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Background: Long noncoding RNAs (lncRNAs), as emerging regulators of a wide variety of biological processes via diverse mechanisms, have been demonstrated to be of increasing importance in biology. Genome-wide association studies of tumor samples have identified several lncRNAs as either oncogenes or tumor suppressors in various types of cancers. In recent years, the importance of lncRNAs, especially in endometrioid cancer (EEC), has become increasingly well understood. The lncRNA Forkhead box P4 antisense RNA 1 (FOXP4-AS1) has been reported to fulfill roles in several types of cancers; however, the main biological function and associated underlying molecular mechanism of FOXP4-AS1 in EEC have yet to be fully elucidated. The present study therefore aimed to investigate how RNA FOXP4-AS1 may participate in the development and progression of endometrioid carcinoma tissues. Materials and Methods: In the present study, the expression level of FOXP4-AS1 was investigated in endometrioid carcinoma tissues and matching nearby normal endometrial tissues collected from patients receiving surgery at the hospital. A series of molecular biological assays were performed to investigate the effect of FOXP4-AS1 on cell proliferation, cell migration, and cell invasion. Results: An increased concentration of FOXP4-AS1 was identified in endometrioid carcinoma samples and cell lines compared with the corresponding controls, and this lncRNA was found to be positively correlated with advanced FIGO stages in patients with endometrial cancer. Furthermore, knocking down endogenous FOXP4-AS1 led to a significant reduction in the colony formation number and a significant inhibition of cell proliferation, cell migration, and cell invasion in endometrioid carcinoma cells. Moreover, dual-specificity phosphatase 5 (DUSP5), which is lowly expressed in endometrioid carcinoma tissues cells and negatively modulated by FOXP4-AS1, was identified as the downstream target molecule of FOXP4-AS1. Subsequently, the mechanistic experiments confirmed that, through binding to enhancer of zeste homolog 2 (EZH2; one of the catalytic subunits of polycomb repressive complex 2 [PRC2]), FOXP4-AS1 could epigenetically suppress the expression of DUSP5. Finally, the oncogenic function of the FOXP4-AS1/EZH2/DUSP5 axis in endometrioid carcinoma was confirmed via rescue assays. Conclusions: The findings of the present study have highlighted how FOXP4-AS1 fulfills an oncogenic role in endometrioid carcinoma, and targeting FOXP4-AS1 and its pathway may provide new biomarkers for patients with endometrioid carcinoma.
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Carcinoma Endometrioide , Fosfatasas de Especificidad Dual , Neoplasias Endometriales , ARN Largo no Codificante , Femenino , Humanos , Persona de Mediana Edad , Carcinoma Endometrioide/genética , Carcinoma Endometrioide/patología , Carcinoma Endometrioide/metabolismo , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Progresión de la Enfermedad , Fosfatasas de Especificidad Dual/genética , Fosfatasas de Especificidad Dual/metabolismo , Neoplasias Endometriales/genética , Neoplasias Endometriales/patología , Neoplasias Endometriales/metabolismo , Epigénesis Genética , Regulación Neoplásica de la Expresión Génica , Silenciador del Gen , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismoRESUMEN
BACKGROUND: Accumulating studies have reported indispensable functions of circular RNAs (circRNA) in tumor progression through regulation of gene expression. However, circRNA expression profiles and functions in human ovarian carcinoma (OC) are yet to be fully established. METHODS: In this research, deep sequencing of circRNAs from OC samples and paired adjacent normal tissues was performed to establish expression profiles and circ-PHC3 levels between the groups further compared using RT-qPCR. The effects of ectopic overexpression of miR-497-5p and SOX9 and siRNA-mediated knockdown of circ-PHC3 and an miR-497-5p inhibitor were explored to clarify the regulatory mechanisms underlying circ-PHC3 activity in OC proliferation and metastasis. Information from public databases and the luciferase reporter assay were further utilized to examine the potential correlations among circ-PHC3, miR-497-5p and SOX9. RESULTS: Our results showed significant upregulation of circ-PHC3 in both OC cell lines and tissues. In the luciferase reporter assay, downregulation of circ-PHC3 led to suppression of metastasis and proliferation, potentially through targeted effects on the miR-497-5p/SOX9 axis in OC. SOX9 overexpression or miR-497-5p suppression rescued OC cell proliferation and invasion following silencing of circ-PHC3. Moreover, SOX9 inhibition induced restoration of OC cell invasion and proliferation under conditions of overexpression of miR-497-5p. Thus, circ-PHC3 appears to exert effects on cancer stem cell differentiation through regulation of the miR-497-5p/SOX9 axis. CONCLUSION: Taken together, our findings suggest that circ-PHC3 enhances OC progression through functioning as an miR-497-5p sponge to promote SOX9 expression, supporting its potential as a promising candidate target for OC therapy.
