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Family environment, emotion regulation and biological sensitivity have been shown to be associated with adolescents' externalizing problem behaviours. However, findings regarding respiratory sinus arrhythmia (RSA) reactivity are mixed and sometimes contradictory. This study aims to clarify the roles of RSA reactivity and anger regulation in the relationship between negative family expressiveness (NFE) and adolescents' externalizing behaviour by measuring RSA reactivity during the Parent-Adolescent Interaction Task (PAIT), designed to simulate a naturalistic negative family environment. In this study, 125 Chinese adolescents (M = 13.95 years, SD = 0.95; 48% male) completed questionnaires assessing negative family expressiveness, anger regulation and externalizing problems. Additionally, we collected electrocardiogram and respiration data during both the resting period and a 10-min PAIT. Results showed that anger regulation mediated the relationship between NFE and externalizing problem behaviours. Moreover, adolescents' RSA reactivity moderated this mediation effect, even after controlling for baseline RSA. Greater RSA suppression potentially indicated greater susceptibility, with the relationship between NFE and anger regulation being more pronounced in adolescents with greater RSA suppression compared to those with lesser RSA suppression. These findings highlight the importance of considering physiological systems, especially within the context of adverse family environments, when studying the relationships with externalizing problems.
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The resected pâ ¢A-N2 non-small-cell lung cancer (NSCLC) patients who could benefit from postoperative radiotherapy (PORT) are not well-defined. The study explored the role of PORT on EGFR mutant and wild-type NSCLC patients. We retrospectively searched for resected pIIIA-N2 lung adenocarcinoma patients who underwent EGFR mutation testing. 80 patients with EGFR wild-type and 85 patients with EGFR mutation were included. 62 patients received PORT. In overall population, the median disease-free survival (DFS) was improved in PORT arm compared to non-PORT arm (22.9 vs. 16.1 months; p = 0.036), along with higher 2-year locoregional recurrence-free survival (LRFS) rate (88.3% vs. 69.3%; p = 0.004). In EGFR wild-type patients, PORT was associated with a longer median DFS (23.3 vs. 17.2 months; p = 0.044), and a higher 2-year LRFS rate (86.8% vs. 61.9%; p = 0.012). In EGFR mutant patients, PORT was not significantly correlated with improved survival outcomes. EGFR wild-type may a biomarker to identify the cohort that benefits from PORT.
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The rapid and sensitive detection of Escherichia/Shigella genera is crucial for human disease and health. This study introduces a novel series of piezoelectric quartz crystal (SPQC) sensors for detecting Escherichia/Shigella genera. In this innovative biosensor, we propose a new target and novel method for synthesizing long-range DNA. The method relies on the amplification of two DNA probes, referred to as H and P amplification (HPA), resulting in the products of long-range DNA named Sn. The new target was screened from the 16S rRNA gene and utilized as a biomarker. The SPQC sensor operates as follows: the Capture probe is modified on the electrodes. In the presence of a Displace probe and target, the Capture can form a complex with the Displace probe. The resulting complex hybridizes with Sn, bridging the gap between the electrodes. Finally, silver wires are deposited between the electrodes using Sn as a template. This process results in a sensitive response from the SPQC. The detection limit of the SPQC sensor is 1 CFU/mL, and the detection time is within 2 h. This sensor would be of great benefit for food safety monitoring and clinical diagnosis.
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Técnicas Biossensoriais , Escherichia , Técnicas Biossensoriais/métodos , Escherichia/genética , DNA Bacteriano/análise , DNA Bacteriano/genética , RNA Ribossômico 16S/genética , Eletrodos , Quartzo/química , Limite de Detecção , Sondas de DNA/química , Humanos , Técnicas de Amplificação de Ácido Nucleico , Técnicas EletroquímicasRESUMO
The detection of DNA methylation at cytosine/guanine dinucleotide (CpG) islands in promoter regions of tumor suppressor genes has great potential for early cancer screening, diagnosis, and prognosis monitoring. Nevertheless, achieving accurate, sensitive, cost-effective, and quantitative detection of target methylated DNA remains challenging. Herein, we propose a novel piezoelectric sensor (series piezoelectric quartz crystal (SPQC)) based on transcription activator-like effectors (TALEs) for detecting DNA methylation of Ras association domain family 1 isoform A (RASSF1A) tumor suppressor genes (R-5mC). The sensor employs TALEs-Ni magnetic beads to specifically recognize and separate the R-5mC, thereby improving the detection selectivity. The TALEs-Ni magnetic beads-R-5mC complex is sheared by a nucleic acid enzyme (DNAzyme) to release the single-stranded DNA (ST). ST initiates a catalyzed hairpin assembly (CHA) reaction on the surface of the electrode, which in turn triggers the hybridization chain reaction (HCR) and silver staining for enhanced detection sensitivity. The strategy exhibits a linear response in the detection of R-5mC in the range of 1 fM to 1 nM with a detection limit of 0.79 fM. R-5mC as low as 0.01% can be detected, even in the presence of large numbers of unmethylated DNA. The detection of R-5mC in circulating cell-free DNA (cfDNA) derived from clinical plasma specimens of lung cancer patients yielded satisfactory results.
