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1.
Ther Adv Hematol ; 15: 20406207241256894, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38828002

RESUMEN

Background: Pediatric acute myeloid leukemia (AML) has poor prognosis and high rate of relapse and mortality, and exploration of new treatment options is still critically needed. Objectives: To summarize the outcome of our new treatment strategies for pediatric AML, which is characterized by dual induction and acute lymphoblastic leukemia (ALL) elements consolidation. Design: Retrospective, single-arm study. Methods: From July 2012 to December 2019, an intensive chemotherapy protocol was used for newly diagnosed children with AML, which contains dual induction, three courses of consolidations based on high-dose cytarabine, and two courses of consolidations composed of high-dose methotrexate, vincristine, asparaginase, and mercaptopurine (ALL-like elements). Blasts were monitored by bone marrow smears at intervals, and two lumbar punctures were performed during chemotherapy. We retrospectively analyzed the efficacy and safety of this study. The last follow-up was on 26 May 2023. Results: A total of 70 pediatric AMLs were included. The median age at diagnosis was 6.7 (0.5-16.0) years. The median initial WBC count was 23.74 × 109/L, 11 of whom ⩾100 × 109/L. After dual induction, there were 62 cases of complete remission (CR), 5 cases of partial remission, and 3 cases of nonremission. The CR rate was 88.57%. The median follow-up time was 5.8 (0.2-9.4) years, the 5-year overall survival was 78.2% ± 5%, the event-free survival (EFS) was 71.2% ± 5.6%, and the cumulative recurrence rate was 27.75%. The 5-year EFS of patients with initial WBC < 100 × 109/L (n = 59) and ⩾100 × 109/L (n = 11) were 76.4% ± 5.7% and 45.5% ± 15% (p = 0.013), respectively. A total of 650 hospital infections occurred. The main causes of infection were respiratory tract infection (26.92%), septicemia (18.46%), stomatitis (11.85%), and skin and soft-tissue infection (10.46%). Conclusion: This intensive treatment protocol with dual induction and ALL-like elements is effective and safe for childhood AML. Initial WBC ⩾ 100 × 109/L was the only independent risk factor in this cohort. Trial registration: It is a retrospective study, and no registration on ClinicalTrials.gov.

2.
Technol Health Care ; 32(S1): 299-312, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38759058

RESUMEN

BACKGROUND: Plane-wave imaging is widely employed in medical imaging due to its ultra-fast imaging speed. However, the image quality is compromised. Existing techniques to enhance image quality tend to sacrifice the imaging frame rate. OBJECTIVE: The study aims to reconstruct high-quality plane-wave images while maintaining the imaging frame rate. METHODS: The proposed method utilizes a U-Net-based generator incorporating a multi-scale convolution module in the encoder to extract information at different levels. Additionally, a Dynamic Criss-Cross Attention (DCCA) mechanism is proposed in the decoder of the U-Net-based generator to extract both local and global features of plane-wave images while avoiding interference caused by irrelevant regions. RESULTS: In the reconstruction of point targets, the experimental images achieved a reduction in Full Width at Half Maximum (FWHM) of 0.0499 mm, compared to the Coherent Plane-Wave Compounding (CPWC) method using 75-beam plane waves. For the reconstruction of cyst targets, the simulated image achieved a 3.78% improvement in Contrast Ratio (CR) compared to CPWC. CONCLUSIONS: The proposed model effectively addresses the issue of unclear lesion sites in plane-wave images.