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Carcinoma , MicroARNs , Neoplasias Ováricas , ARN Circular , Factor de Transcripción SOX9 , Femenino , Humanos , Carcinoma Epitelial de Ovario , Línea Celular Tumoral , Proliferación Celular/genética , MicroARNs/genética , Neoplasias Ováricas/genética , ARN Circular/genética , Factor de Transcripción SOX9/genéticaRESUMEN
The miR-141-3p has been reported to participate in regulating autophagy and tumor-stroma interactions in ovarian cancer (OC). We aim to investigate whether miR-141-3p accelerates the progression of OC and its effect on macrophage 2 polarization by targeting the Kelch-like ECH-associated protein1-Nuclear factor E2-related factor2 (Keap1-Nrf2) pathway. SKOV3 and A2780 cells were transfected with miR-141-3p inhibitor and negative control to confirm the regulation of miR-141-3p on OC development. Moreover, the growth of tumors in xenograft nude mice treated by cells transfected with miR-141-3p inhibitor was established to further testify the role of miR-141-3p in OC. The expression of miR-141-3p was higher in OC tissue compared with non-cancerous tissue. Downregulation of miR-141-3p inhibited the proliferation, migration, and invasion of ovarian cells. Furthermore, miR-141-3p inhibition also suppressed M2-like macrophage polarization and in vivo OC progression. Inhibition of miR-141-3p significantly enhanced the expression of Keap1, the target gene of miR-141-3p, and thus downregulated Nrf2, while activation of Nrf2 reversed the reduction in M2 polarization by miR-141-3p inhibitor. Collectively, miR-141-3p contributes to tumor progression, migration, and M2 polarization of OC by activating the Keap1-Nrf2 pathway. Inhibition of miR-141-3p attenuates the malignant biological behavior of ovarian cells by inactivating the Keap1-Nrf2 pathway.
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PTFE/Al reactive material is an energetic material that releases energy under impact conditions, resulting in a wide range of application prospects. In order to improve its damage ability-considering the higher heat of the reaction per unit mass when Ni2O3 is involved in the aluminothermic reaction-we designed and studied PTFE/Al/Ni2O3, a reaction material based on polytetrafluoroethylene (PTFE). We also designed two other kinds (PTFE/Al, PTFE/Al/CuO) for comparative study, with the mass fraction of the metal oxides added at 10%, 20%, and 30%, respectively. The quasi-static compression properties and impact initiation behavior of the material were investigated by a universal material testing machine and a drop hammer test. The reaction process of different materials under a high strain rate was recorded using a high-speed camera. The results show that with the increase in Ni2O3 content, the strength of the PTFE/Al/Ni2O3 reactive material shows an increasing trend followed by a decreasing trend. Among the three reactive materials, when the content of Al/Ni2O3 reaches 30 wt.%, the reaction duration is the longest (at 4 ms) and the reaction fireball is the largest. The addition of Ni2O3 is helpful to improve the reactivity and reaction duration of the PTEF/Al reactive material.
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Heat-induced gel-assisted desalination could efficiently and inexpensively remove salt from salted egg whites. However, it was at the expense of the excellent foaming properties of egg whites, caused by the denaturation and aggregation of proteins during heating treatment. Hence, in this current work, the enzymatic treatment was used to re-endow duck egg white nanogels (DEWN) with outstanding foaming properties. We found that low levels of hydrolysis (DH = 2.27 %) could dramatically improve the foaming capability (FC), reaching >200 %, which also enhanced the foaming stability (FS). As the hydrolysis time extended, the adsorption and diffusion rate of the supernatant on the interface increased and performed high elasticity. The dilatational rheology and Lissajous plots were explored to investigate the nonlinear dilatational rheological behaviors of the air/water interface stabilized by the hydrolysed samples. Finally, we evaluated the effect of pH on foaming properties and found that the FC could exceed 250 %, and the FS was close to 80 % at pH = 5. These encouraging results showed that simple enzymatic treatment could revive nanogels from their dissatisfied foaming properties. In this work, gel-assisted desalination combined with enzyme treatment significantly promotes the high-quality and high-value utilization of salted egg white.
