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Histone modifications are critical epigenetic indicators of chromatin state associated with gene expression. Although the reprogramming patterns of H3K4me3 and H3K27me3 have been elucidated in mouse and human preimplantation embryos, the relationship between these marks and zygotic genome activation (ZGA) remains poorly understood. By ultra-low-input native chromatin immunoprecipitation and sequencing, we profiled global H3K4me3 and H3K27me3 in porcine oocytes and in vitro fertilized (IVF) embryos. We found that promoters of ZGA genes occupied sharp H3K4me3 peaks in oocytes, and these peaks became broader after fertilization, and reshaped into sharp again during ZGA. By simultaneous depletion of H3K4me3 demethylase KDM5B and KDM5C, we determined that broad H3K4me3 domain maintenance impaired ZGA gene expression, suggesting its function to prevent premature ZGA entry. By contrast, broad H3K27me3 domains underwent global removal upon fertilization, followed by a re-establishment for H3K4me3/H3K27me3 bivalency in morulae. We also found that bivalent marks were deposited at promoters of ZGA genes, and inhibiting this deposition was correlated with the activation of ZGA genes. It suggests that promoter bivalency contributes to ZGA exit in porcine embryos. Moreover, we demonstrated that aberrant reprogramming of H3K4me3 and H3K27me3 triggered ZGA dysregulation in somatic cell nuclear transfer (SCNT) embryos, whereas H3K27me3-mediated imprinting did not exist in porcine IVF and SCNT embryos. Our findings highlight two previously unknown epigenetic reprogramming modes coordinated with ZGA in porcine preimplantation embryos. Finally, the similarities observed between porcine and human histone modification dynamics suggest that the porcine embryo may also be a useful model for human embryo research.
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The regulatory network between signaling pathways and transcription factors (TFs) is crucial for the maintenance of pluripotent stem cells. However, little is known about how the key TF OCT4 coordinates signaling pathways to regulate self-renewal and lineage differentiation of porcine pluripotent stem cells (pPSCs). Here, we explored the function of OCT4 in pPSCs by transcriptome and chromatin accessibility analysis. The TFs motif enrichment analysis revealed that, following OCT4 knockdown, the regions of increased chromatin accessibility were enriched with EOMES, GATA6, and FOXA1, indicating that pPSCs differentiated toward the mesoendoderm (ME) lineage. Besides, pPSCs rapidly differentiated into ME when the WNT/ß-catenin inhibitor XAV939 was removed. However, the ME differentiation of pPSCs caused by OCT4 knockdown did not rely on the activation of WNT/ß-catenin signaling because the target gene of WNT/ß-catenin signaling, AXIN2 was not upregulated after OCT4 knockdown, despite significant upregulation of WLS and some WNT ligands. Importantly, OCT4 is directly bound to the promoter and enhancers of EOMES and repressed its transcription. Overexpression of EOMES was sufficient to induce ME differentiation in the presence of XAV939. These results demonstrate that OCT4 can regulate WNT/ß-catenin signaling and prevent ME differentiation of pPSCs by repressing EOMES transcription.
