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Hemicellulose is one of the most important natural polysaccharides in nature. Hemicellulose from different sources varies in chemical composition and structure, which in turn affects the modification effects and industrial applications. Grain and oil by-products (GOBPs) are important raw materials for hemicellulose. This article reviews the modification methods of hemicellulose in GOBPs. The effects of chemical and physical modification methods on the properties of GOBP hemicellulose biomaterials are evaluated. The potential applications of modified GOBP hemicellulose are discussed, including its use in film production, hydrogel formation, three-dimensional (3D) printing materials, and adsorbents for environmental remediation. The limitations and future recommendations are also proposed to provide theoretical foundations and technical support for the efficient utilization of these by-products.
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Polyploidy is an important evolutionary force, yet epigenetic mechanisms, such as DNA methylation, that regulate genome-wide expression of duplicated genes remain largely unknown. Here, we use Tragopogon (Asteraceae) as a model system to discover patterns and temporal dynamics of DNA methylation in recently formed polyploids. The naturally occurring allotetraploid Tragopogon miscellus formed in the last 95-100 yr from parental diploids Tragopogon dubius and T. pratensis. We profiled the DNA methylomes of these three species using whole-genome bisulfite sequencing. Genome-wide methylation levels in T. miscellus were intermediate between its diploid parents. However, nonadditive CG and CHG methylation occurred in transposable elements (TEs), with variation among TE types. Most differentially methylated regions (DMRs) showed parental legacy, but some novel DMRs were detected in the polyploid. Differentially methylated genes (DMGs) were also identified and characterized. This study provides the first assessment of both overall and locus-specific patterns of DNA methylation in a recent natural allopolyploid and shows that novel methylation variants can be generated rapidly after polyploid formation. Together, these results demonstrate that mechanisms to regulate duplicate gene expression may arise soon after allopolyploid formation and that these mechanisms vary among genes.
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Asteraceae , Tragopogon , Tragopogon/genética , Asteraceae/genética , Metilação de DNA/genética , Poliploidia , Genoma de PlantaRESUMO
Background: Schizophrenia often occurs in youth, and psychosis risk syndrome (PRS) occurs before the onset of psychosis. Assessing the neuropsychological abnormalities of PRS individuals can help in early identification and active intervention of mental illness. Auditory P300 amplitude defect is an important manifestation of attention processing abnormality in PRS, but it is still unclear whether there are abnormalities in the attention processing of rhythmic compound tone stimuli in PRS individuals, and whether the P300 amplitude induced by these stimuli is specific to PRS individuals and related to their clinical outcomes. Methods: In total, 226 participants, including 122 patients with PRS, 51 patients with emotional disorders (ED), and 53 healthy controls (HC) were assessed. Baseline electroencephalography was recorded during the compound tone oddball task. The event-related potentials (ERPs) induced by rhythmic compound tone stimuli of two frequencies (20-Hz, 40-Hz) were measured. Almost all patients with PRS were followed up for 12 months and reclassified into four groups: PRS-conversion, PRS-symptomatic, PRS-emotional disorder, and PRS-complete remission. The differences in baseline ERPs were compared among the clinical outcome groups. Results: Regardless of the stimulation frequency, the average P300 amplitude were significantly higher in patients with PRS than in those with ED (p = 0.003, d = 0.48) and in HC (p = 0.002, d = 0.44) group. The average P300 amplitude of PRS-conversion group was significantly higher than that of the PRS-complete remission (p = 0.016, d = 0.72) and HC group (p = 0.001, d = 0.76), and the average P300 amplitude of PRS-symptomatic group was significantly higher than that of the HC group (p = 0.006, d = 0.48). Regardless of the groups (PRS, ED, HC) or the PRS clinical outcome groups, the average P300 amplitude induced by 20-Hz tone stimulation was significantly higher than that induced by 40-Hz stimulation (ps < 0.001, È 2 = 0.074-0.082). The average reaction times of PRS was significantly faster than that of ED (p = 0.01, d = 0.38), and the average reaction times of the participants to 20-Hz target stimulation was significantly faster than that to 40-Hz target stimulation (p < 0.001, d = 0.21). Conclusion: The auditory P300 amplitude induced by rhythmic compound tone stimuli is a specific electrophysiological manifestation of PRS, and the auditory P300 amplitude induced by compound tone stimuli shows promise as a putative prognostic biomarker for PRS clinical outcomes, including conversion to psychosis and clinical complete remission.
