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1.
J Environ Manage ; 360: 121108, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38754189

RESUMO

A novel air-lifting loop reactor combines anoxic, oxic, and settling zones to achieve organic and nutrient removal, as well as solid-liquid separation. To address sludge settling ability and operation stability issues caused by low dissolved oxygen in aerobic zones, this study proposes using modified polypropylene carriers to establish a fixed-film activated sludge (IFAS) system. A pilot-scale demonstration of the IFAS-based air-lifting loop reactor is conducted, and the results show successful operation for approximately 300 days. The pilot-scale reactor achieves a maximum aerobic granulation ratio of 16% in the bulk liquid. The IFAS system contributes to efficient removal of organic matter (96%) and nitrogen (94%) by facilitating simultaneous nitrification and denitrification, as well as fast solid-liquid separation with a low sludge volume index of 34 mL/g. Microbial analysis reveals enrichment of functional bacteria involved in nitrification, denitrification, and flocculation throughout the operation process.

2.
Foods ; 13(6)2024 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-38540870

RESUMO

Enzyme-assisted ultrasonic extraction (EAUE) was utilized and optimized for extracting polysaccharides from Schizochytrium limacinum meal (SLMPs) via the response surface methodology. The optimal EAUE conditions were determined as follows: enzyme concentration at 5.18%, ultrasonic temperature at 53 °C, ultrasonic duration of 40 min, ultrasonic power at 60 W, and a liquid-to-material ratio of 34 mL/g, achieving a polysaccharide extraction yield of 11.86 ± 0.61%. The purified polysaccharide component, SLMP1-1, isolated using DEAE Sepharose Fast Flow and Sephadex G-100 columns, exhibited potent antioxidant activity. SLMP1-1, with a molecular weight of 25.5 kDa, comprises glucose, mannose, arabinose, and galactose in a molar ratio of 16.39:14.75:1:693.03. 1H NMR analysis revealed the α configuration of SLMP1-1. Antioxidant assessments, including DPPH, ABTS, and ferric ion reduction assays, were detected with inhibitory values at 21.82-82.98%, 38.21-98.46%, and 3.30-20.30% at 0.2-1.0 mg/mL. This confirmed the effective antioxidant capacity of SLMP1-1, which is notably enhanced post oral and gastric digestion. The findings suggest that polysaccharides extracted from Schizochytrium limacinum meal hold significant promise as natural antioxidants.

3.
Biomed Pharmacother ; 172: 116223, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38325266

RESUMO

Trichinella spiralis is recognized for its ability to regulate host immune responses. The serine protease inhibitor of T. spiralis (Ts-SPI) participates in T. spiralis-mediated immunoregulatory effects. Studies have shown that helminth therapy exhibits therapeutic effects on metabolic diseases. In addition, we previously found that T. spiralis-derived crude antigens could alleviate diet-induced obesity. Thus, Ts-SPI was hypothesized to alleviate non-alcoholic fatty liver disease (NAFLD). Herein, recombinant Ts-SPI (rTs-SPI) was prepared from the muscle larvae T. spiralis. The relative molecular mass of rTs-SPI was approximately 35,000 Da, and western blot analysis indicated good immunoreactivity. rTs-SPI ameliorated hepatic steatosis, inflammation, and pyroptosis in NAFLD mice, which validated the hypothesis. rTs-SPI also reduced macrophage infiltration, significantly expanded Foxp3+ Treg population, and inactivated TLR4/NF-κB/NLRP3 signaling in the liver. Furthermore, rTs-SPI treatment significantly shifted the gut microbiome structure, with a remarkable increase in beneficial bacteria and reduction in harmful bacteria to improve gut barrier integrity. Finally, Abx-treated mice and FMT confirmed that gut-liver crosstalk contributed to NAFLD improvement after rTs-SPI treatment. Taken together, Taken together, these findings suggest that rTs-SPI exerts therapeutic effects in NAFLD via anti-inflammatory activity and gut-liver crosstalk.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Serpinas , Trichinella spiralis , Animais , Camundongos , Inibidores de Serina Proteinase , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Músculos , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico
4.
Foods ; 12(24)2023 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-38137229

