Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 180
Filtrar
1.
Int J Rheum Dis ; 27(7): e15264, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-39016200

RESUMEN

OBJECTIVE: We employed two-sample Mendelian randomization (MR) to assess the genetic causal relationship between educational attainment (EA) and risk of five common connective tissue diseases (CTDs). METHODS: Educational attainment (self-reported at age ≥30 years) was obtained from a meta-analysis of years of schooling in 766 345 participants of European ancestry from genome-wide association studies (GWAS). A total of 1265 signals associated with EA were identified. Genetic data for five CTDs [rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), systemic sclerosis (SSc), polymyositis (PM), and dermatomyositis (DM)] were obtained from the FinnGen consortium. Two-sample MR analyses were performed separately for EA and the five CTDs. RESULTS: We found a negative causal relationship between EA and RA (ORIVW = 0.627, 95% CI = 0.537-0.732, p < .001), and SLE (ORIVW = 0.341, 95% CI = 0.123-0.944, p = .038). There were no genetic causal association between EA and SSc (ORIVW = 0.647, 95% CI = 0.351-1.195, p = .164), PM (ORIVW = 0.938, 95% CI = 0.320-2.746, p = .907), or DM (ORIVW = 0.754, 95% CI = 0.351-1.619, p = .468). None of the analyses revealed any horizontal pleiotropy or heterogeneity. CONCLUSION: Our findings indicated a potential causal association between EA and RA, SLE, emphasizing the need for further investigation and potential integration of EA into clinical practice to enhance treatment strategies.


Asunto(s)
Enfermedades del Tejido Conjuntivo , Escolaridad , Predisposición Genética a la Enfermedad , Estudio de Asociación del Genoma Completo , Análisis de la Aleatorización Mendeliana , Humanos , Factores de Riesgo , Enfermedades del Tejido Conjuntivo/genética , Enfermedades del Tejido Conjuntivo/epidemiología , Enfermedades del Tejido Conjuntivo/diagnóstico , Medición de Riesgo , Fenotipo , Masculino , Femenino , Lupus Eritematoso Sistémico/genética , Lupus Eritematoso Sistémico/epidemiología , Lupus Eritematoso Sistémico/diagnóstico , Adulto , Persona de Mediana Edad
2.
Food Chem ; 456: 139955, 2024 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-38852453

RESUMEN

The nanozyme-linked aptamer-sorbent assay (NLASA) is a rapid way to screen and characterize aptamer binding to targets. In this paper, a MnO2@AuNPs@aptamer (Apt) based NLASA coupled with colorimetric-SERS dual-mode for Staphylococcus aureus (S. aureus) detection is presented. Cu,Fe-CDs were used as the reducing agent to synthesize MnO2 and gold nanoparticles (AuNPs). Then, they were fabricated to obtain MnO2@AuNPs with oxidase (OXD)-like and SERS activities. The S. aureus aptamer was conjugated to MnO2@AuNPs and enhanced the OXD-like activity, which realized the specific capture of S. aureus in food matrices. In addition, S. aureus improves the oxidation of 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS) but inhibits 3,3',5,5'-tetramethylbenzidine (TMB) to generate Raman-active oxTMB with MnO2@AuNPs@Apt. This sensor was used for detections of S. aureus in a concentration ranged from 101 to 107 CFU/mL with a detection limit of 0.926 CFU/mL (colorimetric) and 1.561 CFU/mL (SERS), and the recovery is 85%-105% in real samples.

3.
Anal Methods ; 16(25): 4116-4123, 2024 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-38855960

RESUMEN

MicroRNAs (miRNAs) have been involved in many biological processes and are regarded as promising biomarkers. The short sequence, low abundance and highly homologous interference sequences greatly hinder the accurate detection of miRNAs. Here, a cascade branch migration-triggered strand displacement amplification (CBM-TSDA) strategy was developed for the first time for specific and sensitive detection of miRNA-155 (miR-155). In the presence of target miR-155, the CBM was initiated and two Y-shaped probes were eventually produced. Next, the Y-shaped probes were transformed into three-way junction (3WJ) structures and triggered the SDA to produce a large number of G-quadruplex (G4) structures. Finally, the increased fluorescence signal of G4/Thioflavin T (ThT) was used to quantify miR-155. Meanwhile, the colorimetric responses of the G4-hemin DNAzyme could be used as supplementary detection to obtain a dual-mode signal readout. This detection strategy showed high detection sensitivity, and the limit of detection was 0.28 pM in the fluorescence detection mode and 0.34 pM in the colorimetric detection mode. Notably, it showed high detection specificity, being able to discriminate the single-base mutations of the target with a high discrimination factor. The strategy also possessed excellent capacity for miR-155 detection in cell lysates and real human blood samples. The developed strategy provides a promising detection platform for miRNA, which may be applied to early clinical diagnosis.


