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1.
Plant Cell ; 36(7): 2550-2569, 2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38513608

RESUMEN

Embryo development in Arabidopsis (Arabidopsis thaliana) starts off with an asymmetric division of the zygote to generate the precursors of the embryo proper and the supporting extraembryonic suspensor. The suspensor degenerates as the development of the embryo proper proceeds beyond the heart stage. Until the globular stage, the suspensor maintains embryonic potential and can form embryos in the absence of the developing embryo proper. We report a mutant called meerling-1 (mrl-1), which shows a high penetrance of suspensor-derived polyembryony due to delayed development of the embryo proper. Eventually, embryos from both apical and suspensor lineages successfully develop into normal plants and complete their life cycle. We identified the causal mutation as a genomic rearrangement altering the promoter of the Arabidopsis U3 SMALL NUCLEOLAR RNA-ASSOCIATED PROTEIN 18 (UTP18) homolog that encodes a nucleolar-localized WD40-repeat protein involved in processing 18S preribosomal RNA. Accordingly, root-specific knockout of UTP18 caused growth arrest and accumulation of unprocessed 18S pre-rRNA. We generated the mrl-2 loss-of-function mutant and observed asynchronous megagametophyte development causing embryo sac abortion. Together, our results indicate that promoter rearrangement decreased UTP18 protein abundance during early stage embryo proper development, triggering suspensor-derived embryogenesis. Our data support the existence of noncell autonomous signaling from the embryo proper to prevent direct reprogramming of the suspensor toward embryonic fate.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Mutación , Semillas , Arabidopsis/genética , Arabidopsis/embriología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Mutación/genética , Semillas/genética , Semillas/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , ARN Ribosómico/genética
2.
Proc Natl Acad Sci U S A ; 120(29): e2301002120, 2023 07 18.
Artículo en Inglés | MEDLINE | ID: mdl-37428930

RESUMEN

Autophagy is a major means for the elimination of protein inclusions in neurons in neurodegenerative diseases such as Parkinson's disease (PD). Yet, the mechanism of autophagy in the other brain cell type, glia, is less well characterized and remains largely unknown. Here, we present evidence that the PD risk factor, Cyclin-G-associated kinase (GAK)/Drosophila homolog Auxilin (dAux), is a component in glial autophagy. The lack of GAK/dAux increases the autophagosome number and size in adult fly glia and mouse microglia, and generally up-regulates levels of components in the initiation and PI3K class III complexes. GAK/dAux interacts with the master initiation regulator UNC-51like autophagy activating kinase 1/Atg1 via its uncoating domain and regulates the trafficking of Atg1 and Atg9 to autophagosomes, hence controlling the onset of glial autophagy. On the other hand, lack of GAK/dAux impairs the autophagic flux and blocks substrate degradation, suggesting that GAK/dAux might play additional roles. Importantly, dAux contributes to PD-like symptoms including dopaminergic neurodegeneration and locomotor function in flies. Our findings identify an autophagy factor in glia; considering the pivotal role of glia under pathological conditions, targeting glial autophagy is potentially a therapeutic strategy for PD.


Asunto(s)
Proteínas de Drosophila , Enfermedad de Parkinson , Animales , Ratones , Drosophila/metabolismo , Auxilinas/metabolismo , Homólogo de la Proteína 1 Relacionada con la Autofagia/genética , Homólogo de la Proteína 1 Relacionada con la Autofagia/metabolismo , Autofagia , Ciclinas/metabolismo , Neuroglía/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Proteínas Relacionadas con la Autofagia/metabolismo , Proteínas de la Membrana/metabolismo
3.
Nucleic Acids Res ; 51(9): e52, 2023 05 22.
Artículo en Inglés | MEDLINE | ID: mdl-36971131

RESUMEN

A panel of unnatural base pairs is developed to expand genetic alphabets. One or more unnatural base pairs (UBPs) can be inserted to enlarge the capacity, diversity, and functionality of canonical DNA, so monitoring the multiple-UBPs-containing DNA by simple and convenient approaches is essential. Herein, we report a bridge-base approach to repurpose the capability of determining TPT3-NaM UBPs. The success of this approach depends on the design of isoTAT that can simultaneously pair with NaM and G as a bridge base, as well as the discovering of the transformation of NaM to A in absence of its complementary base. TPT3-NaM can be transferred to C-G or A-T by simple PCR assays with high read-through ratios and low sequence-dependent properties, permitting for the first time to dually locate the multiple sites of TPT3-NaM pairs. Then we show the unprecedented capacity of this approach to trace accurate changes and retention ratios of multiple TPT3-NaM UPBs during in vivo replications. In addition, the method can also be applied to identify multiple-site DNA lesions, transferring TPT3-NaM makers to different natural bases. Taken together, our work presents the first general and convenient approach capable of locating, tracing, and sequencing site- and number-unlimited TPT3-NaM pairs.