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Técnicas Biossensoriais , Metilação de DNA , Humanos , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Técnicas Eletroquímicas , Genes Supressores de Tumor , Limite de Detecção , EletrodosRESUMO
N6-methyladenosine (6mA) plays a pivotal role in diverse biological processes, including cancer, bacterial toxin secretion, and bacterial drug resistance. However, to date there has not been a selective, sensitive, and simple method for quantitative detection of 6mA at single base resolution. Herein, we present a series piezoelectric quartz crystal (SPQC) sensor based on the specific recognition of transcription-activator-like effectors (TALEs) for locus-specific detection of 6mA. Detection sensitivity is enhanced through the use of a hybridization chain reaction (HCR) in conjunction with silver staining. The limit of detection (LOD) of the sensor was 0.63 pM and can distinguish single base mismatches. We demonstrate the applicability of the sensor platform by quantitating 6mA DNA at a specific site in biological matrix. The SPQC sensor presented herein offers a promising platform for in-depth study of cancer, bacterial toxin secretion, and bacterial drug resistance.
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Adenina , Técnicas Biossensoriais , DNA , Adenina/análogos & derivados , Adenina/análise , Adenina/química , Adenina/metabolismo , DNA/química , DNA/análise , Técnicas Biossensoriais/métodos , Limite de Detecção , Humanos , Quartzo/químicaRESUMO
BACKGROUND: Gastric cancer (GC) is a common cancer type with both high incidence and mortality. Recent studies have revealed an important role of circRNA in the development of GC. However, more experiments are needed to reveal the precise molecular mechanisms of circRNA in GC development. METHODS: Bioinformatics analysis was conducted to predict the potential role of circ_PABPC1 in GC and the target proteins of circ_PABPC1. Quantitative RT-PCR, Western blot and immunohistochemistry assays were conducted to detect the levels of circ_PABPC1, NF-κB p65, NF-κB p65 (Ser536) and ILK. MTT, Edu staining, cell scratch-wound and trans-well assays were carried out to detect cell proliferation, migration and invasion. The interaction between ILK and circ_PABPC1 was confirmed by RNA immunoprecipitation (RIP), RNA pull-down and fluorescence in situ hybridization assays. Genetically modified GC cells were injected into mice to evaluate the tumor growth performance. RESULTS: This study found that the high expression of circ_PABPC1 was associated with a poor prognosis of GC. The up-regulation of circ_PABPC1 promoted the proliferation, migration and invasion of GC cells. Circ_PABPC1 bound to ILK protein, thereby preventing the degradation of ILK. ILK mediated the effect of circ_PABPC1 on GC cells through activating NF-κB. CONCLUSION: circ_PABPC1 promotes the malignancy of GC cells through binding to ILK to activate NF-κB signaling pathway.