Asunto(s)
Límite de Detección , MicroARNs , Técnicas de Amplificación de Ácido Nucleico , MicroARNs/sangre , MicroARNs/análisis , Humanos , Técnicas de Amplificación de Ácido Nucleico/métodos , G-Cuádruplex , Colorimetría/métodos , ADN Catalítico/química , ADN Catalítico/metabolismo , Técnicas Biosensibles/métodos , Espectrometría de Fluorescencia/métodos
4.
Plant Physiol ; 2024 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-38717740

RESUMEN

The circadian system plays a pivotal role in facilitating the ability of crop plants to respond and adapt to fluctuations in their immediate environment effectively. Despite the increasing comprehension of PSEUDO-RESPONSE REGULATORs (PRRs) and their involvement in the regulation of diverse biological processes, including circadian rhythms, photoperiodic control of flowering, and responses to abiotic stress, the transcriptional networks associated with these factors in soybean (Glycine max (L.) Merr.) remain incompletely characterized. In this study, we provide empirical evidence highlighting the significance of GmPRR3b as a crucial mediator in regulating the circadian clock, drought stress response, and abscisic acid (ABA) signaling pathway in soybeans. A comprehensive analysis of DNA affinity purification sequencing and transcriptome data identified 795 putative target genes directly regulated by GmPRR3b. Among them, a total of 570 exhibited a significant correlation with the response to drought, and eight genes were involved in both the biosynthesis and signaling pathways of ABA. Notably, GmPRR3b played a pivotal role in the negative regulation of the drought response in soybeans by suppressing the expression of abscisic acid responsive element-binding factor 3 (GmABF3). Additionally, the overexpression of GmABF3 exhibited an increased ability to tolerate drought conditions, and it also restored the hypersensitive phenotype of the GmPRR3b overexpressor. Consistently, studies on the manipulation of GmPRR3b gene expression and genome editing in plants revealed contrasting reactions to drought stress. The findings of our study collectively provide compelling evidence that emphasizes the significant contribution of the GmPRR3b-GmABF3 module in enhancing drought tolerance in soybean plants. Moreover, the transcriptional network of GmPRR3b provides valuable insights into the intricate interactions between this gene and the fundamental biological processes associated with plant adaptation to diverse environmental conditions.

5.
Talanta ; 277: 126265, 2024 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-38805947

RESUMEN

The pathogenic bacteria induced foodborne disease has been detrimental to public health worldwide. Herein, the peroxidase (POD)-like Fe3O4/MWCNTs@Mo-CDs (FMMC) nanozyme was applied for the detection of Escherichia coli (E. coli). The E. coli aptamer was conjugated with the surface of the FMMC, which effectively enhanced the POD-like activity attributing to the higher affinity to the substrate, and then specific capture of E. coli in food matrices, leading to the reduction of POD-like activity. Therefore, a robust and facile colorimetric aptasensor was developed for detecting E. coli with a wide linear range of 101-106 CFU/mL, low LOQ of 101 CFU/mL and LOD of 0.978 CFU/mL. The aptasensor demonstrated the satisfied selectivity for E. coli compared to the other strains. This method possessed the potential application for fast in situ screening of foodborne pathogens in food products.