Asunto(s)
Procesamiento de Imagen Asistido por Computador , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Algoritmos
3.
J Colloid Interface Sci ; 667: 136-146, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38636215

RESUMEN

Breaking through the limitations of lithium-ion transmission is imperative for high-power rechargeable batteries. As a promising anode material for fast-charging lithium-ion batteries (LIBs), niobium pentoxide (Nb2O5) has garnered considerable research attention due to its exceptional rate performance, stable lithium storage performance and high safety attributes. Nevertheless, the limited intrinsic conductivity of Nb2O5, coupled with its structural degradation during the cycling process, imposes constraints on its viability as a commercially viable electrode material. Herein, a ruthenium (Ru) doping method is employed to regulate the oxygen defects and the interlayer spacing of the tetragonal Nb2O5 (M-Nb2O5), offering superior reaction kinetics, higher stability for lithium storage sites and more unobstructed lithium-ion transport channels. Ru-doped Nb2O5 (RNO) manifests excellent electrochemical properties, including remarkable rate capacity (166 mAh/g at 80C), reversible capacity (246.98 mAh/g at 0.5C), improved initial Coulombic efficiency (95.77 % compared to 81.44 % of the pure sample) and cycling stability (maintaining a capacity of 113.5 mAh/g at 10C for 2,000 cycles). The enhancement mechanism of Ru doping on the structural stability and ion transport kinetics in tetragonal Nb2O5 is comprehensively elucidated through diverse electrochemical analyses and in-situ techniques.

4.
Technol Health Care ; 32(1): 31-53, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-37781821

RESUMEN

BACKGROUND: Pulse-inversion-based tissue harmonic imaging has been utilized for many years because it can effectively eliminate the harmonic leakage and produce low side-lobe. However, the pulse inversion method is sensitive to imaging object movements, which may result in motion artifacts. Spatial resolution and contrast were limited. OBJECTIVE: To improve ultrasound image quality by a new pulse-inversion-based tissue harmonic imaging technique. METHODS: Continuous wavelet transform is applied to investigate the correlation between mother wavelet and the received echoes from two opposite pulses. To get a better correlation, a novel mother wavelet named 'tissue wavelet' is designed based on the Khokhlov-Zabolotskaya- Kuznetsov (KZK) wave equation. Radio frequency data were obtained from open Ultrasonix SonixTouch imaging system. Experiments were carried on ultrasonic tissue phantom, human carotid artery and human liver. RESULTS: The average improvement of lateral spatial resolution is 49.52% compared to pulse-inversion-based tissue second-harmonic Imaging (PIHI). Contrast ratio (CR) and contrast-to-noise ratio (CNR) increased by 5.55 dB and 1.40 dB over PIHI. Tissue wavelet performs better than Mexh and Morl wavelet in lateral spatial resolution, CR, and CNR. CONCLUSION: The proposed technique effectively improves the imaging quality in lateral spatial resolution, CR, and CNR.


Asunto(s)
Hígado , Análisis de Ondículas , Humanos , Ultrasonografía/métodos , Hígado/diagnóstico por imagen , Movimiento (Física) , Movimiento , Fantasmas de Imagen
5.
Front Chem ; 11: 1253685, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37867995

RESUMEN

Detection of per- and polyfluoroalkyl substances (PFASs) is crucial in environmental mitigation and remediation of these persistent pollutants. We demonstrate that time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a viable technique to analyze and identify these substances at parts per trillion (ppt) level in real field samples without complicated sample preparation due to its superior surface sensitivity. Several representative PFAS compounds, such as perfluorooctanesulfonic acid (PFOS), perfluorobutanoic acid (PFBA), perfluoropentanoic acid (PFPeA), perfluoheptanoic acid (PFHpA), and perfluorononanoic acid (PFNA), and real-world groundwater samples collected from monitoring wells installed around at a municipal wastewater treatment plant located in Southern California were analyzed in this work. ToF-SIMS spectral comparison depicts sensitive identification of pseudo-molecular ions, characteristic of reference PFASs. Additionally, principal component analysis (PCA) shows clear discrimination among real samples and reference compounds. Our results show that characteristic molecular ion and fragments peaks can be used to identify PFASs. Furthermore, SIMS two-dimensional (2D) images directly exhibit the distribution of perfluorocarboxylic acid (PFCA) and PFOS in simulated mixtures and real wastewater samples. Such findings indicate that ToF-SIMS is useable to determine PFAS compounds in complex environmental water samples. In conclusion, ToF-SIMS provides simple sample preparation and high sensitivity in mass spectral imaging, offering an alternative solution for environmental forensic analysis of PFASs in wastewater in the future.