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Accurate and ultrasensitive evaluation of human epidermal growth factor receptor 2 (HER2) protein is key to early diagnosis and subtype differentiation of breast cancer. Single-cell analyses to reduce ineffective targeted therapies due to breast cancer heterogeneity and improve patient survival remain challenging. Herein, we reported a novel droplet microfluidic combined with an instant cation exchange signal amplification strategy for quantitative analysis of HER2 protein expression on single cells. In the 160 µm droplets produced by a tapered capillary bundle, abundant Immuno-CdS labeled on HER2-positive cells were replaced by Ag + to obtain Cd2+ that stimulated Rhod-5N fluorescence. This uniformly distributed and instantaneous fluorescence amplification strategy in droplets improves sensitivity and reduces signal fluctuation. Using HER2 modified PS microsphere to simulate single cells, we obtained a linear fitting of HER2-modified concentration and fluorescence intensity in microdroplets with the limit detection of 11.372 pg mL-1. Moreover, the relative standard deviation (RSD) was 4.2-fold lower than the traditional immunofluorescence technique (2.89% vs 12.21%). The HER2 protein on SK-BR-3 cells encapsulated in droplets was subsequently quantified, ranging from 9862.954 pg mL-1 and 205.26 pg mL-1, equivalent to 9.795 × 106 and 2.038 × 105 protein molecules. This detection system provides a universal platform for single-cell sensitive quantitative analysis and contributes to the evaluation of HER2-positive tumors.
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Neoplasias da Mama , Receptor ErbB-2 , Humanos , Feminino , Receptor ErbB-2/metabolismo , Imunofluorescência , Neoplasias da Mama/diagnósticoRESUMO
Background: Low grade of sterile inflammation plays detrimental roles in the progression of diabetic kidney disease (DKD). Sappanone A (SA), a kind of homoisoflavanone isolated from the heartwood of Caesalpinia sappan, exerts anti-inflammatory effects in acute kidney injury. However, whether SA has beneficial effects on diabetic kidney disease remains further exploration. Methods and Results: In the present study, uninephrectomized male mice were treated with Streptozotocin (STZ, 50 mg/kg) for five consecutive days to induce diabetes. Next, the diabetic mice were administered orally with SA (10, 20, or 30 mg/kg) or vehicle once per day. Our results showed that STZ treatment significantly enhanced damage in the kidney, as indicated by an increased ratio of kidney weight/body weight, elevated serum creatinine and blood urea nitrogen (BUN), as well as increased 24-h urinary protein excretion, whereas SA-treated mice exhibited a markedly amelioration in these kidney damages. Furthermore, SA attenuated the pathological changes, alleviated fibrotic molecules transforming growth factor-ß1 (TGF-ß1) and Collagen-IV (Col-IV) production, decreased inflammatory cytokines interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) expression in STZ-treated mice. Similarly, in glomerular mesangial cells, SA pretreatment decreased high glucose (HG)-induced proliferation, inflammatory cytokines excretion, and fibrotic molecules expression. Mechanistically, SA decreased the expression of nuclear factor kappa B (NF-κB) and restored the expression of total NF-κB inhibitor alpha (IκBα) both in vivo and in vitro. Conclusion: Our data suggest that SA may prevent diabetes-induced kidney inflammation and fibrosis by inhibiting the NF-κB pathway. Hence, SA can be potential and specific therapeutic value in DKD.
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Respiratory syncytial virus (RSV) is a major cause of acute lower respiratory tract infection in infants and children. The present study aimed to investigate the effects of miR-146a on RSV replication and the related mechanisms. MATERIAL AND METHODS: We pretreated A549 and HEp-2 cells and young rats with miR-146a mimic before infection with RSV. The expressions of miR-146a and RSV-F mRNA in cells and lung tissues were detected by RT-qPCR, and production of IL-1ß, IL-6, IL-18, and TNF-α in bronchial alveolar lavage fluid (BALF) were determined by ELISA. The expression level of TRAF-6 and activation of the JNK/ERK/MAPK/NF-κB signaling pathway was detected by Western blotting. RESULTS: RSV infection significantly reduced miR-146a levels in both A549 and HEp-2 cells and rat lung tissues. RSV infection resulted in accelerated growth, increased release of inflammatory cytokines, increased expression of TRAF-6, and activation of the JNK pathway in cells, and the lung inflammatory infiltration and the pathological score increased in rats. Overexpression of miR-146a targeted down-regulation of TRAF-6 expression and JNK/ERK/MAPK/NF-κB pathway induced by RSV infection, reduced the production of inflammatory cytokines IL-1ß, IL-6 and TNF-α, and alleviate lung injury in young rats. We got similar results in both A549 and HEp-2 cell experiments. CONCLUSION: MiR-146a alleviates lung injury caused by RSV infection in young rats by targeting TRAF-6 and regulating JNK/ERK/MAPK signaling pathways.