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Diferenciação Celular , Células-Tronco Pluripotentes , Via de Sinalização Wnt , Animais , beta Catenina/genética , beta Catenina/metabolismo , Diferenciação Celular/genética , Cromatina/metabolismo , Células-Tronco Pluripotentes/citologia , Células-Tronco Pluripotentes/metabolismo , Suínos , Via de Sinalização Wnt/genética , Proteínas com Domínio T/metabolismo , Fator 3 de Transcrição de Octâmero/metabolismo , Linhagem CelularRESUMO
In brief: Normal gene expression during early embryonic development and in the placenta is crucial for a successful pregnancy. Nicotine can disrupt normal gene expression during development, leading to abnormal embryonic and placental development. Abstract: Nicotine is a common indoor air pollutant that is present in cigarette fumes. Due to its lipophilic nature, nicotine can rapidly transport through membrane barriers and spread throughout the body, which can lead to the development of diseases. However, the impact of nicotine exposure during early embryonic development on subsequent development remains elusive. In this study, we found that nicotine significantly elevated reactive oxygen species, DNA damage and cell apoptosis levels with the decrease of blastocyst formation during early embryonic development. More importantly, nicotine exposure during early embryonic development increased placental weight and disrupted placental structure. In molecular level, we also observed that nicotine exposure could specifically cause the hypermethylation of Phlda2 promoter (a maternally expressed imprinted gene associated with placental development) and reduce the mRNA expression of Phlda2. By RNA sequencing analysis, we demonstrated that nicotine exposure affected the gene expression and excessive activation of the Notch signaling pathway thereby affecting placental development. Blocking the Notch signaling pathway by DAPT treatment could recover abnormal placental weight and structure induced by nicotine exposure. Taken together, this study indicates that nicotine causes the declining quality of early embryos and leads to placental abnormalities related to over-activation of the Notch signaling pathway.
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Placenta , Placentação , Gravidez , Feminino , Humanos , Placenta/metabolismo , Nicotina/toxicidade , Nicotina/metabolismo , Proteínas Nucleares/metabolismo , Transdução de SinaisRESUMO
Glioma is the most common brain cancer with a poor prognosis, and its underlying molecular mechanisms still needs to be further explored. In the current study, we discovered that an antisense lncRNA, CACNA1C-AS2, suppressed growth, migration and invasion of glioma cells, suggesting that CACNA1C-AS2 functions as a tumor suppressor. Furthermore, we found that CACNA1C-AS2 negatively regulated Fbxo45 protein expression in glioma cells. Impressively, extensive experimental results revealed that Fbxo45 accelerated growth, migration and invasion of glioma cells. Clinically, increased Fbxo45 expression was observed in 75 human glioma tissue samples. Moreover, in vivo experiments also demonstrated that Fbxo45 overexpression enhanced tumor growth in mice. Especially, we further identified that Fbxo45 activated mTORC1 rather than mTORC2 through PI3K/AKT signaling to promote cell growth and motility in glioma cells. Rescue experiments also exhibited that CACNA1C-AS2 inhibited cell growth and motility partly through down-regulating Fbxo45 expression in glioma. Our results provide the novel insights into the critical role of CACNA1C-AS2/Fbxo45/mTOR axis involved in regulating glioma tumorigenesis and progression, and further indicate that CACNA1C-AS2 and Fbxo45 may be the potential biomarkers and therapeutic targets for glioma.
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Proteínas F-Box , Glioma , MicroRNAs , Humanos , Camundongos , Animais , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , MicroRNAs/genética , Linhagem Celular Tumoral , Apoptose/genética , Glioma/metabolismo , Proliferação de Células/genética , Movimento Celular/genética , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo , Canais de Cálcio Tipo L , Proteínas F-Box/genéticaRESUMO
In the shallow eutrophic lakes in cold, arid regions, the phytoplankton functional groups and the factors that drive their spatiotemporal variabilities remain unclear. Samples were collected from Lake Ulansuhai in April, August, and October 2017 (wet season) and January 2018 (dry season). Based on the functional group classification method, 23 phytoplankton functional groups with 5 major ones were identified. During the wet season, high amounts of nutrients, elevated temperatures, and heavy rainfall produced spatiotemporal variabilities in phytoplankton communities, whereas during the dry season, the frozen period was the critical factor that determined the spatiotemporal variabilities in the phytoplankton communities. Through redundancy analyses, total nitrogen and total phosphorus concentrations were observed to directly affect the phytoplankton growth; algal growth affected the chemical oxygen demand, and pH and environmental factors interacted with the phytoplankton growth. These results highlight the complex feedbacks of shallow eutrophic lake ecosystems in arid regions. Group TC (represented by Lyngbya) was correlated with Huangtai algae. In August, a Huangtai algal bloom resulted in a relatively stable water column, which was conducive to group TC growth. Therefore, the presence of certain phytoplankton functional groups can indicate the current lake conditions by identifying the coverage of Huangtai algae, which provides a scientific basis for an early warning of a potential algal bloom.
