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1.
Polymers (Basel) ; 14(16)2022 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-36015599

RESUMO

Bone tissue attracts cancer cell homing biologically, mechanically, or chemically. It is difficult and time consuming to identify their complex cross-talk using existed methods. In this study, a multi-component bone matrix was fabricated using gelatin, hydroxyapatite (HAp), and epidermal growth factor (EGF) as raw materials to investigate how "acellular" bone matrix affects cancer cell homing in bone. Then, EGF-responsive cancer cells were cultured with the scaffold in a dynamical bioreactor. For different culture periods, the effects of HAp, gelatin, and EGF on the cell adhesion, proliferation, 3D growth, and migration of cancer were evaluated. The results indicated that a small amount of calcium ion released from the scaffolds accelerated cancer MDA-MB-231 adhesion on the surface of inner pores. Moreover, degradable gelatin key caused cancer cell growth on the scaffold surface to turn into a 3D aggregation. Despite this, the formation of cancer spheroids was slow, and required 14 days of dynamic culture. Thankfully, EGF promoted cancer cell adhesion, proliferation, and migration, and cancer spheroids were observed only after 3-day culture. We concluded that the combination of the multiple components in this scaffold allows cancer cells to meet multiple requirements of cancer dynamic progression.

2.
Biosensors (Basel) ; 11(7)2021 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-34356691

RESUMO

The issue of micro-plastics is becoming more and more important due to their ubiquity and the harm they cause to the human body. Therefore, evaluating the biological-physical interaction of micro-plastics with health cells has become the focus of many research efforts. This study focuses on the movement mode and low concentration detection development for micro-plastics in surface plasmon resonance (SPR). Firstly, 20-micrometer micro-plastics were prepared by grinding and filtering, and the movement mode was explored; then, the characteristics were investigated by SPR. Chromatographic analysis showed that the surface charge of micro-plastics dominated the elution time, and estrogen receptors (ERs) played a supporting role. A difference of micro-plastics in SPR sensorgram was observed, inferring the micro-plastics' movement in rolling mode on the ERs. Characteristics analysis indicated that the low particle number of micro-plastics on SPR showed a linear relationship with the response unit (RU). When ERs were immobilized on the biosensor, the force of the binding of micro-plastics to ERs under an ultra-low background was equivalent to the dissociation rate constant shown as follows: PS (0.05 nM) > PVC (0.09 nM) > PE (0.14 nM). The ELISA-like magnetic beads experiment verified the specificity between ERs and micro-plastics. Therefore, by using the SPR technique, a biological-derived over-occupation of PS was found via higher binding force with ERs and longer retention time. In the future, there will be considerable potential for micro-plastics issues, such as identification in natural samples, biomarking, real-time detection in specific environments/regions and human health subject.


Assuntos
Monitoramento Ambiental , Microplásticos , Ressonância de Plasmônio de Superfície , Técnicas Biossensoriais , Desenho de Equipamento , Humanos
3.
Polymers (Basel) ; 13(15)2021 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-34372154

RESUMO

In the current study, we designed four cyclic peptide analogues by incorporating two cysteine residues in a BMP-2 linear knuckle epitope in such a way that the active region of the peptide could be either inside or outside the cyclic ring. Bone morphogenetic protein receptor BMPRII was immobilized on the chip surface, and the interaction of the linear and cyclic peptide analogues was studied using surface plasmon resonance (SPR). From the affinity data, the peptides with an active region inside the cyclic ring had a higher binding affinity in comparison to the other peptides. To confirm that our affinity data are in line in vitro, we studied the expression levels of RUNX2 (runt-related transcription factor) and conducted an osteogenic marker alkaline phosphatase (ALP) assay and staining. Based on the affinity data and the in vitro experiments, peptide P-05 could be a suitable candidate for osteogenesis, with higher binding affinity and increased RUNX2 and ALP expression in comparison to the linear peptides.

