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1.
Mol Cell Proteomics ; : 100837, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-39243857

RESUMO

Universal sample preparation for proteomic analysis that enables unbiased protein manipulation, flexible reagent use, and low protein loss is required to ensure the highest sensitivity of downstream liquid chromatography-mass spectrometry (LC-MS) analysis. To address these needs, we developed a ZnCl2 precipitation-assisted sample preparation method (ZASP) that depletes harsh detergents and impurities in protein solutions prior to trypsin digestion via 10 min of ZnCl2 and methanol-induced protein precipitation at room temperature (RT). ZASP can remove trypsin digestion and LC-MS incompatible detergents such as SDS, Triton X-100, and urea at high concentrations in solution and unbiasedly recover proteins independent of the amount of protein input. We demonstrated the sensitivity and reproducibility of ZASP in an analysis of samples with 1 µg to 1000 µg of proteins. Compared to commonly used sample preparation methods such as SDC-based in-solution digestion, acetone precipitation, FASP, and SP3, ZASP has proven to be an efficient approach. Here, we present ZASP, a practical, robust, and cost-effective proteomic sample preparation method that can be applied to profile different types of samples.

2.
Mol Cell ; 66(5): 711-720.e3, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28529057

RESUMO

The multidomain CCCTC-binding factor (CTCF), containing a tandem array of 11 zinc fingers (ZFs), modulates the three-dimensional organization of chromatin. We crystallized the human CTCF DNA-binding domain in complex with a known CTCF-binding site. While ZF2 does not make sequence-specific contacts, each finger of ZF3-7 contacts three bases of the 15-bp consensus sequence. Each conserved nucleotide makes base-specific hydrogen bonds with a particular residue. Most of the variable base pairs within the core sequence also engage in interactions with the protein. These interactions compensate for deviations from the consensus sequence, allowing CTCF to adapt to sequence variations. CTCF is sensitive to cytosine methylation at position 2, but insensitive at position 12 of the 15-bp core sequence. These differences can be rationalized structurally. Although included in crystallizations, ZF10 and ZF11 are not visible, while ZF8 and ZF9 span the backbone of the DNA duplex, conferring no sequence specificity but adding to overall binding stability.


Assuntos
Metilação de DNA , DNA/metabolismo , Proteínas Repressoras/metabolismo , 5-Metilcitosina/metabolismo , Sítios de Ligação , Fator de Ligação a CCCTC , Clonagem Molecular , Cristalografia por Raios X , DNA/química , DNA/genética , Humanos , Ligação de Hidrogênio , Modelos Moleculares , Conformação de Ácido Nucleico , Ligação Proteica , Estabilidade Proteica , Proteínas Repressoras/química , Proteínas Repressoras/genética , Relação Estrutura-Atividade , Repetições de Trinucleotídeos , Dedos de Zinco
3.
Arch Insect Biochem Physiol ; 115(1): e22086, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38288496

RESUMO

The mitochondrial gene order in Thysanoptera is notably distinct and highly rearranged, with each species exhibiting its own unique arrangement. To elucidate the relationship between gene rearrangements and phylogeny, the complete mitochondrial genome (mitogenome) of the wheat pest, Aptinothrips stylifer, was sequenced and assembled, spanning a total length of 16,033 bp. Compared with the ancestral arthropod mitogenome, significant rearrangement differences were evident in A. stylifer, whereas the gene order between A. stylifer and Anaphothrips obscurus was similar. Phylogenetic trees were reconstructed based on all 13 protein-coding gene sequences using Bayesian inference and maximum-likelihood methods, both yielding similar topological structures. Notably, A. stylifer was robustly clustered with A. obscurus, affirming its classification within Anaphothrips genus group. This exemplifies the potential correlation between gene rearrangements and phylogeny in the Thripidae family. Additionally, the mitogenome of A. stylifer exhibited several atypical features, including: (1) Three putative control regions (CRs) in close proximity, with CR2 and CR3 displaying partial similarity, and CR1 differing in base composition; (2) Two transfer RNAs (tRNAs), trnS1 and trnV, lacking the DHU arm; (3) Two ribosomal RNA (rRNA) genes inverted and positioned distant from each other; (4) Negative AT and GC skew (AT skew = -0.001, GC skew = -0.077); (5) One transposition (nad6), one inverse transposition (trnQ), four inversions (trnF, trnH, trnC, and gene block nad1-trnL1-rrnL-trnV-rrnS), and four tandem duplication random loss events; and (6) Two protein-coding genes, nad2 and atp8, terminated with an incomplete stop codon "T".