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Movimento Celular , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , NF-kappa B , Proteínas Serina-Treonina Quinases , RNA Circular , Transdução de Sinais , Neoplasias Gástricas , Neoplasias Gástricas/patologia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/genética , Humanos , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , RNA Circular/genética , RNA Circular/metabolismo , Proliferação de Células/genética , Animais , Camundongos , NF-kappa B/metabolismo , NF-kappa B/genética , Movimento Celular/genética , Linhagem Celular Tumoral , Camundongos Nus , Masculino , Prognóstico , Feminino , Camundongos Endogâmicos BALB C , Invasividade Neoplásica , Pessoa de Meia-Idade , Fator de Transcrição RelA/metabolismo , Fator de Transcrição RelA/genéticaRESUMO
Pathogenic bacteria significantly contribute to elevated morbidity and mortality rates, highlighting the urgent need for early and precise detection. Currently, there is a paucity of effective broad-spectrum methods for detecting pathogenic bacteria. We have developed an innovative proton-responsive series piezoelectric quartz crystal (PR-SPQC) platform for the broad-spectrum identification of pathogenic bacteria. This was achieved by retrieving and aligning sequences from the NCBI GenBank database to identify and validate 16S rRNA oligonucleotide sequences that are signatures of pathogenic bacteria but absent in humans or fungi. The hyperbranched rolling circle amplification, activated exclusively by the screened target, exponentially generates protons that are detected by SPQC through a 2D polyaniline (PANI) film. The PR-SPQC platform demonstrates broad-spectrum capabilities in detecting pathogenic bacteria, with a detection limit of 2 CFU/mL within 90 min. Clinical testing of blood samples yielded satisfactory results. With its advantages in miniaturization, cost efficiency, and suitability for point-of-care testing, PR-SPQC has the potential to be extensively used for the rapid identification of diverse pathogenic bacteria within clinical practice and public health sectors.
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Bactérias , Técnicas de Amplificação de Ácido Nucleico , Prótons , Bactérias/isolamento & purificação , Bactérias/genética , Humanos , RNA Ribossômico 16S/genética , Quartzo/química , Limite de DetecçãoRESUMO
This study aimed to explore the biological functions and underlying mechanism of circRNA acetyl-CoA carboxylase alpha (circACACA) in colorectal cancer (CRC). The RNA and protein levels were detected by qRT-PCR and western blot assays. The malignant capacities of CRC cells were analyzed by cell counting kit-8 (CCK-8), colony formation, flow cytometry, and transwell assays. The target relationship between miR-193a/b-3p and circACACA/histone deacetylase 3 (HDAC3) was determined by luciferase reporter assay and RNA immunoprecipitation. The binding of HDAC3 to the p53 promoter was validated by chromatin immunoprecipitation (ChIP). CRC cell growth and lung metastasis were evaluated in nude mice in vivo. High expression of circACACA was found in CRC tissues and cells, which was closely associated with the advanced tumor, lymph node, metastasis (TNM) stage, metastasis, and low overall survival rate. circACACA downregulation effectively delayed CRC cell proliferation and metastasis, but triggered apoptosis via inactivating the mevalonic acid (MVA) pathway. However, circACACA overexpression resulted in the opposite effects. Mechanistically, circACACA enhanced HDAC3 expression through sponging miR-193a/b-3p, which activated the MVA pathway via inhibiting the acetylation and transcription of p53. Moreover, rescue experiments confirmed that miR-193a/b-3p inhibition reversed the inhibitory effect of circACACA deficiency on CRC growth and metastasis. Moreover, circACACA overexpression-mediated malignant phenotypes of CRC cells were abrogated by HDAC3 knockdown. circACACA promoted CRC progression via regulating the miR-193a/b-3p/HDAC3/p53 axis to activate the MVA pathway, providing evidence for circACACA as a promising therapeutic target for CRC.
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Neoplasias Colorretais , MicroRNAs , Animais , Camundongos , Carcinogênese/genética , Transformação Celular Neoplásica , Neoplasias Colorretais/patologia , Ácido Mevalônico , Camundongos Nus , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/genética , Proteína Supressora de Tumor p53/genéticaRESUMO
Vascular dementia (VaD) is the second most prevalent dementia, which is attributable to neurovascular dysfunction. Currently, no approved pharmaceuticals are available. Taohong Siwu decoction (TSD) is a traditional Chinese medicine prescription with powerful antiapoptosis and anti-inflammatory properties. In this study, a network pharmacology approach together with molecular docking validation was used to explore the probable mechanism of action of TSD against VaD. A total of 44 active components, 202 potential targets of components, and 3,613 VaD-related targets including 161 intersecting were obtained. The potential chemical components including kaempferol, baicalein, beta-carotene, luteolin, quercetin, and beta-sitosterol involved in the inflammatory response, oxidative stress, and apoptosis might have potential therapeutic effects on the treatment of VaD. The potential core targets including AKT1, CASP3, IL1ß, JUN, and TP53 associated with cell apoptosis and inflammatory might account for the essential therapeutic effects of TSD in VaD. The results indicated that TSD protected against VaD through multicomponent and multitarget modes. Though the detailed mechanism of action of various active ingredients needs to be further illustrated, TSD still showed a promising therapeutic agent for VaD due to its biological activity.
