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1.
Plants (Basel) ; 13(5)2024 Feb 22.
Artículo en Inglés | MEDLINE | ID: mdl-38475443

RESUMEN

Sucrose non-fermenting-1-related protein kinase-1 (SnRK1) is a highly conserved serine-threonine kinase complex regulating plants' energy metabolisms and resistance to various types of stresses. However, the downstream genes regulated by SnRK1 in these plant physiological processes still need to be explored. In this study, we found that the knockout of OsSnRK1a resulted in no obvious defects in rice growth but notably decreased the seed setting rate. The ossnrk1a mutants were more sensitive to blast fungus (Magnaporthe oryzae) infection and showed compromised immune responses. Transcriptome analyses revealed that SnRK1a was an important intermediate in the energy metabolism and response to biotic stress. Further investigation confirmed that the transcription levels of OsNADH-GOGAT2, which positively controls rice yield, and the defense-related gene pathogenesis-related protein 1b (OsPR1b) were remarkably decreased in the ossnrk1a mutant. Moreover, we found that OsSnRK1a directly interacted with the regulatory subunits OsSnRK1ß1 and OsSnRK1ß3, which responded specifically to blast fungus infection and starvation stresses, respectively. Taken together, our findings provide an insight into the mechanism of OsSnRK1a, which forms a complex with specific ß subunits, contributing to rice seed set and resistance by regulating the transcription of related genes.

2.
BMC Cancer ; 24(1): 268, 2024 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-38408959

RESUMEN

BACKGROUND: Gemcitabine is a cornerstone drug for the treatment of all stages of pancreatic cancer and can prolong the survival of patients with pancreatic cancer, but resistance to gemcitabine in pancreatic cancer patients hinders its efficacy. The overexpression of Early growth response 1(EGR1) in pancreatic ductal adenocarcinoma as a mechanism of gemcitabine chemoresistance in pancreatic cancer has not been explored. The major mechanisms of gemcitabine chemoresistance are related to drug uptake, metabolism, and action. One of the common causes of tumor multidrug resistance (MDR) to chemotherapy in cancer cells is that transporter proteins increase intracellular drug efflux and decrease drug concentrations by inducing anti-apoptotic mechanisms. It has been reported that gemcitabine binds to MDR1 with high affinity. The purpose of this research was to investigate the potential mechanisms by which EGR1 associates with MDR1 to regulate gemcitabine resistance in pancreatic cancer cells. METHODS: The following in vitro and in vivo techniques were used in this research to explore the potential mechanisms by which EGR1 binds to MDR1 to regulate gemcitabine resistance in pancreatic cancer cells. Cell culture; in vitro and in vivo study of EGR1 function by loss of function analysis. Binding of EGR1 to the MDR1 promoter was detected using the ChIP assay. qRT-PCR, Western blot assays to detect protein and mRNA expression; use of Annexin V apoptosis detection assay to test apoptosis; CCK8, Edu assay to test cell proliferation viability. The animal model of pancreatic cancer subcutaneous allograft was constructed and the tumours were stained with hematoxylin eosin and Ki-67 expression was detected using immunohistochemistry. FINDINGS: We revealed that EGR1 expression was increased in different pancreatic cancer cell lines compared to normal pancreatic ductal epithelial cells. Moreover, gemcitabine treatment induced upregulation of EGR1 expression in a dose- and time-dependent manner. EGR1 is significantly enriched in the MDR1 promoter sequence.Upon knockdown of EGR1, cell proliferation was impaired in CFPAC-1 and PANC-1 cell lines, apoptosis was enhanced and MDR1 expression was decreased, thereby partially reversing gemcitabine chemoresistance. In animal experiments, knockdown of EGR1 enhanced the inhibitory effect of gemcitabine on tumor growth compared with the sh-NC group. CONCLUSIONS: Our study suggests that EGR1 may be involved in the regulation of MDR1 to enhance gemcitabine resistance in pancreatic cancer cells. EGR1 could be a novel therapeutic target to overcome gemcitabine resistance in pancreatic cancer.