Assuntos
Genoma Mitocondrial , Tisanópteros , Animais , Filogenia , Tisanópteros/genética , Triticum/genética , Teorema de Bayes
4.
BMC Nephrol ; 25(1): 81, 2024 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-38443857

RESUMO

OBJECTIVE: To validate an association between new inflammation and frequent peritoneal dialysis-associated peritonitis (PDAP). MATERIALS AND METHODS: In China, retrospective clinical data were collected on 208 patients who received continuous ambulatory peritoneal dialysis (CAPD) between 2010 and 2021. The patients were divided into two groups: non-frequent PDAP (the interval between two peritonitis episodes of more than one year) and frequent PDAP (the interval between two peritonitis episodes of less than one year). Patients with their first episode of peritonitis had their age, gender, history of hypertension, diabetic disease, underlying renal disease, bacterial infection, and laboratory data collected. The outcomes of bacterial dispersion, systemic immune-inflammation index (SII), high-density lipoprotein cholesterol (HDL-C), C-reactive protein (CRP), and risk variables associated with frequent PDAP were analyzed. RESULTS: There are differences between the two groups in dialysis time (p = 0.006), hypertensive nephropathy (p = 0.038), staphylococcus (p = 0.035), white blood cells (p = 0.001), neutrophil (p < 0.01), lymphocyte (p < 0.01), platelet(p = 0.01), SII(p < 0.01), CRP/HDL-C (p = 0.002), CRP (p < 0.001), serum creatinine (p = 0.007), blood urea nitrogen (p = 0.05), serum magnesium (0.03), serum potassium (p = 0.007), and dialysate polymorphonuclear cells (p = 0.004). Multifactorial logistic regression analysis found that SII (p < 0.001), CRP/HDL-C (p = 0.041), and Diabetes mellitus (p = 0.027) were independent risk factors for frequent PDAP. The ROC curve analysis revealed that combining SII with CRP/HDL-C resulted in the largest AUC area (AUC = 0.814). CONCLUSIONS: Our findings offer clinical proof of the combination of SII and CRP/HDL-C in patients with frequent PDAP.


Assuntos
Diálise Peritoneal , Peritonite , Humanos , Estudos Retrospectivos , Diálise Renal , Inflamação/etiologia , Peritonite/epidemiologia , Peritonite/etiologia , Diálise Peritoneal/efeitos adversos , HDL-Colesterol
5.
Food Microbiol ; 120: 104467, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38431319

RESUMO

The luxS mutant strains of Shewanella putrefaciens (SHP) were constructed to investigate the regulations of gene luxS in spoilage ability. The potential regulations of AI-2 quorum sensing (QS) system and activated methyl cycle (AMC) were studied by analyzing the supplementation roles of key circulating substances mediated via luxS, including S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), methionine (Met), homocysteine (Hcy) and 4,5-dihydroxy-2,3-pentanedione (DPD). Growth experiments revealed that the luxS deletion led to certain growth limitations of SHP, which were associated with culture medium and exogenous additives. Meanwhile, the decreased biofilm formation and diminished hydrogen sulfide (H2S) production capacity of SHP were observed after luxS deletion. The relatively lower total volatile base nitrogen (TVB-N) contents and higher sensory scores of fish homogenate with luxS mutant strain inoculation also indicated the weaker spoilage-inducing effects after luxS deletion. However, these deficiencies could be offset with the exogenous supply of circulating substances mentioned above. Our findings suggested that the luxS deletion would reduce the spoilage ability of SHP, which was potentially attributed to the disorder of AMC and AI-2 QS system.