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Demência Vascular , Medicamentos de Ervas Chinesas , Humanos , Simulação de Acoplamento Molecular , Demência Vascular/tratamento farmacológico , Farmacologia em Rede , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Medicina Tradicional Chinesa/métodosRESUMO
Background: Cuproptosis is a novel type of regulated cell death and is reported to promote tumor occurrence and progression. However, whether a cuproptosis-related signature has an impact on hepatocellular carcinoma (HCC) is still unclear. Materials and methods: We analyzed the transcriptome data of HCC from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) database, and searched for tumor types with different cuproptosis patterns through consistent clustering of cuproptosis genes. We then constructed a Cuproptosis-Related Genes (CRGs)-based risk signature through LASSO COX regression, and further analyzed its impact on the prognosis, clinical characteristics, immune cell infiltration, and drug sensitivity of HCC. Results: We identified the expression changes of 10 cuproptosis-related genes in HCC, and all the patients can be divided into two subtypes with different prognosis by applying the consensus clustering algorithm. We then constructed a cuproptosis-related risk signature and identified five CRGs, which were highly correlated with prognosis and representative of this gene set, namely G6PD, PRR11, KIF20A, EZH2, and CDCA8. Patients in the low CRGs signature group had a favorable prognosis. We further validated the CRGs signature in ICGC cohorts and got consistent results. Besides, we also discovered that the CRGs signature was significantly associated with a variety of clinical characteristics, different immune landscapes and drug sensitivity. Moreover, we explored that the high CRGs signature group was more sensitive to immunotherapy. Conclusion: Our integrative analysis demonstrated the potential molecular signature and clinical applications of CRGs in HCC. The model based on CRGs can precisely predict the survival outcomes of HCC, and help better guide risk stratification and treatment strategy for HCC patients.
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Accurate and rapid detection of nucleic acid plays a vital role in the clinical treatment of tuberculosis caused by Mycobacterium tuberculosis (M.TB). However, false-negative and false-positive results caused by base mismatches could affect the detection accuracy. Inspired by the unique property of CRISPR/Cas9, we proposed a new MSPQC M.TB sensor based on the CRISPR/Cas9 system, which can distinguish single-base mismatches in 10 bases from the protospacer adjacent motif (PAM) region. In the proposed sensor, single-stranded DNA on Au interdigital electrodes was used as a capture probe for the target and an initiator for hybridization chain reaction (HCR). HCR was used to generate long double-stranded DNA (dsDNA), which could span the Au interdigital electrodes. CRISPR/Cas9 was used as recognition components to recognize capture/target dsDNA. When the target existed, the capture probe hybridized with the target to form dsDNA, which could be recognized and cut by CRISPR/Cas9. Thus, the DNA connection between electrodes was cut off and resulted in the MSPQC response. When no target existed, the capture probe remained single-stranded and could not be recognized and cut by CRISPR/Cas9. Therefore, DNA connection between electrodes was reserved. Moreover, silver staining technology was utilized to improve the sensitivity of detection. M.TB was detected by the proposed sensor using specific sequence fragments of 16S rRNA of M.TB as the target. The detection time was down to 2.3 h. The limit of detection (LOD) was 30 CFU/mL.
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Mycobacterium tuberculosis , Ácidos Nucleicos , Sistemas CRISPR-Cas/genética , DNA/genética , Mycobacterium tuberculosis/genética , RNA Ribossômico 16SRESUMO
Despite the accumulating evidence for increased risks for behavioral problems in left-behind adolescents in China, little research has explored their HPA axis functioning, which is hypothesized to play a central role in the association between early adversity and health. In the present study, we designed a longitudinal study to examine HPA axis function in left-behind adolescents and its mediating role in the association between family emotional expressiveness and internalizing problems. Participants were 81 adolescents (44 female; 37 male) aged 11-16 years. Salivary cortisol samples were collected six times a day for two consecutive days on regular school days. Negative family expressiveness (NFE) and internalizing problems were measured using self-report questionnaires. The results showed that NFE was negatively associated with diurnal cortisol, and diurnal cortisol was negatively associated with internalizing problems. Further analysis showed that diurnal cortisol secretion measured by AUC (area under the curve) mediated the association between NFE and internalizing problems. Our findings extended the existing literature about left-behind children via a psychoneuroendocrinological perspective, documenting the negative consequences of the family environment for youth health and development.