Asunto(s)
Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Animales , Humanos , Gemcitabina , Desoxicitidina/farmacología , Desoxicitidina/uso terapéutico , Resistencia a Antineoplásicos/genética , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Carcinoma Ductal Pancreático/tratamiento farmacológico , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patología , Línea Celular Tumoral , Apoptosis , Proteína 1 de la Respuesta de Crecimiento Precoz/genética
3.
Curr Pharm Des ; 30(3): 230-239, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38243946

RESUMEN

BACKGROUND: Artesunate (ART) has been recognized to induce ferroptosis in various tumor phenotypes, including neuroendocrine tumors. We aimed to investigate the effects of ART on insulinoma and the underlying mechanisms by focusing on the process of ferroptosis. METHODS: The CCK8 and colony formation assays were conducted to assess the effectiveness of ART. Lipid peroxidation, glutathione, and intracellular iron content were determined to validate the process of ferroptosis, while ferrostatin-1 (Fer-1) was employed as the inhibitor of ferroptosis. Subcutaneous tumor models were established and treated with ART. The ferroptosis-associated proteins were determined by western blot and immunohistochemistry assays. Pathological structures of the liver were examined by hematoxylin-eosin staining. RESULTS: ART suppressed the growth of insulinoma both in vitro and in vivo. Insulinoma cells treated by ART revealed signs of ferroptosis, including increased lipid peroxidation, diminished glutathione levels, and ascending intracellular iron. Notably, ART-treated insulinoma cells exhibited a decline in the expressions of catalytic component solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4). These alterations were negated by Fer-1. Moreover, no hepatotoxicity was observed upon the therapeutic dose of ART. CONCLUSION: Artesunate might regulate ferroptosis of insulinoma cells through the SLC7A11/GPX4 pathway.


Asunto(s)
Ciclohexilaminas , Ferroptosis , Insulinoma , Neoplasias Pancreáticas , Fenilendiaminas , Humanos , Artesunato , Glutatión , Hierro , Sistema de Transporte de Aminoácidos y+
4.
Xenobiotica ; 53(4): 231-240, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-37272731

RESUMEN

In vitro metabolism of bicyclol was studied using liver microsomes, hepatocytes and human recombinant cytochrome P450 enzymes. Liquid chromatography-benchtop orbitrap mass spectrometry technique was utilised to identify the metabolites.A total of 19 metabolites, including 5 new metabolites (M2, M3, M4, M5 and M16) were tentatively identified. Among these metabolites, M6&M8 (demethylenation), M9&M10 (demethylation) and M19 (glucuronidation) were the major metabolites.In glutathione (GSH)-supplemented liver microsomes, 5 new GSH conjugates were found and tentatively identified. The formation was assumed to be through demethylenation of methylenedioxyphenyl to form catechol derivatives, which further underwent oxidation to form ortho-quinone intermediates, reacting with GSH to form stable adducts.CYP3A4 and 2C19 were demonstrated to be the major enzymes responsible for the bioactivation of bicyclol.This study provided valuable information on the metabolic fate of bicyclol in liver microsomes and hepatocytes, and the bioactivation pathways were reported for the first time, which would be helpful for us to understand the potential drug-drug interactions and the possible side effect of this drug.


Asunto(s)
Sistema Enzimático del Citocromo P-450 , Microsomas Hepáticos , Humanos , Microsomas Hepáticos/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Compuestos de Bifenilo/metabolismo , Hepatocitos/metabolismo , Glutatión/metabolismo , Cromatografía Líquida de Alta Presión
5.
ACS Appl Mater Interfaces ; 15(19): 23489-23500, 2023 May 17.
Artículo en Inglés | MEDLINE | ID: mdl-37139799