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Monitoramento Ambiental , Lagos , Fitoplâncton , China , Ecossistema , Eutrofização , FósforoRESUMO
Herein, through a dual-ligand strategy, we report eight isorecticular lanthanide(III) furan-2,5-dicarboxylic acid metal-organic frameworks (Ln-MOFs) with the general formula {[Ln(2,5-FDA)0.5(Glu)(H2O)2]· xH2O} n [Ln = Sm (1), Eu (2), Gd (3), Tb (4), Dy (5), Ho (6), Er (7), and Yb (8); 2,5-FDA2- = furan-2,5-dicarboxylate and Glu2- = glutarate; x = 0.5 for 1, 2, and 4 and x = 0 for 3 and 5-8], synthesized under solvothermal conditions by using an N, N'-dimethylformamide/H2O mixed solvent system. Crystallographic data reveal that all eight Ln-MOFs 1-8 crystallize in the orthorhombic Pnma space group. All of the MOFs are isostructural as well as isomorphous with distorted monocapped square-antiprismatic geometry around the Ln1 metal center. In Ln-MOFs 1-8, the 2,5-FDA2- and Glu2- ligands exhibit µ2-κ4,η1:η1:η1:η1 and µ3-κ5,η2:η1:η1:η1 coordination modes, respectively. Topologically, assembled Ln-MOFs 1-8 consist of the 2D cem topological type. The designed Ln-MOFs 1-8 are further explored for structure-corroborated density functional theory study. Meanwhile, room temperature photoluminescence properties of Ln-MOFs 2 and 4 and magnetic properties of Ln-MOFs 3 and 5 have been explored in detail. A highly intense, ligand-sensitized, Ln3+ f-f photoluminescence emission is exhibited by Ln-MOFs 2 [Eu3+ (red emission)] and 4 [Tb3+ (green emission)]. Magnetic studies suggest weak antiferro- and ferromagnetic interactions between adjacent GdIII ions in Ln-MOF 3, thereby displaying a large magnetocaloric effect. The magnetic data measured at T = 2 K and Δ H = 30 kOe depict that the -Δ Sm value per unit mass reaches 32.1 J kg-1 K-1, which is larger than most of the GdIII-based complexes reported. The alternating-current susceptibility measurements on Ln-MOF 5 revealed that out-of-phase signals are frequency- and temperature-dependent under both 0 and 2 kOe direct-current fields, thereby suggesting a typical slow magnetic relaxation behavior with two relaxation processes. This is further supported by the Cole-Cole plots at 2.4-6 K.
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The common approach in fluorescence molecular tomography (FMT) assumes homogeneous distributions of the optical properties and normally results in reconstructions of low sensitivity. A natural enhancement is to incorporate diffuse optical tomography (DOT) to FMT. However, the traditional voxel-based DOT has been a severely ill-posed inverse problem and cannot retrieve the optical property distributions accurately. We present a structural-prior-based DOT method to effectively acquire the heterogeneous optical background with the aid of some imperfect structural priors from x-ray computed tomography and/or magnetic resonance imaging anatomical imaging modalities, and quantitatively compare its hard- and soft-prior schemes for achieving an improved recovery of the fluorescence distribution. Numerical simulations are conducted on a region-labeled three-dimensional (3D) digital mouse model to investigate the performance of this method. Physical experiments on a cylindrical phantom are also conducted to assess this methodology. Our simulated and experimental reconstruction results indicate that the structural-prior-based DOT guided FMT approach can significantly improve the sensitivity of FMT reconstruction, as well as its imaging resolution and quantitative accuracy.