Assuntos
Percepção de Quorum , Shewanella putrefaciens , Animais , Percepção de Quorum/genética , Shewanella putrefaciens/genética , Shewanella putrefaciens/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Metionina/genética , Metionina/metabolismo , Biofilmes , Regulação Bacteriana da Expressão Gênica
6.
Sensors (Basel) ; 24(9)2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38732797

RESUMO

Flatness is a critical parameter in the manufacturing industry, directly impacting the fit and overall product performance. As the efficiency of manufacturing continues to advance, there is an increasing demand for more accurate and efficient measurement techniques. Existing methods often struggle to strike a balance between precision and efficiency. In response, this article introduces a novel approach that is capable of achieving high-precision and rapid measurements concerning multiple surfaces. By enhancing the traditional phase measuring deflectometry (PMD) method, employing a matching technique based on polar lines and normal vector constraints to address discrete surface measurement challenges, and implementing a plane pre-positioning method to tackle low efficiency in binocular matching and solving, we successfully performed swift and synchronized measurements for a large batch of specular surfaces and obtained the three-dimensional surface profile of each measured surface. Through experimental validation, the method proposed in this paper can perform the batch measurement of specular planes while maintaining high measurement accuracy.

7.
Molecules ; 29(9)2024 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-38731489

RESUMO

Gallic acid (GA) is a type of polyphenolic compound that can be found in a range of fruits, vegetables, and tea. Although it has been confirmed it improves non-alcoholic fatty liver disease (NAFLD), it is still unknown whether GA can improve the occurrence of NAFLD by increasing the low-density lipoprotein receptor (LDLR) accumulation and alleviating cholesterol metabolism disorders. Therefore, the present study explored the effect of GA on LDLR and its mechanism of action. The findings indicated that the increase in LDLR accumulation in HepG2 cells induced by GA was associated with the stimulation of the epidermal growth factor receptor-extracellular regulated protein kinase (EGFR-ERK1/2) signaling pathway. When the pathway was inhibited by EGFR mab cetuximab, it was observed that the activation of the EGFR-ERK1/2 signaling pathway induced by GA was also blocked. At the same time, the accumulation of LDLR protein and the uptake of LDL were also suppressed. Additionally, GA can also promote the accumulation of forkhead box O3 (FOXO3) and suppress the accumulation of hepatocyte nuclear factor-1α (HNF1α), leading to the inhibition of proprotein convertase subtilisin/kexin 9 (PCSK9) mRNA expression and protein accumulation. This ultimately results in increased LDLR protein accumulation and enhanced uptake of LDL in cells. In summary, the present study revealed the potential mechanism of GA's role in ameliorating NAFLD, with a view of providing a theoretical basis for the dietary supplementation of GA.


Assuntos
Ácido Gálico , Lipoproteínas LDL , Receptores de LDL , Humanos , Ácido Gálico/farmacologia , Receptores de LDL/metabolismo , Células Hep G2 , Lipoproteínas LDL/metabolismo , Receptores ErbB/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Pró-Proteína Convertase 9/metabolismo , Pró-Proteína Convertase 9/genética
8.
Angew Chem Int Ed Engl ; : e202417362, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-39278829

RESUMO

Imide functionalization has been widely proved to be an effective approach to enrich optoelectronic properties of polycyclic aromatic hydrocarbons (PAHs). However, appending multiple imide groups onto linear acenes is still a synthetic challenge. Herein, we demonstrate that by taking advantage of a "breaking and mending" strategy, a linear pentacene tetraimides (PeTI) was synthesized through a three-step sequence started from the naphthalene diimides (NDI). Compared with the parent pentacene, PeTI shows a deeper-lying lowest unoccupied molecular orbital (LUMO) energy level, narrower bandgap and better stability. The redox behavior of PeTI was firstly evaluated by generating a stable radical anion specie with the assistance of cobaltocene (CoCp2), and the structure of the electron transfer (ET) complex was confirmed by the X-ray crystallography. Moreover, due to the presence of multiple redox-active sites, we are able to show that the state-of-the-art energy storage performance of the dealkylated PeTI (designated as PeTCTI) in organic potassium ion batteries (OPIBs) as an anode. Our results shed light on the application of multiple imides functionalized linear acenes, and the reported synthetic strategy provides an effective way to get access to longer nanoribbon imides with fascinating electronic properties.