6.
Microorganisms ; 11(7)2023 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-37513013

RESUMEN

Diatoms contribute to carbon fixation in the oceans by photosynthesis and always form biofouling organized by extracellular polymeric substances (EPS) in the marine environment. Bacteria-produced quorum-sensing signal molecules N-acyl homoserine lactones (AHLs) were found to play an important role in the development of Cylindrotheca sp. in previous studies, but the EPS composition change was unclear. This study used the technology of alcian blue staining and scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and time-of-flight secondary ion mass spectrometry (ToF-SIMS) to directly observe the biofilm formation process. The results showed that AHLs promote the growth rates of diatoms and the EPS secretion of biofilm components. AHLs facilitated the diatom-biofilm formation by a forming process dependent on the length of carbon chains. AHLs increased the biofilm thickness and the fluorescence intensity and then altered the three-dimensional (3D) structures of the diatom-biofilm. In addition, the enhanced EPS content in the diatom-biofilm testified that AHLs aided biofilm formation. This study provides a collection of new experimental evidence of the interaction between bacteria and microalgae in fouling biofilms.

7.
Biomark Res ; 10(1): 68, 2022 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-36071474

RESUMEN

BACKGROUND: Neuroblastoma (NBL) is the most common extra-cranial solid tumour in childhood, with prognosis ranging from spontaneous remission to high risk for rapid and fatal progression. Despite existing therapy approaches, the 5-year event-free survival (EFS) for patients with advanced NBL remains below 30%, emphasizing urgent necessary for novel therapeutic strategies. Studies have shown that epigenetic disorders play an essential role in the pathogenesis of NBL. However, the function and mechanism of N7-methylguanosine (m7G) methyltransferase in NBL remains unknown. METHODS: The expression levels of m7G tRNA methyltransferase Methyltransferase-like 1 (METTL1) were analyzed by querying the Gene Expression Omnibus (GEO) database and further confirmed by immunohistochemistry (IHC) assay. Kaplan-Meier, univariate and multivariate cox hazard analysis were performed to reveal the prognostic role of METTL1. Cell function assays were performed to evaluate how METTL1 works in proliferation, apoptosis and migration in cell lines and xenograft mouse models. The role of METTL1 on mRNA translation activity of NBL cells was measured using puromycin intake assay and polysome profiling assay. The m7G modified tRNAs were identified by tRNA reduction and cleavage sequencing (TRAC-seq). Ribosome nascent-chain complex-bound mRNA sequencing (RNC-seq) was utilized to identify the variation of gene translation efficiency (TE). Analyzed the codon frequency decoded by m7G tRNA to clarify the translation regulation and mechanism of m7G modification in NBL. RESULTS: This study found that METTL1 were significantly up-regulated in advanced NBL, which acted as an independent risk factor and predicted poor prognosis. Further in NBL cell lines and BALB/c-nu female mice, we found METTL1 played a crucial role in promoting NBL progression. Furthermore, m7G profiling and translation analysis revealed downregulation of METTL1 would inhibit puromycin intake efficiency of NBL cells, indicating that METTL1 did count crucially in regulation of NBL cell translation. With all tRNAs with m7G modification identified in NBL cells, knockdown of METTL1 would significantly reduce the levels of both m7G modification and m7G tRNAs expressions. Result of RNC-seq shew there were 339 overlapped genes with impaired translation in NBL cells upon METTL1 knockdown. Further analysis revealed these genes contained higher frequency of codons decoded by m7G-modified tRNAs and were enriched in oncogenic pathways. CONCLUSION: This study revealed the critical role and mechanism of METTL1-mediated tRNA m7G modification in regulating NBL progression, providing new insights for developing therapeutic approaches for NBL patients.

