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B R Silva, C Maside, L A Vieira, J Cadenas, B G Alves, A C A Ferreira, F L N Aguiar, A L C Silva, J R Figueiredo, J R V Silva
The aim of the present study was to evaluate the effect of frutalin (FTL) on in vitro maturation (IVM), and fertilization (IVF) of pig oocytes. In the Experiment 1, cumulus-oocyte complexes (COCs) were submitted to IVM in maturation medium alone or supplemented with different FTL concentration (0.6, 6 and 60 μg/mL), or 0.3 μg/mL doxorubicin (DXR). After IVM, some oocytes were evaluated for chromatin configuration, and the remaining oocytes were submitted to in vitro fertilization. In Experiment 2, matured oocytes were fertilized in IVF medium alone (control) or in presence of different FTL concentration (0...
May 2018: Animal Reproduction Science
João Ronielly Campêlo Araújo, José de Maria Albuquerque de Melo Júnior, Marina de Barros Mamede Vidal Damasceno, Sacha Aubrey Alves Rodrigues Santos, Antônio Eufrásio Vieira-Neto, Marina Duarte Pinto Lobo, Adriana Rolim Campos, Renato de Azevedo Moreira, Ana Cristina de Oliveira Monteiro-Moreira
In this study we evaluated the effect of frutalin (FTL) on mouse behavior. Mice (n=6/group) were treated (i.p.) with FTL (0.25; 0.5 or 1mg/kg) or vehicle and submitted to several tests (hole-board/HBT, elevated plus maze/PMT, open field/OFT, tail suspension/TST, or forced swimming/FST). Yohimbine, ketamine, l-NAME, aminoguanidine, 7-NI, methylene blue, l-arginine or dl-serine was administered 30min before FTL (0.5mg/kg). To evaluate the subchronic effect, animals were injected with FTL or vehicle for 7days and submitted to the FST...
June 2018: International Journal of Biological Macromolecules
Felipe Domingos de Sousa, Bruno Bezerra da Silva, Gilvan Pessoa Furtado, Igor de Sa Carneiro, Marina Duarte Pinto Lobo, Yiwei Guan, Jingxu Guo, Alun R Coker, Marcos Roberto Lourenzoni, Maria Izabel Florindo Guedes, James S Owen, David J Abraham, Ana Cristina de Oliveira Monteiro-Moreira, Renato de Azevedo Moreira
Artocarpus incisa (breadfruit) seeds contain three different lectins (Frutalin, Frutapin (FTP) and Frutackin) with distinct carbohydrate specificities. The most abundant lectin is Frutalin, an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and potential for tumour biomarker discovery as already reported. FTP is the second most abundant, but proved difficult to purify with very low yields and contamination with Frutalin frustrating its characterization. Here, we report for the first time high-level production and isolation of biologically active recombinant FTP in Escherichia coli BL21, optimizing conditions with the best set yielding >40 mg/l culture of soluble active FTP...
August 31, 2017: Bioscience Reports
Marina Duarte Pinto Lobo, Frederico Bruno Mendes Batista Moreno, Gustavo Henrique Martins Ferreira Souza, Sara Maria Moreira Lima Verde, Renato de Azevedo Moreira, Ana Cristina de Oliveira Monteiro-Moreira
Breast cancer is one of the most commonly diagnosed types of cancer among women. Breast cancer mortality rates remain high probably because its diagnosis is hampered by inaccurate detection methods. Since changes in protein expression as well as modifications in protein glycosylation have been frequently reported in cancer development, the aim of this work was to study the differential expression as well as modifications of glycosylation of proteins from plasma of women with breast cancer at different stages of disease (n = 30) compared to healthy women (n = 10)...