9.
BMC Genomics ; 24(1): 540, 2023 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-37700222

RESUMO

BACKGROUND: Intramuscular fat (IMF) content is the major indicator for evaluating chicken meat quality due to its positive correlation with tenderness, juiciness, and flavor. An increasing number of studies are focusing on the functions of microRNAs (miRNAs) in intramuscular adipocyte differentiation. However, little is known about the association of miR-128-3p with intramuscular adipocyte differentiation. Our previous RNA-seq results indicated that miR-128-3p was differentially expressed at different periods in chicken intramuscular adipocytes, revealing a possible association with intramuscular adipogenesis. The purpose of this research was to investigate the biological functions and regulatory mechanism of miR-128-3p in chicken intramuscular adipogenesis. RESULTS: The results of a series of assays confirmed that miR-128-3p could promote the proliferation and inhibit the differentiation of intramuscular adipocytes. A total of 223 and 1,050 differentially expressed genes (DEGs) were identified in the mimic treatment group and inhibitor treatment group, respectively, compared with the control group. Functional enrichment analysis revealed that the DEGs were involved in lipid metabolism-related pathways, such as the MAPK and TGF-ß signaling pathways. Furthermore, target gene prediction analysis showed that miR-128-3p can target many of the DEGs, such as FDPS, GGT5, TMEM37, and ASL2. The luciferase assay results showed that miR-128-3p targeted the 3' UTR of FDPS. The results of subsequent functional assays demonstrated that miR-128-3p acted as an inhibitor of intramuscular adipocyte differentiation by targeting FDPS. CONCLUSION: miR-128-3p inhibits chicken intramuscular adipocyte differentiation by downregulating FDPS. Our findings provide a theoretical basis for the study of lipid metabolism and reveal a potential target for molecular breeding to improve meat quality.


Assuntos
Galinhas , MicroRNAs , Animais , Galinhas/genética , Diferenciação Celular/genética , Adipogenia/genética , Regiões 3' não Traduzidas , Adipócitos , MicroRNAs/genética
10.
J Hepatol ; 78(4): 805-819, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36669703

RESUMO

BACKGROUND & AIMS: Capsaicin receptor, also known as transient receptor potential vanilloid 1 (TRPV1), is involved in pain physiology and neurogenic inflammation. Herein, we discovered the presence of TRPV1 in hepatic stellate cells (HSCs) and aimed to delineate its function in this cell type and liver fibrosis. METHODS: TRPV1 expression was examined in liver biopsies from patients with liver fibrosis using quantitative real-time PCR and immunostaining. Its contribution to liver fibrosis was examined in Trpv1-/- mice, upon lentiviral delivery of the TRPV1 gene, and in human and mouse primary HSCs, using patch clamp, intracellular Ca2+ mobilization determination, FACS analyses and gain/loss of function experiments. Binding of sterile alpha and Toll/interleukin-1 receptor motif-containing protein 1 (SARM1) to TRPV1 was determined using mass spectrometry, co-immunoprecipitation, surface plasmon resonance, bioluminescence resonance energy transfer, and NanoBiT. RESULTS: TRPV1 mRNA levels are significantly downregulated in patients with liver fibrosis and mouse models, showing a negative correlation with F stage and α-smooth muscle actin expression, a marker of HSC activation. TRPV1 expression and function decrease during HSC activation in fibrotic livers in vivo or during culture. Genetic and pharmacological inhibition of TRPV1 in quiescent HSCs leads to NF-κB activation and pro-inflammatory cytokine production. TRPV1 requires binding of its N-terminal ankyrin repeat domain to the TIR-His583 (Toll/interleukin-1 receptor) domain of SARM1 to prevent HSCs from pro-inflammatory activation. Trpv1-/- mice display increased HSC activation and more severe liver fibrosis, whereas TRPV1 overexpression is antifibrotic in various disease models. CONCLUSION: The antifibrotic properties of TRPV1 are attributed to the prevention of HSC activation via the recruitment of SARM1, which could be an attractive therapeutic strategy against liver fibrosis. IMPACT AND IMPLICATIONS: We identified the neuronal channel protein TRPV1 as a gatekeeper of quiescence in hepatic stellate cells, a key driver of liver fibrogenesis and chronic liver disease. Physiologically expressed in healthy liver and consistently downregulated during liver fibrosis development, its therapeutic re-expression is expected to have few side effects, making it an attractive target diagnostic tool and drug candidate for industry and clinicians.