8.
Biosens Bioelectron ; 202: 114000, 2022 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-35065478

RESUMEN

In current work, with elaborate designs of G-quadruplex containing "Y" junction structures, we demonstrate the construction of several new and label-free electrochemical logic gate operations (OR, AND, NOR, and NAND) by defining two distinct biomolecules, human 8-oxo-7,8-dihydroguanine DNA glycosylase 1 (hOGG1) and microRNA-141 (miRNA-141), as the inputs. The "Y" junction structures are immobilized onto the surface of gold electrode as the signal transduction platform. The presence of the input molecules with different combinations can alter the "Y" junction structures to disrupt the formation of the complete G-quadruplexes via 8-oxoG-site specific cleavage and miRNA-141-triggered displacement of the "Y" junctions. Subsequent association of hemin with the G-quadruplex sequences thus yields significant current variation outputs upon electrochemical reduction of hemin on the electrode, leading to the successful function of different logic operations without the involvement of labeling the DNA sequences with electro-active species. Featured with the advantages of multiple logic operations with distinct inputs and the label-free electrochemical format, such molecular logic gates can potentially provide promising opportunities for the development of simple and robust biological logic gates for various applications.


Asunto(s)
Técnicas Biosensibles , G-Cuádruplex , MicroARNs , Computadores Moleculares , Oro/química , Hemina , Humanos , Lógica
9.
Zhongguo Zhong Yao Za Zhi ; 46(9): 2237-2244, 2021 May.
Artículo en Chino | MEDLINE | ID: mdl-34047126

RESUMEN

In this study, HPLC-ESI-MS and HPLC methods were established to explore the differences in the main chemical components and content of Mori Cortex with(mulberry root bark) and without(Mori Cortex) the phellem layer from both qualitative and quantitative aspects. The HPLC-ESI-MS method was used for quality analysis in positive and negative ion modes, and 33 compounds were identified in mulberry root bark, 22 compounds in Mori Cortex, and 26 compounds in phellem layer; mulberry root bark and Mori Cortex shared 22 components, and mulberry root bark has 11 unique compounds; Mori Cortex and its phellem layer shared 15 components, while Mori Cortex has 7 unique compounds. HPLC method was used to simultaneously determine 7 major constituents, including mulberroside A, chlorogenic acid, dihydromorin, oxyresveratrol, moracin O, kuwanon G, and kuwanon H, and the developed method showed good linearity(r>0.998 9) within the concentration range and the recoveries varied from 99.88% to 103.0%, and the RSD was 1.7%-2.9%. The HPLC results showed that the contents of the 7 compounds have great differences in 13 batches samples, compared with mulberry root bark, the contents of mulberroside A, chlorogenic acid, dihydromorin and moracin O of Mori Cortex were increased, while the contents of oxyresveratrol, kuwanon G and kuwanon H were decreased after peeling process. These results can provide a basis for the rationality and quality control of Mori Cortex required to remove the phellem layer.


Asunto(s)
Medicamentos Herbarios Chinos , Morus , Cromatografía Líquida de Alta Presión , Espectrometría de Masas , Corteza de la Planta
10.
Food Chem ; 358: 129894, 2021 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-33933968

RESUMEN

Listeria monocytogenes is an important foodborne pathogen, which imposes great burdens on public health. The current methods for detecting L. monocytogene are limited in several ways such as time consuming and lab equipment dependent. In this study, we developed a new electrochemical assay to improve the efficacy. This assay allows us to generate numerous G-quadruplex sequences while loop-mediated isothermal amplification happens. Then, these G-quadruplex sequences form DNAzyme to produce a color change and an electrochemical signal by oxidizing tetramethylbenzidine. This assay could be finished in 2 h, which significantly reduced the detection time. Also, we confirmed the limit of detection of this assay at 6.8 CFU/mL according to 3σ criterion. Our assay shows good sensitivity to detect bacteria range from 52.5 to 5.25 × 104 CFU/mL. This assay's reliability was also confirmed by detecting artificially contaminated pork samples. Thus, we propose this electrochemical assay for rapid and sensitive detection of L. monocytogenes in food.


