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1.
Anticancer Drugs ; 34(10): 1085-1093, 2023 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-37823283

RESUMO

Every food source contains both edible and inedible waste components. Millions of tonnes of trash from the food business are made from fruits, and these wastes are containing higher-value medicinal components, such as alkaloids, flavonoids, phenolic contents, a huge amount of proteins and secondary metabolites. These bioactive phytoconstituents are being used for the treatment of many serious fatal diseases. So, utilizing the recovered bioactive molecules from food wastes as functional ingredients offers a long-term alternative source of therapeutically active components that will lead to the discovery of novel phytoconstituents or novel treatment approaches. The goal of this systematic study is to provide an overview of the jackfruit (Artocarpus heterophyllus Lam, Moraceae) edible byproducts, such as jackfruit seeds that are largely neglected. This seed contains numerous bioactive lead molecules, such as carbohydrate-binding protein jacalin, which exhibits potent anticancer activity against colon cancer, blood cancer and breast cancer as well as can enlighten the new possible treatment approaches in targeted therapy and photodynamic chemotherapy. Moreover, jackfruit waste seed can be taken as a dietary food, which is having property to prevent and treat cancer and other lifestyle diseases. The works that have been carried out to utilize jackfruit waste other than the juicy edible bulbs have been reviewed in this article.


Assuntos
Artocarpus , Lectinas , Humanos , Lectinas/análise , Lectinas/química , Artocarpus/química , Lectinas de Plantas/análise , Sementes/química
2.
J Int AIDS Soc ; 23(10): e25628, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33073530

RESUMO

INTRODUCTION: The majority of new HIV infections occur through mucosal transmission. The availability of readily applicable and accessible platforms for anti-retroviral (ARV) delivery is critical for the prevention of HIV acquisition through sexual transmission in both women and men. There is a compelling need for developing new topical delivery systems that have advantages over the pills, gels and rings, which currently fail to guarantee protection against mucosal viral transmission in vulnerable populations due to lack of user compliance. The silk fibroin (SF) platform offers another option that may be better suited to individual circumstances and preferences to increase efficacy through user compliance. The objective of this study was to test safety and efficacy of SF for anti-HIV drug delivery to mucosal sites and for viral prevention. METHODS: We formulated a potent HIV inhibitor Griffithsin (Grft) in a mucoadhesive silk fibroin (SF) drug delivery platform and tested the application in a non-human primate model in vivo and a pre-clinical human cervical and colorectal tissue explant model. Both vaginal and rectal compartments were assessed in rhesus macaques (Mucaca mulatta) that received SF (n = 4), no SF (n = 7) and SF-Grft (n = 11). In this study, we evaluated the composition of local microbiota, inflammatory cytokine production, histopathological changes in the vaginal and rectal compartments and mucosal protection after ex vivo SHIV challenge. RESULTS: Effective Grft release and retention in mucosal tissues from the SF-Grft platform resulted in protection against HIV in human cervical and colorectal tissue as well as against SHIV challenge in both rhesus macaque vaginal and rectal tissues. Mucoadhesion of SF-Grft inserts did not cause any inflammatory responses or changes in local microbiota. CONCLUSIONS: We demonstrated that in vivo delivery of SF-Grft in rhesus macaques fully protects against SHIV challenge ex vivo after two hours of application and is safe to use in both the vaginal and rectal compartments. Our study provides support for the development of silk fibroin as a highly promising, user-friendly HIV prevention modality to address the global disparity in HIV infection.


Assuntos
Fármacos Anti-HIV/administração & dosagem , Fibroínas , Infecções por HIV/prevenção & controle , Lectinas/administração & dosagem , Lectinas de Plantas/administração & dosagem , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Animais , Fármacos Anti-HIV/análise , Fármacos Anti-HIV/farmacocinética , Materiais Biocompatíveis , Colo do Útero/virologia , Colo/virologia , Feminino , Microbioma Gastrointestinal/efeitos dos fármacos , HIV/efeitos dos fármacos , Humanos , Lectinas/análise , Lectinas/farmacocinética , Macaca mulatta , Microbiota/efeitos dos fármacos , Mucosa/química , Veículos Farmacêuticos , Lectinas de Plantas/análise , Lectinas de Plantas/farmacocinética , Reto/química , Reto/microbiologia , Reto/virologia , Vagina/química , Vagina/microbiologia
3.
J Cell Mol Med ; 24(18): 11012-11017, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32744427

RESUMO

Traumatic nerve injuries have become a common clinical problem, and axon regeneration is a critical process in the successful functional recovery of the injured nervous system. In this study, we found that peripheral axotomy reduces PTEN expression in adult sensory neurons; however, it did not alter the expression level of PTEN in IB4-positive sensory neurons. Additionally, our results indicate that the artificial inhibition of PTEN markedly promotes adult sensory axon regeneration, including IB4-positive neuronal axon growth. Thus, our results provide strong evidence that PTEN is a prominent repressor of adult sensory axon regeneration, especially in IB4-positive neurons.