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Lesão Pulmonar , MicroRNAs/genética , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Animais , Citocinas/metabolismo , Interleucina-6/metabolismo , Lesão Pulmonar/etiologia , Sistema de Sinalização das MAP Quinases , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Ratos , Vírus Sincicial Respiratório Humano/genética , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Polyploidy is an important evolutionary mechanism and is prevalent among land plants. Most polyploid species examined have multiple origins, which provide genetic diversity and may enhance the success of polyploids. In some polyploids, recurrent origins can result from reciprocal crosses between the same diploid progenitors. Although great progress has been made in understanding the genetic consequences of polyploidy, the genetic implications of reciprocal polyploidization remain poorly understood, especially in natural polyploids. Tragopogon (Asteraceae) has become an evolutionary model system for studies of recent and recurrent polyploidy. Allotetraploid T. miscellus has formed reciprocally in nature with resultant distinctive floral and inflorescence morphologies (i.e., short- vs. long-liguled forms). In this study, we performed comparative inflorescence transcriptome analyses of reciprocally formed T. miscellus and its diploid parents, T. dubius and T. pratensis. In both forms of T. miscellus, homeolog expression of â¼70% of the loci showed vertical transmission of the parental expression patterns (i.e., parental legacy), and â¼20% of the loci showed biased homeolog expression, which was unbalanced toward T. pratensis. However, 17.9% of orthologous pairs showed different homeolog expression patterns between the two forms of T. miscellus. No clear effect of cytonuclear interaction on biased expression of the maternal homeolog was found. In terms of the total expression level of the homeologs studied, 22.6% and 16.2% of the loci displayed non-additive expression in short- and long-liguled T. miscellus, respectively. Unbalanced expression level dominance toward T. pratensis was observed in both forms of T. miscellus. Significantly, genes annotated as being involved in pectin catabolic processes were highly expressed in long-liguled T. miscellus relative to the short-liguled form, and the majority of these differentially expressed genes were transgressively down-regulated in short-liguled T. miscellus. Given the known role of these genes in cell expansion, they may play a role in the differing floral and inflorescence morphologies of the two forms. In summary, the overall inflorescence transcriptome profiles are highly similar between reciprocal origins of T. miscellus. However, the dynamic homeolog-specific expression and non-additive expression patterns observed in T. miscellus emphasize the importance of reciprocal origins in promoting the genetic diversity of polyploids.
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Early diagnosis of cancer biomarkers is the key to guiding treatments and improving the survival rate of patients. Herein, we report a novel surface-enhanced resonance Raman scattering (SERRS)-based lateral flow immunoassay (LFIA) for quantitative and ultra-sensitive analysis of alpha-fetoprotein (AFP). Gold nanorods (AuNRs) were fabricated to be in resonance with 785 nm laser excitation, that is, the excitation level that can maximize SERRS activity. The AuNRs were modified with 5,5'-dithiobis(2-nitrobenzoic acid), bovine serum albumin (BSA), and AFP detection antibody successively as the SERRS nanotags for the LFIA system. Modification of the BSA layer guaranteed good stability and biocompatibility of the SERRS nanotags in complex samples. The SERRS-LFIA strip for AFP detection showed a low detection limit of 9.2 pg mL-1 and a broad detection range from 10 pg mL-1 to 500 ng mL-1. By comparison, the detection limit of the proposed assay is about 100 and 10 times lower than those of the Au nanoparticle-based SERS-strip and conventional enzyme-linked immunosorbent assay, respectively. Moreover, the potential clinical applications of the assay were evaluated by detecting 10 actual serum samples. Results showed 100% accuracy based on the clinical tests.
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The present study aimed to investigate the effect of avicularin on rheumatoid arthritis (RA) in vitro, and additionally explore the molecular mechanism. To perform this investigation, an in vitro model of RA was established by treatment of the human RA synovial MH7A cell line with tumor necrosis factor-α (TNF-α). MH7A cells were then treated with various concentrations (10, 30, 100 and 300 µM) of avicularin. Then, the levels of inflammatory factors [interleukin (IL)-1ß, IL-6, IL-8, matrix metalloproteinase (MMP)-1 and MMP-13] were measured by ELISA. Cell viability and apoptosis were detected using an MTT assay and flow cytometry, respectively. In addition, the expression levels of genes and proteins were determined reverse transcription quantitative polymerase chain reaction and western blot analysis. The results of the present study indicated that avicularin significantly decreased the levels of inflammatory factors (IL-1ß, IL-6, IL-8, MMP-1 and MMP-13), previously increased by TNF-α, in a dose-dependent manner. Concurrently, avicularin inhibited the mRNA and protein expression levels of iNOS and COX-2 increased by TNF-α. It was also identified that TNF-α administration significantly promoted MH7A cell viability and inhibited cell apoptosis, and avicularin treatment dose-dependently inhibited MH7A cell viability and induced cell apoptosis. In addition, these data suggested that avicularin prevented the activation of the mitogen-activated protein kinase kinase (MEK)/nuclear factor kappa light-chain-enhancer of activated B-cells (NF-κB) pathway activated by TNF-α. Taken together, these results demonstrated that avicularin may inhibit the inflammatory response, prevent cell viability and induce apoptosis in human RA synovial cells through preventing the activation of the MEK/NF-κB pathway.