RESUMEN

The real-time detection of nitric oxide (NO) in living cells is essential to reveal its physiological processes. However, the popular electrochemical detection strategy is limited to the utilization of noble metals. The development of new detection candidates without noble metal species still maintaining excellent catalytic performance has become a big challenge. Herein, we propose a spinel oxide doped with heteroatom-Cu-doped Co3O4 (Cu-Co3O4) for the sensitive and selective detection of NO release from the living cells. The material is strategically designed with Cu occupying the tetrahedral (Td) center of Co3O4 through the formation of a Cu-O bond. The introduced Cu regulates the local coordination environment and optimizes the electronic structure of Co3O4, hybridizing with the N 2p orbital to enhance charge transfer. The CuTd site can well inhibit the current response to nitrite (NO2-), resulting in a high improvement in the electrochemical oxidation of NO. The selectivity of Cu-Co3O4 can be markedly improved by the pore size of the molecular sieve and the negative charge on the surface. The rapid transmission of electrons is due to the fact that Cu-Co3O4 can be uniformly and densely in situ grown on Ti foil. The rationally designed Cu-Co3O4 sensor displays excellent catalytic activity toward NO oxidation with a low limit of detection of 2.0 nM (S/N = 3) and high sensitivity of 1.9 µA nM-1 cm-2 in cell culture medium. The Cu-Co3O4 sensor also shows good biocompatibility to monitor the real-time NO release from living cells (human umbilical vein endothelial cells: HUVECs; macrophage: RAW 264.7 cells). It was found that a remarkable response to NO was obtained in different living cells when stimulated by l-arginine (l-Arg). Moreover, the developed biosensor could be used for real-time monitoring of NO released from macrophages polarized to a M1/M2 phenotype. This cheap and convenient doping strategy shows universality and can be used for sensor design of other Cu-doped transition metal materials. The Cu-Co3O4 sensor presents an excellent example through the design of proper materials to implement unique sensing requirements and sheds light on the promising strategy for electrochemical sensor fabrication.


Asunto(s)
Óxido Nítrico , Óxidos , Humanos , Óxidos/química , Células Endoteliales de la Vena Umbilical Humana
6.
Plant Dis ; 2023 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-36973909

RESUMEN

Brown rot caused by Monilinia spp. (anamorph Monilia) is a common disease in stone fruits worldwide, but the species in different hosts or regions may vary. Monilia mumecola is a recently identified species, only reported in some regions. Although the pathogen has been found on plum in Yunnan, Hubei, and Zhejiang provinces, and Chongqing municipality in China (Yin et al. 2015), it has not been reported in southeast coast of China. In May 2022, brown rot with grey spores on fruit was observed in a plum orchard with 15% disease incidence in Sanming City, Fujian Province, located in southeast coast of China. Four single-spored isolates were obtained from germinating conidia on the water agar for further investigation. The colonies on potato dextrose agar (PDA) were initially white, gradually turned gray to brown, with lobbed margins and rare sporulation. Average mycelial growth rate ranged from 0.74 to 1.08 cm/day at 25 oC. Conidia were lemon-shaped or subglobose, hyaline, with an average size of 17.64 to 19.35×11.14 to 14.44 µm (n=30). Each isolate produced one to three or four germ tubes. Such characteristics are similar to M. mumecola (Yin et al. 2015). To confirm the identity of the isolates, genomic DNA was extracted and species-specific primers of Hu et al. (2011) were used to amplify the 712 bp sequence. In addition, the ITS region, partial genes of glyceraldehyde-3- phosphate dehydrogenase (GAPDH) and beta-tubulin (TUB2) were also amplified using primers sets ITS1/ITS4 (Glass and Donaldson 1995), Mon-G3pdhF/Mon-G3pdhR and Mon-TubF1/ Mon-TubR1, respectively (Hu et al. 2011). Sequences obtained for those three regions were 478, 762 and 1527 bp, respectively. Each region of all four isolates was identical, so one sequence for each region was submitted to GenBank with accession numbers OQ207672, OQ225251 and OQ225252, respectively, which had 100% identity with M. mumecola HQ908786 (ITS), HQ908784 (GAPDH), and HQ908775 (TUB2) using BLAST analysis in NCBI database, respectively. Pathogenicity was conducted with mycelium plugs from the edge of 7-day-old colony on three mature 'Angeleno' plums fruit (Prunus salicina) creating nine inoculations with three wounds per fruit, and each wound was 5 mm in diameter and 2 mm in depth. The same amount fruit and wounds inoculated with PDA plugs without fungi were used as a control. Brown rot symptoms were observed on all inoculated plums 4 days post-inoculation under room temperature with 100% humidity, whereas control plums remained symptomless. Fungal colonies re-isolated from the lesions showed the same morphological features as the original isolate , thus fulfilling Koch's postulates. To our knowledge, this is the first report of M. mumecola on plum in Fujian Province of China. The findings in this studies have important management implications for local plum growers because more than one Monilia species have been reported, where only M. fructicola was present in this region.

