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Flowering time, maturity, and plant height are crucial agronomic traits controlled by photoperiod that affect soybean (Glycine max [L.] Merr.) yield and regional adaptability. It is important to cultivate soybean cultivars of earlier maturity that adapt to high latitudes. GAMYB-binding protein 1 (GmGBP1), a member of the SNW/SKIP family of transcriptional coregulators in soybean, is induced by short days and interacts with transcription factor GAMYB (GmGAMYB) during photoperiod control of flowering time and maturity. In the present study, GmGBP1:GmGBP1 soybean showed the phenotypes of earlier maturity and higher plant height. Chromatin immunoprecipitation sequencing (ChIP-seq) assays of GmGBP1-binding sites and RNA sequencing (RNA-seq) of differentially expressed transcripts in GmGBP1:GmGBP1 further identified potential targets of GmGBP1, including small auxin-up RNA (GmSAUR). GmSAUR:GmSAUR soybean also showed earlier maturity and higher plant height. GmGBP1 interacted with GmGAMYB, bound to the promoter of GmSAUR and promoted the expression of FLOWER LOCUS T homologs 2a (GmFT2a) and FLOWERING LOCUS D LIKE 19 (GmFDL19). Flowering repressors such as GmFT4 were negatively regulated, resulting in earlier flowering and maturity. Furthermore, the interaction of GmGBP1 with GmGAMYB increased the gibberellin (GA) signal to promote height and hypocotyl elongation by activating GmSAUR and GmSAUR bound to the promoter of the GA-positive activating regulator gibberellic acid-stimulated Arabidopsis 32 (GmGASA32). These results suggested a photoperiod regulatory pathway in which the interaction of GmGBP1 with GmGAMYB directly activated GmSAUR to promote earlier maturity and plant height in soybean.
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Glycine max , Proteínas de Plantas , Factores de Transcripción , Hipocótilo/metabolismo , Fotoperiodo , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , ARN de Planta/genética , Transducción de Señal , Glycine max/metabolismo , Factores de Transcripción/metabolismo , Transcripción GenéticaRESUMEN
Metabolic reprogramming mediates antibiotic efficacy. However, metabolic adaptation of microbes evolving from antibiotic sensitivity to resistance remains undefined. Therefore, untargeted metabolomics was conducted to unveil relevant metabolic reprogramming and potential intervention targets involved in gentamicin resistance. In total, 61 metabolites and 52 metabolic pathways were significantly altered in gentamicin-resistant E. coli. Notably, the metabolic reprogramming was characterized by decreases in most metabolites involved in carbohydrate and amino acid metabolism, and accumulation of building blocks for nucleotide synthesis in gentamicin-resistant E. coli. Meanwhile, fatty acid metabolism and glycerolipid metabolism were also significantly altered in gentamicin-resistant E. coli. Additionally, glycerol, glycerol-3-phosphate, palmitoleate, and oleate were separately defined as the potential biomarkers for identifying gentamicin resistance in E. coli. Moreover, palmitoleate and oleate could attenuate or even abolished killing effects of gentamicin on E. coli, and separately increased the minimum inhibitory concentration of gentamicin against E. coli by 2 and 4 times. Furthermore, palmitoleate and oleate separately decreased intracellular gentamicin contents, and abolished gentamicin-induced accumulation of reactive oxygen species, indicating involvement of gentamicin metabolism and redox homeostasis in palmitoleate/oleate-promoted gentamicin resistance in E. coli. This study identifies the metabolic reprogramming, potential biomarkers and intervention targets related to gentamicin resistance in bacteria.