Asunto(s)
Emparejamiento Base , ADN , Emparejamiento Base/genética , ADN/análisis , ADN/química , ADN/genética , Replicación del ADN
4.
Traffic ; 23(10): 506-520, 2022 10.
Artículo en Inglés | MEDLINE | ID: mdl-36053864

RESUMEN

Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial and sporadic Parkinson's disease. A plethora of evidence has indicated a role for LRRK2 in endolysosomal trafficking in neurons, while LRRK2 function in glia, although highly expressed, remains largely unknown. Here, we present evidence that LRRK2/dLRRK mediates a lysosomal pathway that contributes to glial cell death and the survival of dopaminergic (DA) neurons. LRRK2/dLRRK knockdown in the immortalized microglia or flies results in enlarged and swelling lysosomes fewer in number. These lysosomes are less mobile, wrongly acidified, exhibit defective membrane permeability and reduced activity of the lysosome hydrolase cathepsin B. In addition, LRRK2/dLRRK depletion causes glial apoptosis, DA neurodegeneration, and locomotor deficits in an age-dependent manner. Taken together, these findings demonstrate a functional role of LRRK2/dLRRK in regulating the glial lysosomal pathway; deficits in lysosomal biogenesis and function linking to glial apoptosis potentially underlie the mechanism of DA neurodegeneration, providing insights on LRRK2/dLRRK function in normal and pathological brains.


Asunto(s)
Catepsina B , Neuronas Dopaminérgicas , Catepsina B/genética , Catepsina B/metabolismo , Muerte Celular , Neuronas Dopaminérgicas/metabolismo , Leucina/genética , Leucina/metabolismo , Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina/genética , Proteína 2 Quinasa Serina-Treonina Rica en Repeticiones de Leucina/metabolismo , Lisosomas/metabolismo , Mutación , Neuroglía/metabolismo
5.
BMC Bioinformatics ; 25(1): 32, 2024 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-38233745

RESUMEN

BACKGROUND: Epi-transcriptome regulation through post-transcriptional RNA modifications is essential for all RNA types. Precise recognition of RNA modifications is critical for understanding their functions and regulatory mechanisms. However, wet experimental methods are often costly and time-consuming, limiting their wide range of applications. Therefore, recent research has focused on developing computational methods, particularly deep learning (DL). Bidirectional long short-term memory (BiLSTM), convolutional neural network (CNN), and the transformer have demonstrated achievements in modification site prediction. However, BiLSTM cannot achieve parallel computation, leading to a long training time, CNN cannot learn the dependencies of the long distance of the sequence, and the Transformer lacks information interaction with sequences at different scales. This insight underscores the necessity for continued research and development in natural language processing (NLP) and DL to devise an enhanced prediction framework that can effectively address the challenges presented. RESULTS: This study presents a multi-scale self- and cross-attention network (MSCAN) to identify the RNA methylation site using an NLP and DL way. Experiment results on twelve RNA modification sites (m6A, m1A, m5C, m5U, m6Am, m7G, Ψ, I, Am, Cm, Gm, and Um) reveal that the area under the receiver operating characteristic of MSCAN obtains respectively 98.34%, 85.41%, 97.29%, 96.74%, 99.04%, 79.94%, 76.22%, 65.69%, 92.92%, 92.03%, 95.77%, 89.66%, which is better than the state-of-the-art prediction model. This indicates that the model has strong generalization capabilities. Furthermore, MSCAN reveals a strong association among different types of RNA modifications from an experimental perspective. A user-friendly web server for predicting twelve widely occurring human RNA modification sites (m6A, m1A, m5C, m5U, m6Am, m7G, Ψ, I, Am, Cm, Gm, and Um) is available at http://47.242.23.141/MSCAN/index.php . CONCLUSIONS: A predictor framework has been developed through binary classification to predict RNA methylation sites.


