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Understanding the connection between senescence phenotypes and mitochondrial dysfunction is crucial in aging and premature aging diseases. Loss of mitochondrial function leads to a decline in T cell function, which plays a significant role in this process. However, more research is required to determine if improving mitochondrial homeostasis alleviates senescence phenotypes. Our research has shown an association between NAD+ and senescent T cells through the cGAS-STING pathway, which can lead to an inflammatory phenotype. Further research is needed to fully understand the role of NAD+ in T-cell aging and how it can be utilized to improve mitochondrial homeostasis and alleviate senescence phenotypes. We demonstrate here that mitochondrial dysfunction and cellular senescence with a senescence-associated secretory phenotype (SASP) occur in senescent T cells and tumor-bearing mice. Senescence is mediated by a stimulator of interferon genes (STING) and involves ectopic cytoplasmic DNA. We further show that boosting intracellular NAD+ levels with nicotinamide mononucleotide (NMN) prevents senescence and SASP by promoting mitophagy. NMN treatment also suppresses senescence and neuroinflammation and improves the survival cycle of mice. Encouraging mitophagy may be a useful strategy to prevent CD8+ T cells from senescence due to mitochondrial dysfunction. Additionally, supplementing with NMN to increase NAD+ levels could enhance survival rates in mice while also reducing senescence and inflammation, and enhancing mitophagy as a potential therapeutic intervention.
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Doenças Mitocondriais , NAD , Camundongos , Animais , NAD/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Mitocôndrias/metabolismo , Senescência Celular/fisiologia , Homeostase , Doenças Mitocondriais/metabolismo , Suplementos NutricionaisRESUMO
Many plants have the capability to accumulate anthocyanins for coloration, and anthocyanins are advantageous to human health. In the case of hulless barley (Hordeum vulgare L. var. nudum), investigation into the mechanism of anthocyanin formation is limited to the level of protein-coding genes (PCGs). Here, we conducted a comprehensive bioinformatics analysis to identify a total of 9414 long noncoding RNAs (lncRNAs) in the seed coats of purple and white hulless barley along a developmental gradient. Transcriptome-wide profiles of lncRNAs documented several properties, including GC content fluctuation, uneven length, a diverse range of exon numbers, and a wide variety of transcript classifications. We found that certain lncRNAs in hulless barley possess detectable sequence conservation with Hordeum vulgare and other monocots. Furthermore, both differentially expressed lncRNAs (DElncRNAs) and PCGs (DEPCGs) were concentrated in the later seed development stages. On the one hand, DElncRNAs could potentially cis-regulate DEPCGs associated with multiple metabolic pathways, including flavonoid and anthocyanin biosynthesis in the late milk and soft dough stages. On the other hand, there was an opportunity for trans-regulated lncRNAs in the color-forming module to affect seed coat color by upregulating PCGs in the anthocyanin pathway. In addition, the interweaving of hulless barley lncRNAs and diverse TFs may function in seed coat coloration. Notably, we depicted a dynamic portrait of the anthocyanin synthesis pathway containing hulless barley lncRNAs. Therefore, this work provides valuable gene resources and more insights into the molecular mechanisms underlying anthocyanin accumulation in hulless barley from the perspective of lncRNAs, which facilitate the development of molecular design breeding in crops.
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Hordeum , RNA Longo não Codificante , Antocianinas/genética , Antocianinas/metabolismo , Hordeum/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Sementes/genética , Tibet , TranscriptomaRESUMO
To enhance the characterization of wood extractives at molecular level, a detailed ultrahigh-resolution mass spectrometry (UHRMS)-based analytical methodology was developed in this work. The analytical strategies, including selection of compatible solvent for extraction, evaluation of ionization solvent for effective electrospray ionization, and multi-dimensional data analysis, were established to ensure a comprehensive characterization of complex compositions in wood extractives. Extraction capability of seven solvents with varied polarities was examined by a standard reference material of hardwood biomass and evaluated based on thousands of compounds which were much more than those discovered before. With a variety of data-processing approaches, including compound type distribution, double bond equivalent versus carbon number plot, and van Krevelen diagram, the chemodiversity of the extractives was fully explored from different perspectives. This work greatly expanded the compound library of wood extractives and could also provide guidance for the integrated composition analysis of other biomass materials.