Assuntos
Células Estreladas do Fígado , Canais de Cátion TRPV , Humanos , Camundongos , Animais , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Canais de Cátion TRPV/farmacologia , Células Estreladas do Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/patologia , Regulação da Expressão Gênica , Proteínas do Citoesqueleto/metabolismo , Proteínas do Citoesqueleto/farmacologia , Proteínas do Domínio Armadillo/genética , Proteínas do Domínio Armadillo/metabolismo
11.
Anal Chem ; 95(49): 17974-17980, 2023 12 12.
Artigo em Inglês | MEDLINE | ID: mdl-38011496

RESUMO

Global phosphoproteome profiling can provide insights into cellular signaling and disease pathogenesis. To achieve comprehensive phosphoproteomic analyses with minute quantities of material, we developed a rapid and sensitive phosphoproteomics sample preparation strategy based on ultrasound. We found that ultrasonication-assisted digestion can significantly improve peptide identification by 20% due to the generation of longer peptides that can be detected by mass spectrometry. By integrating this rapid ultrasound-assisted peptide-identification-enhanced proteomic method (RUPE) with streamlined phosphopeptide enrichment steps, we established RUPE-phospho, a fast and efficient strategy to characterize protein phosphorylation in mass-limited samples. This approach dramatically reduces the sample loss and processing time: 24 samples can be processed in 3 h; 5325 phosphosites, 4549 phosphopeptides, and 1888 phosphoproteins were quantified from 5 µg of human embryonic kidney (HEK) 293T cell lysate. In addition, 9219 phosphosites were quantified from 1-2 mg of OCT-embedded mouse brain with 120 min streamlined RUPE-phospho workflow. RUPE-phospho facilitates phosphoproteome profiling for microscale samples and will provide a powerful tool for proteomics-driven precision medicine research.


Assuntos
Fosfoproteínas , Proteômica , Animais , Camundongos , Humanos , Proteômica/métodos , Fluxo de Trabalho , Fosforilação , Fosfoproteínas/metabolismo , Fosfopeptídeos/análise , Proteoma/metabolismo
12.
PLoS Pathog ; 17(4): e1009561, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33905456

RESUMO

The H7N9 avian influenza virus (AIV) that emerged in China have caused five waves of human infection. Further human cases have been successfully prevented since September 2017 through the use of an H7N9 vaccine in poultry. However, the H7N9 AIV has not been eradicated from poultry in China, and its evolution remains largely unexplored. In this study, we isolated 19 H7N9 AIVs during surveillance and diagnosis from February 2018 to December 2019, and genetic analysis showed that these viruses have formed two different genotypes. Animal studies indicated that the H7N9 viruses are highly lethal to chicken, cause mild infection in ducks, but have distinct pathotypes in mice. The viruses bound to avian-type receptors with high affinity, but gradually lost their ability to bind to human-type receptors. Importantly, we found that H7N9 AIVs isolated in 2019 were antigenically different from the H7N9 vaccine strain that was used for H7N9 influenza control in poultry, and that replication of these viruses cannot, therefore, be completely prevented in vaccinated chickens. We further revealed that two amino acid mutations at positions 135 and 160 in the HA protein added two glycosylation sites and facilitated the escape of the H7N9 viruses from the vaccine-induced immunity. Our study provides important insights into H7N9 virus evolution and control.


Assuntos
Subtipo H7N9 do Vírus da Influenza A/genética , Subtipo H7N9 do Vírus da Influenza A/isolamento & purificação , Vacinas contra Influenza/uso terapêutico , Influenza Aviária/prevenção & controle , Doenças das Aves Domésticas/virologia , Animais , Animais de Zoológico/virologia , Galinhas/virologia , China/epidemiologia , Patos/virologia , Controle de Infecções/métodos , Subtipo H7N9 do Vírus da Influenza A/classificação , Subtipo H7N9 do Vírus da Influenza A/fisiologia , Influenza Aviária/epidemiologia , Influenza Aviária/virologia , Camundongos , Filogenia , Vigilância da População , Aves Domésticas , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/prevenção & controle
13.
Cell Mol Neurobiol ; 43(1): 395-408, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35152327