Asunto(s)
Metilación de ARN , ARN , Humanos , ARN/genética , Redes Neurales de la Computación , Metilación , Procesamiento Postranscripcional del ARN
6.
BMC Cancer ; 24(1): 26, 2024 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-38166756

RESUMEN

BACKGROUND: Epigenetic alterations contribute greatly to the development and progression of colorectal cancer, and effect of aberrant miR-622 expression is still controversial. This study aimed to discover miR-622 regulation in CRC proliferation. METHODS: miR-622 expression and prognosis were analyzed in clinical CRC samples from Nanfang Hospital. miR-622 regulation on cell cycle and tumor proliferation was discovered, and FOLR2 was screened as functional target of miR-622 using bioinformatics analysis, which was validated via dual luciferase assay and gain-of-function and loss-of-function experiments both in vitro and in vivo. RESULTS: miR-622 overexpression in CRC indicated unfavorable prognosis and it regulated cell cycle to promote tumor growth both in vitro and in vivo. FOLR2 is a specific, functional target of miR-622, which negatively correlates with signature genes in cell cycle process to promote CRC proliferation. CONCLUSIONS: miR-622 upregulates cell cycle process by targeting FOLR2 to promote CRC proliferation, proposing a novel mechanism and treatment target in CRC epigenetic regulation of miR-622.


Asunto(s)
Proliferación Celular , Neoplasias Colorrectales , Receptor 2 de Folato , MicroARNs , Humanos , Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Epigénesis Genética , Receptor 2 de Folato/genética , Receptor 2 de Folato/metabolismo , Regulación Neoplásica de la Expresión Génica , MicroARNs/metabolismo
7.
Microb Cell Fact ; 23(1): 153, 2024 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-38796416

RESUMEN

BACKGROUND: Dihydroxyacetone (DHA) stands as a crucial chemical material extensively utilized in the cosmetics industry. DHA production through the dephosphorylation of dihydroxyacetone phosphate, an intermediate product of the glycolysis pathway in Escherichia coli, presents a prospective alternative for industrial production. However, insights into the pivotal enzyme, dihydroxyacetone phosphate dephosphorylase (HdpA), remain limited for informed engineering. Consequently, the development of an efficient tool for high-throughput screening of HdpA hypermutants becomes imperative. RESULTS: This study introduces a methylglyoxal biosensor, based on the formaldehyde-responding regulator FrmR, for the selection of HdpA. Initial modifications involved the insertion of the FrmR binding site upstream of the -35 region and into the spacer region between the -10 and -35 regions of the constitutive promoter J23110. Although the hybrid promoter retained constitutive expression, expression of FrmR led to complete repression. The addition of 350 µM methylglyoxal promptly alleviated FrmR inhibition, enhancing promoter activity by more than 40-fold. The methylglyoxal biosensor system exhibited a gradual increase in fluorescence intensity with methylglyoxal concentrations ranging from 10 to 500 µM. Notably, the biosensor system responded to methylglyoxal spontaneously converted from added DHA, facilitating the separation of DHA producing and non-producing strains through flow cytometry sorting. Subsequently, the methylglyoxal biosensor was successfully applied to screen a library of HdpA mutants, identifying two strains harboring specific mutants 267G > T and D110G/G151C that showed improved DHA production by 68% and 114%, respectively. Expressing of these two HdpA mutants directly in a DHA-producing strain also increased DHA production from 1.45 to 1.92 and 2.29 g/L, respectively, demonstrating the enhanced enzyme properties of the HdpA mutants. CONCLUSIONS: The methylglyoxal biosensor offers a novel strategy for constructing genetically encoded biosensors and serves as a robust platform for indirectly determining DHA levels by responding to methylglyoxal. This property enables efficiently screening of HdpA hypermutants to enhance DHA production.