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Patos , Clara de Huevo , Animales , Clara de Huevo/química , Hidrólisis , NanogelesRESUMEN
Cellulose acetate (CA) based films are widely used in liquid crystal displays due to their outstanding transparency, and a certain orientation birefringence of CA films is required when they are used as retardation films. The regulation of orientation birefringence is usually from the perspective of stretch-induced orientation, while the effects of crystallization behaviors of CA films remain obscure. In this study, the roles of crystallization and orientation on the orientation birefringence of CA films were elucidated. For cellulose diacetate films, the orientation birefringence is dominated by the orientation degree. In comparison, apart from the orientation degree, crystallinity is another key variable to regulate the orientation birefringence of cellulose triacetate and plasticized cellulose triacetate films, originating from the birefringence heterogeneity of the crystalline and amorphous phases. These results provide valuable guidelines for the production of CA-based optical films with excellent optical performance.
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Cristalización , Birrefringencia , Celulosa/análogos & derivadosRESUMEN
The cost-effective synthesis of flexible energy storage devices with high energy and power densities is a challenge in wearable electronics. Here, we report a facile, efficient, and scalable approach for preparing three-dimensional (3D) laser-induced graphene foam (Co3O4@LIG) embedded with porous Co3O4 nanocrystals using a CO2 infrared laser. The in situ formed Co3O4@LIG nanocomposites directly serve as active materials, current collectors, and the conductive substrate for micro-supercapacitors (MSCs). Benefiting from rational structural features, the MSC based on Co3O4@LIG nanocomposites (Co3O4@LIG-MSC) with an interdigitated electrode configuration exhibits excellent electrochemical performance, including a high specific capacitance (143.5 F g-1), excellent rate capability, high energy density (19.9 W h kg-1 at a power density of 0.5 W kg-1), and remarkable power density (15.0 W kg-1 at an energy density of 15.8 W h kg-1). Furthermore, the device possesses good stability under different bending diameters and cycling stability. Such a highly integrated flexible MSC with high energy and power densities made by a directly laser scribing strategy has some potential for the fabrication of wearable energy storage devices.
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KEY MESSAGE: Association and linkage mapping techniques were used to identify and verify single nucleotide polymorphisms (SNPs) associated with Sclerotinia sclerotiorum resistance. A novel resistant gene, GmGST , was cloned and shown to be involved in soybean resistance to SSR. Sclerotinia stem rot (SSR), caused by the fungus Sclerotinia sclerotiorum, is one of the most devastating diseases in soybean (Glycine max (Linn.) Merr.) However, the genetic architecture underlying soybean resistance to SSR is poorly understood, despite several mapping and gene mining studies. In the present study, the identification of quantitative trait loci (QTLs) involved in the resistance to S. sclerotiorum was conducted in two segregating populations: an association population that consisted of 261 diverse soybean germplasms, and the MH population, derived from a cross between a partially resistant cultivar (Maple arrow) and a susceptible cultivar (Hefeng25). Three and five genomic regions affecting resistance were detected by genome-wide association study to control the lesion length of stems (LLS) and the death rate of seedling (DRS), respectively. Four QTLs were detected to underlie LLS, and one QTL controlled DRS after SSR infection. A major locus on chromosome (Chr.) 13 (qDRS13-1), which affected both DRS and LLS, was detected in both the natural population and the MH population. GmGST, encoding a glutathione S-transferase, was cloned as a candidate gene in qDRS13-1. GmGST was upregulated by the induction of the partially resistant cultivar Maple arrow. Transgenic experiments showed that the overexpression of GmGST in soybean increased resistance to S. sclerotiorum and the content of soluble pigment in stems of soybean. The results increase our understanding of the genetic architecture of soybean resistance to SSR and provide a framework for the future marker-assisted breeding of resistant soybean cultivars.
