camundongos , Coelhos , Sementes/química , Staphylococcus aureus/efeitos dos fármacosRESUMO
Cramoll 1,4 is a lectin with specific glucose/mannose binding, which is extracted from seeds of Cratylia mollis Mart. Many assays have shown the cytokine expression activity and anti-inflammatory profile of this lectin. The aim of this study was to evaluate the immunostimulatory response, in vitro, of splenocytes in mice previously inoculated, in vivo, with C. mollis (Cramoll 1,4) and Canavalia ensiformis (Con A) lectins. Results demonstrated higher proliferation indexes induced by Cramoll 1,4 than Con A lectin in relation to all experimental groups. Cramoll 1,4 and Con A also induced high levels of IL-2, IL-6, IFN-γ and nitric oxide production. Moreover, Cramoll 1,4 did not induce apoptosis and stimulated a significant number of cells in the S phase of the cell cycle. Results showed that Cramoll 1,4 lectin induces proliferative response and suggested that this lectin can be used as a mitogenic agent in immunostimulatory assays.
Assuntos
Interferon gama/imunologia , Interleucina-2/imunologia , Interleucina-6/imunologia , Mitose , Lectinas de Plantas/imunologia , Baço/citologia , Baço/imunologia , Animais , Proliferação de Células , Células Cultivadas , Interferon gama/biossíntese , Interleucina-2/biossíntese , Interleucina-6/biossíntese , Linfócitos/citologia , Linfócitos/imunologia , Masculino , Camundongos , Óxido Nítrico/biossínteseRESUMO
AIMS: The in vitro activity of ciclopirox olamine was evaluated against Cryptococcus spp. obtained from the cerebrospinal fluid (CSF) of immunocompromised patients. METHODS AND RESULTS: The antifungal activity of ciclopirox olamine was tested against Cryptococcus spp. obtained from the CSF of immunocompromised patients, using amphotericin B and fluconazole as controls. The minimal inhibitory concentration was determined following the microdilution method indicated by the Clinical and Laboratory Standards Institute. The minimal fungicide concentration was determined by the absence of growth on Sabouraud dextrose agar. The data obtained showed that antifungal activity of ciclopirox olamine ranged from 0·25 to 1 µg ml(-1) . CONCLUSIONS: This paper underscores the importance of the antifungal potential of ciclopirox olamine against Cryptococcus spp. as an alternative treatment against systemic cryptococosis. In vivo experiments are essential for future medical use. SIGNIFICANCE AND IMPACT OF THE STUDY: This was the first time that ciclopirox olamine was tested against Cryptococcus spp. using the reference method. The antifungal activity of this drug against this species suggests an applicable potential for systemic cryptococcosis therapy.
Assuntos
Antifúngicos/farmacologia , Criptococose/microbiologia , Cryptococcus/efeitos dos fármacos , Piridonas/farmacologia , Líquido Cefalorraquidiano/microbiologia , Ciclopirox , Criptococose/tratamento farmacológico , Cryptococcus/crescimento & desenvolvimento , Cryptococcus/isolamento & purificação , Humanos , Testes de Sensibilidade MicrobianaRESUMO
Incubation of T. cruzi epimastigotes with the lectin Cramoll 1,4 in Ca(2+) containing medium led to agglutination and inhibition of cell proliferation. The lectin (50 microg/ml) induced plasma membrane permeabilization followed by Ca(2+) influx and mitochondrial Ca(2+) accumulation, a result that resembles the classical effect of digitonin. Cramoll 1,4 stimulated (five-fold) mitochondrial reactive oxygen species (ROS) production, significantly decreased the electrical mitochondrial membrane potential (Delta Psi(m)) and impaired ADP phosphorylation. The rate of uncoupled respiration in epimastigotes was not affected by Cramoll 1,4 plus Ca(2+) treatment, but oligomycin-induced resting respiration was 65% higher in treated cells than in controls. Experiments using T. cruzi mitochondrial fractions showed that, in contrast to digitonin, the lectin significantly decreased Delta Psi(m) by a mechanism sensitive to EGTA. In agreement with the results showing plasma membrane permeabilization and impairment of oxidative phosphorylation by the lectin, fluorescence microscopy experiments using propidium iodide revealed that Cramoll 1,4 induced epimastigotes death by necrosis.
Assuntos
Fabaceae/química , Lectinas de Plantas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Animais , Sinalização do Cálcio/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Digitonina/farmacologia , Glicoconjugados/metabolismo , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Necrose , Fosforilação Oxidativa/efeitos dos fármacos , Lectinas de Plantas/isolamento & purificação , Espécies Reativas de Oxigênio/metabolismo , Trypanosoma cruzi/citologia , Trypanosoma cruzi/metabolismoRESUMO
AIMS: The aim of this work was to analyse the antimicrobial properties of a purified lectin from Eugenia uniflora L. seeds. METHODS AND RESULTS: The E. uniflora lectin (EuniSL) was isolated from the seed extract and purified by ion-exchange chromatography in DEAE-Sephadex with a purification factor of 11.68. The purified lectin showed a single band on denaturing electrophoresis, with a molecular mass of 67 kDa. EuniSL agglutinated rabbit and human erythrocytes with a higher specificity for rabbit erythrocytes. The haemagglutination was not inhibited by the tested carbohydrates but glycoproteins exerted a strong inhibitory action. The lectin proved to be thermo resistant with the highest stability at pH 6.5 and divalent ions did not affect its activity. EuniSL demonstrated a remarkable nonselective antibacterial activity. EuniSL strongly inhibited the growth of Staphylococcus aureus, Pseudomonas aeruginosa and Klebsiella sp. with a minimum inhibitory concentration (MIC) of 1.5 microg ml(-1), and moderately inhibited the growth of Bacillus subtilis, Streptococcus sp. and Escherichia coli with a MIC of 16.5 microg ml(-1). CONCLUSIONS: EuniSL was found to be effective against bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The strong antibacterial activity of the studied lectin indicates a high potential for clinical microbiology and therapeutic applications.