2017: Frontiers in Oncology
Maria A A Soares, José J N Costa, Gisvani L Vasconcelos, Regislane P Ribeiro, José C Souza, André L C Silva, Robert Van den Hurk, José R V Silva
Frutalin is a galactose-binding lectin that has an irreversible cytotoxic effect on HeLa cervical cancer cells, by inducing apoptosis and inhibiting cell proliferation. It was previously shown that after in vitro incubation, frutalin is internalized into HeLa cells nucleus, which indicates that frutalin apoptosis-inducing activity might be linked with its nuclear localization. Considering that drugs commonly used for cancer treatment have a deleterious effect on germ cells, the aim of this study was to evaluate the effect of frutalin on the activation, survival, ultrastructure and gene expression in follicles cultured within ovarian tissue...
January 2018: Histology and Histopathology
Marina B M V Damasceno, José de Maria A de Melo Júnior, Sacha Aubrey A R Santos, Luana T M Melo, Laura Hévila I Leite, Antonio E Vieira-Neto, Renato de A Moreira, Ana Cristina de O Monteiro-Moreira, Adriana R Campos
Orofacial pain is a highly prevalent clinical condition, yet difficult to control effectively with available drugs. Much attention is currently focused on the anti-inflammatory and antinociceptive properties of lectins. The purpose of this study was to evaluate the antinociceptive effect of frutalin (FTL) using rodent models of inflammatory and neuropathic orofacial pain. Acute pain was induced by formalin, glutamate or capsaicin (orofacial model) and hypertonic saline (corneal model). In one experiment, animals were pretreated with l-NAME and naloxone to investigate the mechanism of antinociception...
August 25, 2016: Chemico-biological Interactions
Ana Cristina de Oliveira Monteiro-Moreira, Humberto D'Muniz Pereira, Antonio Eufrasio Vieira Neto, Frederico Bruno Mendes Batista Moreno, Marina Duarte Pinto Lobo, Felipe Domingos de Sousa, Renato de Azevedo Moreira
Frutalin is an α-D-galactose-specific carbohydrate-binding glycoprotein with antitumour properties and is a powerful tool for tumour biomarker discovery. The crystallization and preliminary X-ray diffraction analysis of this lectin, which was isolated from Artocarpus incisa seeds, are reported here. Frutalin was purified and submitted to mass-spectrometric analysis. Diverse masses at approximately 16 kDa were observed in the deconvoluted spectra, which support the presence of isoforms. The best frutalin crystals were grown within a week in 0...
October 2015: Acta Crystallographica. Section F, Structural Biology Communications
Carla Oliveira, José A Teixeira, Lucília Domingues
Frutalin is a homotetrameric partly glycosylated α-D-galactose-binding lectin of biomedical interest from Artocarpus incisa (breadfruit) seeds, belonging to the jacalin-related lectins family. As other plant lectins, frutalin is a heterogeneous mixture of several isoforms possibly with distinct biological activities. The main problem of using such lectins as biomedical tools is that "batch-to-batch" variation in isoforms content may lead to inconstant results. The production of lectins by recombinant means has the advantage of obtaining high amounts of proteins with defined amino-acid sequences and more precise properties...
2014: Frontiers in Plant Science
Marlene Lopes, Carla Oliveira, Lucília Domingues, Manuel Mota, Isabel Belo
Pichia pastoris is a widely used host for the production of heterologous proteins. In this case, high cell densities are needed and oxygen is a major limiting factor. The increased air pressure could be used to improve the oxygen solubility in the medium and to reach the high oxygen demand of methanol metabolism. In this study, two P. pastoris strains producing two different recombinant proteins, one intracellular (β-galactosidase) and other extracellular (frutalin), were used to investigate the effect of increased air pressure on yeast growth in glycerol and heterologous protein production, using the methanol AOX1-inducible system...