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Antibacterianos , Farmacorresistencia Bacteriana , Escherichia coli , Ácidos Grasos Monoinsaturados , Gentamicinas , Ácido Oléico , Gentamicinas/farmacología , Gentamicinas/metabolismo , Escherichia coli/metabolismo , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Ácido Oléico/metabolismo , Ácido Oléico/farmacología , Farmacorresistencia Bacteriana/efectos de los fármacos , Antibacterianos/farmacología , Ácidos Grasos Monoinsaturados/metabolismo , Ácidos Grasos Monoinsaturados/farmacología , Pruebas de Sensibilidad Microbiana , Metabolómica/métodos , Redes y Vías Metabólicas/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Flowering time and maturity are crucial agronomic traits that affect the regional adaptability of soybean plants. The development of soybean cultivars with early maturity adapted to longer days and colder climates of high latitudes is very important for ensuring normal ripening before frost begins. FUL belongs to the MADS-box transcription factor family and has several duplicated members in soybeans. In this study, we observed that overexpression of GmFULc in the Dongnong 50 cultivar promoted soybean maturity, while GmFULc knockout mutants exhibited late maturity. Chromatin immunoprecipitation sequencing (ChIP-seq) and RNA sequencing (RNA-seq) revealed that GmFULc could bind to the CArG, bHLH and homeobox motifs. Further investigation revealed that GmFULc could directly bind to the CArG motif in the promoters of the GmZTL3 and GmZTL4 genes. Overexpression of GmZTL4 promoted soybean maturity, whereas the ztl4 mutants exhibited delayed maturity. Moreover, we found that the cis element box 4 motif of the GmZTL4 promoter, a motif of light response elements, played an important role in controlling the growth period. Deletion of this motif shortened the growth period by increasing the expression levels of GmZTL4. Functional investigations revealed that short-day treatment promoted the binding of GmFULc to the promoter of GmZTL4 and inhibited the expression of E1 and E1Lb, ultimately resulting in the promotion of flowering and early maturation. Taken together, these findings suggest a novel photoperiod regulatory pathway in which GmFULc directly activates GmZTL4 to promote earlier maturity in soybean.
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Regulación de la Expresión Génica de las Plantas , Glycine max , Proteínas de Dominio MADS , Proteínas de Plantas , Glycine max/genética , Glycine max/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas de Dominio MADS/genética , Proteínas de Dominio MADS/metabolismo , Regiones Promotoras Genéticas/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Transcripción Genética , Plantas Modificadas Genéticamente/genéticaRESUMEN
Green strategy for the preparation of copper oxide nanoparticles (CuO NPs) using table olive has been researched in the present work. Some characterization assays viz., transmission electron microscopy (TEM), X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy-dispersive X-ray (EDX) was used for evaluation of the crystal structure, size and morphology of the manufactured NPs. As a catalyst, the prepared material demonstrated remarkable catalytic capability (>99% in 4 min) for the reduction of rhodamine B using sodium borohydride. In addition, the treated cells with the CuO NPs were examined by regarding the cytotoxicity properties on normal (HUVEC) cell line. The results showed that the prepared CuO NPs did not have any cytotoxicity effects on HUVEC (up to 500 µg/mL). Furthermore, in vivo experiments on burn wounds in rats show that the synthesized CuO NPs ointment significantly diminished (p ≤ 0.01) the wound area. On the other hand, the wound contracture factor was increased in comparison with the control groups. Collectively, the CuO NPs prepared by biological method have potential applications in organic pollutants reduction and wound care applications. In this viewpoint, CuO NPs may be considered as an effective for treatment of different wounds including burn wounds or injuries from surgeries such as plastic surgery.
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Graphene is an emerging nanomaterial increasingly being used in electrochemical biosensing applications owing to its high surface area, excellent conductivity, ease of functionalization, and superior electrocatalytic properties compared to other carbon-based electrodes and nanomaterials, enabling faster electron transfer kinetics and higher sensitivity. Graphene electrochemical biosensors may have the potential to enable the rapid, sensitive, and low-cost detection of cancer biomarkers. This paper reviews early-stage research and proof-of-concept studies on the development of graphene electrochemical biosensors for potential future cancer diagnostic applications. Various graphene synthesis methods are outlined along with common functionalization approaches using polymers, biomolecules, nanomaterials, and synthetic chemistry to facilitate the immobilization of recognition elements and improve performance. Major sensor configurations including graphene field-effect transistors, graphene modified electrodes and nanocomposites, and 3D graphene networks are highlighted along with their principles of operation, advantages, and biosensing capabilities. Strategies for the immobilization of biorecognition elements like antibodies, aptamers, peptides, and DNA/RNA probes onto graphene platforms to impart target specificity are summarized. The use of nanomaterial labels, hybrid nanocomposites with graphene, and chemical modification for signal enhancement are also discussed. Examples are provided to illustrate applications for the sensitive electrochemical detection of a broad range of cancer biomarkers including proteins, circulating tumor cells, DNA mutations, non-coding RNAs like miRNA, metabolites, and glycoproteins. Current challenges and future opportunities are elucidated to guide ongoing efforts towards transitioning graphene biosensors from promising research lab tools into mainstream clinical practice. Continued research addressing issues with reproducibility, stability, selectivity, integration, clinical validation, and regulatory approval could enable wider adoption. Overall, graphene electrochemical biosensors present powerful and versatile platforms for cancer diagnosis at the point of care.