7.
Neuroendocrinology ; 113(6): 641-656, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36758529

RESUMEN

INTRODUCTION: 5-Iodotubercidin, a type of purine derivative, has attracted increasing attention in tumor chemotherapy because of its potential as an antitumor agent in recent years. In this study, we confirmed the effects on apoptosis in insulinoma cell lines induced by 5-iodotubercidin and tried to illuminate the underlying mechanisms. METHODS: We used 5-iodotubercidin in the treatment of insulinoma cells and the cell proliferation was examined using CCK-8 assay, colony-forming assays, and insulinoma animal models. Cell apoptosis was examined using TUNEL assays and Western blotting. Cellular DNA damage was shown by comet assay and immunofluorescence. The expression of apoptosis-regulating proteins and DNA damage biomarker was investigated by Western blotting. Subcutaneous inoculation of the insulinoma cells into nude mice was to measure blood glucose, insulin levels, and tumor growth. ATM siRNA and p53 siRNA were used as loss-of-function targets to evaluate 5-iodotubercidin treatment. RESULTS: 5-Iodotubercidin inhibited the proliferation of insulinoma cells and induced DNA damage and cell apoptosis. Moreover, 5-iodotubercidin induced ATM and p53 activated. In vivo, 5-iodotubercidin inhibited the growth of Ins-1 and Min-6 cells xenografts in nude mice. CONCLUSION: 5-Iodotubercidin induces DNA damage leading to insulinoma cells apoptosis by activating ATM/p53 pathway. Therefore, this is a potential strategy for treating insulinoma.


Asunto(s)
Insulinoma , Neoplasias Pancreáticas , Ratones , Animales , Humanos , Insulinoma/tratamiento farmacológico , Insulinoma/metabolismo , Insulinoma/patología , Ratones Desnudos , Proteína p53 Supresora de Tumor/genética , Apoptosis , Proliferación Celular , Neoplasias Pancreáticas/tratamiento farmacológico , ARN Interferente Pequeño/farmacología , Línea Celular Tumoral
8.
Pestic Biochem Physiol ; 188: 105262, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36464367

RESUMEN

Brown rot disease broke out in stone fruit orchards of Fujian, China in 2019, despite pre-harvest application of methyl benzimidazole carbamate (MBC). To determine the reason, a total of 44 Monilinia fructicola strains were collected from nectarine, plum and peach fruits in this study, among which 79.5% strains were resistant to thiophanate-methyl, indicated by discriminatory dose of 5 µg/mL. The resistance of these strains was confirmed by treating detached peach fruit with label rates of formulated thiophanate-methyl which only completely inhibit infection of the sensitive strains, but not the resistant strains. Further analysis of the mechanism of MBC resistance revealed that all resistant strains carry a H6Y mutation in ß-tubulin protein Tub2, which was only reported previously in the M. fructicola strains from California, USA, and do not display obvious fitness penalties, as no significant defects in mycelial growth rate, sporulation, conidia germination, aggressiveness on detached peach fruit and temperature sensitivity was detected. In addition, we found that diethofencarb, the agent for managing MBC-resistance strains, was unable to inhibit growth of the H6Y strains. Taken together, our study, for the first time, identified a mutation form of H6Y in the ß-tubulin protein of M. fructicola in China, rendering the strains wide resistance to thiophanate-methyl. This mechanism of M. fructicola gaining resistance to MBC fungicides needs to be fully considered, when designing management strategies to control brown rot disease in stone fruit orchards.


Asunto(s)
Prunus persica , Tiofanato , Tiofanato/farmacología , Tubulina (Proteína)/genética , Prevalencia , Prunus persica/genética , Mutación , China
10.
PLoS One ; 17(8): e0273605, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35994452

RESUMEN

[This corrects the article DOI: 10.1371/journal.pone.0231961.].