RESUMO

Inhibiting α-amylase can lower postprandial blood glucose levels and delay glucose absorption, offering an effective approach for the development of antidiabetic diets. In this study, an active constituent with inhibitory activity against α-amylase was isolated and purified by bioassay-guided fractionation from Carya cathayensis Sarg. peel (CCSP). The active constituent was identified by NMR and Q-Exactive Orbitrap Mass Spectrometry as 5-O-p-coumaroylquinic acid (5-CQA). 5-CQA possessed strong inhibitory activity against α-amylase, with an IC50 value of 69.39 µM. In addition, the results of the kinetic study indicated that 5-CQA was a potent, reversible, noncompetitive inhibitor against α-amylase. The findings indicate that 5-CQA derived from CCSP has potential as a novel inhibitor against α-amylase, which can help mitigate postprandial blood sugar spikes, making it suitable for inclusion in antidiabetic diets.

5.
Bioresour Technol ; 384: 129306, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37328012

RESUMO

Achieving simultaneous carbon and nitrogen removal with sludge-liquid separation in a single reactor offers a solution to land shortages and improves treatment efficiency in municipal wastewater treatment plants of megacities. This study proposes a novel air-lifting continuous-flow reactor configuration with an alternative-aeration strategy that creates multi-functional zones for anoxic, oxic, and settlement processes. The optimal operating conditions for the reactor include a long anoxic hydraulic retention time, low dissolved oxygen (DO) in the oxic zone, and no specific reflux for external nitrifying liquid, which exhibit a high nitrogen removal efficiency of over 90% in treating real sewage with C/N < 4 in the pilot-scale study. Results show that a high sludge concentration and a low DO concentration facilitate simultaneous nitrification and denitrification, and a well mixing of sludge and substrate in different reaction zones promotes mass transfer and microbial activity. The long-term operation enriches functional microbes for carbon storage and nutrient removal.


Assuntos
Esgotos , Águas Residuárias , Nitrificação , Desnitrificação , Oxigênio , Remoção , Reatores Biológicos , Nitrogênio/análise , Carbono , Eliminação de Resíduos Líquidos/métodos
6.
Food Microbiol ; 113: 104279, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37098435

RESUMO

Integrons play a pivotal role in the dissemination of antimicrobial resistance, because they can capture and express exogenous antimicrobial resistance genes. This study aimed to elucidate the structure and contribution of different elements of class 2 integrons to fitness costs in their host bacteria and evaluate their adaptability to the "farm-to-table" process. We mapped 27 typical class 2 integrons of Escherichia coli isolated from aquatic foods and pork products, each harboring an inactive truncated class 2 integrase gene and the gene cassette (GC) array dfrA1-sat2-aadA1 with strong Pc2A/Pc2B promoters. Notably, the fitness costs associated with class 2 integrons depended on the Pc promoter strength and quantity and content of GCs in the array. Additionally, the costs of integrases were activity-dependent, and a balance was identified between GC capture ability and integron stability, which could explain the inactive truncated integrase identified. Although typical class 2 integrons exhibited low-cost structures in E. coli, the bacteria incurred biological costs, including decreasing growth rates and biofilm formation, in farm-to-table environments, especially under low-nutrient conditions. Nevertheless, sub-inhibitory antibiotic concentrations led to the selection of class 2 integron-carrying bacteria. This study provides important insights into how integrons may travel from preharvest to consumer goods.