Asunto(s)
Técnicas Biosensibles , Dihidroxiacetona , Escherichia coli , Piruvaldehído , Piruvaldehído/metabolismo , Técnicas Biosensibles/métodos , Dihidroxiacetona/metabolismo , Escherichia coli/metabolismo , Escherichia coli/genética , Regiones Promotoras Genéticas , Ingeniería Metabólica/métodos , Proteínas de Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética
8.
Proc Natl Acad Sci U S A ; 118(23)2021 06 08.
Artículo en Inglés | MEDLINE | ID: mdl-34078666

RESUMEN

Intrinsic mechanisms such as temporal series of transcription factors orchestrate neurogenesis from a limited number of neural progenitors in the brain. Extrinsic regulations, however, remain largely unexplored. Here we describe a two-step glia-derived signal that regulates neurogenesis in the Drosophila mushroom body (MB). In a temporal manner, glial-specific ubiquitin ligase dSmurf activates non-cell-autonomous Hedgehog signaling propagation by targeting the receptor Patched to suppress and promote the exit of MB neuroblast (NB) proliferation, thereby specifying the correct α/ß cell number without affecting differentiation. Independent of NB proliferation, dSmurf also stabilizes the expression of the cell-adhesion molecule Fasciclin II (FasII) via its WW domains and regulates FasII homophilic interaction between glia and MB axons to refine α/ß-lobe integrity. Our findings provide insights into how extrinsic glia-to-neuron communication coordinates with NB proliferation capacity to regulate MB neurogenesis; glial proteostasis is likely a generalized mechanism in orchestrating neurogenesis.


Asunto(s)
Comunicación Celular , Proliferación Celular , Cuerpos Pedunculados/embriología , Neurogénesis , Neuroglía/metabolismo , Animales , Proteínas de Drosophila/metabolismo , Drosophila melanogaster
9.
J Am Chem Soc ; 145(40): 22079-22085, 2023 Oct 11.
Artículo en Inglés | MEDLINE | ID: mdl-37784238

RESUMEN

Due to the enormous chemical and structural diversities and designable properties and functionalities, covalent organic frameworks (COFs) hold great promise as tailored materials for industrial applications in electronics, biology, and energy technologies. They were typically obtained as partially crystalline materials, although a few single-crystal three-dimensional (3D) COFs have been obtained recently with structures probed by diffraction techniques. However, it remains challenging to grow single-crystal COFs with controlled morphology and to elucidate the local structures of 3D COFs, imposing severe limitations on the applications and understanding of the local structure-property correlations. Herein, we develop a method for designed growth of five types of single crystalline flakes of 3D COFs with controlled morphology, front crystal facets, and defined edge structures as well as surface chemistry using surfactants that can be self-assembled into layered structures to confine crystal growth in water. The flakes enable direct observation of local structures including monomer units, pore structure, edge structure, grain boundary, and lattice distortion of 3D COFs as well as gradually curved surfaces in kinked but single crystalline 3D COFs with a resolution of up to ∼1.7 Å. In comparison with flakes of two-dimensional crystals, the synthesized flakes show much higher chemical, mechanical, and thermal stability.

10.
Anal Chem ; 95(44): 16210-16215, 2023 11 07.
Artículo en Inglés | MEDLINE | ID: mdl-37899593

RESUMEN

Tuberculosis (TB) is a chronic systemic infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). Methionine aminopeptidase 1 (MtMET-AP1) is a hydrolase that mediates the necessary post-translational N-terminal methionine excision (NME) of peptides during protein synthesis, which is necessary for bacterial proliferation and is a potential target for the treatment of tuberculosis. Based on the functional characteristics of MtMET-AP1, we developed an enzymatic activated near-infrared fluorescent probe DDAN-MT for rapid, highly selective, and real-time monitoring of endogenous MtMET-AP1 activity in M. tuberculosis. Using the probe DDAN-MT, a visually high-throughput screening technique was established, which obtained three potential inhibitors (GSK-J4 hydrochchloride, JX06, and lavendustin C) against MtMET-AP1 from a 2560 compounds library. More importantly, these inhibitors could inhibit the growth of M. tuberculosis H37Ra especially (MICs < 5 µM), with low toxicities on intestinal bacteria strains and human cells. Therefore, the visual sensing of MtMET-AP1 was successfully performed by DDAN-MT, and MtMET-AP1 inhibitors were discovered as potential antituberculosis agents.


