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1.
Metabolites ; 12(8)2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35893262

RESUMO

In 2020, approximately 9.3 billion tons of crustaceans were consumed, and 45-48% of shrimp shell (SS) by-products were discarded as waste. In this study, the SS of Litopenaeus vannamei was fermented by Lactobacillus plantarum LV33204, Stenotrophomonas maltophilia LV2122 (strong proteolytic activity), and Aeromonas dhakensis LV1111 (chitin-degrading activity), and the optimal fermentation conditions of liquid-fermented SS was established. Contents of total peptide, astaxanthin, and total phenolic content of the fermented SS were significantly higher than that of unfermented SS. In the presence of fermented SS, glucose uptake and insulin resistance of TNF-α-stimulated FL83B hepatocytes were markedly improved. Furthermore, daily oral supplement of fermented SS to streptozotocin (STZ)/nicotinamide (NA)-induced diabetic rats for 7 weeks significantly reduced plasma glucose and insulin resistance. Meanwhile, ingestion of fermented SS might enhance hepatic catabolism of glucose by increasing hexokinase and glucose-6-phosphate dehydrogenase activity and decreasing glucose-6-phosphatase activity. In addition, the fermented SS downregulated plasma total cholesterol (TG), triglycerides (TCs), low-density lipoprotein cholesterol (LDL-C), liver TG, and TC and lipid peroxidation levels in diabetic rats. In conclusion, a biorefinery process for waste SS was established through mixed strain fermentation. The in vitro and in vivo data reveal that the fermented SS is a promising functional food for the management of diabetic hyperglycemia and hyperlipidemia.

2.
J Vis Exp ; (110)2016 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-27167162

RESUMO

Surveillance using biomarkers is critical for the early detection, rapid intervention, and reduction in the incidence of diseases. In this study, we describe the preparation of polycrystalline silicon nanowire field-effect transistors (pSNWFETs) that serve as biosensing devices for biomarker detection. A protocol for chemical and biomolecular sensing by using pSNWFETs is presented. The pSNWFET device was demonstrated to be a promising transducer for real-time, label-free, and ultra-high-sensitivity biosensing applications. The source/drain channel conductivity of a pSNWFET is sensitive to changes in the environment around its silicon nanowire (SNW) surface. Thus, by immobilizing probes on the SNW surface, the pSNWFET can be used to detect various biotargets ranging from small molecules (dopamine) to macromolecules (DNA and proteins). Immobilizing a bioprobe on the SNW surface, which is a multistep procedure, is vital for determining the specificity of the biosensor. It is essential that every step of the immobilization procedure is correctly performed. We verified surface modifications by directly observing the shift in the electric properties of the pSNWFET following each modification step. Additionally, X-ray photoelectron spectroscopy was used to examine the surface composition following each modification. Finally, we demonstrated DNA sensing on the pSNWFET. This protocol provides step-by-step procedures for verifying bioprobe immobilization and subsequent DNA biosensing application.


Assuntos
Técnicas Biossensoriais/instrumentação , Nanofios/química , Silício/química , Transistores Eletrônicos , Técnicas Biossensoriais/métodos , DNA/análise , DNA/química , Sondas de DNA/química , Humanos , Espectroscopia Fotoeletrônica , Proteínas/análise , Proteínas/química
3.
Biochem Pharmacol ; 84(2): 224-31, 2012 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-22521946

RESUMO

Cellular redox signaling is important in diverse physiological and pathological processes. The activity of rat phenol sulfotransferase (rSULT1A1), which is important for the metabolism of hormone and drug, is subjected to redox regulation. Two cysteines, Cys232 and Cys66, nanometer away from each other and from the enzyme active site were proposed to form disulfide bond to regulate the activity of rSULT1A1. A nano switch, composed of a flexible loop from amino acid residues 59-70, explained how this long distance interaction between two cysteines can be achieved. The enzyme properties were investigated through site-directed muatagnesis, circular dichroism, enzyme kinetics and homologous modeling of the rSULT1A1 structures. We proposed that the formation of disulfide bond between Cys232 and Cys66 induced conformational changes of sulfotransferase, then in turn affected its nucleotide binding and enzyme activity. This discovery was extended to understand the possible redox regulation of other sulfotransferases from different organisms. The redox switch can be created in other redox-insensitive sulfotransferases, such as human phenol sulfotransferase (hSULT1A1) and human alcohol sulfotransferase (hSULT2A1), to produce mutant enzymes with redox regulation capacity. This study strongly suggested that redox regulation of drug and hormone metabolism can be significantly varied even though the sequence and structure of SULT1A1 of human and rat have a high degree of homology.