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Madeira , Biomassa , Espectrometria de Massas , Solventes/química , Madeira/químicaRESUMO
BACKGROUND AND AIMS: Type 2 diabetes mellitus (T2DM) has high risk of developing cardiac dysfunction, increasing of either cardiovascular death or hospitalization for heart failure. MicroRNAs (miRNA) affect cardiac function of T2DM. The aim of this study was to investigate the relationships between five miRNA single nucleotide polymorphisms (SNP) and diastolic and systolic function of T2DM. METHODS AND RESULTS: Three hundred untreated T2DM subjects were included. Each subject underwent SNP genotyping, conventional echocardiography, tissue doppler imaging, and speckle tracking imaging. The effects of miRNA SNPs on diastolic and systolic function were evaluated. The diastolic function of T2DM subjects with miR-133a-1-rs8089787 wild genotype or let-7f-rs10877887 variant genotype was lower than those with miR-133a-1-rs8089787 variant genotype or let-7f-rs10877887 wild genotype, manifesting as higher left atrial volume index, lower mean E', and higher E/E' (P < 0.05). There were no significant effects of miR-133a-2-rs13040413, let-7a-1-rs13293512 and miR-27a-rs895819 on the diastolic function of T2DM subjects (P > 0.05). These five miRNA SNPs had no effect on the systolic function of T2DM subjects (P > 0.05). CONCLUSIONS: MiRNA-133a-1-rs8089787 and let-7f-rs10877887 were associated with impaired cardiac diastolic function in T2DM. The findings may be a promising therapeutic targets for preventing diastolic dysfunction in T2DM.
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Diabetes Mellitus Tipo 2 , MicroRNAs , Disfunção Ventricular Esquerda , Humanos , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/complicações , MicroRNAs/genética , Diástole , Polimorfismo de Nucleotídeo Único , Sístole , Disfunção Ventricular Esquerda/diagnóstico por imagem , Disfunção Ventricular Esquerda/genética , Função Ventricular EsquerdaRESUMO
Brassinolide (BR) is the "sixth class" plant hormone, which plays an important role in various physiological and biochemical processes of plants. The wide variety of functions of Pinellia ternata means that there is huge demand for it and thus it is in short supply. This paper mainly assessed the changes of yield and quality in P. ternata at different stages after BR treatments by principal component analysis, in order to improve the yield and quality of P. ternata and at the same time determine the best harvest time. The results showed that the tuber yield of P. ternata was significantly increased by BR treatments at different stages (except for the 15th day). After the 15th, 45th, 60th, 75th, 90th, and 105th day of treatments, the tuber yield of P. ternata reached peak values at 0.10 (0.65 g), 0.50 (1.97 g), 0.50 (1.98 g), 1.00 (2.37 g), 1.00 (2.84 g), and 2.00 mg/L (3.76 g) BR treatment, respectively. The optimal harvest time was the 75th day after 0.10, 0.50, and 1.00 mg/L BR treatments, which not only significantly improved the yield of P. ternata, but also retained high level of total alkaloids in the tubers (20.89, 5.37, and 13.44%) and bulbils (9.74, 20.42, and 13.62%), high total flavone content in the tubers (17.66, 16.26, and 12.74%) and bulbils (52.63, 12.79, and 38.69%), and high ß-sitosterol content in the tubers (25.26, 16.65, and 0.62%) of P. ternata, compared with the control, respectively.