6.
Medicine (Baltimore) ; 103(19): e38110, 2024 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-38728478

RESUMEN

RATIONAL: The Philadelphia chromosome (Ph) is seen in most patients with chronic myeloid leukemia and some patients with acute lymphoblastic leukemia. However, Ph-positive acute myeloid leukemia (Ph + AML) is a rare entity with a poor prognosis and a short median survival period. To date, there have been few clinical reports on this disease. And the treatment regimen of this disease has not been uniformly determined. PATIENT CONCERNS: We report a case of a Ph + AML. A 32-year-old male who was admitted to our hospital with weakness for 2 months. DIAGNOSIS: Philadelphia chromosome-positive acute myeloid leukemia. INTERVENTIONS: The patient achieved complete remission by the administration of a tyrosine kinase inhibitor, combined with low-intensity chemotherapy and a B-cell lymphoma 2 inhibitor. Then, allogeneic hematopoietic stem cell transplantation (allo-HSCT) from his sister was successfully performed. OUTCOMES: The patient has been in a continuous remission state for 6 months after transplantation. LESSONS: We reported a rare Ph + AML case, successfully treated with allo-HSCT. This case provided strong support for treating Ph + AML with allo-HSCT.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas , Leucemia Mieloide Aguda , Cromosoma Filadelfia , Humanos , Masculino , Trasplante de Células Madre Hematopoyéticas/métodos , Adulto , Leucemia Mieloide Aguda/terapia , Leucemia Mieloide Aguda/genética , Trasplante Homólogo , Inducción de Remisión
7.
Cell Prolif ; : e13645, 2024 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-38601993

RESUMEN

The biogenesis of exosomes that mediate cell-to-cell communication by transporting numerous biomolecules to neighbouring cells is an essential cellular process. The interaction between the transmembrane protein syndecan-4 (SDC4) and cytosolic protein syntenin plays a key role in the biogenesis of exosomes. However, how the relatively weak binding of syntenin to SDC4 efficiently enables syntenin sorting for packaging into exosomes remains unclear. Here, we demonstrate for the first time that SDC4 can undergo liquid-liquid phase separation (LLPS) to form condensates both in vitro and in the cell membrane and that, the SDC4 cytoplasmic domain (SDC4-CD) is a key contributor to this process. The phase separation of SDC4 greatly enhances the recruitment of syntenin to the plasma membrane (PM) despite the weak SDC4-syntenin interaction, facilitating syntenin sorting for inclusion in exosomes. Interestingly, phosphorylation at the only serine (179) in the SDC4-CD (Ser179) disrupts SDC4 LLPS, and inhibited phosphorylation or dephosphorylation restores the SDC4 LLPS to promote its recruitment of syntenin to the PM and syntenin inclusion into exosomes. This research reveals a novel phosphorylation-regulated phase separation property of SDC4 in the PM through which SDC4 efficiently recruits cytosolic syntenin and facilitates the biogenesis of exosomes, providing potential intervention targets for exosome-mediated biomedical events.

8.
Microbes Infect ; 26(5-6): 105344, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38670218

RESUMEN

To generate a new murine model for virus, DC-SIGN gene in murine was humanized. In this study, we successfully generated a humanized C57BL/6N mouse model expressing human DC-SIGN (hDC-SIGN) using CRISPR/Cas9 technology, and evaluated its characters and susceptibility to virus. The humanized mice could survival as usual, and with normal physiological index just like the wild-type mice. Whereas, we found significant differences in the intestinal flora and metabolic profiles between wild-type mice and humanized mice. Following intranasal infection with SARS-CoV-2, hDC-SIGN mice exhibited significantly increased viral loads in the lungs and nasal turbinates, along with more severe lung damage. This phenomenon may be associated with differential lipid metabolism and Fcγ receptor-mediated phagocytosis in two mouse models. This study provides a useful tool for investigating the mechanisms of coronavirus infection and potential drug therapies against novel coronavirus.


Asunto(s)
COVID-19 , Moléculas de Adhesión Celular , Modelos Animales de Enfermedad , Lectinas Tipo C , Ratones Endogámicos C57BL , Receptores de Superficie Celular , SARS-CoV-2 , Animales , Moléculas de Adhesión Celular/metabolismo , Moléculas de Adhesión Celular/genética , COVID-19/virología , Humanos , Ratones , Receptores de Superficie Celular/metabolismo , Receptores de Superficie Celular/genética , Lectinas Tipo C/metabolismo , Lectinas Tipo C/genética , Susceptibilidad a Enfermedades , Pulmón/virología , Carga Viral , Microbioma Gastrointestinal , Fagocitosis , Ratones Transgénicos , Receptores de IgG/genética , Receptores de IgG/metabolismo , Metabolismo de los Lípidos
9.
Front Med (Lausanne) ; 11: 1359461, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38495117