Assuntos
Escherichia coli , Integrons , Integrons/genética , Fazendas , Antibacterianos/farmacologia , Bactérias , Integrases/genética , Farmacorresistência Bacteriana/genética
7.
Int Immunopharmacol ; 117: 109924, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36848791

RESUMO

Obesity, an increasingly prevalent disease worldwide, is accompanied by chronic inflammation and intestinal dysbiosis. Helminth infections have been increasingly proved to exhibit a protective role in several inflammation-associated diseases. Considering the side effects of live parasite therapy, efforts have been made to develop helminth-derived antigens as promising candidates with fewer adverse effects. This study aimed to evaluate the effect and mechanisms of TsAg (T. spiralis-derived antigens) on obesity and the associated inflammation in high-fat diet (HFD)-fed mice. C57BL/6J mice were fed a normal diet or HFD with or without TsAg treatment. The results reported that TsAg treatment alleviated body weight gain and chronic inflammation induced by HFD. In the adipose tissue, TsAg treatment prevented macrophage infiltration, reduced the expression of Th1-type (IFN-γ) and Th17-type (IL-17A) cytokines while upregulating the production of Th2-type (IL-4) cytokines. Furthermore, TsAg treatment enhanced brown adipose tissue activation and energy and lipid metabolism and reduced intestinal dysbiosis, intestinal barrier permeability and LPS/TLR4 axis inflammation. Finally, the protective role of TsAg against obesity was transmissible via the fecal microbiota transplantation approach. For the first time, our findings showed that TsAg alleviated HFD-induced obesity and inflammation via modulation of the gut microbiota and balancing the immune disorders, suggesting that TsAg might be a safer promising therapeutic strategy for obesity.


Assuntos
Dieta Hiperlipídica , Trichinella spiralis , Animais , Camundongos , Dieta Hiperlipídica/efeitos adversos , Disbiose/metabolismo , Camundongos Endogâmicos C57BL , Obesidade/metabolismo , Inflamação/tratamento farmacológico , Inflamação/complicações , Citocinas/uso terapêutico
8.
Cytokine ; 162: 156086, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36427469

RESUMO

BACKGROUND: Hypoxia is an important microenvironmental factor that induces Endometriosis (EMs), but its mechanism remains unclear. Our study aims to investigate the mechanisms of miR-150-5p on hypoxia-induced EMs. METHODS: Ovarian endometriosis cyst wall stromal cell lines CRL-7566 cells were treated with hypoxia. Cell migration ability was measured by Transwell assay. qRT-PCR was performed to detect miR-150-5p and PDCD4 expression. The autophagy-related proteins (LC3-I, LC3-II, Beclin-1, and p62), epithelial-mesenchymal transition (EMT) related proteins (E-cadherin, N-cadherin, and Vimentin) and NF-κB signaling pathway related proteins p65 expression were measured by western blot. Dual-luciferase reporter gene assay verified the binding relationship between miR-150-5p and PDCD4. RESULTS: After hypoxia treatment, the miR-150-5p expression was up-regulated in CRL-7566 cells, while the expression of PDCD4 was down-regulated. In CRL-7566 cells, autophagy, migration and EMT were increased after hypoxia treatment. The autophagy inhibitor 3-MA inhibited hypoxia-induced the autophagy, migration and EMT of CRL-7566 cells. Hypoxia-induced autophagy and EMT of CRL-7566 cells were inhibited after knocking down miR-150-5p. Then miR-150-5p negatively regulated PDCD4 expression. PDCD4 knockdown reversed the inhibitory effect of miR-150-5p silencing on hypoxia-induced autophagy and EMT of CRL-7566 cells. Inhibiting the NF-κB signaling pathway weakened the effect of PDCD4 knockdown on hypoxia-induced autophagy and EMT of CRL-7566 cells. CONCLUSION: MiR-150-5p silencing inhibited hypoxia-induced autophagy and EMT of endometriotic cells by regulating the PDCD4/NF-κB signaling pathway.