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Interpretação de Imagem Assistida por Computador/métodos , Fígado/citologia , Fígado/metabolismo , Microscopia de Fluorescência/métodos , Imagem Molecular/métodos , Tomografia Óptica/métodos , Tomografia Computadorizada por Raios X/métodos , Algoritmos , Animais , Camundongos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The effective proliferation and differentiation of trophoblast stem cells (TSCs) is indispensable for the development of the placenta, which is the key to maintaining normal fetal growth during pregnancy. Kruppel-like factor 5 (Klf5) is implicated in the activation of pluripotency gene expression in embryonic stem cells (ESCs), yet its function in TSCs is poorly understood. Here, we showed that Klf5 knockdown resulted in the downregulation of core TSC-specific genes, consequently causing rapid differentiation of TSCs. Consistently, Klf5-depleted embryos lost the ability to establish TSCs in vitro. At the molecular level, Klf5 preferentially occupied the proximal promoter regions and maintained an open chromatin architecture of key TSC-specific genes. Deprivation of Klf5 impaired the enrichment of p300, a major histone acetyl transferase of H3 lysine 27 acetylation (H3K27ac), and further reduced the occupancy of H3K27ac at promoter regions, leading to decreased transcriptional activity of TSC pluripotency genes. Thus, our findings highlight a novel mechanism of Klf5 in regulating the self-renewal and differentiation of TSCs and provide a reference for understanding placental development and improving pregnancy rates.
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Placenta , Fatores de Transcrição , Feminino , Gravidez , Humanos , Placenta/metabolismo , Fatores de Transcrição/metabolismo , Trofoblastos/metabolismo , Diferenciação Celular/genética , Células-Tronco Embrionárias/metabolismoRESUMO
As global anthropogenic nitrogen inputs continue to rise, nitrite-dependent anaerobic methane oxidation (N-DAMO) plays an increasingly significant role in CH4 consumption in lake sediments. However, there is a dearth of knowledge regarding the effects of anthropogenic activities on N-DAMO bacteria in lakes in the cold and arid regions. Sediment samples were collected from five sampling areas in Lake Ulansuhai at varying depth ranges (0-20, 20-40, and 40-60 cm). The ecological characterization and niche differentiation of N-DAMO bacteria were investigated using bioinformatics and molecular biology techniques. Quantitative PCR confirmed the presence of N-DAMO bacteria in Lake Ulansuhai sediments, with 16S rRNA gene abundances ranging from 1.72 × 104 to 5.75 × 105 copies·g-1 dry sediment. The highest abundance was observed at the farmland drainage outlet with high available phosphorus (AP). Anthropogenic disturbances led to a significant increase in the abundance of N-DAMO bacteria, though their diversity remained unaffected. The heterogeneous community of N-DAMO bacteria was affected by interactions among various environmental characteristics, with AP and oxidation-reduction potential identified as the key drivers in this study. The Mantel test indicated that the N-DAMO bacterial abundance was more readily influenced by the presence of the denitrification genes (nirS and nirK). Network analysis revealed that the community structure of N-DAMO bacteria generated numerous links (especially positive links) with microbial taxa involved in carbon and nitrogen cycles, such as methanogens and nitrifying bacteria. In summary, N-DAMO bacteria exhibited sensitivity to both environmental and microbial factors under various human disturbances. This study provides valuable insights into the distribution patterns of N-DAMO bacteria and their roles in nitrogen and carbon cycling within lake ecosystems.