11.
Rapid Commun Mass Spectrom ; 36(19): e9362, 2022 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-35881078

RESUMEN

RATIONALE: Ligusticum chuanxiong Hort is a well-known herb medicine that has been widely prescribed to treat cardiovascular diseases in China for hundreds of years. Senkyunolide H (SNH) is one of the major bioactive ingredients extracted from L. chuanxiong, and it displayed neuroprotective effects. To fully understand its mechanism of action, the metabolism needs to be investigated. METHODS: In vitro studies were conducted by incubating SNH with rat and human hepatocytes, and the metabolites were identified and characterized using liquid chromatography in combination with hybrid quadrupole Orbitrap mass spectrometry (LC-Orbitrap-MS). The structures of the metabolites were proposed by accurate mass analysis of respective precursor ions, indicative product ions, and elemental compositions. RESULTS: Under the current conditions, a total of 10 metabolites were identified, and among these metabolites, M3 and M4 were the most abundant metabolites both in rat and human hepatocytes. Our results demonstrated that hydroxylation, hydration, glucuronidation, and GSH conjugation were the primary metabolic pathways of SNH. CONCLUSIONS: The present study provides new information on the metabolism of SNH, which would help prospects of the disposition of SNH.


Asunto(s)
Benzofuranos , Ligusticum , Animales , Benzofuranos/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Humanos , Ratas , Espectrometría de Masas en Tándem/métodos
12.
Comput Intell Neurosci ; 2022: 1761635, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35655490

RESUMEN

The main purpose of the object detection process is to determine the category of the scene object and use the display 3D and 3D frame size. At present, in the case of 3D object detection, we can extract more accurate features by learning a large number of data, and this deep learning network has good results, but there is a very big problem, including the error of input information, extraction error, and so on. Therefore, solving the above problems has become an important direction to promote the rapid development of 3D target detection technology. This paper mainly studies the deep learning wireless sensor technology and also studies the deep learning infrared and visible image fusion. At the same time, based on the introduction of wireless sensor technology and research status, this paper summarizes the existing algorithms. Texture image classification is a more important visual cue in life. Because it will be affected by light intensity, noise size, image scale, and so on. This makes the classification and feature extraction of image scale and texture image more difficult. To solve these problems has become a hot topic of computer vision research in recent years. The shape of the point cloud is completed by using the 3D target detection method to complete the algorithm research. The radar point cloud is extracted by the 3D target detection method, and the radar point group of the overall shape of the object is obtained. The principal component analysis algorithm is used to extract the principal features of the radar point cloud with the complete shape of the object, and the more accurate 3D target frame is obtained after feature adjustment.


Asunto(s)
Aprendizaje Profundo , Algoritmos , Radar , Tecnología , Tecnología Inalámbrica
13.
World J Microbiol Biotechnol ; 38(9): 147, 2022 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-35773599

RESUMEN

The presence of herbicides residues in soil represents a serious problem for agriculture. Quinclorac is a common herbicide applied in rice field, but its residue can cause abnormal growth in successive crop of tobacco in Southern China. Remediation by microorganisms is considered to be an environmentally friendly method to remove such pollutants injury. The aims of this study were to obtain quinclorac remediation isolates and to investigate the possible mechanism(s) of remediation. Six bacterial isolates were obtained from rhizosphere of rice-tobacco rotation fields, and were found to be capable of degrading quinclorac on a mineral salt medium (MSM), with degradation efficiency ranging from 2.1 to 23.7%. Among these isolates, J5 had the highest degradation efficiency, and was identified as Klebsiella variicola based on phylogenetic analyses and a metabolic profile generating by Biolog GEN III system. Bioremediation of quinclorac injury was confirmed using pot assays with tobacco, in which J5 reversed the detrimental effect of quinclorac on leaf area, leaf number, and plant height. The J5 isolate also seemed to promote plant growth, in terms of tobacco seedling growth and seed germination, which were 2.2 times and 1.6 times higher compared to untreated control, respectively. The mechanisms of plant growth promoting (PGP) traits were found to involve nitrogen-fixing, indole-3-acetic acid (IAA) production, and phosphate solubilization ability. In addition, proteomic analysis and relative quantitative PCR revealed an elevated level of 4-hydroxyphenylacetate 3-monooxygenase (HPMO) in quinclorac-treated J5, suggesting that this enzyme may play an important role in quinclorac remediation. This study showed that the J5 isolate could be exploited to not only assist in soil remediation due to quinclorac residue issues but also promote tobacco growth.