RESUMEN

Obstructive Sleep Apnea Syndrome (OSAS) affects 13-33% of males and 6-9% of females globally and poses significant treatment challenges, including poor adherence to Continuous Positive Airway Pressure (CPAP) and residual excessive sleepiness (RES). This review aims to elucidate the emerging interest in pharmacological treatments for OSAS, focusing on recent advancements in this area. A thorough analysis of extensive clinical trials involving various drugs, including selective dopamine reuptake inhibitors, selective norepinephrine inhibitors, combined antimuscarinic agents, and orexin agonists, was conducted. These trials focused on ameliorating respiratory metrics and enhancing sleep quality in individuals affected by OSAS. The studied pharmacological agents showed potential in improving primary outcomes, notably the apnea-hypopnea index (AHI) and the Epworth sleepiness scale (ESS). These improvements suggest enhanced sleep quality and symptom management in OSAS patients. With a deeper understanding of OSAS, pharmacological interventions are emerging as a promising direction for its effective management. This review provides a comprehensive overview of the current state of drug research in OSAS, highlighting the potential of these treatments in addressing the disorder's complex challenges.

10.
Neurochem Int ; 176: 105727, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38555055

RESUMEN

Temporal lobe epilepsy (TLE), the most common type of drug-resistant epilepsy, severely affects quality of life. However, the underlying mechanism of TLE remains unclear and deserves further exploration. Sorbs2, a key synaptic regulatory protein, plays an important role in the regulation of synaptic transmission in the mammalian brain. In this study, we aimed to investigate the expression pattern of Sorbs2 in a kainic acid (KA)-induced TLE mouse model and in patients with TLE to further determine whether Sorbs2 is involved in seizure activity and to explore the potential mechanism by which Sorbs2 affects seizures in this TLE mouse model. First, we found that the expression of Sorbs2 was obviously increased in the hippocampus and cortex of a TLE mouse model and in the temporal cortex of TLE patients, indicating an abnormal expression pattern of Sorbs2 in TLE. Importantly, subsequent behavioral analyses and local field potential (LFP) analyses of a TLE mouse model demonstrated that the downregulation of hippocampal Sorbs2 could prolong the latency to spontaneous recurrent seizures (SRSs) and protect against SRSs. We also found that the knockdown of Sorbs2 in the hippocampus could decrease excitatory synaptic transmission in pyramidal neurons (PNs) in the hippocampal CA1 region and reduce the expression levels of the AMPAR subunits GluA1 and GluA2. Thus, we speculated that Sorbs2 may promote epileptogenesis and the development of TLE by affecting AMPAR-mediated excitatory synaptic transmission in PNs in the CA1 region. Therefore, reducing the expression of hippocampal Sorbs2 could restrain epileptogenesis and the development of TLE.


Asunto(s)
Epilepsia del Lóbulo Temporal , Proteínas de Unión al ARN , Receptores AMPA , Convulsiones , Transmisión Sináptica , Animales , Femenino , Humanos , Masculino , Ratones , Epilepsia del Lóbulo Temporal/metabolismo , Epilepsia del Lóbulo Temporal/inducido químicamente , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/metabolismo , Hipocampo/efectos de los fármacos , Ácido Kaínico/farmacología , Ácido Kaínico/toxicidad , Ratones Endogámicos C57BL , Receptores AMPA/metabolismo , Convulsiones/metabolismo , Convulsiones/inducido químicamente , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/genética , Transmisión Sináptica/fisiología , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo
11.
Planta ; 259(4): 76, 2024 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-38418674

RESUMEN

MAIN CONCLUSION: Investigation the expression patterns of GmPT genes in response to various abiotic stresses and overexpression of GmPT11 in soybean hairy roots and Arabidopsis exhibited hypersensitivity to salt stress. Soybean is considered to be one of the significant oil crops globally, as it offers a diverse range of essential nutrients that contribute to human health. Salt stress seriously affects the yield of soybean through negative impacts on the growth, nodulation, reproduction, and other agronomy traits. The phosphate transporters 1(PHT1) subfamily, which is a part of the PHTs family in plants, is primarily found in the cell membrane and responsible for the uptake and transport of phosphorus. However, the role of GmPT (GmPT1-GmPT14) genes in response to salt stress has not been comprehensively studied. Here, we conducted a systematic analysis to ascertain the distribution and genomic duplications of GmPT genes, as well as their expression patterns in response to various abiotic stresses. Promoter analysis of GmPT genes revealed that six stress-related cis-elements were enriched in these genes. The overexpression of GmPT11 in soybean hairy roots and Arabidopsis exhibited hypersensitivity to salt stress, while no significant change was observed under low phosphate treatment, suggesting a crucial role in the response to salt stress. These findings provide novel insights into enhancing plant tolerance to salt stress.