Assuntos
Regeneração Nervosa/fisiologia , Proteínas do Tecido Nervoso/antagonistas & inibidores , Crescimento Neuronal/fisiologia , PTEN Fosfo-Hidrolase/antagonistas & inibidores , Fenantrenos/farmacologia , Lectinas de Plantas/análise , Neuropatia Ciática/fisiopatologia , Células Receptoras Sensoriais/metabolismo , Animais , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Gânglios Espinais/citologia , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Camundongos Knockout , Regeneração Nervosa/efeitos dos fármacos , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/deficiência , Proteínas do Tecido Nervoso/genética , Crescimento Neuronal/efeitos dos fármacos , PTEN Fosfo-Hidrolase/deficiência , PTEN Fosfo-Hidrolase/fisiologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Células Receptoras Sensoriais/química , Células Receptoras Sensoriais/classificação , Células Receptoras Sensoriais/efeitos dos fármacos
4.
Anal Bioanal Chem ; 411(23): 6049-6056, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31280477

RESUMO

As an important glycoprotein of the lectin family, soybean agglutinin (SBA) is an anti-nutritional factor with considerable toxic and side effects and plays a significant role in tumor analysis. In order to achieve the sensitive detection of SBA, a sandwich-structured electrochemiluminescence (ECL) biosensor was constructed using carboxylated carbon nitride (C-g-C3N4) as luminophore and D-galactosamine (galM) as a recognition element. A glassy carbon electrode (GCE) was modified with Au nanoparticles (Au NPs) for capturing the galM via Au-N bond, and further capturing the target SBA by specific recognition between galM and SBA. In the presence of SBA, the composite C-g-C3N4-galM was immobilized onto the electrode. With the increase in the concentration of SBA, the ECL signal from C-g-C3N4 increased, thus achieving a signal-on detection of SBA. The linear range of the biosensor was 1.0 ng/mL~10 µg/mL and detection limit for SBA was as low as 0.33 ng/mL. In this construction strategy, C-g-C3N4 not only acted as an excellent signal probe, but also as an immobilization matrix to easily achieve a high loading of the small molecule recognition element galM. This strategy provides a simple alternative SBA detection platform. Graphical abstract.


Assuntos
Galactosamina/química , Grafite/química , Substâncias Luminescentes/química , Nitrilas/química , Lectinas de Plantas/análise , Proteínas de Soja/análise , Técnicas Biossensoriais/métodos , Ácidos Carboxílicos/química , Técnicas Eletroquímicas/métodos , Ouro/química , Humanos , Limite de Detecção , Medições Luminescentes/métodos , Nanopartículas Metálicas/química , Lectinas de Plantas/sangue , Proteínas de Soja/sangue
5.
J Neuroimmunol ; 332: 198-211, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31077855

RESUMO

Excitation of dorsal root ganglion (DRG) neurons by interleukin 1ß (IL-1ß) is implicated in the onset of neuropathic pain. To understand its mechanism of action, isolectin B4 positive (IB4+) DRG neurons were exposed to 100pM IL-1ß for 5-6d. A reversible increase in action potential (AP) amplitude reflected increased TTX-sensitive sodium current (TTX-S INa). An irreversible increase in AP duration reflected decreased Ca2+- sensitive K+ conductance (BK(Ca) channels). Different processes thus underlie regulation of the two channel types. Since changes in AP shape facilitated Ca2+ influx, this explains how IL-1ß facilitates synaptic transmission in the dorsal horn; thereby provoking pain.


Assuntos
Canais de Cálcio/efeitos dos fármacos , Gânglios Espinais/citologia , Interleucina-1beta/farmacologia , Ativação do Canal Iônico/efeitos dos fármacos , Neuralgia/etiologia , Canais de Potássio/efeitos dos fármacos , Células Receptoras Sensoriais/efeitos dos fármacos , Canais de Sódio/efeitos dos fármacos , Potenciais de Ação/efeitos dos fármacos , Animais , Canais de Cálcio/metabolismo , Tamanho Celular , Fator Neurotrófico Derivado de Linhagem de Célula Glial/farmacologia , Masculino , Fator de Crescimento Neural/farmacologia , Neuralgia/metabolismo , Técnicas de Patch-Clamp , Peptídeos/farmacologia , Lectinas de Plantas/análise , Canais de Potássio/metabolismo , Ratos , Ratos Sprague-Dawley , Células Receptoras Sensoriais/metabolismo , Células Receptoras Sensoriais/ultraestrutura , Canais de Sódio/metabolismo
6.
J Gastroenterol ; 54(6): 549-557, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30707282

RESUMO

BACKGROUND: Nonalcoholic steatohepatitis (NASH) is associated with liver inflammation in patients with nonalcoholic fatty liver disease, and it can progress to liver fibrosis at an advanced stage, as well as hepatocellular carcinoma (HCC) and portal hypertension. Although liver fibrosis is accurately diagnosed via biopsy, noninvasive methods are preferable. Aldo-keto reductase family 1 member B10 (AKR1B10) is associated with HCC and is secreted into the blood by liver cells via a lysosome-mediated nonclassical pathway. Accordingly, we analyzed whether secretion of AKR1B10 protein is associated with advanced NASH. METHODS: We performed histological staging in 85 Matteoni classification type III and IV NASH patients and evaluated the incidence of HCC, formation of gastroesophageal varices, and prognosis according to serum AKR1B10 and Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA(+)-M2BP)(M2BPGi) and by comparison with conventional markers of fibrosis. RESULTS:  A positive correlation was found between the Brunt classification and serum AKR1B10 level. In Brunt stage 4 patients, AKR1B10 levels were higher than those of other liver fibrosis markers, with higher specificity. The cutoff values for AKR1B10 and WFA(+)-M2BP for stage 4 fibrosis were 1.03 and 3.11, respectively. The rates of stage 4 fibrosis, HCC incidence, and gastroesophageal varix formation were significantly different between the two groups subdivided according to these cutoff levels. Moreover, the patients in the higher value group had significantly worse prognosis after NASH diagnosis CONCLUSION: AKR1B10 is a useful serum biomarker for advanced liver fibrosis in NASH and, combined with serum WFA(+)-M2BP, can predict HCC development, gastroesophageal varix formation, and poor prognosis.