Asunto(s)
Herbicidas , Oryza , Bacterias/metabolismo , Biodegradación Ambiental , Herbicidas/metabolismo , Oryza/metabolismo , Filogenia , Proteómica , Quinolinas , Rizosfera , Suelo , Microbiología del Suelo , Nicotiana
14.
Spectrochim Acta A Mol Biomol Spectrosc ; 278: 121335, 2022 Oct 05.
Artículo en Inglés | MEDLINE | ID: mdl-35526438

RESUMEN

Phthalate esters (PAEs) are widely used as plasticizer components in production. Methyl hydrogen phthalate (MHP) is a metabolite of dimethyl phthalate (DMP, a kind of PAEs), and its toxic residues accumulate in the nature and can enter the human body. Here, the interaction between MHP and human serum albumin (HSA) was probed by using multi-spectral, computer simulations, and biochemical techniques. The results showed that MHP was spontaneously embedded in site I of HSA to form a complex by H-bonds and van der Waals forces (ΔH < 0, ΔS < 0). The binding constant (Ka) of the HSA-MHP system was 1.136 ± 0.026 × 104 M-1 (298 K). The combination of MHP produced conformational variations of HSA, as shown by the 3D fluorescence spectrum, CD spectra, and molecular dynamics simulation. Additionally, molecular docking indicated that MHP was surrounded by multiple residues, such as Lys199, Leu203, Phe206, and Trp214. Specifically, Lys199 and Trp214 exerted a crucial effect on the interaction of HSA and MHP. The residues with important energy contribution were mostly located in site I. The ASA values of the aromatic amino acids of HSA changed after combining with MHP. The Rg and SASA values of HSA increased after adding MHP, suggesting that the structure of HSA was less compact. Moreover, the esterase-like activity of HSA increased after adding MHP to HSA, indicating that MHP may disturb the normal physiological activities in the human body. This study was helpful to understand the biological function of MHP and provided some insights for its side effect in the human body.


Asunto(s)
Hidrógeno , Albúmina Sérica Humana , Sitios de Unión , Dicroismo Circular , Ésteres , Humanos , Simulación del Acoplamiento Molecular , Ácidos Ftálicos , Unión Proteica , Albúmina Sérica/química , Albúmina Sérica Humana/química , Espectrometría de Fluorescencia/métodos , Termodinámica
15.
Anal Chim Acta ; 1207: 339815, 2022 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-35491044

RESUMEN

Here, a colorimetric aptasensor was constructed for sensitively detecting quinclorac (QNC), a common herbicide. The aptasensor involved a novel amplification strategy and a classical strand displacement strategy. The amplification strategy, termed exonuclease III (Exo III)-assisted cyclic release of phosphorodiamidate morpholino oligomer (PMO) mimic enzyme strategy, was developed based on two new findings on PMO: 1) DNA hybridized with PMO could resist Exo III digestion; 2) a designed G-rich PMO (named P2) could bind to hemin to form a G-quadruplex PMOzyme with peroxidase-like activity. In this strategy, a designed DNA-PMO duplex (D1-P1) completely hybridized with DNA2 (D2) in the other designed DNA-PMO duplex (D2-P2) to trigger D2 degradation by Exo III and cyclic release of P2. After that, the hemin-binding P2 catalyzed colorless tetra-methyl benzidine (TMB) to blue TMB+. The cycle process was performed at high Exo III concentrations without strict control and with constant background signals. In that case, the developed strategy was sensitive, efficient, easy to operate, reliable, and ultralow background. Meanwhile, a QNC aptamer was used to develop the strand displacement strategy based on magnetic beads. The colorimetric aptasensor was sensitive and selective for QNC detection with a detection limit of 7.1 ng mL-1. It was successfully applied to detect QNC in soil and river water with good recovery rates (92-98%) and a relative standard deviation (n = 3) <5%. The success of this study could provide a general reference strategy for developing sensitive aptasensors and other nucleic acid-related sensors.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Aptámeros de Nucleótidos/metabolismo , Colorimetría , ADN , Exodesoxirribonucleasas , Hemina , Morfolinos , Quinolinas
16.
Plant Dis ; 2022 Mar 29.
Artículo en Inglés | MEDLINE | ID: mdl-35350891