Assuntos
Arilsulfotransferase/química , Arilsulfotransferase/metabolismo , Cisteína/química , Animais , Arilsulfotransferase/genética , Dicroísmo Circular , Dissulfetos , Humanos , Modelos Moleculares , Mutagênese Sítio-Dirigida , Oxirredução , Conformação Proteica , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Compostos de Sulfidrila/química , Sulfotransferases/química , Sulfotransferases/genética , Sulfotransferases/metabolismo
4.
Biosens Bioelectron ; 35(1): 456-460, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22425237

RESUMO

This work presents miniaturized CMOS (complementary metal oxide semiconductor) sensors for non-faradic impedimetric detection of AIV (avian influenza virus) oligonucleotides. The signal-to-noise ratio is significantly improved by monolithic sensor integration to reduce the effect of parasitic capacitances. The use of sub-µm interdigitated microelectrodes is also beneficial for promoting the signal coupling efficiency. Capacitance changes associated with surface modification, functionalization, and DNA hybridization were extracted from the measured frequency responses based on an equivalent-circuit model. Hybridization of the AIV H5 capture and target DNA probes produced a capacitance reduction of -13.2 ± 2.1% for target DNA concentrations from 1 fM to 10 fM, while a capacitance increase was observed when H5 target DNA was replaced with non-complementary H7 target DNA. With the demonstrated superior sensing capabilities, this miniaturized CMOS sensing platform shows great potential for label-free point-of-care biosensing applications.


Assuntos
Técnicas Biossensoriais/métodos , DNA Viral/análise , Vírus da Influenza A/isolamento & purificação , Influenza Aviária/virologia , Animais , Sequência de Bases , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/estatística & dados numéricos , Aves , Sondas de DNA/genética , DNA Viral/genética , Técnicas Eletroquímicas , Desenho de Equipamento , Fluorescência , Vírus da Influenza A/genética , Vírus da Influenza A/patogenicidade , Hibridização de Ácido Nucleico , Pontos Quânticos , Sensibilidade e Especificidade , Razão Sinal-Ruído
5.
Biosens Bioelectron ; 24(10): 3019-24, 2009 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-19362813

RESUMO

Enhanced surveillance of influenza requires rapid, robust, and inexpensive analytical techniques capable of providing a detailed analysis of influenza virus strains. Functionalized poly-crystalline silicon nanowire field-effect transistor (poly-SiNW FET) was demonstrated to achieve specific and ultrasensitive (at fM level) detection of high pathogenic strain virus (H5 and H7) DNA of avian influenza (AI) which is an important infectious disease and has an immediate need for surveillance. The poly-SiNW FET was prepared by a simple and low-cost method that is compatible with current commercial semiconductor process without expensive E-beam lithography tools for large-scale production. Specific electric changes were observed for AI virus DNA sensing when nanowire surface of poly-SiNW FET was modified with complementary captured DNA probe and target DNA (H5) at fM to pM range could be distinguished. With its excellent electric properties and potential for mass commercial production, poly-SiNW FET can be developed to become a portable biosensor for field use and point-of-care diagnoses.


Assuntos
Alphainfluenzavirus/isolamento & purificação , Técnicas Biossensoriais/instrumentação , DNA Viral/análise , Animais , Sequência de Bases , Técnicas Biossensoriais/métodos , Técnicas Biossensoriais/estatística & dados numéricos , Aves , Sondas de DNA/genética , DNA Viral/genética , Técnicas Eletroquímicas , Humanos , Influenza Aviária/diagnóstico , Alphainfluenzavirus/genética , Alphainfluenzavirus/patogenicidade , Nanofios , Vigilância da População/métodos , Sensibilidade e Especificidade , Silício , Transistores Eletrônicos
6.
Biosens Bioelectron ; 24(5): 1223-9, 2009 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-18760914

RESUMO

A simple and low-cost method to fabricate poly-silicon nanowire field effect transistor (poly-Si NW FET) for biosensing application was demonstrated. The poly-silicon nanowire (poly-Si NW) channel was fabricated by employing the poly-silicon (poly-Si) sidewall spacer technique, which approach was comparable with current commercial semiconductor process and forsaken expensive E-beam lithography tools. The electronic properties of the poly-Si NW FET in aqueous solution were found to be similar to those of single-crystal silicon nanowire field effect transistors reported in the literature. A model biotin and avidin/streptavidin sensing system was used to demonstrate the biosensing capacity of poly-Si NW FET. The changes of I(D)-V(G) curves were consistent with an n-type FET affected by a nearby negatively (streptavidin) and positively (avidin) charged molecules, respectively. Specific electric changes were observed for streptavidin and avidin sensing when nanowire surface of poly-Si NW FET was modified with biotin and streptavidin at sub pM to nM range could be distinguished. With its excellent electric properties and the potential for mass commercial production, poly-Si NW FET can be a very useful transducer for a variety of biosensing applications.


Assuntos
Técnicas Biossensoriais/instrumentação , Eletroquímica/instrumentação , Nanotecnologia/instrumentação , Nanotubos/química , Silício/química , Transistores Eletrônicos , Técnicas Biossensoriais/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Nanotecnologia/métodos , Nanotubos/ultraestrutura , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
7.
Chem Commun (Camb) ; (44): 5749-51, 2008 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-19009069

RESUMO

An unprecedented high sensitive sensing of neurotransmitter dopamine at fM level was demonstrated using a poly-crystalline silicon nanowire field-effect transistor (poly-SiNW FET) fabricated by employing a simple and low-cost poly-Si sidewall spacer technique, which was compatible with current commercial semiconductor processes for large-scale standard manufacture.


Assuntos
Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Dopamina/análise , Nanofios/química , Silício/química , Transistores Eletrônicos
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