RESUMO

Microglia are the main immune cells of the central nervous system (CNS) and comprise various model systems used to investigate inflammatory mechanisms in CNS disorders. Currently, shaking and mild trypsinization are widely used microglial culture methods; however, the problems with culturing microglia include low yield and a time-consuming process. In this study, we replaced normal culture media (NM) with media containing 25% fibroblast-conditioned media (F-CM) to culture mixed glia and compared microglia obtained by these two methods. We found that F-CM significantly improved the yield and purity of microglia and reduced the total culture time of mixed glia. The microglia obtained from the F-CM group showed longer ramified morphology than those from the NM group, but no difference was observed in cell size. Microglia from the two groups had similar phagocytic function and baseline phenotype markers. Both methods yielded microglia were responsive to various stimuli such as lipopolysaccharide (LPS), interferon-γ (IFN-γ), and interleukin-4 (IL-4). The current results suggest that F-CM affect the growth of primary microglia in mixed glia culture. This method can produce a high yield of primary microglia within a short time and may be a convenient method for researchers to investigate inflammatory mechanisms and some CNS disorders.


Assuntos
Microglia , Neuroglia , Meios de Cultivo Condicionados/farmacologia , Células Cultivadas , Fibroblastos , Lipopolissacarídeos/farmacologia
14.
Support Care Cancer ; 32(1): 12, 2023 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-38060050

RESUMO

PURPOSE: High output stoma(HOS) is one of the most common complications after ileostomy, leading to fluid and electrolyte disturbances and renal dysfunction, and increasing the risk of readmission. Routine health education for HOS should be provided, and nurses, as the primary educators, should have adequate knowledge and skills in this area. However, there is a paucity of research on the knowledge and practice of HOS management. This study used the Knowledge, Attitude and Behavioural Practice Model to assess the management of HOS by colorectal surgery nurses and to explore the factors that influence it. METHOD: Using a multi-centre, cross-sectional study design, 398 colorectal surgery nurses from 6 hospitals in 6 cities in 6 provinces were surveyed using a structured electronic questionnaire to assess general information and knowledge, attitudes and management practices and training needs related to HOS of ileostomy. RESULTS: Colorectal surgery nurses' knowledge and practice of HOS was low. The presence or absence of training is an important factor influencing nurses' knowledge, attitudes and practice, with most nurses having no training and stoma specialist nurses scoring relatively high on knowledge and practice. CONCLUSIONS: Nurses play a very important role in the management of HOS, but this study shows that the current level of knowledge and practice of HOS among colorectal surgery nurses is concerning, and whether or not they have received training is the most critical influencing factor; therefore, training related to HOS is urgently needed.


Assuntos
Neoplasias Colorretais , Enfermeiras e Enfermeiros , Humanos , Ileostomia , Estudos Transversais , Conhecimentos, Atitudes e Prática em Saúde , Competência Clínica , Inquéritos e Questionários , Neoplasias Colorretais/cirurgia , Atitude do Pessoal de Saúde
15.
Mol Cell ; 57(2): 376-88, 2015 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-25557549

RESUMO

Posttranscriptional regulation in eukaryotes requires cis- and trans-acting features and factors including RNA secondary structure and RNA-binding proteins (RBPs). However, a comprehensive view of the structural and RBP interaction landscape of nuclear RNAs has yet to be compiled for any organism. Here, we use our ribonuclease-mediated structure and RBP-binding site mapping approaches to globally profile these features in Arabidopsis seedling nuclei in vivo. We reveal anticorrelated patterns of secondary structure and RBP binding throughout nuclear mRNAs that demarcate sites of alternative splicing and polyadenylation. We also uncover a collection of protein-bound sequence motifs, and identify their structural contexts, co-occurrences in transcripts encoding functionally related proteins, and interactions with putative RBPs. Finally, using these motifs, we find that the chloroplast RBP CP29A also interacts with nuclear mRNAs. In total, we provide a simultaneous view of the RNA secondary structure and RBP interaction landscapes in a eukaryotic nucleus.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Proteínas de Cloroplastos/metabolismo , RNA Mensageiro/metabolismo , RNA de Plantas/metabolismo , Ribonucleoproteínas/metabolismo , Arabidopsis/citologia , Arabidopsis/genética , Sequência de Bases , Sítios de Ligação , Sequência Consenso , Regulação da Expressão Gênica de Plantas , Conformação de Ácido Nucleico , Ligação Proteica , Transporte Proteico , Interferência de RNA , RNA Mensageiro/genética , RNA de Plantas/genética , Plântula/citologia , Plântula/genética , Plântula/metabolismo , Transcriptoma
16.
Int J Mol Sci ; 24(8)2023 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-37108518