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Grafito , Neoplasias , Reproducibilidad de los Resultados , Carbono , Anticuerpos , Biomarcadores de Tumor , Neoplasias/diagnósticoRESUMEN
Objective: This study aims to assess the effectiveness of surveillance inspections conducted by the provincial health committee in Quanzhou city during a COVID-19 outbreak in reducing false-positive results in SARS-CoV-2 RT-PCR assays. Method: The team conducted on-site inspections of laboratories that participated in mass screening, recording any violations of rules. Results: The positive cases in five rounds of mass screening were 23, 173, and 4 in Licheng District, Fengze District, and Luojang District, respectively. The false-positive rates in the five rounds of mass screening were 0.0099%, 0.0063%, 0.0018%, 0.0006%, and 0%, respectively. The study also recorded that the number of violations in the seven selected laboratories was 36, 68, 69, 42, 60, 54 and 47. The corresponding false-positive rates were 0.0012%, 0.0060%, 0.0082%, 0.0032%, 0.0060%, 0.0027%, and 0.0021%, respectively. The study found a positive correlation between false-positive rates and the number of violations (r = 0.905, P=0.005), and an inverse correlation between false-positive rates and the frequency of surveillance inspections (r = -0.950, P < 0.001). Conclusion: Daily surveillance inspection in laboratories can remind laboratories to strictly comply with standard procedures, focus on laboratory quality control, and reduce the occurrence of false-positive cases in SARS-CoV-2 nucleic acid tests to some extent. This study recommends that government decision-making departments establish policies and arrange experts to conduct daily surveillance inspections to improve laboratory quality control.
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BACKGROUND: Fentanyl is an analgesic used against pancreatitis-related pain, while whether it ameliorates severe acute pancreatitis (SAP) has yet to be checked. This study aims to determine fentanyl-delivered effect on SAP and the mechanism underlying this effect. METHODS: Rat SAP models were established, following fentanyl treatment. The serum activity of amylase (AMY), lipase (LIP), and diamine oxidase (DAO) was detected by enzyme-linked immunosorbent assay (ELISA). Histological examination was performed in the pancreatic and intestinal tissues with hematoxylin-eosin staining. After transfection with matrix metalloproteinase (MMP) 9 overexpression plasmids, Caco-2 monolayers were treated with fentanyl and subsequently exposed to lipopolysaccharide (LPS). The transepithelial electrical resistance (TEER) value was determined in rat intestinal mucosa through an Ussing chamber assisted by Analyze & Acquire, and in Caco-2 cell monolayers through a voltohmmeter. Intestinal mucosa and paracellular permeabilities were determined by fluorescein isothiocyanate (FITC)-labeled dextran assay. The expressions of ZO-1, Occludin, MMP9, Fas and Fas ligand (FasL) in rat intestinal mucosa and/or Caco-2 monolayers were analyzed by qRT-PCR or/and western blot. RESULTS: Fentanyl alleviated SAP-related histological alterations in the pancreas and intestines, reduced the elevated levels of SAP-related AMY, LIP, and DAO, but promoted the levels of ZO-1 and Occludin. In SAP rats and Caco-2 monolayers, SAP-related or LPS-induced TEER value decreases, permeability increases, and increases in the expressions of MMP9, Fas, and FasL were reversed partly by fentanyl. Notably, MMP9 overexpression could reverse the above fentanyl-delivered in vitro effects. CONCLUSIONS: Fentanyl alleviates intestinal mucosal barrier damage in rats with SAP by inhibiting the MMP9/FasL/Fas pathway.