Asunto(s)
Mycobacterium tuberculosis , Tuberculosis , Humanos , Antituberculosos/farmacología , Mycobacterium tuberculosis/metabolismo , Colorantes Fluorescentes , Pruebas de Sensibilidad Microbiana , Aminopeptidasas/metabolismo
11.
Plant Cell ; 32(2): 392-413, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31806675

RESUMEN

The spikelet is an inflorescence structure unique to grasses. The molecular mechanisms underlying spikelet development and evolution are unclear. In this study, we characterized three allelic recessive mutants in rice (Oryza sativa): nonstop glumes 1-1 (nsg1-1), nsg1-2, and nsg1-3 In these mutants, organs such as the rudimentary glume, sterile lemma, palea, lodicule, and filament were elongated and/or widened, or transformed into lemma- and/or marginal region of the palea-like organs. NSG1 encoded a member of the C2H2 zinc finger protein family and was expressed mainly in the organ primordia of the spikelet. In the nsg1-1 mutant spikelet, LHS1 DL, and MFO1 were ectopically expressed in two or more organs, including the rudimentary glume, sterile lemma, palea, lodicule, and stamen, whereas G1 was downregulated in the rudimentary glume and sterile lemma. Furthermore, the NSG1 protein was able to bind to regulatory regions of LHS1 and then recruit the corepressor TOPLESS-RELATED PROTEIN to repress expression by downregulating histone acetylation levels of the chromatin. The results suggest that NSG1 plays a pivotal role in maintaining organ identities in the spikelet by repressing the expression of LHS1, DL, and MFO1.


Asunto(s)
Dedos de Zinc CYS2-HIS2/genética , Oryza/genética , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/genética , Ingeniería Genética , Inflorescencia , Mutación , Fenotipo , Transcriptoma
12.
Chemistry ; 29(10): e202203108, 2023 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-36401597

RESUMEN

Due to the limited resources and high cost of noble metals, boosting their catalytic activities is highly desired in the current catalysis industry. Here, we report a synergetic catalyst, combining Pd2+ and Pd0 species in a nitrogen-doped porous carbons (NPC), which shows boosted catalytic activities in hydrogenation reactions of organic nitro compounds (nitrobenzene, 4-nitrophenol, 1-nitronaphthalene and 1-nitropropane) under near ambient conditions. This synergetic catalyst NPC-[Pd] was synthesized by partial reduction of a palladium-loaded NPC. The catalytic activities and selectivity of NPC-[Pd] for hydrogenation were enhanced significantly compared with those of NPC-Pd2+ or NPC-Pd0 nanoparticles. Theoretical calculations show that H2 preferentially dissociates on Pd nanoparticles, and then organic molecules (nitrobenzene) can be captured and react with the dissociated H atom on Pd2+ sites. Similar reaction procedure also occur on Pt or Rh. Hydrogenation of different aromatic compounds with different functional groups (naphthalene, 4-nitrochlorobenzene, benzaldehyde and acetophenone) confirmed the broad excellent catalytic activity of this synergistic catalyst.

13.
Ann Hematol ; 102(2): 299-309, 2023 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-36607351

RESUMEN

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonal disease of hematopoietic stem cells (HSCs). Long noncoding RNAs (lncRNAs) perform a wide range of biological functions, including the regulation of gene expression, cell differentiation, and proliferation, but their role in PNH remains unclear.CD59- and CD59+ granulocytes and monocytes from 35 PNH patients were sorted. High-throughput sequencing was analyzed in 5 PNH patients, and differentially expressed lncRNAs and mRNAs were identified. The mRNAs with fragments per kilobase of exon model per million mapped fragments (FPKM) > 10 in at least 3 patients were selected, and experiments were performed to identify their upstream regulatory lncRNAs. The expression of selected mRNAs and lncRNAs was verified by qRT‒PCR, and the correlation of these expression patterns with clinical data from other 30 PNH patients was analyzed. Then, the functions of the lncRNAs were studied in the PIGA-KO-THP-1 cell line.Transcription analysis revealed 742 upregulated and 1376 downregulated lncRNAs and 3276 upregulated and 213 downregulated mRNAs. After deep screening, 8 highly expressed mRNAs that were related to the NF-κB pathway were analyzed to determine coexpression patterns. LINC01002, FAM157C, CTD-2530H12.2, XLOC-064331 and XLOC-106677 were correlated with the 8 mRNAs. After measuring the expression of these molecules in 30 PNH patients by qRT‒PCR, lncRNA FAM157C was verified to be upregulated in the PNH clone, and its expression levels were positively correlated with the LDH levels and CD59- granulated and monocyte cell ratios. After knockdown of the FAM157C gene in the PIGA-KO-THP-1 cell line, we found that the cells were arrested in the G0/G1 phase and S phase, the apoptosis rate increased, and the cell proliferation decreased.LncRNA FAM157C was proven to promote PNH clone proliferation, and this is the first study to explore the role of lncRNAs in PNH.


