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Passion fruit (Passiflora edulis) possesses a complex aroma and is widely grown in tropical and subtropical areas. Here, we conducted the de novo assembly, annotation, and comparison of PPF (P. edulis Sims) and YPF (P. edulis f. flavicarpa) reference genomes using PacBio, Illumina, and Hi-C technologies. Notably, we discovered evidence of recent whole-genome duplication events in P. edulis genomes. Comparative analysis revealed 7.6â¼8.1 million single nucleotide polymorphisms, 1 million insertions/deletions, and over 142â Mb presence/absence variations among different P. edulis genomes. During the ripening of yellow passion fruit, metabolites related to flavor, aroma, and color were substantially accumulated or changed. Through joint analysis of genomic variations, differentially expressed genes, and accumulated metabolites, we explored candidate genes associated with flavor, aroma, and color distinctions. Flavonoid biosynthesis pathways, anthocyanin biosynthesis pathways, and related metabolites are pivotal factors affecting the coloration of passion fruit, and terpenoid metabolites accumulated more in PPF. Finally, by heterologous expression in yeast (Saccharomyces cerevisiae), we functionally characterized 12 terpene synthases. Our findings revealed that certain TPS homologs in both YPF and PPF varieties produce identical terpene products, while others yield distinct compounds or even lose their functionality. These discoveries revealed the genetic and metabolic basis of unique characteristics in aroma and flavor between the 2 passion fruit varieties. This study provides resources for better understanding the genome architecture and accelerating genetic improvement of passion fruits.
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Frutas , Passiflora , Frutas/genética , Odorantes , Passiflora/genética , Passiflora/metabolismo , Multiómica , Terpenos/metabolismoRESUMEN
T-cell receptor (TCR)-engineered T cells can precisely recognize a broad repertoire of targets derived from both intracellular and surface proteins of tumor cells. TCR-T adoptive cell therapy has shown safety and promising efficacy in solid tumor immunotherapy. However, antigen-specific functional TCR screening is time-consuming and expensive, which limits its application clinically. Here, we developed a novel integrated antigen-TCR screening platform based on droplet microfluidic technology, enabling high-throughput peptide-major histocompatibility complex (pMHC)-to-TCR paired screening with a high sensitivity and low background signal. We introduced DNA barcoding technology to label peptide antigen candidate-loaded antigen-presenting cells and Jurkat reporter cells to check the specificity of pMHC-TCR candidates. Coupled with the next-generation sequencing pipeline, interpretation of the DNA barcodes and the gene expression level of the Jurkat T-cell activation pathway provided a clear peptide-MHC-TCR recognition relationship. Our proof-of-principle study demonstrates that the platform could achieve pMHC-TCR paired high-throughput screening, which is expected to be used in the cross-reactivity and off-target high-throughput paired testing of candidate pMHC-TCRs in clinical applications.
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Ensayos Analíticos de Alto Rendimiento , Microfluídica , Humanos , Receptores de Antígenos de Linfocitos T/genética , Receptores de Antígenos de Linfocitos T/metabolismo , Antígenos , Péptidos/metabolismoRESUMEN
Electrochemical aptasensing systems have been developed for screening low-abundance disease-related proteins, but most of them involve multiple washings and multi-step separation during measurements, and thus are disadvantageous for routine use. In this work, an innovative and simple electrochemical aptasensing platform was designed for the voltammetric detection of prostate-specific antigen (PSA) in biological fluids without any washing and separation steps. This system mainly included a PSA-specific aptamer, a DNA walker and two hairpin DNA probes (i.e., thiolated hairpin DNA1 and ferrocene-labeled hairpin DNA2). Introduction of target PSA caused the release of the DNA walker from a partially complementary aptamer/DNA walker hybridization strand. The dissociated DNA walker opened the immobilized hairpin DNA1 on the electrode, accompanying subsequent displacement reaction with hairpin DNA2, thus resulting in the DNA walker step-by-step reaction with numerous hairpin DNA1 probes