Assuntos
Endometriose , MicroRNAs , Feminino , Humanos , NF-kappa B/genética , NF-kappa B/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Transição Epitelial-Mesenquimal/genética , Endometriose/genética , Transdução de Sinais/genética , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Movimento Celular/genética , Autofagia/genética , Proliferação de Células , Hipóxia , Linhagem Celular Tumoral , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo
9.
Biol Trace Elem Res ; 201(9): 4508-4517, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36567423

RESUMO

The objective of this study was to investigate the effects of iron nanoliposomes on iron supplementation and toxicity in SD rats induced by a low-iron diet. The size and infrared spectroscopy of a liposomal oral delivery system were investigated. The particle size of nanoliposomes embedded with chelates was increased. Infrared spectra proved that peptides-iron and blank nanoliposomes were bonded by interaction forces, including the fracture of hydrogen bonds, C = C bonds, hydrophobic interaction, and C-N bonds. We found that iron supplementation chelates had a certain protective effect on viscera after being embedded by nanoliposomes. After 10 days of treatment, the concentration of hemoglobin could be gradually increased. Nanoliposome encapsulated peptides-iron has a better effect than other groups. At the same time, SOD, MDA, and CAT reached normal levels after 20 days. Histological results showed that the sections of the nanoliposomes groups were clearer than those of the other groups. There was a little inflammation in the liver without obvious pathological changes, which also proved that the iron chelates embedded by nanoliposomes had no obvious side effects on iron supplementation in rats. Nanoliposome encapsulated peptides-iron has a small side effect and a significant curative effect of iron supplementation. It maybe has a good application prospect in the clinical medical field.


Assuntos
Ferro , Lipossomos , Ratos , Animais , Ratos Sprague-Dawley , Lipossomos/química , Peptídeos/química , Quelantes de Ferro , Suplementos Nutricionais
10.
Artigo em Inglês | MEDLINE | ID: mdl-36193142

RESUMO

Objective: Intrauterine adhesion (IUA) represents an endometrial repair disorder that is associated with menstrual disorders, recurrent pregnancy loss, and infertility. This study aimed to explore the underlying biological mechanisms of Guishen Huoxue decoction for the treatment of IUA based on network pharmacology. Methods: The selection of active compounds for Guishen Huoxue decoction and prediction of relevant targets were performed by the TCMSP and Swiss Target Prediction databases, respectively. The targets of IUA were obtained by three databases, including Online Mendelian Inheritance in Man (OMIM), DisGeNET, and GeneCards. The drug-disease regulatory network was constructed via Cytoscape software, following the acquisition of common genes of active compounds of drug Guishen Huoxue decoction and disease IUA, which was carried out through Venny software. Protein-protein interaction (PPI) network and function enrichment analyses were performed. Results: According to the data obtained from TCMSP, a total of 200 potential active compounds of Guishen Huoxue decoction and their related targets (1068) were screened by the Swiss Target Prediction database. 1303 disease targets and 134 common targets were identified. The drug-disease regulatory network showed that 165 active compounds were found to be involved in the treatment of IUA. Among 134 common targets, AKT1, SRC, TP53, VEGFA, and IL-6 were predicted as core genes against IUA. PI3K-Akt, Rap1, Ras, and AGE-RAGE were the main signaling pathways that participated in the treatment of Guishen Huoxue decoction for IUA. Conclusion: The active compounds of Guishen Huoxue decoction confer therapeutic effects against IUA by regulating fibrosis, inflammation, and oxidative stress through major signaling pathways such as PI3K-Akt and AGE-RAGE.