September 2014: Biotechnology Progress
Sofia J Costa, André Almeida, António Castro, Lucília Domingues, Hüseyin Besir
The Escherichia coli host system is an advantageous choice for simple and inexpensive recombinant protein production but it still presents bottlenecks at expressing soluble proteins from other organisms. Several efforts have been taken to overcome E. coli limitations, including the use of fusion partners that improve protein expression and solubility. New fusion technologies are emerging to complement the traditional solutions. This work evaluates two novel fusion partners, the Fh8 tag (8 kDa) and the H tag (1 kDa), as solubility enhancing tags in E...
August 2013: Applied Microbiology and Biotechnology
Ana Paula de Vasconcellos Abdon, Greicy Coelho de Souza, Lílian Noronha Coelho de Souza, Renata Prado Vasconcelos, Carolina Araújo Castro, Marjorie Moreira Guedes, Roberto César Pereira Lima, Renato de Azevedo Moreira, Ana Cristina de Oliveira Monteiro-Moreira, Adriana Rolim Campos
The present study was designed to verify whether frutalin (FTL) affords gastroprotection against the ethanol-induced gastric damage and to examine the underlying mechanism(s). Gastric damage was induced by intragastric administration of 0.2 ml of ethanol (96%). Mice in groups were pretreated with FTL (0.25, 0.5 and 1 mg/kg; i.p.), cimetidine (100 mg/kg; p.o.), or vehicle (0.9% of NaCl, 10 mL/kg; p.o.), 30 min before ethanol administration. They were sacrificed 30 min later, the stomachs excised, and the mucosal lesion area (mm²) measured by planimetry...
April 2012: Fitoterapia
Carla Oliveira, Ana Nicolau, José A Teixeira, Lucília Domingues
Frutalin is the α-D-galactose-binding lectin isolated from breadfruit seeds. Frutalin was obtained from two different sources: native frutalin was purified from its natural origin, and recombinant frutalin was produced and purified from Pichia pastoris. This work aimed to study and compare the effect of native and recombinant frutalin on HeLa cervical cancer cells proliferation and apoptosis. Furthermore, the interaction between frutalin and the HeLa cells was investigated by confocal microscopy. Despite having different carbohydrate-binding affinities, native and recombinant frutalin showed an identical magnitude of cytotoxicity on HeLa cells growth (IC₅₀~100 μg/mL) and equally induced cell apoptosis...
2011: Journal of Biomedicine & Biotechnology
Thatyane M Nobre, Felippe J Pavinatto, Márcia R Cominetti, Heloísa S Selistre de-Araújo, Maria E D Zaniquelli, Leila M Beltramini
Frutalin is a homotetrameric alpha-d-galactose (d-Gal)-binding lectin that activates natural killer cells in vitro and promotes leukocyte migration in vivo. Because lectins are potent lymphocyte stimulators, understanding the interactions that occur between them and cell surfaces can help to the action mechanisms involved in this process. In this paper, we present a detailed investigation of the interactions of frutalin with phospho- and glycolipids using Langmuir monolayers as biomembrane models. The results confirm the specificity of frutalin for d-Gal attached to a biomembrane...
August 2010: Biochimica et Biophysica Acta
Carla Oliveira, José A Teixeira, Fernando Schmitt, Lucília Domingues
BACKGROUND: Numerous studies indicate that cancer cells present an aberrant glycosylation pattern that can be detected by lectin histochemistry. Lectins have shown the ability to recognise these modifications in several carcinomas, namely in the prostate carcinoma, one of the most lethal diseases in man. Thus, the aim of this work was to investigate if the alpha-D-galactose-binding plant lectin frutalin is able to detect such changes in the referred carcinoma. Frutalin was obtained from different sources namely, its natural source (plant origin) and a recombinant source (Pichia expression system)...
2009: BMC Biotechnology
Carla Oliveira, Sofia Costa, José A Teixeira, Lucília Domingues
cDNA clones encoding frutalin, the alpha-D-galactose-binding lectin expressed in breadfruit seeds (Artocarpus incisa), were isolated and sequenced. The deduced amino acid sequences indicated that frutalin may be encoded by a family of genes. The NCBI database searches revealed that the frutalin sequence is highly homologous with jacalin and mornigaG sequences. Frutalin cDNA was re-amplified and cloned into the commercial expression vector pET-25b(+) for frutalin production in Escherichia coli. An experimental factorial design was employed to maximise the soluble expression of the recombinant lectin...