Assuntos
Aldo-Ceto Redutases/sangue , Carcinoma Hepatocelular/epidemiologia , Cirrose Hepática/patologia , Neoplasias Hepáticas/epidemiologia , Hepatopatia Gordurosa não Alcoólica/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/patologia , Progressão da Doença , Feminino , Humanos , Cirrose Hepática/sangue , Cirrose Hepática/etiologia , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/sangue , Hepatopatia Gordurosa não Alcoólica/fisiopatologia , Lectinas de Plantas/análise , Prognóstico , Receptores de N-Acetilglucosamina/análise
7.
Ann Lab Med ; 38(4): 348-354, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29611385

RESUMO

BACKGROUND: Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA-M2BP) is a protein with altered glycosylation that reacts with lectin, and was recently identified as a useful non-invasive biomarker for the diagnosis of liver fibrosis in patients with hepatitis C virus infection.This study aimed to evaluate the diagnostic efficacy of WFA-M2BP for liver fibrosis in the context of hepatitis B virus (HBV). METHODS: We enrolled 151 patients infected with HBV. Liver biopsy and elastography (Fibroscan) were performed during the initial visit. Fibrosis was graded according to the Knodell histologic activity index (F0-3). WFA-M2BP levels were determined with an automated immunoassay analyzer (M2BPGi, HISCL-5000, Sysmex, Japan). The diagnostic efficacy of WFA-M2BP was compared with those of various conventional or composite biomarkers, including enhanced liver fibrosis (ELF) score, Fibroscan, aspartate transaminase (AST)-to-platelet ratio index (APRI), and FIB-4, based on the area under the ROC curve (AUC) value. RESULTS: The majority of patients were at fibrosis stages F1 and F2. The F2 and F3 AUC values for WFA-M2BP were similar to those for FIB-4, APRI, ELF, and Fibroscan, although the latter showed the best diagnostic efficacy. The diagnostic accuracy of all tested biomarkers for F2 and F3 was 60-70%. In multivariate analysis, WFA-M2BP, ELF, and platelet count significantly predicted stage ≥F2, whereas only platelet count significantly predicted F3. CONCLUSIONS: WFA-M2BP can support a diagnosis of liver fibrosis with similar diagnostic efficacy to other biomarkers, and predicted liver fibrosis stage ≥2 among patients with chronic hepatitis B.


Assuntos
Antígenos de Neoplasias/análise , Hepatite B Crônica/complicações , Imunoensaio , Cirrose Hepática/diagnóstico , Glicoproteínas de Membrana/análise , Lectinas de Plantas/análise , Receptores de N-Acetilglucosamina/análise , Adulto , Área Sob a Curva , Aspartato Aminotransferases/análise , Biomarcadores/análise , Plaquetas/citologia , Técnicas de Imagem por Elasticidade , Feminino , Hepatite B Crônica/diagnóstico , Humanos , Fígado/diagnóstico por imagem , Fígado/patologia , Cirrose Hepática/complicações , Cirrose Hepática/patologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Contagem de Plaquetas , Curva ROC , Estudos Retrospectivos
8.
Anal Chim Acta ; 1001: 158-167, 2018 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-29291799

RESUMO

For use in the voltammetric sensing of galactose-dependent proteins, we modified magnetic beads with a peptide that had both electroactive- and molecular recognition properties. The peptide consisted of a YXY sequence and behaved as an electron-transfer carbohydrate-mimetic peptide that would combine with proteins. With this tool, the protein could be detected via a label-free system. We synthesized several penta- and hexa-peptides with a cysteine residue on the C-terminals to examine the properties of peptides. These peptides contained amino acid residues (X) of alanine, serine, or tyrosine. The peptides were immobilized on magnetic beads via N-(8-maleimidocapryloxy) succinimide. Soybean agglutinin(SBA), the in vivo function of which has been well established in animals, was selected as a model protein. The protein was detected via the changes in electrode response due to the oxidation of tyrosine residues from the phenol group to quinone. As a result, SBA was selectively accumulated on the beads modified with YYYYC. The calibration curve of SBA was linear and ranged from 2.5 × 10-12 to 1.0 × 10-10 M. With this system, SBA was recovered in human serum at values that ranged from 98 to 103%. Furthermore, the beads with peptides were regenerated five times using a protein denaturant. Accordingly, this electrochemical system was simple and could be rapidly applied to the detection of galactose-recognition proteins.


Assuntos
Técnicas Biossensoriais/métodos , Imãs/química , Peptídeos/química , Lectinas de Plantas/análise , Proteínas de Soja/análise , Sequência de Aminoácidos , Técnicas Biossensoriais/instrumentação , Transporte de Elétrons , Desenho de Equipamento , Galactose/metabolismo , Humanos , Masculino , Peptídeos/metabolismo , Lectinas de Plantas/sangue , Lectinas de Plantas/metabolismo , Proteínas de Soja/sangue , Proteínas de Soja/metabolismo , Glycine max/química , Glycine max/metabolismo
9.
J Hepatol ; 67(5): 933-939, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28627363