on the sensing interface. In this case, numerous ferrocene molecules were close to the electrode to amplify the voltammetric signal within the applied potentials. All reactions and electrochemical measurements including the target/aptamer reaction and hybridization chain reaction were implemented in the same detection cell. Under optimal conditions, the fabricated electrochemical aptasensor gave good voltammetric responses relative to the PSA concentrations within the range of 0.001-10 ng mL-1 at an ultralow detection limit of 0.67 pg mL-1. A good reproducibility with batch-to-batch errors was acquired for target PSA down to 11.5%. Non-target analytes did not interfere with the voltammetric signals of the electrochemical aptasensors. Meanwhile, 15 human serum specimens were measured with electrochemical aptasensors, and displayed well-matched results in comparison with the referenced human PSA enzyme-linked immunosorbant assay (ELISA) method. Significantly, this method provides a new horizon for the quantitative monitoring of low-concentration biomarkers or nucleic acids.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , ADN/química , Sondas de ADN/genética , Técnicas Electroquímicas/métodos , Oro/química , Humanos , Límite de Detección , Masculino , Metalocenos , Antígeno Prostático Específico , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: It is of utmost importance to investigate novel therapies for cancer, as it is a major cause of death. In recent years, immunotherapies, especially those against immune checkpoints, have been developed and brought significant improvement in cancer management. However, on the other hand, immune checkpoints blockade (ICB) by monoclonal antiboties may cause common and severe adverse reactions (ADRs), the cause of which remains largely undetermined. We hypothesize that ICB-agents may induce adverse reactions through off-target protein interactions, similar to the ADR-causing off-target effects of small molecules. In this study, we propose a hybrid phenotype mining approach which integrates molecular level information and provides new mechanistic insights for ICB-associated ADRs. METHODS: We trained a conditional random fields model on the TAC 2017 benchmark training data, then used it to extract all drug-centric phenotypes for the five anti-PD-1/PD-L1 drugs from the drug labels of the DailyMed database. Proteins with structure similar to the drugs were obtained by using BlastP, and the gene targets of drugs were obtained from the STRING database. The target-centric phenotypes were extracted from the human phenotype ontology database. Finally, a screening module was designed to investigate off-target proteins, by making use of gene ontology analysis and pathway analysis. RESULTS: Eventually, through the cross-analysis of the drug and target gene phenotypes, the off-target effect caused by the mutation of gene BTK was found, and the candidate side-effect off-target site was analyzed. CONCLUSIONS: This research provided a hybrid method of biomedical natural language processing and bioinformatics to investigate the off-target-based mechanism of ICB treatment. The method can also be applied for the investigation of ADRs related to other large molecule drugs.
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Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Neoplasias , Humanos , Inmunoterapia/efectos adversos , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Fenotipo , ProteínasRESUMEN
The environmental information disclosure system is regarded as an important environmental management tool for supervising the environmental behavior of listed companies in China. To explore what drives the environmental information disclosure, a configurational analysis targeted at 264 China's key pollutant-discharge listed companies was studied by fuzzy-set qualitative comparative analysis (fs/QCA). The results showed that there were three types of paths to drive the environmental information disclosure: "Strict regulation - low resources - weak capability", "Strict regulation - low resources - strong capability" and "Strict regulation - high resources - strong capability", and that strict environmental regulation played a greatly significant role for promoting the environmental information disclosure level. The research provides a theoretical basis for the effectiveness of environmental regulation on environmental information disclosure, and it charts the future direction for environmental management in China.