Asunto(s)
Técnicas Electroquímicas/métodos , Microbiología de Alimentos/métodos , Listeria monocytogenes/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos , Carne de Cerdo/microbiología , Animales , Bencidinas/química , ADN Catalítico/química , ADN Catalítico/metabolismo , G-Cuádruplex , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
11.
Exp Ther Med ; 20(6): 281, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33209125

RESUMEN

This study analyzed the infection of superficial mycosis and the relationship between the distribution characteristics of pathogenic fungi and age, time and sex in Northeast China in the past 10 years. We would like to provide a theoretical basis for the diagnosis and treatment of related diseases. From December 2008 to December 2018, 5,374 superficial mycoses from Northeast China were selected. The fungal species were identified by fungal microscopy, fungal culture, and species identification. Besides, the relationship between sex, age, time and the distribution of superficial mycosis and pathogenic fungi was analyzed. Among the 5,374 patients, the top three infections were tinea pedis (n=1,538, 28.62%), tinea cruris (n=1,018, 18.94%) and tinea corporis (n=938, 17.45%). The top three pathogens were Trichophyton rubrum (n=2,849, 48.65%), Trichophyton mentagrophytes (n=947, 16.14%) and Candida spp. (n=804, 13.70%). The main pathogenic fungi were dermatophytes. The age group with the highest incidence of tinea capitis was children (n=372, 6.92%). The highest incidence rate of tinea pedis was in 31-69-year adults (n=905, 16.84%); Malassezia mainly affects young people aged 15-30. Yeast and mold mostly invade the elderly patients >60 years old. The incidence of tinea cruris, tinea pedis and tinea corporis in male patients was higher than that in female patients. The incidence of onychomycosis in female patients was higher than that in male patients (P<0.05). The isolation rate of Candida, Mold, Microsporum canis, Malassezia and Sporothrix increased year by year, while that of Trichophyton rubrum, Trichophyton mentagrophyte, Trichophyton schoenleinii and Epidermophyton floccosum decreased. From December 2008 to December 2018, dermatophytes were the main pathogens of superficial mycosis in Northeast China. The distribution of disease species and pathogenic fungi varied in different gender, age and time.

12.
Epidemiol Infect ; 148: e102, 2020 05 08.
Artículo en Inglés | MEDLINE | ID: mdl-32381145

RESUMEN

HIV-1 drug resistance can compromise the effectiveness of antiretroviral therapy (ART). A survey of pretreatment HIV-1 drug resistance (PDR) was conducted in Lincang Prefecture of Yunnan Province. From 372 people living with HIV/AIDS initiating ART for the first time during 2017-2018, 322 pol sequences were obtained, of which 11 HIV-1 strain types were detected. CRF08_BC (70.2%, 226/322) was the predominant strain, followed by URF strains (10.6%, 34/322). Drug resistance mutations (DRMs) were detected among 34.2% (110/322) of the participants. E138A/G/K/R (14.3%, 46/322) and V179E/D/T (13.7%, 47/322) were the predominant DRMs. Specifically, E138 mutations commonly occurred in CRF08_BC (19.9%, 45/226). Among the DRMs detected, some independently conferred resistance, such as K65R (1.6%, 5/322), Y188C/F/L (0.9%, 3/322), K103N (0.6%, 2/322) and G190A (0.3%, 1/322), which conferred high-level resistance. The prevalence of PDR was 7.5% (95% CI: 4.6-10.3%) and the prevalence of non-nucleotide reverse transcriptase inhibitor (NNRTI) resistance was 5.0% (95% CI: 2.6-7.4%), which is below the threshold (⩾10%) of initiating a public health response. In conclusion, HIV-1 genetic diversity and an overall moderate level of PDR prevalence were found in western Yunnan. PDR surveillance should be continually performed to decide whether a public health response to NNRTI resistance should be initiated.