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Amina Oxidasa (conteniendo Cobre) , Pancreatitis , Enfermedad Aguda , Amina Oxidasa (conteniendo Cobre)/metabolismo , Amina Oxidasa (conteniendo Cobre)/farmacología , Amilasas/metabolismo , Animales , Células CACO-2 , Dextranos/metabolismo , Eosina Amarillenta-(YS)/metabolismo , Proteína Ligando Fas/metabolismo , Fentanilo/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Hematoxilina/metabolismo , Hematoxilina/farmacología , Humanos , Mucosa Intestinal , Lipasa/metabolismo , Lipopolisacáridos/farmacología , Metaloproteinasa 9 de la Matriz , Ocludina/metabolismo , Ocludina/farmacología , Pancreatitis/inducido químicamente , Pancreatitis/tratamiento farmacológico , Pancreatitis/metabolismo , RatasRESUMEN
The imbalance of oxidation and antioxidant systems in the biological system can lead to oxidative stress, which is closely related to the pathogenesis of many diseases. Substances with antioxidant capacity can effectively resist the harmful damage of oxidative stress. How to measure the antioxidant capacity of antioxidants has essential application value in medicine and food. Techniques such as DPPH radical scavenging have been developed to measure antioxidant capacity. However, these traditional analytical techniques take time and require large instruments. It is a more convenient method to evaluate the antioxidant capacity of antioxidants based on their electrochemical oxidation and reduction behaviors. This review summarizes the evaluation of antioxidants using electrochemical sensors by bibliometrics. The development of this topic was described, and the research priorities at different stages were discussed. The topic was investigated in 1999 and became popular after 2010 and has remained popular ever since. A total of 758 papers were published during this period. In the early stages, electrochemical techniques were used only as quantitative techniques and other analytical techniques. Subsequently, cyclic voltammetry was used to directly study the electrochemical behavior of different antioxidants and evaluate antioxidant capacity. With methodological innovations and assistance from materials science, advanced electrochemical sensors have been fabricated to serve this purpose. In this review, we also cluster the keywords to analyze different investigation directions under the topic. Through co-citation of papers, important papers were analyzed as were how they have influenced the topic. In addition, the author's country distribution and category distribution were also interpreted in detail. In the end, we also proposed perspectives for the future development of this topic.
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Antioxidantes , Estrés Oxidativo , Antioxidantes/química , Bibliometría , Técnicas Electroquímicas/métodos , Oxidación-ReducciónRESUMEN
Electrochemical sensors have shown potential in recent years for plant species identification and phylogenetic studies. These works have been used to investigate the affinities of different species in many genera. However, the ability of electrochemical sensors to study relationships between different genera within a family has not been investigated. In this work, we selected 31 species in the Labiatae and 5 exotaxa as subjects to investigate the feasibility of electrochemical sensors at the genus level. The results show that electrochemical sensors are still very effective for the identification of these plants. Different pattern recognition techniques can make the identification more efficient. Also, the fingerprint profiles collected by the sensors can be used for phylogenetic studies of Labiatae. The phylogram divides all the species into five clusters, where the exotaxa are in one cluster. Species in the Labiatae are mainly distributed in four other clusters. Importantly, the different genera of species all showed close affinities, representing that electrochemical fingerprinting can well distinguish the affinities between the different genera. The results of this work demonstrate the great potential of electrochemical sensors in the study of plant phylogeny. Its application is not limited to the study at the species level, but can be extended to the genus level.
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Lamiaceae , Humanos , Filogenia , PlantasRESUMEN
Epstein-Barr virus (EBV) BamHI A rightward transcripts (BART) encoded microRNAs (EBV-miR-BARTs) are abnormally highly expressed in nasopharyngeal carcinoma (NPC). This study aims to investigate the diagnostic and prognostic performance of miR-BART7-3p and miR-BART13-3p. Plasma levels of EBV DNA, miR-BART7-3p, and miR-BART13-3p were examined by quantitative PCR in 483 treatment-naïve NPC patients and 243 controls without NPC. The prognostic performance was examined by comparing plasma levels with rates of distant metastasis during follow-up. The area under the receiver operating characteristic curve for diagnosing NPC was 0.926 for EBV DNA, 0.964 for plasma miR-BART7-3p, 0.973 for miR-BART13-3p, and 0.997 for all three indices. Among 465 NPC patients without distant metastasis, the above-median miR-BART7-3p and EBV DNA were independent risk for shorter distant metastasis-free survival (DMFS) (hazard ratio [HR] = 2.94, 95% confidence interval [CI], 1.44-5.97, P = .003; HR = 2.27, 95% CI, 1.26-4.10, P = .006) in multivariate Cox regression. Epstein-Barr virus DNA, miR-BART7-3p, and miR-BART13-3p after radiotherapy were detectable in 28.6%, 17.6%, and 54.7% of patients, respectively. In multivariate Cox regression, detectable miR-BART7-3p and EBV DNA were independent risks for shorter DMFS (HR = 4.13, 95% CI, 1.89-9.01, P < .001; HR = 2.14, 95% CI, 1.04-4.42, P = .039). The 4-year DMFS rate was 92.0% in subjects (n = 156) with neither detectable miR-BART7-3p nor EBV DNA, 80.0% in subjects (n = 65) with either detectable miR-BART7-3p or EBV DNA, and 52.9% in subjects (n = 24) with both detectable miR-BART7-3p and EBV DNA after radiotherapy (P < .001). Circulating levels of miR-BART7-3p and miR-BART13-3p show excellent diagnostic performance for NPC. The combination of plasma levels of miR-BART7-3p and EBV DNA at diagnosis and after radiotherapy could help stratify patients by risk of poor DMFS.