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Contaminantes Ambientales , China , RevelaciónRESUMEN
A germinating seedling undergoes skotomorphogenesis to emerge from the soil and reach for light. During this phase, the cotyledons are closed, and the hypocotyl elongates. Upon exposure to light, the seedling rapidly switches to photomorphogenesis by opening its cotyledons and suppressing hypocotyl elongation. The E3 ubiquitin ligase CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) is critical for maintaining skotomorphogenesis. Here, we report that jasmonate (JA) suppresses hypocotyl elongation and stimulates cotyledon opening in etiolated seedlings, partially phenocopying cop1 mutants in the dark. We also find that JA stabilizes several COP1-targeted transcription factors in a COP1-dependent manner. RNA-seq analysis further defines a JA-light co-modulated and cop1-dependent transcriptome, which is enriched for auxin-responsive genes and genes participating in cell wall modification. JA suppresses COP1 activity through at least two distinct mechanisms: decreasing COP1 protein accumulation in the nucleus; and reducing the physical interaction between COP1 and its activator, SUPPRESSOR OF PHYTOCHROME A-105 1 (SPA1). Our work reveals that JA suppresses COP1 activity to stabilize COP1 targets, thereby inhibiting hypocotyl elongation and stimulating cotyledon unfolding in etiolated Arabidopsis seedlings.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Ciclopentanos/farmacología , Hipocótilo/crecimiento & desarrollo , Hipocótilo/metabolismo , Oxilipinas/farmacología , Plantones/crecimiento & desarrollo , Plantones/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Arabidopsis/efectos de los fármacos , Proteínas de Arabidopsis/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Cotiledón/efectos de los fármacos , Cotiledón/crecimiento & desarrollo , Cotiledón/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/genética , Hipocótilo/efectos de los fármacos , Plantones/efectos de los fármacos , Ubiquitina-Proteína Ligasas/genéticaRESUMEN
Bananas (Musa spp.) are monocotyledonous plants with high genetic diversity in the Musaceae family that are cultivated mainly in tropical and subtropical countries. The fruits are a popular food, and the plants themselves have diverse uses. Four genetic groups (genomes) are thought to have contributed to current banana cultivars: Musa acuminata (A genome), Musa balbisiana (B genome), Musa schizocarpa (S genome), and species of the Australimusa section (T genome). However, the T genome has not been effectively explored. Here, we present the high-quality TT genomes of two representative accessions, Abaca (Musa textilis), with high-quality natural fiber, and Utafun (Musa troglodytarum, Fe'i group), with abundant ß-carotene. Both the Abaca and Utafun assemblies comprise 10 pseudochromosomes, and their total genome sizes are 613 Mb and 619 Mb, respectively. Comparative genome analysis revealed that the larger size of the T genome is likely attributable to rapid expansion and slow removal of transposons. Compared with those of Musa AA or BB accessions or sisal (Agava sisalana), Abaca fibers exhibit superior mechanical properties, mainly because of their thicker cell walls with a higher content of cellulose, lignin, and hemicellulose. Expression of MusaCesA cellulose synthesis genes peaks earlier in Abaca than in AA or BB accessions during plant development, potentially leading to earlier cellulose accumulation during secondary cell wall formation. The Abaca-specific expressed gene MusaMYB26, which is directly regulated by MusaMYB61, may be an important regulator that promotes precocious expression of secondary cell wall MusaCesAs. Furthermore, MusaWRKY2 and MusaNAC68, which appear to be involved in regulating expression of MusaLAC and MusaCAD, may at least partially explain the high accumulation of lignin in Abaca. This work contributes to a better understanding of banana domestication and the diverse genetic resources in the Musaceae family, thus providing resources for Musa genetic improvement.
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Musa , Musa/genética , Genoma de Planta , LigninaRESUMEN
Bananas (Musa spp.) are one of the world's most important fruit crops and play a vital role in food security for many developing countries. Most banana cultivars are triploids derived from inter- and intraspecific hybridizations between the wild diploid ancestor species Musa acuminate (AA) and M. balbisiana (BB). We report two haplotype-resolved genome assemblies of the representative AAB-cultivated types, Plantain and Silk, and precisely characterize ancestral contributions by examining ancestry mosaics across the genome. Widespread asymmetric evolution is observed in their subgenomes, which can be linked to frequent homologous exchange events. We reveal the genetic makeup of triploid banana cultivars and verify that subgenome B is a rich source of disease resistance genes. Only 58.5% and 59.4% of Plantain and Silk genes, respectively, are present in all three haplotypes, with >50% of genes being differentially expressed alleles in different subgenomes. We observed that the number of upregulated genes in Plantain is significantly higher than that in Silk at one-week post-inoculation with Fusarium wilt tropical race 4 (Foc TR4), which confirms that Plantain can initiate defense responses faster than Silk. Additionally, we compared genomic and transcriptomic differences among the genes related to carotenoid synthesis and starch metabolism between Plantain and Silk. Our study provides resources for better understanding the genomic architecture of cultivated bananas and has important implications for Musa genetics and breeding.