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Hidrocortisona , Sistema Hipotálamo-Hipofisário , Adolescente , Criança , Feminino , Humanos , Hidrocortisona/análise , Sistema Hipotálamo-Hipofisário/química , Estudos Longitudinais , Masculino , Sistema Hipófise-Suprarrenal/química , Saliva/químicaRESUMO
BACKGROUND AND AIM: Colorectal cancer (CRC) is one of the major health issues in the world. Circ_0000677 has been shown to be upregulated in CRC with unclarified function and mechanism. Methyltransferase-like 3 (METTL3) acts as a regulator for gene expression via the mechanism of RNA N6 -methyladenosine (m6 A) in different types of cancer, which is under the control of SUMO1-based SUMOylation. We aim to investigate their roles in CRC progression. METHODS: Quantitative real-time polymerase chain reaction and Western blot were used to detect the expressions of METTL3, circ_0000677, and ATP binding cassette subfamily c member 1(ABCC1) in CRC patients' tissues and cell lines. The functions of ABCC1 and circ_0000677 in CRC were studied by manipulating their level via knocking down or overexpression. RNA pull-down and RNA immunoprecipitation assays were performed to identify the specific binding of target genes. The biological function of SUMOylation of METTL3 was investigated in vivo by xenograft mice tumor model. RESULTS: METTL3, circ_0000677, and ABCC1 were upregulated in CRC patients' samples and cell lines. Circ_0000677 positively regulates CRC cell proliferation and drug resistance via affecting ABCC1 expression. METTL3 facilitated circ_0000677 level via m6 A modification. METTL3 was regulated by SUMO1-mediated SUMOylation in CRC. Mutation of METTL3-K459 could suppress tumor growth in vivo via regulating circ_0000677/ABCC1 axis. CONCLUSIONS: Overall, our study revealed that circ_0000677 and its downstream target ABCC1 were upregulated in CRC cells, induced by the METTL3-mediated m6 A modification of circ_0000677 and SUMO1-mediated SUMOylation of METTL3. This work provided a new strategy for the therapeutic treatment of CRC.
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Neoplasias Colorretais , Metiltransferases , Animais , Proliferação de Células/genética , Neoplasias Colorretais/patologia , Humanos , Metiltransferases/genética , Camundongos , Sumoilação/genéticaRESUMO
To investigate the function of lncRNA HOXD-AS1 in cervical squamous cell carcinoma (CESC) and the underlying mechanism. The expressions of HOXD-AS1 and FRRS1 were analyzed on the online software GEPIA based on CESC-related information in The Cancer Genome Atlas (TCGA). Cervical cancer cells (SiHa and Hela) were accordingly transfected with pCDNA3.1-HOXD-AS1, sh-HOXD-AS1, sh-FRRS1 or pCDNA3.1-ELF1. After cell transfection, CCK-8, EDU and flow cytometry were applied for measurement of cell vitality, quantity and apoptosis, respectively. The relationship between HOXD-AS1 and FRRS1 was predicted on the online software LncMap and further verified by RNA binding protein immunoprecipitation. Nude mice were injected with stabilized SiHa cells transfected with sh-HOXD-AS1 to assess the tumorigenic ability of HOXD-AS1 in vivo. Immunohistochemistry detected the expression of the proliferation marker Ki-67. The levels of HOXD-AS1, ELF1 and FRRS1 were measured in vivo and in vitro. HOXD-AS1 and FRRS1 were overexpressed in CESC. After transfection of sh-HOXD-AS1, sh-ELF1 or sh-FRRS1, the proliferation of SiHa and Hela cells was inhibited and their apoptosis was promoted; while HOXD-AS1 overexpression had opposite effects on CESC development. Co-transfection of sh-FRRS1 and pCDNA3.1-HOXD-AS1 could abolish the tumor suppressive effect of FRRS1 knockdown. HOXD-AS1 elevated the level of FRRS1 by binding ELF1. Furthermore, HOXD-AS1 contributed to the CESC growth in mouse models. HOXD-AS1 promotes CESC by up-regulating FRRS1 via ELF1.