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MicroARN Circulante/sangre , Herpesvirus Humano 4/genética , Carcinoma Nasofaríngeo/diagnóstico , Neoplasias Nasofaríngeas/diagnóstico , ARN Viral/sangre , Adulto , Anciano , Biomarcadores de Tumor/sangre , ADN Viral/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Carcinoma Nasofaríngeo/sangre , Carcinoma Nasofaríngeo/patología , Carcinoma Nasofaríngeo/terapia , Neoplasias Nasofaríngeas/sangre , Neoplasias Nasofaríngeas/patología , Neoplasias Nasofaríngeas/terapia , Pronóstico , Análisis de Supervivencia , Adulto JovenRESUMEN
Soybean is the world's most important leguminous crop producing high-quality protein and oil. Elevating oil accumulation in soybean seed is always many researchers' goal. WRINKLED1 (WRI1) encodes a transcription factor of the APETALA2/ethylene responsive element-binding protein (AP2/EREBP) family that plays important roles during plant seed oil accumulation. In this study, we isolated and characterized three distinct orthologues of WRI1 in soybean (Glycine max) that display different organ-specific expression patterns, among which GmWRI1a was highly expressed in maturing soybean seed. Electrophoretic mobility shift assays and yeast one-hybrid experiments demonstrated that the GmWRI1a protein was capable of binding to AW-box, a conserved sequence in the proximal upstream regions of many genes involved in various steps of oil biosynthesis. Transgenic soybean seeds overexpressing GmWRI1a under the control of the seed-specific napin promoter showed the increased total oil and fatty acid content and the changed fatty acid composition. Furthermore, basing on the activated expressions in transgenic soybean seeds and existence of AW-box element in the promoter regions, direct downstream genes of GmWRI1a were identified, and their products were responsible for fatty acid production, elongation, desaturation and export from plastid. We conclude that GmWRI1a transcription factor can positively regulate oil accumulation in soybean seed by a complex gene expression network related to fatty acid biosynthesis.
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Glycine max/metabolismo , Aceites de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Semillas/metabolismo , Proteínas de Soja/metabolismo , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Ácidos Grasos/biosíntesis , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/clasificación , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Regiones Promotoras Genéticas/genética , Unión Proteica , Semillas/genética , Homología de Secuencia de Aminoácido , Proteínas de Soja/clasificación , Proteínas de Soja/genética , Glycine max/genética , Factores de Transcripción/clasificación , Factores de Transcripción/genéticaRESUMEN
BACKGROUND: Epstein-Barr virus capsid antigen immunoglobulin A (EBV VCA-IgA) exerts an important role in the diagnosis of nasopharyngeal carcinoma (NPC). This meta-analysis aimed to evaluate the pooled diagnostic performance of VCA-IgA for NPC. METHODS: Literature fulfilling the criteria was searched in PubMed and Embase databases. The quality of the studies was assessed in terms of the Quality Assessment of Diagnosis Accuracy Studies (QUADAS) criteria. The pooled diagnostic parameters were generated using a bivariate meta-analysis model. Statistical analysis was performed based on the platforms of Meta-Disc 1.4 and Stata 12.0 software. The trim and fill adjustment method was applied to further assess the possible effects of publication bias. RESULT: Twenty one studies comprising 2986 NPC patients and 3507 controls were included in this meta-analysis. The overall pooled sensitivity and specificity of serum VCA-IgA for NPC were 0.83 (95%CI: 0.82 - 0.84) and 0.88 (95% CI: 0.87 - 0.89), respectively, accompanied by a pooled diagnostic odds ratio (DOR) of 49.87 and area under curve (AUC) of 0.9390. Moreover, our stratified analyses suggested that combinations of multiple EBV antigens (sensitivity, specificity, DOR, and AUC of 0.93, 0.95, 331.8, and 0.9850, respectively) yielded higher accuracy than single VCA-IgA test (sensitivity, specificity, DOR and AUC of 0.83, 0.88, 49.87, and 0.9393, respectively). Additionally, the immunoenzyme assay (IEA) seemed to be a better alternative for the analysis of serum VCA-IgA level, with a sensitivity of 0.92, specificity of 0.94, and AUC of 0.9644. CONCLUSIONS: Serum VCA-IgA hallmarks promising accuracy in the management of NPC and that parallel tests of multiple EBV antigens may be more suitable for NPC serodiagnosis than single VCA-IgA assay. .151122)