4.
Bioprocess Biosyst Eng ; 36(11): 1745-51, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23568753

RESUMO

High-purity fructooligosaccharides (FOS) were produced from sucrose by an innovative process incorporating immobilized Aspergillus japonicus and Pichia heimii cells. Intracellular FTase of A. japonicus converted sucrose into FOS and glucose, and P. heimii fermented glucose mainly into ethanol. The continuous production of FOS was carried out using a tanks-in-series bioreactor consisting of three stirred tanks. When a solution composed of 1 g L(-1) yeast extract and 300 g L(-1) sucrose was fed continuously to the bioreactor at a dilution rate of 0.1 h(-1), FOS at a purity of up to 98.2 % could be achieved and the value-added byproduct ethanol at 79.6 g L(-1) was also obtained. One gram of sucrose yielded 0.62 g FOS and 0.27 g ethanol. This immobilized dual-cell system was effective for continuous production of high-purity FOS and ethanol for as long as 10 days.


Assuntos
Aspergillus/metabolismo , Reatores Biológicos , Oligossacarídeos/biossíntese , Pichia/metabolismo , Cromatografia Líquida de Alta Pressão
5.
Enzyme Microb Technol ; 50(6-7): 287-92, 2012 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-22500894

RESUMO

Bostrycin, a red antibacterial agent with tetrahydroanthraquinone structure, has been isolated from Nigrospora sp. No. 407. This study investigated the potential antibacterial and multifunctional properties of matrixes through immobilization of bostrycin on their surface for immobilization of protein and prevention of bacterial growth. Bostrycin was immobilized on nonwoven polypropylene (PP) fabric by a technique using glutaraldehyde and polyethyleneimine for the activation of the surface. Glucose oxidase immobilized on bostrycin-treated nonwoven PP fabric showed high activity. The immobilization process improved thermal stability of the enzymes. During repeated assay for 30 cycles, the enzyme activity dropped to only 70% of the initial activity. Both bostrycin-treated nonwoven PP fabric sample and subsequently immobilized glucose oxidase sample on the surface also still exhibited a bacteriostatic effect. This is the first study to show that bostrycin is a promising coupling agent for surface modification on matrix and its potential applications in protein immobilization and biomaterial-centered infection.


Assuntos
Antraquinonas/química , Antibacterianos/farmacologia , Ascomicetos/enzimologia , Enzimas Imobilizadas/química , Glucose Oxidase/química , Têxteis , Antraquinonas/metabolismo , Antraquinonas/farmacologia , Antibacterianos/biossíntese , Antibacterianos/química , Biotecnologia/métodos , Clostridium botulinum/efeitos dos fármacos , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Glucose Oxidase/metabolismo , Testes de Sensibilidade Microbiana , Polipropilenos/química , Staphylococcus aureus/efeitos dos fármacos , Propriedades de Superfície
6.
J Biomed Biotechnol ; 2011: 359042, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21904443

RESUMO

In order to enhance the sensitivity of diagnosis, a recombinant clone containing domain I of HCV core (amino acid residues 1 to 123) was subjected to random mutagenesis. Five mutants with higher sensitivity were obtained by colony screening of 616 mutants using reverse ELISA. Sequence analysis of these mutants revealed alterations focusing on W(84), P(95), P(110), or V(129). The inclusion bodies of these recombinant proteins overexpressed in E. coli BL21(DE3) were subsequently dissolved using 6 M urea and then refolded by stepwise dialysis. Compared to the unfolded wild-type antigen, the refolded M3b antigen (W(84)S, P(110)S and V(129)L) exhibited an increase of 66% antigenicity with binding capacity of 0.96 and affinity of 113 µM(-1). Moreover, the 33% decrease of the production demand suggests that M3b is a potential substitute for anti-HCV antibody detection.


Assuntos
Anticorpos Anti-Hepatite C/análise , Antígenos da Hepatite C/genética , Antígenos da Hepatite C/imunologia , Hepatite C/diagnóstico , Proteínas do Core Viral/genética , Proteínas do Core Viral/imunologia , Sequência de Aminoácidos , Antígenos , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Humanos , Corpos de Inclusão/química , Dados de Sequência Molecular , Mutagênese , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia
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