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Fusarium , Musa , Musa/genética , Fusarium/genética , Haplotipos , Perfilación de la Expresión Génica , TranscriptomaRESUMEN
OBJECTIVE: To analyze the prevalence and risk factors of multiple musculoskeletal disorders (MSDs) in auto workers and the associations between MSDs at different sites. METHODS: A cross-sectional survey was conducted in 3998 workers, who were selected from a Chinese auto corporation by cluster random sampling, using the revised Nordic MSDs standard questionnaire; 3800 completed questionnaires were returned. Multinomial logistic regression analysis was performed to assess the risk factors for multiple MSDs. The logbinomial model was used to calculate the prevalence ratios (PRs) of MSDs at different sites and evaluate the associations between MSDs at different sites. RESULTS: Of the 3800 subjects, 2452 (64.5%) had MSDs at two or more sites, and 469 (12.3%) had MSDs at one site. The PRs varied from 1.5 to 6.7, with significant differences among different sites (P < 0.01). Relatively close associations were found between the MSDs at neck and shoulders, back and shoulders/waist, elbows and wrists/hands, waist and neck, wrists/hands and waist, hip and waist, knees and waist, and ankles/feet and elbows. The multinomial logistic regression analysis indicated that the highest risk factor for MSDs was poor posture, including often working in an uncomfortable posture, neck bending forward, and neck twisting (ORs = 3.39, 1.93, and 1.38), followed by labor organization, in which break between tasks could decrease the risk of MSDs at three or more sites to 31%, staff shortage, which could increase the risk of MSDs by 75%, and pushing and pulling heavy objects (> 20 kg) (OR = 1.76). CONCLUSION: Most auto workers with MSDs have multiple sites affected, and there are high associations between the MSDs at different sites. The major risk factors for multiple MSDs in auto workers include poor posture, labor organization, and heavy physical labor.
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Enfermedades Musculoesqueléticas/epidemiología , Enfermedades Profesionales/epidemiología , Adulto , Automóviles , Estudios Transversales , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Enfermedades Musculoesqueléticas/etiología , Enfermedades Profesionales/etiología , Factores de Riesgo , Encuestas y CuestionariosRESUMEN
Triazole antifungal pesticides work by inhibiting the activity of lanosterol-14-α-demethylase, a member of cytochrome P450 enzymes (CYPs), but this effect is non-specific. Bile acids (BAs) are important physical surfactants for lipids absorption in intestine, and synthesized by CYPs 7A1/27A1. Thus, we presume that triazole exposure might influence the therapeutic effect or safety of oral medication through disturbing the BAs pool, even though the exposure is under an acceptable daily intake (ADI) dose. Short- and long-term of ADI dose tebuconazole (TEB) exposure animal models were established through various routes, and statins with different hydrophilic and lipophilic properties were gavaged. It exhibited that the activity of CYP7A1/27A1 was indeed inhibited but the expression was up-regulated, the BAs pool was changed either the content and the composition, and the absorption behavior of statins with strong and medium degree of lipid-solubility were significantly changed. A series of experiments performed on models of intestinal mucus, Caco-2 cell monolayer and Caco-2/HT29 co-culture system revealed that the TEB-exposure induced BAs disturbance made impacts on drug absorption in many aspects, including drug solubility and the structure of intestinal barriers. This study suggests us to be more alert about the hazard of pesticides residues for elderly and chronically ill groups.
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Inhibidores de Hidroximetilglutaril-CoA Reductasas , Plaguicidas , Humanos , Animales , Ácidos y Sales Biliares , Células CACO-2 , Nivel sin Efectos Adversos ObservadosRESUMEN
Translational regulation is a critical step in the process of gene expression and governs the synthesis of proteins from mRNAs. Many studies have revealed translational regulation in plants in response to various environmental stimuli. However, there have been no studies documenting the comprehensive landscape of translational regulation and allele-specific translational efficiency in multiple plant tissues, especially those of rice, a main staple crop that feeds nearly half of the world's population. Here we used RNA sequencing and ribosome profiling data to analyze the transcriptome and translatome of an elite hybrid rice, Shanyou 63 (SY63), and its parental varieties Zhenshan 97 and Minghui 63. The results revealed that gene expression patterns varied more among tissues than among varieties at the transcriptional and translational levels. We identified 3392 upstream open reading frames (uORFs), and the uORF-containing genes were enriched in transcription factors. Only 668 of 13 492 long non-coding RNAs could be translated into peptides. Finally, we discovered numerous genes with allele-specific translational efficiency in SY63 and demonstrated that some cis-regulatory elements may contribute to allelic divergence in translational efficiency. Overall, these findings may improve our understanding of translational regulation in rice and provide information for molecular breeding research.