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Anticuerpos Antivirales/sangre , Antígenos Virales/inmunología , Biomarcadores de Tumor/sangre , Proteínas de la Cápside/inmunología , Infecciones por Virus de Epstein-Barr/diagnóstico , Herpesvirus Humano 4/inmunología , Técnicas para Inmunoenzimas , Inmunoglobulina A/sangre , Neoplasias Nasofaríngeas/diagnóstico , Pruebas Serológicas , Área Bajo la Curva , Carcinoma , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Infecciones por Virus de Epstein-Barr/sangre , Infecciones por Virus de Epstein-Barr/inmunología , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/sangre , Neoplasias Nasofaríngeas/inmunología , Oportunidad Relativa , Valor Predictivo de las Pruebas , Curva ROCRESUMEN
The genes of sulfur-containing amino acid synthetases in soybean are essential for the synthesis of sulfur-containing amino acids. Gene mining of these enzymes is the basis for the molecular assistant breeding of high sulfur-containing amino acids in soybean. In this study, using software BioMercator2.1, 113 genes of sulfur-containing amino acid enzymes and 33 QTLs controlling the sulfur-containing amino acids content were mapped onto Consensus Map 4.0, which was integrated by genetic and physical maps of soybean. Sixteen candidate genes associated to the synthesis of sulfur-containing amino acids were screened based on the synteny between gene loci and QTLs, and the effect values of QTLs. Through a bioinformatic analysis of the copy number, SNP information, and expression profile of candidate genes, 12 related enzyme genes were identified and mapped on 8 linkage groups, such as D1a, M, A2, K, and G. The genes corresponding to QTL regions can explain 6%?38.5% genetic variation of sulfur-containing amino acids, and among them, the indirect effect values of 9 genes were more than 10%. These 12 genes were involved in sulfur-containing amino acid metabolism and were highly expressed in the cotyledons and flowers, showing an abundance of SNPs. These genes can be used as candidate genes for the development of functional markers, and it will lay a foundation for molecular design breeding in soybean.
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Aminoácidos/metabolismo , Glycine max/enzimología , Proteínas de Plantas/genética , Azufre/metabolismo , Transferasas/genética , Mapeo Cromosómico , Proteínas de Plantas/metabolismo , Polimorfismo de Nucleótido Simple , Sitios de Carácter Cuantitativo , Glycine max/genética , Glycine max/metabolismo , Transferasas/metabolismoRESUMEN
To establish a rapid detection method for norovirus GII.2 genotype, this study employed reverse transcription recombinase polymerase amplification (RT-RPA) combined with CRISPR/Cas12a and lateral flow strip (RT-RPA-Cas12a-LFS). Here, the genome of norovirus GII.2 genotype was compared to identify highly conserved sequences, facilitating the design of RT-RPA primers and crRNA specific to the conserved regions of norovirus GII.2. Subsequently, the reaction parameters of RT-RPA were optimized and evaluated using agar-gel electrophoresis and LFS. The results indicate that the conserved sequences of norovirus GII.2 were successfully amplified through RT-RPA at 37°C for 25 minutes. Additionally, CRISPR/Cas12a-mediated cleavage detection was achieved through LFS at 37°C within 10 minutes using the amplification products as templates. Including the isothermal amplification reaction time, the total time is 35 minutes. The established RT-RPA-Cas12a-LFS method demonstrated specific detection of norovirus GII.2, yielding negative results for other viral genomes, and exhibited an excellent detection limit of 10 copies/µl. The RT-RPA-Cas12a-LFS method was further compared with qRT-PCR by analyzing 60 food-contaminated samples. The positive conformity rate was 100%, the negative conformity rate was 95.45%, and the overall conformity rate reached 98.33%. This detection method for norovirus GII.2 genotype is cost-effective, highly sensitive, specific, and easy to operate, offering a promising technical solution for field-based detection of the norovirus GII.2 genotype.