RESUMO

The selectivity of drugs for G protein-coupled receptor (GPCR) signaling pathways is crucial for their therapeutic efficacy. Different agonists can cause receptors to recruit effector proteins at varying levels, thus inducing different signaling responses, called signaling bias. Although several GPCR-biased drugs are currently being developed, only a limited number of biased ligands have been identified regarding their signaling bias for the M1 muscarinic acetylcholine receptor (M1mAChR), and the mechanism is not yet well understood. In this study, we utilized bioluminescence resonance energy transfer (BRET) assays to compare the efficacy of six agonists in inducing Gαq and ß-arrestin2 binding to M1mAChR. Our findings reveal notable variations in agonist efficacy in the recruitment of Gαq and ß-arrestin2. Pilocarpine preferentially promoted the recruitment of ß-arrestin2 (∆∆RAi = -0.5), while McN-A-343 (∆∆RAi = 1.5), Xanomeline (∆∆RAi = 0.6), and Iperoxo (∆∆RAi = 0.3) exhibited a preference for the recruitment of Gαq. We also used commercial methods to verify the agonists and obtained consistent results. Molecular docking revealed that certain residues (e.g., Y404, located in TM7 of M1mAChR) could play crucial roles in Gαq signaling bias by interacting with McN-A-343, Xanomeline, and Iperoxo, whereas other residues (e.g., W378 and Y381, located in TM6) contributed to ß-arrestin recruitment by interacting with Pilocarpine. The preference of activated M1mAChR for different effectors may be due to significant conformational changes induced by biased agonists. By characterizing bias towards Gαq and ß-arrestin2 recruitment, our study provides insights into M1mAChR signaling bias.


Assuntos
Acetilcolina , Receptor Muscarínico M1 , Humanos , beta-Arrestinas/metabolismo , Simulação de Acoplamento Molecular , Receptor Muscarínico M1/metabolismo , Cloreto de (4-(m-Clorofenilcarbamoiloxi)-2-butinil)trimetilamônio , Pilocarpina/farmacologia , Proteínas de Ligação ao GTP/metabolismo , beta-Arrestina 2/metabolismo , beta-Arrestina 1/metabolismo , Transferência de Energia , Células HEK293
17.
Water Sci Technol ; 87(3): 798-811, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36789718

RESUMO

A calcined CaMgAl-layered double hydroxide nanocomposite, CaMgAl-LDH (CCMA-0.83-600), was prepared by introducing Mg on the basis of CaAl-LDHs for the removal of phosphate from wastewater. The structure of the as-synthesized CCMA-0.83-600 was confirmed by XRD and SEM analyses. Parameters affecting the adsorption process of phosphate adsorbed by CCMA-0.83-600 were thoroughly explored, such as initial pH, adsorbent dosage and co-existing anions. The adsorption kinetic study suggested that the adsorption process accorded with the pseudo-second-order kinetic model and the adsorption rate was controlled by both the liquid film diffusion and intra-particle diffusion. The adsorption isotherm study indicated the adsorption process followed by the Langmuir isotherm model. Thermodynamic analysis suggested that the adsorption of phosphate was spontaneous and exothermic. The obtained results indicated that CCMA-0.83-600 is a suitable candidate for the removal of phosphate from wastewater.