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Oryza , Biosíntesis de Proteínas , Biosíntesis de Proteínas/genética , Ribosomas/genética , Oryza/genética , Perfilado de Ribosomas , AlelosRESUMEN
BACKGROUND: Abnormalities in the mitochondrial energy metabolism pathways are closely related to the occurrence and development of many cancers. Furthermore, abnormal genes in mitochondrial energy metabolism pathways may be novel targets and biomarkers for the diagnosis and treatment of osteosarcoma. In this study, we aimed to establish a mitochondrial energy metabolism-related gene signature for osteosarcoma prognosis. METHODS: We first obtained differentially expressed genes based on the metastatic status of 84 patients with osteosarcoma from the TARGET database. After Venn analysis of differentially expressed genes and mitochondrial energy metabolism pathway-related genes (MMRGs), 2 key genes were obtained using univariate Cox regression and least absolute shrinkage and selection operator (LASSO) regression analysis. Next, we used these 2 genes to establish a prognostic signature. Subsequent analyses elucidated the correlation between these 2 key genes with clinical features and 28 types of immune cells. Pathway changes in osteosarcoma pathogenesis under different metastatic states were clarified using gene set enrichment analysis (GSEA) of differentially expressed genes. RESULTS: A gene signature composed of 2 key prognosis-related genes (KCNJ5 and PFKFB2) was identified. A risk score was calculated based on the gene signature, which divided osteosarcoma patients into low- or high-risk groups that showed good and poor prognosis, respectively. High expression of these 2 key genes is associated with low-risk group in patients with osteosarcoma. We constructed an accurate nomogram to help clinicians assess the survival time of patients with osteosarcoma. The results of immune cell infiltration level showed that the high-risk group had lower levels of immune cell infiltration. GSEA revealed changes in immune regulation and hypoxia stress pathways in osteosarcoma under different metastatic states. CONCLUSION: Our study identified an excellent gene signature that could be helpful in improving the prognosis of patients with osteosarcoma.
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Neoplasias Óseas , Osteosarcoma , Humanos , Pronóstico , Osteosarcoma/genética , Mitocondrias/genética , Metabolismo Energético/genética , Neoplasias Óseas/genética , Microambiente Tumoral/genética , Canales de Potasio Rectificados Internamente Asociados a la Proteína G , Fosfofructoquinasa-2RESUMEN
Myelodysplastic syndrome (MDS) consists of a group of hematologic tumors that are derived from the clonal proliferation of hematopoietic stem cells, featuring abnormal hematopoietic cell development and ineffective hematopoiesis. Animal models are an important scientific research platform that has been widely applied in the research of human diseases, especially tumors. Animal models with MDS can simulate characteristic human genetic variations and tumor phenotypes. They also provide a reliable platform for the exploration of the pathogenesis and diagnostic markers of MDS as well as for a drug efficacy evaluation. This paper reviews the research status of three animal models and a new spontaneous mouse model with MDS.
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Neoplasias Hematológicas , Síndromes Mielodisplásicos , Animales , Ratones , Humanos , Síndromes Mielodisplásicos/genética , Células Madre Hematopoyéticas/patología , Modelos Animales de Enfermedad , HematopoyesisRESUMEN
BACKGROUND: Henoch-Schonlein purpura (HSP) is a common capillary allergic bleeding disease. To explore the variation of pyroptosis-related inflammatory factors level in the peripheral blood of patients with HSP. METHODS: A total of 87 HSP patients treated in our hospital from June 2020 to March 2021 were selected and divided into the renal impairment group (n=29) and the non-renal impairment group (n=58) according to the presence of hematuria and proteinuria. A total of 50 healthy individuals from the hospital were selected as the control group. The renal impairment and non-renal impairment groups were treated with a regular regimen of compound glycyrrhizin tablets and glucocorticoids, respectively. Serum interleukin (IL)-18, IL-1ß, and peripheral caspase-1-positive cells were compared pre- and post-treatment among the three groups. RESULTS: The pre-treatment serum IL-1ß levels in the renal impairment and non-renal impairment groups were significantly higher than that in the control group (P<0.01). After treatment, the IL-1ß level in the non-renal impairment group was not significantly different from that in the control group (P>0.05). However, the IL-1ß level in the renal impairment group post-treatment was significantly higher than that in the other two groups (P<0.01). The positive rate of caspase-1 expression in peripheral blood before treatment in the renal impairment group and non-renal impairment group was significantly higher than that in the control group (P<0.01). After treatment, the positive rate of caspase-1 expression in the non-renal impairment group was comparable to that in the control group (P>0.05), whereas the rate in the renal impairment group was significantly higher than that in the other two groups (P<0.01). After treatment, the serum IL-1ß levels and caspase-1 positive rate in HSP patients who were responsive to treatment (as assessed by hematuria or proteinuria levels after treatment) were lower than that in patients who were unresponsive to treatment P<0.001), but not significantly different to the control group (P>0.05). CONCLUSIONS: The levels of serum IL-1ß and caspase-1 changed in response to alterations in the disease condition and treatment response in HSP patients, which suggested that pyroptosis-related inflammatory factors may have potential application value in predicting disease progression and efficacy of hormone therapy.