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Alcaloides , Pinellia , Pinellia/química , Análise de Componente Principal , Tubérculos/química , Reguladores de Crescimento de Plantas/análise , Alcaloides/análiseRESUMO
CONTEXT: Saposhnikovia divaricata (Turcz.) Schischk (Apiaceae) (SD) has various pharmacological activities, but its effects on type I allergy (TIA) have not been comprehensively studied. OBJECTIVE: This study evaluates the treatment and molecular mechanisms of SD against TIA. MATERIALS AND METHODS: The effective components and action targets of SD were screened using TCMSP database, and allergy-related targets of SD were predicted using GeneCards and OMIM database. The obtained target intersections were imported into David database for GO analysis, and used R software to perform KEGG analysis. The RBL-2H3 cells sensitised by DNP-IgE/DNP-BSA were treated with different concentrations of SD (root decoction, 0.5, 1, and 2 mg/mL), prim-O-glucosylcimifugin (POG, 10, 40, and 80 µg/mL) and the positive control drug-ketotifen fumarate (KF, 30 µM) for 12 h, then subjected to cell degranulation and qPCR analysis. RESULTS: Eighteen active compounds of SD and 38 intersection targets were obtained: TIA-related signal pathways mainly include calcium signal pathway, PI3K-Akt signal pathway and MAPK signal pathway. Taking the ß-Hex release rate of the model group as the base, the release rate of SD and POG in high dose groups were 43.79% and 57.01%, respectively, which were significantly lower than model group (p < 0.01), and significantly lower than KF group (63.83%, p < 0.01, p < 0.05). SD and POG could down-regulate the expression of related proteins in the Lyn/Syk, PI3K/AKT and MAPK signalling pathways. DISCUSSION AND CONCLUSION: Saposhnikovia divaricata could inhibit IgE-induced degranulation of mast cells, providing a scientific basis for further research and clinical applications of SD in TIA treatment.
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Apiaceae , Medicamentos de Ervas Chinesas , Hipersensibilidade , Apiaceae/química , Medicamentos de Ervas Chinesas/farmacologia , Imunoglobulina E , Farmacologia em Rede , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-aktRESUMO
Thiophene compounds are the main concern of petroleum desulfurization, and their chemical composition and molecular configuration have critical impacts on thermodynamic and kinetic processes. In this work, atmospheric pressure chemical ionization (APCI) was employed for effective ionization of thiophene compounds in petroleum with complex matrix, in which carbon disulfide was used for generating predominant [M]+⢠ions without the need of derivatization as for electrospray ionization. APCI coupled with ultrahigh-resolution mass spectrometry (UHRMS) was successfully applied to the composition characterization of thiophene compounds in both a low boiling petroleum fraction and a whole crude oil. APCI coupled with trapped ion mobility spectrometry (TIMS) was developed to determine the shape and size of thiophene compounds, providing configuration information that affects the steric hindrance and diffusion behavior of reactants in the desulfurization reaction, which has not been previously reported. Moreover, the comprehensive experimental structural data, expressed as the collision cross section (CCS) of the ions as surrogates of molecules, provided clues to the factors affecting the desulfurization reactivity of thiophene compounds. Further exploration showed that not only qualitative analysis of thiophene compounds can be achieved from the correlation between m/z and CCS, but also molecular size was found to be correlated with CCS that can be used as structural analysis. Overall, the molecular composition and dimension analysis together can provide substantial information for the desulfurization activity of thiophene compounds, facilitating the desulfurization process studies and catalyst design.
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Aristolochic acid analogues (AAAs), naturally existing in herbal Aristolochia and Asarum genera, were once widely used in traditional pharmacopeias because of their anti-inflammatory properties, but lately they were identified as potential nephrotoxins and mutagens. A method for rapid characterization of AAAs in serum was developed using ion mobility spectrometry coupled with mass spectrometry (IMS-MS). Five AAAs, containing four aristolochic acids and one aristolactam, were separated and identified within milliseconds. AAAs were separated in gas phase based on the difference of their ion mobility (K0), and then identified based on their K0 values, m/z, and product ions from MS/MS. Quantitative analysis of AAAs was performed using an internal standard with a satisfactory sensitivity. Limits of detection (signal-to-noise = 3) and quantification (signal-to-noise = 10) were 1-5 ng/mL and 3-8 ng/mL, respectively. The method was validated and successfully applied to the pharmacokinetics study of AAAs in rats, offering a promising way for fast screening and evaluation of AAAs in biological samples.
