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1.
ACS Omega ; 8(48): 46252-46260, 2023 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-38075742

RESUMO

Many countries have allowed farmers to feed ß-adrenergic receptor agonists, such as ractopamine (Rac), to animals to improve the quality of their meat. However, Rac consumption can cause health problems for humans; thus, detecting Rac in meat before its packaging is essential. Consequently, this study developed a simple and sensitive electrochemical sensor by modifying a glassy carbon electrode (GCE) with Nafion/silver nanoparticles (Nafion/AgNPs). When this electrochemical sensor is used to detect Rac, electrostatic interaction occurs between Nafion and Rac, and the AgNPs oxidize Rac; thus, the accumulation and electrochemical sensing of Rac are achieved. Differential pulse voltammetry indicated that the as-prepared Nafion/AgNP-GCE sensor exhibited suitable electrochemical sensing ability under optimum conditions (6.0 µL of 0.10% Nafion/AgNPs in a Britton-Robertson buffer solution with a pH of 1.8, an accumulation potential of -0.2 V, and a Rac accumulation duration of 300 s). Moreover, this sensor has an extremely low limit of detection and high sensitivity (1.60 × 10-3 ppm and 2.14 µA/ppm, respectively) in the Rac concentration range 7.50 × 10-3-1.00 ppm. The as-prepared sensor also exhibits satisfactory reproducibility and storage stability, with the corresponding relative standard deviations (RSDs) being 4.27% (n = 5) and 1.56% (n = 10), respectively. The proposed electrochemical sensor was successfully used to determine the Rac content in pig liver samples, with spiked recoveries of 95.2-101.8% and RSDs of 0.55-4.83% being achieved.

2.
BMC Mol Biol ; 8: 2, 2007 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-17212813

RESUMO

BACKGROUND: Despite of the fact that mammalian genomes are far more spacious than prokaryotic genomes, recent nucleotide sequencing data have revealed that many mammalian genes are arranged in a head-to-head orientation and separated by a small intergenic sequence. Extensive studies on some of these neighboring genes, in particular homologous gene pairs, have shown that these genes are often co-expressed in a symmetric manner and regulated by a shared promoter region. Here we report the identification of two non-homologous brain disease-related genes, with one coding for a serine protease inhibitor (SERPINI1) and the other for a programmed cell death-related gene (PDCD10), being tightly linked together by an asymmetric bidirectional promoter in an evolutionarily conserved fashion. This asymmetric bidirectional promoter, in cooperation with some cis-acting elements, is responsible for the co-regulation of the gene expression pattern as well as the tissue specificity of SERPINI1 and PDCD10. RESULTS: While SERPINI1 is predominantly expressed in normal brain and down-regulated in brain tumors, PDCD10 is ubiquitously expressed in all normal tissues but its gene transcription becomes aberrant in different types of cancers. By measuring the luciferase activity in various cell lysates, their 851-bp intergenic sequence was shown to be capable of driving the reporter gene expression in either direction. A 175-bp fragment from nt 1 to 175 in the vicinity of PDCD10 was further determined to function as a minimal bidirectional promoter. A critical regulatory fragment, from nt 176-473 outside the minimal promoter in the intergenic region, was identified to contain a strong repressive element for SERPINI1 and an enhancer for PDCD10. These cis-acting elements may exist to help coordinate the expression and regulation of the two flanking genes. CONCLUSION: For all non-homologous genes that have been described to be closely adjacent in the mammalian genomes, the intergenic region of the head-to-head PDCD10-SERPINI1 gene pair provides an interesting and informative example of a complex regulatory system that governs the expression of both genes not only through an asymmetric bidirectional promoter, but also through fine-tuned regulations with some cis-acting elements.


Assuntos
Proteínas Reguladoras de Apoptose/genética , Encefalopatias/genética , Sequência Conservada/genética , Evolução Molecular , Regulação da Expressão Gênica/genética , Proteínas de Membrana/genética , Neuropeptídeos/genética , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas/genética , Serpinas/genética , Sequência de Bases , Linhagem Celular , DNA Intergênico/genética , Deleção de Genes , Regulação Neoplásica da Expressão Gênica/genética , Saúde , Humanos , Dados de Sequência Molecular , Neoplasias/genética , Elementos Reguladores de Transcrição/genética , Neuroserpina
3.
J Virol ; 80(21): 10554-64, 2006 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16920820

RESUMO

The hepatitis B virus (HBV) X protein (pX) is implicated in hepatocarcinogenesis by an unknown mechanism. pX variants encoded by HBV genomes found integrated in genomic DNA from liver tumors of patients with hepatocellular carcinoma (HCC) generally lack amino acids 134 to 154. Since deregulation of mitogenic pathways is linked to oncogenic transformation, herein we define the pX region required for mitogenic pathway activation. A series of pX deletions was used to construct tetracycline-regulated pX-expressing cell lines. The activation of the mitogenic pathways by these pX deletions expressed in the constructed cell lines was measured by transient transreporter assays, effects on endogenous cyclin A expression, and apoptosis. Conditional expression of pX51-140 in AML12 clone 4 cell line activates the mitogenic pathways, induces endogenous cyclin A expression, and sensitizes cells to apoptosis, similar to wild-type (WT) pX. By contrast, pX1-115 is inactive, supporting the idea that amino acids 116 to 140 are required for mitogenic pathway activation. Moreover, this pX deletion analysis demonstrates that WT pX function is modulated by two regions spanning amino acids 1 to 78 and 141 to 154. The N-terminal X1-78, expressed via a retroviral vector in WT pX-expressing 4pX-1 cells, coimmunoprecipitates with WT pX, indicating this pX region participates in protein-protein interactions leading to pX oligomerization. Interestingly, pX1-78 interferes with WT pX in mediating mitogenic pathway activation, endogenous gene expression, and apoptosis. The C-terminal pX region spanning amino acids 141 to 154 decreases pX stability, determined by pulse-chase studies of WT pX and pX1-140, suggesting that increased stability of naturally occurring pX variants lacking amino acids 134 to 154 may play a role in HCC development.


Assuntos
Vírus da Hepatite B/fisiologia , Transativadores/química , Transativadores/fisiologia , Animais , Apoptose , Carcinoma Hepatocelular/etiologia , Carcinoma Hepatocelular/virologia , Linhagem Celular , Ciclina A/genética , Ciclina A/metabolismo , Expressão Gênica , Genes Virais , Vírus da Hepatite B/genética , Vírus da Hepatite B/patogenicidade , Humanos , Neoplasias Hepáticas/etiologia , Neoplasias Hepáticas/virologia , Camundongos , Mitógenos/química , Mitógenos/genética , Mitógenos/fisiologia , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Deleção de Sequência , Transdução de Sinais , Tetraciclina/farmacologia , Transativadores/genética , Transfecção , Proteínas Virais Reguladoras e Acessórias
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