RESUMEN

Potato black dot causing by Colletotrichum coccodes is a common disease in potato throughout the world, infecting underground stems, tubers, roots and foliage. Potato is becoming the third main food crop produced on ~16,000 ha annually in the Tibet Autonomous Region of China, situated on the world's highest plateau. However, the disease causing by C. coccodes has not been reported in this region. During the disease survey in the Bailang County of Tibet in August, 2020, some potato plants cv. "JiZhang 12" with chlorosis and wilting of foliage were observed. The incidence of affected plants was 20%. Necrotic lesions were also observed on the basal stems of the affected plants. Three affected plants were collected for pathogen isolation and three isolates were obtained for further investigation. The colonies on potato dextrose agar (PDA) were initially white, turning gradually black with age and producing abundant black sclerotia. Conidia were cylindrical, hyaline, aseptate, guttulate, with average size of 13.80 to 18.55×4.94 to 5.35 µm for the three isolates in which 30 conidia each were measured. Such characteristics are similar to C. coccodes (Lees and Hilton, 2003). Mycelial growth rate was 0.69 to 0.74 cm/day at 25 oC over the three isolates. The three isolates were confirmed to be C. coccodes by species-specific PCR using primer set of Cc1NF1/Cc2NR1 producing 350 bp amplicons in the internal transcribed spacer (ITS) region according to Cullen et al. (2002). The Cc1NF1/Cc2NR1 sequences were identical for three isolates, therefore one sequence from isolate BL_JZ_J1 was submitted to the GenBank with accession number OM368349. Additional genes were also sequencing including the actin (ACT), chitin synthase l (CHS1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and another larger ITS region were also amplified from genomic DNA using primers sets ACT512F/ACT783R, CHS-79F/CHS-354R (Carbone and Kohn 1999), GDF1/GDR1 (Templeton et al. 1992) and ITS1/ITS4 (Glass and Donaldson 1995), respectively. Sequences obtained for those four regions were 216 bp, 218bp, 235bp and 576 bp, respectively. Each region in the three isolates were also identical, therefore one sequence for each region was submitted to the GenBank with accession number of OM417059, OM417060, OM417061, and OM349570, respectively, which had 100% similarity with C. coccodes of MN336525 (ACT), KU821274 (CHS1), KU821397 (GAPDH) and KU821175 (ITS), respectively. Phylogenetic analyses based on the concatenated sequences of those four loci showed that the BL_JZ_J1 was close to C. coccodes, a reference isolate of CPOS1 with the accession numbers of GQ856787 (ACT), GQ856723 (CHS1), GQ856744 (GAPDH) and GQ485588 (ITS) (Yang et al. 2009). Pathogenicity was confirmed by inoculating a conidia suspension (100 µl of 105 conidia/ml) on three stems of 6-week-old potato plant cv. 'Favorita' with an artificial wound generated by sterile toothpick for each isolate. An equal volume of sterile water was injected on the wound of three stems as a noninoculated control. Brownish necrotic lesions were observed on all inoculated stems 3 days post-inoculation under natural conditions, whereas control stems remained symptomless. Reisolation of the fungus from all inoculated symptomatic plants confirmed Koch's Postulates. To our knowledge, this is the first report of C. coccodes in potato in Tibet Autonomous Region of China. The finding of black dot in this region has important management implications for the growers since the pathogen can survive for long periods in the field both on potato debris and in soil.