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Ácidos Aristolóquicos/sangue , Animais , Aristolochia/química , Ácidos Aristolóquicos/química , Asarum/química , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacocinética , Espectrometria de Mobilidade Iônica/economia , Espectrometria de Mobilidade Iônica/métodos , Limite de Detecção , Masculino , Mutagênicos/química , Mutagênicos/farmacocinética , Ratos Sprague-DawleyRESUMO
OBJECTIVES: We designed a computer-based, integrated intelligent reminder system to reduce the deficiencies and errors in ultrasound (US) reports. In this study, we assessed the performance of this system and evaluated its impact on the quality of US reporting. METHODS: Ultrasound reporting deficiencies or errors were divided into 2 categories: missing items (including outpatient or inpatient number and clinical diagnosis) and content errors (including measurement data, sex-related, and laterality errors). The intelligent reminder system was designed in Visual Basic for Applications (Microsoft Corporation, Redmond, WA) and integrated with the US system. It automatically detects reporting errors before printing of the report and provides real-time prompts for correction of the errors. We compared the US reporting deficiencies and errors during the 20 months before and after implementation of the system. RESULTS: Before implementation of the system, deficiencies/errors were detected in 2.26% (8841 of 391,230) of US reports compared with 0.12% (530 of 444,215) of reports after implementation of the system (P < .0001). After adoption of the system, the reported item deficiencies were improved more than the content deficiencies, with the most notable improvement in clinical diagnosis. Sex-related errors were reduced from 7 cases to nil after use of the intelligent reminder system. No laterality errors were found before and after the implementation of the system. CONCLUSIONS: The intelligent reminder system within the US system significantly reduced deficiencies and errors, improving the quality of the report.
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Sistemas de Alerta , Humanos , UltrassonografiaRESUMO
Echocardiography is first-line examination of cardiac tumors. We report the case of a 25-year-old woman with a right atrial transmural invasive lipoma, and we review 58 published reports of primary cardiac invasive lipomas detected by echocardiography. We summarize the ultrasonographic characteristics and main sites of development, and examine the "invagination hypothesis". Echocardiography appears valuable for early detection, intraoperative monitoring, and postoperative follow-up of invasive cardiac lipomas.
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Neoplasias Cardíacas/diagnóstico por imagem , Lipoma/diagnóstico por imagem , Adulto , Ecocardiografia/métodos , Feminino , Átrios do Coração/diagnóstico por imagem , Átrios do Coração/patologia , Neoplasias Cardíacas/patologia , Neoplasias Cardíacas/cirurgia , Humanos , Lipoma/patologia , Lipoma/cirurgia , Invasividade Neoplásica , Resultado do TratamentoRESUMO
Electrolyte leakage is a severe safety concern in lithium batteries. With highly volatile 1,2-dimethoxyethane as solvent, the leakage related hazards are more pronounced in lithium-sulfur (Li-S) batteries. To address this concern, a leakage-proof electrolyte is delicately designed through functionalizing the commercial electrolyte by Li6 PS5 Cl-grafted poly(ethyl cyanoacrylate), which can interact readily with the aluminum-plastic packing through hydrogen bond to immobilize the electrolyte. The moisture from ambient can also catalyze a further polymerization of the macromolecules to seal the leaking points and thereby to solve the leakage issue, endowing Li-S batteries superior safety even in an artificial cut pouch cell. With a bare S loading of 4.9â mg cm-2 , the battery can deliver good endurance owing to the suppressed polysulfide shuttle by its polar groups. This work enlightens the design of leakage-proof electrolyte and makes a milestone for high performance Li-S batteries.