11.
J Immunol Res ; 2022: 4333128, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36249421

RESUMO

Objective: Thin endometrium is considered as a leading cause of infertility, recurrent pregnancy loss, and repeated implantation failure. The seed of Cuscutae Semen (CS) has been used to prevent aging and improve sexual function in Traditional Chinese Medicine. However, the pharmacological mechanism of CS in preventing and treating thin endometrium remains to be elucidated. Methods: Three public databases, TCMSP, GeneCards, and OMIM, were searched to collect the main active compounds and putative molecules of CS, as well as the targets of thin endometrium, respectively. The CS and thin endometrium common targets were subject to protein-protein interaction (PPI) analysis followed by functional enrichment analysis. The best binding mode of CS compounds and common target proteins was evaluated by molecular docking and analysis in the AutoDockTools. Results: In total, 11 main active compounds, 102 drug target proteins, and 70 CS and thin endometrium common targets were identified. There were 68 nodes with 722 edges in the PPI network; HIF1A, MYC, ESR1, and EGFR were the top 4 targets. After functional enrichment analysis, it was revealed that the therapeutic effects of active compounds of CS on thin endometrium were achieved through cellular response to chemical stress, transcription regulator, DNA-binding transcription factor binding, chemical carcinogenesis-receptor activation, lipid, and atherosclerosis. The molecular docking analysis revealed that the 3 active compounds of CS, quercetin, matrine, and isorhamnetin, have good binding ability with their targets, HIF1A, MYC, ESR1, and EGFR. Conclusion: Our study uncovers the main active compounds in CS and their corresponding targets related to thin endometrium which explains the pharmacological mechanism underlying therapeutic effects of CS on thin endometrium.


Assuntos
Medicamentos de Ervas Chinesas , Quercetina , DNA , Medicamentos de Ervas Chinesas/farmacologia , Endométrio , Receptores ErbB , Feminino , Humanos , Imunoterapia , Lipídeos , Medicina Tradicional Chinesa , Simulação de Acoplamento Molecular , Farmacologia em Rede , Gravidez , Fatores de Transcrição
12.
Front Microbiol ; 13: 951782, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36051758

RESUMO

Background: Polycystic ovary syndrome (PCOS) is a complex disease that afflicts women of reproductive age, and its pathological mechanism has not been well explained. The gut microbiota is believed to be closely related to the development of PCOS. Although an important component of the gut microbiome, the role of the gut virome in the development of PCOS is still unclear. Methods: In this study, we profiled and compared the gut viral community of 50 patients with PCOS and 43 healthy women based on the analysis of their fecal whole-metagenome dataset. Results: The gut virome of PCOS patients exhibited a significant decrease in within-sample viral diversity and a remarkable alteration of the overall virome composition compared with that of healthy controls. At the family level, Siphoviridae was significantly depleted in the gut virome of patients, while Quimbyviridae was enriched. We identified 1,089 viral operational taxonomic units (vOTUs) that differed in relative abundance between the two groups, of which 455 vOTUs were enriched in PCOS patients (including numerous Bacteroidaceae phages) and 634 were enriched in controls (including numerous viruses predicted to infect Oscillospiraceae, Prevotellaceae, and Ruminococcaceae). Functional comparison of the PCOS-enriched and control-enriched vOTUs uncovered the viral functional signatures associated with PCOS. Furthermore, we demonstrated gut viral signatures for disease discrimination and achieved an area under the receiver operator characteristic curve (AUC) of 0.938, demonstrating the potential of the gut virome in the prediction of PCOS. Conclusion: Our findings reveal specific alterations in viral diversity and taxonomic and functional compositions of the gut virome of PCOS patients. Further studies on the etiology of PCOS and the gut viral community will offer new prospects for treating and preventing PCOS and its related diseases.