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Microbiota , Nitritos , Humanos , Lagos/microbiologia , Anaerobiose , Metano , RNA Ribossômico 16S/genética , Bactérias/genética , Methanobacteriaceae , Bactérias Anaeróbias/genética , Oxirredução , Nitrogênio , Carbono , DesnitrificaçãoRESUMO
Chromatin accessibility remodeling driven by pioneer factors is critical for the development of early embryos. Current studies have illustrated several pioneer factors as being important for agricultural animals, but what are the pioneer factors and how the pioneer factors remodel the chromatin accessibility in porcine early embryos is not clear. By employing low-input DNase-seq (liDNase-seq), we profiled the landscapes of chromatin accessibility in porcine early embryos and uncovered a unique chromatin accessibility reprogramming pattern during porcine preimplantation development. Our data revealed that KLF4 played critical roles in remodeling chromatin accessibility in porcine early embryos. Knocking down of KLF4 led to the reduction of chromatin accessibility in early embryos, whereas KLF4 overexpression promoted the chromatin openness in porcine blastocysts. Furthermore, KLF4 deficiency resulted in mitochondrial dysfunction and developmental failure of porcine embryos. In addition, we found that overexpression of KLF4 in blastocysts promoted lipid droplet accumulation, whereas knockdown of KLF4 disrupted this process. Taken together, our study revealed the chromatin accessibility dynamics and identified KLF4 as a key regulator in chromatin accessibility and cellular metabolism during porcine preimplantation embryo development.
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Cromatina , Desenvolvimento Embrionário , Suínos , Animais , Desenvolvimento Embrionário/genética , Cromatina/genética , Cromatina/metabolismo , Blastocisto/metabolismo , CromossomosRESUMO
Emerging evidence highlights the regulatory role of paired-like (PRD-like) homeobox transcription factors (TFs) in embryonic genome activation (EGA). However, the majority of PRD-like genes are lost in rodents, thus prompting an investigation into PRD-like TFs in other mammals. Here, we showed that PRD-like TFs were transiently expressed during EGA in human, monkey, and porcine fertilized embryos, yet they exhibited inadequate expression in their cloned embryos. This study, using pig as the research model, identified LEUTX as a key PRD-like activator of porcine EGA through genomic profiling and found that LEUTX overexpression restored EGA failure and improved preimplantation development and cloning efficiency in porcine cloned embryos. Mechanistically, LEUTX opened EGA-related genomic regions and established histone acetylation via recruiting acetyltransferases p300 and KAT2A. These findings reveal the regulatory mechanism of LEUTX to govern EGA in pigs, which may provide valuable insights into the study of early embryo development for other non-rodent mammals.
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Genoma , Técnicas de Transferência Nuclear , Animais , Suínos , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/metabolismo , Proteínas de Homeodomínio/genética , Desenvolvimento Embrionário/genética , Embrião de Mamíferos/metabolismo , Humanos , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Acetilação , Clonagem de Organismos/métodos , Histonas/metabolismo , Blastocisto/metabolismoRESUMO
BACKGROUND: Immune-related initiation, progress, metastasis and sensitivity to treatment associated with poor prognosis of patients with colorectal cancer (CRC). The role of Nucleolar protein interacting with the FHA domain of MKI67 (NIFK) in CRC remained to be investigated. We explore whether NIFK correlates with tumor immune infiltration and plays an important role in CRC patient prognosis. METHODS: The data of samples involved in our study was obtained from TCGA and GEO and samples for protein expression detection and clinical information analysis were obtained from our hospital. NIFK expression, association with patient prognosis, correlation with infiltration of immune cells and its correlated genes involved in signaling pathways were analyzed using bioinformatics method along with experimental validation and clinical correlation analysis. RESULTS: Results indicated that the expression of NIFK in tumor tissues was significantly increased compared with normal samples. colon and rectal cancer patients with high NIFK expression have poor survival compared with those with low NIFK expression. Results of cell experiments indicated that NIFK is positively correlated with cell proliferation and migration in CRC. NIFK negatively correlated with T cell CD8+, Tregs, Neutrophil and macrophage significantly. DARS and NKRF were positively correlated with NIFK and DARS correlated with CD8 + T cell, CD4 + T cell, macrophage and Neutrophil, NKRF correlated with CD8 + T cell, CD4 + T cell and macrophage in colon and rectal cancer. NIFK along with its correlated genes as DARS and NKRF were involved in Wnt, PI3K-Akt, NF-κB signaling and Intestinal immune network for lgA production. CONCLUSIONS: Our results suggested that NIFK might be a biomarker associated with poor prognosis of CRC patients, and it would be a potential target for CRC therapy.