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Microbes, especially the uncultured microbes, have been considered as an important resource for discovery of novel cellulases. In this study, a novel bifunctional cellulase/hemicellulase (ZFYN184) was identified by functional screening of a soil metagenomic library. Sequence analysis indicated that ZFYN184 shared at best 39% identity with glycoside hydrolase family 44 (GH44) proteins and contained a glutamic acid residue at 235 acting as the catalytic proton donor in hydrolysis of polysaccharides. The recombinant ZFYN184 was expressed in Escherichia coli BL21 (DE3), and the biochemical profiles of the enzyme, including optimum pH and temperature, pH and thermal stabilities, tolerance to various additives, and substrate specificity, were determined. ZFYN184 possessed strong endo-ß-1,4-glucanase and endo-1,4-ß-mannanase activities, as well as weak xylanase activity, while all these hydrolytic activities were derived from a single catalytic domain in this GH44 enzyme. KEY POINTS: ⢠Discovery a novel bifunctional glycosyl hydrolase from uncultured microorganism. ⢠ZFYN184 contains a single catalytic domain belonged to GH44.
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Celulase , Celulases , Celulase/genética , Celulase/metabolismo , Clonagem Molecular , Estabilidade Enzimática , Glicosídeo Hidrolases/metabolismo , Concentração de Íons de Hidrogênio , Solo , Especificidade por SubstratoRESUMO
Diabetes mellitus (DM) has become the third major chronic non-communicable disease affecting global public health, following cancer and cardiovascular and cerebrovascular diseases. Although previous studies have found a correlation between microRNA (miRNA) and the development of DM, thus far, most reviews have focused on the studies describing the changes in miRNA expression profiles and the mechanisms by which miRNAs-induce DM. However, reviews summarizing the effect of miRNA single-nucleotide polymorphisms on the developmental stages of DM and its complications are still needed. Studies on the variation of miRNAs will not only help to further understand the role that genetic factors play in DM, but will also have important implications for treatment and disease prognosis.
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Diabetes Mellitus/genética , Predisposição Genética para Doença , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único , Animais , Autoimunidade , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/metabolismo , Regulação da Expressão Gênica , Estudos de Associação Genética , Humanos , Insulina/metabolismo , Resistência à Insulina , Interferência de RNARESUMO
The cyclic Arg-Gly-Asp (cRGD) peptides are widely used as tumor-targeting ligands due to their specific binding ability to integrin αvß3, which is overexpressed on the surface of various cancer cells and the endothelial cells of new blood vessels within tumor tissues. In this paper, the postinsertion strategy of DSPE-PEG2000-cRGD has been applied to the nanoparticles of 3',3â³-bis-peptide-siRNA (pp-siRNA) encapsulated by gemini-like cationic lipid (CLD) and neutral cytosin-1-yl lipid (DNCA) from our lab. It was confirmed that the nanoparticles of pp-siRNA/CLD/DNCA/DSPE-PEG2000-cRGD (PCNR) were able to specifically target tumor cells with highly expressed integrin αvß3; moreover, it efficiently downregulated the levels of BRAF mRNA and the BRAF protein and inhibited cell proliferation in A375 cells, in comparison with the nontargeted nanocomplex of pp-siRNA/CLD/DNCA/cRAD (PCNA). The uptake pathways of PCNR are mostly dependent on CvME-mediated endocytosis and macropinocytosis in A375 cells, which could bypass lysosome or quickly lead to the lysosomal escape to reduce siRNA degradation. Finally, the biodistribution study showed that PCNR exhibited a high ability to accumulate in tumor tissues. These results suggest that the nanocomplex of PCNR is promising to be highly effective in the treatment of melanomas including their mutation.