RESUMO

BACKGROUND AND AIMS: Although treatment for hepatitis C virus has been dramatically improved by the development of direct-acting antiviral agents (DAAs), whether interferon (IFN)-free therapy reduces hepatocarcinogenesis in an equivalent manner to IFN-based therapy remains controversial. The aims of this study were to evaluate the occurrence and recurrence of hepatocellular carcinoma (HCC) in chronic hepatitis C (CHC) patients treated with DAAs and to identify biomarkers of HCC development after antiviral treatment. METHODS: A restrospective review of a prospective database of 1,897 CHC patients who were treated with IFN-based (1,145) or IFN-free therapies (752) was carried out. Cumulative HCC occurrence and recurrence rates were compared using propensity score-matched analysis. Predictors of HCC development after viral eradication were identified by multivariate analysis. RESULTS: Propensity score-matched analysis showed no significant difference in HCC occurrence (p=0.49) and recurrence rates (p=0.54) between groups treated with IFN-based or IFN-free therapies. In multivariate analysis, higher levels of post-treatment α-fetoprotein (AFP) or Wisteria floribunda agglutinin positive Mac-2 binding protein (WFA+M2BP) were independently associated with HCC occurrence and recurrence after viral eradication. Only post-treatment WFA+M2BP level was significantly associated with HCC occurrence and recurrence among patients without severe fibrosis. The area under the receiver operating characteristic (ROC) curve for WFA+M2BP levels was greater than that for AFP levels in ROC analysis. CONCLUSION: The risks of early HCC occurrence and recurrence after viral eradication were similar between IFN-based and IFN-free therapies. Post-treatment levels of WFA+M2BP may be helpful screening biomarkers for assessing the risk of HCC after IFN-free therapy. Patients with high WFA+M2BP levels after antiviral treatment, even without severe fibrosis, must be followed up carefully for HCC development. Lay summary: The risks of early HCC occurrence and recurrence after viral eradication were similar between IFN-based and IFN-free therapies. Post-treatment levels of WFA+M2BP may be helpful screening biomarkers for assessing the risk of HCC after IFN-free therapy.


Assuntos
Antivirais/administração & dosagem , Carcinoma Hepatocelular/prevenção & controle , Hepatite C Crônica , Interferons/administração & dosagem , Neoplasias Hepáticas/prevenção & controle , Recidiva Local de Neoplasia , Adulto , Idoso , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/fisiopatologia , Feminino , Hepatite C Crônica/complicações , Hepatite C Crônica/tratamento farmacológico , Humanos , Japão/epidemiologia , Neoplasias Hepáticas/etiologia , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/fisiopatologia , Glicoproteínas de Membrana/análise , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/etiologia , Recidiva Local de Neoplasia/virologia , Lectinas de Plantas/análise , Receptores de N-Acetilglucosamina/análise , Medição de Risco/métodos , Fatores de Risco , alfa-Fetoproteínas/análise
10.
J Hematol Oncol ; 10(1): 104, 2017 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-28494777

RESUMO

BACKGROUND: Studies in murine models suggested that platelet desialylation was an important mechanism of thrombocytopenia during sepsis. METHODS: First, we performed a prospective, multicenter, observational study that enrolled septic patients with or without thrombocytopenia to determine the association between platelet desialylation and thrombocytopenia in patients with sepsis, severe sepsis, and septic shock. Gender- and age-matched healthy adults were selected as normal controls in analysis of the platelet desialylation levels (study I). Next, we conducted an open-label randomized controlled trial (RCT) in which the patients who had severe sepsis with thrombocytopenia (platelet counts ≤50 × 109/L) were randomly assigned to receive antimicrobial therapy alone (control group) or antimicrobial therapy plus oseltamivir (oseltamivir group) in a 1:1 ratio (study II). The primary outcomes were platelet desialylation level at study entry, overall platelet response rate within 14 days post-randomization, and all-cause mortality within 28 days post-randomization. Secondary outcomes included platelet recovery time, the occurrence of bleeding events, and the amount of platelets transfused within 14 days post-randomization. RESULTS: The platelet desialylation levels increased significantly in the 127 septic patients with thrombocytopenia compared to the 134 patients without thrombocytopenia. A platelet response was achieved in 45 of the 54 patients in the oseltamivir group (83.3%) compared with 34 of the 52 patients in the control group (65.4%; P = 0.045). The median platelet recovery time was 5 days (interquartile range 4-6) in the oseltamivir group compared with 7 days (interquartile range 5-10) in the control group (P = 0.003). The amount of platelets transfused decreased significantly in the oseltamivir group compared to the control group (P = 0.044). There was no difference in the overall 28-day mortality regardless of whether oseltamivir was used. The Sequential Organ Failure Assessment score and platelet recovery time were independent indicators of oseltamivir therapy. The main reason for all of the mortalities was multiple-organ failure. CONCLUSIONS: Thrombocytopenia was associated with increased platelet desialylation in septic patients. The addition of oseltamivir could significantly increase the platelet response rate, shorten platelet recovery time, and reduce platelet transfusion. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR-IPR-16008542 .


Assuntos
Plaquetas/química , Ácido N-Acetilneuramínico/sangue , Sepse/complicações , Trombocitopenia/terapia , Adulto , Especificidade de Anticorpos , Receptor de Asialoglicoproteína/fisiologia , Autoanticorpos/imunologia , Biomarcadores , Monitoramento de Medicamentos/métodos , Feminino , Citometria de Fluxo , Humanos , Fragmentos Fc das Imunoglobulinas/imunologia , Pessoa de Meia-Idade , Lectinas de Plantas/análise , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/imunologia , Complexo Glicoproteico GPIb-IX de Plaquetas/imunologia , Púrpura Trombocitopênica Idiopática/terapia , Trombocitopenia/etiologia , Resultado do Tratamento
11.
J Mol Recognit ; 30(2)2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27774692