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RNA Longo não Codificante , Neoplasias do Colo do Útero , Animais , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Células HeLa , Humanos , Camundongos , Camundongos Nus , Proteínas Nucleares/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
Diseases caused by bacteria pose great challenges to human health. The key to reduce disease transmission and mortality is to develop accurate and rapid methods for the detection and identification of bacteria. Herein, a rapid bacteria 16S rDNA electrochemical sensor based on target-triggered hairpin self-assembly and tripedal DNA walker (TD walker) amplification strategy was constructed. Specific variable region of 16S rDNA fragment of bacteria was used as biomarker. The target-triggered hairpin self-assembly strategy was used to prepare a TD walker. The hairpin DNA probes labeled with ferrocene (Fc) were designed and modified on surface of electrode. The "legs" of TD walker hybridized with three hairpin probes and opened their hairpin structures. Exo III enzyme recognised hybrid duplexes and selectively digest hairpin probes. The "legs" of TD walker was released and hybridized with the other three hairpin probes. In this way, the enzyme drived the walkers to walk along electrode interface, until hairpin DNA probes were all removed from the electrode, the Fc was far away from electrode interface. A significantly current reduction signal was obtained and bacteria were detected by recording this response. This strategy was low-cost and scalable, it could continuously recycle low-concentration targets, thus enhanced the detection sensitivity. As the proof-of-concept work, the electrochemical sensor was utilized as detector. The limit of detection (LOD) of detecting Staphylococcus aureus (S. aureus) was 20 CFU mL-1 and detection time was less than 3 h. It was expected to be widely used in clinical early diagnosis.
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Técnicas Biossensoriais , DNA Ribossômico , Técnicas Eletroquímicas , Ouro , Humanos , Limite de Detecção , Técnicas de Amplificação de Ácido Nucleico , Staphylococcus aureus/genéticaRESUMO
Rapid diagnosis of tuberculosis disease (TB) still remained a pressing need for TB control efforts all over the world. However, the existing detection approaches cannot satisfy demand of rapid detection of clinical Mycobacterium tuberculosis (M. tuberculosis) because of the long detection time and high cost. Herein, we proposed a new M. tuberculosis piezoelectric sensor based on AuNPs-mediated enzyme assisted signal amplification. A hairpin-shaped DNA duplex with a protrusion of the 3' end was designed. In the presence of specific 16 S rDNA fragment of M. tuberculosis, the hairpin probe was opened, which triggered the selective cleavage of hairpin probe by Exonuclease III (Exo III), resulting in the release of uncut DNA probe and target DNA. The released target DNA hybridized with another hairpin-shaped DNA duplex, and a new digestion cycle was started, thus generating large amounts of uncut DNA probes. The uncut DNA was pulled to the electrode surface by the hybridization with capture probe modified on the electrode. Subsequently detection probe labeled AuNPs was hybridized with uncut DNA and entered between the two electrodes. The AuNPs linked to hybridized detection probe were grown in the HAuCl4 and Nicotinamide adenine dinucleotide (NADH) solution and offered the conductive connection between the gaps of electrode. The changes were monitored by the piezoelectric sensor. The piezoelectric biosensor could achieve a detection of M. tuberculosis (102-108 CFU mL-1) within 3 h, the detection limit (LOD) was 30 CFU mL-1. The methodology could be transformed into different microbial targets, which is suitable for further development of small portable equipment and multifunctional detection.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Mycobacterium tuberculosis , DNA Ribossômico , Ouro , Mycobacterium tuberculosis/genéticaRESUMO
As everyone knows, bacterial infectious diseases are serious hazards to human health in the world. Despite performing many methods towards the bacterial pollution, containing many detection and sterilization techniques, there is still a lack of effective means. Herein, a novel copper modified titanium (Cu@TiO2) nanocomposites were resoundingly synthesized via the well-known sol-gel process, which revealed a significant antibacterial activity under the illumination of sunlight. The XRD, Raman spectroscopy and TEM images showed that the Cu@TiO2 nanocomposites with a globular shape are anatase phase, Moreover, low temperature physical adsorption test and UV- visible spectrum indicate Cu0.01 @TiO2 owns a supernal specific area (80 m²/g) and the high visible light absorbing ability. Furthermore, the novel Cu@TiO2 nanocomposites showed an unprecedented photocatalytic capacity towards Escherichia Coli (E. coli) bacteria. In vitro, Cu@TiO2 nanocomposites can kill almost 98.7% E. coli under 60 min simulated solar light irradiation than that of TiO 2(31.3 %). This study suggests that the Cu@TiO2 will be as a potential material for ameliorating antibiotic-resistant bacteria in food detection.