Assuntos
Fosfatos , Poluentes Químicos da Água , Águas Residuárias , Hidróxidos/química , Termodinâmica , Adsorção , Cinética , Poluentes Químicos da Água/química
18.
Appl Soft Comput ; 139: 110213, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37009545

RESUMO

The outbreak of Corona Virus Disease 2019 (COVID-19) makes people more concerned about the validity and timeliness of emergency decision making. When an emergency occurs, it is difficult for decision makers (DMs) to give accurate assessment information in the early stage due to the urgency of time, the incompleteness of information, and the limitations of DMs' cognition and knowledge. Hence, we use interval-valued intuitionistic hesitant fuzzy sets rather than exact numbers to better characterize the fuzziness and uncertainty of emergencies. In addition, the Internet has become a major platform for the public to express their opinions or concerns, so we can collect the user-generated content on social media to help DMs determine appropriate emergency decision-making criteria which are the premise and basis of scientific decisions. However, there is likely to be some correlation between the obtained criteria. To this end, we first extend the Bonferroni mean (BM) operator to the interval-valued intuitionistic hesitant fuzzy environment, and propose three interval-valued intuitionistic hesitant fuzzy BM operators to capture the interrelation of fuzzy input variables, including an interval-valued intuitionistic hesitant fuzzy BM operator, a simplified interval-valued intuitionistic hesitant fuzzy BM operator, and a simplified interval-valued intuitionistic hesitant fuzzy weighted BM (SIVIHFWBM) operator. Then, a new group emergency decision-making method based on the SIVIHFWBM operator and social media data is proposed, and the specific steps of ranking all emergency plans are put forward. Moreover, our method is applied to evaluate emergency plans for the prevention and control of COVID-19. Finally, the effectiveness and feasibility of the method are verified by the sensitivity analysis, validity test, and comparative analysis.

19.
Curr Issues Mol Biol ; 45(1): 311-326, 2022 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-36661508

RESUMO

(1) Background: C. vietnamensis is very suitable for growth in the low hilly areas of southern subtropical regions. Under appropriate conditions, the oil yield of C. vietnamensis can reach 1125 kg/ha (the existing varieties can reach 750 kg/ha). Moreover, the fruit of C. vietnamensis is large and the pericarp is thick (>5 cm). Therefore, a high seed ratio has become the main target economic trait for the breeding of C. vietnamensis. (2) Methods: A half-sibling population of C. vietnamensis plants with a combination of high and low seed ratios was constructed by crossing a C. vietnamensis female parent. Bulked segregant RNA analysis and full-length transcriptome sequencing were performed to determine the molecular mechanisms underlying a high seed ratio. (3) Results: Seed ratio is a complex quantitative trait with a normal distribution, which is significantly associated with four other traits of fruit (seed weight, seed number, fruit diameter, and pericarp thickness). Two candidate regions related to high seed ratio (HSR) were predicted. One spanned 140.8−148.4 Mb of chromosome 2 and was associated with 97 seed-yield-related candidate genes ranging in length from 278 to 16,628 bp. The other spanned 35.3−37.3 Mb on chromosome 15 and was associated with 38 genes ranging in length from 221 to 16,928 bp. Using the full-length transcript as a template, a total of 115 candidate transcripts were obtained, and 78 transcripts were predicted to be functionally annotated. The DEGs from two set pairs of cDNA sequencing bulks were enriched to cytochrome p450 CYP76F14 (KOG0156; GO:0055114, HSR4, HSR7), the gibberellin phytohormone pathway (GO:0016787, HSR5), the calcium signaling pathway (GO:0005509, HSR6), the polyubiquitin-PPAR signaling pathway (GO:0005515, HSR2, HSR3), and several main transcription factors (bZIP transcription factor, HSR1) in C. vietnamensis.

20.
Opt Lett ; 47(22): 5773-5776, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37219100

RESUMO

Phase measurement deflectometry (PMD) is an excellent method for high-precision optical surface measurement; through the simple system structure, accuracy comparable to that of interference methods can be realized. The key of PMD is to solve the ambiguity between the surface shape and normal vector. Considering all kinds of methods, the binocular PMD method is undoubtedly a method with a very simple system structure and can be easily applied to complex surfaces, such as free-form surfaces. However, this method relies on a large screen with high accuracy, which not only increases the weight of the system, but also reduces the flexibility of the system, and manufacturing errors in the large-size screen can easily become error sources. In this Letter, we have made some improvements based on the traditional binocular PMD. At first, we try to replace the large screen with two small screens to increase the flexibility and accuracy of the system. Further, we replace one small screen with a single point to simplify the system structure. Experiments show that the proposed methods not only improve the system flexibility and reduce the complexity, but can also achieve high measurement accuracy.

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