RESUMO

Three isolectins denoted hereforth MBaL-30, MBaL-60, and MBaL-80 were isolated from seeds extract of Momordica balsamina by 30%, 60%, and 80% ammonium sulfate saturations, respectively. The native molecular weights of these lectins, as judged by gel filtration, were 108, 56, and 160 kDa, respectively. On SDS-PAGE, under reduced condition, 27 kDa band was obtained for all isolectins. The lectins hemagglutinating activities were variably inhibited by d-galactose (minimum inhibitory concentrations = 12.5mM, 50mM, and 0.391mM, respectively). MBaL-30 and -60 could agglutinate all human blood types with slight preference for the A and O blood groups, whereas MBaL-80 did not agglutinate B and AB blood types. The 3 isolectins were purified from crude seeds extract, collectively, in a single step on the affinity matrix Lactamyl-Seralose 4B; this purified lectin fraction, which contains all isolectins, is termed MBaL. The N-terminal of MBaL till the 25th amino acid was NLSLSELDFSADTYKSFIKNLRKQL, which shares 88% sequence identity with Momordica charantia lectin type-2 ribosomal inactivating protein from Momordica charantia and 50% with momordin II from Momordica balsamina. MBaL retained 100% activity at up to 50°C for 30 minutes. MBaL-30 and MBaL-60 exhibited maximum activities in the pH range between 4 and 8, while MBaL-80 was showing maximum activity in the pH range between 3 and 5. Treatment of MBaL-30 and MBaL-60 with EDTA completely abolished their hemagglutinating activities. Addition of Zn and Fe ions to the ethylenediaminetetraacetic acid-treated MBaL-30 and MBaL-60 lectins did not only regained the loss of activity but also resulted in 200% to 300% increase in activity, respectively. MBaL-30 and -60 agglutinated gram positive Listeria monocytogenes and Staphylococcus aureus, whereas MBaL-30 could merely agglutinate Escherichia coli. None of these lectins could arrest bacterial growth. Addition of MBaL to cancer cell lines (Gastric cancer cell line (AGS) and Gastric cencer cell line (MKN45), Glioblastoma (ECV-304), and Human urinary bladder cancer cell line (U87-MG)) at varying concentrations did not cause statistically significant changes on cell growth and viability.


Assuntos
Momordica/metabolismo , Lectinas de Plantas/análise , Sementes/metabolismo , Sequência de Aminoácidos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Galactose/metabolismo , Testes de Hemaglutinação , Humanos , Peso Molecular , Lectinas de Plantas/química , Lectinas de Plantas/metabolismo
12.
Histochem Cell Biol ; 145(2): 185-99, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26553286

RESUMO

Recognition of glycans by lectins leads to cell adhesion and growth regulation. The specificity and selectivity of this process are determined by carbohydrate structure (sequence and shape) and topology of its presentation. The synthesis of (neo)glycoconjugates with bi- to oligo-valency (glycoclusters) affords tools to delineate structure-activity relationships by blocking lectin binding to an artificial matrix, often a glycoprotein, or cultured cell lines. The drawback of these assays is that glycan presentation is different from that in tissues. In order to approach the natural context, we here introduce lectin histochemistry on fixed tissue sections to determine the susceptibility of binding of two plant lectins, i.e., GSA-II and WGA, to a series of 10 glycoclusters. Besides valency, this panel covers changes in the anomeric position (α/ß) and the atom at the glycosidic linkage (O/S). Flanked by cell and solid-phase assays with human tumor lines and two mucins, respectively, staining (intensity and profile) was analyzed in sections of murine jejunum, stomach and epididymis as a function of glycocluster presence. The marked and differential sensitivity of signal generation to structural aspects of the glycoclusters proves the applicability of this method. This enables comparisons between data sets obtained by using (neo)glycoconjugates, cells and the tissue context as platforms. The special advantage of processing tissue sections is the monitoring of interference with lectin association at sites that are relevant for functionality. Testing glycoclusters in lectin histochemistry will especially be attractive in cases of multi-target recognition (glycans, proteins and lipids) by a tissue lectin.


Assuntos
Glicoconjugados/química , Glicoconjugados/farmacologia , Lectinas de Plantas/análise , Lectinas de Plantas/metabolismo , Animais , Sítios de Ligação/efeitos dos fármacos , Células CHO , Células CACO-2 , Cricetulus , Relação Dose-Resposta a Droga , Glicoconjugados/síntese química , Histocitoquímica , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ligação Proteica/efeitos dos fármacos , Relação Estrutura-Atividade
13.
Gastric Cancer ; 19(2): 531-542, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25840959

RESUMO

BACKGROUND: Although various molecular profiling technologies have the potential to predict specific tumor phenotypes, the comprehensive profiling of lectin-bound glycans in human cancer tissues has not yet been achieved. METHODS: We examined 242 advanced gastric cancer (AGC) patients without or with lymph node metastasis-N0 (n = 62) or N+ (n = 180)-by lectin microarray, and identified the specific lectins highly associated with AGC phenotypes. RESULTS: In seven gastric cancer cell lines, in contrast to expressed-in-cancer lectins, not-expressed-in-cancer (NEC) lectins were tentatively designated by lectin microarray. Binding signals of the specific lectins were robustly reduced in AGC patients with N+ status as compared with those with N0 status. The receiver operating characteristic curve determined the optimal cutoff value to differentiate N0 status from N+ status, and subsequent profiling of NEC lectins identified Vicia villosa agglutinin (VVA) association with the significant other lectins involved in lymph node metastasis. VVA reaction was clearly found on cancer cells, suggesting that it may result from carcinoma-stroma interaction in primary AGC, because VVA is an NEC lectin. Most intriguingly, VVA reaction was remarkably attenuated in the tumor cells of the metastatic lymph nodes, even if it was recognized in primary AGC. In AGC, histological type was strongly associated with soybean agglutinin and Bauhinia purpurea lectin, whereas p53 mutation was the best correlated with Griffonia simplicifolia lectin II. CONCLUSIONS: Lectin microarrays can be used to very accurately quantify the reaction of glycans with tumor tissues, and such profiles may represent the specific phenotypes, including N+ status, histological type, or p53 mutation of AGC.