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Vasculitis por IgA , Glucocorticoides , Humanos , Vasculitis por IgA/tratamiento farmacológico , PiroptosisRESUMEN
Temozolomide (TMZ), one of the few effective drugs used during adjuvant therapy, could effectively prolong the overall survival (OS) of glioma patients. In our previous study, the mRNA level of G Protein Subunit Alpha 13 (GNA13) was found to be inversely correlated with OS and was therefore identified as a potential biomarker for the prognosis of glioma. Henceforth, this study aims to identify the molecular mechanism of GNA13 in enhancing TMZ sensitization through bioinformatic analyses of GSE80729 and GSE43452 and other experiments. In glioma, overexpression of GNA13 downregulated PRKACA, which is a subunit of PKA, hence reducing phosphorylated RELA and MGMT. Since p-RELA and MGMT were proven to be closely associated with TMZ resistance, we therefore investigated whether thetwo signaling pathways, "GNA13/PRKACA/p-RELA", and "GNA13/PRKACA/MGMT", were involved in the molecular mechanism of GNA13 in TMZ sensitization. Our conclusion was that, GNA13 overexpression in glioma cells were more sensitive in TMZ treatment.
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Objective: To investigate the dynamic regulation of self-assembled aggregations (SAA) in Coptidis Rhizoma decoction on the permeability of intestinal tissue and the mechanism underlying. Methods: The effects of SAA on berberine (Ber) absorption were respectively analyzed in an in situ intestinal perfusion model and in an Ussing Chamber jejunum model with or without Peyer's patches (PPs). The expression levels of ZO-1, Occludin and Claudin-1 were detected by immunofluorescence to evaluate the tight junction (TJ) between intestinal epithelium cells. The expression levels of T-box-containing protein expressed in T cells, signal transducers and activators of tranion-6, retinoic acid receptor-related orphan receptor γt and forkhead box P3 in PPs were detected by the reverse transcription-polymerase chain reaction and the secretions of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-ß (TGF-ß) in PPs were evaluated by immunohistochemistry, to reflect the differentiation of T lymphocyte in PPs to helper T (Th) cell 1, Th2, Th17 and regulatory T (Treg) cell. To confirm the correlation between SAA in Coptidis Rhizoma decoction, PPs-associated immunity and intestinal epithelium permeability, SAA were administrated on an Ussing Chamber jejunum model with immunosuppressed PPs and evaluated its influences on intestinal tissue permeability and TJ proteins expression. Results: SAA in Coptidis Rhizoma decoction could dose-dependently promote Ber absorption in jejunum segment, with the participation of PPs. The dose-dependent and dynamical regulations of SAA on permeability of intestinal tissue and TJ proteins expression level between intestinal epithelium cells occurred along with the dynamically changed T lymphocyte differentiation and immune effectors secretion in PPs. The administration of SAA on immunosuppressed PPs exhibited dose-dependent PPs activation, inducing dynamic promotion on intestinal tissue permeability and inhibition on TJ proteins expression. Conclusion: SAA can improve the Ber absorption in small intestine, through the PPs-associated immunity induced dynamic regulation on intestinal tissue permeability and TJ proteins expression. These findings might enlighten the research of traditional Chinese medicine decoction.