Asunto(s)
Fármacos Anti-VIH/farmacología , Farmacorresistencia Viral , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , VIH-1/efectos de los fármacos , Adolescente , Adulto , Anciano , Fármacos Anti-VIH/uso terapéutico , China/epidemiología , Femenino , Infecciones por VIH/epidemiología , VIH-1/genética , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Adulto Joven
13.
J Basic Microbiol ; 60(1): 37-46, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31696958

RESUMEN

Geobacter metallireducens GS15, a model of dissimilatory iron-reducing bacteria, is the key regulator in biogeochemical iron cycling. How the emerging contaminant microplastics involved in the iron cycling are driven by microbes on the microscale remains unknown. Hence, the influences of two typical microplastics, polybutylene terephthalate-hexane acid (PBAT) and polyvinyl chloride (PVC), were explored on the activity of G. metallireducens GS15 with ferrihydrite or ferric citrate as the respective electron acceptors. The results showed that the iron (II) contents in PBAT- and PVC-treatment groups were 16.79 and 6.81 mM, respectively, at the end of the experiment. Compared with the PBAT-treatment group, scanning electron microscopy and energy dispersive spectrometery revealed that merely a small amount of iron-containing products covered the surface of PVC. Moreover, PBAT and PVC could both retard the electroactivity of G. metallireducens GS15 at the beginning of microbial fuel cell operation. On the basis of the results above, microplastic PVC might exhibit potential inhibition of the iron cycling process driven by G. metallireducens GS15 with ferrihydrite as the terminal electron acceptor. This study extended our understanding of the influence of the microplastics PBAT and PVC on microbially mediated biogeochemical iron cycling. The findings might have an important implication on the biogeochemical elements cycling in the ecosystem with the involvement of emerging contaminant microplastics.


Asunto(s)
Contaminantes Ambientales/toxicidad , Compuestos Férricos/metabolismo , Geobacter/efectos de los fármacos , Microplásticos/toxicidad , Cloruro de Polivinilo/toxicidad , Electroquímica , Geobacter/metabolismo , Oxidación-Reducción , Poliésteres/toxicidad
14.
Biointerphases ; 14(5): 051004, 2019 09 25.
Artículo en Inglés | MEDLINE | ID: mdl-31554406

RESUMEN

In situ liquid time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a powerful technique to study surface characterization of living biofilms in hydrated conditions. However, ToF-SIMS data analysis is still a great challenge in complicated bacterial biofilms, because many interference peaks from the medium may result in inaccurate interpretation. In this study, two syntrophic Geobacter populations are investigated using in situ liquid ToF-SIMS to reveal the biofilm surface changes between them due to direct interspecies electron transfer. By comparing spectral principal component analysis (PCA) results of all peaks and selected peaks, the authors find that spectral peak overlay is an effective strategy to reduce the matrix effect in handling complex ToF-SIMS data. Additionally, the spectral PCA results of high intensity and high resolution data obtained from liquid ToF-SIMS are compared. Selected peaks, amino acid peaks, and water cluster peaks spectral PCA produce nice separation among samples in both high intensity and high resolution data sets. However, the high resolution data show better separation between coculture planktonic and coculture aggregates, confirming that the higher mass accuracy is useful in the analysis of microbial samples. In conclusion, the results show that peak selection is critical for acquiring effective microbial information and interpretation of syntrophic Geobacter using spectral data from in situ liquid ToF-SIMS.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Geobacter/fisiología , Espectrometría de Masa de Ion Secundario , Aminoácidos/metabolismo , Transporte de Electrón/fisiología , Análisis de Componente Principal
15.
Anal Biochem ; 553: 7-11, 2018 07 15.
Artículo en Inglés | MEDLINE | ID: mdl-29777679

RESUMEN

Listeria monocytogenes is an important foodborne pathogen, and it can cause severe diseases. Rapid detection of L. monocytogenes is crucial to control this pathogen. A simple and robust strategy based on the cascade of PCR and G-quadruplex DNAzyme catalyzed reaction was used to detect L. monocytogenes. In the presence of hemin and the aptamer formed during PCR, the catalytic horseradish peroxidase-mimicking G-quadruplex DNAzymes allow the colorimetric responses of target DNA from L. monocytogenes. This assay can detect genomic DNA of L. monocytogenes specifically with as low as 50 pg/reaction with the naked eye. Through 20 pork samples assay, visual detection assay had the same results as conventional detection methods, and had a good performance. This is a powerful demonstration of the ability of G-quadruplex DNAzyme to be used for PCR-based assay with significant advantages of high sensitivity, low cost and simple manipulation over existing approaches and offers the opportunity for application in pathogen detection.