Assuntos
Lectinas/metabolismo , Análise Serial de Proteínas/métodos , Neoplasias Gástricas/patologia , Idoso , Linhagem Celular Tumoral , Feminino , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patologia , Humanos , Lectinas/análise , Metástase Linfática/patologia , Masculino , Pessoa de Meia-Idade , Lectinas de Plantas/análise , Lectinas de Plantas/metabolismo , Proteínas de Soja/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Proteína Supressora de Tumor p53/genética
14.
Nutr Cancer ; 67(3): 401-10, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25710272

RESUMO

Legumes in combination with other products are the staple food for a large part of the world population, especially the low-income fragment, because their seeds provide valuable amounts of carbohydrates, fiber, and proteins, and have an important composition of essential amino acids, the sulphured amino acids being the limiting ones. Furthermore, legumes also have nonnutritional compounds that may decrease the absorption of nutrients or produce toxic effects; however, it has been reported that depending on the dose, these nonnutritional compounds also have different bioactivities as antioxidant, hypolipidemic, hypoglycemic, and anticarcinogenic agents, which have been proven in scientific studies. It has been observed that in countries with a high consumption of legumes, the incidence of colorectal cancer is lower. Some studies have shown that legume seeds are an alternative chemopreventive therapy against various cancers especially colon; this was verified in various animal models of induced by azoxymethane, a colon specific carcinogenic compound, in which a diet was supplemented with different concentrations of beans, lentils, chickpeas, or soybeans, mostly. These studies have proven the anticancer activity of legumes in early stages of carcinogenesis. Therefore, it is important to review the information available to elucidate the chemopreventive mechanisms of action of legume compounds.


Assuntos
Anticarcinógenos/farmacologia , Fabaceae/química , Isoflavonas/análise , Fenóis/análise , Ácido Fítico/análise , Fitoterapia , Lectinas de Plantas/análise , Proteínas de Plantas/análise , alfa-Amilases/análise
15.
Rinsho Byori ; 63(9): 1039-43, 2015 Sep.
Artigo em Japonês | MEDLINE | ID: mdl-26731891

RESUMO

Wisteria floribunda agglutinin (WFA) -positive human Mac-2-binding protein (WFA(+)-M2BP) is a new glycol marker related to liver fibrosis. The aim of the present study was to evaluate WFA(+)-M2BP as a predictor of liver fibrosis and hepatocellular carcinoma (HCC) development in patients with chronic hepatitis C. A total of 115 patients with chronic hepatitis C who underwent liver biopsy were included, and the diagnostic accuracy of WFA(+)-M2BP for liver fibrosis was evaluated. A case-control study was conducted including 14 patients with chronic hepatitis C who developed HCC and 52 controls to estimate the utility of WFA(+)-M2BP as a predictor of HCC development. WFA(+)-M2BP increased stepwisely with the progression of liver fibrosis (p < 0.001). The cumulative incidence of HCC development was significantly higher in patients with WFA(+)-M2BP ≥ 4.2 (p < 0.001) or in those with time-course changes in WFA(+)-M2BP (ΔWFA(+)-M2BP/year) =0.3(p=0.03). Multivariate analyses demonstrated that WFA(+)-M2BP ≥ 4.2 [hazard ratio (HR): 4.1, p = 0.04], AWFA'-M2BP/year ≥ 0.3 (HR: 5.5, p=0.008), and AFP ≥ 10 ng/mL (HR: 4.7, p = 0.03) were independent predictive factors of HCC development. Based on these data, we developed a simple scoring system to predict HCC development using these three factors. Using these scores, patients were classified into four groups; the cumulative incidence of HCC development significantly increased with increasing scores (p < 0.001). WFA(+)-M2BP measurements were useful as a predictor of liver fibrosis and HCC development. Furthermore, real-time monitoring of WFA(+)-M2BP may be a novel predictor of HCC development.


Assuntos
Antígenos de Neoplasias/análise , Carcinoma Hepatocelular , Hepatite C Crônica/patologia , Cirrose Hepática , Neoplasias Hepáticas , Glicoproteínas de Membrana/análise , Lectinas de Plantas/análise , Receptores de N-Acetilglucosamina/análise , Biópsia , Carcinoma Hepatocelular/etiologia , Feminino , Hepatite C Crônica/complicações , Humanos , Cirrose Hepática/etiologia , Neoplasias Hepáticas/etiologia , Masculino , Pessoa de Meia-Idade
16.
Braz. j. pharm. sci ; 51(3): 755-761, July-Sept. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-766318

RESUMO

Lectins have been described as glycoproteins that reversibly and specifically bind to carbohydrates. Legume lectins isolated from the subtribe Diocleinae (Canavalia, Dioclea andCratylia) are structurally homologous with respect to their primary structures. The Diocleinae lectins of Canavalia brasiliensis, Dioclea guianensis andCanavalia ensiformis have been shown to distinctly alter physiological parameters in isolated rat kidneys. Thus, the aim of this study was to investigate the effect of Cratylia floribunda lectin (CFL) on renal hemodynamics and ion transport in rats. In isolated perfused kidneys, CFL (10 mg/mL, n=5) increased RPP, RVR and decreased %TK+, but did not change urinary flow, glomerular filtration rate, sodium or chloride tubular transport. In isolated perfused mesenteric bed, CFL (3 and 10 mg/mL/min; n=4) did not alter tissue basal tonus or tissue contraction by phenylephrine (1 mM/mL/min). In conclusion, the seed lectin of Cratylia floribunda increased renal hemodynamic parameters showing a kaliuretic effect. This effect could be of tubular origin, rather than a result from haemodynamic alterations.