Asunto(s)
Hemoglobinuria Paroxística , ARN Largo no Codificante , Humanos , Hemoglobinuria Paroxística/genética , Hemoglobinuria Paroxística/diagnóstico , ARN Largo no Codificante/genética , Células Madre Hematopoyéticas/metabolismo , Células Clonales/química , Antígenos CD59/análisis , Antígenos CD59/metabolismo , Proliferación Celular/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo
14.
Bioorg Chem ; 140: 106827, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37683537

RESUMEN

The high fidelity poses a central role in developing unnatural base pairs (UBPs), which means the high pairing capacity of unnatural bases with their partners, and low mispairing with all the natural bases. Different strategies have been used to develop higher-fidelity UBPs, including optimizing hydrophobic interaction forces between UBPs. Variant substituent groups are allowed to fine tune the hydrophobic forces of different UBPs' candidates. However, the modifications on the skeleton of TPT3 base are rare and the replication fidelity of TPT3-NaM remains hardly to improve so far. In this paper, we reasoned that modifying and/or expanding the aromatic surface by Bromo-substituents to slightly increase hydrophobicity of TPT3 might offer a way to increase the fidelity of this pair. Based on the hypothesis, we synthesized the bromine substituted TPT3, 2-bromo-TPT3 and 2, 4-dibromo-TPT3 as the new TPT3 analogs. While the enzyme reaction kinetic experiments showed that d2-bromo-TPT3-dNaM pair and d2, 4-dibromo-TPT3TP-dNaM pair had slightly less efficient incorporation and extension rates than that of dTPT3-dNaM pair, the assays did reveal that the mispairing of 2-bromo-TPT3 and 2, 4-dibromo-TPT3 with all the natural bases could dramatically decrease in contrast to TPT3. Their lower mispairing capacity promoted us to run polymerase chain amplification reactions, and a higher fidelity of d2-bromo-TPT3-dNaM pair could be obtained with 99.72 ± 0.01% of the in vitro replication fidelity than that of dTPT3-dNaM pair, 99.52 ± 0.09%. In addition, d2-bromo-TPT3-dNaM can also be effectively copied in E. coli cells, which showed the same replication fidelity as that of dTPT3-dNaM in the specific sequence, but a higher fidelity in the random sequence context.


Asunto(s)
Emparejamiento Base , Bromo , Replicación del ADN , Humanos , Escherichia coli , Cinética
15.
BMC Vet Res ; 19(1): 19, 2023 Jan 21.
Artículo en Inglés | MEDLINE | ID: mdl-36681807

RESUMEN

Escherichia coli (E. coli) is an opportunistic pathogen that can cause clinical mastitis in dairy cows worldwide. Mastitis produces severe symptoms in dairy cows, such as udder inflammation, the production of harmful substances, reduced milk production, and altered milk quality. Intramammary injections of rifaximin have a beneficial effect on dairy cow mastitis, especially for mastitis caused by E. coli. However, we do not know whether the currently accepted clinical administration scheme is reasonable. Therefore, the purpose of this experiment was to evaluate the clinical dosing regimen for curing mastitis induced by E. coli. In this study, the pharmacokinetics of four single dose groups (50, 100, 200, and 400 µg/gland) were studied in CD-1 lactating mice, and the main pharmacokinetic parameters were obtained by non-compartment and two-compartment model of Phoenix 8.1 software. A total of 5,000 colony-forming units (CFU) of E. coli ATCC25922 were injected into the mammary glands of mice under anatomic microscope guidance. After 12 h of growth in vivo, the mouse mastitis model was successfully developed. In pharmacodynamics experiment, 12 different dosing regimens (doses ranged from 25 to 800 µg/gland and two dosing intervals of 12 and 24 h) were used to study the therapeutic potential of rifaximin for mastitis. The PK/PD model was established by integrating pharmacokinetics and pharmacodynamics using the inhibitory sigmoid Emax model. The optimal antibacterial effect was 2log10CFU/gland reduction of bacterial colony counts in vivo, when the magnitude of AUC24/MIC exceeded 57.80 h. A total of 57.80 h of AUC24/MIC was defined as a target value in the Monte Carlo simulation. The clinically recommended dosage regimen of 100 mg/gland every 12 h in a day achieved a 91.08% cure rate for the treatment of bovine mastitis caused by E. coli infection.