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Genotipo , Norovirus , Norovirus/genética , Norovirus/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , Sistemas CRISPR-Cas , Humanos , ARN Viral/genética , Infecciones por Caliciviridae/virología , Infecciones por Caliciviridae/diagnóstico , Sensibilidad y EspecificidadRESUMEN
Soybean [Glycine max (L.) Merr.], an essential staple food and oil crop worldwide, boasts abundant vegetable proteins and fats beneficial for both human and animal consumption. However, the soybean pod borer (Leguminivora glycinivorella) (SPB) stands as the most destructive soybean insect pest in northeast China and other northeastern Asian regions, leading to significant annual losses in soybean yield and economic burden. Therefore, this study aims to investigate the introduction of a previously tested codon-optimized cry1c gene, cry1c*, into the soybean genome and assess its effect on the SPB infestation by generating and characterizing stable transgenic soybeans overexpressing cry1c*. The transgenic soybean lines that constitutively overexpressed cry1c* exhibited a significant reduction in the percentage of damaged seeds, reaching as low as 5% in plants under field conditions. Additionally, feeding transgenic leaves to the larvae of S. exigua, S. litura, and M. separta resulted in inhibited larval growth, decreased larval body weight, and lower survival rates compared to larvae fed on wild-type leaves. These findings showed that the transgenic lines maintained their resistance to SPB and other lepidopteran pests, especially the transgenic line KC1. Southern blotting and genome-wide resequencing analysis revealed that T-DNA integration occurred as a single copy between loci 50,868,122 and 50,868,123 of chromosome 10 in the transgenic line KC1. Therefore, the transgenic line KC1, overexpressing high levels of cry1c* in leaves and seeds, holds strong potential for commercial use in the integrated management of SPB and other lepidopteran pests.
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Soybean quality and production are determined by seed viability. A seed's capacity to sustain germination via dry storage is known as its seed life. Thus, one of the main objectives for breeders is to preserve genetic variety and gather germplasm resources. However, seed quality and germplasm preservation have become significant obstacles. In this study, four artificially simulated aging treatment groups were set for 0, 24, 72, and 120 hours. Following an aging stress treatment, the transcriptome and metabolome data were compared in two soybean lines with notable differences in seed vigor-R31 (aging sensitive) and R80 (aging tolerant). The results showed that 83 (38 upregulated and 45 downregulated), 30 (19 upregulated and 11 downregulated), 90 (52 upregulated and 38 downregulated), and 54 (25 upregulated and 29 downregulated) DEGs were differentially expressed, respectively. A total of 62 (29 upregulated and 33 downregulated), 94 (49 upregulated and 45 downregulated), 91 (53 upregulated and 38 downregulated), and 135 (111 upregulated and 24 downregulated) differential metabolites accumulated. Combining the results of transcriptome and metabolome investigations demonstrated that the difference between R31 and R80 responses to aging stress was caused by genes related to phenylpropanoid metabolism pathway, which is linked to the seed metabolite caffeic acid. According to this study's preliminary findings, the aging-resistant line accumulated more caffeic acid than the aging-sensitive line, which improved its capacity to block lipoxygenase (LOX) activity. An enzyme activity inhibition test was used to demonstrate the effect of caffeic acid. After soaking seeds in 1 mM caffeic acid (a LOX inhibitor) for 6 hours and artificially aging them for 24 hours, the germination rates of the R31 and R80 seeds were enhanced. In conclusion, caffeic acid has been shown to partially mitigate the negative effects of soybean seed aging stress and to improve seed vitality. This finding should serve as a theoretical foundation for future research on the aging mechanism of soybean seeds.
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A novel antiviral protein, designated as Stellarmedin A, was purified from Stellaria media (L.) Vill. (Caryophyllaceae) by using ammonium sulfate precipitation, cation-exchange chromatography system. Gel electrophoresis analysis showed that Stellarmedin A is a highly basic glycoprotein with a molecular weight of 35.1 kDa and an isoelectric point of â¼8.7. The N-terminal 14-amino acid sequence, MGNTGVLTGERNDR, is similar to those of other plant peroxidases. This protein inhibited herpes simplex virus type 2 (HSV-2) replication in vitro with an IC50 of 13.18 µg/ml and a therapeutic index exceeding 75.9. It was demonstrated that Stellarmedin A affects the initial stage of HSV-2 infection and is able to inhibit the proliferation of promyelocytic leukemia HL-60 and colon carcinoma LoVo cells with an IC50 of 9.09 and 12.32 µM, respectively. Moreover, Stellarmedin A has a peroxidase activity of 36.6 µmol/min/mg protein, when guaiacol was used as substrate. To our knowledge, this is the first report about an anti-HSV-2 protein with antiproliferative and peroxidase activities from S. media.