As lectinas são descritas como (glico)proteínas que se ligam, especificamente e reversivelmente, a carboidratos. Lectinas de leguminosas isoladas da subtribo Diocleinae (Canavalia, Dioclea eCratylia) são estruturalmente homólogas em relação às suas estruturas primárias. Demonstrou-se que as lectinas de DiocleinaeCanavalia brasiliensis, Dioclea guianensis eCanavalia ensiformis alteram diferentemente parâmetros fisiológicos em rins isolados de ratos. Dessa maneira, o objetivo deste estudo foi investigar o papel da lectina de Cratylia floribunda (CFL) na hemodinâmica renal e no transporte de íons em ratos. Em rins isolados perfundidos, CFL (10 mg/mL, n=5) aumentou a pressão de perfusão renal, a resistência vascular renal e reduziu o percentual do transporte tubular de K+, mas não alterou o fluxo urinário, a taxa de filtração glomerular e o percentual de transporte tubular dos íons sódio e cloreto. No leito mesentérico isolado perfundido, CFL (3 e 10 mg/mL/min, n=4) não alterou o tônus basal ou a contração do tecido induzida por fenilefrina (1 mM/mL/min). Em conclusão, a lectina de sementes de Cratylia floribunda altera parâmetros hemodinâmicos renais, provavelmente de origem tubular, e não por alterações hemodinâmicas.


Assuntos
Ratos , Transporte de Íons , Lectinas de Plantas/análise , Dioclea , Hemodinâmica , Amilorida/análise
17.
São Paulo; s.n; s.n; dez. 2014. 88 p. tab, graf, ilus.
Tese em Português | LILACS | ID: biblio-836756

RESUMO

Lectinas são proteínas cuja principal característica é a de se ligar específica e reversivelmente a carboidratos. BanLec é a lectina presente na polpa de bananas, que se liga especificamente a manose e glicose, e é capaz de induzir a proliferação de células T, podendo estimular a resposta imune. Existem indícios de que o teor de BanLec pode variar dependendo do estádio de amadurecimento e do tipo de cultivar, o que pode afetar a quantidade de BanLec existente na fruta quando consumida in natura e a possível resposta imune frente ao consumo de banana. Por este motivo, um dos objetivos desse trabalho foi determinar os teores e a atividade hemaglutinante de BanLec em extratos de farinha de banana verde, além de bananas das cultivares Pacovan, Figo, Terra, Mysore e Nanicão, nos estádios de maturação verde e maduro, e submetidas a tratamento com 1-MCP e baixa temperatura (para cv. Nanicão). Com vista a atender ao objetivo de avaliar seus efeitos imunomoduladores in vivo, a BanLec foi purificada da cultivar Nanicão e administrada por via oral a camundongos BALB/c. Os ensaios de atividade hemaglutinante dos extratos de banana apontaram para maior quantidade de BanLec no fruto maduro, quando comparado ao verde, e ausência dessa proteína na cultivar Figo. Os parâmetros imunológicos analisados após administração de BanLec aos camundongos demonstram que a resposta imune gerada após ingestão de BanLec é dose dependente, além disso, a administração de 50 µg de BanLec aos animais foi capaz de modular citocinas importantes na resposta imunológica, provavelmente causando um efeito que pode ser interpretado como mais protetor do que patogênico. Com base nos resultados obtidos, podemos concluir que existem diferenças nos teores de BanLec dependendo da cultivar e estádio de maturação analisado, sendo que essa proteína não está presente na polpa de todas as variedades de banana e finalmente, que ela tem grande potencial imunomodulador in vivo, uma vez que ativou citocinas de resposta anti-inflamatória


Lectins are proteins which bind specifically and reversibly to carbohydrates. BanLec is the lectin present in banana pulp, and it binds to mannose and glucose, being capable of inducing T-cell proliferation, and to stimulate the immune response. There are some evidence that the amount of BanLec may vary depending on the maturation stage of the fruit and the cultivar (cv.), which may affect the amount of BanLec and the possible immune response after consumption of banana. Thus, this study aimed to evaluate the amount of BanLec and its hemagglutinating activity in crude extracts of bananas from cultivars Pacovan, Figo, Terra, Mysore and Nanicão, in both unripe and ripe maturation stage, and also fruits which were treated with 1-MCP and low temperature. In addition, in order to access their immunomodulatory effects in vivo, BanLec was purified by affinity chromatography and administered orally to BALB/c mice. The hemagglutinating activity assays indicate higher amount of BanLec in ripe fruit. Moreover, the possible was undetectable in the pulp of banana Figo. The immunological parameters of mice orally fed with BanLec showed that the immunological response is dependent on the amount of protein administrated, in agreement to previous in vitro studies. Besides, 50 µg of BanLec, were able to modulate some cytokines in immune response, causing an effect that seems to be more protective than pathogenic. We conclude that there are important differences in amount of BanLec depending on the cultivar and the maturation stage, and BanLec has a dose-dependent immunomodulatory effect in vivo