Asunto(s)
ADN Catalítico/metabolismo , Microbiología de Alimentos , Listeria monocytogenes/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Transducción de Señal , Catálisis , Colorimetría/métodos , Peroxidasa de Rábano Silvestre/química
16.
Virol J ; 15(1): 62, 2018 04 04.
Artículo en Inglés | MEDLINE | ID: mdl-29615087

RESUMEN

BACKGROUND: Iris yellow spot virus (IYSV) is an Orthotospovirus that infects most Allium species. Very few approaches for specific detection of IYSV from infected plants are available to date. We report the development of a high-sensitive Luminex xMAP-based microsphere immunoassay (MIA) for specific detection of IYSV. RESULTS: The nucleocapsid (N) gene of IYSV was cloned and expressed in Escherichia coli to produce the His-tagged recombinant N protein. A panel of monoclonal antibodies (MAbs) against IYSV was generated by immunizing the mice with recombinant N protein. Five specific MAbs (16D9, 11C6, 7F4, 12C10, and 14H12) were identified and used for developing the Luminex xMAP-based MIA systems along with a polyclonal antibody against IYSV. Comparative analyses of their sensitivity and specificity in detecting IYSV from infected tobacco leaves identified 7F4 as the best-performed MAb in MIA. We then optimized the working conditions of Luminex xMAP-based MIA in specific detection of IYSV from infected tobacco leaves by using appropriate blocking buffer and proper concentration of biotin-labeled antibodies as well as the suitable ratio between the antibodies and the streptavidin R-phycoerythrin (SA-RPE). Under the optimized conditions the Luminex xMAP-based MIA was able to specifically detect IYSV with much higher sensitivity than conventional enzyme-linked immunosorbent assay (ELISA). Importantly, the Luminex xMAP-based MIA is time-saving and the whole procedure could be completed within 2.5 h. CONCLUSIONS: We generated five specific MAbs against IYSV and developed the Luminex xMAP-based MIA method for specific detection of IYSV in plants. This assay provides a sensitive, high-specific, easy to perform and likely cost-effective approach for IYSV detection from infected plants, implicating potential broad usefulness of MIA in plant virus diagnosis.


Asunto(s)
Infecciones por Bunyaviridae/diagnóstico , Infecciones por Bunyaviridae/virología , Inmunoensayo , Mediciones Luminiscentes , Microesferas , Enfermedades de las Plantas/virología , Tospovirus/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Femenino , Inmunoensayo/métodos , Mediciones Luminiscentes/métodos , Ratones , Proteínas Recombinantes , Sensibilidad y Especificidad , Tospovirus/genética , Proteínas Virales/genética , Proteínas Virales/inmunología
17.
J Antibiot (Tokyo) ; 70(11): 1043-1046, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28928473

RESUMEN

Hymerhabdrin A (1), a diterpenoid possessing a novel 6/6/5 fused-ring skeleton, together with four known sterols were isolated from an intertidal marine sponge Hymerhabdia sp. Their structures were elucidated by extensive spectroscopic methods, and the relative and absolute configurations of 1 were determined by NOESY analysis and electronic circular dichrosim calculations, respectively. Hymerhabdrin A (1) exhibited significant antifouling activity against Balanus amphitrite larval with LC50 (lethal concentration 50) value of 3.6 µg ml-1.