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Neoplasias Colorretais , Neoplasias Retais , Humanos , Neoplasias Colorretais/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Prognóstico , Transdução de SinaisRESUMO
Arsenite is commonly used as an insecticide, antiseptic and herbicide. It can enter the food chain via through soil contamination, and harm human health, including the reproductive systems. Early embryos, as the initial stage of mammalian life, are very sensitive to the environmental toxins and pollutants. However, whether and how arsenite disturbs the early embryo development remains unclear. Our study used mouse early embryos as a model and revealed that arsenite exposure did not cause reactive oxygen species production, DNA damage or apoptosis. However, arsenite exposure arrested embryonic development at the 2-cell stage by altering gene expression patterns. The transcriptional profile in the disrupted embryos showed abnormal maternal-to-zygote transition (MZT). More importantly, arsenite exposure attenuated H3K27ac modification enrichment at the promoter region of Brg1, a key gene for MZT, which inhibited its transcription, and further affected MZT and early embryonic development. In conclusion our study highlights arsenite exposure affects MZT by reducing the enrichment of H3K27ac on the embryonic genome, and ultimately induces early embryonic development arrest at the 2-cell stage.
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Arsenitos , Zigoto , Gravidez , Feminino , Humanos , Animais , Camundongos , Zigoto/metabolismo , Arsenitos/toxicidade , Arsenitos/metabolismo , Desenvolvimento Embrionário/genética , Mamíferos/genética , Mamíferos/metabolismo , Regulação da Expressão Gênica no DesenvolvimentoRESUMO
Macroautophagy/autophagy is a cellular and energy homeostatic mechanism that contributes to maintain the number of primordial follicles, germ cell survival, and anti-ovarian aging. However, it remains unknown whether autophagy in granulosa cells affects oocyte maturation. Here, we show a clear tendency of reduced autophagy level in human granulosa cells from women of advanced maternal age, implying a potential negative correlation between autophagy levels and oocyte quality. We therefore established a co-culture system and show that either pharmacological inhibition or genetic ablation of autophagy in granulosa cells negatively affect oocyte quality and fertilization ability. Moreover, our metabolomics analysis indicates that the adverse impact of autophagy impairment on oocyte quality is mediated by downregulated citrate levels, while exogenous supplementation of citrate can significantly restore the oocyte maturation. Mechanistically, we found that ACLY (ATP citrate lyase), which is a crucial enzyme catalyzing the cleavage of citrate, was preferentially associated with K63-linked ubiquitin chains and recognized by the autophagy receptor protein SQSTM1/p62 for selective autophagic degradation. In human follicles, the autophagy level in granulosa cells was downregulated with maternal aging, accompanied by decreased citrate in the follicular fluid, implying a potential correlation between citrate metabolism and oocyte quality. We also show that elevated citrate levels in porcine follicular fluid promote oocyte maturation. Collectively, our data reveal that autophagy in granulosa cells is a beneficial mechanism to maintain a certain degree of citrate by selectively targeting ACLY during oocyte maturation.Abbreviations: 3-MA: 3-methyladenine; ACLY: ATP citrate lyase; AMA: advanced maternal age; CG: cortical granule; CHX: cycloheximide; CQ: chloroquine; CS: citrate synthase; COCs: cumulus-oocyte-complexes; GCM: granulosa cell monolayer; GV: germinal vesicle; MII: metaphase II stage of meiosis; PB1: first polar body; ROS: reactive oxygen species; shRNA: small hairpin RNA; SQSTM1/p62: sequestosome 1; TCA: tricarboxylic acid; TOMM20/TOM20: translocase of outer mitochondrial membrane 20; UBA: ubiquitin-associated domain; Ub: ubiquitin; WT: wild-type.