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Ascomicetos/fisiología , Mapeo Cromosómico/métodos , Resistencia a la Enfermedad/inmunología , Glutatión Transferasa/metabolismo , Glycine max/genética , Enfermedades de las Plantas/inmunología , Proteínas de Plantas/metabolismo , Cromosomas de las Plantas/genética , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas , Genoma de Planta , Estudio de Asociación del Genoma Completo , Glutatión Transferasa/genética , Desequilibrio de Ligamiento , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Polimorfismo de Nucleótido Simple , Glycine max/crecimiento & desarrollo , Glycine max/microbiologíaRESUMEN
Strain-induced crystallization (SIC) in polybutadiene rubber (BR) was studied by in situ synchrotron radiation wide-angle X-ray diffraction (SR-WAXD) over a broad temperature range (-90 °C â 25 °C). Depending on the presence or absence of SIC and quiescent crystallization temperature, three temperature regions are divided. Detailed structural evolution is summarized in the strain-temperature space. Based on this micro-structural evolution information, the macroscopic mechanical response of BR, together with poly(isobutylene-isoprene) rubber (IIR) and natural rubber (NR), is reproduced based on Flory's and Plagge's theories. The origins of the mismatch of calculated and experimental stress-strain curves, especially in the large strain region, are discussed, and are mainly ascribed to the micro-macro connection approach and the network inhomogeneity.
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BACKGROUND: The hundred seed weight (HSW) is one of the yield components of soybean [Glycine max (L.) Merrill] and is especially critical for various soybean food types. In this study, a representative sample consisting of 185 accessions was selected from Northeast China and analysed in three tested environments to determine the quantitative trait nucleotide (QTN) of HSW through a genome-wide association study (GWAS). RESULT: A total of 24,180 single nucleotide polymorphisms (SNPs) with minor allele frequencies greater than 0.2 and missing data less than 3% were utilized to estimate linkage disequilibrium (LD) levels in the tested association panel. Thirty-four association signals were identified as associated with HSW via GWAS. Among them, nineteen QTNs were novel, and another fifteen QTNs were overlapped or located near the genomic regions of known HSW QTL. A total of 237 genes, derived from 31 QTNs and located near peak SNPs from the three tested environments in 2015 and 2016, were considered candidate genes, were related to plant growth regulation, hormone metabolism, cell, RNA, protein metabolism, development, starch accumulation, secondary metabolism, signalling, and the TCA cycle, some of which have been found to participate in the regulation of HSW. A total of 106 SNPs from 16 candidate genes were significantly associated with HSW in soybean. CONCLUSIONS: The identified loci with beneficial alleles and candidate genes might be valuable for the molecular network and MAS of HSW.
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Genes de Plantas/genética , Estudio de Asociación del Genoma Completo , Glycine max/crecimiento & desarrollo , Glycine max/genética , Semillas/crecimiento & desarrollo , Polimorfismo de Nucleótido SimpleRESUMEN
With the combination of a low-temperature extension rheometer and in situ synchrotron radiation wide-angle X-ray diffraction (SR-WAXD), the strain-induced crystallization (SIC) of poly(isobutylene-isoprene) rubber (IIR) was studied in the low-temperature region (-60 °C â 25 °C). The detailed structural evolution of IIR during the SIC is summarized in the strain-temperature space, where three distinct temperature zones are defined. The absence of the SIC in zone I (T > 0 °C) results in the poorest drawability of IIR among all measured temperatures. And with respect to the lowest temperature zone III (-60 °C < T < -50 °C), the SIC still occurs with low ultimate crystallinity (ca. 0.9%). More complicated structural evolution induced by the strain occurs in the intermediate-temperature zone II (-50 °C ≤ T ≤ 0 °C). The orientation ratio of the amorphous part Oa increases monotonically with the increment of the strain, but reaches a platform with Hencky strain ε > ca. 1.8. Meanwhile, the strain-induced crystal growth of IIR is evidenced by the dramatic increment of the lateral crystallite size of (110) and (113) planes. Moreover, the retraction experiment further reveals the network evolutions of IIR: suffering from low ultimate crystallinity (
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Non-alcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) is the most common chronic liver disease in the world. However, there are still no drugs for NAFLD/NASH in the market. Gastrodin (GAS) is a bioactive compound that is extracted from Gastrodia elata, which is used as an active compound on nervous system diseases. Recent reports showed that GAS and Gastrodia elata possess anti-oxidant activity and lipid regulating effects, which makes us curious to reveal the anti-NAFLD effect of GAS. A high cholesterol diet (HCD) was used to induce a NAFLD larval zebrafish model, and the lipid regulation and anti-oxidant effects were tested on the model. Furthermore, qRT-PCR studied the underlying mechanism of GAS. To conclude, this study revealed a lipid regulation and anti-oxidant insights of GAS on NAFLD larval zebrafish model and provided a potential therapeutic compound for NAFLD treatment.