13.
Int Immunol ; 34(7): 365-377, 2022 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-35485964

RESUMO

AIM: this study aimed to investigate the role of long non-coding RNA (lncRNA) epidermal growth factor receptor antisense RNA 1 (EGFR-AS1), an antisense transcript of EGFR, in leiomyosarcoma (LMS) and the underlying mechanisms. METHODS: levels of EGFR-AS1 and programmed death ligand 1 (PD-L1) were measured in LMS tissues and cell lines using quantitative real-time PCR (qRT-PCR), as well as western blotting and/or immunohistochemical staining; flow cytometry was employed to validate the role of EGFR-AS1 in altering the activity of CD8+ T cells; interaction of EGFR-AS1 and EGFR was determined by fluorescent in situ hybridization (FISH) and RNA pull-down; regulation of MYC on the PD-L1 promoter was assessed by chromatin immunoprecipitation (ChIP); a xenograft in vivo tumor growth assay was applied to verify the EGFR-AS1/EGFR/MYC/PD-L1 axis in vivo. RESULTS: up-regulation of EGFR-AS1 and PD-L1 in LMS tissues was negatively correlated with CD8+ T-cell infiltration; EGFR-AS1 positively regulated PD-L1, thereby strengthening interaction of LMS cells and CD8+ T cells and triggering CD8+ T cell apoptosis via the PD-1/PD-L1 checkpoint; EGFR-AS1 co-localized and interacted with EGFR to promote MYC activity; MYC was identified as a transcriptional activator of PD-L1. CONCLUSION: lncRNA EGFR-AS1 was demonstrated to increase PD-L1 expression through the EGFR/MYC pathway in LMS cells, thereby repressing T-cell infiltration and contributing to immune escape.


Assuntos
Leiomiossarcoma , RNA Longo não Codificante , Evasão Tumoral , Antígeno B7-H1 , Linhagem Celular Tumoral , Receptores ErbB/genética , Receptores ErbB/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Hibridização in Situ Fluorescente , Leiomiossarcoma/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo
14.
Food Chem ; 372: 131337, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-34818745

RESUMO

The process-related physicochemical, digestive and rheological properties of protein prepared by subcritical dimethyl ether extraction (SDEE) were comprehensively investigated and compared with those obtained by pH-shift, to study the industrial potential of SDEE. Two different materials from tuna (meat and liver) were studied in parallel, and SDEE had similar effects on the proteins in them. The protein component was almost unchanged before and after SDEE, while the content of water-soluble protein and alkali-soluble protein was substantially reduced and increased after pH-shift, respectively. We also found that SDEE had superior ability to pH-shift to conserve light metals, remove lipids and heavy metals, and maintain protein structure. Furthermore, SDEE-produced protein powders were easier for humans to digest, and their gelation and emulsification were also superior to those prepared by pH-shift. The aforementioned results suggest that SDEE can remove more impurities, and the obtained protein has outstanding potential in industrial applications.


Assuntos
Éteres Metílicos , Atum , Animais , Humanos , Extratos Vegetais , Reologia
15.
Front Microbiol ; 13: 1062577, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36687650

RESUMO

The worrying emergence of multiple resistance genes to last-resort antibiotics in food animals and human populations throughout the food chain and relevant environments has been increasingly reported worldwide. Enterobacteriaceae pathogens are considered the most common reservoirs of such antibiotic resistance genes (ARGs). Thus, a rapid, efficient and accurate detection method to simultaneously screen and monitor such ARGs in Enterobacteriaceae pathogens has become an urgent need. Our study developed a recombinase polymerase amplification (RPA) assay combined with a lateral flow dipstick (LFD) for simultaneously detecting predominant resistance genes to last-resort antibiotics of Enterobacteriaceae pathogens, including mcr-1, blaNDM-1 and tet(X4). It is allowed to complete the entire process, including crude DNA extraction, amplification as well as reading, within 40 min at 37°C, and the detection limit is 101 copies/µl for mcr-1, blaNDM-1 and tet(X4). Sensitivity analysis showed obvious association of color signals with the template concentrations of mcr-1, blaNDM-1 and tet(X4) genes in Enterobacteriaceae pathogens using a test strip reader (R 2 = 0.9881, R 2 = 0.9745, and R 2 = 0.9807, respectively), allowing for quantitative detection using multiplex RPA-LFD assays. Therefore, the RPA-LFD assay can suitably help to detect multiple resistance genes to last-resort antibiotics in foodborne pathogens and has potential applications in the field.