Asunto(s)
Incrustaciones Biológicas/prevención & control , Diterpenos/farmacología , Poríferos/química , Animales , Dicroismo Circular/métodos , Diterpenos/química , Diterpenos/aislamiento & purificación , Dosificación Letal Mediana , Espectroscopía de Resonancia Magnética/métodos , Análisis Espectral/métodos , Esteroles/química , Esteroles/aislamiento & purificación , Thoracica/efectos de los fármacos
18.
J Virol Methods ; 243: 113-119, 2017 05.
Artículo en Inglés | MEDLINE | ID: mdl-28109844

RESUMEN

Cucumber green mottle mosaic virus (CGMMV)causes a severe mosaic symptom of watermelon and cucumber, and can be transmitted via infected cucumber seeds, leaves and soil. It remains a challenge to detect this virus to prevent its introduction and infection and spread in fields. For this purpose, a simple and sensitive label-free colorimetric detection method for CGMMV has been developed with unmodified gold nanoparticles (AuNPs) as colorimetric probes. The method is based on the finding that the presence of RT-PCR target products of CGMMV and species-specific probes results in color change of AuNPs from red to blue after NaCl induction. Normally, species-specific probes attach to the surface of AuNPs and thereby increasing their resistance to NaCl-induced aggregation. The concentration of sodium, probes in the reaction system and evaluation of specificity and sensitivity of a novel assay, visual detection of Cucumber green mottle mosaic virus using unmodified AuNPs has been carried out with simple preparation of samples in our study. Through this assay, as low as 30pg/µL of CGMMV RNA was thus detected visually, by the naked eye, without the need for any sophisticated, expensive instrumentation and biochemical reagents. The specificity was 100% and exhibited good reproducibility in our assays. The results note that this assay is highly species-specific, simple, low-cost, and visual for easy detection of CGMMV in plant tissues. Therefore, visual assay is a potentially useful tool for middle or small-scales corporations and entry-exit inspection and quarantine bureau to detect CGMMV in cucumber seeds or plant tissues.


Asunto(s)
Colorimetría/métodos , Oro , Nanopartículas , Enfermedades de las Plantas/virología , ARN Viral/análisis , Tobamovirus/aislamiento & purificación , Citrullus/virología , Cucumis sativus/virología , ARN Viral/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Cloruro de Sodio/metabolismo , Tobamovirus/genética
19.
World J Microbiol Biotechnol ; 32(9): 143, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27430510

RESUMEN

Biofilm formation can make significant effects on bacteria habits and biological functions. In this study, diketopiperazines (DKPs) produced by strain of Bacillus amyloliquefaciens Q-426 was found to inhibit biofilm formed in the gas-liquid interface. Four kinds of DKPs were extracted from B. amyloliquefaciens Q-426, and we found that 0.04 mg ml(-1) DKPs could obviously inhibit the biofilm formation of the strain. DKPs produced by B. amyloliquefaciens Q-426 made a reduction on extracellular polymeric substance (EPS) components, polysaccharides, proteins, DNAs, etc. Real-time PCR was performed to determine that whether DKPs could make an obvious effect on the expression level for genes related to biofilm formation in the strain. The relative expression level of genes tasA, epsH, epsG and remB which related to proteins, extracellular matrix, and polysaccharides, were downregulated with 0.04 mg ml(-1) DKPs, while the expression level of nuclease gene nuc was significantly upregulated. The quantitative results of the mRNA expression level for these genes concerted with the quantitative results on EPS levels. All of the experimental results ultimately indicated that DKPs could inhibit the biofilm formation of the strain B. amyloliquefaciens Q-426.


Asunto(s)
Bacillus amyloliquefaciens/efectos de los fármacos , Proteínas Bacterianas/genética , Dicetopiperazinas/farmacología , Bacillus amyloliquefaciens/genética , Bacillus amyloliquefaciens/fisiología , Biopelículas/efectos de los fármacos , Regulación hacia Abajo , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos
20.
Chem Commun (Camb) ; 52(56): 8707-10, 2016 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-27331773

RESUMEN

Based on an endonuclease-assisted, cross-triggered and cascaded recycling amplification strategy, the construction of a simple electrochemical sensing platform for the ultrasensitive detection of the mutant p53 gene in human serum is described. Using this new signal amplification approach, the sub-femtomolar level of the mutant p53 gene can be selectively detected.


Asunto(s)
Técnicas Electroquímicas , Mutación , Técnicas de Amplificación de Ácido Nucleico , Proteína p53 Supresora de Tumor/sangre , Proteína p53 Supresora de Tumor/genética , Humanos
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