Asunto(s)
Arabidopsis , Glycine max , Humanos , Glycine max/genética , Arabidopsis/genética , Estrés Fisiológico/genética , Estrés Salino/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Plantas Modificadas Genéticamente/genética
12.
J Craniofac Surg ; 2024 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-38408325

RESUMEN

OBJECTIVES: To observe the clinical effect of recombinant human alkaline fibroblast growth factor (rh-bFGF) combined with collagen sponge in the treatment of maxillofacial deepⅡ degree burn. METHODS: From January 2019 to January 2022, 96 patients with maxillofacial deep Ⅱ degree burns were randomly divided into a control group (N=48) and an observation group (N=48). The observation group was treated with rh-bFGF and collagen sponge after debridement, whereas the control group was treated with silver sulfadiazine ointment after debridement. The healing rate and healing time of the wounds were observed, interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-10, epidermal growth factor (EGF), endothelial growth factor growth factor (VEGF), and metalloproteinase tissue inhibitor 1 (TIMP-1) were measured. Vancouver Scar Scale (VSS) was used to evaluate the local scar at 6 months after wound healing in both groups. RESULTS: On the 10th, 14th, and 21st day of treatment, the wound healing rate in the observation group was higher than that in the control group (P<0.05), the wound healing time in the observation group was lower than that in the control group (P<0.05), and on the 14th day of treatment, the levels of TNF-α and IL-6 in the observation group were lower than those in the control group (P<0.05). The levels of IL-10 in the observation group were higher than those in the control group (P<0.05). The levels of EGF, VEGF, and TIMP-1 in the observation group were higher than those in the control group (P<0.05), and the scores of VSS in the observation group were lower than those in the control group (P<0.05). CONCLUSIONS: Rh-bFGF combined with collagen sponge can decrease the levels of TNF-α and IL-6 and increase the levels of IL-10, which can control the inflammation effectively, at the same time, it can increase the level of EGF, VEGF, and TIMP-1, promote wound healing, and reduce scar hyperplasia. The treatment protocol is simple, safe, effective, and suitable for clinical application.

13.
Analyst ; 149(4): 1002-1021, 2024 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-38204433

RESUMEN

The abnormal expression of human DNA methyltransferases (DNMTs) is closely related with the occurrence and development of a wide range of human cancers. DNA (cytosine-5)-methyltransferase-1 (DNMT1) is the most abundant human DNA methyltransferase and is mainly responsible for genomic DNA methylation patterns. Abnormal expression of DNMT1 has been found in many kinds of tumors, and DNMT1 has become a valuable target for the diagnosis and drug therapy of diseases. Nowadays, DNMT1 has been found to be involved in multiple cancers such as pancreatic cancer, breast cancer, bladder cancer, lung cancer, gastric cancer and other cancers. In order to achieve early diagnosis and for scientific research, various analytical methods have been developed for qualitative or quantitative detection of low-abundance DNMT1 in biological samples and human tumor cells. Herein, we provide a brief explication of the research progress of DNMT1 involved in various cancer types. In addition, this review focuses on the types, principles, and applications of DNMT1 detection methods, and discusses the challenges and potential future directions of DNMT1 detection.


Asunto(s)
Neoplasias de la Mama , ADN (Citosina-5-)-Metiltransferasas , Humanos , Femenino , ADN (Citosina-5-)-Metiltransferasas/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , ADN (Citosina-5-)-Metiltransferasa 1/metabolismo , Metilación de ADN , Neoplasias de la Mama/genética , ADN/metabolismo , Biomarcadores/metabolismo
14.
Chem Commun (Camb) ; 60(9): 1168-1171, 2024 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-38193242

RESUMEN

We report an electrochemical device for portable on-site detection of gaseous CH3I based on PVIm-F for the first time. The device achieves detection of gaseous CH3I with a significant selectivity and a low detection limit (0.474 ppb) in 20 min at 50 °C and 50% relative humidity, which is of great significance for achieving real-time on-site monitoring of radioactive hazardous environments.