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Microsatellites, or simple sequence repeats (SSRs), have long played a major role in genetic studies due to their typically high polymorphism. They have diverse applications, including genome mapping, forensics, ascertaining parentage, population and conservation genetics, identification of the parentage of polyploids, and phylogeography. We compare SSRs and newer methods, such as genotyping by sequencing (GBS) and restriction site associated DNA sequencing (RAD-Seq), and offer recommendations for researchers considering which genetic markers to use. We also review the variety of techniques currently used for identifying microsatellite loci and developing primers, with a particular focus on those that make use of next-generation sequencing (NGS). Additionally, we review software for microsatellite development and report on an experiment to assess the utility of currently available software for SSR development. Finally, we discuss the future of microsatellites and make recommendations for researchers preparing to use microsatellites. We argue that microsatellites still have an important place in the genomic age as they remain effective and cost-efficient markers.
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Allopolyploidy involves hybridization and duplication of divergent parental genomes and provides new avenues for gene expression. The expression levels of duplicated genes in polyploids can show deviation from parental additivity (the arithmetic average of the parental expression levels). Nonadditive expression has been widely observed in diverse polyploids and comprises at least three possible scenarios: (a) The total gene expression level in a polyploid is similar to that of one of its parents (expression-level dominance); (b) total gene expression is lower or higher than in both parents (transgressive expression); and (c) the relative contribution of the parental copies (homeologs) to the total gene expression is unequal (homeolog expression bias). Several factors may result in expression nonadditivity in polyploids, including maternal-paternal influence, gene dosage balance, cis- and/or trans-regulatory networks, and epigenetic regulation. As our understanding of nonadditive gene expression in polyploids remains limited, a new generation of investigators should explore additional phenomena (i.e., alternative splicing) and use other high-throughput "omics" technologies to measure the impact of nonadditive expression on phenotype, proteome, and metabolome.
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Epigênese Genética , Regulação da Expressão Gênica de Plantas/genética , Hibridização Genética , Poliploidia , Processamento Alternativo/genética , Arabidopsis/genética , Dosagem de Genes , Genoma de PlantaRESUMO
Nearly four decades ago, Roose & Gottlieb (Roose & Gottlieb 1976 Evolution 30, 818-830. (doi:10.2307/2407821)) showed that the recently derived allotetraploids Tragopogon mirus and T. miscellus combined the allozyme profiles of their diploid parents (T. dubius and T. porrifolius, and T. dubius and T. pratensis, respectively). This classic paper addressed the link between genotype and biochemical phenotype and documented enzyme additivity in allopolyploids. Perhaps more important than their model of additivity, however, was their demonstration of novelty at the biochemical level. Enzyme multiplicity-the production of novel enzyme forms in the allopolyploids-can provide an extensive array of polymorphism for a polyploid individual and may explain, for example, the expanded ranges of polyploids relative to their diploid progenitors. In this paper, we extend the concept of evolutionary novelty in allopolyploids to a range of genetic and ecological features. We observe that the dynamic nature of polyploid genomes-with alterations in gene content, gene number, gene arrangement, gene expression and transposon activity-may generate sufficient novelty that every individual in a polyploid population or species may be unique. Whereas certain combinations of these features will undoubtedly be maladaptive, some unique combinations of newly generated variation may provide tremendous evolutionary potential and adaptive capabilities.
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Adaptação Biológica/genética , Evolução Biológica , Enzimas/genética , Variação Genética , Modelos Genéticos , Fenótipo , Poliploidia , Tragopogon/genéticaRESUMO
OBJECTIVE: To explore the effect of transforming growth factor beta1 (TGF-beta1) on epithelial-mesenchymal transition in rat peritoneal mesothelial cells(RPMCs) and its mechanism. METHODS: Primary peritoneal mesothelial cells of SP rats were cultured in vitro. After synchronization for 24 h, RPMCs were randomly divided into 2 groups: Group A (control), Group B (TGF-beta1, 10 mug/L). RPMCs were stimulated by 10 mug/L TGF-beta1 for different time. The mRNA and protein expression levels of E-cadherin, alpha-smooth muscle actin (alpha-SMA) and collagen I were measured by RT-PCR and Western blot, respectively. The protein expression level of total RhoA was measured by Western blot. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, and assessed by Western blot. RESULTS: TGF-beta1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of alpha-SMA and CollagenI. TGF-beta1 stimulation elicited a robust increase in RhoA activity in a time-dependent manner. RhoA activity peaked at 1 h. CONCLUSION: RPMCs can be transdifferentiated into myofibroblast under the effect of TGF-beta1,and the mechanism may be related to the activation of RhoA associated signal pathway.