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Antivirales/aislamiento & purificación , Proliferación Celular/efectos de los fármacos , Herpesvirus Humano 2/efectos de los fármacos , Peroxidasas/metabolismo , Proteínas de Plantas/aislamiento & purificación , Stellaria/metabolismo , Secuencia de Aminoácidos , Animales , Antivirales/química , Antivirales/farmacología , Chlorocebus aethiops , Datos de Secuencia Molecular , Peroxidasas/química , Peroxidasas/aislamiento & purificación , Peroxidasas/farmacología , Proteínas de Plantas/química , Proteínas de Plantas/farmacología , Células VeroRESUMEN
American shad (Alosa sapidissima), introduced from the United States, has become one of the most expensive farmed fish in the aquatic product market of China. The shad reveals significant sexual dimorphism in growth and behaviors. For the study, five male-specific tags were identified in two-generation breeding populations of Alosa sapidissima and were verified by PCR amplification. Averages of 10,245,091 and 8,685,704 raw and enzyme reads were obtained by high-throughput sequencing of the 2b-RAD library, respectively. 301,022 unique tags were obtained from the sequences of twenty samples with sequencing depths of 0 to 500. Finally, 274,324 special tags and 29,327 SNPs were selected with a sequencing depth of 3 to 500. Eleven preliminary screening male-specific tags and three male heterogametic SNP loci were isolated. After verification by PCR amplification, five male-specific sequences of 27 bp located on chromosome 3 were screened out. Chromosome 3 could be assumed to be the sex chromosome of Alosa sapidissima. Sex-specific markers will provide invaluable and systematic animal germplasm resources to allow for the precise identification of neo-males for the all-female breeding of Alosa sapidissima in commercial aquaculture.
Asunto(s)
Peces , Secuenciación de Nucleótidos de Alto Rendimiento , Femenino , Masculino , Animales , Humanos , Acuicultura , China , Cromosomas Humanos Par 3RESUMEN
A multi-engine highly integrated microrobot, which is a Janus hemispherical shell structure composed of Pt and α-Fe2 O3 , is successfully developed. The microrobot can be efficiently driven and flexibly regulated by five stimuli, including an optical field, an acoustic field, magnetic field, an electric field, and chemical fuel. In addition, no matter which way it is driven by, the direction can be effectively controlled through the magnetic field regulation. Furthermore, this microrobot can also utilize magnetic or acoustic fields to achieve excellent aggregation control and swarm movement. Finally, this study demonstrates that the microrobots' propulsion can be effectively synergistically enhanced through the simultaneous action of two driving mechanisms, which can greatly improve the performance of the motor in applications, such as pollutant degradation. This multi-engine, highly integrated microrobot not only can adapt to more complex environments and has a wider application range, better application prospects, but also provides important ideas for designing future advanced micro/nanorobots.
RESUMEN
Liver fibrosis is a global health problem caused by chronic liver injury resulting from various factors. Hepatic stellate cells (HSCs) have been found to play a major role in liver fibrosis, and pathological stimuli lead to their transdifferentiation into myofibroblasts. Complex multidirectional interactions between HSCs, immune cells, and cytokines are also critical for the progression of liver fibrosis. Despite the advances in treatments for liver fibrosis, they do not meet the current medical needs. Exosomes are extracellular vesicles of 30-150 nm in diameter and are capable of intercellular transport of molecules such as lipids, proteins and nucleic acids. As an essential mediator of intercellular communication, exosomes are involved in the physiological and pathological processes of many diseases. In liver fibrosis, exosomes are involved in the pathogenesis mainly by regulating the activation of HSCs and the interaction between HSCs and immune cells. Serum-derived exosomes are promising biomarkers of liver fibrosis. Exosomes also have promising therapeutic potential in liver fibrosis. Exosomes derived from mesenchymal stem cells and other cells exhibit anti-liver fibrosis effects. Moreover, exosomes may serve as potential therapeutic targets for liver fibrosis and hold promise in becoming drug carriers for liver fibrosis treatment.