15.
Talanta ; 269: 125456, 2024 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-38061202

RESUMEN

DNA methylation plays an important role in epigenetic modification. DNA methyltransferase (DNMT) is essential in the DNA methylation process, and its abnormal expression is closely related to cancer. In this study, we propose a novel biosensor platform (DS-GlaI-EXPAR) that combines hemi-methylated double-stranded DNA (dsDNA) as the substrate for DNMT1 with GlaI-assisted isothermal exponential amplification reaction (EXPAR) for rapid, simple, and sensitive detection of DNMT1 activity. The hemi-methylated dsDNA is fully methylated by DNMT1, and GlaI recognizes and cleaves the fully methylated sequence, generating terminal fragments that trigger EXPAR for efficient signal amplification. Whereas hemi-methylated dsDNA without DNMT1 will keep intact and cannot initiate EXPAR. DNMT1 activity can therefore be sensitively quantified by the real-time fluorescence signal of the DS-GlaI-EXPAR platform. The high-efficiency amplification of EXPAR and the recognition of GlaI enable the platform to overcome the inherent cumbersome and time-consuming shortcomings of traditional methods while meeting specificity and sensitivity. This DS-GlaI-EXPAR platform offers an impressively low limit of detection of 0.86 pg/µL and the entire detection process can be completed in a short time of 2.5 h in a single tube. Furthermore, DNMT1 activity detected by this platform in MCF-7 cells was significantly higher than that of HEK293 cells, and the inhibition of Apt. #9 was verified. This DNMT1 activity detection platform is very convenient and effective for the discovery of inhibitors and early cancer diagnosis.


Asunto(s)
ADN , Neoplasias , Humanos , Fluorescencia , Células HEK293 , Metilasas de Modificación del ADN , Técnicas de Amplificación de Ácido Nucleico/métodos , Metilación de ADN
16.
Phytopathology ; 114(2): 474-483, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-37589413

RESUMEN

Brassica yellows virus (BrYV) is an economically important virus on cruciferous species. In this study, a one-pot reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay coupled with the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system was developed for the detection of BrYV. The limit of detection of this method reached 32.8 copies of the BrYV ORF5, which is 100-fold more sensitive than the RT-LAMP method. Moreover, there was no cross-reactivity with other rapeseed-infecting RNA viruses or poleroviruses. We dried the CRISPR/Cas12a reagent in a trehalose and pullulan mixture to retain its efficacy at the RT-LAMP temperature of 63°C in order to allow portable BrYV detection in a water bath. The entire process can be performed in about 1 h, and a positive result can be rapidly and conveniently detected using a handheld UV lamp. In the field, the RT-LAMP-CRISPR/Cas12a assay was accurate and had higher sensitivity than RT-LAMP and reverse transcription-polymerase chain reaction assays. The novel RT-LAMP-CRISPR/Cas12a assay allows convenient, portable, rapid, low-cost, highly sensitive, and specific detection of BrYV and has great potential for on-site monitoring of BrYV.


Asunto(s)
Brassica , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Transcripción Reversa , Sistemas CRISPR-Cas , Enfermedades de las Plantas
17.
ACS Sens ; 9(1): 62-72, 2024 01 26.
Artículo en Inglés | MEDLINE | ID: mdl-38126108