November 2009: Molecular Biotechnology
Carla Oliveira, Wagner Felix, Renato A Moreira, José A Teixeira, Lucília Domingues
Frutalin is an alpha-D-galactose-binding lectin expressed in breadfruit seeds. Its isolation from plant is time-consuming and results in a heterogeneous mixture of different lectin isoforms. In order to improve and facilitate the availability of the breadfruit lectin, we cloned an optimised codifying frutalin mature sequence into the pPICZalphaA expression vector. This expression vector, designed for protein expression in the methylotrophic yeast Pichia pastoris, contains the Saccharomyces alpha-factor preprosequence to direct recombinant proteins into the secretory pathway...
August 2008: Protein Expression and Purification
S G De-Simone, C C Netto, F P Silva
Affinity chromatography based on the commercial resin Sepharose CL-6B was used to isolate new C1-beta-type lectins from crude preparations of snake venoms (Bothrops jararaca, Bothrops jararacussu, Bothrops newiedi, Bothrops moojeni, Lachesis muta rhombeata). Most of the C-type lectins could be eluted with almost 100% recovery using the competitor isopropyl-beta-D-thiogalactoside (IPTG) or through Ca2+ sequestration with EDTA. The lectin yield varied considerably among the different snake species, but B. newiedi venom was a particularly rich source of lectin, retaining 2...
July 11, 2006: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Aline C Brando-Lima, Roberta F Saldanha-Gama, Cristiane Ribeiro Pereira, Christina Gaspar Villela, André Luiz Franco Sampaio, Ana C O Monteiro-Moreira, Maria das Graças M O Henriques, Renato A Moreira, Christina Barja-Fidalgo
The mechanisms involved in the mitogenic effect of lectins are not fully understood and are thought to involve a cascade of intracellular signals related to T cell receptor activation. This study shows that frutalin, the alpha-D-galactose-binding lectin from Artocarpus incisa seeds, is a potent mitogenic activator of human lymphocytes. This effect is inhibited by D-galactose and PI3K inhibitors, and is accompanied by an increase in IL-2 receptor expression and by a PI3K-dependent IL-2 gene expression and IL-2 protein synthesis...
March 2006: International Immunopharmacology
Aline C Brando-Lima, Roberta F Saldanha-Gama, Maria das Graças M O Henriques, Ana C O Monteiro-Moreira, Renato A Moreira, Christina Barja-Fidalgo
Several lectin-like molecules have been shown as potent activators of leukocytes. Galactose-binding lectins are of special interest since they could interact with several endogenous molecules involved in the innate and specific immune responses. The effects of Frutalin (FTL), an alpha-D-galactose (Gal)-binding plant lectin, on the modulation of neutrophil (PMN) functions were investigated. FTL induced a dose-dependent PMN migration in mice pleural cavity. Moreover, FTL was also a potent direct chemotactic for human PMN, in vitro, and triggered oxidative burst in these cells...
October 15, 2005: Toxicology and Applied Pharmacology
Patricia T Campana, Derminda I Moraes, Ana C O Monteiro-Moreira, Leila M Beltramini
Protein refolding is currently a fundamental problem in biophysics and molecular biology. We have studied the refolding process of frutalin, a tetrameric lectin that presents structural homology with jacalin but shows a more marked biological activity. The initial state in our refolding puzzle was that proteins were unfolded after thermal denaturation or denaturation induced by guanidine hydrochloride, and under both conditions, frutalin was refolded. The denaturation curves, measured by fluorescence emission, gave values of conformational stability of 17...
February 2002: European Journal of Biochemistry
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