Asunto(s)
Enfermedades de los Bovinos , Infecciones por Escherichia coli , Mastitis Bovina , Femenino , Bovinos , Animales , Ratones , Escherichia coli , Rifaximina/uso terapéutico , Lactancia , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/veterinaria , Leche/microbiología , Mastitis Bovina/tratamiento farmacológico , Mastitis Bovina/microbiología , Glándulas Mamarias Animales
16.
BMC Biol ; 20(1): 34, 2022 02 07.
Artículo en Inglés | MEDLINE | ID: mdl-35130883

RESUMEN

BACKGROUND: In insects, airborne chemical signals are mainly detected by two receptor families, odorant receptors (ORs) and ionotropic receptors (IRs). Functions of ORs have been intensively investigated in Diptera and Lepidoptera, while the functions and evolution of the more ancient IR family remain largely unexplored beyond Diptera. RESULTS: Here, we identified a repertoire of 26 IRs from transcriptomes of female and male antennae, and ovipositors in the moth Agrotis segetum. We observed that a large clade formed by IR75p and IR75q expansions is closely related to the acid-sensing IRs identified in Diptera. We functionally assayed each of the five AsegIRs from this clade using Xenopus oocytes and found that two receptors responded to the tested ligands. AsegIR75p.1 responded to several compounds but hexanoic acid was revealed to be the primary ligand, and AsegIR75q.1 responded primarily to octanoic acid, and less so to nonanoic acid. It has been reported that the C6-C10 medium-chain fatty acids repel various insects including many drosophilids and mosquitos. We show that the C6-C10 medium-chain fatty acids elicited antennal responses of both sexes of A. segetum, while only octanoic acid had repellent effect to the moths in a behavioral assay. In addition, using fluorescence in situ hybridization, we demonstrated that the five IRs and their co-receptor AsegIR8a are not located in coeloconic sensilla as found in Drosophila, but in basiconic or trichoid sensilla. CONCLUSIONS: Our results significantly expand the current knowledge of the insect IR family. Based on the functional data in combination with phylogenetic analysis, we propose that subfunctionalization after gene duplication plays an important role in the evolution of ligand specificities of the acid-sensing IRs in Lepidoptera.


Asunto(s)
Brassica napus , Dípteros , Mariposas Nocturnas , Receptores Odorantes , Animales , Antenas de Artrópodos , Caprilatos , Dípteros/genética , Femenino , Hibridación Fluorescente in Situ , Proteínas de Insectos/genética , Ligandos , Masculino , Mariposas Nocturnas/genética , Filogenia , Receptores Odorantes/genética
17.
BMC Biol ; 20(1): 80, 2022 03 31.
Artículo en Inglés | MEDLINE | ID: mdl-35361182

RESUMEN

BACKGROUND: Using genetically modified plants as natural dispensers of insect pheromones may eventually become part of a novel strategy for integrated pest management. RESULTS: In the present study, we first characterized essential functional genes for sex pheromone biosynthesis in the rice stem borer Chilo suppressalis (Walker) by heterologous expression in Saccharomyces cerevisiae and Nicotiana benthamiana, including two desaturase genes CsupYPAQ and CsupKPSE and a reductase gene CsupFAR2. Subsequently, we co-expressed CsupYPAQ and CsupFAR2 together with the previously characterized moth desaturase Atr∆11 in N. benthamiana. This resulted in the production of (Z)-11-hexadecenol together with (Z)-11-hexadecenal, the major pheromone component of C. suppressalis. Both compounds were collected from the transformed N. benthamiana headspace volatiles using solid-phase microextraction. We finally added the expression of a yeast acetyltransferase gene ATF1 and could then confirm also (Z)-11-hexadecenyl acetate release from the plant. CONCLUSIONS: Our results pave the way for stable transformation of plants to be used as biological pheromone sources in different pest control strategies.