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Nanopartículas/química , Peptídeos Cíclicos/química , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Células A549 , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Células HeLa , Humanos , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Monocyte chemoattractant protein-induced protein 1 (MCPIP1) is a kind of zinc finger RNA binding protein, which exerts immune responses in a variety of cell types. However, the role of MCPIP1 in bovine mammary epithelial cells during mastitis has not been studied. In this study, we explored the functions of MCPIP1 in the inflammatory process induced by virulence factors of pathogens in bovine mammary alveolar cell-T (MAC-T) cell line. Our results showed that MCPIP1 was significantly highly expressed both in the mammary tissue of dairy cows with mastitis and in inflammatory MAC-T cells induced by lipopolysaccharide (LPS) or lipoteichoic acid (LTA). Furthermore, we found that overexpression of MCPIP1 in MAC-T cells abated the LPS-induced increase at the gene expression levels of inflammatory mediators tumor necrosis factor-α-α, interleukin (IL)-1ß, IL-6 and IL-8, enhanced the LPS- and LTA-induced inhibition of epithelial proliferation and promoted the LPS- and LTA-induced oxidative and DNA damage. These findings indicated that MCPIP1 has an enormous potential in regulating the inflammatory response of bovine mammary epithelial cells during infection and may provide an effective therapeutic target for bovine mastitis to reduce the damage caused by inflammatory reactions.
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Citocinas/imunologia , Células Epiteliais/imunologia , Mediadores da Inflamação/imunologia , Mastite Bovina/imunologia , Proteínas de Ligação a RNA/metabolismo , Dedos de Zinco , Animais , Bovinos , Linhagem Celular , Proliferação de Células/genética , Citocinas/genética , Citocinas/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos , Glândulas Mamárias Animais/citologia , Mastite Bovina/genética , Mastite Bovina/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/imunologia , Proteínas de Ligação a RNA/genética , Ácidos TeicoicosRESUMO
The interaction of immunoglobulin E (IgE) to its high-affinity receptor (FcεRI) plays a key role in triggering allergic reactions. However, it is still controversial whether species specificity influences the binding ability between humans and rodents. Recombinant hFcεRIα / RBL-2H3 and hFcεRIα / MGC-803 cells were prepared and sensitized with hIgE/anti-hIgE antibody or DNP-IgE/DNP-BSA, respectively. Species-specificity was investigated using immuno-fluorescent analysis, ß-hexosaminidase release rate assay, intracellular calcium concentration assay and apoptosis assay, and the results showed that there is species-specificity in IgE/FcεRI in humans and rats, and no cross-recognition between IgE and FcεRI in two species. These results should provide the experimental evidence for further research on the pathogenesis and the drug development of allergic diseases.
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Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Receptores de IgE/imunologia , Animais , Apoptose/imunologia , Cálcio/metabolismo , Linhagem Celular Tumoral , Imunofluorescência , Humanos , Ratos , Especificidade da EspécieRESUMO
Allergic asthma is a chronic inflammatory disease and involves many cells and cellular components. Cataract is a condition that affects the transparency of the lens, which the opacity of the lens caused by any innate or acquired factor degrades its transparency or changes in color. During the establishment of asthma model of rats with chicken ovalbumin nebulization, it was found that asthmatic rats were more likely to have monocular or binocular cataract symptoms than normal rats. Considering that they are all induced by immune imbalance, inflammation, etc., there may be some correlation in the mechanism, and many clues showed that both diseases are associated with activation of the PI3K-AKT-mTOR signaling pathway. Therefore, we hypothesized that the PI3K-AKT-mTOR signaling pathway produces inflammatory or immune imbalance based on allergy leading to cataract.