Assuntos
Musa/imunologia , Lectinas de Plantas/análise , Imunomodulação/imunologia , Fatores Imunológicos/farmacologia , Bioquímica , Testes Imunológicos , Análise de Alimentos/métodos
18.
ScientificWorldJournal ; 2014: 978525, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25133268

RESUMO

Extracts from natural plants have been used in traditional medicine for many centuries worldwide. Artocarpus communis is one such plant that has been used to treat liver cirrhosis, hypertension, and diabetes. To our knowledge, this study is the first to investigate the antihepatoma activity of A. communis toward HepG2 and PLC/PRF/5 cells and the first to explore the relationship between antihepatoma activity and the active compound artocarpin content in different fractions of A. communis. A. communis methanol extract and fractions induced dose-dependent reduction of tumor cell viability. DNA laddering analysis revealed that A. communis extract and fractions did not induce apoptosis in HepG2 and PLC/PRF/5 cells. Instead, acridine orange staining revealed that A. communis triggered autophagic cell death in a dose-dependent manner. The antihepatoma activity of A. communis is attributable to artocarpin. The fractions with the highest artocarpin content were also the fractions with the highest antihepatoma activity in the following order: dichloromethane fraction > methanol extract > ethyl acetate fraction > n-butanol fraction > n-hexane fraction. Taken together, A. communis showed antihepatoma activity through autophagic cell death. The effect was related to artocarpin content. Artocarpin could be considered an indicator of the anticancer potential of A. communis extract.


Assuntos
Antineoplásicos/farmacologia , Artocarpus/química , Lectinas de Ligação a Manose/farmacologia , Extratos Vegetais/farmacologia , Lectinas de Plantas/farmacologia , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Células Hep G2 , Humanos , Lectinas de Ligação a Manose/análise , Extratos Vegetais/química , Lectinas de Plantas/análise
19.
Plant Foods Hum Nutr ; 69(2): 168-74, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24793353

RESUMO

Dwarf elder (Sambucus ebulus L.) berries are rich in health-promoting phytochemicals such as polyphenols and anthocyanins, and display a significant antioxidant activity. They are also rich in two lectins (ebulin f and SELfd) that share amino acid sequence homology with the elderberry allergen Sam n1 present in Sambucus nigra pollen and fruits. Ebulin f displays toxicity by oral ingestion. This study was aimed at eliminating the toxicity of these lectins whilst having little or no effect on the antioxidant properties of dwarf elder berries. We thus investigated the potential effects of incubation in a boiling water bath of extracts from several parts of the plant on total polyphenol content, antioxidant activity, total anthocyanins, cyanidin-3-glycoside content, and the sensitivity of purified dwarf elder fruit lectins to a simulated gastric fluid. The study shows that five minutes of said heat treatment fully sensitized both lectins to pepsin digestion, whilst minimally reducing phenol and antioxidant as well as free radical scavenging activities to below 13%. It proved possible to eliminate the potential risks derived from the presence of lectins in dwarf elder juices without any significant reduction in the content of the antioxidant compounds. Dwarf elder berries may thus be a valuable nutritional source.


Assuntos
Antocianinas/análise , Tecnologia de Alimentos/métodos , Lectinas de Plantas/análise , Polifenóis/análise , Sambucus/química , Antioxidantes/análise , Antioxidantes/farmacologia , Frutas/química , Frutas/crescimento & desenvolvimento , Glicosídeos/análise , Calefação , Pepsina A/química , Lectinas de Plantas/química , Lectinas de Plantas/toxicidade , Plantas Medicinais/química , Proteínas Inativadoras de Ribossomos Tipo 2/química
20.
Braz. j. biol ; 74(2): 489-492, 5/2014. tab, graf
Artigo em Inglês | LILACS | ID: lil-719244

RESUMO

Parkia pendula (Willd.) Walp. (Fabaceae) is a neotropical species of the genus Parkia more abundantly distributed in Central to South America. From the seeds of P. pendula a glucose/mannose specific lectin (PpeL) was isolated that has been characterised and used as a biotechnological tool but until now this is the first manuscript to analyse P. pendula mRNA expression in seedlings. For this porpoise a Differential display reverse transcription polimerase chain reaction (DDRT-PCR) was used to evaluate the expression of P. pendula lectin mRNAs in non-rooted seedlings. No bands were observed in the agarose gel, indicating the absence of mRNA of PpeL seedlings. our findings confirm that lectins mRNAs are differently regulated among species even if they are grouped in the same class.


Parkia pendula (Willd.) Walp. (Fabaceae) é a espécie neotropical do gênero Parkia mais abundantemente distribuída na América Central a do Sul. Das sementes de P. pendula foi isolada uma lectina glicose/manose específica (Ppel) que foi caracterizada e usada como ferramenta biotecnológica, porém até o momento esse é o primeiro artigo a analisar a expressão do mRNA nas plântulas de P. pendula. Para esse propósito uma reação de PCR diferencial de transcriptase reversa (DDRT-PCR) foi utilizada para avaliar a expressão do mRNA da lectina de P. pendula em plântulas não enraizadas. Nenhuma banda foi observada no gel de agarose, indicando a ausência de mRNA das plântulas de PpeL. Nossos achados confirmam que os mRNAs de lectinas são regulados de forma diferentes entre as espécies, mesmo que sejam agrupadas na mesma classe.


Assuntos
Fabaceae/genética , Lectinas de Plantas/genética , RNA Mensageiro/análise , RNA de Plantas/análise , Fabaceae/química , Lectinas de Plantas/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Plântula
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