16.
Int J Biol Macromol ; 150: 1046-1052, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31743711

RESUMO

Excessive coagulation can easily lead to arterial and venous thrombosis, which is the main reason for the evolution of myocardial infarction and cerebrovascular accidents. As a key coagulation factor for the coagulation pathway, thrombin has become a remarkable target for the control of thrombosis. The synthesized peptide P13 with amino acid sequence of N-RGDAGFAGDDAPR was expected to be an inhibitor with higher antithrombotic activity. The results showed that the IC50 (50% inhibition of thrombin activity) of the peptide P13 was determined by colorimetric method to be 115 µM. And enzyme kinetic experiments showed that P13 was a competitive inhibitor of thrombin with Ki = 106 µM. Fluorescence spectra and three-dimensional fluorescence showed that P13 could alter the secondary structure of thrombin and the microenvironment of certain chromogenic amino acids. P13 can spontaneously bind with thrombin exosite 1 in the form of 1:1 mainly through hydrogen bonding and van der Waals force. And the optimal docking mode of P13 and thrombin was revealed by molecular docking with "-CDOCKER_Energy" of 178.679 kcal mol-1. This study revealed P13 may become a potential anticoagulant drug widely used after further studies in preclinical and clinical trials.


Assuntos
Antitrombinas/química , Peptídeos/química , Trombina , Antitrombinas/síntese química , Humanos , Peptídeos/síntese química , Trombina/antagonistas & inibidores , Trombina/química
17.
Int J Biol Macromol ; 140: 1116-1125, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31425762

RESUMO

This study aimed to obtain the purified fractions of Mytilus coruscus polysaccharides (MCPs) and investigate their antioxidant activities. MCPs were prepared through ultrasonic-assisted enzymatic extraction optimized by employing the response surface methodology. A single-factor experiment was conducted using the Box-Behnken design to determine the optimum extraction conditions of MCPs. The ultrasonic power was 60 W, liquid-to-material ratio was 30 mL/g, extraction time was 36 min, extraction temperature was 64 °C, enzyme concentration was 3.2%, and polysaccharide extraction yield was 12.86% ±â€¯0.12%. A novel polysaccharide (MCP1-2) was obtained after the purification with AB-8 macroporous resin, DEAE Sepharose Fast Flow, and Sepharose CL-6B column. The molecular weight of MCP1-2 was estimated to be 134.9 kDa according to high-performance gel permeation chromatography. High-pressure liquid-phase chromatography results showed that MCP1-2 contained mannose, rhamnose, glucuronic acid, glucose, galactose, and L-Fuc at a molar ratio of 1.53:1:4.83:81.82:2.36:1.51. Infrared and NMR spectroscopies confirmed that MCP1-2 possessed α- and ß- configurations. The antioxidant activities of MCP1-2 were investigated in vitro, and the results showed that MCP1-2 had good antioxidant activity and can be used as a natural antioxidant in food.


Assuntos
Antioxidantes/farmacologia , Enzimas/metabolismo , Mytilus/química , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Ultrassom , Animais , Espectroscopia de Ressonância Magnética , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Fatores de Tempo
18.
J Food Biochem ; 43(7): e12876, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31353718

RESUMO

This work was aimed at producing peptides containing iron-binding capabilities from scad (Decapterus maruadsi) processing by-product with alcalase hydrolysis. The chelating peptides were purified by ultrafiltration, immobilized-metal affinity chromatography, gel filtration chromatography, and reversed-phase high-performance liquid chromatography. A novel iron-binding peptide was purified with 1,386.63 Da molecular weight and amino acid sequence of QKGTYDDYVEGL. The peptide binds to iron mainly through carboxyl and hydroxyl oxygen bonds. The iron-binding peptide can significantly promote the absorption of inorganic iron in Caco-2 cells. These results have contributed to development of the peptide from scad processing by-products hydrolyzate in iron supplementations. PRACTICAL APPLICATIONS: Iron deficiency is one of the most common and widespread nutritional disorders in the world. Iron-peptide chelates may be suitable for iron-fortification. Our study shows that a peptide purified from scad processing by-product has iron-chelating activity, and significantly increases iron absorption by Caco-2 cells. Hence, this peptide has potential application as a novel carrier for enhancing iron absorption.