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Asma/induzido quimicamente , Catarata/induzido quimicamente , Animais , Modelos Animais de Doenças , Hipersensibilidade , Inflamação , Masculino , Ovalbumina , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Wistar , Serina-Treonina Quinases TOR/metabolismoRESUMO
Mud volcanoes (MVs) emit globally significant quantities of methane into the atmosphere, however, methane cycling in such environments is not yet fully understood, as the roles of microbes and their associated biogeochemical processes have been largely overlooked. Here, we used data from high-throughput sequencing of microbial 16S rRNA gene amplicons from six MVs in the Junggar Basin in northwest China to quantify patterns of diversity and characterize the community structure of archaea and bacteria. We found anaerobic methanotrophs and diverse sulfate- and iron-reducing microbes in all of the samples, and the diversity of both archaeal and bacterial communities was strongly linked to the concentrations of sulfate, iron and nitrate, which could act as electron acceptors in anaerobic oxidation of methane (AOM). The impacts of sulfate/iron/nitrate on AOM in the MVs were verified by microcosm experiments. Further, two representative MVs were selected to explore the microbial interactions based on phylogenetic molecular ecological networks. The sites showed distinct network structures, key species and microbial interactions, with more complex and numerous linkages between methane-cycling microbes and their partners being observed in the iron/sulfate-rich MV. These findings suggest that electron acceptors are important factors driving the structure of microbial communities in these methane-rich environments.
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Archaea/metabolismo , Bactérias/metabolismo , Elétrons , Metano/metabolismo , Microbiota , Microbiologia do Solo , Anaerobiose , Archaea/genética , Bactérias/genética , Biodiversidade , China , Transporte de Elétrons , Sedimentos Geológicos/microbiologia , Fenômenos Geológicos , Oxirredução , RNA Ribossômico 16S/genéticaRESUMO
Recently, a new oxygenic pathway has been proposed based on the disproportionation of NO with putative NO dismutase (Nod). In addition to a new process in nitrogen cycling, this process provides ecological advantages for the degradation of substrates in anaerobic conditions, which is of great significance for wastewater treatment. However, the Nod distribution in aquatic environments is rarely investigated. In this study, we obtained the nod genes with an abundance of 2.38 ± 0.96 × 105 copies per gram of dry soil from the Zoige wetland and aligned the molecular characteristics in the corresponding Nod sequences. These Nod sequences were not only found existing in NC10 bacteria, but were also found forming some other clusters with Nod sequences from a WWTP reactor or contaminated aquifers. Moreover, a new subcluster in the aquifer-similar cluster was even dominant in the Zoige wetland and was named the Z-aquifer subcluster. Additionally, soils from the Zoige wetland showed a high potential rate (10.97 ± 1.42 nmol of CO2 per gram of dry soil per day) for nitrite-dependent anaerobic methane oxidation (N-DAMO) with low abundance of NC10 bacteria, which may suggest a potential activity of Nod in other clusters when considering the dominance of the Z-aquifer subcluster Nod. In conclusion, we verified the occurrence of Nod in an alpine wetland for the first time and found a new subcluster to be dominant in the Zoige wetland. Moreover, this new subcluster of Nod may even be active in the N-DAMO process in this alpine wetland, which needs further study to confirm.
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Bactérias Anaeróbias/enzimologia , Bactérias Anaeróbias/metabolismo , Microbiologia Ambiental , Metano/metabolismo , Óxido Nítrico/metabolismo , Áreas Alagadas , Anaerobiose , Metagenoma , Análise de Sequência de DNARESUMO
IgE/FcεRI signal pathway plays a crucial role in triggering allergic reactions, and there is no cross-recognition between IgE and FcεRI in human and rats. In order to obtain the hFcεRIα/ RBL-2H3 cell line, total RNA was extracted from U937 cells, and the human FcεRIα gene was obtained by RT-PCR technology. Then the amplified product was digested and inserted into the pIRES2-EGFP vector. After the plasmid was transfected into the RBL-2H3 cells using lipofectamine, and the RBL-2H3 cell lines of stable expression were screened by G418. The transfection efficiency reached 60.45% with optimizing transfection parameters. The last the expression of hFcεRIα was detected by RT-PCR, western blotting and fluorescent microscopy. The present results demonstrated that the pIRES2-EGFP-hFcεRIα vector was constructed and a stable cell line of hFcεRIα/ RBL-2H3 cells was established successfully. This cell line is promising tools for further research on the pathogenesis and drug development of allergic diseases.