RESUMEN

Rapid and simple nucleic acid detection is significant for disease diagnosis and pathogen screening, especially under specific conditions. However, achieving highly sensitive and specific nucleic acid detection to meet the time and equipment demand remains technologically challenging. In this study, we proposed a magnetic separation enhanced colorimetry biosensor based on a toehold-containing three-way junction (TWJ) induced multiple isothermal exponential amplification and the CRISPR/Cas14a (C-TEC) biosensor. The TWJ template was designed as a Y-X-Y structure. In the presence of the target, the formation of toehold-containing TWJ complex induced primer extension, leading to the generation of amplified single-stranded DNA; this amplified DNA could then bind to either the free TWJ template for EXPAR reaction or the toehold of the TWJ complex for toehold-mediated strand displacement, thereby enabling the recycling of the target. The amplification products could trigger CRISPR/Cas14a for efficient trans-cleavage and release the magnetically bound gold nanoparticle probes for colorimetry detection. Using Mycobacterium tuberculosis 16S rDNA as the target, the proposed C-TEC could detect 16S rDNA down to 50 fM by the naked eye and 20.71 fM by UV-vis detector at 520 nm within 90 min under optimal conditions. We successfully applied this biosensor to clinical isolates of Mycobacterium tuberculosis. In addition, the C-TEC biosensor also showed feasibility for the detection of RNA viruses. In conclusion, the proposed C-TEC is a convenient, fast, and versatile platform for visual detection of pathogen DNA/RNA and has potential clinical applications.


Asunto(s)
Nanopartículas del Metal , Mycobacterium tuberculosis , Mycobacterium tuberculosis/genética , Oro/química , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Nanopartículas del Metal/química , ADN Ribosómico , Fenómenos Magnéticos
18.
J Pharm Biomed Anal ; 239: 115922, 2024 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-38134706

RESUMEN

The amorphous Cu-containing phosphomolybdate (Cu-PTs) composite with high peroxidase (POD)-like activity at neutral conditions was explored as biosensors for raloxifene (RAF) detection. The strong attraction between negatively charged Cu-PTs and positively charged substrates 3,3',5,5'-tetramethylbenzidine (TMB), as well as the acceleration of the conversion of active Cu+/Cu2+ by the Cu/W bimetallic redox couples were demonstrated to play significant roles in POD-like activity in physiological environment. When RAF is presence, it can bind to the surface of Cu-PTs and changes the chemical signal on the material surface, leading to the decreased POD-like activity. Based on this, a colorimetric method was established for the sensitive assay of RAF with a lower limit of detection (LOD) of 0.025 mg/L and good recovery from 90.13% to 108.9%. This work paves a new way to design a POD-like colorimetric protocol for tracing RAF in pharmaceutical products and environmental samples.


Asunto(s)
Técnicas Biosensibles , Clorhidrato de Raloxifeno , Colorimetría/métodos , Peroxidasa , Peroxidasas , Técnicas Biosensibles/métodos , Peróxido de Hidrógeno
19.
Front Genet ; 14: 1231682, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37779912

RESUMEN

Introduction: Neuropathic pain remains a prevalent and challenging condition to treat, with current therapies often providing inadequate relief. Ozone therapy has emerged as a promising treatment option; however, its mechanisms of action in neuropathic pain remain poorly understood. Methods: In this study, we investigated the effects of ozone treatment on gene expression and metabolite levels in the brainstem and hypothalamus of a rat model, using a combined transcriptomic and metabolomic approach. Results: Our findings revealed significant alterations in key genes, including DCST1 and AIF1L, and metabolites such as Aconitic acid, L-Glutamic acid, UDP-glucose, and Tyrosine. These changes suggest a complex interplay of molecular pathways and region-specific mechanisms underlying the analgesic effects of ozone therapy. Discussion: Our study provides insights into the molecular targets of ozone treatment for neuropathic pain, laying the groundwork for future research on validating these targets and developing novel therapeutic strategies.

20.
Anal Biochem ; 680: 115313, 2023 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-37678583

RESUMEN

As newly developed synthetic enzymes with exceptional catalytic capabilities and outstanding stability, nanozymes have drawn considerable interest in the realm of sensing. Using a simple hydrothermal process, iodine and copper-doped carbon dots (Cu,I-CDs) with simulated enzymes were fabricated in the current investigation. Cu,I-CDs demonstrate peroxidase-mimicking function together with high catalytic effectiveness due to aforementioned features. This led to generation of a colorimetric sensor for quick and accurate quantitative assessment of morphine (MOR). The outcomes showed the method's usefulness for the colorimetric detection of MOR. The linear range for MOR detection is 0.25-25 µg/mL having a reduced detection limit of 64 ng/mL. This sensor's successful use in the analysis of MOR in biological material is more noteworthy.


Asunto(s)
Colorimetría , Cobre , Carbono , Catálisis , Derivados de la Morfina
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...