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ATP Citrato (pro-S)-Liase , Macroautofagia , Feminino , Humanos , Animais , Suínos , Proteína Sequestossoma-1/metabolismo , ATP Citrato (pro-S)-Liase/metabolismo , Ácido Cítrico/metabolismo , Autofagia , Oócitos/metabolismo , Citratos/metabolismo , Aciltransferases/metabolismo , Ubiquitina/metabolismo , HomeostaseRESUMO
Extensive efforts have been made to integrate diffuse optical tomography (DOT) with other imaging modalities, such as magnetic-resonance imaging and x-ray computerized tomography, for its performance improvement. However, the experimental apparatus is in general intricate and costly due to adoption of the physically distinct radiation regimes. In this study, a time-domain fluorescence-guided DOT methodology that incorporates a priori localization information provided by diffuse fluorescence tomography (DFT) is investigated in an attempt to optimize recovery of the optical property distributions. The methodology is based on a specifically designed multichannel time-correlated single-photon-counting DOT/DFT system as well as a featured-data image reconstruction scheme that is developed within the framework of the generalized pulse spectrum technique and employs the third-order simplified harmonics approximation to the radiative transfer equation as the forward model. We have validated the methodology using phantom experiments and demonstrated that, with the guidance of fluorescence a priori, the quantitativeness and spatial resolution of the recovered optical target can be considerably improved in terms of the absorption and scattering images.
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Aumento da Imagem/instrumentação , Microscopia de Fluorescência/instrumentação , Fotometria/instrumentação , Tomografia Óptica/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Imagens de FantasmasRESUMO
OBJECTIVE: To investigate the structure of ammonia-oxidation microbial communities in the wetlands to dry-up process at 99 degraded lakes of the Huitengxile grassland in the Inner Mongolia Plateau. METHODS: The microbial quantity of ammonia-oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) were examined by most probable number-polymerase chain reaction (MPN-PCR). The clone libraries of amoA were constructed and phylogenetics were analyzed. With analysis of the soil properties, we evaluated the effects of wetlands degradation on ammonia-oxidation microbes communities. RESULTS: In 75% of the samples, the quantity of AOB communities was higher than that of AOA; moreover, quantity of bacterial were up to 18.1-fold more abundant than Archaea's. The AOB microbial quantity was strongly correlated with NH4+-N content in the soil. Phylogenetic analyses of the amoA gene fragments showed that most AOB sequences from degraded wetlands were affiliated with Nitrosomonas-like species and a few close to Nitrosospira. All AOA sequences belonged to the kingdom Crenarchaeote. CONCLUSION: Experimental results showed that quantity of ammonia-oxidation microbes increased but community diversity declined during wetlands degradation , and oxidation conditions and ammonium concentration in the soil might play important roles in the community structure of both the AOA and AOB.
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Amônia/metabolismo , Archaea/isolamento & purificação , Bactérias/isolamento & purificação , Microbiologia do Solo , Lagos , Oxirredução , FilogeniaRESUMO
A new bacterial strain, Acinetobacter calcoaceticus TY1, was identified in activated sludge. This strain efficiently metabolized nitrogen from ammonium at low temperatures, utilizing NH4+-N, NO3--N, and NO2--N as nitrogen sources. Of these, NH4+-N was superior in terms of both assimilation and heterotrophic nitrification at 8 °C. The nitrogen metabolism-associated genes amoA, nirK, and nosZ were identified in TY1. Optimal requirements for growth and nitrogen removal were pH 7, shaking speed of 90 rpm, a C/N ratio of 10, and sodium citrate for the carbon supply. The ability to denitrify at low temperature suggests TY1's potential for wastewater management.