Assuntos
Antibacterianos/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Lectinas , Sementes/química , Syzygium/química , Hemaglutinação , Testes de Inibição da Hemaglutinação , Humanos , Lectinas/química , Lectinas/isolamento & purificação , Lectinas/metabolismo , Lectinas/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
The electrochemical potentials for free or immobilized Cratylia mollis seed lectin (Cra) were obtained through potentiostatic techniques. A saline solution was used as support to control the charge distribution between saturated calomel electrode and platinum electrode (working electrode). The electrochemical potential to free Cra was determined at the following concentrations: 0.6, 0.9 and 1.0 mg/ml in an aerated environment under different temperatures (5, 10 and 20 degrees C). The best electrochemical potential was obtained with 1.0 mg/ml, at 5 and 10 degrees C, 87 and 102 mV, respectively. Electrochemical potential to Cra immobilized on glass beads activated with 3-aminopropyltriethoxysilane described a linear behavior in relation to the increase in glucose concentration. The development of techniques to define interface electrical parameters will be able to give information about charged groups adsorbed to electrode surface revealing interactions particularly in biological systems.
Assuntos
Eletroquímica , Fabaceae/química , Lectinas/química , Sementes/químicaRESUMO
The effects of mycorrhizal inoculation and increasing soil P levels on the expression of total proteins and peroxidase activity on passion fruit roots were evaluated. The experimental design was entirely at random, with four treatments of inoculation (a - control; b - Gigaspora albida; c - Scutellospora heterogama; d - mixture of G. albida, G. margarita, S. heterogama, and Glomus clarum) Ãù three levels of soil P (4, 11, and 30 mg/dm of soil), each with three replicates. Plants were harvested 70 days after inoculation, when root colonization, shoot P level, protein content, and enzymatic activity of peroxidase (PAGE - 7 percent) on root extract were evaluated. Regarding protein, there was no significant difference among the treatments, except between those roots receiving mixed inoculum and 11 mg P/dm of soil. Effect of P on protein concentration, when compared with the inoculation effect was observed. For peroxidase, there was an eletrophoretic band common to all treatments (rf: 0.43) and another that was absent only in noncolonized plants, grown in soil with lower P (rf: 0.46). Mycorrhizal specific bands were not present but a small decrease of intensity of bands in noncolonized plants was observed. Conversely, the control roots presented a single band (rf: 0.33) not observed in the other extracts, that may demonstrate an inhibitory effect of AMF on some host activities. The data showed the influence of P level in soil on the protein expression of roots, suggesting the influence of this nutrient on root genetic expression as well as on the mechanisms of symbiotic control/recognition
Assuntos
Fungos , Peroxidase , Fósforo , Raízes de Plantas , Proteínas , Solo , Frutas , Fósforo , Raízes de Plantas , Solo , Microbiologia do SoloRESUMO
The effects of mycorrhizal inoculation and increasing soil P levels on the expression of total proteins and peroxidase activity on passion fruit roots were evaluated. The experimental design was entirely at random, with four treatments of inoculation (a--control; b--Gigaspora albida; c--Scutellospora heterogama; d--mixture of G. albida, G. margarita, S. heterogama, and Glomus clarum) x three levels of soil P (4, 11, and 30 mg/dm3 of soil), each with three replicates. Plants were harvested 70 days after inoculation, when root colonization, shoot P level, protein content, and enzymatic activity of peroxidase (PAGE--7%) on root extract were evaluated. Regarding protein, there was no significant difference among the treatments, except between those roots receiving mixed inoculum and 11 mg P/dm3 of soil. Effect of P on protein concentration, when compared with the inoculation effect was observed. For peroxidase, there was an eletrophoretic band common to all treatments (rf: 0.43) and another that was absent only in noncolonized plants, grown in soil with lower P (rf: 0.46). Mycorrhizal specific bands were not present but a small decrease of intensity of bands in noncolonized plants was observed. Conversely, the control roots presented a single band (rf: 0.33) not observed in the other extracts, that may demonstrate an inhibitory effect of AMF on some host activities. The data showed the influence of P level in soil on the protein expression of roots, suggesting the influence of this nutrient on root genetic expression as well as on the mechanisms of symbiotic control/recognition.
Assuntos
Fungos , Passiflora/metabolismo , Peroxidase/metabolismo , Fósforo/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Solo , Passiflora/microbiologia , Fósforo/análise , Raízes de Plantas/química , Raízes de Plantas/microbiologia , Solo/análise , Microbiologia do SoloRESUMO
The phenomenon of altered carbohydrates in transformed cell surfaces has been studied through histochemical techniques using lectins. Specific binding patterns to normal and transformed mammary tissues were evaluated by Isoform 1 from Cratylia mollis lectin (Cra Iso 1). Protocols using a direct method, incubation of Cra Iso I conjugated to peroxidase (Cra Iso 1-Per) with mammary tissues, followed by diaminobenzidine and hydrogen peroxidase interaction, were performed. Neoplastic tissues, marked by Cra Iso 1, showed a higher intensity of staining than normal ones, in comparison with Canavalia ensiformis lectin, Concanavalin A (Con A), conjugated to peroxidase (Con A-Per). The assay with Cra Iso 1 also indicated a possible utilization of this lectin to characterize normal and transformed mammary cells.