Assuntos
Quelantes de Ferro/química , Peptídeos/química , Resíduos/análise , Animais , Biocatálise , Células CACO-2 , Cromatografia de Afinidade , Cromatografia de Fase Reversa , Proteínas de Peixes/química , Humanos , Ferro/química , Quelantes de Ferro/isolamento & purificação , Peptídeos/isolamento & purificação , Subtilisinas/química
19.
Ecotoxicol Environ Saf ; 171: 683-690, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-30658304

RESUMO

Ozone layer depletion leads to elevated ultraviolet-B (UV-B) radiation, which affects plant growth; however, little is known about the relationship between root growth and signaling molecules in roots. Therefore, in this work, simulated UV-B radiation was used to study the effects of elevated UV-B radiation on root growth of soybean seedlings and changes in the content of signaling molecules in roots. The results showed that compared with the control, the 2.63 kJ m-2 d-1 and 6.17 kJ m-2 d-1 elevated UV-B radiation treatments inhibited root growth, and root growth parameters (total root length, root surface area, root volume, average diameter, root tip number, and root dry weight) all decreased. For root signaling molecules, the content of nitric oxide, reactive oxygen species, abscisic acid, salicylic acid, and jasmonic acid increased, and the content of auxin, cytokinin, and gibberellin decreased. The above indices changed more significantly under the 6.17 kJ m-2 d-1 treatment. After withdrawal of the exposure, the above indices could be restored to a certain extent. These data indicated that UV-B radiation interfered with root growth by affecting the content of signaling molecules in roots, and the degree of the effects was related to the intensity of UV-B radiation. The results from this study provide a theoretical basis for studying the preliminary mechanism of elevated UV-B radiation on root growth and possible pathways that can mitigate UV-B radiation damage for root growth. ONE SENTENCE SUMMARY: The effects of elevated UV-B on root growth of soybean seedlings were regulated by signaling molecules, and the degree of the effects was related to the intensity of UV-B radiation.


Assuntos
Glycine max/efeitos da radiação , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/efeitos da radiação , Transdução de Sinais/efeitos da radiação , Raios Ultravioleta , Ácido Abscísico/metabolismo , Ciclopentanos/metabolismo , Citocininas/metabolismo , Relação Dose-Resposta à Radiação , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Óxido Nítrico/metabolismo , Oxilipinas/metabolismo , Desenvolvimento Vegetal/efeitos da radiação , Reguladores de Crescimento de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ácido Salicílico/metabolismo , Plântula/efeitos dos fármacos , Plântula/metabolismo , Glycine max/metabolismo
20.
RSC Adv ; 9(32): 18498-18505, 2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35515240

RESUMO

Thrombin is a critical serine protease in the coagulation system and is widely used as a target protein for antithrombotics. Spectroscopic analysis is a simple and effective method that is used to study the interaction between small molecules and proteins. In this study, the interactions of a potential antithrombotic peptide AGFAGDDAPR (P10) with thrombin were investigated by fluorescence spectroscopy, UV-vis spectroscopy, circular dichroism, Fourier-transform infrared spectroscopy and Raman spectroscopy, respectively. The results showed that the peptide P10 bonded to thrombin via hydrogen bonding and van der Waals forces, resulting in fluorescence quenching. And, the secondary structure of thrombin changed, the ß-sheet decreased, and the random coil increased. The peptide P10 bonded to proline and lysine, and changed the space structure of thrombin, resulting in inhibition of thrombin activity. The results contributed to exploration of the mechanism of this potential antithrombotic drug interaction with thrombin in order to provide a preliminary understanding of the pharmacodynamic properties of P10.

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