Asunto(s)
Mariposas Nocturnas , Atractivos Sexuales , Animales , Mariposas Nocturnas/genética , Feromonas/metabolismo , Nicotiana/genética
18.
Sensors (Basel) ; 23(6)2023 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-36991592

RESUMEN

In order to accurately predict the gas concentration, find out the gas abnormal emission in advance, and take effective measures to reduce the gas concentration in time, this paper analyzes multivariate monitoring data and proposes a new dynamic combined prediction method of gas concentration. Spearman's rank correlation coefficient is applied for the dynamic optimization of prediction indicators. The time series and spatial topology features of the optimized indicators are extracted and input into the combined prediction model of gas concentration based on indicators dynamic optimization and Bi-LSTMs (Bi-directional Long Short-term Memory), which can predict the gas concentration for the next 30 min. The results show that the other gas concentration, temperature, and humidity indicators are strongly correlated with the gas concentration to be predicted, and Spearman's rank correlation coefficient is up to 0.92 at most. The average R2 of predicted value and real value is 0.965, and the average prediction efficiency R for gas abnormal or normal emission is 79.9%. Compared with the other models, the proposed dynamic optimized indicators combined model is more accurate, and the missing alarm of gas abnormal emission is significantly alleviated, which greatly improves the early alarming accuracy. It can assist the safety monitoring personnel in decision making and has certain significance to improve the safety production efficiency of coal mines.

19.
J Environ Manage ; 336: 117624, 2023 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-36868152

RESUMEN

To mitigate aviation's carbon emissions of the aviation industry, the following steps are vital: accurately quantifying the carbon emission path by considering uncertainty factors, including transportation demand in the post-COVID-19 pandemic period; identifying gaps between this path and emission reduction targets; and providing mitigation measures. Some mitigation measures that can be employed by China's civil aviation industry include the gradual realization of large-scale production of sustainable aviation fuels and transition to 100% sustainable and low-carbon sources of energy. This study identified the key driving factors of carbon emissions by using the Delphi Method and set scenarios that consider uncertainty, such as aviation development and emission reduction policies. A backpropagation neural network and Monte Carlo simulation were used to quantify the carbon emission path. The study results show that China's civil aviation industry can effectively help the country achieve its carbon peak and carbon neutrality goals. However, to achieve the net-zero carbon emissions goal of global aviation, China needs to reduce its emissions by approximately 82%-91% based on the optimal emission scenario. Thus, under the international net-zero target, China's civil aviation industry will face significant pressure to reduce its emissions. The use of sustainable aviation fuels is the best way to reduce aviation emissions by 2050. Moreover, in addition to the application of sustainable aviation fuel, it will be necessary to develop a new generation of aircraft introducing new materials and upgrading technology, implement additional carbon absorption measures, and make use of carbon trading markets to facilitate China's civil aviation industry's contribution to reduce climate change.


Asunto(s)
Aviación , COVID-19 , Humanos , Dióxido de Carbono/análisis , Incertidumbre , Pandemias , COVID-19/prevención & control , Desarrollo Económico , China , Carbono/análisis
20.
Angew Chem Int Ed Engl ; 62(17): e202217253, 2023 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-36744542

RESUMEN

Two-dimensional (2D) materials catalysts provide an atomic-scale view on a fascinating arena for understanding the mechanism of electrocatalytic carbon dioxide reduction (CO2 ECR). Here, we successfully exfoliated both layered and nonlayered ultra-thin metal phosphorous trichalcogenides (MPCh3 ) nanosheets via wet grinding exfoliation (WGE), and systematically investigated the mechanism of MPCh3 as catalysts for CO2 ECR. Unlike the layered CoPS3 and NiPS3 nanosheets, the active Sn atoms tend to be exposed on the surfaces of nonlayered SnPS3 nanosheets. Correspondingly, the nonlayered SnPS3 nanosheets exhibit clearly improved catalytic activity, showing formic acid selectivity up to 31.6 % with -7.51 mA cm-2 at -0.65 V vs. RHE. The enhanced catalytic performance can be attributed to the formation of HCOO* via the first proton-electron pair addition on the SnPS3 surface. These results provide a new avenue to understand the novel CO2 ECR mechanism of Sn-based and MPCh3 -based catalysts.

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