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In addition to defense response, phytohormone jasmonate participates in various plant growth and developmental processes. Nonetheless, its role in the seedling stage is not well defined. We recently report that jasmonate suppresses hypocotyl elongation and promotes cotyledon unfolding in etiolated Arabidopsis seedlings. The molecular basis underlying this phenotype is that jasmonate treatment reduces the biochemical activity of CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) to stabilize several COP1-targetted transcription factors for eliciting a proportion of light responsive transcriptome. We further reveal that jasmonate receptor CORONATINE INSENSITIVE 1 (COI1) and bHLH transcription factor MYC2 are required for the suppression of COP1 activity. Because elongated hypocotyl and closed cotyledons secure the success for seedling soil emergence, here we investigate seedling soil emergence under jasmonate treatment and find that jasmonate reduces the seedling emerging rates in the wild-type plants. Consistent with the largely insensitive to jasmonate in the suppression of skotomorphogenesis, the soil emerging rates in coi1 or myc2 mutants are almost not altered in the presence of jasmonate. Our data addendum describe that jasmonate-triggered inhibition of etiolation growth results in the defects in seedling soil emergence and suggest that defense-stimulated jasmonate biosynthesis might affect seedling germination in soil.
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Arabidopsis/fisiología , Ciclopentanos/farmacología , Oxilipinas/farmacología , Plantones/fisiología , Suelo , Arabidopsis/efectos de los fármacos , Plantones/efectos de los fármacosRESUMEN
Protein-protein interactions are fundamental mechanisms for relaying signal transduction in most cellular processes; therefore, identification of novel protein-protein interaction pairs and monitoring protein interaction dynamics are of particular interest for revealing how plants respond to environmental factors and/or developmental signals. A plethora of approaches have been developed to examine protein-protein interactions, either in vitro or in vivo. Among them, the recently established luciferase complementation imaging (LCI) assay is the simplest and fastest method for demonstrating in vivo protein-protein interactions. In this assay, protein A or protein B is fused with the amino-terminal or carboxyl-terminal half of luciferase, respectively. When protein A interacts with protein B, the two halves of luciferase will be reconstituted to form a functional and active luciferase enzyme. Luciferase activity can be recorded with a luminometer or CCD-camera. Compared with other approaches, the LCI assay shows protein-protein interactions both qualitatively and quantitatively. Agrobacterium infiltration in Nicotiana benthamiana leaves is a widely used system for transient protein expression. With the combination of LCI and transient expression, these approaches show that the physical interaction between COP1 and SPA1 was gradually reduced after jasmonate treatment.
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Luciferasas/metabolismo , Nicotiana/metabolismo , Proteínas de Plantas/metabolismo , Luciferasas/análisis , Hojas de la Planta/química , Hojas de la Planta/enzimología , Hojas de la Planta/metabolismo , Proteínas de Plantas/análisis , Dominios y Motivos de Interacción de Proteínas , Nicotiana/química , Nicotiana/enzimologíaRESUMEN
Derepression of transcription factors is the key mechanism for triggering plant jasmonate (JA) responses. Unlike regulating certain physiological functions for the majority of transcription factors in JA signaling, MYC2 and EIN3 control more diverse aspects. MYC2 predominantly participates in wounding response, metabolism, and root growth inhibition, while EIN3 (and its closest homolog EIL1) regulates defense gene expression and root hair development. Recently, it was reported that MYC2 and EIN3/EIL1 proteins mutually interact with each other and suppress their interaction partner's transcriptional activities. To understand their contributions in the modulation of transcriptomic network, we initially identified 1,495 differentially expressed jasmonate (JA)-responsive genes in wild-type Arabidopsis through RNA-seq analysis. Among them, 25% or 4.2% were independently regulated by EIN3/EIL1 or MYC2, respectively. Further analysis showed that EIN3/EIL1 and MYC2 interdependently regulate 16.3% of the JA-regulated transcriptome, including downregulation of three auxin-related genes, which might confer JA-inhibited root elongation. Lastly, we found that <30 genes were antagonistically regulated by MYC2 and EIN3/EIL1. We conclude that EIN3/EIL1 play a dominant role while MYC2 largely relies on EIN3/EIL1 for executing its transcriptional activity, either synergistically or antagonistically.
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ETHYLENE INSENSITIVE 2 (EIN2), an endoplasmic reticulum (ER) localized protein, plays a central role in relaying the ethylene signal from ER perception to the nucleus. Two recent reports reveal the novel role for EIN2 in translational control, providing another layer of regulation for ethylene signal transduction.