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1.
J Sci Food Agric ; 97(10): 3133-3140, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27882566

RESUMO

BACKGROUND: Mixed larvae and pupae of weaver ant (Oecophylla smaragdina) are widely used as an important food ingredient in regions of Thailand. They have high nutritional values and comprise 53% protein and 13% lipid. Peptides derived from food proteins have been shown to possess biological activities. RESULTS: Peptides derived from pepsin and trypsin digestion of these weaver ant larvae and pupae were purified based on angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities, and their amino acid sequences were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In silico docking of peptides with ACE successfully predicted the inhibitory peptides as confirmed by their chemical synthesis. Two peptides with sequences of FFGT and LSRVP showed IC50 values for ACE inhibition of 19.5 ± 1.7 and 52.7 ± 4.0 µmol L-1 , respectively. In addition, one potent antioxidant peptide with a sequence of CTKKHKPNC showed IC50 values of 48.2 ± 2.1 µmol L-1 for DPPH assay and 38.4 ± 0.2 µmol L-1 for ABTS assay, respectively. CONCLUSION: These results indicate that proteins from larvae and pupae of weaver ants are potential sources of peptides with anti-ACE and antioxidation bioactivities. © 2016 Society of Chemical Industry.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/química , Antioxidantes/química , Formigas/química , Proteínas de Insetos/química , Sequência de Aminoácidos , Animais , Biocatálise , Humanos , Cinética , Larva/química , Pepsina A/química , Mapeamento de Peptídeos , Peptidil Dipeptidase A/química , Pupa/química , Espectrometria de Massas em Tandem , Tailândia
2.
J Virol Methods ; 188(1-2): 51-6, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23219929

RESUMO

Yellow head virus (YHV) is a highly virulent pathogen that has caused severe mortality in cultivated shrimp (Penaeus monodon and Penaeus vannamei) in Thailand. There are several technologies that are applied to detect YHV for further control of the disease. RT-PCR is currently widely used in the laboratory, but it has some disadvantages related to cost, time-consuming and complexity. An alternative assay combines RT with loop-mediated isothermal amplification (LAMP) that not only provides high specificity, sensitivity and rapidity, but is also cheaper and more suitable for field applications in shrimp aquaculture than the RT-PCR. RT-LAMP is performed under isothermal conditions with a set of four to six primers designed to recognize six to eight distinct target sequences, and it has been combined with a chromatographic lateral-flow dipstick (LFD) to detect LAMP amplified product, which avoids the use of gel electrophoresis. In this study, RT-LAMP for the detection of YHV was developed by isothermal amplification at 65 °C for 45 min, followed by hybridization with an FITC-labeled DNA probe for 5 min and detected by LFD within 5 min (time required approximately 55 min, excluding RNA extraction and preparation time). The detection limit of RT-LAMP-LFD was 0.1 pg RNA extracted from shrimp infected with YHV equivalent to the nested RT-PCR, and no cross reaction was observed with other common shrimp viral pathogens. The LAMP method described in this study showed a rapid, high sensitivity and specificity and it is recommended as user-friendly for diagnosis of YHV in the field.


Assuntos
Cromatografia de Afinidade/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Penaeidae/virologia , Roniviridae/isolamento & purificação , Medicina Veterinária/métodos , Animais , Aquicultura , Corantes Fluorescentes , Sondas de Oligonucleotídeos/genética , Sensibilidade e Especificidade , Temperatura , Tailândia
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