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Acinetobacter calcoaceticus , Compostos de Amônio , Acinetobacter calcoaceticus/genética , Acinetobacter calcoaceticus/metabolismo , Aerobiose , Bactérias/metabolismo , Desnitrificação , Processos Heterotróficos , Nitrificação , Nitritos/metabolismo , Nitrogênio/metabolismo , TemperaturaRESUMO
We report herein a palladium-catalyzed domino cyclization/carbonylation to access ester-functionalized azaindolines, applying formates as a convenient carbonyl source. All four azaindoline isomers were constructed, exhibiting good functional group compatibility. On this basis, modifying the starting tether on the aminopyridine led to furoazaindolines via an intramolecular reductive cyclization after the palladium-catalyzed process.
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Esophageal squamous cell carcinoma (ESCC) is one of the most common and deadly cancers. Fbxo45, a substrate recognition subunit of E3 ligase, is critically involved in tumorigenesis and tumor progression. However, the function of Fbxo45 and the underlying mechanisms have not been elucidated in ESCC. We used cellular and molecular methods to explore the molecular basis of Fbxo45-mediated ESCC development. We found that ectopic overexpression of Fbxo45 promoted the growth of Kyse-150, Kyse30 and ECA-109 cells and inhibited the apoptosis. Moreover, overexpression of Fbxo45 promoted the migration and invasion of ESCC cells. Consistently, knockdown of Fbxo45 exhibited the opposite effects on ESCC cells. Mechanistically, we observed that Fbxo45 binds to GGNBP2 via its SPRY domain and targets GGNBP2 for ubiquitination and degradation. GGNBP2 overexpression exhibited anticancer activity in ESCC cells. Furthermore, Fbxo45 exerted its functions by regulating GGNBP2 stability in ESCC cells. Notably, overexpression of Fbxo45 facilitated tumor growth in mice. Strikingly, Fbxo45 was highly expressed in ESCC tissues, and GGNBP2 had a lower expression in ESCC specimens. High expression of Fbxo45 and low expression of GGNBP2 were associated with poor prognosis in ESCC patients. Fbxo45 was negatively correlated with GGNBP2 expression in ESCC tissues. Therefore, Fbxo45 serves as an oncoprotein to promote ESCC tumorigenesis by targeting the stability of the tumor suppressor GGNBP2 in ESCC.
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Neoplasias Esofágicas , Carcinoma de Células Escamosas do Esôfago , Proteínas F-Box , Camundongos , Animais , Carcinoma de Células Escamosas do Esôfago/patologia , Neoplasias Esofágicas/patologia , Linhagem Celular Tumoral , Camundongos Nus , Ubiquitinação , Carcinogênese , Ubiquitina-Proteína Ligases/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas F-Box/genética , Proteínas F-Box/metabolismoRESUMO
Fbxo45, a conserved F-box protein, comprises of an atypical SKP1, CUL1, F-box protein (SCF) ubiquitin ligase complex that promotes tumorigenesis and development. However, the biological function and molecular mechanisms of Fbxo45 involved in pancreatic carcinogenesis are ambiguous. We conducted several approaches, including transfection, coIP, real-time polymerase chain reaction (RT-PCR), Western blotting, ubiquitin assays, and animal studies, to explore the role of Fbxo45 in pancreatic cancer. Here, we report that USP49 stability is governed by Fbxo45-mediated ubiquitination and is enhanced by the absence of Fbxo45. Moreover, Fbxo45 binds to a short consensus sequence of USP49 through its SPRY domain. Furthermore, Fbxo45-mediated USP49 ubiquitination and degradation are enhanced by NEK6 kinase. Functionally, Fbxo45 increases cell viability and motility capacity by targeting USP49 in pancreatic cancer cells. Xenograft mouse experiments demonstrated that ectopic expression of Fbxo45 enhanced tumor growth in mice and that USP49 overexpression inhibited tumor growth in vivo. Notably, Fbxo45 expression was negatively associated with USP49 expression in pancreatic cancer tissues. Fbxo45 serves as an oncoprotein to facilitate pancreatic oncogenesis by regulating the stability of the tumor suppressor USP49 in pancreatic cancer.