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Antioxidantes/uso terapéutico , Alcoholes Bencílicos/uso terapéutico , Gastrodia/química , Glucósidos/uso terapéutico , Metabolismo de los Lípidos/efectos de los fármacos , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Animales , Antioxidantes/aislamiento & purificación , Alcoholes Bencílicos/aislamiento & purificación , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Glucósidos/aislamiento & purificación , Larva/efectos de los fármacos , Larva/genética , Larva/metabolismo , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patología , Pez Cebra/genética , Pez Cebra/metabolismoRESUMEN
Strain-induced crystallization (SIC) of natural rubber (NR) at descending temperatures as low as -60 °C is systematically investigated by in situ synchrotron radiation wide-angle X-ray diffraction (SR-WAXD) measurement. The detailed structural evolution of NR during SIC is studied in the strain-temperature space, where up to four regions are defined depending on the SR-WAXD results. In region I, the molecular chains begin to be oriented under tensile loading. The onset of crystallization happens in the very beginning of region II, and the NR crystal acts as a new physical cross-linking point to form a crystal network, namely the series model. The further increment of crystallinity (> ca. 8%) leads to the transition of the crystal network from the series model to the parallel model in region III. The crystal network is finally accomplished in region IV, where the crystallinity remains almost constant. Interestingly, regions III and IV exist only in the intermediate-temperature zone II (-40 °C to -10 °C), which are missing in zones I (-10 °C to 25 °C) and III (-60 °C to -40 °C). This suggests that sufficient crystallinity (χII-III > ca. 8%) is required to form the parallel model. The new crystal network provides a deep understanding of SIC of NR considering the microscopic features, i.e. oriented amorphous component, the onset of crystallization and crystallinity evolution and its correlation with the macroscopic stress-strain curve.
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The space-time properties of the expressions of sub-cycle and single-cycle focused optical pulses with radially polarized electromagnetic field based on the Sink-Source model are studied. The self-induced blue shift of the center frequency of spectrum in the center of the pulse field is found to have an important impact on the electrons acceleration. When the electrons approach to the center of pulse, the electrons will obtain a large kinetic energy gain in a short time. The effect of radiation-reaction force can't be ignored if the net kinetic energy gain of electrons is more than GeVs. The electrons will deviate from the original acceleration channel and the gain of kinetic energy that electrons may gain will be greatly reduced if the radiation-reaction effect is considered. In contrast to the few-cycle laser pulse accelerating electrons, the gain of kinetic energy obtained by electrons is a few times higher and the corresponding peak optical power is one order of magnitude lower in the case of the sub-cycle laser pulses accelerating electrons. The maximal kinetic energy gain of electrons is robust against the variation of the incident angles.
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The silage quality of forage soybean (FS) rich in protein with crop corn (CN) or sorghum (SG) rich in water soluble carbohydrate was investigated, and microbial community after ensiling was analyzed. Results showed that pH in mixed silages dropped to 3.5-3.8 lower than 100%FS silage (4.5). Microbial analysis indicated that mixed ensiling could influence the microbial community. Although Lactobacillus and Weissella were the dominant genera in all silage samples, Lactobacillus abundance in mixed silages (33-76%) was higher compared with 100%FS silage (27%). In conclusion, FS ensiled with CN or SG could be an alternative approach to improve FS silage quality.
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Fermentación , Ensilaje , Sorghum , Glycine max , Zea maysRESUMEN
A series of [1,2,4]triazolo[3,4-b][1,3,4]thiadiazole derivatives were designed, synthesized and evaluated for their biological activity. Most of these compounds showed potent activities against c-Met kinase and cell growth inhibition. The most promising compound, 7d, has the IC50 values of 2.02 and 88â¯nM to inhibit c-Met kinase activity and cell growth in the MKN45â¯cell line, respectively. In addition, 7d is highly selective to c-Met and exhibits over 2500-fold selective inhibition to 16 tyrosine kinases evaluated.