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Nanocompostos , Titânio , Catálise , Cobre/farmacologia , Escherichia coli , Humanos , Luz , Titânio/farmacologiaRESUMO
The early detection of bacteria is of critical importance in addressing serious public health problems. Here, an electrochemical biosensor for rapid detection of bacteria based on facile synthesis of silver wire across electrodes was constructed. High-variable region of 16S rRNA of bacteria was used as biomarker. Polymerase-free synthesis of silver wire was introduced into electrochemical signal transduction to improve the sensitivity of electrochemical detection. The construction biosensor of proposed method is as follows: Metastable hairpin probe H1 was modified on electrode surface, biomarker can open the stem-loop structure of H1 and activates HCR. The alternate opening of the stem-loop structure of H1 and H2-AuNPs finally results in the formation of long double-stranded DNA-RNA (HCR products) -AuNPs. The formed AuNPs modified HCR products was blown in one direction using N2 to across the electrode gap. Using this HCR products as template, the silver wire was formed between the electrodes by silver deposition, and resulted in sharp change in electrical parameters of electrode. As the proof-of-concept work, multichannel series piezoelectric quartz crystal (MSPQC) was utilized as detector. The detection of Staphylococcus aureus in the concentration range from 50 to 107 CFU/mL within 100 min was achieved. The detection limit was 50 CFU/mL. Escherichia coli, Salmonella enteritidis, Listeria innocua, Pseudomonas aeruginosa and Streptococcus pneumoniae did not interfere the detection results. This newly proposed electrochemical biosensor is simple, rapid and exhibit high signal-to-noise ratio, it has great potential for being applied in food safety monitoring and clinical diagnosis.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Bactérias , Técnicas Eletroquímicas , Eletrodos , Ouro , Limite de Detecção , Listeria , RNA Ribossômico 16S/genética , PrataRESUMO
Tuberculosis is one of the life-threatening infectious diseases caused by the obligate pathogenic bacterium Mycobacterium tuberculosis (M. tuberculosis). The current M. tuberculosis detection approaches cannot satisfy the requirement for early clinical diagnosis because of long detection time as well as low specificity. In our study, an electrochemical M. tuberculosis sensor was constructed by using specific fragment of 16S rDNA of M. tuberculosis H37Ra as target biomarker, peptide nucleic acid (PNA) as capture probe and highly conductive two-dimensional Ti3C2 MXenes as the signal amplified transduction material. After the hybridization between PNA and the specific fragment of 16S rDNA on the substrate of PNA-AuNPs nanogap network electrode, the target fragments were directly linked with conductive Ti3C2 MXenes by strong interactions between zirconium-cross-linked Ti3C2 MXenes and phosphate groups of the target fragments. The linking of Ti3C2 MXenes to the hybridized target fragments would bridge the gaps of the interrupted AuNPs in the nanogap network electrode and forming the conductive connection to cause the change in conductance between the electrodes. This conductance change could be used for M. tuberculosis detection. The limit of detection (LOD) of proposed method was 20 CFU mL-1, and detection time was 2 h. Proposed method would find potential application in rapid detection of M. tuberculosis.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Mycobacterium tuberculosis/isolamento & purificação , Titânio/química , Condutividade Elétrica , Eletrodos , Estrutura Molecular , Tamanho da Partícula , Fosfatos/química , Propriedades de Superfície , Zircônio/químicaRESUMO
Herein, we developed an ultrasensitive electrochemical biosensor for Pseudomonas aeruginosa detection based on a rolling circle amplification-assisted multipedal DNA walking strategy. The cascade signal amplification method can improve the sensitivity, showing promising applications in food security, environmental monitoring, and disease diagnosis.