17.
J Photochem Photobiol B ; 228: 112394, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35086025

RESUMEN

Alcohol dehydrogenase (ADH) is a crucial enzyme in the alcohol metabolism pathway. Its activity is associated with the development of alcohol-relative diseases. Rutin is a kind of widely distributed dietary flavonoids, which have the ability to resist alcohol-induced liver injury. Here, the role of rutin on alcohol metabolism was investigated via the methods of biochemistry, spectroscopy and computer simulation. The experiment results demonstrated that rutin entered into the position of coenzyme (NAD) on ADH and formed a binary complex, which of process activated the catalyze activity of ADH in a concentration dependent manner. The combination of rutin on ADH induced microenvironmental variations as well as secondary structural change of ADH, where the level of α-helix reduced yet ß-sheet raised. The values of ∆H and ∆S suggested that H-bonds and van der Waals force occupied vital roles in the stabilization of ADH-rutin complex. Furthermore, molecular docking results further confirmed that the H-bonds between the hydroxyl groups on the benzene rings of rutin and surrounding amino acid were beneficial to maintain the stability of complex. Particularly, the van der Waals force and π-alkyl between rutin and Val residues may be the main reason for activation of ADH activity.


Asunto(s)
Alcohol Deshidrogenasa , Rutina , Alcohol Deshidrogenasa/química , Alcohol Deshidrogenasa/metabolismo , Simulación por Computador , Modelos Moleculares , Simulación del Acoplamiento Molecular , Análisis Espectral
18.
Anal Chem ; 94(2): 1076-1084, 2022 01 18.
Artículo en Inglés | MEDLINE | ID: mdl-34985279

RESUMEN

Closely related to multiple chronic inflammation, especially type-2 diabetes (T2D), methylglyoxal (MGO) may be a potential key to visualize disease progression and treatment effects. On the other hand, lack of convenient and fast analytical methods cannot afford accurate MGO quantitative evaluation. In this work, an activatable second near-infrared region (NIR-II) fluorescent probe TDTCD was synthesized and its reaction mechanism with MGO was discussed. The desired NIR-II product preferred response solvents with small polarity. A novel activatable nanoprobe, MG-SLNP, for MGO was then constructed based on rational packaging to provide a local nonpolar microenvironment. The hydrophobic core of nanoparticles not only successfully improved the stability and water solubility but also greatly promoted the response rate while reacting with MGO. The comparison between NIR-II fluorescence and the traditional high-performance liquid chromatography method for T2D blood samples was discussed. A high-resolution viewing window, quick response, and good biocompatibility led to a satisfactory signal-to-noise ratio of MG-SLNP for real-time MGO bio-detection and imaging in vivo.


Asunto(s)
Nanopartículas , Piruvaldehído , Colorantes Fluorescentes/química , Nanopartículas/química , Imagen Óptica/métodos
19.
Luminescence ; 37(1): 4-13, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34499419

RESUMEN

Resorcinol (1,3-dihydroxybenzene) is a common coupling agent in permanent hair dyes, and has arrested people's attention for its potential hazard to human health. However, the action mechanism of resorcinol and human DNA has not been elucidated. In this research, the binding properties between resorcinol and calf thymus DNA (ct-DNA) were studied for the first time through various spectral and molecular docking techniques. Spectral studies showed that the initial fluorescence quenching of resorcinol against DNA was a static one. The result of ΔH < 0 and ΔS > 0 was produced from thermodynamic experimental data, therefore it could be concluded that electrostatic force was the major driving force, while binding constant Kb was 1.56 × 104 M-1 at 298 K. The electrostatic binding network between resorcinol and ct-DNA was established explicitly through competitive substitution analysis and other spectral approaches. The results of FT-IR absorption spectra indicated that resorcinol had bound to the DNA phosphate skeleton. Molecular docking clearly revealed that binding occurred between hydroxyl groups of resorcinol and phosphorus oxygen bonds (P-O) of the DNA skeleton. These findings may deepen our understanding of the action mechanism between resorcinol and ct-DNA and provide some useful data on the effect of resorcinol on human diseases.


Asunto(s)
ADN , Resorcinoles , Dicroismo Circular , Humanos , Simulación del Acoplamiento Molecular , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier
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