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Antineoplásicos/farmacología , Descubrimiento de Drogas , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-met/antagonistas & inhibidores , Tiadiazoles/farmacología , Triazoles/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Simulación del Acoplamiento Molecular , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Proteínas Proto-Oncogénicas c-met/metabolismo , Relación Estructura-Actividad , Tiadiazoles/síntesis química , Tiadiazoles/química , Triazoles/síntesis química , Triazoles/químicaRESUMEN
c-Met was emerging as an attractive target for cancer-targeted therapy because deregulation of c-Met has been observed in multiple tumor types. A series of 4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine derivatives were designed, synthesized and evaluated for their enzymatic inhibitory activity against c-Met kinase and cellular potency against MKN45, EBC-1 and PC-3 cell lines. Nine of them showed better activity than lead compound 1 which was found via computer-aided drug design. Among them, compound 8c showed inhibitory activity of 68 nM against c-Met and low micromole cellular potency against MKN45 and EBC-1 cell lines. Moreover, 8c demonstrated more than 50-fold selectivity against other tyrosine kinases tested. The result of western blot indicated that compound 8c was capable of inhibiting the phosphorylation of c-Met kinase in MKN45 cell line in a dose-dependent manner.
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Diseño de Fármacos , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-met/antagonistas & inhibidores , Pirazoles/farmacología , Piridinas/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Proteínas Proto-Oncogénicas c-met/metabolismo , Pirazoles/síntesis química , Pirazoles/química , Piridinas/síntesis química , Piridinas/química , Relación Estructura-ActividadRESUMEN
Enantioselective total syntheses of (+)-fawcettidine and (+)-lycoposerramine Q as well as the first total synthesis of (-)-lycopladine D from a common intermediate have been accomplished by a divergent path. The common intermediate was derived from a Hajos-Parrish-like diketone by a stereoselective Birch reduction and a Suzuki coupling. The synthesis of (-)-lycopladine D featured an allylic oxidation and a biomimetic aminoketalization while the route to (+)-fawcettidine and (+)-lycoposerramine Q highlighted an oxidative rearrangement.
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Alcaloides/síntesis química , Sesquiterpenos/síntesis química , Alcaloides/química , Compuestos Heterocíclicos con 3 Anillos , Estructura Molecular , Oxidación-Reducción , Sesquiterpenos/química , EstereoisomerismoRESUMEN
BACKGROUND: Soybean [Glycine max (L.) Merr.] is one of the most important crops for oil and protein resource. Improvement of stress tolerance will be beneficial for soybean seed production. PRINCIPAL FINDINGS: Six GmPHD genes encoding Alfin1-type PHD finger protein were identified and their expressions differentially responded to drought, salt, cold and ABA treatments. The six GmPHDs were nuclear proteins and showed ability to bind the cis-element "GTGGAG". The N-terminal domain of GmPHD played a major role in DNA binding. Using a protoplast assay system, we find that GmPHD1 to GmPHD5 had transcriptional suppression activity whereas GmPHD6 did not have. In yeast assay, the GmPHD6 can form homodimer and heterodimer with the other GmPHDs except GmPHD2. The N-terminal plus the variable regions but not the PHD-finger is required for the dimerization. Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants. This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes. SIGNIFICANCE: These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.
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Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Glycine max/genética , Proteínas de Soja/genética , Factores de Transcripción/genética , Secuencia de Aminoácidos , Clonación Molecular , Dimerización , Perfilación de la Expresión Génica , Datos de Secuencia Molecular , Plantas Modificadas Genéticamente , Unión Proteica , Estructura Terciaria de Proteína , Protoplastos/metabolismo , Homología de Secuencia de Aminoácido , Estrés FisiológicoRESUMEN
A full-length gene GmSAMDC1, encoding the S-adenosylmethionine decarboxylase (SAMDC), a key enzyme involved in polyamine biosynthesis, was identified from soybean expressed sequence tags and was characterized. GmSAMDC1 encoded a peptide of 355 amino acids. When compared with other plant SAMDCs, the GmSAMDC1 protein had several highly conserved regions including a putative pro-enzyme cleavage site and a PEST sequence. The 5' leader sequence of the the GmSAMDC1 mRNA contained two additional open reading frames (ORFs), which may regulate the translational process. The genomic sequence of the GmSAMDC1 gene contained three introns in the 5' leader sequence, but no intron in the 3'-UTR or the main pro-enzyme ORF. A simple sequence repeat (SSR) was found in intron 2, and the GmSAMDC1 gene was mapped to linkage group D1 using this SSR. The genomic organization of the GmSAMDC1 gene in the subgenus Glycine and the subgenus Soja was found to be different by Southern-blot and PCR analysis. A pseudogene, GmSAMDC2, was also identified. This gene contained no intron and lost its two uORFs. Northern-blot analysis showed that the GmSAMDC1 gene expression was induced by salt, drought and cold, but not induced by wounding; suggesting that the gene was implicated in response to multiple-stress conditions.